CUN Chudinov, V.A. Platonov, A.V. Alexandrova, S.N. Elansky
Dhawaan ayaa la muujiyay in fungus ascomycete Ilyonectria crassa ay awood u leedahay in ay qaadsiiso digriiga baradhada. Shaqadani waa tii ugu horreysay ee lagu falanqeeyo astaamaha bayoolojiga iyo iska caabbinta qaar ka mid ah fungicides-ka I. cilladda muruqsan ee laga soocay baradhada. Isku xigxiga noocyada gaarka ah ee gobollada ee "baradhada" culeyska ayaa la jaanqaaday kuwii hore loo helay ee fungi laga soocay xididdada daffodil, ginseng, aspen iyo beech, nalalka ubaxyada iyo caleemaha tulip. Sida muuqata, dhir badan oo duurjoog ah iyo kuwa beero ah ayaa kayd u noqon kara I. crassa. Nooca la baaray ee yaanyada iyo xaleef baradho qaba, laakiin ma uusan qaadsiinin miraha yaanyo oo dhan iyo tuber baradhada oo aan wali taaban. Tani waxay muujineysaa in I. crassa uu yahay dulin dhaawac. Qiimeynta iska caabbinta fludioxonil, difenoconazole iyo azoxystrobin oo leh dhexdhexaad nafaqo ayaa muujisay waxtarka sare ee daawooyinkan.
Heerka EC50 (isku-ururinta fungicide, oo hoos u dhigta 2 jeer heerka koritaanka shucaaca ee gumeysiga marka loo eego xakamaynta fungicidal) wuxuu u dhigmaa 0.4; 7.4 iyo 4 mg / l, siday u kala horreeyaan. Suurtagalnimada horumarinta cudurka ee uu sababay I. crassa waa in la tixgeliyaa marka qiimeynta phytopathological ee digriiga baradhada iyo horumarinta tallaabooyinka ilaalinta dhirta.
Horumarinta noolaha phytopathogenic wuxuu keenaa khasaaro badan dhammaan heerarka koritaanka iyo keydinta baradhada. Markii la qorsheynayo tallaabooyinka ka hortagga, sida caadiga ah, cudurada sida gaarka ah loo yaqaan ayaa la tixgeliyaa, sida noocyada hiddo-wadayaasha Alternaria, Fusarium, Phoma, Helminthosporium, Colletotrichum, Phytophthora, iwm. Si kastaba ha noqotee, sanadihii la soo dhaafay, warbixino aad iyo aad u badan ayaa ka soo muuqday muuqaalka microorganisms-ka cusub ee phytopathogenic on baradhada. Bayoolajiyadooda si liidata ayaa loo bartay, waxtarka sunta loo isticmaalo baradhada ee la xiriirta iyaga lama garanayo, hababka ogaanshaha cudurka lama soo saarin. Iyada oo leh horumarin ballaaran, waxay awood u leeyihiin inay waxyeello weyn u geystaan dalagga baradhada. Mid ka mid ah noolaha noocan ah waa fungus ascomycete Ilyonectria crassa (Wollenw.) A. Cabral & Crous, oo markii ugu horreysay ay ku arkeen qorayaashu digriiga baradhada (Chudinova et al., 2019).
Shaqadani waxay soo bandhigeysaa natiijooyinka falanqaynta I. cilladda cadaadiska ee ka go'day digriiga baradhada. Qaab-dhismeedka qaab-dhismeedka iyo qaab-dhismeedyada kala duwan ee I. crassa, isku-xigxigyada Nucleotide ee gobollada DNA-da u gaarka ah noocyada kala duwan, u nugul baradhada iyo yaanyada, iyo iska caabbinta qaar ka mid ah fangasyada caanka ah ayaa la darsay.
