S.N. Elansky, L.Yu. Kokaeva, N.V. Statsyuk, Yu.T. Dyakov
Isingeniso
I-Oomycete Phytophthora infestans (Mont.) I-De Bary - i-agent ebangela ukubola okuphuzile, isifo esibaluleke kakhulu kwezomnotho samazambane notamatisi - idonse ukunaka kwabacwaningi abavela emazweni ahlukene iminyaka engaphezu kwekhulu nesigamu. Ngokungazelelwe ebonakala eYurophu phakathi nekhulu le-XNUMX, kwabangela ubhubhane lwamazambane oluye lwahlala enkumbulweni yezizukulwane eziningi.
Kuze kube manje, kuvame ukubizwa ngokuthi "amakhowe wendlala yase-Ireland". Cishe iminyaka eyikhulu ngemuva kwezifo eziwumshayabhuqe zokuqala, kwatholakala izinhlobo zamazambane asendle aseMexico amelana nokubola sekwephuzile, izindlela zokuwawela ngamazambane atshaliwe (Muller, 1935), futhi kwatholakala izinhlobo zokuqala ezimelana nokubola (Pushkarev, 1937) . Kodwa-ke, ngemva nje kokuqala kokulinywa kwabo kwezentengiselwano, kwanqwabelana izinhlanga ze-pathogen ye-blight sekwephuzile, eyingozi ezinhlobonhlobo ezimelana nezifo. kanye nezakhi zofuzo ezintsha zokumelana ezethulwa ezinhlobonhlobo ezivela kumazambane asendle aseMexico zaqala ukulahlekelwa ukusebenza kwazo ngokushesha.
Ukwehluleka ukusetshenziswa kokumelana ne-monogenic (okuqondile) kuphoqe abalimi ukuthi babheke izindlela eziyinkimbinkimbi zokusebenzisa ukumelana ne-polygenic (okuvundlile) okungaqondile. Eminyakeni yamuva nje, kubantu ngabanye be-parasite, izinhlanga ezinolaka kakhulu seziqalile ukunqwabelana, okubangela ukuguguleka kokumelana nokungaqondile. Ukufika kwezinhlobo ezimelana nesikhunta sekudale izinkinga ekusebenziseni amakhemikhali okuvikela amazambane.
Ngenxa yomehluko omkhulu phakathi kwe-oomycetes nesikhunta ekwakhiweni kwamakhemikhali, i-ultrastructure kanye nemetabolism, ama-fungicides, ikakhulukazi lawo ahlelekile asetshenziselwa ukuvikela izitshalo ezifweni eziningi zesikhunta, awasebenzi ekulweni ne-oomycetes.
Ngakho-ke, ekuvikelweni kwamakhemikhali ngokumelene nokulimaza sekwephuzile, kusetshenziswe izikhathi eziningi (kufika ku-12 ngesizini ngayinye noma ngaphezulu) ngamalungiselelo okuxhumana ochungechunge olubanzi lwesenzo. Isinyathelo soguquko kwaba ukusetshenziswa kwama-phenylamide, anobuthi kuma-oomycetes futhi asakazeka ngendlela ehlelekile ezitshalweni. Kodwa-ke, ukusetshenziswa kwabo okubanzi ngokushesha kwaholela ekuqoqweni kwezinhlobo ezimelana nesikhunta (Davidse et al., 1981), okwaba nzima kakhulu ukuvikela izitshalo. I-P. infestans iyona kuphela i-parasite endaweni epholile, umonakalo lapho ukulima okuphilayo ngeke kuncishiswe ngaphandle kokusebenzisa amakhemikhali okuzivikela (Van Bruggen, 1995).
Okungenhla kuchaza ukunakwa okukhulu okukhokhwa abacwaningi abavela emazweni ahlukene ocwaningweni lwabantu be-P. infestans, ukuguquguquka kobuningi babo kanye nokwakheka kwezakhi zofuzo, kanye nezindlela zofuzo zokuhlukahluka.
Umjikelezo wokuphila we-R. INFESTANS
I-Oomycete Phytophthora infestans yakha i-mycelium ye-intercellular ene-haustoria ngaphakathi kwamaqabunga amazambane. Ukondla izicubu zamaqabunga, kubangela ukwakheka kwamabala amnyama, aphenduke abe mnyama futhi abole esimweni sezulu esimanzi. Ngokuhlulwa okukhulu, lonke iqabunga liyafa. Ngemuva kwesikhathi sokudla, ama-outgrows akhiwa ku-mycelium - sporangiophores - ekhula ngaphandle ngokusebenzisa i-stomata. Esimeni sezulu esimanzi, benza ukuqhakaza okumhlophe ezungeze amabala angaphansi kwamaqabunga. Emaphethelweni e-sporangiophores, i-zoosporangia emise okwe-lemon iyakhiwa, ephuka futhi ithwalwe yi-spray of rain (Fig. 1). Ukungena emathonsini amanzi phezu kweqabunga lamazambane, i-sporangia iqhuma ngama-zoospores angu-6-8, okuthi, ngemva kwesikhathi sokunyakaza, ayindilinga, embozwe ngegobolondo futhi ahlume ngeshubhu elihluma. Ihlumela lingena nge-stomata lingene esicupheni seqabunga. Ngaphansi kwezimo ezithile, i-sporangia ingakhula njengeshubhu lokukhula ngokuqondile libe yizicubu zamaqabunga. Ngaphansi kwezimo ezivumayo, isikhathi kusukela ekuthelelekeni kuya ekwakhiweni kwe-sporulation entsha yizinsuku ezingu-3-4 kuphela.
Uma iphansi futhi ihlungiwe enhlabathini, i-sporangia iyakwazi ukuthelela izilimo eziyizigaxa. Izilimo eziyizigaxa ezithinteke kakhulu ziyabola ngesikhathi sokugcinwa; kwabathintekile ababuthakathaka, ukutheleleka kungase kuqhubeke kuze kube isizini elandelayo. Ngaphezu kwalokho, i-causative agent ye-blight sekwephuzile ingaqhubeka ebusika ngendlela yama-oospores (izinhlamvu zocansi eziminyene ezinodonga oluqinile) enhlabathini emfucumfucu yezitshalo nasembewu katamatisi. Ama-oospores akhiwa ezithweni zezitshalo eziphilayo lapho izinhlobo zezinhlobo ezahlukene zokukhwelana zihlangana nomswakama owedlulele. Entwasahlobo, i-asexual sporulation yakheka ezigabeni ezitshalwe ezinegciwane futhi ezinsalela zezitshalo ezinama-oospores, ama-zoospores angena enhlabathini futhi abangele ukutheleleka kwamaqabunga aphansi ezitshalo. Kwezinye izimo, i-mycelium ingakhula kusuka ku-tuber enegciwane eduze kwengxenye eluhlaza yesitshalo futhi ngokuvamile ivela engxenyeni engenhla yesiqu.
Umehluko obalulekile phakathi kwama-oomycetes kanye nesikhunta esiningi usekuveleni kwe-diplophase emjikelezweni wabo wempilo nge-gametic meiosis kanye nokuhluma kwama-zygotes (oospores) ngaphandle kwe-reductive nuclear fission. Lesi sici, kanye ne-dipolar heterotallism ethatha isikhundla sobulili obubili, kubonakala sengathi senza kube nokwenzeka ukusebenzisa kuma-oomycetes izindlela ezithuthukisiwe zokutadisha isibalo sama-eukaryote aphezulu (ukuhlaziywa kwe-panmixia nokuhlukaniswa iziqephu kwezibalo zabantu, ukugeleza kwezakhi zofuzo zangaphakathi kanye ne-interpopulation, njll.). Kodwa-ke, izici ezintathu azikuvumeli ukudlulisa ngokuphelele lezi zindlela ocwaningweni lwabantu be-P. infestans.
1. Kanye nama-oospores ayi-hybrid, ama-oospores azivundile kanye ne-parthenogenetic akhiwa ngenani labantu (uFife noShaw, 1992; u-Anikina et al., 1997a; Savenkova, Cherepnikoba-Anirina, 2002; Smirnov, 2003), kanye nemvamisa yokwakheka kwawo. ingase ibe nethonya emiphumeleni yokuhlolwa.
2. Inqubo yocansi ku-P. infestans yenza umnikelo ongabalulekile ekuguquguqukeni kobukhulu besibalo sabantu, ngoba isikhunta sikhiqiza ikakhulukazi ngezinhlamvu zemifino, zakha ngaphezu kwe-90% yemiphumela yokuhlaziywa kohlobo lokukhwelana ngendlela yendabuko. endaweni yezakhamzimba... inkathi yokukhula izizukulwane ezimbalwa ze-asexual sporulation (ukuthuthukiswa kwesifo se-polycyclic). Ama-Oospores adlala indima ebalulekile ekulondolozweni kwento ephilayo phakathi nenkathi lapho zingekho izitshalo eziluhlaza (ebusika) kanye nasekuthelelekeni okuyinhloko kwezithombo. Khona-ke, ngesikhathi sasehlobo, ukukhiqizwa kwe-clonal kwenzeka futhi ukwanda noma, ngokuphambene, ukwehla kwenani lama-clones ngabanye avela ngenxa yokuhlanganiswa kocansi, okunqunywa ngokuyinhloko ukukhethwa kokuguquguquka okwengeziwe. Ngakho-ke, isilinganiso sama-clones angawodwana esibalweni sabantu ekuqaleni nasekupheleni kwe-epiphytotics singahluka ngokuphelele.
3. Umjikelezo ochaziwe ujwayelekile kubantu bomdabu be-P. infestans ezweni labo, eMelika Ephakathi. Kwezinye izindawo zomhlaba, inqubo yocansi yayingaziwa iminyaka engaphezu kweyi-100; i-vegetative mycelium ezigabeni zamazambane ezinegciwane kwakuyisigaba sobusika. Umjikelezo wokuphila wawune-agamic ngokuphelele, futhi ukusabalala kwakugxile emvelweni: ukutheleleka okuvela ezigabeni ezitshalwe ezinegciwane elilodwa kwadlulela emaqabunga, kwakha i-foci eyinhloko yesifo, engahlangana nokukhula okukhulu kwalesi sifo.
Ngakho-ke, kwezinye izifunda kungase kube nokushintshaniswa kwemijikelezo yezocansi kanye ne-asexual, kanti kwezinye - umjikelezo we-asexual kuphela.
Umsuka we-P. INFESTANS
I-P. infestans yavela eYurophu ekupheleni kwengxenye yokuqala yekhulu le-1991. Njengoba amazambane adabuka enyakatho-mpumalanga yeNingizimu Melika, kwakucatshangwa ukuthi i-parasite yalethwa esuka lapho yayiswa eYurophu ngesikhathi se-boom ye-saltpeter yaseChile. Kodwa-ke, ucwaningo olwenziwa esiteshini samazambane saseRockefeller Center eSigodini saseToluca, eMexico kwaphoqa ukuthi lo mbono ubhekwe kabusha (Niederhauser, 1993, XNUMX).
1. Esigodini sase-Toluca, izinhlobo zamazambane ezinama-tuberous zendawo (i-Solanum demissum, S. bulbocastanum, njll.) zinamasethi ahlukene ezakhi zofuzo zokumelana nokuma kuhlanganiswe nezinga eliphezulu lokumelana okungaqondile, okubonisa ukuvela kwemvelo okude ne-parasite. Izinhlobo zaseNingizimu Melika, okuhlanganisa namazambane ezitshalo, azinazo izakhi zofuzo.
2. Esigodini sase-Toluca, kutholakala izindawo ezingazodwa ezinezinhlobo zokukhwelana u-A1 no-A2, ngenxa yalokho inani labantu abaxubekile be-P. infestans lisakazeke kabanzi; kuyilapho ezweni lendabuko lamazambane alinyiwe, eNingizimu Melika, i-parasite isakazeka ngokuhlukana.
3. Esigodini saseToluca, kunezifo eziwumshayabhuqe ezinzima zaminyaka yonke zokulimaza kwakamuva. Ngakho-ke, phakathi kwabacwaningi baseNyakatho Melika (iNyuvesi yaseCornell), umbono mayelana neMesoamerica (IMelika Ephakathi) njengendawo yokuzalwa ye-phytophthora yamazambane uyasungulwa (Goodwin et al., 1994).
Abacwaningi baseNingizimu Melika abahambisani nalo mbono. Bakholelwa ukuthi amazambane atshaliwe kanye ne-parasite yayo i-P. infestans inezwe elivamile - i-Andes yaseNingizimu Melika. Basekela umbono wabo ngezifundo zamangqamuzana ekuhlaziyweni kwe-DNA polymorphisms ye-mitochondrial genome (mtDNA) kanye nezakhi zofuzo zenuzi i-RAS kanye ne-β-tubulin (Gomez-Alpizar et al., 2007). Babonisa ukuthi izinhlobonhlobo eziqoqwe ezingxenyeni ezihlukahlukene zomhlaba ziphuma emigqeni yokhokho emithathu ehlukene (bobathathu) etholakala e-Andes yaseNingizimu Melika. Ama-haplotype ase-Andean ayinzalo yemigqa emibili: izigaba zozalo oludala kakhulu lwe-mtDNA zitholakala ku-Solanaceae yasendle kusukela esigabeni se-Anarrhicomenum e-Ecuador, kuyilapho ukuhlukaniswa komugqa wesibili kuvamile kumazambane, utamatisi kanye ne-nightshade yasendle. E-Toluca, ngisho nama-haplotypes angandile aphuma ohlwini lozalo olulodwa kuphela, ngokuhlukahluka kofuzo kwezinhlobo ezivela ku-Toluca (imvamisa ephansi ye-allelic yamanye amasayithi aguquguqukayo) kuphakamisa umthelela oqinile womsunguli ngenxa yokukhukhuleka kwakamuva.
Ngaphezu kwalokho, kwatholakala uhlobo olusha lwe-P. andina e-Andes, ngokwe-morphologically kanye nezakhi zofuzo ezifana ne-P. infestans, ngokusho kwababhali, ikhomba i-Andes njengendawo eshisayo yokuhlola uhlobo lwePhytophthora. Ekugcineni, e-Europe nase-United States, ama-P. infestans ahlanganisa izinhlu zozalo zama-Andean, kuyilapho e-Toluca eyodwa kuphela.
Lokhu kushicilelwa kubangele ukuhlehla kweqembu labacwaningi abavela emazweni ahlukene, abenze umsebenzi omningi wokuhlola ukuze babuyekeze ucwaningo lwangaphambilini (Goss et al., 2014). Kulo msebenzi, okokuqala, ukulandelana kwe-DNA ye-microsatellite efundisayo kusetshenziswe ukutadisha ama-polymorphisms e-DNA; okwesibili, ukuze kuhlaziywe amaqoqo, izindlela zokufuduka, ukwehlukana kwesikhathi kwenani labantu, njll. kwasetshenziswa amamodeli athuthuke kakhulu (izibalo ze-F, izilinganiso ze-Bayesian, njll.) futhi, okwesithathu, ukuqhathanisa akusetshenziswanga kuphela uhlobo lwe-Andean P. andina, lapho kwasungulwa khona imvelo eyingxube (P. infestans x Phytophthora sp.) , kodwa futhi nezinhlobo ze-endmic zase-Mexican P. mirabilis, P. Ipomoeae, kanye ne-Phytophthora phaseoli - i-genetically close P. infestans efakwe ku-clade efanayo (Kroon et al., 2012). Ngenxa yalokhu kuhlaziya, kwaboniswa ngokusobala ukuthi ingxenye yempande yesihlahla se-phylogenetic yazo zonke izinhlobo zezinhlobo ze-Phytophthora ezithathwe ocwaningweni, ngaphandle kwe-hybrid P. Andi, ingeyezinhlobo zaseMexico, futhi ukugeleza kokufuduka kune isiqondiso iMexico - Andes, hhayi okuphambene nalokho, futhi ukuqala kwayo kuhambisana nekoloni yaseYurophu yeNew World (eminyakeni engu-300-600 edlule). Ngakho-ke, ukuvela kwezinhlobo ze-P. infestans, okukhethekile ngokunqotshwa kwamazambane, kwenzeka esikhungweni sesibili sofuzo sokwakheka kwama-nightshades angama-tuberous, i.e. eMelika Ephakathi.
I-Genome ye-P. INFESTANS
Ngo-2009, ithimba lamazwe ngamazwe lososayensi balandelanisa i-P infestans genome ephelele (Haas et al, 2009), usayizi wayo wawungu-240 MB. Lokhu kuphindwe izikhathi eziningana kunezinhlobo ezihlobene eduze i-P. sojae (95 Mb), okubangela ukubola kwezimpande zikabhontshisi wesoya, no-P. Ramorum (65 Mb), okuthinta izinhlobo zezihlahla ezibalulekile njenge-oki, i-beech nezinye. Idatha etholiwe ibonise ukuthi i-genome iqukethe inani elikhulu lamakhophi okulandelana okuphindaphindiwe - 74%. I-genome iqukethe izakhi zofuzo ze-protein-coding ezingu-17797, inqwaba yazo okuyizakhi zofuzo ezibandakanyeka ezinqubweni zamaselula, okuhlanganisa ukuphindaphinda kwe-DNA, ukuloba nokuhunyushwa kwamaprotheni.
Ukuqhathaniswa kwezakhi zofuzo zohlobo lwePhytophthora kwembule inhlangano engavamile ye-genome, ehlanganisa amabhlogo wokulandelana kwezakhi zofuzo ezilondoloziwe, lapho ukuminyana kwezakhi zofuzo kuphakeme kakhulu, futhi okuqukethwe kokulandelana okuphindaphindiwe kuphansi ngokuqhathaniswa, nezifunda ngazinye ezingalondoloziwe. ukulandelana kofuzo, okunofuzo oluncane kanye nokuqukethwe okuphezulu kwezifunda eziphindayo. Amabhulokhi e-Conservative ahlanganisa ama-70% (12440) azo zonke izinhlobo ze-P. infestans ze-protein-coding. Ngaphakathi kwamabhulokhi alondoloziwe, izakhi zofuzo zivame ukuhlukaniswa eduze nebanga eliphakathi nendawo elingu-604 bp. Ezindaweni eziphakathi kwamabhulokhi alondolozayo, ibanga le-intergenic likhulu (3700 bp) ngenxa yokwanda kokuminyana kwezinto eziphindayo. Izakhi zofuzo eziyimfihlo eziguqukayo zitholakala ezindaweni ezimpofu kakhulu.
Ukuhlaziywa kokulandelana kofuzo kwe-P. Infestans kwabonisa ukuthi cishe ingxenye eyodwa kwezintathu ye-genome ingeyezakhi ezishintshekayo. I-genome ye-P. infestans iqukethe imindeni ehluke kakhulu yama-transposons kunamanye ama-genome aziwayo. Iningi lama-P. Infestans transposons angamalungu omndeni wamaGypsy.
Inani elikhulu lemindeni ethile yofuzo ehilelekile ku-pathogenesis ikhonjwe ku-P. infestans genome. Ingxenye ebalulekile yazo ihlanganisa amaprotheni asebenzayo ashintsha i-physiology yesitshalo esiphethe futhi abe nesandla ekuthelelekeni kwaso. Ziwela ezigabeni ezimbili ezibanzi: ama-apoplastic effect, asebenza ezindaweni ezihlanganayo (ama-apoplast), nama-cytoplasmic effectors, angena kumaseli nge-haustoria. Ama-apoplastic effecters ahlanganisa ama-enzyme e-hydrolytic afihliwe njengama-proteases, i-lipase nama-glycosylases abhubhisa amaseli ezitshalo; ama-inhibitors wama-enzyme okuvikela izitshalo ezibambayo; kanye ne-necrotizing toxins njenge-Nep1-like proteins (NPLs) kanye nama-Pcf-like small cysteine-rich proteins (SCRs).
Izakhi zofuzo ze-P. infestans effector ziningi futhi ngokuvamile zikhulu kunezingezona ze-pathogenic. Aziwa kakhulu ama-cytoplasmic effectors i-RXLR ne-Crinkler (CNR). Izimpawu ezijwayelekile ze-cytoplasmic ze-oomycetes zingamaprotheni e-RXLR. Zonke izinhlobo zofuzo ze-RXLR ezitholwe kuze kube manje ziqukethe iqembu le-amino-terminal i-Arg-XLeu-Arg, lapho u-X eyi-amino acid. Njengomphumela wocwaningo, kwaphakanyiswa ukuthi kunezakhi zofuzo ezingu-563 ze-RXLR ku-P. infestans genome, okungama-60% ngaphezu kwe-P. sojae ne-P. ramorum. Cishe uhhafu wezakhi zofuzo ze-RXLR ku-P. infestans genome ziqondene nezinhlobo ezithile. Ama-effectors e-RXLR anezinhlobonhlobo eziningi zokulandelana. Phakathi kwabo, kuhlonzwe umndeni owodwa omkhulu nemincane engu-150. Ngokungafani ne-proteome eyinhloko, izakhi zofuzo ze-RXLR ngokuvamile zitholakala ezindaweni ezimpofu neziphindaphindayo ze-genome. Ama-elementi eselula anquma amandla alezi zifunda akhuthaza ukuhlanganisa kabusha kulawa majini.
Ama-Cytoplasmic CRN effectors ahlonzwe ekuqaleni emibhalweni ye-P. infestans encoding peptides eyenza i-necrosis yezicubu zezitshalo. Selokhu batholakala, kuncane okwaziwayo ngomndeni walaba benzi. Ukuhlaziywa kofuzo lwe-P. Infestans kwembule umndeni omkhulu wezakhi zofuzo ze-CRN ezingu-196, omkhulu kakhulu kunalowo we-P. sojae (100 CRN) kanye ne-P. ramorum (19 CRN). Njengama-RXLR, ama-CRN angamaprotheni ajwayelekile futhi aqukethe isizinda se-N-terminal LFLAK esilondolozwe kakhulu (ama-amino acid angama-50) kanye nesizinda esiseduze se-DWL esiqukethe izakhi zofuzo ezahlukene. Ama-CRN amaningi (60%) ane-peptide yesiginali.
Amathuba ama-CRN ahlukahlukene okuphazamisa izinqubo zamaselula esitshalweni sokusingatha kuye kwafundwa. Lapho kuhlaziywa i-necrosis yezitshalo, ukususwa kwamaprotheni e-CRN2 kwenza kwaba nokwenzeka ukuhlonza isifunda se-C-terminal esihlanganisa ama-amino acid angu-234 (izikhundla 173-407, isizinda se-DXG) futhi kubangela ukufa kweseli. Ukuhlaziywa kwezakhi zofuzo ze-CRN ze-P. infestans kwembule izifunda ezine ezihlukene ze-C-terminal, ezibangela nokufa kwamaseli ngaphakathi kwesitshalo. Lokhu kufaka phakathi izizinda ze-DC ezisanda kukhonjwa (i-P. Infestans inezakhi zofuzo ezingu-18 kanye nama-pseudogenes angu-49), kanye nezizinda ze-D2 (14 kanye ne-43) ne-DBF (2 kanye ne-1) ezifana ne-protein kinases. Amaprotheni ezizinda ze-CRN ezivezwe esitshalweni ayalondolozwa (ngokungabikho kwama-peptide esignali) esitokisini sesitshalo futhi akhuthaze ukufa kweseli ngomshini we-intracellular. Okunye ukulandelana okungu-255 okuqukethe izizinda ze-CRN okungenzeka ukuthi akusebenzi njengezakhi zofuzo.
Ukwenyuka kwenombolo nosayizi wemindeni yezakhi zofuzo ze-RXLR kanye ne-CRN kungenzeka kube ngenxa yokuhlanganisa okungeyona i-allelic homologous kanye nokuphindaphindeka kofuzo. Naphezu kweqiniso lokuthi i-genome iqukethe inani elikhulu lezakhi zeselula ezisebenzayo, akukabikho ubufakazi obuqondile bokudluliselwa kwezakhi zofuzo ze-effector.
Izindlela ezisetshenziswa ocwaningweni lwesakhiwo sabantu
Ucwaningo lwesakhiwo sofuzo sabantu okwamanje lusekelwe ekuhlaziyweni kwamasiko ahlanzekile ezinhlobo zawo. Ukuhlaziywa kwenani labantu ngaphandle kokuhlukanisa izitshalo ezihlanzekile nakho kwenziwa ngezinhloso ezithile, ezifana, ngokwesibonelo, ukufunda ulaka lwesibalo sabantu noma ukuba khona kwezinhlobo ezimelana nezibulala-sikhunta kuso (Filippov et al., 2004; Derevyagina et al., 1999). Lolu hlobo locwaningo lubandakanya ukusetshenziswa kwezindlela ezikhethekile, incazelo yazo engaphezu kobubanzi balokhu kubuyekezwa. Ukuze kuhlaziywe ngokuqhathaniswa kwezinhlobo, kusetshenziswe izindlela eziningi, ngokusekelwe kokubili ekuhlaziyweni kwesakhiwo se-DNA kanye nocwaningo lokubonakaliswa kwe-phenotypic. Ukuhlaziywa okuqhathanisayo kwenani labantu kufanele kubhekane nenani elikhulu lezihlukanisi, okubeka izidingo ezithile ezindleleni ezisetshenziswayo. Ngokufanelekile, kufanele bahlangabezane nezidingo ezilandelayo (Cooke, Lees, 2004, Mueller, Wolfenbarger, 1999):
- kube ishibhile, kube lula ukukusebenzisa, akudingi izindleko zesikhathi ezibalulekile, kusekelwe kubuchwepheshe obutholakala ngokuvamile (isibonelo, i-PCR);
- kumele akhiqize inani elikhulu ngokwanele lezimpawu ezizimele ze-codominant marker;
- babe nokuphindaphinda okuphezulu;
- sebenzisa inani elincane lezicubu okufanele zihlolwe;
- kucaciswe ku-substrate (ukungcola okukhona kusiko akufanele kuthinte imiphumela);
- ayidingi ukusetshenziswa kwezinqubo eziyingozi namakhemikhali anobuthi obukhulu.
Ngeshwa, azikho izindlela ezihambisana nawo wonke amapharamitha angenhla. Ukucwaninga okuqhathanisayo kwezinhlobo zesikhathi sethu, izindlela zisetshenziswa ngokususelwa ekuhlaziyweni kwezici ze-phenotypic: i-virulence ezinhlobonhlobo zamazambane notamatisi (izinhlanga zamazambane notamatisi), uhlobo lokukhwelana, i-spectra ye-peptidase kanye ne-glucose-6-phosphate isomerase isoenzymes, kanye ekuhlaziyweni kwesakhiwo se-DNA: ubude be-polymorphism restriction fragment (RFLP), evame ukulekelelwa nge-hybridization probe RG 57, ukuhlaziya okuphindaphindwayo kwe-microsatellite (SSR ne-InterSSR), ukukhulisa ngama-random primers (RAPD), ukukhulisa izingcezwana zokuthibela (AFLP) , ukukhulisa ngeziqalo ezihambisana nokulandelana kwezakhi zeselula (isibonelo, i-Inter SINE PCR), ukunqunywa kwama-haplotypes e-DNA emitochondrial.
