D. Yu. Ryazantsev, E. M. Chudinova, L. Yu. Kokaeva, S. N. Elansky, P. N. Balabko, G. L. Belova, S. K. Zavriev
I-Phytopathogenic fungus i-Colletotrichum coccodes idala izifo eziyingozi kumazambane kanye notamatisi owaziwa ngokuthi i-anthracnose kanye ne-black spot of tubers. Ngezici ze-morphological, ngokuvamile kunzima ukuhlukanisa nezifo ezibangelwa amanye ama-microorganisms; ngezithelo zikatamatisi oluhlaza, lesi sifo singaba yi-asymptomatic, sizibonakalisa kuphela ezithelweni ezibomvu ezivuthiwe. Ukuze uthole ukuxilongwa okusheshayo nokunembile kwe-pathogen, isistimu yokuhlola ye-PCR yesikhathi sangempela inikezwa. Ukuze kuthuthukiswe uhlelo lokuhlola, kwanqunywa ukulandelana kwe-nucleotide yofuzo lwe-glycerol triphosphate dehydrogenase engu-45 C. ama-coccodes ahlukaniswe nezilimo eziyizigaxa zamazambane ezifundeni ezahlukene zaseRussia.
Ngokusekelwe emiphumeleni etholiwe kanye nokuhlaziywa kokulandelana okufanayo kwezinye izinhlobo zezilwane ezitholakala kusizindalwazi se-GenBank, ama-primers aqondene nezinhlobo ezithile kanye ne-probe yama-C. coccodes aklanywa. Ukuhlola ukucaciswa kwesistimu yokuhlola edaliwe, i-PCR yenziwa nge-DNA ehlukanisiwe namasiko ahlanzekile ezinhlobo ezingu-15 ezihlukene zesikhunta se-parasitic kanye ne-saprotrophic ehambisana nezitshalo zikatamatisi namazambane (Fusarium oxysporum, F. verticillium, Phomopsis phaseoli, Alternaria alternata, Helminthosporium solani , Colletotrichum coccodes Phellinus ferrugineovelutinus, Stemphylium vesicarium, Helminthosporium solani, Phomopsis phaseoli, Neonectria radicicola, Rhizoctonia solani, Penicillium sp., Cladosporium fulvum, C. cladosporioides). Ukuba khona kwe-Colletotrichum coccodes DNA kwanqunywa kumjikelezo we-threshold wama-20-27, kuyilapho ezinye izinhlobo zatholwa ngemva kwemijikelezo engu-40 noma zingabonwa. Isistimu yokuhlola yenza kube nokwenzeka ukuthola ngokuthembekile ukugxila kwe-C. coccodes DNA engaphezu kuka-0.01 ng/mm3 kungxube ye-PCR ehlaziyiwe. Ngokusebenzisa uhlelo lokuhlola oluthuthukisiwe, ukuba khona kwe-C. cocodes emaqabunga katamatisi anezimpawu zezifo zokukhunta kanye nezilimo eziyizigaxa zamazambane ngaphandle kwezimpawu zangaphandle zesifo kwaphenywa. Amaqabunga anezimpawu zokutheleleka kwefungal aqoqwe emikhakheni emibili ehlukene yaseKrasnodar Territory, izilimo eziyizigaxa - ezivela emasimini aseKostroma, eMoscow, eKaluga, eNizhny Novgorod. Iqabunga elilodwa likatamatisi eliqukethe i-C. coccodes DNA latholakala eKrasnodar Territory; ukutholakala okuphawulekayo kwe-DNA yale pathogen kutholwe kumasampula angu-5 wezilimo eziyizigaxa ezitshalwe ezindaweni zaseKostroma, eMoscow, eKaluga.
Isingeniso
Amakhowe ohlobo lwe-Colletotrichum angama-phytopathogens ayingozi athinta okusanhlamvu, imifino, amakhambi, izithelo ezingapheli kanye nezitshalo zamajikijolo. Enye yezinhlobo ezitholakala yonke indawo yalolu hlobo, i-Colletotrichum coccodes (Wallr).