Qalabka iyo hababka
Waxaan u adeegsanay culeyska I. crassa 18KSuPT2 oo lagu go'doomiyay 2018 ka soo qaadidda tuber baradho cudurka qaba oo ku koray gobolka Kostroma. Qaaxada waxaa saameyn ku yeeshay nooca qurmay ee qalalan oo leh dalool daboolan mycelium bunni fudud Iyada oo la adeegsanayo irbad kala-firidhsan, fungal mycelium ayaa loo wareejiyay weel Petri ah oo leh agar dhexdhexaad ah (biirka wort 10%, agar 1.5%, penicillin 1000 U / ml). Taarikada waxaa lagu geliyay mugdiga 24 ° C.
A Leica DM2500 microscope light oo wata kaamirada dhijitaalka ah ee loo yaqaan 'ICC50 HD camera' iyo Leica M80 microscope 'binocular microscope' oo leh kaamirada dhijitaalka ah ee IC80HD (Leica Microsystems, Germany) ayaa loo isticmaalay in lagu sawiro, lagu qiimeeyo cabirka iyo qaab dhismeedka isboortiga iyo xubnaha jirka.
Si loo kala saaro DNA, fangaska 'mycelium' wuxuu ku koray dhexdhexaad digir dhexdhexaad ah, ka dibna waxaa lagu qaboojiyay nitrogen dareere ah, la isku daray, oo la geliyay keydka CTAB, oo lagu nadiifiyey chloroform, laba jeerna lagu dhaqay 2% aalkolo.
Habka soo saarida DNA waxaa si faahfaahsan loogu sharaxay maqaalka Kutuzova et al. (2017).
Si loo go'aamiyo noocyada hababka moodeelka loona barbardhigo noocyada kale ee caanka ah ee loo yaqaan 'crassa', PCR waxaa lagu sameeyay horudhac u oggolaanaya ballaadhinta noocyada u gaarka ah gobollada DNA: ITS1-5,8S-ITS2 (hordhaca ITS5 / ITS4, White et al., 1990), gobollada hidda -tubulin (Bt2a / Bt2b, Glass, Donaldson, 1995) iyo tarjumaadda kordhinta qodobka 1α (tef1α) (hordhacayaasha EF1-728F / EF1-986R, Carbone iyo Kohn, 1999). Amplicons dhererka la rabay ayaa laga soo saaray jeelka iyadoo la isticmaalayo xirmada 'Evrogen CleanUp'. Gobollada la kordhiyay ayaa la isku xigxigay iyadoo la adeegsanayo Qalabka 'BigDye® Terminator v3.1 Cycle Sequencing Kit' (Applied Biosystems, CA, USA) oo lagu sameeyay Applied Biosystems 3730 xl automated sequencer (Applied Biosystems, CA, USA). Isku xigxiga Nucleotide-ka soo baxa ayaa loo isticmaalay in lagu raadiyo ciyaar ku jirta keydka xogta GenBank ee Xarunta Qaranka ee Macluumaadka Bayoolajiyadda Mareykanka (NCBI) Falanqaynta fayoolajiyada waxaa lagu sameeyay iyadoo la adeegsanayo barnaamijka MEGA 6 (Tamura et al., 2013).
Go'aan ka gaadhista faafida ayaa lagu fuliyay miraha cagaaran oo dhan ee yaanyo waaweyn oo midho badan leh (noocyo kala duwan oo Dubrava ah) iyo digriiga baradhada (noocyada kala duwan ee Gala). Intaas waxaa sii dheer, si loo ekeysiiyo burburka miraha iyo digriiga waxyeeloobay, waxaan isticmaalnay xaleef isku miro ah iyo digriiga. Xaleefyada digriiga ayaa lagu meeleeyay qolal qoyan, kuwaas oo ahaa suxuunta Petri oo warqad miiraha qoyan salka ugu jirta. Isbarbar dhig ayaa la dul dhigay warqadda, taas oo, markeeda, xaleefyo digriiga ah ama miraha la saaray. Digriiga dhan iyo miraha ayaa sidoo kale lagu meeleeyay weel ay ku jiraan warqad miiraha qoyan salka. Badhtamaha jeex (ama dusha sare ee tuber ama midho), gabal agar (5 × 5 mm) oo leh fungal hyphae ayaa la dhigay 5 maalmood ka dib markii uu ku soo kordhayay wort agar.