Izincazelo ezimfushane zezindlela zocwaningo lokuqhathanisa lwezinhlobo ezisetshenziswa emsebenzini nama-P. infestans
Izimpawu zomaka we-Phenotypic
"Potato" izinhlanga
Izinhlanga "zamazambane" ziwuphawu oluvame ukucwaningwa futhi lusetshenziswe. Izinhlanga "zamazambane alula" zinesakhi esisodwa se-potato virulence, "eziyinkimbinkimbi" - okungenani ezimbili. UBlack et al. (1953), efingqa yonke imininingwane yabo, bathola ukuthi umjaho we-phytophthora uyakwazi ukuthelela izitshalo ngofuzo/izakhi zofuzo ezihambisana ne-P. infestans virulence gene / gene, futhi bathola izinhlanga 1, 2, 3, kanye 4 ezithelela izitshalo ngezakhi zofuzo u-R1, u-R2, u-R3 no-R4, ngokulandelana, i.e. ukusebenzisana phakathi kwe-parasite kanye nomsingathi kwenzeka ngesisekelo sofuzo nge-gene. Ngaphezu kwalokho, uBlack, ngokubamba iqhaza kukaGallegly noMalcolmson, wathola izinhlobo zofuzo u-R5, R6, R7, R8, R9, R10 no-R11, kanye nezinhlanga ezihambisanayo (Black, 1954; Black & Gallegly, 1957; Malcolmson & Black) , 1966; Malcolmson, 1970).
Kukhona indikimba yedatha emayelana nokwakheka kohlanga lwe-pathogen evela ezifundeni ezahlukahlukene. Ngaphandle kokuhlaziya le datha ngokuningiliziwe, sizokhombisa inkambiso ejwayelekile kuphela: lapho kwasetshenziswa izinhlobo ezinezakhi zofuzo ezintsha zokumelana noma inhlanganisela yazo, ekuqaleni kwaba khona ukuwohloka okuthile kokulimaza sekwephuzile, kodwa kwase kuvela izinhlanga ezinezakhi zofuzo ezihambisanayo futhi zakhethwa kanye nokuqubuka. isifo esibi sekwephuzile saqala kabusha. Ukuhlukumezeka okuqondile kwezakhi zofuzo ezi-4 zokuqala zokumelana (R1-R4) kwakungavamile ukubonwa emaqoqweni aqoqwe ngaphambi kokwethulwa ekutshalweni kwezinhlobo ezinalezi zakhi zofuzo, kodwa inani lezinhlobo ezinohlonze landa kakhulu lapho i-pathogen ihlukunyezwa ezinhlobonhlobo eziphethe lezi zakhi zofuzo. UGenes 5-11, ngakolunye uhlangothi, zazivame kakhulu emaqoqweni (Shaw, 1991).
Ucwaningo lwesilinganiso sezinhlanga ezahlukene ngesikhathi sokukhula, olwenziwa ngasekupheleni kwawo-1980, lwabonisa ukuthi ekuqaleni kokukhula kwalesi sifo, ama-clones anolaka oluphansi kanye nezakhi zofuzo ezi-1-2 ze-virulence zigcwele abantu.
Ngaphezu kwalokho, njengoba ukubola kwakamuva kukhula, ukugxila kwama-clone okuqala kuncipha futhi nenani lezinhlanga “eziyinkimbinkimbi” ezinolaka oluphezulu liyakhula. Ukwenzeka kwalokhu kugcina kufinyelela ku-100% ekupheleni kwesizini. Lapho ugcina izilimo eziyizigaxa, kuba nokuncipha kolaka kanye nokulahlekelwa izakhi zofuzo ze-virulence. I-dynamics yokushintshwa kwe-clone ingenzeka ngezinhlobo ezahlukene ngezindlela ezahlukene (Rybakova & Dyakov, 1990). Kodwa-ke, izifundo zethu ngo-2000-2010 zabonisa ukuthi izinhlanga eziyinkimbinkimbi zitholakala kusukela ekuqaleni kwe-epiphytotics phakathi kwezinhlobo ezihlukaniswe kokubili amazambane notamatisi. Lokhu mhlawumbe kungenxa yezinguquko kubantu base-P. Infestans eRussia.
Ngo-1988-1995, ukuvela "kwezindawo eziphakeme kakhulu" nazo zonke noma cishe zonke izakhi zofuzo ze-virulence ezifundeni ezahlukene zafinyelela ku-70-100%. Lesi simo saphawulwa, isibonelo, eBelarus, ezindaweni zaseLeningrad naseMoscow, eNyakatho Ossetia naseJalimane (Ivanyuk et al., 2002a, 2002b; Politiko, 1994; Schober-Butin et al., 1995).
"Utamatisi" izinhlanga
Ezitshalweni zikatamatisi, zimbili kuphela izinhlobo zofuzo zokumelana nokubola okuphuzile ezitholakele - i-Ph2 (Gallegly & Marvell, 1) kanye ne-Ph1955 (Al-Kherb, 2). Njengasendabeni yezinhlanga zamazambane, ukusebenzisana phakathi kukatamatisi kanye ne-P. infestans kwenzeka ngesisekelo sofuzo. Umjaho we-T1988 uthelela izinhlobo ezingenazo izakhi zofuzo (iningi lezinhlobo ezisetshenziswayo ezithengiswayo), umjaho we-T0 uthelela izinhlobo zofuzo lwe-Ph1 (Ottawa), kanti umjaho we-T1 uthelela izinhlobo ngohlobo lwe-Ph2.
E-Russia, cishe i-T0 kuphela yatholakala kumazambane; kumatamatisi ekuqaleni kwesizini, i-T0 yanqoba, kodwa kamuva yathathelwa indawo umjaho we-T1 (Dyakov et al., 1975, 1994). Ngemuva kuka-2000, i-T1 kumazambane emiphakathini eminingi yaqala ukwenzeka ekuqaleni kwenkathi ye-epiphytotic. E-United States, izinhlobo zamazambane zazingenalo i-pathogenic kutamatisi, kanye nezinhlanga i-T0, i-T1, ne-T2, kuyilapho i-T1 ne-T2 yayidla utamatisi (Vartanian & Endo, 1985; Goodwin et al., 1995).
Uhlobo lokukhwelana
Ukuze wenze ucwaningo, izinhlobo zomhloli (inkomba) ezinezinhlobo zokukhwelana ezaziwayo - i-A1 ne-A2 iyadingeka. I-test isolate ifakwe nabo ngamabili ezitsheni ze-Petri nge-oat agar medium. Ngemuva kokufukamela izinsuku eziyi-10, amapuleti ahlolwe ukuthi akhona yini noma angekho ama-oospores endaweni yokuxhumana yezinkinga. Kunezinketho ezi-4: uhlobo lohlobo lokukhwelana lwe-A1, uma lwakha ama-oospores ngesihloli se-A2, kuya ku-A2, uma lwakha ama-oospores ngesihloli se-A1, luye ku-A1A2, uma lwakha ama-oospores nabo bobabili abahloli, noma luyinyumba 00), uma ingenzi ama-oospores angenawo umhloli (amaqembu amabili okugcina ayivelakancane).
Ukuze kutholakale ngokushesha izinhlobo zokukhwelana, kwenziwa imizamo yokuhlonza izifunda ze-genome ezihlotshaniswa nohlobo lokukhwelana, ngenhloso yokuqhubeka nokusetshenziswa kwazo ukuze kutholwe uhlobo lokukhwelana kwe-PCR. Esinye sezivivinyo zokuqala eziyimpumelelo zokuhlonza isiza esinjalo senziwe abacwaningi baseMelika (Judelson et al., 1995). Besebenzisa indlela ye-RAPD, bakwazile ukuhlonza isifunda se-W16 esihlotshaniswa nohlobo lokukhwelana enzalweni yama-isolate amabili awela, futhi baklama ipheya ye-24-bp primers yokukhulisa kwayo (W16-1 (5'-AACACGCACAAGGCATATAAATGTA-3 ') kanye ne-W16-2 (5' -GCGTAATGTAGCGTAACAGCTCTC-3 ') Ngemva kokukhawulelwa komkhiqizo we-PCR nge-enzyme evimbela i-HaeIII, kube nokwenzeka ukuhlukanisa okukodwa ngezinhlobo zokumatanisa u-A1 no-A2.
Omunye umzamo wokuthola izimpawu ze-PCR ukuze kutholwe izinhlobo zokukhwelana wenziwa abacwaningi baseKorea (Kim, Lee, 2002). Bahlonze imikhiqizo ethile besebenzisa indlela ye-AFLP. Njengomphumela, ipheya yeziqalo PHYB-1 (phambili) (5'-GATCGGATTAGTCAGACGAG-3 ') kanye ne-PHYB-2 (5'-GCGTCTGCAAGGCGCATTTT-3') zakhiwe, okuvumela ukukhuliswa okukhethiwe kwesifunda sofuzo esihlotshaniswa nokuhlangana kwe-A2. uhlobo. Ngokulandelayo, baqhubeka nalo msebenzi futhi baklama ama-primers 5 'AAGCTATACTGGGACAGGGT-3' (INF-1, phambili) kanye ne-5'-GCGTTCTTTCGTATTACCAC-3 '(INF-2), okuvumela ukukhuliswa okukhethiwe kwesimo sesifunda se-Mat-A1 sezinhlobo zokukhwelana. uhlobo A1. Ukusetshenziswa kokuxilongwa kwe-PCR kwezinhlobo zokukhwelana kubonise imiphumela emihle lapho kufundwa imiphakathi yama-P. infestans e-Czech Republic (Mazakova et al., 2006), Tunisia (Jmour, Hamada, 2006) nezinye izifunda. Elabhorethri yethu (Mytsa, Elansky, unpublished), 34 P. infestans strains ehlukaniswe namazambane agulayo kanye nezitho zotamatisi ezindaweni ezahlukene zaseRussia (Kostroma, Ryazan, Astrakhan, Moscow oblasts) zahlaziywa. Imiphumela yokuhlaziywa kwe-PCR kusetshenziswa iziqalo ezithile ezingaphezu kuka-90% ziqondane nemiphumela yokuhlaziywa kohlobo lokukhwelana ngendlela yendabuko endaweni yezakhi.
Ithebula 1. Ukuhlukahluka kokumelana ngaphakathi kwe-Sib 1 clone (Elansky et al., 2001)
Indawo yokuqoqa isampula | Inani lama-isolate ahlaziywe | Inombolo yokuzwela (S), ukumelana ngokubuthakathaka (SR) kanye nokumelana (R), ama-pcs (%) | ||
S | SR | R | ||
G. Vladivostok | 10 | 1 (10) | 4 (40) | 5 (50) |
G. Chita | 5 | 0 | 0 | 5 (100) |
G. Irkutsk | 9 | 9 (100) | 0 | 0 |
G. Krasnoyarsk | 13 | 12 (92) | 1 (8) | 0 |
Idolobha laseYekaterinburg | 15 | 8 (53) | 1 (7) | 6 (40) |
O. Sakhalin | 66 | 0 | 0 | 66 (100) |
Isifunda sase-Omsk | 18 | 0 | 0 | 18 (100) |
Ukumelana ne-Metalaxyl njengomaka wabantu
Ekuqaleni kwawo-1980, ukuqubuka okunamandla kokulimaza sekwephuzile okubangelwa izinhlobo ze-infestans ezimelana ne-metaxyl-resistant P. kwaphawulwa ezifundeni ezihlukahlukene. Amapulazi amazambane emazweni amaningi alahlekelwe kakhulu (Dowley & O'Sullivan, 1981; Davidse et al., 1983; Derevyagina, 1991). Kusukela ngaleso sikhathi, emazweni amaningi omhlaba, ukuqapha okuqhubekayo kokuvela kwezinhlobo ezimelana ne-phenylamide ku-P. infestans labantu kuye kwenziwa. Ngaphezu kokuhlolwa okungokoqobo kwamathemba okusetshenziswa kwezidakamizwa eziqukethe i-phenylamide, ukwakha uhlelo lwezinyathelo zokuzivikela nokubikezela ama-epiphytoties, ukumelana nale mithi kuye kwaba esinye sezici zomaka ezisetshenziswa kabanzi ekuhlaziyeni okuqhathanisayo kwezibalo zaleli pathogen. Kodwa-ke, ukusetshenziswa kokumelana ne-metaxyl ezifundweni zokuqhathanisa zabantu kufanele kwenziwe kucatshangelwa iqiniso lokuthi: 1 - isisekelo sofuzo sokumelana asikanqunywa ngokunembile, 2 - ukumelana ne-metaxyl kuyisici esincike ngokukhetha esingashintsha. kuye ngokuthi ukusetshenziswa phenylamides, 3 - ezahlukene degree of ukuzwela metalaxyl amagciwane ngaphakathi komugqa owodwa clonal (ithebula. 1).
I-spectra ye-isozyme
Omaka be-Isozyme ngokuvamile abazimele ezimweni zangaphandle, babonisa ifa le-Mendelian futhi bahlangene, okuvumela ukuhlukanisa phakathi kwama-homo- nama-heterozygote. Ukusetshenziswa kwamaphrotheni njengamakaki wofuzo kwenza kube nokwenzeka ukuhlonza kokubili ukuhlelwa kabusha okukhulu kwezakhi zofuzo, okuhlanganisa ukuguqulwa kwe-chromosomal ne-genomic, kanye nokushintshwa okukodwa kwe-amino acid.
Ucwaningo lwe-Electrophoretic lwamaprotheni lubonise ukuthi ama-enzyme amaningi akhona ezinto eziphilayo ngendlela yama-fractions amaningana ahluke ekuhambeni kwe-electrophoretic. Lezi zingxenyana ziwumphumela wokubhala ngekhodi izinhlobo eziningi zama-enzyme nge-loci ehlukene (ama-isozyme noma ama-isozyme) noma ngama-allele ahlukene we-locus efanayo (ama-allozyme noma ama-alloenzymes). Okusho ukuthi, ama-isozyme ayizinhlobo ezahlukene ze-enzyme eyodwa. Izinhlobo ezihlukene zinomsebenzi ofanayo we-catalytic, kodwa zihluka kancane ekushintsheni okukodwa kwe-amino acid ku-peptide nephetheyo. Umehluko onjalo wembulwa ngesikhathi se-electrophoresis.
Lapho kufundwa izinhlobo ze-P. infestans, i-spectra ye-isoenzymes yamaprotheni amabili, i-peptidase ne-glucose-6-phosphate isomerase, isetshenziswa (le enzyme i-monomorphic kubantu baseRussia, ngakho-ke, izindlela zocwaningo lwayo azivezwa kulo msebenzi). Ukuze uwahlukanise abe ama-isozyme emkhakheni kagesi, amalungiselelo amaprotheni ahlukaniswe nezinto eziphilayo ezihloliwe asetshenziswa epuleti yejeli ebekwe endaweni kagesi. Izinga lokusatshalaliswa kwamaprotheni ngamanye kujeli lincike ekukhokhiseni kanye nesisindo samangqamuzana, ngakho-ke, endaweni kagesi, ingxube yamaprotheni ihlukaniswa ibe yizingxenyana ngazinye, ezingabonwa kusetshenziswa odayi abakhethekile.
Ucwaningo lwe-peptidase isoenzymes lwenziwa ku-cellulose acetate, isitashi noma ama-polyacrylamide gels. Okulula kakhulu indlela esekelwe ekusetshenzisweni kwamajeli e-cellulose acetate akhiqizwa yi-Helena Laboratories Inc. Ayidingi inani elikhulu lezinto zokuhlola, ivumela umuntu ukuthi athole amabhande aphikisanayo ku-gel ngemuva kwe-electrophoresis kokubili i-enzyme loci, ukuqaliswa kwayo akudingi izindleko ezinkulu zesikhathi nempahla (Fig. 2).
Isiqephu esincane se-mycelium sidluliselwa ku-microtube engu-1,5 ml, amaconsi angu-1-2 wamanzi a-distilled engeziwe kuwo. Ngemuva kwalokho, isampula i-homogenized (isibonelo, nge-drill kagesi ene-attachment epulasitiki efanelekile ku-microtube) futhi ifakwe imizuzwana engu-25 ku-centrifuge ku-13000 rpm. Kusuka ku-microtube ngayinye, 8 μl. i-supernatant idluliselwa epuleti lokufaka isicelo.
Ijeli ye-cellulose acetate ikhishwa esitsheni se-buffer, isulwe phakathi kwamakhasi amabili ephepha lokuhlunga bese ibekwa nongqimba olusebenzayo phezu kwesisekelo sepulasitiki sofake isicelo. Isixazululo esivela epuleti sidluliselwa ngumfakisicelo ku-gel izikhathi ezingu-2-4. Ijeli idluliselwa ekamelweni le-electrophoresis,
Ithebula 2. Ukwakhiwa kwesisombululo esisetshenziselwa ukungcolisa ijeli ye-cellulose i-acetate ekuhlaziyeni ama-isoenzymes e-peptidase, ithonsi lopende (i-bromophenol eluhlaza okwesibhakabhaka) libekwe emaphethelweni ejeli.
I-TRIS HCl, 0,05M, Ph 8,0 2 ml
Peroxidase, 1000 U / ml 5 amaconsi
o-dianisidine, 4 mg / ml 8 amaconsi
MgCl2, 20 mg / ml 2 amaconsi
Gly-Leu, 15 mg / ml 10 amaconsi
L-amino-acid oxidase, 20 u / ml 2 amaconsi
I-Electrophoresis yenziwa imizuzu engama-20. ku-200 V. Ngemuva kwe-electrophoresis, i-gel idluliselwa etafuleni lokudweba futhi igcotshwe ngesisombululo esikhethekile sokudweba (Ithebula 2). I-10 ml ye-1,6% ye-DIFCO agar incibilika kuqala kuhhavini ye-microwave, ipholiswe ku-60 ° C, emva kwalokho i-2 ml ye-agar ihlanganiswe nengxube kapende bese ithululelwa ejeli. Imivimbo ibonakala phakathi kwemizuzu engu-15-20. I-reagent ye-L-amino-acid oxidase yengezwa ngaphambi nje kokuxuba isixazululo ne-agar encibilikisiwe.
Kubantu baseRussia, i-Pep 1 locus imelwe yi-genotypes 100/100 kanye ne-92/100. I-Homozygote 92/92 iyivelakancane kakhulu (cishe i-0,1%). I-Locus Rehr 2 imelelwa ama-genotypes amathathu 100/100, 100/112, kanye no-112/112, futhi zonke izinhlobo ezi-3 zivame kakhulu (Elanky kanye ne-Smirnov, 2003, Fig. 2).
Ucwaningo lwe-genome
I-polymorphism yobude bengcezu ekhawulelwe ne-hybridization elandelayo (RFLP-RG 57)
Isamba se-DNA siphathwa nge-Eco R1 restriction enzyme, izingcezu ze-DNA zihlukaniswa yi-agarose gel electrophoresis. I-Nuclear DNA inkulu kakhulu futhi inokulandelana okuningi okuphindaphindayo; ngakho-ke, ukuhlaziya okuqondile kwezingcezu eziningi ezitholwe isenzo sokuvinjelwa kwama-enzyme kunzima ukukufeza. Ngakho-ke, izingcezwana ze-DNA ezihlukaniswa kujeli zidluliselwa kulwelwesi olukhethekile futhi lusetshenziselwe ukuhlanganisa nge-RG 57 probe, ehlanganisa ama-nucleotide anelebula amalebula e-radioactive noma e-fluorescent. Le probe ixubene ngokulandelana kwe-genomic ephindaphindayo (Goodwin et al., 1992, Forbes et al., 1998). Ngemuva kokuboniswa ngeso lengqondo kwemiphumela ye-hybridization ezintweni ezikhanyayo noma ezinemisebe, iphrofayili ye-multi-locus ye-hybridization (izigxivizo zeminwe), emelwe yizingcezu ezingama-25-29, iyatholakala (Forbes et al., 1998). Inzalo ye-Asexual (clonal) izoba namaphrofayili afanayo. Ngokuhlelwa kwamabhendi ku-electrophoretogram, ukufana nokuhluka kwezinto eziphilayo eziqhathaniswayo kuyahlulelwa.
I-Mitochondrial DNA haplotypes
Emangqamuzaneni amaningi e-eukaryotic, i-mtDNA yethulwa ngendlela ye-molecule ye-DNA eyindilinga enemicu ephindwe kabili, leyo, ngokungafani nama-chromosome enuclear yamangqamuzana e-eukaryotic, iphindaphinda i-semi-conservative futhi ayihlobene nama-molecule amaprotheni.
I-genome ye-mitochondrial ye-P. infestans yalandelana, futhi imisebenzi eminingana yanikelwa ekuhlaziyeni ubude bezingcezu ezivinjiwe (Carter et al, 1990, Goodwin, 1991, Gavino, Fry, 2002). Ngemva kokuthi u-Griffith noShaw (1998) benze indlela elula nesheshayo yokunquma ama-haplotypes e-mtDNA, lo maka kube ngomunye odume kakhulu ocwaningweni lwe-P. Infestans. ) kanye ne-EcoR2 (isiqephu sesibili). Indlela ivumela ukuhlonza ama-haplotype angu-2: Ia, IIa, Ib, IIb. Uhlobo II luhlukile ohlotsheni I ngokuba khona kokufaka okungu-4 bp ngosayizi kanye nendawo ehlukile yezindawo zokukhawulela ezifundeni ze-P4 ne-P3 (Fig. 1).
Kusukela ngo-1996, phakathi kwezinhlobo eziqoqwe endaweni yaseRussia, kwaphawulwa kuphela ama-haplotype u-Ia no-IIa (Elansky et al., 2001, 2015). Zingabonakala ngemuva kokuhlukaniswa kwemikhiqizo yokuvinjelwa nge-primer F2-R2 ensimini kagesi (Fig. 4, 5). Izinhlobo ze-mtDNA zisetshenziswa ekuhlaziyeni okuqhathanisayo kwezinhlobo nezibalo zabantu. Emisebenzini eminingana, izinhlobo ze-DNA ye-mitochondrial zazisetshenziselwa ukuhlukanisa imigqa ye-clonal kanye ne-passportize P. infestans isolate (Botez et al., 2007; Shein et al., 2009). Kusetshenziswa indlela ye-PCR-RFLP, kwafinyelelwa esiphethweni sokuthi i-mtDNA ihlukile kuhlobo olufanayo lwe-P. infestans (Elansky and Milyutina, 2007). Izimo zokukhulisa: 1x (500 sec. 94 ° C), 40x (30 sec. 90 ° C, 30 sec. 52 ° C, 90 sec. 72 ° C); 1x (5 amaminithi. 72 ° C). Ingxube yokusabela: (20 μl): 0,2 U Taq DNA polymerase, 1x 2,5 mM MgCl2-Taq buffer, 0,2 mM i-dNTP ngayinye, 30 pM primer kanye no-5 ng we-DNA ehlaziywe, amanzi ahlukanisiwe - afika ku-20 μl.
Ukuvinjelwa komkhiqizo we-PCR kwenziwa amahora angu-4-6 ekushiseni kuka-37 ° C. Ingxube yokukhawulela (20 μl): 10x MspI (2 μl), 10x isikhawulo sebhafa (2 μl), amanzi akhishiwe (6 μl), umkhiqizo we-PCR (10 μl).
Ithebula 3. Iziqalo ezisetshenziselwa ukukhulisa izifunda ze-mtDNA polymorphic
I-Locus | I-Primer | Ubude be-primer nokubekwa | Ubude bomkhiqizo we-PCR | Khawulela |
---|---|---|---|---|
P2 | F2: 5'- TTCCCTTTGTCCTCTACCGAT | 21; 13619-13639 | 1070 | MspI |
R2: 5'- TTACGGCGGTTTAGCACATACA | 22; 14688-14667 | |||
P4 | F4: 5'- TGGTCATCCAGAGGTTTATGTT | 22; 9329-9350 | 964 | I-EcoRI |
R4:5 - CCGATACCGATACCAGCACCAA | 22; 10292-10271 |
I-Random primer amplification (RAPD)
Lapho kwenziwa i-RAPD, i-primer eyodwa (ngezinye izikhathi iziqalo ezimbalwa ngesikhathi esisodwa) isetshenziswa ngokulandelana kwe-nucleotide okungahleliwe, ngokuvamile ama-nucleotide angu-10 ubude, nokuqukethwe okuphezulu (kusuka ku-50%) wama-nucleotide e-GC kanye nezinga lokushisa eliphansi (cishe 35). ° C). Iziqalo ezinjalo "zihlala" kumasayithi amaningi ahambisanayo ku-genome. Ngemuva kokukhulisa, kutholwa inani elikhulu lama-amplicons. Inombolo yabo incike ku-primer (s) esetshenzisiwe kanye nezimo zokusabela (i-MgCl2 ukugxilisa kanye nezinga lokushisa lokuphefumula).
Ukubona ngeso lengqondo ama-amplicons kwenziwa nge-distillation ku-polyacrylamide noma i-agarose gel. Lapho wenza ukuhlaziywa kwe-RAPD, kuyadingeka ukuqapha ngokucophelela ukuhlanzeka kwezinto ezihlaziywe, kusukela ukungcoliswa nezinye izinto eziphilayo kungabangela ukwanda okukhulu kwenani lezinto zobuciko, okukhona kuzo ezimbalwa ngisho nasekuhlaziyeni izinto ezihlanzekile (Perez et al, 1998). Ukusetshenziswa kwale ndlela ekutadisheni i-P. infestans genome kubonakala emisebenzini eminingi (Judelson, Roberts, 1999, Ghimire et al., 2002, Carlisle et al., 2001). Ukukhethwa kwezimo zokusabela kanye nama-primers (ama-primer angu-51 10-nucleotide afundwa) anikezwe esihlokweni sika-Abu-El Samen et al., (2003).
I-Microsatellite Repeat Analysis (SSR)
Ukuphinda kwe-Microsatellite (ukuphindaphinda kokulandelana okulula, i-SSR) kuphinda ngokulandelana okufushane ukulandelana kwe-1-3 (kwesinye isikhathi kufika ku-6) ama-nucleotide akhona ku-genomes yenuzi yawo wonke ama-eukaryote. Inani lokuphindaphinda okulandelanayo lingahluka kusuka ku-10 kuye ku-100. I-Microsatellite loci yenzeka ngefrikhwensi ephezulu kakhulu futhi isatshalaliswa ngokulinganayo kuyo yonke i-genome (Lagercrantz et al., 1993). I-Polymorphism yokulandelana kwe-microsatellite ihlotshaniswa nomehluko enanini lokuphindaphinda kwe-motif eyisisekelo. Izimpawu ze-Microsatellite ziyi-codominant, okwenza kube nokwenzeka ukuzisebenzisa ukuhlaziya ukwakheka kwenani labantu, ukunquma ukuhlobana, izindlela zokufuduka zama-genotypes, njll. Phakathi kwezinye izinzuzo zalezi zomaka, umuntu kufanele aqaphele i-polymorphism yazo ephakeme, i-polymorphism yazo enhle, i-reproducibility enhle, ukungathathi hlangothi, kanye nekhono lokwenza ukuhlaziya nokuhlola okuzenzakalelayo.Ukuhlaziywa kwe-polymorphism yokuphindaphinda kwe-microsatellite kwenziwa ukukhuliswa kwe-PCR kusetshenziswa ama-primer ahambisana nokulandelana okuyingqayizivele okuphambana ne-microsatellite loci. . Ekuqaleni, ukuhlaziywa kwenziwa ngokuhlukaniswa kwemikhiqizo yokusabela kujeli ye-polyacrylamide. Kamuva, abasebenzi benkampani ye-Applied Biosystems bahlongoza ukusebenzisa ama-primer anelebuli ekhanyayo ngokutholwa kwemikhiqizo yokusabela kusetshenziswa umtshina we-laser othomathikhi (Diehl et al., 1990), bese kuba yizilandeleli ze-DNA ezizenzakalelayo (Ziegle et al., 1992). Ukulebula ama-primer anamadayi ahlukahlukene e-fluorescent kwenza kube nokwenzeka ukuhlaziya izimpawu ezimbalwa ngesikhathi esisodwa emzileni owodwa futhi, ngokufanelekile, kukhulisa kakhulu umkhiqizo wendlela futhi kwandise ukunemba kokuhlaziywa.
