NOJ. Chudinov, V.A. Platonov, A.V. Alexandrova, S.N. Elansky
Nws tau tsis ntev los no tau pom tias ascomycete fungus Ilyonectria crassa muaj peev xwm kis cov qos ntoo. Cov haujlwm no yog thawj zaug los tshawb xyuas qhov txheeb raws roj ntsha thiab tiv taus qee cov fungicides ntawm I. crassa lim cais los ntawm cov qos yaj ywm. Kev sib txawv ntawm hom tsiaj thaj av ntawm "qos" kab sib xyaw nrog cov uas tau ua ntej rau cov kab mob fungi cais tawm ntawm cov hauv paus ntawm daffodil, ginseng, aspen thiab beech, nplooj roj thiab tulip nplooj. Thaj, ntau hom tsiaj qus thiab vaj tuaj yeem yog kev pom zoo ntawm I. crassa. Kev soj ntsuam kab mob kis tau cov txiv lws suav thiab qos yaj ywm, tab sis tsis kis rau tag nrho cov txiv lws suav thiab hauv cov qos ntoo. Qhov no qhia tau tias I. crassa yog kev mob cab. Kev soj ntsuam ntawm kev tsis kam rau fludioxonil, difenoconazole thiab azoxystrobin ntawm cov txiv ntseej nruab nrab pom tau hais tias qhov ua tau zoo ntawm cov tshuaj no.
Qhov ntsuas qhov ntsuas EC50 (kev tswj hwm ntawm fungicide, uas ua kom qis qis dua 2 zaug ntawm tus nqi ntawm radial kev loj hlob ntawm qhov sib piv rau cov tsis muaj tshuaj tiv thaiv fungicidal) tau sib npaug rau 0.4; 7.4 thiab 4 mg / l, ntsig txog. Muaj peev xwm ntawm kev txhim kho ntawm tus kab mob tshwm sim los ntawm I. crassa yuav tsum tau coj mus rau hauv tus account thaum phytopathological kev ntsuas ntawm cov qos yaj ywm thiab kev txhim kho ntawm cov nroj tsuag tiv thaiv kev ntsuas.
Kev tsim cov phytopathogenic microorganisms ua rau muaj kev poob zoo nyob rau txhua theem ntawm kev loj hlob thiab khaws cia qos yaj ywm. Thaum npaj kev tiv thaiv kev tiv thaiv, raws li txoj cai, cov kab mob uas paub zoo tau coj mus rau hauv tus account, xws li hom kab mob ntawm genera Alternaria, Fusarium, Phoma, Helminthosporium, Colletotrichum, Phytophthora, thiab lwm yam. Txawm li cas los xij, hauv xyoo tsis ntev los no, ntau thiab ntau daim ntawv qhia tau tshwm sim ntawm cov tsos ntawm cov kab mob tshiab phytopathogenic rau ntawm cov qos yaj ywm. Lawv cov biology kawm tsis zoo, qhov ua tau zoo ntawm fungicides siv rau cov qos yaj ywm hauv kev sib raug zoo rau lawv tsis paub, cov txheej txheem kuaj mob tseem tsis tau tsim. Nrog rau kev txhim kho huab hwm coj, lawv muaj peev xwm ua rau muaj kev puas tsuaj rau cov qoob loo qos. Ib qho ntawm cov kab mob me me yog cov kab mob ascomycete fungus Ilyonectria crassa (Wollenw.) A. Cabral & Crous, thawj zaug tau tshawb pom los ntawm cov kws sau txog cov qos qos (Chudinova li al., 2019).
Cov haujlwm no nthuav tawm cov txiaj ntsig ntawm kev tsom xam ntawm I. crassa lim cais los ntawm cov qos qos. Cov qauv ntawm cov kob ntawm cov kab xev thiab cov qauv mycelial ntawm I. crassa, nucleotide ntu ntawm hom DNA thaj chaw, sib deev rau qos yaj ywm thiab txiv lws suav, thiab kev tawm tsam rau qee cov fungicides nrov tau kawm.