Qiimeynta iska caabinta noocyada fangas ee fungicides waxaa lagu sameeyay xaaladaha shaybaarka ee agar nafaqada dhexdhexaadka ah. Waxaan baranay u nuglaanta daawooyinka fungicidal Maxim, KS (walxaha firfircoon ee fludioxonil, 25 g / l), Quadris, KS (azoxystrobin 250 g / l), Scor, EC (difenoconazole 250 g / l) (Buugga Gobolka ..., 2020). Qiimeyn ayaa lagu sameeyay suxuunta Petri ee wort-agar medium iyada oo lagu daray daawooyinka la baray ee uruurinta walaxda firfircoon ee 0.1; mid; 1 ppm (mg / L) (loogu talagalay fludioxonil iyo difenoconazole), 10; toban; 1 ppm (loogu talagalay azoxystrobin) iyo warbaahinta oo aan lahayn fungicide (xakameyn). Sunta loo yaqaan "fungicide" ayaa lagu daray dhalaalay oo la qaboojiyey ilaa 10 ° C dhexdhexaad ah, ka dib markii taas oo dhexdhexaad ah lagu shubay saxamada Petri. Xannibaadda agar oo leh fungal mycelium ayaa la dhigay bartamaha saxanka Petri waxaana lagu carbiyey heerkulka 100 ° C mugdiga. Ka dib 60 maalmood oo ku-qaadis ah, dhexroorrada gumeysiyada waxaa lagu cabiray laba jiho oo iswaafajinaya; natiijooyinka cabbirka ee gumaysi kasta celcelis ahaan. Tijaabooyinka waxaa lagu sameeyay seddex nooc. Iyada oo ku saleysan natiijooyinka falanqaynta, EC24 ayaa la xisaabiyay, oo u dhiganta isku urursanaanta fungicide, taas oo kala badh heerka kororka radial ee gumeysiga marka loo eego xakamaynta fungicidal.
Natiijooyinka iyo dood
On suxuunta petri leh wort agar, fungus la sameeyay gumeysi leh mycelium flocculent cad. Dhexdhexaadiyaha ka hooseeya mycelium wuxuu isu beddelay casaan-bunni. Markuu dhexdhexaadku qallalo, fangaska wuxuu sameeyay kudka laba nooc oo isku mid ah iyo isku darka conidiophores ee sporodochia yar. Macroconidia waa dherer, cylindrical, oo leh hal ilaa saddex septa, celcelis dherer 27.2 µm oo leh qiyam kala duwan 23.2 ilaa 32.2 µm, ballac - ilaa 4.9 µm (Sawirka 1). Celceliska dhererka microconidia waa 14.3 µm oo leh qiimayaal kala duwan oo ah 10.3 ilaa 18.1 µm, ballacuna waa ilaa 4.0 µm. Dhammaan astaamaha macro-iyo micromphophological waxay ku habboon yihiin kala duwanaanta noocyada noocyada Ilyonectria crassa (Cabral et al., 2012).