I-Hughes, iyimbangela ye-anthracnose nendawo emnyama yamazambane notamatisi, futhi ibangela izifo zenani lezinye izitshalo zomndeni wakwaSolanaceae, kuhlanganisa. ukhula (Dillard, 1992). I-C. cocodes ithinta zonke izingxenye ezingaphansi komhlaba zesitshalo, izisekelo zesiqu, amaqabunga, nezithelo (Andrivon et al., 1998; Johnson, 1994). Ekhasini lezilimo eziyizigaxa zamazambane ezinegciwane, ukuthuthukiswa kwamabala ampunga anemiphetho engasho lutho, lapho kubonakala khona amachashazi amnyama we-sporulation kanye ne-microsclerotia. Ngesikhathi sokugcina, izilonda ezinokuqukethwe okuthambile zingakha ku-pulp yezilimo eziyizigaxa, i.e. lesi sifo singena esigabeni se-anthracnose, okuyinto, nokho, engavamile kakhulu.
Ngesikhathi esifanayo, izimpawu ze-anthracnose (izilonda zesikhumba ezinamachashazi amancane amnyama) zifana nezithelo zikatamatisi. Emaqabungeni, izimpawu ze-C. cocodes zivela njengamachashaza ansundu ngokumnyama, ngokuvamile azungezwe izicubu eziphuzi (Johnson, 1994).
Ukuthuthukiswa kwendawo emnyama kuma-tubers konakalisa ukubukeka kwawo, okugqama kakhulu lapho kuthengiswa amazambane abomvu ahlanjululwe. I-Peel delamination iholela ekuhwamukeni okweqile kanye nokwanda kokulahleka kwesitoreji (Indlala noMcIntyre, 1979). Ukulimala kwezinye izitho zezitshalo kuholela ekulahlekelweni kwesivuno, okuphawulwe endaweni evulekile nevaliwe (Johnson, 1994; Tsror et al., 1999). Izifo ezibangelwa ama-C. cocodes zivamile cishe kuzo zonke izifunda zomhlaba ezikhiqiza amazambane, kuhlanganise neRussia (Leesa, Hilton, 2003; Belov et al, 2018). Ukulawulwa kwalezi zifo kunzima ngenxa yokungasebenzi kahle kwemithi ebulala isikhunta ekhona ngokumelene ne-C. cocodes kanye nokuntuleka kwezinhlobo ezikwazi ukumelana (Funda, Fihla, 1995).
I-inoculum ye-C. coccodes ingaqhubeka ezigabeni zembewu (Funda, Fihla, 1988; Johnson et al., 1997), imbewu katamatisi (Ben-Daniel et al., 2010), iphila isikhathi eside emhlabathini, emfucumfucwini yezitshalo. (Dillard, 1990; Dillard, Cobb, 1993) kanye nokhula (Raid, Pennypacker, 1987). Imisebenzi yababhali abaningana (Funda, Fihla, 1988; Barkdoll, Davis, 1992; Johnson et al., 1997; Dillard, Cobb, 1993) ibonise ukuthi ukukhula kwalesi sifo kumazambane notamatisi kuncike kakhulu ebukhoneni. ye-inoculum embewini nasenhlabathini. Ngakho-ke, ukuze kuncishiswe ukulahlekelwa okuvela kulesi sifo, kuyadingeka ukuxilonga (kuhlanganise nobuningi) ama-propagules we-fungus entweni yembewu, enhlabathini, kumbewu yamazambane nezilimo eziyizigaxa kanye nembewu katamatisi ebekwe ukugcinwa. Ukuxilongwa kwe-morphological enhlabathini nasezitshalweni kungenziwa kuphela ngokuba khona kwe-microsclerotia, kodwa-ke, etholakala kwezinye izinhlobo zesikhunta.
