Idya. Chudinov, V.A. Platonov, A.V. Alexandrova, S.N. Elansky
Munguva pfupi yapfuura, zvakaratidzwa kuti iyo ascomycete fungus Ilyonectria crassa inokwanisa kutapukira matapira tuber. Iri basa ndere rekutanga kuongorora hunhu hwehupenyu uye kushora kune mamwe mafungicides eiyo I. crassa strain yakaparadzaniswa nembatatisi. Iwo maratidziro emarudzi-akasarudzika matunhu eiyo "mbatatisi" kuenderana kwakabatana neiya akawanikwa pakutanga fungi yakaparadzaniswa kubva pamidzi yenarcissus, ginseng, aspen uye beech, maruva emaruva uye mashizha e tulip. Sezvineiwo, akawanda emusango uye gadheni zvirimwa zvinogona kuve matura eI. Crassa. Tsvagurudzo yakatsvagiswa yakatapukira madomasi netapatata, asi haina kutapudza muchero wemadomasi wese uye isina kuenzana mbatatisi tuber. Izvi zvinoratidza kuti I. crassa chirwere chevanga. Kuongororwa kwekurwisa fludioxonil, difenoconazole uye azoxystrobin pane yakasvibirira svikiro yakaratidza kushanda kwakanyanya kwemishonga iyi.
Index yeEC50 (iyo fungalion yeiyo fungicide, iyo inononoka kudzika nekaviri nguva yekukura kwakakura kwenzvimbo inoenderana neiyo isiri fungicide kudzora) yaive yakaenzana ne2; 0.4 uye 7.4 mg / l, zvichiteerana. Iko mukana wekuvandudzwa kwechirwere chakakonzerwa naI.crassa inofanirwa kutariswa kana phytopathological kuongororwa kwemazamu tubers uye nekuvandudzwa kwenzvimbo dzekuchengetedza dura.
Iko kukura kwe phytopathogenic microorganisms kunotungamira kukurasikirwa kwakanyanya pamatanho ese ekurima nekuchengetedza mbatatisi. Paunenge uchironga matanho ekudzivirira, sekutonga, anozivikanwa marwisi anofungidzirwa, senge marudzi egenera Alternaria, Fusarium, Phoma, Helminthosporium, Colletotrichum, Phytophthora, nezvimwewo. Zvisinei, mumakore achangopfuura, mimwe mishumo yakaonekwa. pamusoro pekuonekwa kweatsva phytopathogenic microorganisms pamatatata. Biology yavo haina kunyatso dzidzwa, kushanda kwemafungicides anoshandiswa pamatatata maererano navo hazvizivikanwe, nzira dzekuongorora hadzina kugadzirwa. Nekukura kwakawanda, ivo vanokwanisa kukonzeresa kukuvara kukuru kune chirimwa chembatatisi. Imwe yema microorganisms aya ndeayo ascomycete fungus Ilyonectria crassa (Wollenw.) A. Cabral & Crous, yakatanga kuwanikwa nevanyori vembatatari tubers (Chudinova et al., 2019).
Iri basa rinopa mhedzisiro yekuongororwa kweiyo I. crassa strain yakasarudzika kubva kune iyo mbatatari tuber. Iyo morphology yemakoloni uye mycelial zvivakwa zveI. Crassa, nucleotide kuteedzana kwemarudzi-akasarudzika DNA matunhu, huturu kumatato nematomatisi, uye kuramba kune mamwe anozivikanwa mafungicides akaongororwa.
Zvishandiso uye nzira
Isu takashandisa iyo I. crassa 18KSuPT2 strain yakasarudzika muna 2018 kubva kune yakakanganiswa mbatatari tuber yakakura munzvimbo yeKostroma. Iyo tuber yakakanganiswa neyakaoma kuora nekavha yakafukidzwa neyakajeka brown brown mycelium. Ichishandisa tsono yekuparadzanisa isina kubereka, iyo fungal mycelium yakaendeswa mune yePetri dhishi ine agar medium (doro wort 10%, agar 1.5%, penicillin 1000 U / ml). Mahwendefa akaiswa murima pa24 ° C.
Leica DM2500 light microscope ine ICC50 HD digital camera uye Leica M80 binocular microscope ine IC80HD digital camera (Leica Microsystems, Germany) yakashandiswa kutora mifananidzo, kuongorora saizi uye morphology yezvipore uye nhengo dzepore.
Kupatsanura DNA, iyo fungal mycelium yakasimwa mumvura pea svikiro, ndokuzadzwa nechando muine nitrogen yemvura, homogenized, yakavharirwa muCTAB buffer, yakanatswa ne chloroform, uye yakashambidzwa kaviri ne 2% doro.