Isku xigxiga noocyada gaarka ah ee DNA-da (ITS, b-tubulin, TEF 1α) waxay si buuxda isugu beegmeen taxanaha I. muruqyada muruqyada ee aan horay u baranay (Chudinova et al., 2019, Shaxda 1). Si loo barto baahsanaanta I. crassa ee gobollada kale iyo in la falanqeeyo muuqaalka dhaqammada ay saameysey, isku-xigxigyada DNA ee ku jira keydka xogta ee GenBank ayaa la falanqeeyey (Shaxda 1). Isukeenku wuxuu ahaa 86 ilaa 100%. Isku xigxiga dhammaan seddexda gobol ee DNA ee "baradhada" I. cilladda 'koro' waxay la mid tahay taxanaha noocyada ka soocan guluubka lily iyo xididdada narcissus ee Nederland iyo xididka ginseng ee Kanada. Waxaan ku guuldareysanay inaan helno noocyo kale oo kala ah I. crassa oo leh seddex falanqeyn isku mid ah oo la mid ah keydka macluumaadka. Si kastaba ha noqotee, falanqaynta isku-darka ITS iyo b-tubulin isku xigxiga ayaa muujiyay joogitaanka I. crassa ee caleemaha tulip ee Boqortooyada Midowday. Fungi oo leh taxane ITS ah oo isku mid ah ayaa lagu aqoonsaday falanqaynta mycobiota ee xididdada aspen ee Kanada iyo xididdada beech ee Talyaaniga, digriiga baradhada ee Saudi Arabia (Shaxda 1). Natiijooyinka daraasaddan waxay muujineysaa in I. crassa ay leedahay qeybis caalami ah ayna awood u leedahay in ay qaadsiiso noocyada dhirta kala duwan.
Markii la go'aaminayey jirrada ku jirta xaleefyada yaanyada iyo baradhada maalinta 5aad, dhexroorka nabarku wuxuu gaadhay 1.5 cm Culayska la baaray ma qaadsiinin miraha yaanyo oo dhan iyo tuber baradho ah oo aan wali taaban. Si kastaba ha noqotee, sepals-ka ayaa saameyn ku yeeshay yaanyada. Si looga saaro suurtagalnimada faddarayn ka timaadda mycelium-ka lagu sameeyay jeexitaanka tuber-ka baradho, go'doomin fangas ah ayaa lagu go'doomiyay dhaqan saafi ah. Waxay gebi ahaanba la mid ahayd culeyska waalidka. Sida muuqata, I. crassa waa dulin nabar.
Daaweynta pre-beerista ee digriiga abuurka leh fungicides yaraynaysaa horumarinta cudurada on dhirta inta lagu guda jiro xilli sii kordhaya. Xulashada sunta fangaska wax ku oolka ah, waxaa muhiim ah in la qiimeeyo midkee waxtar u leh I. сrassa. Shaqadu waxay baratay walxaha firfircoon ee baahsan ee fungicides - fludioxonil, azoxystrobin, difenoconazole. Fludioxonil waxay ku jirtaa dhowr isku dar ah oo loo adeegsado labiska miraha iyo digriiga abuurka ka hor inta aan la beerin. Fludioxonil (Maxim) sidoo kale waxaa loo isticmaalaa in lagu daaweeyo digriiga abuurka ka hor kaydinta. Difenoconazole iyo azoxystrobin ayaa sidoo kale lagu daray dhowr diyaargarow oo loo adeegsaday soo saarista walxaha abuurka, iyo sidoo kale u diyaargarowga loogu talagalay ka shaqeynta dhirta daaqsinka ah (Buugga Gobolka ..., 2020).
Qiyaasta koritaanka I. cilsa ayaa lagu bartay warbaahinta (Jaantus. 2) oo leh uruurinta kaladuwan ee walxaha firfircoon: fludioxonil (EC50 = 0.4 ppm), azoxystrobin (EC50 = 4 ppm), iyo difenoconazole (EC50 = 7.4 ppm) (Shaxda 2). Diyaargarowyadan waxaa loo qaadan karaa inay yihiin kuwo wax ku ool ah marka loo eego I. crassa, maadaama EC50-kooda uu aad uga hooseeyo ka-fiirsashada lagula taliyay ee diyaarinta dareeraha shaqeeya ee loo isticmaalo daweynta digriiga. Sida ku cad Catalog-ka Gobolka ... (2020), isku-uruurinta fludioxonil ee dareeraha lagu daaweeyo digriiga digrii waa min 500 ilaa 1000 ppm, azoxystrobin (dareeraha lagu daaweynayo salka hoose ee dhogorta) - 3750-9375 ppm, difenoconazole (dareeraha daaweynta dhirta daaqsinka ah) - 187.5 - 625 ppm.