Izimpawu ezikulezi zimila zifana kakhulu notwayi oluyisiliva olubangelwa ukhunta iHelminthosporium solani. Ukuhlukaniswa kwe-Colletotrichum coccodes kanye ne-Helminthosporium solani ibe isiko elihlanzekile kulula futhi kuthatha isikhathi eside ngenxa yokukhula kancane endaweni yezakhi. Ukuze uhlonze ngokushesha ama-Colletotrichum coccodes, kuyadingeka ukusebenzisa izindlela zokuxilonga eziwusizo. Indlela elula kunazo zonke i-polymerase chain reaction (PCR) nokuguqulwa kwayo - i-PCR yesikhathi sangempela. Njengamanje, uhlelo lokuhlola olwakhiwe abacwaningi baseBrithani (uCullen et al., 2002) lwesifunda se-ITS1 se-rDNA lusetshenziswa eYurophu nase-United States. Ukusetshenziswa kwayo kubonise imiphumela emihle ekuhlaziyweni kwama-isolate aseRussia (Belov et al, 2018). Kodwa-ke, i-C. coccodes iyahlukahluka kakhulu futhi ukutholakala kwayo ngokulandelana kwe-DNA eyodwa kungaholela emiphumeleni engemihle engamanga. Ukuze uthole ukuxilongwa okunokwethenjelwa, kuyadingeka ukuhlaziya ukulandelana kwe-DNA kwezinhlobo ezithile zezinhlobo ezithile, mayelana nalo esiye sakha uhlelo lokuhlola lwangempela oluvumela ukuhlonza ama-C. coccodes ngokulandelana kofuzo lwe-glyceraldehyde-3-phosphate dehydrogenase.
Izimpahla nezindlela
Ukuhlola ukusebenza kahle kanye nokucaciswa kwezinhlelo zokuhlola ezidaliwe, sisebenzise amasiko amsulwa ezinhlobo eziyi-15 zesikhunta ezihlukaniswe ababhali kumasampula agulayo amaqabunga katamatisi nezithelo, izilimo eziyizigaxa zamazambane (Ithebula 1). Ukuze sizihlukanise, sathatha izitho zezitshalo ezinezimpawu zokutheleleka kwefungal, hhayi isitho esingaphezu kwesisodwa esihlahleni ngasinye.
Ucezu lwe-tuber enekhasi, ucezu lwesithelo sikatamatisi, neqabunga elithintekile kwafakwa ngaphansi kwesibonakhulu se-binocular, okwathi ngemva kwalokho i-mycelium, izinhlamvu, noma ucezu lwethishu lwadluliselwa ku-agar medium (wort agar) endaweni. Isitsha sikaPetri esinenaliti yokuhlinza ebukhali ebukhali. Ama-isolate agcinwe ku-agar slant kumashubhu okuhlola ku-4 ° C.
Amasampula amaqabunga e-tomato anezimpawu zezifo zokukhunta, okuhloswe ukuhlaziya, ngokushesha ngemva kokuqoqwa (ensimini) afakwa ku-70% we-ethyl alcohol lapho agcinwa khona kuze kuhlukaniswe i-DNA. Izilimo eziyizigaxa zamazambane zilethwa elabhorethri, zihlutshiwe (2 × 1 cm ucezu) kuzo, futhi ziqandiswe ku-20 ° С. Igcinwe ifriziwe kuze kube yilapho i-DNA ihlukaniswa.
Amasiko ahlanzekile esikhunta sokuhlukaniswa kwe-DNA ayetshalwa endaweni ye-liquid pea medium. I-mycelium yesikhunta isusiwe endaweni ewuketshezi, yomiswa ephepheni lokuhlunga, iqandiswe ku-nitrogen ewuketshezi, ifakwe i-homogenized, ifakwe ku-CTAB buffer, ihlanzwe nge-chloroform, igcwaliswe ngengxube ye-isopropanol ne-0.5 M potassium acetate, igezwe kabili nge-2% utshwala. I-DNA ewumphumela yachithwa emanzini angcolile futhi yagcinwa ku--70 ° С (Kutuzova et al., 20). Ukugxiliswa kwe-DNA kukalwa kusetshenziswa ikhithi yokulinganisa ye-HS DNA ye-DNA emuke kabili ku-Qubit 2017 (Qiagen, Germany). Amasampula otshwala naqandisiwe afakwa ku-nitrogen ewuketshezi, kwase kuthi ukukhishwa kwe-DNA kwenziwa njengoba kuchazwe ngenhla (ku-mycelium yamasiko esikhunta ahlanzekile).