Izincwadi zokuqala ezinikelwe ekusetshenzisweni kokuhlaziywa kwe-SSR ocwaningweni lwe-P. infestans zavela ekuqaleni kwawo-2000. (Knapova, Gisi, 2002). Akubona bonke omaka abahlongozwe ababhali ababonisa izinga elanele le-polymorphism, nokho, ababili babo (4B kanye ne-G11) bafakwe kusethi yezimpawu ze-SSR eziyi-12 ezihlongozwe ngu-Lees et al. (2006) futhi zamukelwa kamuva kunethiwekhi yocwaningo ye-Eucablight. (www.eucablight .org) njengezinga le-P. infestans. Eminyakeni embalwa kamuva, ucwaningo lwanyatheliswa ekudalweni kwesistimu yokuhlaziywa kwe-multiplex ye-P. infestans DNA esekelwe kumaka we-SSR ayisishiyagalombili (Li et al., 2010). Okokugcina, ngemva kokuhlola bonke omaka abahlongoziwe ngaphambili nokukhetha ulwazi oluningi kakhulu kubo, kanye nokuthuthukisa ama-primers, amalebula e-fluorescent, nezimo zokukhulisa, ithimba elifanayo lababhali lethule uhlelo lokuhlaziya lwe-multiplex lwesinyathelo esisodwa oluhlanganisa izimpawu ze-12 (Ithebula 4; Li et al., 2013a). Ama-primer asetshenziswe kulolu hlelo akhethiwe futhi alebula okukodwa komaka abane be-fluorescent (FAM, VIC, NED, PET) ukuze ububanzi bosayizi we-allele weziqalisi ezinamalebula afanayo bungadluleli.
Ababhali benze ukuhlaziya ku-PTC200 amplifier (MJ Research, USA) besebenzisa i-QIAGEN multiplex PCR kits noma i-QIAGEN Typeit Microsatellite PCR kits. Umthamo wenhlanganisela yokusabela wawuyi-12.5 μL. Izimo zokukhulisa zazimi kanje: ku-QIAGEN multiplex PCR: 95 ° C (15 min), 30x (95 ° C (20 s), 58 ° C (90 s), 72 ° C (60 s), 72 ° C ( 20 min); ye-QIAGEN Type-it Microsatellite PCR): 95 ° C (5 min), 28x (95 ° C (30 sec), 58 ° C (90 sec), 72 ° C (20 sec), 60 ° C (30 imiz).
Ukuhlukaniswa nokubonwa kwemikhiqizo ye-PCR kwenziwa kusetshenziswa i-ABI3730 automatic capillary DNA analyzer (Applied Biosystems).
Ithebula 4. Izimpawu zomaka abangu-12 abajwayelekile be-SSR abasetshenziselwa i-genotyping P. Infestans (Li et al., 2013a)
Isihloko | Inombolo yama-alleles | Ibanga losayizi i-alleles (bp) | Ama-Primers |
I-PiG11 | 13 | 130-180 | F: NED-TGCTATTTATCAAGCGTGGG R: GTTTCAATCTGCAGCCGTAAGA |
UPi02 | 4 | 255-275 | F: NED-ACTTGCAGAACTACCGCCC R: GTTTGACCACTTTCCTCGGTTC |
I-PinfSSR11 | 4 | 325-360 | F: NED-TTAAGCCACGACATGAGCTG R: GTTTAGACAATTGTTTTGTGGTCGC |
D13 | 16 | 100-185 | F: FAM-TGCCCCCTGCTCACTC R: GCTCGAATTCATTTTTCAGACTTG |
I-PinfSSR8 | 4 | 250-275 | I-FAM-AATCTGATCGCAACTGAGGG R: GTTTACAAGATACACACGTCGCTCC |
I-PinfSSR4 | 7 | 280-305 | FAM-TCTTGTTCGAGTATGGCGACG R: GTTTCACTTCGGGAGAAAGGCTTC |
UPi04 | 4 | 160-175 | F: VIC-AGCGGCTTACCGATGG R: GTTTCAGCGGCTGTTTTTCGAC |
UPi70 | 3 | 185-205 | F: VIC-ATGAAAATACGTCAATGCTCG R: CGTTGGATATTTCTATTTCTTCG |
I-PinfSSR6 | 3 | 230-250 | F: GTTTTGGTGGGGCTGAAGTTTT R: VIC-TCGCCACAAGATTTATTCCG |
UPi63 | 3 | 265-280 | F: VIC-ATGACGAAGATGAAAGTGAGG R: CGTATTTTCCTGTTTATCTAACACC |
I-PinfSSR2 | 3 | 165-180 | F: PET-CGACTTCTACATCACCGGC R: GTTTGCTTGGACTGCGTCTTTAGC |
I-Pi4B | 5 | 200-295 | F: PET-AAAATAAAGCCTTTGGTTCA R: GCAAGCGAGGTTTGTAGATT |
Isibonelo sokubona ngeso lengqondo imiphumela yokuhlaziywa kuboniswe ku-Fig. 6. Imiphumela yahlaziywa kusetshenziswa isofthiwe ye-GeneMapper 3.7 ngokuqhathanisa idatha etholiwe naleyo yezindawo ezingazodwa ezaziwayo. Ukuze kube lula ukutolikwa kwemiphumela yokuhlaziya, kuyadingeka ukufaka izithenjwa eziyi-1-2 ezihlukanisiwe ezine-genotype eyaziwayo ocwaningweni ngalunye.
Indlela yocwaningo ehlongozwayo yahlolwa enanini elibalulekile lamasampula enkambu, okwathi ngemva kwalokho ababhali bahlanganisa izivumelwano phakathi kwamalabhorethri ezinhlangano ezimbili, iJames Hutton Institute (UK) kanye neWageningen University & Research (Netherlands), okuthi, kanye nethuba lokusebenzisa. amakhadi e-FTA ajwayelekile okuqoqwa lula nokuthunyelwa kwamasampula e-DNA e-P. infestans akwenza kwaba nokwenzeka ukukhuluma mayelana nokwenzeka kokusetshenziswa kwezentengiselwano kwalokhu kuthuthukiswa. Ukwengeza, indlela esheshayo nenembile ye-genotyping P. infestans ihlukanisa kusetshenziswa ukuhlaziya i-multiplex ye-SSR yenze kwaba nokwenzeka ukwenza izifundo ezimisiwe zezibalo zaleli gciwane emhlabeni wonke, nokudalwa kwesizindalwazi somhlaba mayelana nokulimaza kwakamuva ngaphakathi kohlaka lwe-pathogen. Iphrojekthi ye-Eucablight (www.eucablight.org), kuhlanganise , kuhlanganise nemiphumela yokuhlaziywa kwe-microsatellite, yenze kwaba nokwenzeka ukulandelela ukuvela nokusabalalisa kwezinhlobo ezintsha ze-genotypes emhlabeni jikelele.
I-Amplified restriction fragment length polymorphism (AFLP). I-AFLP (i-amplified fragment length polymorphism) iwubuchwepheshe bokukhiqiza izimpawu zamangqamuzana ezingahleliwe kusetshenziswa iziqalo ezithile. Ku-AFLP, i-DNA iphathwa ngenhlanganisela yama-enzyme amabili anomkhawulo. Ama-adaptha athile axhunywe ekugcineni okunamathelayo kwezingcezu zemikhawulo.
Lezi zingcezu zibe sezikhuliswa kusetshenziswa iziqalo ezihambisana nokulandelana kwe-adaptha nesayithi lokukhawulela futhi ngaphezu kwalokho zithwala isisekelo esisodwa noma ngaphezulu ezingahleliwe ekugcineni kwazo ezi-3. Isethi yezingcezu ezitholiwe incike kuma-enzyme akhawulelwe kanye nama-nucleotide akhethwe ngokungahleliwe emaphethelweni angu-3'-emikhawulweni yokuqala (Vos et al., 1995). I-AFLP genotyping isetshenziselwa ukutadisha ngokushesha ukuhlukahluka kofuzo lwezinto ezihlukahlukene eziphilayo.
Incazelo eningiliziwe yendlela inikezwa emisebenzini ka-Mueller, Wolfenbarger, 1999, Savelkoul et al., 1999. Umsebenzi omningi wokuqhathanisa nokuxazululwa kwezindlela ze-AFLP ne-SSR zenziwe abacwaningi baseShayina. Izici ze-phenotypic kanye ne-genotypic ye-48 P. infestans isolate eqoqwe ezifundeni ezinhlanu zaseNyakatho yeChina zacwaningwa. I-spectra ye-AFLP yembula izinhlobo eziyisishiyagalombili ze-DNA genotypes, ngokungafani ne-SSR genotypes, okungekho ukwehluka kwayo okuveziwe (Guo et al., 2008).
Ukukhulisa ngeziqalo ezihambisana nokulandelana kwezinto zeselula
Omaka abasuselwe ekulandeleni kwama-retrotransposons alungele kakhulu ukudwetshwa kwemephu yezofuzo, ucwaningo lokuhlukahluka kofuzo kanye nezinqubo zokuziphendukela kwemvelo (Schulman, 2006). Uma kwenziwa ama-primers ahambisana nokulandelana okuzinzile kwezakhi ezithile zeselula, kungenzeka ukukhulisa izifunda ze-genome ezitholakala phakathi kwazo. Ocwaningweni lwe-ejenti eyimbangela ye-blight sekwephuzile, indlela yokukhulisa izingxenye ze-genome kusetshenziswa i-primer ehambisanayo nokulandelana okuyisisekelo kwe-SINE (I-Short Interspersed Nuclear Elements) retropazone isetshenziswe ngempumelelo (Lavrova no-Elansky, 2003). Le ndlela yembula umehluko ngisho nasenzalweni engashadile yodwa. Mayelana nalokhu, kwafinyelelwa esiphethweni sokuthi indlela ye-inter - SINE - PCR icace kakhulu futhi isivinini sokunyakaza kwezakhi ze-SINE ku-Phytophthora genome siphezulu.
Ku-genome ye-P. infestans, imindeni ye-12 ye-retrotransposons emfushane (SINEs) ikhonjwe; ukusatshalaliswa kwezinhlobo zama-retrotransposon amafushane kwaphenywa; izakhi (SINEs) zatholwa ezitholakala ku-genome ye-P. infestans kuphela (Lavrova, 2004).
Izici zokusetshenziswa kwezindlela zocwaningo lokuqhathanisa lwezinhlobo ezifundweni zabantu
Lapho uhlela isifundo, kubalulekile ukuqonda ngokucacile imigomo esiyiphishekelayo futhi usebenzise izindlela ezifanele. Ngakho-ke, ezinye izindlela zivumela ukukhiqiza inombolo enkulu yezici zomaka ezizimele, kodwa ngesikhathi esifanayo zinokuphindaphindeka okuphansi futhi zincike kakhulu kuma-reagents asetshenzisiwe, izimo zokusabela, kanye nokungcoliswa kwento efundwayo. Ngakho-ke, esifundweni ngasinye seqembu lezinkinga, kuyadingeka ukusebenzisa izihlukanisi eziningana ezijwayelekile (inkomba), kodwa ngisho nakulokhu, imiphumela yokuhlolwa okuningana inzima kakhulu ukuhlanganisa.
Leli qembu lezindlela lihlanganisa i-RAPD, i-AFLP, i-InterSSR, i-InterSINE PCR. Ngemva kokukhulisa, kutholwa inani elikhulu lezingcezu ze-DNA ezinobukhulu obuhlukahlukene. Kutuswa ukusebenzisa amasu anjalo lapho kudingekile ukuthola umehluko phakathi kwezinhlobo ezihlobene eduze (izingane zomzali, i-wild type-mutants, njll.), noma ezimeni lapho ukuhlaziya okuningiliziwe kwesampula encane kudingeka. Ngakho-ke, indlela ye-AFLP isetshenziswa kabanzi ekudwebeni izakhi zofuzo ze-P. infestans (van der Lee et al., 1997) nasezifundweni ze-intrapopulation (Knapova, Gisi, 2002, Cooke et al, 2003, Flier et al, 2003). Izindlela ezinjalo azifanelekile ukuzisebenzisa lapho udala imininingwane yezinkinga, ngoba cishe akunakwenzeka ukuhlanganisa ukubalwa kwezimali kwemiphumela lapho kwenziwa ukuhlaziya kumalabhorethri ahlukene.
Naphezu kokubonakala kulula nokusheshisa kokubulawa (ukuhlukaniswa kwe-DNA ngaphandle kokuhlanzwa okuhle, ukukhulisa, ukubonwa kwemiphumela), leli qembu lezindlela lidinga ukusetshenziswa kwendlela ekhethekile yokubhala imiphumela: i-distillation ku-polyacrylamide gel ebhalwe ukuthi (radioactive noma luminescent) iziqalisi kanye nokuchayeka okulandelayo ezintweni ezikhanyayo noma ezinemisebe. Ukuthwebula kwejeli okujwayelekile kwe-ethidium bromide agarose ngokuvamile akuzilungele lezi zindlela ngoba inombolo enkulu yezingcezu ze-DNA ezinobukhulu obuhlukahlukene zingahlangana.
Ezinye izindlela, ngokuphambene nalokho, zenza kube nokwenzeka ukukhiqiza inombolo encane yezici ngokuphindaphinda kwazo okuphezulu kakhulu. Leli qembu lihlanganisa ukucwaninga kwe-mitochondrial DNA haplotypes (ama-haplotype amabili kuphela i-Ia ne-IIa aphawulwe eRussia), uhlobo lokukhwelana (iningi lama-isolate ahlukaniswe abe izinhlobo ezimbili: i-A2 ne-A1, i-SF eyazizalayo ayitholakali neze) kanye ne-peptidase isozyme spectra ( i-loci emibili Pep2 ne-Pep1, ehlanganisa ama-isozymes amabili ngayinye) kanye ne-glucose-2-phosphate isomerase (eRussia akukho ukuhlukahluka kwalesi sici, nakuba i-polymorphism ephawulekayo ibonakala kwamanye amazwe omhlaba). Kutuswa ukusebenzisa lezi zici lapho uhlaziya amaqoqo, uhlanganisa imininingwane egciniwe yesifunda neyomhlaba wonke. Endabeni yokuhlaziywa kwama-isozymes nama-haplotypes e-mitochondrial DNA, kungenzeka ukwenza ngaphandle kwezinhlobo ezijwayelekile nhlobo, kuyilapho ukuhlaziya izinhlobo zokukhwelana kudinga ukuhlukaniswa okubili kokuhlola nezinhlobo ezaziwayo zokukhwelana.
Izimo zokusabela nama-reagents angathinta kuphela umehluko womkhiqizo ku-electrophoretogram; ukuvezwa kwama-artifact kulezi zinhlobo zocwaningo mancane amathuba okuba.
Njengamanje, iningi labantu engxenyeni yaseYurophu yaseRussia limelelwa yizinhlobo zazo zombili izinhlobo zokukhwelana (Ithebula 6), phakathi kwazo kukhona okuhlukanisiwe okunezinhlobo ze-Ia ne-IIa ze-mitochondrial DNA (ezinye izinhlobo ze-mtDNA ezitholakala emhlabeni bezingekho. etholakala eRussia ngemva kuka-1993). I-spectra ye-peptidase isozymes imelelwa ama-genotypes amabili ku-Pep1 locus (100/100, 92/92 kanye ne-heterozygote 92/100, kanye ne-92/92 genotype ayivelakancane kakhulu (<0,3%)) kanye nama-genotypes amabili ku-Pep 2 locus (100/100 , 112/112 kanye ne-heterozygote 100/112, ne-genotype 112/112 eyenzeka kancane kuno-100/100, kodwa futhi kaningi).
Kwakungekho kuhlukahluka ku-spectrum ye-isozymes ye-glucose-6-phosphate isomerase ngemva kuka-1993 (ukunyamalala komugqa we-clonal US-1); zonke izindawo ezizimele ezihlolisisiwe zine-genotype engu-100/100 (Elansky kanye ne-Smirnov, 2002).
Iqembu lesithathu lezindlela livumela ukuthola iqembu elanele lezici zomaka ezizimele ezinokukhiqizwa kabusha okuphezulu. Namuhla, leli qembu lihlanganisa i-RFLP-RG57 probe, ekhiqiza izingcezu ze-DNA ezingama-25-29 ezinosayizi abahlukene. I-RFLP-RG57 ingasetshenziswa kokubili ekuhlaziyeni amasampula nasekuhlanganisweni kolwazi. Nokho, le ndlela ibiza kakhulu kuneyangaphambili, idla isikhathi, futhi idinga inani elikhulu ngokwanele le-DNA ehlanzwe kakhulu. Ngakho-ke, umcwaningi uphoqeleka ukuthi anciphise umthamo wezinto ezihloliwe.
Ukuthuthukiswa kwe-RFLP-RG57 ekuqaleni kwawo-90s ekhulwini elidlule leminyaka kuqinise kakhulu izifundo zabantu ze-causative agent ye-blight sekwephuzile. Yaba yisisekelo sendlela esekelwe ekukhethweni nasekuhlaziyweni kwe "Clonal lines" (bheka ngezansi). Kanye ne-RFLP-RG57, uhlobo lokuhlangana, i-DNA fingerprinting (indlela ye-RFLP-RG57), i-spectra ye-peptidase ne-glucose-6-phosphate isomerase isoenzymes, kanye nohlobo lwe-DNA ye-mitochondrial kusetshenziswa ukukhomba imigqa ye-clonal. Ngenxa yakhe, kukhonjisiwe al., 1994), ukushintshwa kwabantu abadala ngabasha (Drenth et al, 1993, Sujkowski et al, 1994, Goodwin et al, 1995a), kwembula imigqa yama-clonal ekhona emazweni amaningi umhlaba. Izifundo zezinhlobo zaseRussia zisebenzisa le ndlela zibonise i-polymorphism ephezulu ye-genotypic yezinhlobo zengxenye yaseYurophu kanye ne-monomorphism yezibalo zezingxenye zase-Asia naseMpumalanga Ekude yaseRussia (Elansky et al, 2001). Futhi manje le ndlela iseyiyona eyinhloko ezifundweni zabantu be-P. infestans. Kodwa-ke, ukusatshalaliswa kwayo okubanzi kuphazanyiswa yizindleko zayo eziphakeme kanye nomfutho womsebenzi owenziwayo.
Enye indlela ethembisayo engavamile ukusetshenziswa ocwaningweni lwe-P. infestans ukuhlaziywa kwe-microsatellite repeat (SSR). Njengamanje, le ndlela isetshenziswa kabanzi ukuhlukanisa imigqa yama-clonal. Ukuze kuhlaziywe izinhlobo, izici zomaka ze-phenotypic ezifana nokuba khona kwezakhi zofuzo zezinhlobo zamazambane (Avdey, 1995, Ivanyuk et al., 2002, Ulanova et al., 2003) kanye notamatisi zazisetshenziswa kabanzi (futhi ziyaqhubeka nokusetshenziswa) . Njengamanje, izakhi zofuzo eziyingozi ezinhlotsheni zamazambane sezilahlekelwe inani lazo njengezimpawu zomaka ezifundweni zabantu ngenxa yokuvela kwenani eliphezulu (noma eliseduze nalo) lezakhi zofuzo eziningi ezihlukanisiwe. Ngasikhathi sinye, isakhi sofuzo se-T1 se-virulence sezinhlobo zikatamatisi eziphethe isakhi sofuzo se-Ph1 esihambisanayo sisasetshenziswa ngempumelelo njengesici somaka (Lavrova et al., 2003; Ulanova et al., 2003).
Emisebenzini eminingi, ukumelana nama-fungicides kusetshenziswa njengesici somaka. Lesi sici asithandeki ukuthi sisetshenziswe ezifundweni zenani labantu ngenxa yokubukeka okulula kokuguquka kokumelana emigqeni ye-clonal ngemva kokusetshenziswa kwe-metatalaxyl (noma i-mefenoxam) equkethe izibulala-fungi ensimini. Isibonelo, umehluko omkhulu ezingeni lokumelana ubonisiwe ngaphakathi komugqa we-Sib1 clonal (Elansky et al., 2001).
Ngakho, uhlobo lokukhwelana, i-peptidase isozyme spectrum, uhlobo lwe-DNA ye-mitochondrial, i-RFLP-RG57, i-SSR yizici zomaka ezikhethwayo zokudala amabhange edatha nokulebula izinhlobo zamaqoqo. Ukuze uqhathanise amasampuli alinganiselwe, uma kudingekile ukusebenzisa inombolo enkulu yezici zomaka, ungasebenzisa i-AFLP, RAPD, InterSSR, Inter-SINE PCR (Ithebula 5). Kodwa-ke, kufanele kukhunjulwe ukuthi lezi zindlela azikwazi ukukhiqizwa kabusha, futhi ekuhloleni ngakunye komuntu ngamunye (umjikelezo we-amplification electrophoresis) kuyadingeka ukusebenzisa izinkomba ezimbalwa zokuhlukanisa.
Ithebula 5. Ukuqhathaniswa kwezindlela ezahlukene zocwaningo lwezinhlobo P. infestans
Umbandela | TS | Amaphoyisa ase-Isofer | I-MtDNA | I-RFLP- RG57 | I-RAPD | ISSR | I-SSR | I-AFLP | Rev |
---|---|---|---|---|---|---|---|---|---|
Inani lolwazi | Н | Н | Н | С | В | В | С | В | В |
Ukukhiqiza kabusha | В | В | В | В | Н | Н | С | С | С |
Amathuba wezinto zobuciko | Н | Н | Н | Н | В | С | Н | С | В |
izindleko | Н | С | Н | В | Н | Н | Н | С | Н |
Amandla omsebenzi | Н | Н | Н | В | NS* | NS* | Н | С | NS* |
Isivinini sokuhlaziya ** | В | Н | Н | С | Н | Н | Н | Н | Н |
Qaphela: H - low, C - medium, B - high; НС * - amandla okusebenza aphansi uma usebenzisa ijeli ye-agarose noma okuzenzakalelayo
i-genotyper, medium - by distillation kujeli ye-polyacrylamide enama-primer anelebuli,
** - singabali isikhathi esichithwa ekukhuleni kwe-mycelium yokuhlukaniswa kwe-DNA.
Isakhiwo sabantu
Imigqa yama-Clonal
Uma kungekho ukuhlanganisa kabusha noma ngokunikela kwakho okungatheni esakhiweni sabantu, inani labantu liqukethe inani elithile lama-clones, ukushintshana kwezakhi zofuzo phakathi kwazo okuyivelakancane kakhulu.
Kubantu abanjalo, kufundisa kakhulu ukutadisha hhayi amaza ofuzo ngalunye, kodwa imvamisa yezinhlobo ze-genotype ezinomsuka ofanayo (imigqa yama-clonal noma imigqa yama-clonal) futhi ihluke kuphela ekuguquguqukeni kwamaphuzu. Ucwaningo lwabantu lwe-pathogen yokulimaza sekwephuzile kanye nokuhlaziywa kwemigqa ye-clonal kuye kwashesha kakhulu kusukela kwavela indlela ye-RFLP-RG57 ekuqaleni kwawo-90s wekhulunyaka eledlule. Kanye ne-RFLP-RG57, uhlobo lokukhwelana, i-spectra ye-peptidase ne-glucose-6-phosphate isomerase isoenzymes, nohlobo lwe-mitochondrial DNA kusetshenziswa ukukhomba imigqa ye-clonal. Izici zemigqa evame kakhulu yama-clonal ziboniswa kuThebula lesi-6.
I-Clone US-1 yayibusa abantu yonke indawo kwaze kwaba sekupheleni kweminyaka yawo-80, okwathi ngemva kwalokho yaqala ukuthathelwa indawo amanye ama-clones futhi yanyamalala eYurophu naseNyakatho Melika. Manje isitholakala eMpumalanga Ekude (Philippines, Taiwan, China, Japan, Korea, Koh et al., 1994, Mosa et al, 1993), e-Afrika (Uganda, Kenya, Rwanda, Goodwin et al, 1994, Vega- Sanchez et al., 2000; Ochwo et al., 2002) kanye naseNingizimu Melika (Ecuador, Brazil, Peru, Forbes et al., 1997, Goodwin et al., 1994). Azikho izinhlobo zomugqa we-US-1 ezihlonziwe e-Australia kuphela. Ngokusobala, ama-P. infestans isolate afika e-Australia negagasi elihlukile lokufuduka (Goodwin, 1997).
I-Clone US-6 yasuka enyakatho yeMexico yaya eCalifornia ngasekupheleni kweminyaka yama-70s futhi yabangela ubhubhane lapho kumazambane notamatisi ngemuva kweminyaka engama-32 ingenazo izifo. Ngenxa yolaka lwayo oluphezulu, yasusa i-US-1 clone futhi yaqala ukubusa ogwini olusentshonalanga ye-United States (Goodwin et al., 1995a).
I-genotypes US-7 kanye ne-US-8 yatholwa e-United States ngo-1992 futhi kakade ngo-1994 yasakazwa kabanzi e-United States naseCanada. Ngesizini eyodwa yensimu, i-clone US-8 iyakwazi ukucishe isuse ngokuphelele i-US-1 ezigabeni zamazambane ekuqaleni ezingenwe yiwo womabili ama-clone ngokuhlangana okulinganayo (Miller noJohnson, 2000).
Ama-Clones BC-1 kuya ku-BC-4 akhonjwe e-British Columbia ngenani elincane lezindawo ezingazodwa ukusuka ku-Goodwin et al., 1995b). I-Clone US-11 yasabalala kabanzi e-United States futhi yathatha indawo ye-US-1 e-Taiwan. Ama-Clones JP-1 kanye ne-EC-1, kanye ne-clone US-1, ajwayelekile e-Japan nase-Ecuador, ngokulandelana (Koh et al., 1994; Forbes et al., 1997).
I-SIB-1 iyi-clone eyanqoba eRussia endaweni enkulu ukusuka esifundeni saseMoscow kuya eSakhalin. Esifundeni sase-Moscow, kwatholakala ngo-1993, futhi ezinye zezindawo zazihlanganisa ikakhulukazi izinhlobo zalo mzila we-clonal, amelana kakhulu ne-metaxyl. Ngemuva kuka-1993, ukusabalala kwale clone kwehle kakhulu. Ngaphandle kwe-Urals, ngo-1997-1998, i-SIB-1 yatholakala yonke indawo, ngaphandle kwe-Khabarovsk Territory (i-SIB-2 clone isakazeke lapho). Ukuhlukaniswa kwendawo yama-clones anezinhlobo ezahlukene zokukhwelana akufaki inqubo yocansi eSiberia naseMpumalanga Ekude. Esifundeni saseMoscow, ngokungafani neSiberia, inani labantu limelelwa ama-clones amaningi; cishe yonke i-isolate ine-multilocus genotype eyingqayizivele (Elansky et al., 2001, 2015). Lokhu kwehlukahlukana akukwazi ukuchazwa kuphela ngokungeniswa kwamanye amazwe kwezinhlobo zesikhunta ezivela ezingxenyeni ezihlukene zomhlaba ezinembewu yembewu engenisiwe. Njengoba zombili izinhlobo zokukhwelana zenzeka kubantu, kungenzeka ukuthi ukuhlukahluka kwakho kungenxa yokuhlanganiswa kabusha. Ngakho-ke, eBritish Columbia, ukuvela kwe-genotypes BC-2, BC-3, ne-BC-4 kucatshangwa ngenxa yokuhlanganiswa kwama-clones BC-1 kanye ne-US-6 (Goodwin et al., 1995b). Kungenzeka ukuthi izinhlobonhlobo ze-hybrid nazo zitholakala kubantu baseMoscow. Isibonelo, izinhlobo ze-MO-4, MO-8 kanye ne-MO-11 i-heterozygous ye-PEP locus ingaba ingxube phakathi kwezinhlobo ze-MO-12, MO-21, MO-22, ezinohlobo lokumatanisa lwe-A2 kanye ne-homozygous ye-allele eyodwa ye-PEP. i-locus kanye nohlobo lwe-MO-8, olunohlobo lokukhwelana lwe-A1 kanye ne-homozygous kwenye i-allele ye-locus. Futhi uma lokhu kunjalo, futhi kubantu banamuhla be-P. infestans kukhona ukuthambekela ekwandeni kwendima yenqubo yocansi, khona-ke inani lolwazi lokuhlaziywa kwama-clones we-multilocus lizokwehla (Elansky et al., 2001, 2015) ).
