rie. Chudinova, V.A. Platonov, A.V. Alexandrova, S.N. Elansky
Na nso nso a, e gosiri na ascomycete ero Ilyonectria crassa nwere ike ịwakpo nduku tubers. Ọrụ a bụ nke mbụ nyochara njirimara ndu na iguzogide ụfọdụ fungicides nke nje I. crassa dịpụrụ adịpụ na poteto. Usoro nke mpaghara-kpọmkwem ụdị nke ụdị "nduku" dabara na nke a na-enwetabu maka fungi dịpụrụ adịpụ na mgbọrọgwụ nke narcissus, ginseng, aspen na beech, lily bulbs na tulip epupụta. N'ụzọ doro anya, ọtụtụ ọhịa na ubi ubi nwere ike idobere maka I. crassa. Ụdị ahụ a mụrụ butere iberi tomato na nduku, mana ebuteghị otu mkpụrụ tomato dum ma ọ bụ tuber nduku na-emebibeghị. Nke a na-egosi na I. crassa bụ ọnya ọnya. Nlele nke iguzogide fludioxonil, difenoconazole na azoxystrobin na ihe na-edozi ahụ gosipụtara ịdị irè nke ọgwụ ndị a dị elu.
Ihe ngosi EC50 (ntinye ọgwụ na-egbu egbu nke na-ebelata site na ugboro 2 ọnụego nke uto radial nke ógbè ahụ metụtara njikwa na-abụghị nke fungicides) ruru 0.4; 7.4 na 4 mg / l, n'otu n'otu. Ekwesịrị iburu n'uche ohere nke ịmalite ọrịa nke I. crassa kpatara mgbe phytopathologically na-enyocha tubers nduku na ịmepụta usoro nchebe osisi.
Mmepe nke microorganisms phytopathogenic na-eduga na mfu dị elu na ọkwa niile nke ịkọ nduku na nchekwa. Mgbe ị na-eme atụmatụ ihe nchebe, dị ka a na-achị, a na-echebara ihe ndị na-akpata ọrịa ndị a ma ama, dị ka ụdị nke genera Alternaria, Fusarium, Phoma, Helminthosporium, Colletotrichum, Phytophthora, wdg. Otú ọ dị, n'ime afọ ndị na-adịbeghị anya, akụkọ na-arịwanye elu na-apụta. banyere ọdịdị ọhụrụ phytopathogenic microorganisms na poteto. A mụọla usoro ihe ọmụmụ ha nke ọma, amabeghị uru nke fungicides ejiri na poteto megide ha, na emebebeghị usoro nyocha. Mgbe ha na-etolite n'ọha, ha nwere ike ịkpata mmebi dị ukwuu na ihe ọkụkụ nduku. Otu n'ime microorganisms ndị a bụ fungus ascomycete Ilyonectria crassa (Wollenw.) A. Cabral & Crous, nke ndị edemede mbụ chọpụtara na tubers nduku (Chudinova et al., 2019).
Akwụkwọ a na-egosi nsonaazụ nke nyocha nke nsogbu I. crassa dịpụrụ adịpụ na tuber nduku. A mụrụ usoro ọmụmụ nke colonies na mycelial Ọdịdị nke I. crassa, nucleotide usoro nke ụdị-kpọmkwem DNA ngalaba, virulence na poteto na tomato, na iguzogide ụfọdụ fungicides na-ewu ewu.
Akụrụngwa na ụzọ
Ụdị a na-eji na-arụ ọrụ a bụ I. crassa 18KSuPT2, dịpụrụ adịpụ na 2018 site na tuber nduku butere na mpaghara Kostroma. The tuber na-emetụta akọrọ ire ere na oghere kpuchie na ìhè aja aja mycelium. N'iji agịga dissecting na-adịghị mma, a na-ebufe mycelium fungal ahụ n'ime efere Petri nke nwere agar (beer wort 10%, agar 1.5%, penicillin 1000 units/ml). A na-etinye efere ndị ahụ n'ọchịchịrị na 24 Celsius.
Maka ịse foto, a na-eji nyocha nke nha na ọdịdị ọdịdị nke spores na sporulation, a na-eji microscope ìhè Leica DM2500 nwere igwefoto dijitalụ ICC50 HD na mikroskopu binocular Leica M80 nwere igwefoto dijitalụ IC80HD (Leica Microsystems, Germany).
Iji kewapụ DNA, a na-akụ mycelium fungal ahụ n'ime mmiri mmiri mmiri, wee kpoo ya na mmiri mmiri nitrogen, megharịa ya, tinye ya na nchekwa CTAB, sachaa ya na chloroform, wee jiri mmanya 2% sachaa ugboro abụọ.