Ithebula 1. Umsuka wezinhlobo zesikhunta ezisetshenziswa emsebenzini
Igama lamakhowe | Isitshalo, isitho | Indawo yokukhetha |
---|---|---|
Colletotrichum coccodes 1, C. coccodes 2, C. coccodes 3, Ilyonectria crassa, Rhizoctonia solani | amazambane tuber | Isifunda saseKostroma, izilimo eziyizigaxa zamazambane zesizukulwane sokuqala sensimu, i-cultivar Red Scarlett |
Amakhodi we-Colletotrichum 4 | iqabunga lamazambane | Ummeli. Mari El, Yoshkar-Ola |
I-Helminthosporium solani | amazambane tuber | Isifunda saseMagadan, pos. Itende, i-tuber yamazambane |
I-Cladosporium fulvum | iqabunga likatamatisi | Isifunda saseMoscow, utamatisi onezithelo ezinkulu |
I-Alternaria tomatophila | isithelo sikatamatisi | inikezwe abasebenzi belabhorethri ye-mycology kanye ne-phytopathology ye-All-Russian Research Institute of Plant Protection |
Fusarium verticillium, Phomopsisphaseoli, Alternaria alternata, Phellinus ferrugineovelutinus, Stemphylium vesicarium, Cladosporium cladosporioides, Acrodontium luzulae, Penicillium sp. | isithelo sikatamatisi | I-Krasnodar Territory, isifunda saseCrimea, i-grade Cream |
I-Fusarium oxysporum | impande kakolweni | Isifunda saseMoscow |
I-PCR yenziwa nge-DTprime amplifier (DNA-Technology). Ku-PCR, iziqalo zangempela kanye nophenyo lwesifunda esithile sezinhlobo zofuzo lwe-glycerol triphosphate dehydrogenase kwasetshenziswa: i-forward primer Coc70gdf –TCATGATATCATTTCTCTCACGGCA, i-primer ehlanekezelayo Coc280gdr - TACTTGAGCATGTAGGCCTGGCGGGT1 - FACTTTAGTAGGGGGT213 proprimer. Ama-primers akhulisa isifunda esingu-XNUMX bp.
Ukusabela kuthathe ama-50 ng engqikithi ye-DNA (lapho kuhlaziywa amaqabunga nezilimo eziyizigaxa) kanye no-10 ng (lapho kuhlaziywa i-DNA kumasiko amsulwa esikhunta). Ingxube yokusabela (35 μl) yahlukaniswa ungqimba lukapharafini lwaba izingxenye ezimbili: ephansi (20 μl) iqukethe u-2 μl we-10 × i-reaction buffer (750 mM Tris-HCl, pH 8.8; 200 mM (NH4) 2SO4; 25 mM MgCl2; 0.1% Tween- 20), 0.5 mM we-deoxynucleotide triphosphate ngayinye, 7 pmol we-primer ngayinye, kanye no-4 pmol weprobe ye-hydrolyzable fluorescent; engaphezulu ibiqukethe i-1 μl ye-10 × PCR buffer kanye ne-1 U ye-Taq polymerase.
Ukuhlukaniswa kwengxube nopharafini kuvumela amashubhu ukuthi agcinwe isikhathi eside ezingeni lokushisa elingu-5 ° C futhi anikeze isiqalo esishisayo se-PCR ngemva kokuwashisa imizuzu eyi-10 ekushiseni okungaphezu kuka-80 ° C. I-PCR yenziwa ngokohlelo olulandelayo: 94.0 ° C - 90 s (umjikelezo ongu-1); 94.0 ° C - 30 s; 64.0 ° C - 15 s (5 imijikelezo); 94.0 ° C - 10 s; 64.0 ° C - 15 s (45 imijikelezo); 10.0 ° C - isitoreji.
Imiphumela nengxoxo
Ukulandelana kofuzo lwe-glycerol triphosphate dehydrogenase kwanqunywa ezinhlotsheni ezingama-45 ezihlukaniswe namaqabunga, iziqu, izilimo eziyizigaxa zamazambane kanye nezithelo zikatamatisi (Kutuzova, 2018) ezindaweni ezahlukahlukene zaseRussia. Ukulandelana okuphenyiwe kwazo zonke izinhlobo kuhlukaniswe ngamaqembu ama-2 ahlukile kuma-nucleotide amabili. Ukulandelana kwe-nucleotide yabameleli bawo womabili amaqembu ngaphansi kwezinombolo KY496634 kanye ne-KY496635 kufakwe ku-GenBank.