Cov ntaub ntawv thiab cov hau kev
Peb tau siv cov I. crassa 18KSuPT2 hom cais tawm hauv 2018 los ntawm cov tuber kis mob kis nyob hauv Kostroma cheeb tsam. Lub tuber tau cuam tshuam los ntawm hom qhuav yam nrog kab noj hniav uas npog nrog lub teeb xim av mycelium. Siv cov tshuaj tua kab mob uas tsis pom kev tawm, fungal mycelium tau pauv mus rau hauv Petri phaj nrog qhov nruab nrab agar (npias wort 10%, agar 1.5%, penicillin 1000 U / ml). Cov phiaj xwm tau muab tso rau qhov tsaus ntuj ntawm 24 ° C.
Lub tshuab tsom lub teeb pom kev zoo Leica DM2500 nrog ICC50 HD lub koob yees duab thiab lub koob yees duab Leica M80 nrog lub IC80HD digital lub koob yees duab (Leica Microsystems, Lub Tebchaws Yelemees) tau siv los thaij duab, ntsuas qhov loj me thiab morphology ntawm cov noob kab ntsig thiab spore kabmob.
Txhawm rau cais DNA, fungal mycelium tau zus rau hauv cov kua pea nruab nrab, tom qab ntawd khov rau hauv cov kua nitrogen, homogenized, muab tso rau hauv CTAB tsis, ntxuav nrog chloroform, thiab ntxuav ob zaug nrog 2% cawv.
Cov txheej txheem DNA rho tawm tau piav qhia txhua yam hauv tsab xov xwm los ntawm Kutuzova li al. (2017).
Txhawm rau txiav txim siab seb hom tsiaj los ntawm hom kev siv molecular thiab sib piv nrog lwm cov paub I. kab crassa, PCR tau nqa tawm nrog primers tso cai ua kom nrov ntawm cov hom DNA tshwj xeeb: ITS1-5,8S-ITS2 (primers ITS5 / ITS4, Dawb li al., 1990), cov cheeb tsam b. -tubulin (Bt2a / Bt2b, Iav, Donaldson, 1995) thiab kev txhais cov ntsiab lus elongation 1α (tef1α) (primers EF1-728F / EF1-986R, Carbone thiab Kohn, 1999). Cov kab mob ntawm qhov ntev yam xav tau raug muab rho tawm los ntawm cov gel uas siv cov khoom siv Evrogen CleanUp. Cov cheeb tsam muaj zog tau ua ntu zus siv BigDye® Terminator v3.1 Cov Cuab Yeej Siv Nruab Nrab (Siv Khoom Siv Hluav Taws Xob, CA, Tebchaws USA) ntawm Kev Siv Biosystems 3730 xl automated sequencer (Applied Biosystems, CA, USA). Cov txiaj ntsig nucleotide tau siv los tshawb ib qho kev sib tw hauv Asmeskas National Center for Biotechnology Information (NCBI) GenBank database. Kev soj ntsuam phylogenetic tau siv MEGA 6 qhov kev pab cuam (Tamura li al., 2013).
Kev txiav txim siab ntawm virulence tau nqa tawm ntawm cov txiv ntoo ntsuab tag nrho ntawm cov txiv lws suav loj (Dubrava ntau yam) thiab cov qos yaj ywm (Gala ntau yam). Tsis tas li ntawd, txhawm rau sim txiv rau cov txiv hmab txiv ntoo thiab cov tub hluas uas raug puas tsuaj, peb siv cov nplais ntoo ntawm tib cov txiv ntoo thiab cov tub. Cov hlais cov nqaj tau muab tso rau hauv cov chav noo, uas yog Petri cov tais diav nrog cov ntawv lim ntub rau hauv qab. Ib qho swb tau muab tso rau ntawm daim ntawv, uas, nyob rau hauv lem, slices ntawm tubers los yog txiv hmab txiv ntoo raug tso. Tag nrho lub hau thiab cov txiv ntoo kuj tau muab tso rau hauv cov thawv uas muaj cov ntawv lim ntub rau hauv qab. Nyob rau hauv qhov chaw ntawm cov hlais (lossis nyob rau ntawm qhov chaw mos ntawm lub tuber lossis txiv hmab txiv ntoo), ib daig ntawm agar (5 mm 5 hli) nrog fungal hyphae tau muab tso tom qab 5 hnub loj hlob ntawm wort agar.