Ukwehluka kwemigqa yama-clonal
Kuze kube ngama-90s wekhulu lama-20, umugqa we-clonal US-1 wawusabalala emhlabeni. Iningi labantu basensimini kanye nesifunda lalihlanganisa kuphela izinhlobo ezine-US-1 genotype. Kodwa-ke, umehluko phakathi kwama-isolate nawo wabonwa, okungenzeka ukuthi ubangelwa inqubo yokuguqula. Uguquko lwenzekile kukho kokubili i-nuclear kanye ne-mitochondrial DNA futhi kwathinteka, phakathi kwezinye izinto, izinga lokumelana nezidakamizwa ze-phenylamide kanye nenani lezakhi zofuzo ze-virulence. Imigqa ehlukile kuma-genotypes asekuqaleni ngokuguqulwa kwezakhi kukhonjiswa ngezinombolo ezengeziwe ngemva kwechashazi elilandela igama le-genotype yoqobo (isibonelo, umugqa oguquguqukayo wase-US-1.1 womugqa we-clonal wase-US-1). Imigqa ye-DNA yezigxivizo zeminwe US-1.5 kanye ne-US-1.6 iqukethe imigqa yesisekeli enosayizi abahlukene (Goodwin et al., 1995a, 1995b); umugqa we-clonal US-6.3 nawo uhlukile ku-US-6 emugqeni owodwa wesisekeli (Goodwin, 1997, Table 7).
Ucwaningo lwe-DNA ye-mitochondrial lwembula ukuthi uhlobo lwe-1b kuphela lwe-DNA ye-mitochondrial olutholakala kumugqa we-clonal US-1 (Carter et al., 1990). Kodwa-ke, ocwaningweni lwezinhlobo zalolu hlu lozalo lwe-clonal oluvela ePeru nasePhilippines, kwatholakala izindawo ezingazodwa izinhlobo zazo ze-DNA ye-mitochondrial zihluke ku-1b ngokuba khona kokufakwayo nokususwa (Goodwin, 1991, Koh et al., 1994).
Ithebula 6. I-Multilocus genotypes yeminye imigqa ye-P. infestans clonal
Isihloko | Uhlobo lokukhwelana | Ama-Isozymes | Izigxivizo zeminwe ze-DNA | Uhlobo lwe-MtDNA | |
I-GPI | PEP | ||||
I-US-1 | I-А1 | 86/100 | 92/100 | I-1.0111010110011E + 24 | Ib |
I-US-2 | I-А1 | 86/100 | 92/100 | I-1.0111010010011E + 24 | - |
I-US-3 | I-А1 | 86/100 | 92/100 | I-1.0111000000011E + 24 | - |
I-US-4 | I-А1 | 100/100 | 92/92 | I-1.0111010010011E + 24 | - |
I-US-5 | A1 | 100/100 | 92/100 | I-1.0111010010011E + 24 | - |
I-US-6 | A1 | 100/100 | 92/100 | I-1.0111110010011E + 24 | IIb |
I-US-7 | A2 | 100/111 | 100/100 | I-1.0011000010011E + 24 | Ia |
I-US-8 | A2 | 100/111/122 | 100/100 | I-1.0011000010011E + 24 | Ia |
I-US-9 | A1 | 100/100 | 83/100 | * | - |
I-US-10 | A2 | 111/122 | 100/100 | - | - |
I-US-11 | A1 | 100/111 | 92/100 | I-1.0101110010011E + 24 | IIb |
I-US-12 | A1 | 100/111 | 92/100 | I-1.0001000010011E + 24 | - |
I-US-14 | A2 | 100/122 | 100/100 | I-1.0000000000011E + 24 | - |
I-US-15 | A2 | 100/100 | 92/100 | I-1.0001000010011E + 24 | Ia |
I-US-16 | A1 | 100/111 | 100/100 | I-1.0001100010011E + 24 | - |
I-US-17 | A1 | 100/122 | 100/100 | I-1.0100010000011E + 24 | - |
I-US-18 | A2 | 100/100 | 92/100 | I-1.0001000010011E + 24 | Ia |
I-US-19 | A2 | 100/100 | 92/100 | I-1.0101010000011E + 24 | Ia |
EC-1 | A1 | 90/100 | 96/100 | I-1.1111010010011E + 24 | I-IIa |
I-SIB-1 | A1 | 100/100 | 100/100 | I-1.0001000110011E + 24 | I-IIa |
I-SIB-2 | A2 | 100/100 | 100/100 | I-1.0001000010011E + 24 | I-IIa |
I-SIB-3 | A1 | 100/100 | 100/100 | I-1.1001010100011E + 24 | I-IIa |
I-MO-1 | A2 | 100/100 | 100/100 | I-1.0001000110011E + 24 | I-IIa |
I-MO-2 | A2 | 100/100 | 100/100 | I-1.0001000010011E + 24 | Ia |
I-MO-3 | A1 | 100/100 | 100/100 | I-1.0101000010011E + 24 | I-IIa |
I-MO-4 | A1 | 100/100 | 92/100 | I-1.0101110110011E + 24 | I-IIa |
I-MO-5 | A1 | 100/100 | 100/100 | I-1.0001010010011E + 24 | I-IIa |
I-MO-6 | A1 | 100/100 | 100/100 | I-1.0101010010011E + 24 | Ia |
I-MO-7 | A1 | 100/100 | 92/100 | I-1.0001000110011E + 24 | I-IIa |
I-MO-8 | A1 | 100/100 | 92/92 | I-1.0101100010011E + 24 | I-IIa |
I-MO-9 | A1 | 100/100 | 92/100 | I-1.0001000010011E + 24 | I-IIa |
I-MO-10 | A1 | 100/100 | 100/100 | I-1.0101100000011E + 24 | Ia |
I-MO-11 | A1 | 100/100 | 92/100 | I-1.0101010010011E + 24 | Ia |
I-MO-12 | A2 | 100/100 | 100/100 | I-1.0101010010011E + 24 | Ia |
I-MO-13 | A1 | 100/100 | 100/100 | I-1.0101010000011E + 24 | Ia |
I-MO-14 | A1 | 100/100 | 100/100 | I-1.01010010011E + 22 | Ia |
I-MO-15 | A1 | 100/100 | 100/100 | I-1.101110010011E + 23 | Ia |
I-MO-16 | A1 | 100/100 | 100/100 | I-1.0001000000011E + 24 | I-IIa |
I-MO-17 | A1 | 86/100 | 100/100 | I-1.0101010110011E + 24 | Ib |
I-MO-18 | A1 | 100/100 | 100/100 | I-1.0101110010011E + 24 | I-IIa |
I-MO-19 | A1 | 100/100 | 100/100 | I-1.0101010000011E + 24 | I-IIa |
I-MO-20 | A2 | 100/100 | 100/100 | I-1.0101010000011E + 24 | I-IIa |
I-MO-21 | A2 | 100/100 | 100/100 | I-1.0101010000011E + 24 | I-IIa |
Qaphela: * - ayikho idatha.
Ithebula 7. I-Multilocus genotypes nemigqa yazo eguquguqukayo
Isihloko | Uhlobo lokukhwelana | | I-DNA Fingerprints (RG57) | Amanothi | |
I-GPI | I-PEP-1 | ||||
I-US-1 | I-А1 | 86/100 | 92/100 | 1011101011001101000110011 | I-genotype yokuqala 1 |
I-US-1.1 | A1 | 86/100 | 100/100 | 1011101011001101000110011 | Ukuguqulwa kwe-PEP |
I-US-1.2 | A1 | 86/100 | 92/100 | 1011101010001101000110011 | Ukushintsha kwe-RG57 |
I-US-1.3 | A1 | 86/100 | 92/100 | 1011101001001101000110011 | Ukushintsha kwe-RG57 |
I-US-1.4 | A1 | 86/100 | 100/100 | 1011101010001101000110011 | Ukuguqulwa kwe-RG57 kanye ne-PEP |
I-US-1.5 | A1 | 86/100 | 92/100 | 1011101011001101010110011 | Ukushintsha kwe-RG57 |
I-US-6 | A1 | 100/100 | 92/100 | 1011111001001100010110011 | I-genotype yokuqala 2 |
I-US-6.1 | A1 | 100/100 | 92 /92 | 1011111001001100010110011 | Ukuguqulwa kwe-PEP |
I-US-6.2 | A1 | 100/100 | 92/100 | 1011101001001100010110011 | Ukushintsha kwe-RG57 |
I-US-6.3 | A1 | 100/100 | 92/100 | 1011111001011100010110011 | Ukushintsha kwe-RG57 |
I-US-6.4 | A1 | 100/100 | 100/100 | 1011011001001100010110011 | Ukuguqulwa kwe-RG57 kanye ne-PEP |
I-US-6.5 | A1 | 100/100 | 92/100 | 1011111001001100010010011 | Ukushintsha kwe-RG57 |
I-BR-1 | A2 | 100/100 | 100/100 | 1011101000001100001111011 | I-genotype yokuqala 3 |
I-BR-1.1 | A2 | 100/100 | 100/100 | 1010101000001100001110011 | Ukushintsha kwe-RG57 |
Kukhona futhi izinguquko ku-spectra ye-isozymes. Njengomthetho, zibangelwa ukuwohloka kwento ephilayo ekuqaleni i-heterozygous yale enzyme ibe yi-homozygous. Ngo-1993, ezithelweni zikatamatisi, sahlonza uhlobo olunezici ze-US-1: iminwe ye-RG57, uhlobo lwe-DNA ye-mitochondrial kanye ne-86/100 genotype ye-glucose-6-phosphati-isomerase, kodwa kwakuyi-homozygous (100/100) i-peptidase locus yokuqala esikhundleni se-92/100 heterozygote ejwayelekile yalo mugqa we-clonal. Siqambe i-genotype yalolu hlobo lwe-MO-17 (Ithebula 6). Imigqa eguquguqukayo i-US-1.1 kanye ne-US-1.4 nayo ihlukile ku-US-1 ngokuguqulwa kwe-peptidase locus yokuqala (Ithebula 7).
Uguquko oluholela ekushintsheni kwenani lezakhi zofuzo ze-virulence zezinhlobo zamazambane notamatisi zivame kakhulu. Baphawulwe phakathi kweziqhelile zomugqa we-clonal US-1 kubantu abavela eNetherlands (Drenth et al., 1994), Peru (Goodwin et al., 1995a), Poland (Sujkowski et al., 1991), enyakatho yeNyakatho Melika ( Goodwin et al., ., 1995b). Umehluko enanini lezakhi zofuzo ze-amazambane virulence waphinde waphawulwa phakathi kwemigqa ye-clonal US-7 kanye ne-US-8 eCanada nase-United States (Goodwin et al., 1995a), phakathi kweziqhelile zomugqa we-SIB-1 e-Asian. ingxenye yeRussia (Elansky et al, 2001).
Ama-Isolates anomehluko oqinile emazingeni okumelana nezidakamizwa ze-phenylamide ahlonzwe emiphakathini yasensimini ye-monoclonal, konke okwakungokwe-clonal line Sib-1 (Elansky et al, 2001, Ithebula 1). Cishe zonke izinhlobo zomugqa we-clonal i-US-1 zingenwa kalula yi-metataxyl; nokho, ama-isolate amelana kakhulu nalo mugqa ahlukaniswa e-Philippines (Koh et al., 1994) nase-Ireland (Goodwin et al., 1996).
Izibalo zesimanje ze-P. infestans
I-Central America (Mexico)
Inani lama-P. infestans e-Mexico lihluke ngokuphawulekayo kwezinye izizwe zomhlaba, okubangelwa ngokuyinhloko indawo yalo yomlando. Ucwaningo oluningi lwalesi sibalo kanye nezinhlobo ezihlobene ze-P. infestans ze-clade Phytophthora, kanye nezinhlobo zendawo zohlobo lwe-Solanum, luholele esiphethweni sokuthi ukuvela kwe-pathogen engxenyeni emaphakathi yeMexico kwenzeka kanye nokuvela kwe-host host. izitshalo futhi yayihlotshaniswa nokuhlanganiswa kocansi (Grünwald, Flier, 2005). Zombili izinhlobo zokukhwelana zimelelwa enanini labantu, futhi ngezilinganiso ezilinganayo, kanye nokuba khona kwama-oospores enhlabathini, ezitshalweni nezilimo eziyizigaxa zamazambane kanye nezinhlobo ezihlobene ezikhula zasendle I-Solanum iqinisekisa ukuba khona kwenqubo yocansi kubantu (i-Fernández- Pavía et al., 2002). Ucwaningo lwakamuva lweSigodi SaseToluca nezindawo ezisizungezile (isikhungo sokucabangela semvelaphi ye-pathogen) luqinisekise ukwehlukahlukana okuphezulu kwezakhi zofuzo zabantu bendawo be-P. infestans (134 multilocus genotypes kusampula yamasampula angu-176) kanye nokuba khona kwama-subpopulations ambalwa ahlukene. esifundeni (Wang et al., 2017). Izinto ezinomthelela kulo mehluko ukuhlukaniswa kwezindawo ezihlala ezindaweni eziphakeme ezimaphakathi neMexico, umehluko ezimeni zokulinywa nezinhlobonhlobo zamazambane ezisetshenziswa ezigodini nasezintabeni, kanye nokuba khona kwezinhlobo ze-Solanum zasendle ezinama-tuberous ezingasebenza njengamanye abamba (Fry et al. ., 2009).
Kodwa-ke, kufanele kuqashelwe ukuthi inani labantu base-P. infestans enyakatho ye-Mexico linemvelo kakhulu futhi lifana kakhulu nenani labantu baseNyakatho Melika, okungase kubonise ukuthi lezi yizinhlobo ezintsha ze-genotypes (Fry et al., 2009).
ENyakatho Melika
Izibalo zaseNyakatho Melika ze-P. infestans bezilokhu zinesakhiwo esilula kakhulu futhi uhlamvu lwazo lwe-clonal lwasungulwa kudala ngaphambi kokusetshenziswa kokuhlaziywa kwe-microsatellite. Kuze kube ngu-1987, umugqa we-clonal US-1 ubungamele e-United States naseCanada (Goodwin et al., 1995). Maphakathi nawo-70s, lapho kuvela ama-fungicides asuselwa ku-metataxyl, le clone yaqala ukuthathelwa indawo amanye, ama-genotypes amelana kakhulu asuka eMexico (Goodwin et al., 1998). Ekupheleni kweminyaka yama-90s. i-genotype yase-US-8 ithathele indawo ngokuphelele i-US-1 genotype e-United States futhi yaba umugqa we-clonal ovelele kumazambane (Fry et al., 2009; Fry et al., 2015). Kuthuthukiswe isimo esihlukile ngotamatisi, lapho imigqa eminingana yama-clonal yayihlala ikhona, futhi ukwakheka kwawo kwashintsha unyaka nonyaka (Fry et al., 2009).
Ngo-2009, kwaqubuka ubhubhane olukhulu lwe-blight ephuzile e-United States kumatamatisi. Isici salolu bhubhane kwaba ukuqala kwawo cishe ngasikhathi sinye ezindaweni eziningi enyakatho-mpumalanga ye-United States, futhi kwahlotshaniswa nokuthengiswa okukhulu kwezithombo zikatamatisi ezinegciwane ezikhungweni zengadi ezinkulu (Fry et al., 2013). Ukulahlekelwa kwezitshalo bekukukhulu. Ukuhlaziywa kwe-Microsatellite yamasampula athintekile kwembula ukuthi uhlobo lobhubhane lungokwe-clonal line US-22 A2-type mating. Ngo-2009, isabelo sale genotype kubantu baseMelika be-P. infestans safinyelela ku-80% (Fry et al., 2013). Eminyakeni eyalandela, ingxenye ye-genotypes enolaka i-US-23 (ikakhulukazi kutamatisi) kanye ne-US-24 (kumazambane) yanda kancane kubantu, nokho, ngemuva kuka-2011, izinga lokutholwa kwe-US-24 lehle kakhulu, futhi kuze kube manje, cishe u-90% wabantu be-pathogen e-United States imelwe yi-US-23 genotype (Fry et al., 2015).
ECanada, njengase-United States, ekupheleni kweminyaka yama-90s. i-genotype evelele i-US-1 yathathelwa indawo yi-US-8, izikhundla ezivelele ezihlale zingashintshile kwaze kwaba ngu-2008. E-Canada, kube nezifo eziwumshayabhuqe ezibi kakhulu ezihambisana nokudayiswa kwezithombo zikatamatisi ezithelelekile, kodwa zabangelwa i-genotypes US-2009 kanye ne-US-2010 (Kalischuk et al., 23). Ukuhlukaniswa okucacile kwezindawo zalezi zinhlobo ze-genotype kwakumangalisa: i-US-8 yayibusa izifundazwe ezisentshonalanga yeCanada (2012%), kuyilapho i-US-23 yayibusa izifundazwe ezisempumalanga (68%). Eminyakeni eyalandela, i-US-8 yasakazekela ezifundeni ezisempumalanga; nokho, ngokuvamile, isabelo sayo kubantu sehla kancane uma kuqhathaniswa nesizinda sokuvela kwe-US-83 kanye ne-US-23 genotypes ezweni (Peters et al., 22). Kuze kube manje, i-US-24 igcina isikhundla esiphezulu kulo lonke elaseCanada; I-US-2014 ikhona e-British Columbia, kuyilapho i-US-23 ne-US-8 zikhona e-Ontario (Peters, 23).
Ngakho, inani labantu baseNyakatho Melika be-P. infestans ikakhulukazi imigqa yama-clonal. Eminyakeni engu-40 edlule, inani le-clonal genotypes elitholiwe selifinyelele ku-24. Naphezu kweqiniso lokuthi izinhlobo zazo zombili izinhlobo zokukhwelana zikhona kubantu, amathuba okuvela kwezinhlobo ezintsha ze-genotype ngenxa yokuhlangana kabusha kocansi ahlala ephansi kakhulu. Noma kunjalo, eminyakeni engu-20 edlule, amacala amaningana okuvela kwe-ephemeral recombinant recombinant aqoshiwe (Gavino et al., 2000; Danies et al., 2014; Peters et al., 2014), futhi esimweni esisodwa, umphumela. Ukuwela kwakuyi-genotype US-11, egxile eNyakatho Melika iminyaka eminingi (Gavino et al., 2000). Kuze kube ngu-2009, izinguquko esakhiweni sezibalo zazihlotshaniswa nokuvela kwezinhlobo ze-genotypes ezintsha, ezinolaka ngokufuduka kwazo okwalandela kanye nokugudluzwa kwabanduleli ababebusa phambilini. Kwenzekani ngo-2009-2010 E-USA naseCanada, i-epiphytotics okokuqala ngqa yabonisa ukuthi ngesikhathi sokuhwebelana kwembulunga yonke, ukuqubuka kwalesi sifo kungahlotshaniswa nokusabalala okusebenzayo kwezinhlobo ezintsha zohlobo lwe-genotype lapho kuthengiswa izinto zokutshala ezinegciwane.
ENingizimu Melika
Kuze kube muva nje, ucwaningo lwezibalo zabantu baseNingizimu Melika be-P. infestans lwalungekho olwejwayelekile noma oluncane. Kuyaziwa ukuthi ukwakheka kwalezi zibalo kulula futhi kufaka phakathi imigqa yama-clonal angu-1-5 ezweni ngalinye (Forbes et al., 1998). Ngakho, ngo-1998, i-genotypes US-1 (Brazil, Chile) BR-1 (Brazil, Bolivia, Uruguay, Paraguay), EC-1 (Ecuador, Colombia, Peru kanye Venezuela), AR-1, AR -2, AR -3, AR-4 kanye ne-AR-5 (Argentina), PE-3 kanye ne-PE-7 (eningizimu yePeru). Uhlobo lokukhwelana u-A2 lwalukhona e-Brazil, Bolivia nase-Argentina futhi alutholakalanga ngale komngcele we-Bolivian-Peruvia endaweni yaseLake Titicaca, ngemva kwalokho i-EC-1 A1 genotype yayigcwele e-Andes. Kutamatisi, i-US-1 yahlala iyi-genotype ehamba phambili kulo lonke elaseNingizimu Melika.
Isimo saqhubeka ngeminyaka yawo-2000s. Iphuzu elibalulekile kwaba ukutholakala komugqa omusha we-clonal EC-2 wohlobo lwe-A2 ezihlotsheni zasendle zamazambane (i-S. brevifolium ne-S. tetrapetalum) eNyakatho ye-Andes (u-Oliva et al., 2010). Ucwaningo lwe-Phylogenetic luye lwabonisa ukuthi lo mugqa awufani ngokuphelele ne-P. infestans, nakuba ihlobene eduze nayo, mayelana nalokho okwakuhlongozwa ukuyicabangela, kanye nomunye umugqa, i-EC-3, ehlukanisiwe nesihlahla sikatamatisi. S. betaceum emila e-Andes, uhlobo olusha olubizwa nge-P. andina; kodwa-ke, isimo salolu hlobo (uhlobo oluzimele noma ingxube ye-P. infestans enomugqa othile ongakaziwa) asikacaci (Delgado et al., 2013).
Njengamanje, zonke izakhamuzi zaseNingizimu Melika ze-P. infestans zingama-clonal. Naphezu kokuba khona kwalezi zinhlobo zombili zokukhwelana, azikho izixuku ezihlonziwe. Kutamatisi, i-genotype yase-US-1 itholakala yonke indawo, ngokusobala isuswe kumazambane ngenxa yobunzima bendawo, umsuka wayo oqondile namanje ongakaziwa. I-BR-1 genotype ikhona eBrazil, eBolivia nase-Uruguay; ePeru, kanye ne-US-1 kanye ne-EC-1, kukhona ezinye izinhlobo ze-genotypes zasendaweni. E-Andes, isikhundla esiphezulu sigcinwa umugqa we-clonal EC-1, ubudlelwano babo ne-P. andina esanda kutholwa buhlala bungaziwa. Indawo kuphela "engazinzile" lapho isikhathi sika-2003-2013. kube nezinguquko ezibalulekile kubantu, kwaba yiChile (Acuña et al., 2012), lapho ngo-2004-2005. inani le-pathogen labonakala ngokumelana ne-metaxyl kanye ne-mitochondrial DNA haplotype entsha (Ia esikhundleni se-Ib ekhona ngaphambilini). 2006 kuya ku-2011 Emphakathini, i-genotype 21 (ngokusho kwe-SSR) yayibusa, isabelo esafinyelela ku-90%, okwathi ngemva kwalokho intende yesandla yadlulela ku-genotype 20, imvamisa yokwenzeka kwayo eminyakeni emibili eyalandela yagcinwa cishe ku-67% (Acuña, 2015).
IYurophu
Emlandweni waseYurophu, kwakukhona okungenani amagagasi amabili okufuduka kwe-P. infestans esuka eNyakatho Melika: ngekhulu le-1. (HERB-1) nasekuqaleni kwekhulu lamashumi amabili (US-70). Ukusatshalaliswa okusabalele kwama-fungicides aqukethe i-metantaxyl ngeminyaka yama-1s. kuholele ekususweni kwe-genotype evelele i-US-XNUMX kanye nokushintshwa kwayo ngama-genotypes amasha. Ngenxa yalokho, emazweni amaningi aseNtshonalanga Yurophu, inani labantu be-pathogen lalimelwe ikakhulukazi imigqa eminingana yama-clonal.
Ukusetshenziswa kohlaziyo lwe-microsatellite ukuze kuhlaziywe inani le-pathogen kwenze kwaba nokwenzeka ukudalula izinguquko ezinkulu ezenzeka eNtshonalanga Yurophu ngo-2005-2008. Ngo-2005, kwatholakala uhlu olusha lwe-clonal e-Great Britain, olubizwa nge-13_A2 (noma "Blue 13" ) futhi ibonakala ngohlobo lwe-A2 lokukhwelana, ulaka oluphezulu nokumelana nama-phenylamide (Shaw et al., 2007). I-genotype efanayo yatholakala kumasampula aqoqwe ngo-2004 eNetherlands nasenyakatho yeFrance, okusikisela ukuthi yathuthela e-UK isuka ezwekazini laseYurophu, ngokunokwenzeka nembewu yamazambane (Cooke et al., 2007). Ucwaningo lwe-genome yabameleli balo mugqa we-clonal lubonise izinga eliphezulu le-polymorphism yokulandelana kwayo (ngowezi-2016, inani lokuhluka kwayo kwe-subclonal lafinyelela ku-340) kanye nezinga elibalulekile lokuhluka kwezinga lokuvezwa kofuzo, kuhlanganisa. izakhi zofuzo ngesikhathi sokutheleleka kwezitshalo (Cooke et al., 2012; Cooke, 2017). Lezi zici, kanye nokwenyuka kwesikhathi kwesigaba se-biotrophic, bekungabangela ukwanda kolaka lwe-13_A2 kanye nekhono layo lokuthelela ngisho nezinhlobo zamazambane ezimelana nokulimaza sekwephuzile.
Eminyakeni embalwa eyalandela, i-genotype yasakazeka ngokushesha kuwo wonke amazwe aseNyakatho-ntshonalanga Yurophu (iGreat Britain, Ireland, France, Belgium, Netherlands, Germany) kanye nokugudluzwa kanyekanye kwezinhlobo ze-genotype ezazibusa ngaphambili 1_A1, 2_A1, 8_A1 (Montarry et al. , 2010; Gisi et al., 2011; Van den Bosch et al., 2011; Cooke, 2015; Cooke, 2017). Ngokusho kwewebhusayithi ethi www.euroblight.net, isabelo se-13_A2 kubantu bala mazwe sifinyelele ku-60-80% nangaphezulu; ukuba khona kwalolu hlobo lofuzo kwaqoshwa nakwamanye amazwe aseMpumalanga naseNingizimu Yurophu. Nokho, ngo-2009-2012. I-13_A2 ilahlekelwe yizikhundla zayo ezivelele e-Great Britain nase-France, ivumela umugqa we-6_A1 (e-Ireland - 8_A1), futhi e-Netherlands nase-Belgium yathathelwa indawo yi-genotypes 1_A1, 6_A1 no-33_A2 (Cooke et al., 2012; Cooke, 2017; Stellingwerf, 2017).