Ama-primers i-coc70gdf, coc280gdr kanye ne-cocgdz probe eklanywe ngokwesisekelo sazo yahlolwa kusetshenziswa uhlelo lwe-BLAST (www.ncbi.nlm.nih.gov/blast) kukho konke ukulandelana kofuzo lwe-glycerol triphosphate dehydrogenase yezinhlobo zohlobo lwe-Colletotrichum nezinye izinto eziphilayo ezitholakala kusizindalwazi se-GenBank.
Azikho izifunda ze-DNA zezinye izinto eziphilayo ezifana kakhulu neziqalo kanye nophenyo ezitholakele.
Ukuzwela kwesistimu yokuhlola kwahlolwa kusetshenziswa amasampula anezingxube ezihlukene ze-C. coccodes DNA, i-DNA evela eqabungeni lezambane elitheleleke nge-anthracnose (eqoqwe ngo-2017 e-Mari El, i-Red Scarlett ehlukahlukene), kanye nekhasi lezilimo eziyizigaxa ezithintekile indawo emnyama (eqoqwe esifundeni saseKostroma, izinhlobonhlobo ze-Red Scarlett, Ithebula 2). Ukuqinisekisa ukuba khona kwe-DNA ezigabeni nasezihlahleni zamazambane, izinhlobo ze-C. coccodes zahlukaniswa kuzo zaba amasiko ahlanzekile.
Imiphumela yokuhlaziywa kokuzwela kwesistimu yokuhlola ibonisa ukuthi ingasetshenziswa ukuxilonga ngempumelelo ubukhona be-C. coccodes DNA kusampula lapho okuqukethwe kwayo okuphelele kungxube ye-PCR kungaphezu kuka-0.05 ng. Lokhu kwanele ukubonwa, njengoba i-sclerotia eyodwa iqukethe, ngokwesilinganiso, i-0.131 ng, futhi inhlamvu eyodwa iqukethe cishe u-0.04 ng we-DNA (Cullen et al., 2002). Uhlelo lokuhlola olwakhiwe yiqembu lesiNgisi (Cullen et al., 2002) lubonise ukuzwela okufanayo (umjikelezo we-threshold 34 ku-0.05 ng DNA kanye ne-37 ku-0.005 ng).
Ukuhlaziywa kwamasampuli emvelo aqukethe amakhokhode e-C kuzo zonke izimo kwenze kwaba nokwenzeka ukudalula ngokuthembekile ubukhona bawo kusampula (Ithebula 2). Indlela ehlongozwayo yokuhlukaniswa kwe-DNA yayisebenza nasekuhlaziyeni amasampula ezitshalo zemvelo.
Ithebula 2. Ukunqunywa kokuzwela kwesistimu yokuhlola ehlongozwayo yokuhlonza amakhodi e-Colletotrichum ye-PCR yesikhathi sangempela.
Образец | Inani le-DNA kusampula *, ng | Umjikelezo weThreshold | C. ukutholwa kwamakhodi |
---|---|---|---|
I-Mycelium Colletotrichum coccodes | 50 | 21.3 | + |
5 | 25.7 | + | |
0.5 | 29,7 | + | |
0.05 | 33.5 | + | |
0.005 | 40 | - | |
0.0005 | 42.8 | - | |
0.00005 | - | ||
Ikhasi le-Tuber 1 | 50 | 32 | + |
Ikhasi le-Tuber 2 | 50 | 30 | + |
Ikhasi le-Tuber 3 | 50 | 31.5 | + |
Iqabunga lezambane | 50 | 29.5 | + |
Qaphela. * Engxubeni yemikhiqizo ye-PCR.
Ukuhlola ukucaciswa kwesistimu yokuhlola kwenziwa kumasampula e-DNA akhishwe ezinhlotsheni eziyi-15 zesikhunta. Zonke izinhlobo zesikhunta zahlukaniswa ngababhali bezithelo ezithintekile nezinempilo kanye namaqabunga katamatisi, izilimo eziyizigaxa zamazambane; uhlobo olulodwa lwahlukaniswa empandeni kakolweni (Ithebula 1). Phakathi kwalezo eziqhelile ebusweni besithelo, kukhona nezinhlobo ezingezona i-pathogenic kutamatisi (isibonelo, i-Phellinus ferrugineovelutinus).