Kev ntsuas qhov tsis kam ntawm kev tawm tsam ntawm cov kab mob rau cov kab mob fungicides tau ua nyob hauv chav kuaj mob ntawm agar cov khoom noj nruab nrab. Peb kawm txog qhov ua rau muaj kev raug mob rau cov tshuaj fungicidal Maxim, KS (nquag ua tshuaj fludioxonil, 25 g / l), Quadris, KS (azoxystrobin 250 g / l), Scor, EC (difenoconazole 250 g / l) (Xeev phau ntawv ..., 2020). Kev soj ntsuam tau nqa tawm hauv Petri cov tais diav ntawm wort-agar nruab nrab nrog ntxiv ntawm cov tshuaj tshawb fawb ntawm qhov ntau ntawm cov tshuaj nquag 0.1; ib tug; 1 ppm (mg / L) (rau fludioxonil thiab difenoconazole), 10; kaum; 1 ppm (rau azoxystrobin) thiab hauv xov xwm yam tsis muaj fungicide (tswj). Lub tshuab ua kom khov tau ntxiv rau yaj thiab txias kom txog 10 ° C nruab nrab, tom qab ntawd qhov nruab nrab tau muab hliv rau hauv Petri cov tais diav. Lub pob zeb hauv av nrog cov pwm mycelium tau tso rau hauv qhov chaw ntawm Petri phaj thiab kab lis kev cai ntawm qhov kub ntawm 100 ° C hauv maub. Tom qab 60 hnub ntawm kev tsim tawm, cov diameters ntawm cov pawg tau los ntawm kev ntsuas ua ob qho kev sib tshuam sib luag; cov kev ntsuas ntsuas rau txhua qhov ntev tau nruab nrab. Cov kev sim tau ua hauv triplicate. Raws li cov txiaj ntsig tau los ntawm kev tsom xam, EC24 tau suav, sib npaug ntawm cov tshuaj tua kab ntawm fungicide, uas tau txiav txim siab ib ntu ntawm radial kev loj hlob ntawm lub colony txheeb ze rau cov tswj fungicidal.
Cov txiaj ntsig thiab kev sib tham
On petri lauj kaub tais diav nrog wort agar, lub fungus tsim pawg nrog dawb flocculent mycelium. Qhov nruab nrab hauv qab mycelium tig liab-xim av. Thaum nruab nrab dries nce, lub fungus tsim spores ntawm ob hom rau ib leeg thiab zuag qhia conidiophores hauv sporodochia me me. Macroconidia yog elongated, cylindrical, nrog ib mus rau peb septa, qhov nruab nrab ntev 27.2 µm nrog qhov ntau ntawm qhov tseem ceeb ntawm 23.2 txog 32.2 µm, qhov dav - txog 4.9 µm (Daim duab 1). Qhov nruab nrab ntev ntawm microconidia yog 14.3 µm nrog qhov ntau ntawm qhov tseem ceeb ntawm 10.3 txog 18.1 µm, qhov dav dav mus txog 4.0 µm. Txhua hom macro- thiab cov cim micromorphological haum rau ntau hom kev hloov pauv ntawm hom Ilyonectria crassa (Cabral li al., 2012).