A kọwara usoro mmịpụta DNA n'ụzọ zuru ezu n'isiokwu nke Kutuzova et al. (2017).
Iji chọpụta ụdị njirimara site na usoro mkpụrụ ndụ yana atụnyere ụdị ndị ọzọ ama ama nke I. crassa, a na-eme PCR na primers nke kwere ka mmụba nke mpaghara DNA dị iche iche: ITS1-5,8S-ITS2 (primers ITS5 / ITS4, White et al., 1990), mpaghara b genes -tubulin (Bt2a/Bt2b, Glass na Donaldson, 1995) na ntụgharị elongation factor 1α (tef1α) (primers EF1-728F/EF1-986R, Carbone na Kohn, 1999). A na-ewepụta amplicons nke ogologo a chọrọ na gel site na iji ngwa CleanUp sitere na Evrogen. A na-ahazi mpaghara ndị a gbanyere amụba n'usoro site na iji BigDye®Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, CA, USA) na Applied Biosystems 3730 xl automated sequencer (Applied Biosystems, CA, USA). Ejiri usoro nucleotide enwetara wee chọọ egwuregwu na nchekwa data GenBank nke US National Center for Biotechnology Information (NCBI). Emere nyocha phylogenetic site na iji mmemme MEGA 6 (Tamura et al., 2013).
A na-eme mkpebi nke virulence na mkpụrụ osisi akwụkwọ ndụ akwụkwọ ndụ zuru oke nke tomato buru ibu (ụdị Dubrava) na tubers nduku (ụdị Gala). Na mgbakwunye, iji mebie mmebi nke mkpụrụ osisi na tubers mebiri emebi, a na-eji mpekere nke otu mkpụrụ osisi na tubers mee ihe. A na-etinye mpekere tuber n'ime ụlọ nwere iru mmiri, nke bụ efere Petri nwere akwụkwọ nzacha mmiri na ala. A na-etinye ihe mmịfe iko na akwụkwọ ahụ, nke, n'aka nke ya, a na-etinye mpekere nke tubers ma ọ bụ mkpụrụ osisi. A na-etinyekwa tubers na mkpụrụ osisi n'ime akpa nwere akwụkwọ nzacha mmiri na ala. A na-etinye otu agar (5 × 5 mm) nke nwere hyphae fungal n'etiti iberi (ma ọ bụ n'elu tuber ma ọ bụ mkpụrụ osisi ahụ) na hyphae fungal mgbe ụbọchị 5 gachara na wort agar.
Achọpụtara nguzogide nke ụdị fungal na fungicides na ọnọdụ ụlọ nyocha na usoro nri agar. Anyị mụrụ ihe gbasara ike ọgwụ fungicidal Maxim, KS (fludioxonil na-arụ ọrụ, 25 g / l), Quadris, KS (azoxystrobin 250 g/l), Skor, CE (difenoconazole 250 g/l) (Katalọgụ steeti..., 2020) ). A na-eme nyocha ahụ na nri Petri na wort-agar na mgbakwunye nke ọgwụ ndị a na-amụ na ntinye nke ihe na-arụ ọrụ nke 0.1; 1; 10 ppm (mg/l) (maka fludioxonil na difenoconazole), 1; 10; 100 ppm (maka azoxystrobin) na na a na-enweghị fungicides (njikwa). A na-agbakwunye fungicides na agbaze ma mee ka ọ dị jụụ ruo 60 Celsius C, mgbe nke ahụ gasịrị, a na-awụsa ihe na-edozi ahụ n'ime efere Petri. A na-etinye ihe mgbochi agar nwere mycelium fungal n'etiti efere Petri ma kụọ ya na okpomọkụ nke 24 Celsius C n'ọchịchịrị. Mgbe ụbọchị 7 nke nnabata gasịrị, a tụrụ dayameta nke ógbè ndị ahụ n'akụkụ abụọ na-agbanwe agbanwe; A na-agbakọ nsonaazụ nha maka mpaghara ọ bụla. Emere nnwale ugboro atọ. Dabere na nsonaazụ ule ahụ, a gbakọrọ index EC50, hà nhata na mkpokọta fungicides nke belatara ọnụ ọgụgụ nke uto radial nke ógbè ahụ na njikwa na-abụghị nke fungicides.