Kuze kube manje, cishe amaphesenti angama-70 abantu baseWestern Europe be-P. infestans bane-monoclonal. Ngokusho kwewebhusayithi www.euroblight.net, i-genotypes evelele emazweni aseNyakatho-ntshonalanga yeYurophu (UK, France,
I-Netherlands, Belgium) isele, cishe ngezilinganiso ezilinganayo, 13_A2 kanye no-6_A1, futhi lokhu kwakamuva akwenzeki ngaphandle kwesifunda esishiwo (ngaphandle kwe-Ireland), kodwa kakade sekunama-subclones okungenani angu-58 (Cooke, 2017). Izinguquko 13_A2 zikhona ngezinombolo eziphawulekayo eJalimane, futhi zibuye zibonwe ngezikhathi ezithile emazweni aseCentral naseNingizimu Yurophu. I-Genotype 1_A1 yenza ingxenye ebalulekile yabantu base-Belgium futhi ingxenye ye-Netherlands ne-France. I-Genotype 8_A1 izinzile kubantu base-Europe ezingeni lika-3-6%, ngaphandle kwase-Ireland, lapho igcina khona isikhundla sayo esiholayo futhi ihlukaniswe yaba ama-subclones amabili (Stellingwerf, 2017). Ekugcineni, ngo-2016, kwaba nokwanda kokuvama kokuvela kwe-genotypes entsha 36_A2 kanye ne-37_A2, okokuqala ebhalwe ngo-2013-2014; kuze kube manje, lezi genotypes zitholakala eNetherlands naseBelgium futhi ngokwengxenye eFrance naseJalimane, kanye naseningizimu yeGreat Britain (Cooke, 2017). Cishe i-20-30% yabantu baseNtshonalanga Yurophu imelelwa i-genotypes eyingqayizivele minyaka yonke.
Ngokungafani neNtshonalanga Yurophu, ngesikhathi kuvela i-genotype ye-13_A2, abantu baseNyakatho Yurophu (iSweden, Norway, Denmark, Finland) babengamelwe yimigqa yama-clonal, kodwa inani elikhulu lama-genotypes ahlukile (Brurberg et al.,
2011). Ngesikhathi sokusabalala okusebenzayo kwe-13_A2 eNtshonalanga Yurophu, ukuba khona kwalesi genotype eScandinavia akuzange kuphawulwe kuze kube ngu-2011, lapho kutholakala okokuqala eNyakatho Jutland (Denmark), lapho ikakhulukazi izinhlobo zamazambane ezimboni zitshalwa ngokusetshenziswa okusebenzayo kwe-metaxyl. -aqukethe ama-fungicides (Nielsen et al., 2014). Ngokusho kwe-www.euroblight.net, i-genotype 13_A2 iphinde yatholwa kumasampula amaningana avela eNorway naseDenmark ngo-2014 kanye namasampula amaningana aseNorway ngo-2016; ngaphezu kwalokho, ngo-2013 eFinland ukuba khona kwenani elincane le-6_A1 genotype kwaphawulwa. Isizathu esiyinhloko sokwehluleka kwe-13_A2 neminye imigqa yama-clonal ekunqotshweni kweScandinavia kubhekwa njengokwehluka kwesimo sezulu salesi sifunda emazweni aseNtshonalanga Yurophu.
Ngaphezu kweqiniso lokuthi ihlobo elipholile nobusika obubandayo bunomthelela ekusindeni kwama-oospores kune-vegetative mycelium (Sjöholm et al., 2013), ukugodola kwenhlabathi ebusika (okungavamile ukwenzeka emazweni afudumele aseNtshonalanga Yurophu) kunomthelela ekubandeni ukuvumelanisa ukuhluma kwe-oospores nokutshala. amazambane, okuthuthukisa indima yawo njengomthombo wokutheleleka okuyinhloko (Brurberg et al., 2011). Kufanele futhi kuqashelwe ukuthi, ezimeni ezisenyakatho, ukuthuthukiswa kokutheleleka okuvela ku-oospores kudlula ukuthuthukiswa kokutheleleka nge-tuberous, okugcina kuvimbele ukubusa kolaka, kodwa kamuva kwathuthukiswa imigqa ye-clonal (Yuen, 2012). Isakhiwo sezibalo ezifundwe kakhulu ze-P. infestans eMpumalanga Yurophu (i-Poland, i-Baltic States) ifana kakhulu neyaseScandinavia.
Zombili izinhlobo zokukhwelana zikhona lapha, futhi iningi lama-genotypes anqunywe ukuhlaziya kwe-SSR ahlukile (Chmielarz et al., 2014; Runno-Paurson et al., 2016). NjengaseNyakatho Yurophu, ukusabalala kwemigqa yama-clonal (ikakhulukazi ye-13_A2 genotype) akubathinti abantu bendawo be-pathogen, egcina izinga eliphezulu lokuhlukahluka ngokungabikho kwemigqa evelele.
Ukuba khona kwe-13_A2 kubonakala ngezikhathi ezithile emasimini anezinhlobo zamazambane ezithengiswayo. ERussia, isimo sithuthuka ngendlela efanayo. Ukuhlaziywa kwe-Microsatellite ye-P. infestans isolate eqoqwe ngo-2008-2011 ezifundeni eziyi-10 ezihlukene zengxenye yaseYurophu yaseRussia, kubonise izinga eliphezulu lokuhlukahluka kwe-genotypic kanye nokuntuleka okuphelele kokuqondana nemigqa yama-clonal yaseYurophu (Statsyuk et al., 2014). Eminyakeni embalwa kamuva, ucwaningo lwamasampula e-P. infestans aqoqwe esifundeni sase-Leningrad ngo-2013-2014 lubonise umehluko omkhulu phakathi kwabo kanye nama-genotypes avela kulesi sifunda ahlonzwe esifundweni sangaphambilini. Kuzo zombili izifundo, ama-genotypes aseNtshonalanga Yurophu awatholakalanga (Beketova et al., 2014; Kuznetsova et al., 2016).
Ukuhlukahluka kofuzo okuphezulu kwabantu baseMpumalanga Yurophu be-P. infestans kanye nokungabikho kwemigqa yama-clonal evelele kuzo kungase kube ngenxa yezizathu ezimbalwa. Okokuqala, njengaseNyakatho Yurophu, izimo zezulu zamazwe acatshangelwayo zifaka isandla ekwakhekeni kwama-oospores njengomthombo oyinhloko wokutheleleka (Ulanova et al., 2010; Chmielarz et al., 2014). Okwesibili, ingxenye enkulu yamazambane akhiqizwa kulawa mazwe atshalwa emapulazini amancane azimele, ngokuvamile azungezwe amahlathi noma ezinye izithiyo ekuhambeni ngokukhululekile kwezinto ezithathelwanayo (Chmielarz et al., 2014). Njengomthetho, amazambane atshalwe ngaphansi kwezimo ezinjalo awaphathwa ngamakhemikhali, futhi ukukhethwa kwezinhlobonhlobo kusekelwe ekuphikisweni kwawo kwe-blight sekwephuzile, i.e. ayikho ingcindezi ekhethiwe yokuba nolaka kanye nokumelana ne-metaxyl, encisha i-genotypes eqinile, njenge-13_A2, yezinzuzo ngaphezu kwamanye ama-genotypes (Chmielarz et al., 2014). Ekugcineni, ngenxa yobuncane bezindawo zomhlaba, abanikazi bazo ngokuvamile abakwenzi ukushintshanisa izitshalo, ukutshala amazambane iminyaka endaweni efanayo, okunomthelela ekuqoqweni kwe-inoculum ehlukahlukene yofuzo (Runno-Paurson et al., 2016; Elansky , 2015; Elansky et al., 2015).
E-Asia
Kuze kube muva nje, ukwakheka kwabantu be-P. infestans e-Asia kusalokhu kungaqondakali kahle. Kwakwaziwa ukuthi imelelwa ikakhulukazi imigqa ye-clonal, futhi umthelela wokuhlangana kabusha kocansi ekuveleni kwama-genotypes amasha mancane kakhulu. Ngakho, isibonelo, ngo-1997-1998. Engxenyeni yase-Asia yaseRussia (eSiberia naseMpumalanga Ekude), inani le-pathogen lalimelwe yizinhlobo ezintathu kuphela ze-genotype ezinobukhulu be-SIB-1 genotype (Elansky et al., 2001). Ukuba khona kwemigqa ye-clonal pathogen kubonisiwe emazweni afana ne-China, Japan, Korea, Philippines kanye ne-Taiwan (Koh et al., 1994; Chen et al., 2009). Ulayini we-clonal US-1 ubungamele endaweni enkulu yase-Asia ngasekupheleni kwawo-90s - ekuqaleni kwawo-2000. cishe yonke indawo yaqala ukuthathelwa indawo ezinye izinhlobo ze-genotype, nazo, zadedela ezintsha. Ezimweni eziningi, izinguquko esakhiweni nasekubunjweni kwabantu emazweni ase-Asia zazihlotshaniswa nokufuduka kwama-genotypes amasha avela ngaphandle. Ngakho-ke, eJapane, ngaphandle kwe-JP-3 genotype, zonke ezinye izinhlobo ze-genotype zaseJapane ezavela ngemva kwe-US-1 (JP-1, JP-2, JP-3) zinemvelaphi yangaphandle efakazelwe ngaphezulu noma engaphansi (Akino et al. , 2011) ... Njengamanje kunezinhlobo ezintathu ze-pathogen eziyinhloko e-China, ezinokuhlukaniswa okucacile kwendawo; ukugeleza kofuzo phakathi kwalezi zibalo akukho noma kubuthakathaka kakhulu (Guo et al., 2010; Li et al., 2013b). I-Genotype 13_A2 yavela endaweni yase-China ezifundazweni zayo eziseningizimu (i-Yunnan ne-Sichuan) ngo-2005-2007, nango-2012-1014. yabuye yabonakala enyakatho-mpumalanga yezwe (Li et al., 2013b). E-India, i-13_A2 yavela cishe ngasikhathi sinye njengase-China, okungenzeka ukuthi inamazambane atheleleke ngembewu (Chowdappa et al., 2015), nango-2009-2010. kubangele i-epiphytosis eyingozi yokulimaza sekwephuzile kutamatisi eningizimu yezwe, okwathi ngemuva kwalokho yasakazekela kumazambane kwathi ngo-2014 yabangela ukuqubuka kwesifo esibi eNtshonalanga Bengal, okuholele ekulimaleni nasekuzibulaleni kwabalimi abaningi bendawo (Fry, 2016) ).
Afrika
Kuze kube ngu-2008-2010. ucwaningo oluhlelekile lwe-P. infestans emazweni ase-Afrika alwenziwanga. Okwamanje, abantu base-Afrika be-P. infestans bangahlukaniswa ngamaqembu amabili, futhi lokhu kuhlukaniswa kuhlotshaniswa ngokucacile neqiniso lokungeniswa kwembewu yamazambane evela eYurophu.
ENyakatho Afrika, engenisa ngenkuthalo amazambane embewu evela e-Europe, uhlobo lokukhwelana lwe-A2 lumelelwe kabanzi cishe kuzo zonke izifunda, okunikeza ithuba lokuvela kwezinhlobo ezintsha ze-genotypes ngenxa yokuhlangana kabusha kwezocansi (Corbière et al., 2010; Rekad et al., 2017). Ngaphezu kwalokho, e-Algeria, ukuba khona kwama-genotypes 13_A2, 2_A1, kanye no-23_A1 anokubusa okukhulu kweyokuqala kuyaphawulwa, kanye nokwehla kancane kancane kwengxenye ye-genotypes ehlukile yokunyamalala ngokuphelele (Rekad et al., 2017) ). Ngokungafani naso sonke isifunda, e-Tunisia (ngaphandle kwenyakatho-mpumalanga yezwe), inani le-pathogen limelelwa ikakhulukazi uhlobo lokukhwelana lwe-A1 (Harbaoui et al., 2014).
Ulayini we-clonal NA-01 ukhonya lapha. Ngokuvamile, ingxenye yemigqa yama-clonal kubantu ingama-43% kuphela. EMpumalanga naseNingizimu Afrika, lapho amanani embewu ethengwayo evela kwamanye amazwe emincane ngokushabalala (Fry et al., 2009), i-P. infestans imelwe imigqa emibili kuphela yohlobo lwe-clonal A1, i-US-1 kanye ne-KE-1, kanye neyokugcina isusa okwakuqala kumazambane (Pule et al., 2012; Njoroge et al., 2016). Kuze kube manje, womabili lawa ma-genotype anenani elibonakalayo lokuhlukahluka kwe-subclonal.
Австралия
Umbiko wokuqala wokulimaza kwamazambane sekwephuzile e-Australia uhlehle ngo-1907, kanti i-epiphytotia yokuqala, okungenzeka ukuthi ibangelwa izimvula ezinamandla ezinyangeni zasehlobo, yenzeka ngo-1909-1911. (UDrenth et al., 2002). Ngokujwayelekile, nokho, ukubola sekwephuzile akunakubaluleka okubalulekile kwezomnotho ezweni. Ukuqubuka okungavamile kokulimaza kwakamuva, okucasulwa izimo zezulu ezinikeza umswakama ophezulu, akwenzeki kaningi ngaphezu kokukodwa njalo eminyakeni engu-5-7 futhi kwenzeka endaweni ikakhulukazi enyakatho yeTasmania nasenkabeni yeVictoria. Ngokuphathelene nalokhu okungenhla, cishe azikho izincwadi ezinikelwe ocwaningweni lwesakhiwo sabantu base-Australia be-P. infestans. Ulwazi lwakamuva olutholakalayo lusukela ku-1998-2000. (UDrenth et al., 2002). Ngokusho kwababhali, inani labantu baseVictoria laliwumugqa we-clonal US-1.3, owaqinisekisa ngokungaqondile ukufuduka kwalesi genotype kusuka e-United States. Izibonelo ze-Tasmanian zahlukaniswa njenge-AU-3, ehlukile kuma-genotypes ayekhona ngaleso sikhathi kwezinye izingxenye zomhlaba.
Izici zokuthuthukiswa kokulimala okwenzeka sekwedlule isikhathi eRussia
EYurophu, ukutheleleka okwethulwa ngezilimo eziyizigaxa zembewu ezigulayo, ama-oospores agcwele ubusika emhlabathini, kanye ne-zoosporangia elethwa umoya ovela ezitshalweni ezitshalwe ezitshalweni ezikhule kakhulu emasimini angonyaka odlule (izitshalo "zamavolontiya"), noma izinqwaba zamabhukhimakhi aqotshiwe ukuze agcinwe. nezilimo eziyizigaxa. Kulezi, izitshalo ezitshalwe ngezinqwaba zezilimo eziyizigaxa ezilahliwe zibhekwa njengomthombo oyingozi kakhulu wokutheleleka. lapho inani lezilimo eziyizigaxa ezihlumayo livame ukuphawuleka, futhi i-zoosporangia ingathwalwa isuka kuzo ngokuhamba amabanga amade. Eminye imithombo (ama-oospores, izitshalo "zamavolontiya") ayiyingozi kangako, ngoba akuyona inkambiso ukutshala izitshalo emasimini afanayo kaningi ngaphezu kokukodwa njalo eminyakeni engu-3-4. Ukutheleleka ngembewu yezilimo eziyizigaxa ezinezifo nakho kuncane ngenxa yohlelo oluhle lokulawula izinga lembewu.
Ngokuvamile, inani le-inoculum kubantu baseYurophu lilinganiselwe, ngakho-ke ukukhula kwalolu bhubhane kuhamba kancane futhi kungalawuleka ngempumelelo kusetshenziswa amalungiselelo amakhemikhali e-fungicidal. Umsebenzi oyinhloko ezimweni zaseYurophu ukulwa nokutheleleka esigabeni lapho ukuhlakazeka okukhulu kwe-zoosporangia kusuka ezitshalweni ezinegciwane kuqala.
E-Russia, isimo sihluke kakhulu. Iningi lezitshalo zamazambane kanye notamatisi litshalwa ezingadini ezincane ezizimele; Izinyathelo zokuzivikela azenziwanga nhlobo kubo, noma ukwelashwa kwe-fungicidal kwenziwa ngenombolo enganele futhi kuqale ngemuva kokuvela kokubola sekwephuzile eziqongweni. Ngenxa yalokho, izingadi zemifino ezizimele zisebenza njengomthombo oyinhloko wokutheleleka, lapho i-zoosporangia ithwalwa khona umoya iye ezitshalweni zentengiso. Lokhu kuqinisekiswa ngokubona kwethu okuqondile ezindaweni zaseMoscow, Bryansk, Kostroma, Ryazan: ukunqotshwa kwezitshalo ezingadini ezizimele kubonakala ngisho nangaphambi kokuqala kokwelashwa kwesikhunta kokutshala kwezentengiselwano. Kamuva, ubhubhane emasimini amakhulu luvinjelwa ukusetshenziswa kwamalungiselelo okubulala isikhunta, kuyilapho ezingadini ezizimele kuba nokuthuthuka okusheshayo kokulimaza sekwephuzile.
Endabeni yokucubungula okungalungile noma "isabelomali" sokutshala kwezentengiselwano, i-foci ye-blight ephuzile ivela emasimini; ngokuhamba kwesikhathi bathuthuka ngenkuthalo, bahlanganisa izindawo ezikhudlwana (Elansky, 2015). Ukutheleleka ezingadini ezizimele kunomthelela omkhulu ezikhungweni eziwubhubhane emikhakheni yezohwebo. Kuzo zonke izifunda ezitshala amazambane zaseRussia, indawo ehlala amazambane ezingadini ezizimele inkulu ngokuphindwe kaningana kunengqikithi yamasimu abakhiqizi abakhulu. Esimweni esinjalo, izingadi zemifino ezizimele zingabhekwa njengensiza ye-inoculum yomhlaba wonke yemikhakha yezohwebo. Ake sizame ukuhlonza lezo zakhiwo eziyisici se-genotypes yezinhlobo ezingadini ezizimele.
Ukutshala amazambane angazange adlulise imbewu nokuhlukaniswa kwabantu, imbewu katamatisi etholwe kubakhiqizi abangabazisayo bangaphandle, ukutshalwa isikhathi eside kwamazambane notamatisi ezindaweni ezifanayo, ukwelashwa okungafanele kwe-fungicide noma ukungabikho kwabo ngokuphelele kuholela kuma-epiphytoties aqinile emkhakheni ozimele, umphumela wawo ukuwela mahhala, ukuxutshwa kanye nokwakheka kwe-oospore ezingadini ezizimele. Ngenxa yalokho, ukuhlukahluka kwe-genotypic okuphezulu kakhulu kwe-pathogen kuyabonwa, lapho cishe lonke uhlobo lwe-genotype luhlukile (Elansky et al., 2001, 2015). Ukutshala imbewu yamazambane anemvelaphi ehlukahlukene yofuzo kwenza kungenzeki ukuthi imigqa ye-clonal ekhethekile yokuhlasela uhlobo oluthile izovela. Izinhlobo ezikhethiwe kuleli cala zihlukahlukene maqondana nezinhlobo ezithintekile, iningi lazo linenani elisondele kakhulu lezakhi zofuzo ze-virulence. Lokhu kuhluke kakhulu ohlelweni "lwemigqa ye-clonal" evamile emikhakheni emikhulu yamabhizinisi ezolimo enohlelo olufakwe kahle lokuvikela ngokumelene nokulimaza sekwephuzile. "Imigqa ye-Clonal" (lapho zonke izinhlobo ze-pathogen ephuzi ephuzile ensimini zimelelwa yi-genotype eyodwa noma eziningana) zitholakala yonke indawo emazweni lapho ukutshala amazambane kwenziwa kuphela ngamapulazi amakhulu: i-USA, i-Netherlands, i-Denmark, njll. , kukhona futhi ukuhlukahluka okuphezulu kwe-genotypic ezingadini ezizimele. Ekupheleni kwekhulu lama-20, "imigqa yama-clonal" yayisakazeke ezindaweni zase-Asia naseMpumalanga Ekude yaseRussia (Elansky et al., 2001), okusobala ukuthi kungenxa yokusetshenziswa kwezinhlobo ezifanayo zokutshala amazambane awethu kuphela. ukukhiqiza. Muva nje, isimo kulezi zifunda siphinde saqala ukushintsha sibheke ekwandeni kokuhlukahluka kwe-genotypic yabantu.
Ukungabikho kokwelashwa okunamandla ngamalungiselelo esikhunta kunomunye umphumela oqondile - akukho ukuqoqwa kwezinhlobo ezimelana nezingadi. Ngempela, imiphumela yethu ikhombisa ukuthi izinhlobo ezimelana ne-metaxyl zitholakala kancane kakhulu ezingadini ezizimele kunasezitshalweni ezithengiswayo.
Ukusondela kokutshalwa kwezambane kanye notamatisi, okujwayelekile ezingadini ezizimele, kusiza ukufuduka kwezinhlobo phakathi kwalezi zitshalo, ngenxa yalokho, eminyakeni eyishumi edlule, phakathi kwezinhlobo ezihlukanisiwe namazambane, ingxenye yezinhlobo ezithwala isakhi sofuzo ukumelana. kuya ezinhlobonhlobo zikatamatisi we-cherry (T1), obekuyisici sangaphambilini "sohlobo" lukatamatisi kuphela. Izinhlobo ezinofuzo lwe-T1 ezimweni eziningi zinolaka kakhulu kukho kokubili amazambane notamatisi.
Eminyakeni yamuva, ukulimaza sekwephuzile kutamatisi kwaqala ukuvela ezimweni eziningi ngaphambili kunamazambane. Izithombo zikatamatisi zingangenwa ama-oospores enhlabathini, noma ama-oospores akhona embewu katamatisi noma anamathele kuwo (Rubin et al., 2001). Eminyakeni engu-15 edlule, inani elikhulu lembewu ehlanganisiwe eshibhile, ikakhulukazi ethunyelwa kwamanye amazwe, ibonakale ezitolo, okusetshenziswe iningi labakhiqizi abancane abashintshile. Imbewu ingaletha izinhlobo ezinama-genotypes ajwayelekile ezifundeni zokulinywa kwazo. Esikhathini esizayo, lezi genotypes zifakwe enqubweni yocansi emasimini angasese, okuholela ekuveleni kwama-genotypes amasha ngokuphelele.
Ngakho-ke, kungashiwo ukuthi izingadi zemifino ezizimele "ziyimbiza yokuncibilika" yomhlaba wonke, lapho, ngenxa yokushintshaniswa kwezinto zofuzo, ama-genotypes akhona acutshungulwa futhi avele amasha ngokuphelele. Ngaphezu kwalokho, ukukhethwa kwabo kwenzeka ezimweni ezihluke kakhulu kulezo ezidalelwe amazambane emapulazini amakhulu: ukungabikho komshini we-fungicidal, ukufana kwezinhlobonhlobo zokutshala, ukutholakala kwezitshalo ezithintekayo izinhlobo ezihlukahlukene zokutheleleka ngegciwane kanye namagciwane, ukusondelana notamatisi kanye ama-nightshades asendle, ukuwela okusebenzayo kanye nokwakheka kwe-oospore, ithuba lokuthi ama-oospores asebenze njengomthombo wokutheleleka ngonyaka olandelayo.
Konke lokhu kuholela ekuhlukeni okuphezulu kakhulu kwe-genotypic yabantu basemuva. Ezimweni ze-epiphytotics ezingadini zemifino, ukubola sekwephuzile kusakazeka ngokushesha okukhulu futhi kukhishwa inqwaba yezinhlamvu, indizela ezindaweni ezitshaliwe ezithengisayo. Kodwa-ke, njengoba ingene emkhakheni wezohwebo ngohlelo olufanele lobuchwepheshe bezolimo kanye nokuvikelwa kwamakhemikhali, izinhlayiya eziye zandiza azikwazi ukuqala i-epiphytotics ensimini, ngenxa yokungabikho kwemigqa ye-clonal ephikisana ne-fungicides ekhethekile. ezinhlobonhlobo ezitshaliwe.
Omunye umthombo we-inoculum eyinhloko kungase kube izilimo eziyizigaxa ezigulayo ezivaleleke ezitshalweni ezithengiswayo. Lezi nezilimo eziyizigaxa zazikhulile, njengomthetho, emikhakheni enobuchwepheshe obuhle bezolimo kanye nokuvikelwa kwamakhemikhali okunamandla. I-genotypes ye-isolates ethinte izilimo eziyizigaxa ijwayele ukuthuthukiswa kwezinhlobonhlobo zazo. Lezi zinhlobo ziyingozi kakhulu ekutshalweni kwezentengiselwano kune-inoculum evela ezingadini ezizimele. Lokhu kucabanga futhi kusekelwa imiphumela yocwaningo lwethu. Abantu abahlukanisiwe nezinkambu ezinkulu ezinokuvikelwa kwamakhemikhali okwenziwe kahle kanye nobuchwepheshe obuhle bezolimo abahlukani ngokuhlukahluka kwe-genotypic. Ngokuvamile lena imigqa yama-clonal amaningana anolaka kakhulu.
Izinhlobo zembewu ethengiswayo zingangena emiphakathini yasezivandeni zemifino futhi zibandakanyeke ezinhlelweni eziqhubekayo kuzo. Kodwa-ke, engadini yemifino, ukuncintisana kwabo kuyoba ngaphansi kakhulu kunensimu yezohwebo, futhi maduzane bazoyeka ukuba khona ngendlela yomugqa we-clonal, kodwa izakhi zabo zofuzo zingasetshenziswa kubantu "bengadi".
Ukutheleleka okuvela ezitshalweni "zokuzithandela" nasezinqumbini zezilimo eziyizigaxa eziqoqiwe ngesikhathi sokuvuna akufanelekile eRussia, ngoba Ezifundeni ezikhulayo zamazambane eRussia, ukuqandisa kwenhlabathi yasebusika okujulile kubonakala, futhi izitshalo ezivela kuma-tubers ezidlulele emhlabathini azivamile ukukhula. Ngaphezu kwalokho, njengoba ucwaningo lwethu lubonisa, i-pathogen ye-blight sekwephuzile ayiphili emazingeni okushisa amabi ngisho nasezigabeni ezigcine ukusebenza kwazo. Endaweni eyomile, lapho kutshalwa khona amazambane okuqala, ukubola kwakamuva kuyivelakancane ngenxa yenkathi eyomile neshisayo yokukhula.
Ngakho-ke, okwamanje sibheka ukuhlukaniswa kwezibalo ze-P. infestans zibe "insimu" kanye "nezingadi". Kodwa-ke, eminyakeni yamuva nje, kube nezinqubo eziholela ekuhlanganeni nasekungeneni kwezinhlobo ze-genotypes ezivela kulezi zizwe.
Phakathi kwabo, umuntu angaphawula ukwanda okujwayelekile kolwazi lokufunda nokubhala kwabakhiqizi abancane, ukuvela kwamaphakheji amancane amazambane athengekayo, ukusabalala kwamalungiselelo e-fungicidal emaphaketheni amancane, nokulahlekelwa kokwesaba "ikhemikhali" ngabantu.
Izimo ziphakama lapho, ngenxa yokusebenza ngamandla komhlinzeki oyedwa, amadolobhana wonke etshalwa izigaxa zembewu yohlobo olufanayo futhi anikezwe amaphakheji amancane ezibulala-zinambuzane ezifanayo. Kungacatshangwa ukuthi amazambane ezinhlobonhlobo ezifanayo azotholakala ezitshalweni ezithengiswayo eduze.
Ngakolunye uhlangothi, ezinye izinkampani ezihweba ngezibulala-zinambuzane zikhuthaza amasu okwelapha amakhemikhali “esabelomali”. Kulokhu, inani lemithi enconyiwe lithathwa kancane futhi kunikezwa ama-fungicides ashibhile, futhi ukugcizelelwa akukhona ukuvimbela ukuthuthukiswa kokulimala okuphuzile kuze kube ukugunda iziqongo, kodwa ekubambezelekeni okuthile kwe-epiphytoty ukuze kwandiswe isivuno. Izinhlelo ezinjalo zifanelekile ngokwezomnotho lapho kukhula amazambane e-ware kusuka kumbewu yezinga eliphansi, lapho empeleni kungekho mbuzo wokuthola isivuno esikhulu. Kodwa-ke, kulokhu, ngokungafani nenani labantu basengadini yemifino, isizinda sofuzo esilinganisiwe samazambane sinomthelela ekukhethweni kwezinhlanga ezithile zomzimba, eziyingozi kakhulu kulezi zinhlobonhlobo.