Cov kab ke ntawm cov cheeb tsam ntawm DNA tshwj xeeb (ITS, b-tubulin, TEF 1α) sib luag nrog cov kab sib txuas ntawm I. crassa hom peb tau kawm ua ntej (Chudinova li al., 2019, Rooj 1). Txhawm rau kawm txog kev nthuav dav ntawm I. crassa hauv lwm thaj chaw thiab tshuaj xyuas qhov pom ntawm cov kev coj noj coj ua, cov DNA zoo sib xws hauv GenBank database tau txheeb xyuas (Cov Lus 1). Kev sib tshooj yog 86 rau 100%. Cov xwm txheej ntawm tag nrho peb cheeb tsam DNA ntawm lub "qos" I. crassa hom tau zoo ib yam rau cov kab ntawm cov leeg mob cais tawm los ntawm cov ceg teeb thiab narcissus keeb kwm hauv Netherlands thiab los ntawm cov hauv paus ginseng hauv Canada. Peb tsis tuaj yeem nrhiav lwm tus I. crassa lim nrog peb peb tshuaj ntsuam qhov sib piv hauv cov ntsuas qhib. Txawm li cas los xij, kev soj ntsuam ntawm qhov tso nyiaj ITS thiab b-tubulin ua ntu zus pom tias muaj I. crassa ntawm tulip nplooj hauv tebchaws Askiv. Fungi nrog cov kab ke ntawm ITS zoo sib xws tau txheeb xyuas hauv kev soj ntsuam mycobiota ntawm aspen keeb kwm hauv Canada thiab beech keeb kwm hauv Ltalis, qos yaj ywm hauv Saudi Arabia (Cov Lus 1). Cov txiaj ntsig ntawm txoj kev kawm no qhia tau tias I. crassa muaj cov khoom xa tawm thoob ntiaj teb thiab muaj peev xwm kis ntau hom tsiaj.
Thaum txiav txim qhov pathogenicity ntawm cov nplais txiv lws suav thiab qos yaj ywm rau hnub 5, txoj kab uas hla ntawm lub qhov txhab ncav cuag 1.5 cm. Kev soj ntsuam kab mob tsis kis tau rau tag nrho cov txiv lws suav txiv ntoo thiab cov qos tuber. Txawm li cas los xij, cov sepals tau cuam tshuam rau ntawm cov txiv lws suav. Yuav kom cais tawm qhov ua tau ntawm kev sib kis los ntawm mycelium tsim nyob rau qos qos tuber hlais, ib qho fungal cais tawm mus rau kev coj noj coj ua dawb huv. Nws yog kiag li zoo li tus niam hom. Thaj, I. crassa yog kev mob cab.
Kev npaj ua ntej cog kev kho cov noob raj nrog fungicides txo qhov kev txhim kho ntawm cov kab mob ntawm cov nroj tsuag thaum lub caij cog qoob loo. Txog kev xaiv cov tshuaj fungicides zoo, nws yog ib qho tseem ceeb uas yuav tau ntsuam xyuas seb yam twg thiaj li ua tau zoo tiv thaiv I. сrassa. Kev ua haujlwm tau kawm txog kev nthuav dav cov tshuaj lom neeg muaj nyob rau hauv fungicides - fludioxonil, azoxystrobin, difenoconazole. Fludioxonil tau tsim nyob hauv ntau cov khoom sib xyaw siv rau kev hnav khaub ncaws noob thiab cov noob noob ua ntej cog. Fludioxonil (Maxim) kuj tseem siv tshuaj tua cov noob ua ntej cia khoom. Difenoconazole thiab azoxystrobin tseem muaj nyob rau hauv ntau qhov kev npaj siv rau kev ua tiav cov khoom noob, zoo li hauv kev npaj npaj rau kev ua cov nroj tsuag cog (Lub xeev cov ntawv sau ..., 2020).
Kev loj hlob ntawm I. crassa tau kawm hauv xov xwm (Daim duab 2) nrog ntau qhov sib txawv ntawm cov khoom xyaw nquag: fludioxonil (EC50 = 0.4 ppm), azoxystrobin (EC50 = 4 ppm), thiab difenoconazole (EC50 = 7.4 ppm) (Cov Lus 2). Cov kev npaj no tuaj yeem pom tias muaj txiaj ntsig zoo tiv thaiv I. crassa, txij li lawv qhov EC50 yog qhov qis dua qhov kev pom zoo ntawm kev npaj rau hauv cov kua dej ua haujlwm siv rau kev kho mob ntawm tub. Raws li lub Xeev Kev Qhia Ntawv ... (2020), qhov muaj siab ntawm fludioxonil hauv cov kua rau kev kho cov kab mob qos yog los ntawm 500 txog 1000 ppm, azoxystrobin (hauv cov kua rau kev kho hauv qab ntawm lub furrow) - 3750-9375 ppm, difenoconazole (hauv cov kua rau kev kho cov nroj tsuag tsis zoo) - 187.5– 625 ppm.