Iyo nzira yekubvisa yeDNA inotsanangurwa zvakadzama muchinyorwa naKutuzova et al. (2017).
Kuona marudzi nema molecular nzira uye kuenzanisa nemamwe anozivikanwa I. crassa strains, PCR yakaitwa pamwe nemitangaridzo ichibvumira kukwidziridzwa kwenzvimbo dzakasiyana-siyana dzeDNA matunhu: ITS1-5,8S-ITS2 (primers ITS5 / ITS4, White et al., 1990), gene matunhu b -tubulin (Bt2a / Bt2b, Girazi, Donaldson, 1995) uye dudziro elongation factor 1cy (tef1cy) (primers EF1-728F / EF1-986R, Carbone naKohn, 1999). Amplicons ehurefu hwaidiwa aibviswa kubva mujel achishandisa Evrogen CleanUp kit. Matunhu akawedzeredzwa akateedzana achishandisa BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, CA, USA) pane Applied Biosystems 3730 xl otomatiki sequencer (Applied Biosystems, CA, USA). Mhedzisiro yeiyi nucleotide yakateedzana yakashandiswa kutsvaga mutambo muDenBank dhatabhesi yeUS National Center yeBiotechnology Ruzivo (NCBI). Phylogenetic ongororo yakaitwa uchishandisa iyo MEGA 6 chirongwa (Tamura et al., 2013).
Kutsunga kwehutachiona kwakaitwa pazvibereko zvakasvibira zvemadomasi makuru-akachekwa (akasiyana Dubrava) uye mbatatari tubers (dzakasiyana Gala). Uye zvakare, zvidimbu zvemichero imwechete uye machubhu aishandiswa kuteedzera kukuvara kwemichero yakakanganiswa nemachubhu. Zvidimbu zvematubhu zvakaiswa mumakamuri ane hunyoro, ayo aive Petri madhishi ane pepa nyoro repasi pazasi. Slide yakaiswa papepa, iro, zvakare, zvidimbu zvematubhi kana michero yakaiswa. Yese tubers nemichero zvakaiswawo mumidziyo ine nyoro nyoro bepa pazasi. Pakati pechidimbu (kana panzvimbo isina kugadzikana yeiyo tuber kana michero), chidimbu cheagar (5 × 5 mm) ine fungal hyphae yakaiswa mushure memazuva mashanu ekukura pa wort agar.
Kuongororwa kwekushomeka kwefungus tambo kune fungicides kwakaitwa mumarabhoritari mamiriro pane agar zvinovaka muviri svikiro. Takadzidza kugona kubata fungicidal zvinodhaka Maxim, KS (chinoshanda chirungiso fludioxonil, 25 g / l), Quadris, KS (azoxystrobin 250 g / l), Scor, EC (difenoconazole 250 g / l) (State catalog ..., 2020 ). Kuongorora kwakaitwa muPetri ndiro pane wort-agar svikiro pamwe nekuwedzeredzwa kwemishonga yakadzidzwa pakumisikidzwa kwechinhu chinoshanda che0.1; Poshi; 1 ppm (mg / L) (ye fludioxonil uye difenoconazole), 10; gumi; 1 ppm (yeazoxystrobin) uye mune midhiya isina fungicide (kutonga). Iyo fungicide yakawedzerwa kune yakanyungudutswa svikiro yakatonhora kusvika 10 ° C, mushure mezvo svikiro rakadururwa muPetri ndiro. Agar block ine fungal mycelium yakaiswa pakati pePetri dhishi uye yakarongedzwa patembiricha ye100 ° C murima. Mushure memazuva manomwe ekupukuta, madhaimendi emakoloni akayerwa munzira mbiri dzekutenderera; zviyero zveyero yekoloni yega yega zvakaenzana. Kuedza kwakaitwa katatu. Kubva pamhedzisiro yeongororo, iyo EC60 yakaverengerwa, yakaenzana neyakaunganidzwa yefungicide, iyo yakadzikisa chiyero chekukura kwakakura kwenzvimbo inoenderana neiyo fungicide-yemahara kutonga.
Mhedzisiro uye hurukuro
Pane petri ndiro ine wort agar, iyo fungus yakaumba makoloni ane chena flocculent mycelium. Svikiro pasi peiyo mycelium rakashanduka rakatsvukuruka-shava. Kana iyo svikiro yaoma, iyo fungus yakaumba mhodzi dzemhando mbiri pane imwechete uye yakaunganidzwa conidiophores mune diki sporodochia. Macroconidia akareba, cylindrical, aine imwechete kusvika matatu septa, avhareji kureba 27.2 µm aine huwandu hwetsika kubva ku23.2 kusvika 32.2 µm, upamhi kusvika 4.9 µm (Fig. 1). Ivhareji kureba kwemicroconidia iri 14.3 µm ine huwandu hwehunhu kubva pa10.3 kusvika 18.1 µm, uye upamhi hwasvika ku4.0 µm. Ese macro- uye micromorphological vatambi vanokodzera munzvimbo dzakasiyana dzekusiyana kwemarudzi Ilyonectria crassa (Cabral et al., 2012).