Nsonazụ na mkparịta ụka
Na efere Petri na wort agar, ero ahụ hibere obodo nwere mycelium na-acha ọcha. Ndị na-ajụ n'okpuru mycelium tụgharịrị uhie-agba aja aja. Mgbe ọkara akpọnwụọ, ero ahụ kpụrụ ụdị spores abụọ na otu conidiophores na ndị a chịkọtara n'ime obere sporodochium. Macroconidia bụ elongated, cylindrical, na otu na-atọ septa, nkezi ogologo 27.2 µm na a nso nke ụkpụrụ si 23.2 ka 32.2 µm, obosara - ruo 4.9 µm (Fig. 1). Nkezi ogologo nke microconidia bụ 14.3 µm nwere oke ụkpụrụ sitere na 10.3 ruo 18.1 µm, obosara - ruo 4.0 µm. Ihe odide macro na micromorphological niile na-adaba n'ụdị dịgasị iche iche nke ụdị Ilyonectria crassa (Cabral et al., 2012).
Usoro nke mpaghara DNA nke ụdị-kpọmkwem (ITS, b-tubulin, TEF 1α) dabara kpamkpam na usoro nke strains I. crassa anyị mụrụ na mbụ (Chudinova et al., 2019, Tebụl 1). Iji mụọ mgbasa nke I. crassa na mpaghara ndị ọzọ ma nyochaa ụdị dị iche iche nke ihe ọkụkụ emetụtara, a na-enyocha usoro DNA yiri nke ahụ na nchekwa data GenBank (Table 1). Mgbakwunye ahụ sitere na 86 ruo 100%. Usoro nke mpaghara DNA atọ nke ụdị "potato" nke I. crassa yiri usoro nke ụdị dịpụrụ adịpụ na lily bulbs na mgbọrọgwụ narcissus na Netherlands na site na mgbọrọgwụ ginseng na Canada. Anyị enweghị ike ịchọta ụdị nsogbu I. crassa ọzọ na usoro atọ a tụlere na ọdụ data mepere emepe. Agbanyeghị, nyocha nke usoro ITS echekwara na b-tubulin gosipụtara ọnụnọ nke I. crassa na akwụkwọ tulip na UK. Achọpụtara fungi nwere usoro ITS yiri ya mgbe a na-enyocha mycobiota nke mgbọrọgwụ aspen na Canada na beech na Italy, na tubers nduku na Saudi Arabia (Table 1). Nsonaazụ nke ọmụmụ a na-egosi na I. crassa nwere nkesa zuru ụwa ọnụ ma nwee ike ịwakpo ụdị osisi dị iche iche.
Mgbe ị na-achọpụta ọrịa nje na mkpụrụ osisi tomato na poteto n'ụbọchị 5, dayameta nke ọnya ahụ ruru 1.5 cm n'otu oge ahụ, nsogbu a na-amụ emeghị ka mkpụrụ osisi tomato dum ma ọ bụ tuber nduku na-adịghị emebi emebi. Otú ọ dị, a hụrụ mmebi nke sepals na tomato. Iji wezuga ohere nke mmetọ, a na-ekewapụ iche iche fungal ka ọ bụrụ omenala dị ọcha site na mycelium nke tolitere na iberi nke tuber nduku. Ọ bụ kpam kpam yiri ụdị nne na nna. I. crassa dị ka ọnya nje.
Tupu akuku ọgwụgwọ nke mkpụrụ tubers na fungicides nwere ike ibelata mmepe nke ọrịa na osisi n'oge na-eto eto n'oge. Iji họrọ fungicides dị irè, ọ dị mkpa iji nyochaa nke dị irè megide I. crassa. A na-eji ihe ndị na-arụ ọrụ nke fungicides na-arụ ọrụ nke ọma - fludioxonil, azoxystrobin, difenoconazole. Fludioxonil bụ akụkụ nke ọtụtụ nkwadebe agwakọtara eji agwọ mkpụrụ na tubers mkpụrụ tupu a kụọ ya. A na-ejikwa Fludioxonil (Maxim) agwọ tubers mkpụrụ tupu echekwa ya. A na-etinyekwa Difenoconazole na azoxystrobin n'ọtụtụ nkwadebe eji eme ihe maka nhazi mkpụrụ, yana na nkwadebe maka ịhazi osisi ahịhịa (Katalọgụ steeti ..., 2020).