Ngokuvamile, ukuthambekela ekuhlanganeni kwezindlela "zengadi" kanye "nensimu" yokukhiqiza amazambane kubonakala kuyingozi kithi. Ukuvimbela imiphumela yabo emibi, endlini kanye nasemkhakheni wezohwebo, kuzodingeka kokubili ukulawula ukuhlukahluka kwamazambane embewu kanye nohlu lwama-fungicides olunikezwa abanikazi abazimele emaphaketheni amancane, nokulandelela izinhlelo zokuvikela amazambane kanye nokusetshenziswa. amalungiselelo okubulala isikhunta emkhakheni wezohwebo.
Ezindaweni zemboni yangasese, kukhona ukuthuthukiswa okujulile hhayi nje kuphela okulimazayo, kodwa ne-Alternaria. Iningi labanikazi bezindlu ezizimele abathathi izinyathelo ezikhethekile zokuvikela i-Alternaria, benza iphutha ngokuthuthukiswa kwe-Alternaria ngokubuna kwemvelo kwamahlamvu noma ukukhula kokubola sekwephuzile. Ngakho-ke, ngokuthuthuka okukhulu kwe-Alternaria ezinhlobonhlobo ezisengozini, iziza zasendlini zingasebenza njengomthombo we-inoculum wokutshalwa kwentengiso.
Izindlela zokuhlukahluka
Inqubo yokuguquguquka
Njengoba ukuvela kokuguqulwa kwezakhi zofuzo kuyinqubo engahleliwe eqhubeka nemvamisa ephansi, ukuvela koguquko kunoma iyiphi indawo kuncike ebuningini bokuguquguquka kwale ndawo kanye nobukhulu besibalo sabantu. Lapho kufundwa imvamisa yokuguqulwa kwezinhlobo ze-P. infestans, inani lamakholoni akhuliswa emithonjeni yezakhi ezikhethiwe ngemva kokwelashwa ngamakhemikhali noma okuguquguqukayo komzimba ngokuvamile kunqunywa. Njengoba kungabonwa kudatha ethulwe kuThebula 8, imvamisa yokuguqulwa kohlobo olufanayo ku-loci ehlukene ingahluka ngama-oda ambalwa obukhulu. Imvamisa ephezulu yokuguqulwa kokumelana ne-metataxyl ingaba esinye sezizathu zokunqwabelana kwezinhlobo ezimelana nayo ngokwemvelo.
Ukuvama kokuguquka kwezakhi zofuzo okuzenzakalelayo noma okubangelwayo, okubalwe ngesisekelo sokuhlolwa kwaselabhorethri, akuhambelani ngaso sonke isikhathi nezinqubo ezenzeka kubantu bemvelo, ngenxa yezizathu ezilandelayo:
1. Ekuqhekekeni kwe-nuclear asynchronous, akunakwenzeka ukulinganisa imvamisa yokuguqulwa kwesizukulwane ngasinye senuzi. Ngakho-ke, ukuhlola okuningi kunikeza ulwazi ngokuqondile kuphela mayelana nemvamisa yokuguqulwa kwezakhi zofuzo, ngaphandle kokuhlukanisa phakathi kwezenzakalo ezimbili eziguqukayo kanye nesenzakalo esisodwa esilandela i-mitosis.
2. Ukuguqulwa kwesinyathelo esisodwa kuvame ukunciphisa ibhalansi ye-genome, ngakho-ke, kanye nokutholwa kwendawo entsha, ukufaneleka okuvamile kwento ephilayo kuyehla. Iningi lezinguquko ezitholwe ngokuhlolwa zinolaka oluncishisiwe futhi azirekhodwa kubantu bemvelo. Ngakho-ke, i-coefficient yokuhlotshaniswa phakathi kwezinga lokumelana kwe-P. infestans mutants kuya ku-phenylamide fungicides kanye nezinga lokukhula endaweni yokwenziwa laliyisilinganiso (-0,62), kanye nokumelana nama-fungicides kanye nolaka emaqabunga amazambane (-0,65) (Derevyagina et al., 1993), okukhombisa ukufaneleka okuphansi kwezinguquko. Ukuguqulwa kokumelana ne-dimethomorph nakho kuhambisane nokwehla okubukhali kokusebenza (Bagirova et al., 2001).
3. Iningi lezinguquko ezizenzakalelayo nezibangelwayo ziyaphindaphindeka futhi azizivezi ngendlela evamile ekuhlolweni, kodwa zakha inqolobane efihlekile yokuhlukahluka kwabantu bemvelo. Izinhlobo eziguquguqukayo ezihlukanisiwe ekuhlolweni kwaselabhorethri zithwala uguquko olunamandla noma olunamandla (Kulish noDyakov, 1979). Ngokusobala, i-nuclear diploidy ichaza imizamo engaphumelelanga yokuthola izinto eziguquliwe ngaphansi kwethonya lemisebe ye-UV eyingozi ezinhlobonhlobo ezazikwazi ukumelana ngaphambili (McKee, 1969). Ngokwezibalo zombhali, ukuguqulwa okunjalo kungenzeka ngemvamisa engaphansi kwe-1: 500000. Ukuguqulwa kokuguqulwa kwezinguquko ku-homozygous, isimo esivezwe ngendlela emangalisayo kungenzeka ngenxa yokuhlangana kabusha kocansi noma okungafani nobulili (bona ngezansi). Kodwa-ke, nakulokhu, ukuguqulwa kungafihlwa yi-alleles evelele ye-nuclei yohlobo lwasendle ku-cenotic (multinucleated) mycelium futhi i-phenotypically fixed kuphela ngesikhathi sokwakhiwa kwama-zoospores e-mononuclear.
Ithebula 8. Ukuvama kwezinguquko ze-P. infestans kuya ezintweni ezivimbela ukukhula ngaphansi kwesenzo se-nitrosomethylurea (Dolgova, Dyakov, 1986; Bagirova et al., 2001)
Ukuxhumana | Imvamisa yokuguqula |
I-Oxytetracycline | 6,9 10 x-8 |
Blasticidin S | 7,2 x 10-8 |
I-Streptomycin | 8,3 x10-8 |
I-Trichothecin | 1,8 10 x-8 |
I-Cycloheximide | 2,1 10 x-8 |
I-Daaconil | <4 x 10-8 |
I-Dimethomorph | 6,3 10 x-7 |
I-Metalaxil | 6,9 10 x-6 |
Ubukhulu besibalo sabantu nabo badlala indima ebalulekile ekuveleni kokuguquka kwezakhi zofuzo okuzenzakalelayo. Kubantu abaningi kakhulu, lapho inani lamaseli N> 1 / a, lapho u-a kuyizinga lokuguqulwa kwezakhi zofuzo, ukuguqulwa kuyeka ukuba yinto engahleliwe (Kvitko, 1974).
Izibalo zibonisa ukuthi ngokuhlaselwa okumaphakathi kwensimu yamazambane (amabala angama-35 esitshalweni ngasinye), kwakheka izinhlamvu eziyi-8x1012 nsuku zonke ehektheleni elilodwa (i-Dyakov ne-Suprun, 1984). Ngokusobala, lezi zibalo ziqukethe zonke izinguquko ezivunyelwe uhlobo lokushintshana endaweni ngayinye. Ngisho noguquko olungajwayelekile, olwenzeka ngobuningi be-10-9, luzotholwa abantu abayinkulungwane ezigidini ezihlala ehektheleni elilodwa lensimu yamazambane. Ngoguquko olwenzeka ngokuvama okuphezulu (isibonelo, 10-6), esibalweni esinjalo, ukuguqulwa okuhlukahlukene okubhanqiwe kungenzeka nsuku zonke (ngasikhathi sinye kuma-loci amabili), i.e. inqubo yokuguqula izothatha indawo yokuhlanganisa kabusha.
Ukufuduka
Kuma-P. infestans, izinhlobo ezimbili eziyinhloko zokufuduka zaziwa: ukuvala amabanga (ngaphakathi kwensimu yamazambane noma amasimu angomakhelwane) ngokusabalalisa i-zoosporangia ngamaza omoya noma isifutho semvula, kanye namabanga amade - ngezilimo eziyizigaxa zokutshala noma izithelo zikatamatisi ezithuthwayo. Indlela yokuqala ihlinzeka ngokunwetshwa kokugxila kwesifo, okwesibili - ukudalwa kwe-foci entsha ezindaweni ezikude neyokuqala.
Ukusabalala kokutheleleka nge-tomato nezilimo eziyizigaxa nezithelo akugcini nje ukufaka isandla ekuveleni kwalesi sifo ezindaweni ezintsha, kodwa futhi kuwumthombo oyinhloko wokuhlukahluka kofuzo kubantu. Esifundeni saseMoscow, amazambane atshalwa, alethwa ezindaweni ezahlukahlukene zaseRussia naseNtshonalanga Yurophu. Izithelo zikatamatisi zilethwa ezindaweni eziseningizimu yeRussia (Isifunda sase-Astrakhan, i-Krasnodar Territory, eNyakatho yeCaucasus). Imbewu katamatisi, engasebenza njengemithombo yokutheleleka (Rubin et al., 2001), ibuye ingeniswe ezindaweni eziseningizimu yeRussia, China, amazwe aseYurophu nakwamanye amazwe.
Ngokwezibalo zika-E. Mayr (1974), izinguquko zofuzo kubantu bendawo okubangelwa ukuguquka kwezakhi zofuzo akuvamile ukuba zidlule i-10-5 endaweni ngayinye, kuyilapho ezindaweni ezivulekile, ukushintshaniswa ngenxa yokugeleza kwezakhi zofuzo okungenani ku-10-3 - 10-4.
Ukufuduka kwezilimo eziyizigaxa ezinegciwane kunomthelela ekungeneni kwama-P. infestans eYurophu, asakazeka kuzo zonke izifunda zomhlaba lapho kutshalwa khona amazambane; babangela izinguquko ezingathi sína kakhulu zabantu. Ukubola okuphuzile kumazambane kwavela endaweni yoMbuso WaseRussia cishe ngasikhathi sinye nokuvela kwawo eNtshonalanga Yurophu.
Njengoba lesi sifo saqala ukuphawulwa ngo-1846-1847 emazweni aseBaltic futhi eminyakeni eyalandela kuphela sasakazeka eBelarus nasezifundeni ezisenyakatho-ntshonalanga yeRussia, imvelaphi yaso yaseNtshonalanga Yurophu isobala. Umthombo wokuqala wesifo sekwephuzile eZweni Elidala awubonakali kangako. I-hypothesis eyakhiwe nguFry et al. (Fry et al., 1992; Fry, Goodwin, 1995, Goodwin et al., 1994) iphakamisa ukuthi i-parasite yaqala ukungena eNyakatho Melika isuka eMexico, lapho yasakazekela phezu kwezitshalo, yabe isithuthwa. ukuya eNtshonalanga Yurophu (umkhiwane 7).
Njengomphumela wokudonswa okuphindaphindiwe (umphumela ophindwe kabili we "bottleneck"), ama-clones angashadile afika eYurophu, inzalo yakho eyadala ubhubhane kuyo yonke indawo ye-Old World, lapho kutshalwa khona amazambane. Njengobufakazi balokhu kucatshangelwa, ababhali bacaphuna, okokuqala, ukuvela okukhona yonke indawo kohlobo olulodwa kuphela lokuhlangana (A1) futhi, okwesibili, ukufana kwe-genotypes yezinhlobo ezifundwayo ezivela ezifundeni ezahlukahlukene (zonke zisuselwa kumaka wamangqamuzana, okuhlanganisa 2 isozyme loci, DNA amaphethini zeminwe, kanye nokwakheka DNA mitochondrial kuyafana futhi kuhambisana clone US-1 echazwe e-USA). Kodwa-ke, enye idatha iphakamisa ukungabaza mayelana okungenani nezinye izihlinzeko ze-hypothesis eshiwo. Ukuhlaziywa kwe-P. infestans mitochondrial DNA ehlukanisiwe namasampula amazambane e-herbarium angenwe yileli gciwane ngesikhathi sokuqala se-epiphytotic ngawo-40s kwabonisa ukuthi ayahluka ngesakhiwo se-mitochondrial DNA kusuka ku-clone US-1, ngakho-ke, okungenani kwakungewona ukuphela komthombo wokutheleleka. eYurophu (Ristaino et al, 2001).
Isimo esinomonakalo ophuzile saba sibi futhi ngeminyaka yama-80s wekhulu lama-XX. Izinguquko ezilandelayo zenzeke:
I-1) Isilinganiso sobudlova sabantu siye sanda, okuye kwaholela, ikakhulukazi, ekusabalaleni okusabalele kwendlela eyingozi kakhulu yokulimaza sekwephuzile - ukulimala kwama-petioles neziqu.
2) Kwakukhona ushintsho ngesikhathi sokubonakala kokulimaza sekwephuzile kumazambane - kusukela ngasekupheleni kukaJulayi kuya ekuqaleni kukaJulayi ngisho nasekupheleni kukaJuni.
3) Uhlobo lokukhwelana lwe-A2, olwalungekho ngaphambilini eZweni Elidala, selutholakala yonke indawo.
Lezi zinguquko zandulelwa izehlakalo ezimbili: ukusetshenziswa okukhulu kwesikhunta esisha i-metaxyl (Schwinn kanye no-Staub, 1980) kanye nokuvela kweMexico njengomhwebi wamazambane emhlabeni wonke (Niederhauser, 1993). Ngokuvumelana nalokhu, kwabekwa izizathu ezimbili zokushintshwa kwenani labantu - ukuguqulwa kohlobo lokukhwelana ngaphansi kwethonya le-metaxyl (Ko, 1994) kanye nokwethulwa okukhulu kwezinhlobo ezintsha ezinama-tuber anegciwane avela eMexico (Fry and Goodwin, 1995). Nakuba ukuguqulwa kwezinhlobo zokukhwelana ngaphansi kwethonya le-metaxyl akutholwanga ngu-Ko kuphela, kodwa futhi emisebenzini eyenziwa elabhorethri yaseMoscow State University (Savenkova, Chherepennicova-Anikina, 2002), i-hypothesis yesibili ingcono. Kanye nokubonakala kohlobo lwesibili lokukhwelana, izinguquko ezinkulu zenzeka kuma-genotypes wezinhlobo zaseRussia ze-P. infestans, kuhlanganise nezakhi zofuzo ezingathathi hlangothi (i-isozyme ne-RFLP loci), kanye nesakhiwo se-mitochondrial DNA. Inkimbinkimbi yalezi zinguquko ayikwazi ukuchazwa ngesenzo se-metaxyl; kunalokho, kwaba nokungeniswa okukhulu kwezinhlobo ezintsha ezivela eMexico, okwathi, ngokuba nolaka (Kato et al., 1997), kwasusa izinhlobo ezindala (US-1) , iba namandla kubantu. Ushintsho ekubunjweni kwabantu baseYurophu lwenzeka ngesikhathi esifushane kakhulu - kusukela ngo-1980 kuya ku-1985 (Fry et al., 1992). Endaweni ye-USSR yangaphambili, "izinhlobo ezintsha" zatholakala emaqoqweni avela e-Estonia ngo-1985, okungukuthi, ngaphambi kwePoland naseJalimane (uGoodwin et al., 1994). Okokugcina "uhlobo oludala lwe-US-1" eRussia lwahlukaniswa notamatisi onegciwane esifundeni saseMoscow ngo-1993 (Dolgova et al., 1997). Futhi eFrance, izinhlobo “ezindala” zatholakala ekutshalweni kukatamatisi kuze kube sekuqaleni kwawo-90s, okungukuthi, ngemva kwesikhathi eside zanyamalala kumazambane (uLeberton no-Andrivon, 1998). Izinguquko ezinhlotsheni ze-P. infestans zithinte izici eziningi, okuhlanganisa nalezo ezibaluleke kakhulu ezingokoqobo, futhi zandisa ukulimaza kokulimaza sekwephuzile.
Ukuhlanganiswa kabusha kwezocansi
Ukuze ukuhlangana kabusha kocansi kube nomthelela ekuguquguqukeni, kuyadingeka, okokuqala, ukuba khona kwezinhlobo ezimbili zokuzalanisa kubantu ngesilinganiso esisondele ku-1: 1, futhi, okwesibili, ukuba khona kokuhlukahluka kwabantu bokuqala.
Isilinganiso sezinhlobo zokukhwelana siyehluka kakhulu kubantu abahlukene futhi ngisho naseminyakeni eyahlukene ngenani labantu abafanayo (Ithebula 9,10, 90). Izizathu zalolu shintsho olukhulu emazandleni ezinhlobo zokukhwelana kubantu (njengoba, ngokwesibonelo, eRussia noma kwa-Israyeli ekuqaleni kwawo-2002s ekhulwini leminyaka elidlule) azaziwa, kodwa kukholakala ukuthi lokhu kungenxa yokwethulwa kokuncintisana okwengeziwe. ama-clones (Cohen, XNUMX).
Enye idatha engaqondile ikhombisa inkambo yenqubo yocansi eminyakeni ethile nasezifundeni ezithile:
1) Ucwaningo lwezibalo zabantu abavela esifundeni saseMoscow lubonise ukuthi kubantu abangu-13 lapho isabelo sohlobo lokukhwelana i-A2 sasingaphansi kuka-10%, ingqikithi yokuhlukahluka kofuzo ebalwe kuma-isozyme loci amathathu kwaba ngu-0,08, futhi kubantu abangu-14 lapho isabelo sika-A2 kudlule ama-30%, ukuhlukahluka kwezakhi zofuzo kwakuphindwe kabili (0,15) (Elansky et al., 1999). Ngakho, lapho ephakama amathuba okuba nobuhlobo bobulili, ayanda ukuhlukahluka kwezakhi zofuzo kubantu.
2) Ubudlelwano phakathi kwesilinganiso sezinhlobo zokukhwelana kubantu kanye nokuqina kokwakheka kwe-oospore babonwa kwa-Israel (Cohen et al., 1997) naseHolland.
(Flier et al., 2004). Ucwaningo lwethu lukhombisile ukuthi kubantu lapho ukuhlukaniswa nohlobo lokuhlangana kwe-A2 kubalwa ku-62, 17, 9, no-6%, ama-oospores atholakala ku-78, 50, 30, kanye no-15% wamaqabunga amazambane ahlaziywe (anama-2 noma amabala amaningi), ngokulandelana.
Amasampula anamachashazi angu-2 noma ngaphezulu ngokuvamile aqukethe ama-oospores kunamasampuli anendawo engu-1 (32 kanye no-14% wamasampuli, ngokulandelana) (Apryshko et al., 2004).
Ama-Oospores ayevame kakhulu emaqabunga engqimba ephakathi nendawo ephansi yesitshalo samazambane (Mytsa et al., 2015; Elansky et al., 2016).
3) Kwezinye izifunda, kutholwe ama-genotypes ahlukile, okwenzeka okuhlotshaniswa nokuhlangana kabusha kocansi. Ngakho-ke, ePoland ngo-1989 naseFrance ngo-1990, i-homozygous ihlunga i-glucose-6-.
i-phosphate isomerase (GPI 90/90). Njengoba ngaphambilini kwakuhlangatshezwana nama-heterozygote angama-10/90 iminyaka eyi-100, i-homozygosity kuthiwa ibangelwa ukuhlangana kabusha kocansi (Sujkowski et al., 1994). E-Colombia (USA), ukuhlukaniswa okuhlanganisa i-A2 ne-GPI 100/110 kanye ne-A1 ne-GPI 100/100 kuvamile, nokho, ekupheleni kwesizini ka-1994 (Agasti 16 noSepthemba 9), izinhlobo ezine-recombinant genotypes (A1 GPI 100/ 110 kanye ne-A2 GPI 100/100) (Miller et al., 1997).
4) Kwezinye izakhamuzi zasePoland (Sujkowski et al., 1994) kanye naseNyakatho yeCaucasus (Amatkhanova et al., 2004), ukusatshalaliswa kwe-DNA loci yeminwe kanye ne-allozyme protein loci kuhambisana nokusatshalaliswa kwe-Hardy-Weinberg, okubonisa
mayelana nesabelo esiphezulu somnikelo wokuhlanganisa ucansi kabusha ekushintshashintsheni kwabantu. Kwezinye izifunda zaseRussia, akukho ukuxhumana okusatshalaliswa kwe-Hardy-Weinberg kubantu okwatholakala, kodwa ukuba khona kokungalingani kokuxhumana kwaboniswa, okubonisa ukuvela kokuzala kwama-clonal (Elansky et al., 1999).
5) Ukuhlukahluka kofuzo (GST) phakathi kwezinhlobo ezinezinhlobo ezihlukene zokukhwelana (A1 kanye ne-A2) bekuphansi kunaphakathi kwabantu abahlukene (Sujkowski et al., 1994), okubonisa ngokungaqondile iziphambano zocansi.
Ngesikhathi esifanayo, umnikelo wokuhlanganisa ubulili kabusha ezinhlobonhlobo zabantu awukwazi ukuba phezulu kakhulu. Lo mnikelo ubalelwe abantu besifunda saseMoscow (Elansky et al., 1999). Ngokwezibalo zika-Lewontin (1979) "ukuhlanganisa kabusha, okungaveza okuhlukile okusha kusuka ku-loci ezimbili ne-frequency engeqi umkhiqizo we-heterozygosities yabo, kusebenza kuphela uma amanani we-heterozygosity kuwo womabili ama-alleles asevele ephezulu."
Ngesilinganiso sezinhlobo ezimbili zokumatanisa, okujwayelekile esifundeni saseMoscow, esilingana no-4: 1, imvamisa yokuhlanganisa izoba ngu-0,25. Amathuba okuthi okunqamule izinhlobo kuzoba yi-heterozygous kokubili kokuthathu okuhloliwe isozygous loci kubantu abahloliwe kwaba ngu-0,01 (izinhlobo ezingu-2 kwezingu-177). Ngakho-ke, amathuba okuba khona kwama-heterozygote aphindwe kabili ngenxa yokuhlanganiswa kabusha akufanele adlule umkhiqizo wawo aphindwe ngamathuba okuwela (0,25x0,02x0,02) = 10-4, i.e. ama-recombinants ocansi ngokuvamile awaweli kusampula elihloliwe lezinhlobo. Lezi zibalo zenzelwe abantu abavela esifundeni saseMoscow, ezibonakala ngokuhlukahluka okuphezulu. Ezixukwini ze-monomorphic ezifana nezaseSiberia, inqubo yocansi, ngisho noma ivela kubantu ngabanye, ayikwazi ukuthonya ukuhlukahluka kwabo kofuzo.
Ukwengeza, i-P. infestans ibonakala ngokungahambi kahle kwe-chromosome ku-meiosis, okuholela ku-aneuploidy (Carter et al., 1999). Izifo ezinjalo zinciphisa ukuvunda kwama-hybrids.
I-Parasexual recombination, ukuguqulwa kofuzo lwe-mitotic
Ekuhlolweni kokuhlukaniswa kwezinhlobo ze-P. infestans ezinokuguqulwa kokumelana nama-inhibitors ahlukene okukhula, kwatholakala ukuvela kwama-misolates amelana nawo womabili ama-inhibitors (Shattock and Shaw, 1975; Dyakov, Kuzovnikova, 1974; Kulish, Dyakov,
1979). Izifo ezimelana nama-inhibitors amabili okukhula zavela ngenxa ye-heterokaryotization ye-mycelium, futhi kulokhu, zaqhekeka ngesikhathi sokuzalanisa ama-zoospores e-mononuclear (Judelson, Ge Yang, 1998), noma awazange ahlukane enzalweni ye-monozoosporous, ngoba yayine-tetraploid ( njengoba ama-isolate okuqala angama-diploid) ama-nuclei (K, 1979). I-Heterozygous diploid ihlukaniswe ngemvamisa ephansi kakhulu ngenxa ye-haploidization, i-chromosome nondisjunction, kanye nokuwela imitotic (Poedinok et al., 1982). Ukuvama kwalezi zinqubo kungandiswa ngosizo lwemiphumela ethile kuma-heterozygous diploid (isibonelo, imisebe ye-UV yezinhlamvu ezikhulayo).
Yize ukwakheka kwama-hybridi emifino anokumelana okuphindwe kabili akwenzeki kuphela ku-vitro, kodwa nasezigabeni zamazambane ezitheleleke ngengxube ye-mutants (Kulish et al., 1978), kunzima kakhulu ukuhlola indima ye-parasexual recombination esizukulwaneni se-parasexual recombination. ama-genotypes amasha kubantu. Imvamisa yokwakheka kwama-segregant ngenxa ye-haploidization, ukungahlangani kwama-chromosome kanye nokuwela i-mitotic ngaphandle kwemiphumela ekhethekile ayinakwa (ngaphansi kuka-10-3).
Ukwenzeka kwama-homozygous segregants ezinhlobo ze-heterozygous kungase kusekelwe kukho kokubili ukuwela i-mitotic cross over kanye nokuguqulwa kofuzo lwe-mitotic, okuthi ku-P. sojae kwenzeka ngokuvama kuka-3 x 10-2 kuya ku-5 x 10-5 endaweni ngayinye, kuye ngobunzima ( Chamnanpunt et al. , 2001).
Nakuba imvamisa yokwenzeka kwama-heterokaryons kanye ne-heterozygous diploid ivele yaba phezulu ngokungalindelekile (ifinyelele amaphesenti angamashumi), le nqubo yenzeka kuphela lapho amasiko aguqukayo atholwe ohlotsheni olufanayo ehlukaniswa. Uma usebenzisa izinhlobo ezahlukene ezihlukanisiwe nemvelo, i-heterokaryotization ayenzeki (noma yenzeka ngemvamisa ephansi kakhulu) ngenxa yokuba khona kokungahambisani kwezitshalo (Poedinok noDyakov, 1981; Anikina et al., 1997b; Cherepennikova-Anikina et al., 2002) ). Ngakho-ke, indima yokuhlanganiswa kabusha kwe-parasexual ingancishiswa kuphela ekuhlanganisweni kwe-intraclonal ku-nuclei ye-heterozygous kanye nokuguqulwa kofuzo olulodwa kuya esimweni se-homozygous ngaphandle kwenqubo yocansi. Le nqubo ingase ibe ngokubaluleka kwe-epidemiological ezinhlotsheni ezinokuguqulwa okuphindaphindekayo noma okunamandla okuphikiswa kwesikhunta. Ukuguqulwa kwayo esimweni se-homozygous ngenxa yenqubo ye-parasexual kuzokwandisa ukumelana komthwali we-mutation (Dolgova, Dyakov, 1986).
Ukwethulwa kofuzo
Izinhlobo ze-Heterothallic Phytophthora ziyakwazi ukuzalanisa ngokwakhiwa kwama-oospores ayingxube (bona i-Vorob'eva kanye neGridnev, 1983; Sansome et al., 1991; Veld et al., 1998). Ingxube yemvelo yezinhlobo ezimbili ze-Phytophthora yayinolaka kangangokuthi yabulala izinkulungwane zama-alder e-UK (Brasier et al., 1999). I-P. infestans ingenzeka nezinye izinhlobo ze-genus (P. erythroseptica, P. nicotianae, P. Cactorum, njll.) ezitshalweni ezivamile ezibamba umhlabathi kanye nomhlabathi, kodwa kunolwazi oluncane ezincwadini mayelana nokuba khona kwe-interspecific hybrids. . Ezimweni zaselabhorethri, ama-hybrids atholakala phakathi kwe-P. infestans kanye ne-P. Mirabilis (Goodwin and Fry, 1994).