Iwo maratidziro enzvimbo-akasarudzika DNA matunhu (ITS, b-tubulin, TEF 1cy) zvakanyatsoenderana nekuteedzana kweiyo I. crassa strains dzatakadzidza kare (Chudinova et al., 2019, Tafura 1). Kuti ndiongorore kuwanda kweI.crassa mune mamwe matunhu uye kuongorora iwo maratidziro etsika dzakakanganiswa, zvakafanana DNA zviyero muGenBank dhatabhesi zvakaongororwa (Tafura 1). Kubatana kwacho kwaive 86 kusvika 100%. Kurongeka kwenzvimbo dzese nhatu dzeDNA dze "mbatatisi" I. crassa strain yaive yakafanana nekuteedzana kwematambudziko akasiyiwa kubva kune lily girobhu uye daffodil midzi muNetherlands uye kubva kune ginseng midzi muCanada. Hatina kukwanisa kuwana mamwe I. crassa strains ane matatu akaongororwa anoteedzana akaenzana mune akavhurika dhatabhesi. Nekudaro, kuongororwa kweakachengeterwa ITS uye b-tubulin akateedzana zvakaratidza kuvepo kwaI. Crassa pamashizha e tulip muUK. Fungi ine akafanana ITS akateedzana akaonekwa pakuongorora mycobiota yeaspen midzi muCanada uye beech midzi muItari, uye mbatatari tubers muSaudi Arabia (Tafura 1). Mhedzisiro yetsvagiridzo iyi inoratidza kuti ini. Crassa ine kugoverwa kwepasirese uye inokwanisa kutapudza marudzi akasiyana emiti.
Pakusarudza hutachiona pazvidimbu zvemadomasi uye mbatatisi pazuva rechishanu, hupamhi hweronda hwasvika 5 cm.Muchiitiko ichi, iyo yakatsvagiswa strain haina kutapudza iyo yese yemadomasi michero uye isina kujeka mbatatisi tuber. Zvisinei, sepals dzakakanganiswa padomasi. Kuti ubvise mukana wekusvibiswa, fungal isolate kubva ku mycelium yakagadziriswa pane mbatatisi tuber slice yakasarudzika mutsika yakachena. Izvo zvaive zvakanyatso nyatso kufanana neshungu yemubereki. Sezvineiwo, ini. Crassa chirwere chevanga.
Kugara-kudyara kurapwa kwembeu tubers ne fungicides kunoderedza kukura kwezvirwere pamiti panguva yekukura mwaka. Nezvekusarudzwa kweanoshanda fungicides, zvakakosha kuti uongorore kuti ndedzipi dzadzo dzinoshanda pakurwisa I. сrassa. Basa rakadzidza zvakapararira zvinoshanda zvinhu zvefungicides - fludioxonil, azoxystrobin, difenoconazole. Fludioxonil inogadzirwa mumisanganiswa yakati wandei inoshandiswa pakupfekedza mbeu nembeu tubers usati wadyara. Fludioxonil (Maxim) inoshandiswa zvakare kurapa mhodzi yemidziyo isati yachengeterwa. Difenoconazole uye azoxystrobin inosanganisirwa mune dzinoverengeka gadziriro dzinoshandiswa kugadzirisa zvinhu zvembeu, pamwe nekugadzirira kunoitirwa kugadzirisa zvinomera (State catalog ..., 2020).
Kukura kweiyo I. crassa yaidzidziswa pane midhiya (Fig. 2) ine akasiyana masisitimu ezvinhu zvinoshanda: fludioxonil (EC50 = 0.4 ppm), azoxystrobin (EC50 = 4 ppm), uye difenoconazole (EC50 = 7.4 ppm) (Tafura 2 ). Aya magadzirirwo anogona kutariswa seanoshanda kwazvo kurwisa I. crassa, sezvo yavo EC50 iri yakaderera zvakanyanya kupfuura iyo yakakurudzirwa kukosheswa kwekugadzirira mune inoshanda mvura inoshandiswa pakurapa tubers. Zvinoenderana neHurumende Catalog ... (2020), kuwanda kwefludioxonil iri mumvura yekurapa mbatatari tubers kunobva pa500 kusvika 1000 ppm, azoxystrobin (iri mumvura yekurapa pasi pehororo) - 3750-9375 ppm, difenoconazole (mune yemvura yekurapa zvinomera zvinomera) - 187.5-625 ppm.