Ucwaningo luye lwabonisa ukuthi i-C. coccodes DNA itholwe kumjikelezo we-threshold ka-20-27, kuyilapho ezinye izinhlobo zesikhunta azizange zibonwe noma zinikeze isignali ngemva komjikelezo we-40, okungabalelwa kumphumela womsindo ongaqondile (Ithebula 3).
Ithebula 3. Ukuhlola uhlelo lokuhlola izinhlobo ezahlukene zamakhowe
Igama lamakhowe | Umjikelezo weThreshold |
Amakhodi we-Colletotrichum 1 | 20.9 |
C. amakhodi 2 | 22.6 |
C. amakhodi 3 | 23 |
C. amakhodi 4 | 22 |
I-Fusarium oxysporum | > 40 |
F. verticalillium | > 40 |
I-Rhizoctonia solani | > 40 |
I-Phomopsis Phaseoli | > 40 |
I-Alternaria alternata | > 40 |
A. tomatophila | > 40 |
I-Helminthosporium solani | > 40 |
Phellinus ferrugineovelutinus | > 40 |
I-Stemphylium vesicarium | > 40 |
I-Ilyonectria crassa | > 40 |
I-Cladosporium cladosporioides | > 40 |
C. fulvum | > 40 |
I-Acrodontium luzulae | > 40 |
I-penicillium sp. | > 40 |
Qaphela. * Inani le-DNA kuwo wonke amasampula laliyi-10 ng.
Uhlelo lokuhlola oluthuthukisiwe lwasetshenziselwa ukukhomba ama-C. cocodes kumasampula amaqabunga katamatisi anezimpawu ze-necrotrophic pathogens nezilimo eziyizigaxa zamazambane ngaphandle kwezimpawu ezibonakalayo. Ocwaningweni, sithathe izilimo eziyizigaxa zembewu ezinhlobonhlobo ezitshalwe ezindaweni zaseKostroma, eMoscow, eKaluga, eNizhny Novgorod. Ukuba khona kwe-C. coccodes DNA kwakubhekwa njengokubalulekile kumasampuli, ekuhlaziyweni lapho umjikelezo we-threshold ungazange udlule u-35. Leli nani le-threshold likhethwe ngokusekelwe ekunqumeni okuthembekile kwe-0.05 ng ye-C. coccodes DNA (umjikelezo we-threshold 33.5, Ithebula 2) kanye neqiniso lokuthi imijikelezo ye-threshold engaphezu kuka-40, i-DNA engacacisiwe yezinye izinhlobo zesikhunta yatholwa. Ngale ndlela, ukutholakala okuphawulekayo kwe-C. coccodes DNA kutholwe kumasampuli angu-5 wezilimo eziyizigaxa ezitshalwe ezindaweni zaseKostroma, eMoscow, eKaluga naseqabungeni elilodwa likatamatisi elivela esifundeni sase-Yeisk esifundeni saseKrasnodar (Amathebula 4, 5).
Ithebula 4. Ukutholwa kwe-Colletotrichum coccodes ezigabeni zamazambane *
Inombolo yesampula | Amazambane ahlukahlukene | Indawo yokukhula | C. ukutholwa kwamakhodi | Umjikelezo weThreshold |
---|---|---|---|---|
1 | I-Red Scarlet | Isifunda saseKostroma | + | 35 |
2 | + | 35 | ||
3 | - | 38 | ||
4 | Sante | Isifunda saseMoscow | + | 34 |
5 | - | |||
6 | - | 41 | ||
7 | - | 41.8 | ||
8 | + | 30 | ||
9 | Zhukovsky ekuqaleni | Isifunda saseMoscow | - | 40.5 |
10 | - | 40.6 | ||
11 | - | |||
12 | UMolly | Isifunda saseKaluga | + | 34.3 |
13 | - | 38.4 | ||
14 | Amaphupho | Isifunda saseKaluga | - | |
15 | Gala | Isifunda saseNizhny Novgorod. | - | |
16 | - |
Qaphela. * Inani le-DNA kuwo wonke amasampula laliyi-50 ng.