Cov lus 1. Seem sib xws ntawm cov tsiaj-raws qib raws dua ntawm hom 18KSuPT2 thiab hom ntawm Ilyonectria crassa muaj nyob hauv Genbank database
Lub strain | Tus tswv tsev cog, hauv tsev kawm ntawv ntawm kev nthuav dav | Cov lej naj npawb tau muab tso rau hauv GenBank, feem pua ntawm qhov sib xws | txuas | ||
ITS | β-tubulin | TEF 1 α | |||
17KSPT1 thiab 18KSuPT2 | Qos tuber tuber, cheeb tsam Kostroma | MH818326 | MH822872 | MK281307 | Chudinova li al., 2019, qhov haujlwm no |
SIB 158/31 | Cov hauv paus loj ntawm Narcissus, Tebchaws Netherlands | JF735276 100 | JF735394 100 | JF735724 99.3 | Cabral li al., 2012 |
SIB 139/30 | Lily cov noob, Netherlands | JF735275 100 | JF735393 99.7 | JF735723 99.3 |
|
NSAC-SH-1 | Ginseng paus, Canada | AY295311 99.4 | JF735395 100 | JF735 / 725 99.6 |
|
RHS 235138 | Tulip nplooj, UK | KJ475469 100 | KJ513266 100 | ND | Denton, Denton, xyoo 2014 |
MT294410 | Aspen keeb kwm, Canada | MT294410 100 | ND | ND | Ramsfield li al., 2020 |
ER1937 | Beech, Ltalis | KWB 019363 99.65 | ND | ND | Tizzani, Haegi, Motta. Kev xa tawm ncaj qha |
QAB 19 | Qos yaj ywm tuber, Saudi Arabia | HE649390 98.3 | ND | ND | Gashgari, Gherbawy, 2013 |
ND = tsis tso nyiaj
Cov lus 2. Tsis kam ntawm Ilyonectria crassa rau fungicides
(tseem muaj khoom) | EC50, ppm | ||||
3 hnub | 5 hnub | 7 hnub | |||
Kev tswj | 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 | |||
Maxim, 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 |
Hauv peb txoj haujlwm, I. crassa hom raug cais los ntawm cov qos yaj ywm hauv Kostroma thiab Moscow (Chudinova li al., 2019) thaj chaw. Qhov feem ntau ntawm cov kab mob fungal nrog ITS cov kab sib luag zoo ib yam li I. crassa tau qhia tawm thaum txheeb xyuas mycobiota ntawm cov qos yaj ywm hauv Saudi Arabia (Gashgari and Gherbawy, 2013). Thaj, I. crassa tsis tshua tsawg ntawm qos yaj ywm li nws yuav zoo li. Peb cov kev sim pom tias cov pwm no tuaj yeem cuam tshuam cov txiv lws suav txiv ntoo. Nws paub los ntawm cov ntaub ntawv tias I. crassa muaj peev xwm txhim kho saprotrophically hauv av (Moll et al., 2016), thiab kis tau rau ntau hom nroj tsuag, txawm tias taxonomically nyob deb suav daws xws li daffodils, lilies, ginseng, aspen, thiab beech (Cov Lus 1). ib). Thaj, ntau hom tsiaj qus thiab vaj tuaj yeem yog kev pom zoo ntawm I. crassa. Qhov saum toj no qhia tau hais tias thaum tsim kev tiv thaiv kev tiv thaiv, nws yog ib qho tsim nyog yuav tsum coj mus rau hauv tus account qhov tseem cuam tshuam ntawm cov qos yaj ywm nrog cov kab mob fungus no. Kev npaj dav dav rau kev kho cov qos yaj ywm muaj cov fludioxonil, azoxystrobin thiab difenoconazole tau pom tias muaj qhov ua tau zoo fungicidal tawm tsam I. crassa.
Cov haujlwm no tau txhawb los ntawm Lavxias Lub Tsev Haujlwm rau Kev Tshawb Fawb Qib Ib Nrab (Nyiaj Pab No. 20-016-00139).
Cov ntsiab lus tau luam tawm hauv phau ntawv xov xwm "Kev Tiv Thaiv Cov Lus Cog Tseg", 2020, 103 (3)