Ithebula 9. Ingxenye yezinhlobo ze-P. infestans ezinohlobo lokukhwelana lwe-A2 emazweni ahlukene omhlaba esikhathini esisuka ku-1990 kuya ku-2000 (ngokolwazi lwemithombo yezincwadi ezivulekile nezingosi www.euroblight.net, www.eucablight.org)
izwe | 1990 | 1991 | 1992 | 1993 | 1994 | 1995 | 1996 | 1997 | 1998 | 1999 | 2000 |
---|---|---|---|---|---|---|---|---|---|---|---|
Belarus | 33 (12) | 34 (29) | |||||||||
Belgium | 15 (49 *) | 6 (66) | 20 (86) | ||||||||
Ecuador | 0 (13) | 0 (12) | 0 (19) | 0 (21) | 12 (41) | 25 (39) | 15 (75) | 22 (73) | 25 (68) | 0 (35) | |
Estonia | 8 (12) | ||||||||||
E-England | 4 (26) | 3 (630) | 9 (336) | ||||||||
Finland | 0 (15) | 19 (117) | 12 (16) | 21 (447) | 6 (509) | 9 (432) | 43 (550) | ||||
France | 0 (35) | 0 (56) | 0 (83) | 0 (67) | 0 (86) | 2 (135) | 7 (156) | 6 (123) | 0 (73) | 0 (285) | 0 (135) |
Hungary | 72 (32) | ||||||||||
Ирландия | 4 (145) | ||||||||||
Enyakatho. I-Ireland | 10 (41) | 9 (58) | 1 (106) | 0 (185) | 0 (18) | 0 (56) | 0 (35) | 0 (26) | |||
Netherlands | 7 (41) | 5 (276) | 24 (377) | 44 (353) | 23 (185) | ||||||
Norway | 25 (446) | 28 (156) | 8 (39) | 18 (257) | 38 (197) | ||||||
Peru | 0 (34, 1984 -86) | 0 (287, 1997-98) | 0 (112) | 0 (66) | |||||||
Poland | 19 (180) | 21 (142) | 33 (256) | 26 (149) | 35 (70) | ||||||
EScotland | 25 (147) | 11 (163) | 22 (189) | 5 (22) | |||||||
Sweden | 25 (263) | 62 (258) | 49 (163) | ||||||||
I-Wales | 0 (16) | 7 (97) | 0 (48) | 0 (25) | |||||||
Korea | 36 (42) | 10 (130) | 15 (98) | ||||||||
China | 20 (142, 1995-98) | 0 (6) | 0 (8) | 0 (35) | |||||||
Колумбия | 0 (40, 1994-2000) | ||||||||||
I-Uruguay | 100 (25, 1998-99) | ||||||||||
Марокко | 60 (108, 1997-2000) | 52 (25) | 42 (40) | ||||||||
Serbia | 76 (37) | ||||||||||
Мексика (Toluca) | 28 (292, 1988-89) | 50 (389, 1997-98) |
Ithebula 10. Ingxenye yezinhlobo ze-P. infestans ezinohlobo lokukhwelana lwe-A2 emazweni ahlukene emhlabeni kusukela ngo-2000 kuya ku-2011.
izwe | 2001 | 2002 | 2003 | 2004 | 2005 | 2006 | 2007 | 2008 | 2009 | 2010 | 2011 |
---|---|---|---|---|---|---|---|---|---|---|---|
Austria | 65 (83) | ||||||||||
Belarus | 42 (78) | ||||||||||
Belgium | 20 (102 *) | 4 (32) | 50 (14) | 25 (16) | 62 (13) | 54 (26) | 70 (54) | 30 (23) | 29 (35) | 62 (71) | 45 (49) |
Switzerland | 89 (19) | ||||||||||
Czech Republic | 35 (31) | 54 (64) | 38 (174) | 12 (80) | |||||||
Germany | 95 (53) | ||||||||||
Denmark | 48 (52) | ||||||||||
Ecuador | 5 (178) | 6 (108) | 9 (121) | 18 (94) | 2 (44) | 0 (66) | 5 (47) | ||||
Estonia | 54 (25) | 0 (24) | 33 (62) | 45 (140) | 25 (100) | 12 (103) | |||||
E-England | 4 (47) | 10 (96) | 31 (55) | 55 (790) | 68 (862) | 70 (552) | 68 (299) | ||||
Finland | 47 (162) | 12 (218) | 42 | ||||||||
France | 0 (186) | 4 (108) | 8 (61) | 22 (103) | 33 (303) | 65 (378) | 74 (331) | 75 (125) | 75 (12) | ||
Hungary | 48 (27) | 48 (90) | 9 | 7 | |||||||
Enyakatho. I-Ireland | 0 (38) | 0 (58) | 0 (40) | 0 (24) | 5 (54) | 0 (18) | 27 (578) | 45 (239) | 36 (213) | 82 (60) | 10 (80) |
Netherlands | 66 (24) | 93 (15) | 91 (11) | ||||||||
Norway | 39 (328) | 3 (115) | 12 (19) | ||||||||
Peru | 0 (36) | ||||||||||
Poland | 25 (46) | 10 (30) | 85 (20) | 38 (44) | 75 (66) | 55 (56) | 65 (35) | 72 (81) | 85 (21) | ||
EScotland | 3 (213) | 2 (474) | 24 (135) | 86 (337) | 88 (386) | 74 (172) | |||||
Sweden | 60 (277) | 39 (87) | |||||||||
Slovakia | 0 (36) | 14 (26) | 62 (26) | 0 (26) | |||||||
I-Wales | 25 (12) | 68 (106) | 80 (88) | 92 (143) | 75 (45) | ||||||
Korea | 46 (26) | ||||||||||
Brazil | 0 (49) | 0 (30) | |||||||||
China | 10 (30) | 0 (6) | 0 (6) | ||||||||
Вьетнам | 0 (294, 2003-04) | ||||||||||
E-Uganda | 0 (8) |
Amandla okwakheka kwe-genotypic yabantu
Izinguquko ekwakhekeni kwe-genotypic yezibalo ze-P. infestans zingenzeka ngaphansi kwethonya lokufuduka kwama-clones amasha esuka kwezinye izifunda, izinqubo zezolimo (ukushintsha kwezinhlobo, ukusetshenziswa kwemithi yokukhunta) kanye nezimo zezulu. Amathonya angaphandle athinta ama-clone ahlukene ezigabeni ezihlukene zomjikelezo wempilo; ngakho-ke, isibalo sabantu minyaka yonke siba nezinguquko zomjikelezo kumaza ofuzo angaphansi kokukhethwa, ngenxa yoshintsho endimeni eyinhloko yokukhukhuleka nokukhetha kwezakhi zofuzo.
Ithonya lezinhlobonhlobo
Ama-cultivar amasha anezakhi zofuzo ezisebenzayo zokuphikiswa okuqondile (R-genes) iyisici esinamandla esikhethayo esikhetha ama-clones anezakhi zofuzo ezihambisanayo ze-virulence kubantu be-P. infestans. Uma kungekho ukumelana okungaqondile ezinhlobonhlobo zamazambane, okuvimbela ukukhula kwenani le-pathogen, inqubo yokushintsha ama-clones avelele kubantu yenzeka ngokushesha okukhulu. Ngakho-ke, ngemva kokusabalala esifundeni saseMoscow sezinhlobonhlobo ze-Domodedovsky, ezinofuzo lokumelana no-R3, imvamisa yama-clones ayingozi yalolu hlobo lenyuke lisuka ku-0,2 laya ku-0,82 ngonyaka owodwa (Dyakov, Derevjagina, 2000).
Kodwa-ke, ukuguqulwa kwamaza wezakhi zofuzo ze-virulence (pathotypes) kubantu akwenzeki kuphela ngaphansi kwethonya lezinhlobo zamazambane ezitshaliwe. Isibonelo, eBelarus kuze kube ngu-1977, ama-clones ane-virulence gene 1 no-4 ayebusa, okubangelwa ukutshalwa kwezinhlobonhlobo zamazambane ezinofuzo lokumelana no-R1 no-R4 (Dorozhkin, Belskaya, 1979). Kodwa-ke, ekupheleni kweminyaka yama-70s wekhulu lama-XX, ama-clones avela anezinhlobo ezahlukene ze-virulence nezinhlanganisela zawo, futhi izakhi zofuzo ezihambisanayo azizange zisetshenziswe ekuzaleni amazambane (izakhi zofuzo ze-virulence eyengeziwe) (Ivanyuk et al., 2002). Isizathu sokuvela kwama-clones anjalo, ngokusobala, kungenxa yokufudukela eYurophu kwezinto ezithathelwanayo ezivela eMexico nezilimo eziyizigaxa zamazambane. Ekhaya, lawa ma-clones awazange athuthuke kumazambane atshaliwe kuphela, kodwa nasezinhlotsheni zasendle ezithwala izinhlobo ezihlukahlukene zofuzo; ngakho-ke, ukuhlanganiswa kwezakhi zofuzo eziningi ze-virulence ku-genome kwakudingeka ukuze kuphile kulezo zimo.
Ngokuqondene nezinhlobonhlobo ezinokumelana okungaqondile, bona, ngokunciphisa izinga lokuzala kabusha kwe-pathogen, babambezela ukuvela kwabantu bayo, okuyinto, njengoba sekushiwo kakade, kuwumsebenzi wenombolo. Njengoba ulaka luyi-polygenic, ama-clones aqukethe inani elikhulu lezakhi zofuzo "zobudlova" anqwabelana ngokushesha, lapho inani labantu liphakama. Ngakho-ke, izinhlanga ezinolaka kakhulu akuwona umkhiqizo wokujwayela izinhlobo ezitshaliwe ezinokumelana okungaqondile, kodwa, ngokuphambene nalokho, zivame ukutholwa ekutshalweni kwezinhlobo ezisengozini kakhulu ezingaba-ccumulators bezinhlamvu ze-parasite.
Ngakho, eRussia, abantu abanolaka kakhulu be-P. Infestans batholakala ezindaweni zama-epiphytoties wonyaka (abantu abavela ezindaweni zaseSakhalin, eLeningrad, naseBryansk). Ulaka lwalaba bantu luvele lwaba phezulu kunabaseMexico (Filippov et al., 2004).
Ngaphezu kwalokho, ama-oospores ambalwa akhiwa emaqabunga ezinhlobonhlobo ezimelana nalawo athintekayo (Hanson and Shattock, 1998), okusho ukuthi, ukumelana okungaqondile kwezinhlobonhlobo nakho kunciphisa ikhono lokuhlanganisa kabusha i-parasite kanye nethuba lezinye izindlela zasebusika.
Ithonya lama-fungicides
Ama-fungicides awagcini nje ukunciphisa inani lesikhunta se-phytopathogenic, i.e. kuthinte izici zobuningi bezibalo zabo, kodwa futhi kungashintsha ama-frequencies we-genotypes ngayinye, i.e. nomthelela ekubunjweni kwekhwalithi yabantu. Phakathi kwezinkomba ezibaluleke kakhulu zezibalo zabantu ezishintshayo ngaphansi kwethonya lama-fungicides yilezi ezilandelayo: izinguquko zokumelana nama-fungicides, izinguquko ezinolaka kanye ne-virulence, nezinguquko ezinhlelweni zokuzala.
Ithonya lama-fungicides ekuphikiseni kanye nolaka lwabantu
Izinga lalo mphumela linqunywa, okokuqala, ngohlobo lwesikhunta esisetshenzisiwe, esingahlukaniswa ngokwemibandela sibe yi-polysite, i-oligosite ne-monosite.
Iqembu lokuqala lihlanganisa ama-fungicides amaningi okuxhumana. Ukumelana nazo (uma kungenzeka nhlobo) kulawulwa inani elikhulu lezakhi zofuzo ezivezwayo ezibuthakathaka kakhulu. Lezi zakhiwo zinquma ukungabi bikho kwezinguquko ezibonakalayo ekuphikisweni kwenani labantu ngemva kokwelashwa kwayo ngama-fungicides (nakuba kwezinye izivivinyo kutholwe ukwanda kokungazweli). Isibalo sesikhunta esilondolozwe ngemuva kokufafazwa ngama-fungicides othintana nawo siqukethe amaqembu amabili ezinhlobo:
1) Izifo ezigcinwe ezindaweni zezitshalo ezingaphathwanga ngomuthi. Njengoba kwakungekho ukuthintana nesibulali sikhunta, ulaka nokumelana kwalezi zinhlobo akushintshi.
I-2) Izinkinga ezithintana ne-fungicide, ukugxila kwayo ezindaweni zokuxhumana kwakungaphansi kunokubulala. Njengoba kushiwo ngenhla, ukumelana kwale ngxenye yabantu nakho akushintshi, noma kunjalo, ngenxa yomphumela olimazayo we-fungicide ngisho nasekugxilweni okungaphansi kwe-metabolism ye-fungal cell, ukuqina okujwayelekile kanye nengxenye yayo ye-parasitic, ulaka, ukunciphisa (Derevyagina noDyakov, 1990).
Ngakho-ke, ngisho nengxenye yabantu abangakafi, evezwe ekuthinteni isikhunta, inolaka olubuthakathaka futhi ayikwazi ukuba umthombo we-epiphytotics. Ngakho-ke, ukucubungula ngokucophelela, okunciphisa imvamisa yengxenye yabantu abangaxhumani ne-fungicide, kuyisimo sokuphumelela kwezinyathelo zokuzivikela. Ukumelana ne-oligosite fungicides kulawulwa yizakhi zofuzo ezingeziwe.
Ukuguqulwa kwesakhi sofuzo ngasinye kuholela ekwenyukeni okuthile kokumelana, futhi izinga eliphelele lokumelana libangelwa ukwengezwa kokuguqulwa okunjalo. Ngakho-ke, ukwanda kokumelana kwenzeka kancane kancane. Isibonelo sokwenyuka kancane kancane kokumelana nokuguquka kokumelana ne-fungicide dimethomorph, esetshenziswa kakhulu ukuvikela amazambane ekubhubhiseni sekwephuzile. Ukumelana ne-Dimethomorph kuyi-polygenic kanye nesengezo. Ukuguqulwa kwesinyathelo esisodwa kukhulisa kancane ukumelana.
Ukuguqulwa ngakunye okulandelanayo kunciphisa usayizi oqondiwe futhi, ngenxa yalokho, imvamisa yezinguquko ezilandelayo (Bagirova et al., 2001). Ukwanda kokungazweli okumaphakathi kwesibalo sabantu ngemuva kokwelashwa okuningi nge-oligosite fungicide kwenzeka ngokwesinyathelo futhi kancane kancane. Izinga lale nqubo linqunywa okungenani izici ezintathu: imvamisa yokuguquguquka kwezakhi zofuzo zokumelana, i-coefficient of resistance (isilinganiso somthamo obulalayo wohlobo olunamandla olumelana noluzwelayo), kanye nomthelela wokuguqulwa kwezakhi zofuzo. izakhi zofuzo zokuqina.
Imvamisa yokwenzeka kokuguqulwa ngakunye okulandelayo iphansi kunangaphambili; ngakho-ke, inqubo inohlamvu oludambisayo (Bagirova et al., 2001). Kodwa-ke, uma izinqubo zokuhlanganisa kabusha (zocansi noma i-parasexual) zenzeka kubantu, khona-ke kungenzeka ukuhlanganisa ukuguqulwa okuhlukile kwabazali ohlotsheni oluhlanganisiwe futhi kusheshiswe inqubo. Ngakho-ke, abantu be-panmix bathola ukumelana ngokushesha kune-agamic, futhi ekugcineni, inani labantu abangenazo izithiyo zokungahambisani nezitshalo ngokushesha kunabantu abahlukaniswe yizithiyo ezinjalo. Mayelana nalokhu, ukuba khona kwezinhlobo zabantu ezihlukene ngezinhlobo zokukhwelana kusheshisa inqubo yokuthola ukumelana nama-oligosite fungicides.
Izinto zesibili nezesithathu azifaki isandla ekuqongeleleni okusheshayo kwezinhlobo ezimelana ne-dimethomorph kubantu. Ukuguqulwa ngakunye okulandelayo cishe kuphinda kabili ukumelana, okungasho lutho, futhi ngesikhathi esifanayo kunciphisa kokubili izinga lokukhula endaweni yokwenziwa kanye nolaka (Bagirova et al., 2001; Stem, Kirk, 2004). Mhlawumbe, ngakho-ke, phakathi kwezinhlobo zemvelo ze-P. infestans, ngisho nalezo eziqoqwe ezitshalweni zamazambane eziphathwe nge-dimethomorph, cishe azikho izinhlobo ezimelana nazo.
Isibalo sabantu abalashwa nge-oligosite fungicide sizophinde sibe namaqembu amabili ezinhlobo: lezo ezingakaze zihlangane ne-fungicide, ngakho-ke azizange zishintshe izici zokuqala (uma izinhlobo ezimelana zitholakala kuleli qembu, ngeke ziqongelele ekuqineni okuphezulu kanye nokuncintisana kwezinhlobo ezizwelayo), kanye nezinkinga ezithintana nokugxilisa okuyingozi kwesibulala-sikhunta. Kuphakathi kokugcina ukuthi ukuqoqwa kwezinhlobo eziphikisanayo kungenzeka, ngoba lapha zinezinzuzo ngaphezu kwalezo ezibucayi.
Ngakho-ke, lapho usebenzisa ama-oligosite fungicides, akukona ukucubungula ngokucophelela okubalulekile njengokugxila okuphezulu komuthi, izikhathi ezimbalwa eziphakeme kunomthamo obulalayo, ngoba nge-stepwise mutagenesis, ukumelana kokuqala kwezinhlobo eziguquliwe kuphansi.
Okokugcina, ukuguqulwa kokumelana ne-monosite fungicides kucacile kakhulu, okungukuthi, ukuguqulwa okukodwa kungabika izinga eliphezulu lokumelana kuze kufike ekulahlekelweni okuphelele kokuzwela. Ngakho-ke, ukwanda kokumelana kwabantu kwenzeka ngokushesha okukhulu.
Isibonelo semithi enjalo yokubulala isikhunta i-phenylamide, kuhlanganise nesikhunta esivame kakhulu, i-metaxyl. Ukuguqulwa kokumelana nakho kwenzeka ngemvamisa ephezulu, futhi izinga lokumelana nezinguquko liphezulu kakhulu - lidlula uhlobo olubucayi ngesilinganiso senkulungwane noma ngaphezulu (Derevyagina et al., 1993). Yize izinga lokukhula kanye nolaka lweziguquli eziguqukayo ezimelana nazo kuncipha ngokumelene nesizinda sokufa kwezinhlobo ezithintekayo ezivela ku-systemic fungicide, inani labantu eliphikisayo likhula ngokushesha futhi, ngokuhambisanayo, ulaka lwalo luyakhula. Ngakho-ke, ngemva kweminyaka eminingana yokusebenzisa i-fungicide, ulaka lwezinhlobo ezimelana nazo azikwazi nje ukulingana nolaka lwalezo ezibucayi, kodwa futhi zidlule (Derevyagina noDyakov, 1992).
Imithelela ekuhlanganiseni kabusha kocansi
Njengoba ukuvela kaningi kohlobo lokukhwelana lwe-A2 kubantu be-P. infestans kwaqondana nokusetshenziswa kakhulu kwe-metaxyl ngokumelene nokulimaza sekwephuzile, kwacatshangwa ukuthi i-metataxyl idala ukuguqulwa kohlobo lokukhwelana. Ku-P. parasitica, ukuguqulwa okunjalo ngaphansi kwesenzo se-Chloroneb ne-metatalaxyl kwafakazelwa ngokuhlolwa (Ko, 1994). Iphaseji eyodwa endaweni ene-metataxyl ephansi iholele ekuveleni kwe-homothallic isolate kusuka ohlotsheni lwe-P. infestans ezwelayo ku-metataxyl enohlobo lokukhwelana u-A1 (Savenkova kanye ne-Cherepnikova-Anikina, 2002). Ezindaweni ezilandelanayo kumidiya ene-metataxyl ephakeme kakhulu, akukho nesisodwa esisodwa esinohlobo lokumatanisa lwe-A2 esatholakala; nokho, ama-isolate amaningi, lapho ewela ngama-A2 isolate, esikhundleni sama-oospores, akhe ama-mycelium amabi futhi awanyumba. Iziqephu zohlobo olumelana nohlobo lokuhlangana kwe-A2 kumidiya ene-metataxyl ephezulu zisivumele ukuthi sithole izinhlobo ezintathu zezinguquko zohlobo lokukhwelana: 1) ubunyumba obuphelele lapho siwela nge-A1 kanye ne-A2 yodwa; 2) i-homotallism (ukwakheka kwe-oospores ku-monoculture); 3) ukuguqulwa kohlobo lokukhwelana lwe-A2 lube ngu-A1. Ngakho-ke, i-metaxyl ingabangela izinguquko ezinhlotsheni zokukhwelana emiphakathini ye-P. infestans futhi, ngenxa yalokho, ukuvela kokuhlangana kabusha kocansi kuzo.
Ithonya ekuhlanganiseni kabusha kwezitshalo
Amanye amajini okumelana nama-antibiotic andise imvamisa ye-hyphal heterokaryotization kanye ne-nuclear diploidization (Poedinok noDyakov, 1981). Njengoba kuphawuliwe ngaphambili, i-heterokaryotization ye-hyphae ngesikhathi sokuhlanganiswa kwezinhlobo ezahlukene ze-P. infestans zenzeka kancane kakhulu ngenxa yesimo sokungahambisani kwemifino kule fungus. Kodwa-ke, izakhi zofuzo zokumelana namanye ama-antibiotics zingaba nemiphumela engemihle, evezwa ekunqobeni ukungahambisani kwezitshalo. Lesi sakhiwo sasiphethwe yi-1S-1 mutant streptomycin resistance gene. Ukuba khona kwalezi zimo eziguquguqukayo emkhakheni we-phytophthora kungakhuphula ukugeleza kwezakhi zofuzo phakathi kwezinhlobo futhi kusheshise ukujwayelana nenani labantu ezinhlobonhlobo ezintsha noma ama-fungicides.
Amanye ama-fungicides kanye nama-antibiotics angathinta imvamisa ye-mitotic recombination, engaphinda iguqule amaza ohlobo lwe-genotype kubantu. I-benomyl esetshenziswa kabanzi yokubulala isikhunta ibophezela ku-beta-tubulin, iphrotheni okwakhiwa kuyo ama-microtubules e-cytoskeleton, futhi ngaleyo ndlela iphazamise izinqubo zokuhlukaniswa kwe-chromosome ku-anaphase ye-mitosis, okwandisa imvamisa ye-mitotic recombination (Hastie, 1970).
I-fungicide para-fluorophenylalanine, esetshenziselwa ukwelapha isifo sase-Dutch kuma-elms, inendawo efanayo. I-Para-fluorophenylalanine yandisa imvamisa yokuhlanganiswa kabusha ku-heterozygous diploids P. infestans (Poedinok et al., 1982).
Izinguquko ze-Cyclic ekwakhekeni kwe-genotypic yabantu emjikelezweni wokuphila we-P. infestans
Umjikelezo wokuthuthuka we-P. infestans endaweni epholile uqukethe izigaba ezi-4.
1) Isigaba sokukhula okunamandla kwesibalo sabantu (isigaba se-polycyclic) ngezizukulwane ezifushane. Lesi sigaba ngokuvamile siqala ngoJulayi futhi sihlala izinyanga ezingu-1,5-2.
2) Isigaba sokumisa ukukhula kwesibalo sabantu ngenxa yokwehla okukhulu kwenani lezicubu ezingathinteki noma ukuqala kwezimo zezulu ezingezinhle. Lesi sigaba emapulazini akhipha amaqabunga ngaphambi kokuvuna siyaphuma emjikelezweni wonyaka.
3) Isigaba sobusika kuma-tubers, esihambisana nokwehla okukhulu kwenani labantu ngenxa yokutheleleka ngephutha kwezilimo eziyizigaxa, ukukhula kancane kokutheleleka kuzo, ukungabikho kokutheleleka kabusha kwezilimo eziyizigaxa, ukubola nokuqothuka kwezilimo eziyizigaxa ezithintekile ngaphansi kwezimo ezijwayelekile zokugcina. .
4) Isigaba sokuthuthukiswa kancane emhlabathini nasezitshalweni (isigaba se-monocyclic), lapho isikhathi sokukhiqiza singafinyelela inyanga noma ngaphezulu (ngasekupheleni kukaMeyi - ekuqaleni kukaJulayi). Ngokuvamile ngalesi sikhathi, amaqabunga anesifo kunzima ukuwabona, ngisho nokubhekwa okukhethekile.
Isigaba sokukhula kwesibalo sabantu abaningi (isigaba se-polycyclic)
Imibono eminingi (Pshedetskaya, Kozubova, 1969; Borisnok, 1969; Osh, 1969; Dyakov, Suprun, 1984; Rybakova, Dyakov, 1990) yabonisa ukuthi ekuqaleni kwe-epiphytoty, i-low virulent futhi i-aggressive kancane, ithathelwa indawo yi-cloves kancane. ezinonya kakhulu futhi ezinolaka, futhi izinga lokukhula kobudlova besibalo sabantu liphakeme, liyancipha ukumelana nezinhlobonhlobo zesitshalo esisingethe.
Njengoba inani labantu likhula, ukuhlangana kwazo zombili izakhi zofuzo ezibalulekile ezikhethwayo ezifakwa ezinhlotsheni zezohwebo (R1-R4) kanye nokungathathi hlangothi ngokukhetha (R5-R11) kuyanda. Ngakho-ke, emiphakathini eseduze neMoscow ngo-1993, isilinganiso sodlame kusukela ngasekupheleni kukaJulayi kuya phakathi no-August senyuke sisuka ku-8,2 saya ku-9,4, futhi ukwanda okukhulu kwaphawulwa ohlotsheni olukhethayo lwe-virulence gene R5 (kusuka ku-31 kuya ku-86% wama-clones anonya) ( Smirnov, 1996).
Ukwehla kwezinga lokukhula kwenani labantu kuhambisana nokuncipha komsebenzi we-parasitic wabantu. Ngakho-ke, eminyakeni yokucindezeleka, kokubili isamba senani lezinhlanga kanye nenani lezinhlanga ezinobudlova kakhulu ziphansi kunezi-epiphytotic (Borisenok, 1969). Uma ekuphakameni kwezimo zezulu ze-epiphytotic zishintsha zibe ezingezinhle ekubhujisweni sekwephuzile futhi ukuhlasela kwamazambane kuncipha, ukugcwala kwama-clones anonya kakhulu futhi anolaka nakho kuyehla (Rybakova et al., 1987).
Ukwanda kwezandiso zofuzo ezithinta udlame kanye nolaka lwesibalo sabantu kungase kube ngenxa yokukhethwa kwama-clones anonya futhi anolaka kubantu abaxubile. Ukukhombisa okukhethiwe, kwasungulwa indlela yokuhlaziya ukuguqulwa kwezakhi zofuzo ezingathathi hlangothi, eyasetshenziswa ngempumelelo kuzibalo ze-chemostat zemvubelo (Adams et al., 1985) kanye ne-Fusarium graminearum (Wiebe et al., 1995).
Imvamisa yezinguquko ezimelana ne-blasticidin S endaweni yabantu be-P. infestans yehla ngokuhambisana nokwanda kolaka lwabantu, okubonisa ushintsho kuma-clones abusayo ngesikhathi sokukhula kwesibalo sabantu (Rybakova et al., 1987) ).