Ithebula 5. Ukutholwa kwe-Colletotrichum coccodes emaqabunga katamatisi *
Inombolo yesampula | Indawo yokukhula | C. ukutholwa kwamakhodi | Umjikelezo weThreshold |
---|---|---|---|
1 | I-Krasnodar Territory, esifundeni saseCrimea | - | |
2 | - | ||
3 | - | ||
4 | - | 45 | |
5 | - | ||
6 | - | ||
7 | - | ||
8 | - | ||
9 | I-Krasnodar Territory, esifundeni sase-Yeisk | - | 39.2 |
10 | - | 40.8 | |
11 | - | ||
12 | - | 41.6 | |
13 | - | 40 | |
14 | - | 41 | |
15 | - | 41.9 | |
16 | - | ||
17 | - | ||
18 | - | 40.3 | |
19 | - | ||
20 | - | ||
21 | + | 34.5 | |
22 | - | ||
23 | - |
* Inani le-DNA kuwo wonke amasampula lalingama-50 ng.
Uhlelo lokuhlola oludalwe yithi alukho ngaphansi kwalolo oluthuthukiswe abacwaningi baseBrithani (uCullen et al., 2002) ngokuzwela nokucacisa futhi lufanele ukuhlaziya amasampula ezitshalo. Ukusetshenziswa kwayo kokuhlaziywa kwezilimo eziyizigaxa zembewu kwenza kwaba nokwenzeka ukuhlonza i-C. coccodes DNA kuma-tubers ngaphandle kwezimpawu zangaphandle zokulimala kanye nokuhlaziya ngempumelelo ukutheleleka kwamaqabunga.
Kuze kube manje, akukho ukuhlaziywa kwezilimo eziyizigaxa zamazambane zokuhlaselwa kwe-C. coccodes okwenziwe eRussia. Ucwaningo lwethu lokuqala lubonise ukuthi kwezingu-16 izilimo eziyizigaxa zembewu ezihloliwe ezitshalwe ezindaweni ezahlukene zeRussian Federation, ezi-5 ziqukethe ama-C. cocodes. Lokhu kukhombisa ukuthi indawo emnyama yezilimo eziyizigaxa zamazambane yisifo esijwayelekile samazambane eRussia, futhi indima yaso ekwehliseni umthamo kanye nekhwalithi yesitshalo samazambane ithathwa kancane.
Ukuhlaziywa kwamaqabunga katamatisi kwembula ukuba khona okuphawulekayo kwe-C. coccodes DNA eqabungeni elilodwa elivela esifundeni sase-Yeisk sase-Krasnodar Territory. Ngaphambilini, lapho kuhlolwa amasimu katamatisi eningizimu yeRussia kusetshenziswa uhlelo lokuhlola lwaseBrithani (Cullen et al., 2002), amaqabunga aqukethe ama-C. coccodes atholakala, futhi kwezinye izinkambu kwatholakala ingxenye enkulu yamaqabunga angenwe yi-C. coccodes (Belov et al., 2018). E-Krasnodar nasePrimorsky Territories, esifundeni saseMoscow, sithole izithelo zamatamatisi, lapho sakwazi khona ukuhlukanisa amasiko ahlanzekile e-C. coccodes. Kungenzeka ukuthi i-C. coccodes isakazeke kakhulu kutamatisi eRussia kunalokho okukholelwayo njengamanje, futhi ukulimaza kwayo nakho kubukelwa phansi.
Ngakho-ke, kuze kube manje, sekuqoqwe ukwaziswa okwanele ngokusakazwa kabanzi kwama- C. cocodes kumazambane notamatisi.
Ukuze uqonde kangcono indima yalesi sikhunta ekuthuthukiseni izifo zamazambane kanye notamatisi, kuyadingeka ukuqapha ukusabalala kwayo eRussia, ukutadisha indima yokutheleleka kwenhlabathi nembewu, kanye nendima yendawo emnyama ekulahlekelweni ngesikhathi sokugcinwa. Ukusetshenziswa kokuxilongwa kwe-PCR kungawenza lula lo msebenzi, futhi ukusetshenziswa kanyekanye kwazo zombili izinhlelo zokuhlola kuzokwandisa kakhulu ukunemba kokuhlaziya.
Lo msebenzi usekelwe isibonelelo esivela ku-Russian Science Foundation No. 18-76-00009.
Lesi sihloko sanyatheliswa kumagazini othi "Mycology and Phytopathology" (umqulu 54, No. 1, 2020).