Jadwalka 1. Isku ekaanshaha taxanaha noocyada gaarka ah ee taxadarka culeyska 18KSuPT2 iyo noocyada cilladda Ilyonectria crassa oo laga heli karo keydka xogta Genbank
Cadaadiska | Warshad martida loo yahay, goobta qashin-saarka | Tirooyinka isku xigxiga ee lagu shubay GenBank, boqolkiiba isbarbardhiga | link | ||
ITS | β-tubulin | TEF 1α | |||
17KSPT1 iyo 18KSuPT2 | Digriiga baradhada, gobolka Kostroma | MH818326 | MH822872 | MK281307 | Chudinova et al., 2019, shaqadan |
CBS 158/31 | Narcissus xididdada, Nederland | JF735276 100 | JF735394 100 | JF735724 99.3 | Cabral iyo al., 2012 |
CBS 139/30 | Nalalka Lily, Netherlands | JF735275 100 | JF735393 99.7 | JF735723 99.3 |
|
NSAC-SH-1 | Xididka Ginseng, Kanada | AY295311 99.4 | JF735395 100 | JF735 / 725 99.6 |
|
RHS235138 | Caleen Tulip, UK | KJ475469 100 | KJ513266 100 | ND | Denton, Denton, 2014 |
MT294410 | Xididdada Aspen, Kanada | MT294410 100 | ND | ND | Ramsfield iyo al., 2020 |
ER1937 | Beech, Talyaaniga | KR019363 99.65 | ND | ND | Tizzani, Haegi, Motta. Soo gudbin toos ah |
KAUF19 | Digriiga baradhada, Sacuudi Carabiya | HE649390 98.3 | ND | ND | Gashgari, Gherbawy, 2013 |
ND = lama dhigin
Jadwalka 2. Iska caabbinta cilladda Ilyonectria ee sunta lagu buufiyo
(walax firfircoon) | EC50, ppm | ||||
Maalinta 3 | Maalinta 5 | Maalinta 7 | |||
Xakamaynta | 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 |
Shaqadeenna, I. noocyada 'crassa' ayaa laga go'doomay digriiga baradhada ee gobollada Kostroma iyo Moscow (Chudinova et al., 2019). Qayb aad u saraysa oo ah noocyada fungal ee leh taxanaha ITS ee isku midka u ah aniga iyo rrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrr - fg fungi ah oo leh taxane ITS oo la mid ah aniga. Sida muuqata, I. crassa ugama badna baradhada sida ay umuuqato. Tijaabooyinkayagu waxay muujiyeen in fangaska uu ku dhici karo miraha yaanyada ee waxyeelada soo gaadhay. Waxaa laga ogyahay suugaanta in I. crassa uu awood u leeyahay inuu ku kobco carrada saprotrophically (Moll et al., 2013), iyo sidoo kale saameyn ku yeesho dhir kala duwan, xitaa kuwa canshuuraha ka fog sida daffodils, ubaxyada, ginseng, aspen, iyo beech (Shaxda 2016). mid). Sida muuqata, dhir badan oo duurjoog ah iyo kuwa beero ah ayaa kayd u noqon kara I. crassa. Waxyaabaha kor ku xusan waxay muujinayaan in marka la horumarinayo tallaabooyinka ilaalinta, ay lagama maarmaan tahay in la tixgeliyo suurtagalnimada in la saameeyo digriiga baradhada leh funguskan. Diyaargarow ballaaran oo loogu talagalay daaweynta digriiga baradhada ay ku jiraan fludioxonil, azoxystrobin iyo difenoconazole waxay muujiyeen waxtarka fangaska ee aadka u saraysa.
Shaqadan waxaa taageeray Hay'adda Ruushka ee Cilmi-baarista Aasaasiga ah (Grant No 20-016-00139).
Maqaalka waxaa lagu daabacay joornaalka "Warsidaha Difaaca Dhirta", 2020, 103 (3)