Isigaba sasebusika kuma-tubers
Ngesikhathi sasebusika ezigabeni zamazambane, ubuxhwanguxhwangu kanye nolaka lwezinhlobo ze-P. infestans ziyehla, futhi ukwehla kobugqila kwenzeka kancane kakhulu kunolaka (Rybakova noDyakov, 1990). Ngokusobala, ngaphansi kwezimo ezikhuthaza ukukhula okusheshayo kwesibalo sabantu (ukukhethwa kuka-r), izakhi zofuzo "ezingeziwe" ze-virulence kanye nolaka oluphakeme ziwusizo, ngakho-ke ukuthuthukiswa kwe-epiphytotics kuhambisana nokukhethwa kwama-clones anonya kakhulu futhi anonya. Ngaphansi kwezimo zokugcwala kwendawo ezungezile, lapho kungelona izinga lokuzala, kodwa ukuphikelela kokuba khona ezimeni ezingezinhle (K-ukukhethwa) kuba okubalulekile, izakhi zofuzo "ezingeziwe" zobudlova kanye nolaka zinciphisa ukuqina, futhi ama-clone analezi zakhi zofuzo awokuqala. ukufa, ukuze ulaka olumaphakathi kanye nodlame lwabantu lwehle.
Isigaba sezitshalo emhlabathini
Lesi sigaba siyimfihlakalo kakhulu emjikelezweni wokuphila (Andrivon, 1995). Ukuba khona kwayo kwahlelwa ngokucabangela nje - ngenxa yokuntuleka kolwazi mayelana nokuthi kwenzekani ku-pathogen isikhathi eside (ngezinye izikhathi isikhathi esingaphezu kwenyanga) - kusukela ekuveleni kwezithombo zamazambane kuya ekubonakaleni kwezindawo zokuqala zesifo kuzo. Ngokusekelwe ekuhloleni nasekuhloleni, ukuziphatha kwesikhunta kule nkathi yokuphila kwakhiwe kabusha (uHirst noStedman, 1960; Boguslavskaya, Filippov, 1976).
I-sporulation ye-fungus ingakheka ezigabeni ezinegciwane emhlabathini. Izinhlamvu eziwumphumela zihluma nge-hyphae, ekwazi ukumila isikhathi eside emhlabathini. I-Primary (eyakhelwe ngezilimo eziyizigaxa) neyesibili (emceleni we-mycelium enhlabathini) i-spores ikhuphukela phezu kwenhlabathi ngama-capillary currents, kodwa ithola amandla okuthelela amazambane kuphela ngemva kokuba amaqabunga awo aphansi ehla futhi ehlangana nomhlabathi. Amaqabunga anjalo (okungukuthi, amabala okuqala alesi sifo atholakala kuwo) awakhiwe ngokushesha, kodwa ngemva kokukhula isikhathi eside nokuthuthukiswa kweziqongo zamazambane.
Ngakho-ke, isigaba se-saprotrophic vegetation singase sibe khona emjikelezweni wokuphila we-P. infestans. Uma esigabeni se-parasitic somjikelezo wokuphila ukuhlukumeza kuyisici esibaluleke kakhulu sokufaneleka, khona-ke ekukhethweni kwesigaba se-saprotrophic kuhloswe ukunciphisa izakhiwo ze-parasitic, njengoba kukhonjisiwe ngokuhlolwa kwezinye isikhunta se-phytopathogenic (bheka i-Carson, 1993). Ngakho-ke, kulesi sigaba somjikelezo, izakhiwo ezinolaka kufanele zilahleke kakhulu. Kodwa kuze kube manje akukho ukuhlolwa okuqondile okwenziwe ukuze kuqinisekiswe ukuqagela okungenhla.
Izinguquko zonyaka azithinti kuphela izakhiwo ze-pathogenic ze-P. infestans, kodwa futhi ukumelana ne-fungicides, ekhula esigabeni se-polycyclic (ngesikhathi se-epiphytoties) futhi yehla ngesikhathi sokugcina ubusika (Derevyagina et al., 1991; Kadish noCohen, 1992). Ukwehla okukhulu kokumelana ne-metaxyl kwabonakala phakathi nokutshalwa kwezilimo eziyizigaxa ezithintekile kanye nokuvela kwamachashaza okuqala alesi sifo ensimini.
I-Intraspecific specialization kanye nokuvela kwayo
I-P. infestans idala ubhubhane ezitshalweni ezimbili ezibalulekile kwezentengiso, amazambane kanye notamatisi. Ama-epiphytoties kumazambane aqala ngokushesha ngemva kokuba isikhunta singene ezindaweni ezintsha. Ukunqotshwa kukatamatisi kwaphawulwa ngokushesha ngemva kokuvela kokutheleleka kumazambane, kodwa ama-epiphytoties kutamatisi aphawulwa eminyakeni eyikhulu kamuva - phakathi nekhulu lama-XNUMX. Nakhu okubhalwe nguHallegli noNiederhauser ngokuhlulwa kukatamatisi e-USA
(1962): “Eminyakeni engaba yikhulu ngemva kwe-epiphytosis eqinile ka-100, kwenziwa imizamo embalwa noma cishe kwaze kwatholakala nhlobo yokuthola izinhlobo zikatamatisi ezimelana nezifo. Nakuba i-blight sekwephuzile yaqoshwa okokuqala kumatamatisi emuva ngo-1845, ayizange ibe into yokunaka kakhulu kwabalimi kulesi sitshalo kwaze kwaba yilapho kuqubuka isifo esinamandla ngo-1848. Ensimini yaseRussia, ukubola okuphuzile kukatamatisi kwabhaliswa ngekhulu le-1946. “Sekunesikhathi eside abacwaningi bengasinaki lesi sifo, ngoba asidalanga umonakalo omkhulu kwezomnotho. Kodwa ngo-60s kanye 70s. I-epiphytoties yekhulu lama-XX ye-blight sekwephuzile kutamatisi nayo ibonakala eSoviet Union, ikakhulukazi esifundeni saseLower Volga, e-Ukraine, eNyakatho yeCaucasus, eMoldova ... "(Balashova, 1979).
Kusukela ngaleso sikhathi, ukulimaza utamatisi ngokulimaza sekwephuzile sekube minyaka yonke, kwasakazwa kuyo yonke indawo yezimboni nezasekhaya futhi kubangela umonakalo omkhulu wezomnotho kulesi sitshalo. Kwenzenjani? Kungani ukubonakala kokuqala kwe-parasite kumazambane kanye ne-epiphytotic lesion yalesi siko kwenzeka cishe ngasikhathi sinye, futhi kungani kwathatha ikhulu leminyaka ukuthi i-epiphytotic ivele kutamatisi? Lo mehluko usekela umuntu waseMexico esikhundleni somthombo waseNingizimu Melika wokutheleleka. Uma izinhlobo ze-Phytophthora infestans zakhula njenge-parasite yezinhlobo ezithwala izigaxa zaseMexico zohlobo lwe-Solanum, khona-ke kuyaqondakala ukuthi kungani amazambane atshaliwe ayingxenye yohlobo olufanayo nezinhlobo zaseMexico athinteke kakhulu, kodwa ngenxa yokungabikho kwamazambane. Ukuziphendukela kwemvelo ngokubambisana ne-parasite, engazange yakhe izindlela zokumelana okuqondile nokungaqondile.
Utamatisi ungowesigaba esihlukile sohlobo, uhlobo lokushintshisana kwalo lunomehluko omkhulu kusuka ezinhlobonhlobo ze-tuberous, ngakho-ke, naphezu kokuthi utamatisi awukho ngaphandle kokukhethekile kokudla kwe-P. infestans, ukuqina kokunqotshwa kwawo kwakunganele. ngenxa yokulahlekelwa okukhulu kwezomnotho.
Ukuvela kwama-epiphytoties kutamatisi kungenxa yezinguquko ezinkulu zofuzo ku-parasite, okwandisa ukuguquguquka kwayo (i-pathogenicity) lapho i-parasite. Sikholelwa ukuthi uhlobo olusha, olukhethekile lokufakela i-parasitizing utamatisi umjaho we-T1 ochazwe ngu-M. Gallegly, othinta izinhlobo zikatamatisi we-cherry (Red Cherry, Ottawa), ukumelana nomjaho we-T0 owandile kumazambane (Gallegly, 1952). Ngokusobala, uguquko (noma uchungechunge lwezinguquko) oluguqule umjaho we-T0 emjahweni we-T1 futhi lwaholela ekuveleni kwama-clones ajwayelane kakhulu nokunqotshwa kukatamatisi. Njengoba kuvame ukwenzeka, ukwanda kwe-pathogenicity kumsingathi oyedwa kuhambisane nokwehla kuye komunye, okungukuthi, kwavela ubuchwepheshe bokuqala, obungakabikho obuphelele be-intraspecific - kumazambane (umjaho T0) nakutamatisi (umjaho T1).
Yibuphi ubufakazi obusekela lo mbono?
- Ukwenzeka kumazambane notamatisi. Emaqabungeni katamatisi, umjaho we-T1 uyagqama, kanti emaqabunga amazambane awuvamile. Ngokusho kuka-S.F.Bagirova kanye no-T.A. I-Oreshonkova (engashicilelwe) esifundeni saseMoscow ngo-1991-1992, ukuvela komjaho we-T1 ekutshalweni kwamazambane kwakungu-0%, kanti ekutshalweni kukatamatisi - 100%; ngo-1993-1995 - 33% no-90%, ngokulandelana; ngo-2001 - 0% kanye 67%. Idatha efanayo yatholwa kwa-Israel (Cohen, 2002). Ukuhlolwa kokutheleleka kwezilimo eziyizigaxa zamazambane ezinama-isolate omjaho we-T1 kanye nengxube ye-isolate T0 kanye ne-T1 kubonise ukuthi i-isolate yomjaho we-T1 ayigcinwanga kahle kuma-tubers futhi ithathelwa indawo yi-isolate yomjaho we-T0 (Dyakov et al., 1975; Rybakova, 1988).
2) Amandla omjaho we-T1 ekutshalweni kukatamatisi. Ukutheleleka okuyinhloko kwamaqabunga katamatisi kwenziwa yi-isolate yomjaho we-T0, ophethe ekuhlaziyeni ukutheleleka ezindaweni zokuqala ezakhiwe emaqabunga. Lokhu kuqinisekisa uhlelo olwamukelwayo jikelele lokufuduka kwezimuncagazi: Isibalo esikhulu sokutheleleka okuvela kumazambane senziwa umjaho we-T0, nokho, inani elincane lama-T1 clones agcinwe kumazambane, kanye kutamatisi, asusa umjaho we-T0 futhi anqwabelane ngasemgwaqweni. ukuphela kwesikhathi se-epiphytotic. Kungenzeka futhi ukuthi kukhona omunye umthombo wokutheleleka kwamaqabunga katamatisi ngomjaho we-T1, ongenamandla njengezilimo eziyizigaxa zamazambane namaqabunga, kodwa njalo. Ngakho-ke, lo mthombo unethonya elincane esakhiweni sofuzo sabantu abathelela utamatisi, kodwa kamuva unquma ukuqoqwa komjaho we-T1 (Rybakova, 1988; Dyakov et al., 1994).
3) Ukuba nolaka kumazambane notamatisi. Ukutheleleka okwenziwa ngamahlamvu katamatisi namazambane anezinhlanga ezihlukene i-T0 ne-T1 kubonise ukuthi okokuqala kunolaka kakhulu kumazambane kunotamatisi, kanti lezi zamuva zinolaka kakhulu kutamatisi kunamazambane. Lo mehluko ubonakala ekususweni kwabantu abangabodwana bezinhlanga ezingezona “ezingowabo” besuka kubantu abaxubile ngesikhathi sokudlula emaqabungeni endaweni ebamba ukushisa (Dyakov et al., 1975) nasezindaweni zasensimini (Leberton et al., 1999); umehluko emthwalweni omncane othathelwanayo, isikhathi se-latency, ubukhulu bezindawo ezithathelwanayo kanye nokukhiqizwa kwezinhlamvu (Rybakova, 1988; Dyakov et al., 1994; Legard et al., 1995; Forbes et al., 1997; Oyarzun et al., 1998; Leberton et al., 1999; Vega-Sanchez et al., 2000; Knapova, Gisi, 2002; Sussuna et al., 2004).
Ulaka lwama-isolates omjaho we-T1 kuya ezinhlobonhlobo zikatamatisi ezintula izakhi zofuzo ziphezulu kangangokuba lezi zihlukanisi zikhula emaqabunga njengasendaweni yezondlamzimba ngaphandle kokwenza izicubu ezithelelekile (Dyakov et al., 1975; Vega-Sanchez et al., 2000) .
4) I-Virulence yamazambane notamatisi. Umjaho we-T1 uthinta izinhlobo ze-cherry tomato gene ukumelana ne-Ph1, kanti umjaho we-T0 awukwazi ukuthelela lezi zinhlobonhlobo, i.e. ine-virulence encane. Mayelana nabahlukanisi
I-R-genes yamazambane ihlobene ngokuphambene, i.e. izinhlobo ezihlukanisiwe namaqabunga katamatisi azinabungozi kakhulu kunezinhlobo "zamazambane" (Ithebula 11).
5) Omaka abangathathi hlangothi. Ukuhlaziywa komaka abangathathi hlangothi kuzibalo ze-P. infestans ezidla amazambane notamatisi nakho kufakazela ukukhethwa kwe-intraspecific ye-multidirectional. Emphakathini waseBrazil we-P. infestans, i-tomato leaf isolate yayiyingxenye yomugqa we-clonal US-1, kanti lawo avela emaqabungeni amazambane ayengomugqa we-BR-1 (Suassuna et al., 2004). EFlorida (USA), kusukela ngo-1994, i-clone US-90 yaqala ukubusa kumazambane (ngesenzakalo esingaphezu kuka-8%), futhi ihlanganisa i-US-11 ne-US-17 kutamatisi, kanti lezi zindawo zokugcina zinolaka kakhulu kutamatisi. kunamazambane (Weingartner, Tombolato, 2004). Umehluko omkhulu kumafrikhwensi e-genotype (izigxivizo zeminwe ze-DNA) kuma-isolate amazambane kanye notamatisi wasungulwa ku-1200 P. infestans strains eqoqwe e-United States kusukela ngo-1989 kuya ku-1995 (Deahl et al., 1995).
Ukusebenzisa indlela ye-AFLP kwenza kwaba nokwenzeka ukuhlukanisa izinhlobo ezingama-74 eziqoqwe emaqabunga amazambane namatamatisi ngo-1996-1997. eFrance naseSwitzerland, baba ngamaqembu angu-7. Izinhlobo zamazambane notamatisi azizange zihluke ngokusobala, kepha izinhlobo "zamazambane" zihluke kakhulu kunofuzo "zamazambane". Owokuqala atholwe kuwo wonke amaqoqo ayisikhombisa, kanti lena yokugcina, kwamane kuphela, okubonisa i-genome ekhethekile kakhulu yakamuva (Knapova kanye noGisi, 2002).
6) Izindlela zokuzihlukanisa. Uma isibalo se-parasite ezinhlotsheni ezimbili zezitshalo ezisingathayo siguqukela ekunciphiseni ukugxila ekusingatheni "ezazo", khona-ke izindlela ezihlukahlukene zangaphambi kwesikhathi nezangemva kokuphila kwesikhathi ziyavela ezivimbela ukushintshaniswa kwezakhi zofuzo (Dyakov kanye noLekomtseva, 1984).
Ucwaningo oluningana luphenye umphumela womthombo wezinkinga zabazali ekusebenzeni kahle kwe-hybridization. Lapho kuwela izinhlobo ezihlukanisiwe nezinhlobo ezahlukene zohlobo lwe-Solanum e-Ecuador (u-Oliva et al., 2002), kwatholakala ukuthi izinhlobo ezinohlobo lokuhlangana kwe-A2 oluvela endle iSolanaceae (umugqa we-clonal EC-2) weqa okubi kakhulu ngezinhlobo zikatamatisi ( umugqa EC -3), futhi wawela ngempumelelo kakhulu uhlobo lwamazambane (EC-1).
Wonke ama-hybrids atholakale engena-pathogenic. Ababhali bakholelwa ukuthi iphesenti eliphansi le-hybridization kanye nokuncipha kwe-pathogenicity kuma-hybrids kungenxa yezinqubo ze-postmeiotic zokuhlukaniswa kwabantu ngokuzala.
Ekuhlolweni kuka-Bagirova et al. (1998), inqwaba yezinhlobo zamazambane notamatisi zawela nezakhiwo zemijaho ye-T0 ne-T1. Iziphambano ezivunde kakhulu zezinhlobo ze-T1xT1 ezihlukanisiwe kutamatisi (ama-oospores angama-36 endaweni yokubuka yesibonakhulu, u-44% wokuhluma kwe-oospore), ezingasebenzi kahle kube yiziphambano zezinhlanga ze-T0xT1 ezihlukanisiwe kubasingathi abahlukahlukene (inani eliphansi lama-oospores asathuthuka futhi ahluma, ingxenye enkulu yama-oospores akhipha izisu futhi angathuthukisiwe) ... Ukusebenza kahle kweziphambano phakathi kwama-isolate omjaho we-T0 ohlukaniswe namazambane kuvele kwaba okumaphakathi. Njengoba umzimba omkhulu wezinhlobo zomjaho we-T0 uthinta amazambane, unomthombo onokwethenjelwa wobusika - izilimo eziyizigaxa zamazambane, ngenxa yalokho ukubaluleka kwama-oospores njengamayunithi athathelwanayo ebusika kubantu abavela kumazambane aphansi. "Ifomu likatamatisi" eliguquliwe liyakwazi ukuhlala ebusika kutamatisi ngendlela ye-oospores (bheka ngezansi) ngakho-ke igcina ukukhiqiza okuphezulu kwenqubo yocansi. Ngenxa yokuzala kwayo okuphezulu, i-T1 ithola amandla azimele okutheleleka okuyinhloko kutamatisi. Imiphumela etholwe ngu-Knapova et al. (Knapova et al., 2002) ingahunyushwa ngendlela efanayo. Iziphambano zezinhlobo ezihlukanisiwe namazambane anezinhlobo ezivela kutamatisi zinikeze inani eliphakeme kakhulu lama-oospores - 13,8 per sq. Mm. medium (ngokusabalala kwe-5-19) kanye nephesenti eliphakathi lokuhluma kwama-oospores (6,3 ngokusabalala kwe-0-24). Ukuphambana kwezinhlobo ezihlukanisiwe kutamatisi kunikeza iphesenti eliphansi kakhulu lama-oospores (7,6 ngokusabalala kuka-4-12) ngephesenti eliphakeme kakhulu lokuhluma kwawo (10,8). Iziphambano phakathi kwezinhlobo ezihlukanisiwe kumazambane zinikeze inani eliphakathi lama-oospores (8,6 ngokuhlakazeka okuphezulu kwedatha - 0-30) kanye nephesenti eliphansi kakhulu lokuhluma kwama-oospores (2,7). Ngakho-ke, izinhlobo zamazambane azivundile kakhulu kunalezo eziphuma kutamatisi, kodwa iziphambano zokuhlanganisana kwabantu azizange zinikeze imiphumela emibi kakhulu kune-intrapopulation. Kungenzeka ukuthi umehluko ngedatha engenhla ngu-Bagirova et al. kuchazwa iqiniso lokuthi abacwaningi baseRussia basebenze nezinkinga ezihlukanisiwe ekuqaleni kweminyaka engama-90 yekhulu lamashumi amabili, nabacwaningi baseSwitzerland - abanezinkinga ezihlukanisiwe ngasekupheleni kwawo-90s.
Ukuzala okuphansi kungase kube ngenxa yezinhlobo ze-heteroploid. Uma emiphakathini yaseMexico, lapho inqubo yocansi kanye nokutheleleka okuyinhloko nge-oospore nzalo kuvamile, iningi lezinhlobo ezifundwayo ze-P. infestans ziyi-diploid, khona-ke emazweni ase-Old World intrapopulation polymorphism ye-ploidy iyabonwa (di-, tri- kanye nezinhlobo ze-tetraploid, kanye nezinhlobo ze-heterokaryotic ezine-heteroploid nuclei) , nezinkinga ezinezinhlobo ezahlukene zokuhlangana, i.e. evundile ngokulinganayo, ihluke ku-nuclear ploidy (Therrien et al., 1989, 1990; Whittaker et al., 1992; Ritch, Daggett, 1995). Ukuhlukahluka kwama-nuclei ku-antheridia kanye ne-oogonia kungabangela ukuzala okuphansi.
Ngokuphathelene nokuhwebelana ngezikhali zenuzi phakathi kwe-hyphae ngesikhathi se-anastomoses, lokhu kuvinjwa ukungahambelani kwezitshalo, okuhlukanisa abantu abangasexual babe ama-clones amaningi angawodwa ngofuzo (Poedinok noDyakov, 1987; Gorbunova et al., 1989; Anikina et al., 1997b).
7) Ukuhlangana kwabantu. Idatha engenhla ibonisa ukuthi ukuxutshwa phakathi kwezinhlobo "zamazambane" kanye "notamatisi" we-P. infestans kungenzeka. Ukutheleleka kabusha okuhambisanayo kwabasingathi abahlukahlukene nakho kungenzeka, nakuba ngolaka oluncishisiwe.
Ucwaningo lwezimpawu zenani labantu ezindaweni ezingazodwa ezivela emasimini amazambane kanye notamatisi aseduze ngo-1993 lwabonisa ukuthi cishe ingxenye yesine yamahlamvu ahlukanisiwe namahlamvu katamatisi yasuswa ensimini yamazambane engumakhelwane (Dolgova et al., 1997). Ngokwethiyori, kungacatshangwa ukuthi ukwehlukana kwenani labantu kumahostela amabili kuzokwanda futhi kuholele ekuveleni kwezinhlobo ezikhethekile ze-intraspecific (f.sp. potato kanye notamatisi we-f.sp), ikakhulukazi njengoba ama-oospores angaqhubeka emfucumfucu yezitshalo (Drenth et al., 1995; Bagirova, Dyakov, 1998) kanye nembewu yetamatisi (Rubin et al., 2001). Ngenxa yalokho, utamatisi okwamanje unomthombo wokuvuselelwa kwentwasahlobo ngaphandle kwezilimo eziyizigaxa zamazambane.
Nokho, konke kwenzeka ngendlela ehlukile. Ukucwiliswa ngokweqile kwe-oospores kwavumela i-parasite ukuthi igweme isigaba esincane kunazo zonke emjikelezweni wayo wokuphila - isigaba se-monocyclic sezitshalo emhlabathini, lapho izakhiwo ze-parasitic ziyancipha, ezibuyiselwa kancane kancane esigabeni se-polycyclic ehlobo.
Ithebula 11. Ukuvama kwezakhi zofuzo ze-virulence ezinhlobonhlobo zokuhlukanisa amazambane ezinhlotsheni ze-P. infestans
izwe | I-Год | Isilinganiso senani lezakhi zofuzo ze-virulence ezinhlotsheni | umbhali | |
kusukela amazambane | kusuka kutamatisi | |||
France | 1995 | 4.4 | 3.3 | Leberton et al., 1999 |
1996 | 4.8 | 3.6 | Leberton, Andrivon, 1998 | |
France, Switzerland | 1996-97 | 6.8 | 2.9 | Knapova, Gisi, 2002 |
США | 1989-94 | 5 | 4.8 | Goodwin et al., 1995 |
USA, Zap. Washington | 1996 | 4.6 | 5 | UDorrance et al., 1999 |
1997 | 6.3 | 3.5 | " | |
Ecuador | 1993-95 | 7.1 | 1.3 | Oyarzun et al., 1998 |
Израиль | 1998 | 7 | 4.8 | Cohen, 2002 |
1999 | 6 | 5.7 | " | |
2000 | 6.7 | 6.1 | " | |
Russia, Mosk. isifunda | 1993 | 8.9 | 6.7 | USmirnov, ngo-1996 |
Russia, izifunda ezahlukene | 1995 | 9.4 | 8 | Kozlovskaya nabanye. |
1997 | 9.2 | 9.2 | " | |
2000 | 8.7 | 4.8 | " |
I-zoosporangia eyisisekelo kanye nama-zoospores, ahluma ama-oospores, anezinga eliphezulu lomsebenzi we-parasitic, ikakhulukazi uma ama-oospores akhiwa i-parthenogenetically ngaphansi kwethonya lama-pheromones ohlobo oluphambene nohlobo lokukhwelana. Ngakho-ke, izinto ezithathelwanayo ezitshalweni zikatamatisi ezitshalwe embewini etheleleke nge-oospores ziyingozi kakhulu kukho kokubili utamatisi namazambane.
Lezi zinguquko ziholele ekuhlelweni kabusha kwesibalo sabantu, okuvezwe kulezi zinguquko ezibalulekile ezilandelayo ngokombono we-epidemiological:
- Izithombo ze-tomato ezinegciwane ziye zaba umthombo obalulekile wokutheleleka okuyinhloko kwamazambane (uFilippov, i-Ivanyuk, imilayezo yomuntu siqu).
- Ama-epiphytoties kumazambane aqala ukubonwa ekuqaleni kukaJuni, cishe inyanga ngaphambi kokujwayelekile.
- Ekutshalweni kwamazambane, amaphesenti omjaho we-T1 anyukile, okwakuhlangatshezwane nawo lapho ngenani elincane (Ulanova et al., 2003).
- Izinhlobo ezihlukanisiwe namahlamvu katamatisi azisahlukanga ezinhlamvini zamazambane ngokuya ngobungozi bokuhlukanisa amazambane ezakhi zofuzo ze-virulence futhi zaqala ukudlula izinhlobo "zamazambane" ngobudlova hhayi kutamatisi kuphela, kodwa nakumazambane (Lavrova et al., 2003; Ulanova et. al., 2003).
Ngakho-ke, esikhundleni sokuhlukana, kwaba khona ukuhlangana kwabantu, ukuvela kwenani labantu elilodwa ezitshalweni ezimbili ezibambayo ezinobulwane obuphezulu kanye nolaka kuzo zombili izinhlobo.
isiphetho
Ngakho-ke, naphezu kweminyaka engaphezu kwe-150 yocwaningo olunzulu lwe-P. infestans, ku-biology, kuhlanganise ne-biology yabantu balesi sitho esiyimbangela yezifo ezibaluleke kakhulu zezitshalo ze-solanaceous ezitshaliwe, kuningi okungaziwa. Akukacaci ukuthi ukudlula kwezigaba ngazinye zomjikelezo wokuphila kusithinta kanjani ukwakheka kwabantu, yiziphi izindlela zofuzo zokuguquguquka kwe-canalized of aggressiveness and virulence, siyini isilinganiso sezinhlelo zokuzala kanye ne-clonal zokuzala kubantu bemvelo, kanjani ukungezwani vegetative ifa, iyini indima amazambane kanye utamatisi in the ukutheleleka primary yalezi zitshalo futhi kuyini umphumela wabo phezu isakhiwo izimuncagazi labantu. Kuze kube manje, izindaba ezibalulekile ezibalulekile njengezinqubo zofuzo zokushintsha ulaka lwe-parasite noma ukuguguleka kokumelana namazambane okungaqondile azikaxazululwa. Ngokujula nokwanda kocwaningo mayelana nokulimaza kwamazambane sekwephuzile, i-parasite ibeka izinselelo ezintsha kubacwaningi. Kodwa-ke, ukuthuthukiswa kwamakhono okuhlola, ukuvela kwezindlela ezintsha ze-methodological zokukhohlisa ngezakhi zofuzo namaprotheni kusivumela ukuba sibe nethemba lesixazululo esiyimpumelelo semibuzo ebuziwe.
Isihloko sashicilelwa kujenali "Potato Protection" (No. 3, 2017)