Tafura 1. Kufanana kwekuteedzana kwemarudzi-akasarudzika maseru e18KSuPT2 uye maronda eIlyonectria crassa anowanikwa mudura reGenbank
Kushungurudzika | Imiti yekuchengetedza, nzvimbo yekubuda | Sequence nhamba dzakaiswa muGenBank, muzana yezvakafanana | batanidzo | ||
STIs | β-tubulin | TEF 1cy | |||
17KSPT1 uye 18KSuPT2 | Mbatata tuber, nzvimbo yeKostroma | MH818326 | MH822872 | MK281307 | Chudinova et al., 2019, iri basa |
CBS 158/31 | Narcissus midzi, Netherlands | JF735276 100 | JF735394 100 | JF735724 99.3 | Cabral et al., 2012 |
CBS 139/30 | Lily girobhu racho, Netherlands | JF735275 100 | JF735393 99.7 | JF735723 99.3 |
|
NSAC-SH-1 | Ginseng mudzi, Canada | AY295311 99.4 | JF735395 100 | JF735 / 725 99.6 |
|
RHS235138 | Tulip shizha, UK | KJ475469 100 | KJ513266 100 | ND | Denton, Denton, 2014 |
MT294410 | Aspen midzi, Canada | MT294410 100 | ND | ND | Ramsfield et al., 2020 |
ER1937 | Beech, Itari | KR019363 99.65 | ND | ND | Tizzani, Haegi, Motta. Kutumira kwakananga |
KAUF19 | Mbatata tuber, Saudi Arabia | HE649390 98.3 | ND | ND | Gashgari, Gherbawy, 2013 |
ND = isina kuiswa
Tafura 2. Kuramba kweIlyonectria crassa kune fungicides
(chinhu chinoshanda) | EC50, ppm | ||||
3 zuva | 5 zuva | 7 zuva | |||
Kudzora | 17 ± 2 | 33 ± 5 | 47 ± 3 | ||
Quadris, KS (fsoxystrobin) | 18 ± 1 | 34 ± 2 | 48 ± 2 | ||
11 ± 1 | 11 ± 1 | 12 ± 1 | |||
11 ± 1 | 11 ± 1 | 12 ± 1 | |||
Kuwanda, KS (fludioxonil) | 16 ± 1 | 28 ± 2 | 48 ± 2 | ||
7 ± 1 | 13 ± 3 | 19 ± 4 | |||
5 ± 1 | 12 ± 1 | 17 ± 5 | |||
Skor, EC (difenoconazole) | 18 ± 1 | 35 ± 2 | 48 ± 1 | ||
11 ± 1 | 24 ± 3 | 35 ± 4 | |||
11 ± 1 | 13 ± 1 | 17 ± 3 |
Mubasa redu, I. crassa strains dzaive dzakasarudzika kubva ku mbatatari tubers munzvimbo dzeKostroma neMoscow (Chudinova et al., 2019) matunhu. Chikamu chakakwira chefungal strains ine ayo akateedzana akafanana naI. Crassa yakaratidzwa mukuwongorora mycobiota yematato tubers muSaudi Arabia (Gashgari naGherbawy, 2013). Sezvineiwo, ini. Crassa haisi isingawanzo pamatezi sezvazvingaite. Mukuyedza kwedu, zvakaratidzwa kuti fungus inogona kukanganisa michero yemadomasi. Inozivikanwa kubva muzvinyorwa izvo I. crassa inokwanisa kusimudzira saprotrophically muvhu (Moll et al., 2016), pamwe nekukanganisa zvakasiyana siyana zvezvirimwa, kunyangwe zvimiro zvemitero zviri kure zvakadai sedaffodils, maruva, ginseng, aspen, uye beech (Tafura 1). imwe). Sezvineiwo, akawanda emusango uye gadheni zvirimwa zvinogona kuve matura eI. Crassa. Izvo zviri pamusoro zvinoratidza kuti kana uchigadzira matanho ekudzivirira, zvakafanira kufunga nezve mukana wekukanganisa matapiri emazamu nefungus iyi. Kugadzirira kwakapararira kwekurapa mbatatari tubers ine fludioxonil, azoxystrobin uye difenoconazole yakaratidza kushanda kwakanyanya kwefungicidal kurwisa I. crassa.
Iri basa rakatsigirwa neRussia Foundation Yekutanga Yekutsvagisa (Grant Nha. 20-016-00139).
Chinyorwa ichi chakaburitswa mumagazini "Dziviriro Dziviriro", 2020, 103 (3)