A mụrụ mmụba nke I. crassa na mgbasa ozi (Fig 2) na nchịkọta dị iche iche nke ihe ndị na-arụ ọrụ: fludioxonil (EC50 = 0.4 ppm), azoxystrobin (EC50 = 4 ppm) na difenoconazole (EC50 = 7.4 ppm) (Table 2) . Enwere ike iwere ọgwụ ndị a dị irè megide I. crassa, ebe ọ bụ na EC50 ha dị ntakịrị karịa ntinye ọgwụ akwadoro na mmiri na-arụ ọrụ nke a na-eji agwọ tubers. Dabere na katalọgụ steeti ... (2020), ntinye nke fludioxonil na mmiri mmiri maka ịgwọ tubers nduku sitere na 500 ruo 1000 ppm, azoxystrobin (n'ime mmiri mmiri maka ịgwọ ala furrow) - 3750-9375 ppm, difenoconazole. (n'ime mmiri mmiri maka ịgwọ ahịhịa ahịhịa) - 187.5-625 ppm.
Isiokwu 1. Usoro myirịta n'etiti ụdị-kpọmkwem usoro nke nje 18KSuPT2 na Ilyonectria crassa strains dị na Genbank nchekwa data.
Nje | Osisi ọbịa, ebe excretion | Nọmba usoro edobere na GenBank, pasent nke myirịta | njikọ | ||
ISI | β-tubulin | TEF 1a | |||
17KSPT1 na 18KSuPT2 | Nduku tuber, Kostroma mpaghara. | MH818326 | MH822872 | MK281307 | Chudinova et al., 2019, ọrụ a |
CBS 158/31 | Narcissus mgbọrọgwụ, Netherlands | JF735276 100 | JF735394 100 | JF735724 99.3 | Cabral et al., 2012 |
CBS 139/30 | Lily bulb, Netherlands | JF735275 100 | JF735393 99.7 | JF735723 99.3 |
|
NSAC-SH-1 | Ginseng mgbọrọgwụ, Canada | AY295311 99.4 | JF735395 100 | JF735/725 99.6 |
|
RHS235138 | Akwụkwọ Tulip, UK | KJ475469 100 | KJ513266 100 | ND | Denton, Denton, 2014 |
MT294410 | Aspen mgbọrọgwụ, Canada | MT294410 100 | ND | ND | Ramsfield et al., 2020 |
ER1937 | Buc, Italy | AKA019363 99.65 | ND | ND | Tizzani, Haegi, Motta. Nrubeisi ozugbo |
KAUF19 | Nduku tuber, Saudi Arabia | HE649390 98.3 | ND | ND | Gashgari, Gherbawy, 2013 |
ND = etinyeghị ya
Tebụl 2. Nguzogide Ilyonectria crassa na fungicides
(ihe na-arụ ọrụ) | EC50, ppm | ||||
Ụbọchị 3 | Ụbọchị 5 | Ụbọchị 7 | |||
Njikwa | 17 ± 2 | 33 ± 5 | 47 ± 3 | ||
Quadris, KS (fzoxystrobin) | 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 | |||
Akara, EC (difenoconazole) | 18 ± 1 | 35 ± 2 | 48 ± 1 | ||
11 ± 1 | 24 ± 3 | 35 ± 4 | |||
11 ± 1 | 13 ± 1 | 17 ± 3 |
N'ime ọrụ anyị, eriri I. crassa dịpụrụ adịpụ site na tubers nduku na mpaghara Kostroma na Moscow (Chudinova et al., 2019). Achọpụtara ọnụ ọgụgụ dị elu nke ụdị fungal na usoro ITS yiri nke I. crassa mgbe a na-enyocha mycobiota nke tubers nduku na Saudi Arabia (Gashgari na Gherbawy, 2013). N'ụzọ doro anya, I. crassa adịghị ụkọ na poteto dị ka ọ pụrụ iyi. Nnwale anyị mere gosiri na ero nwere ike bute mkpụrụ osisi tomato mebiri emebi. A maara site na akwụkwọ akụkọ na I. crassa nwere ike ịmepụta saprotrophically na ala (Moll et al., 2016), ma na-ebutekwa ụdị osisi dịgasị iche iche, ọbụna ndị dị anya na okwu taxonomic dị ka daffodils, lilies, ginseng, aspen, na beech (Table 1). N'ụzọ doro anya, ọtụtụ ọhịa na ubi ubi nwere ike idobere maka I. crassa. Ihe dị n'elu na-egosi na mgbe ị na-emepụta ihe nchebe, ọ dị mkpa iburu n'uche na o nwere ike imebi tubers nduku site na ero a. Nkwadebe nke a na-ejikarị eme ihe maka ịgwọ tubers nduku, nwere fludioxonil, azoxystrobin na difenoconazole, egosila nnukwu uru fungicidal megide I. crassa.
Ọrụ ahụ kwadoro site na Russian Foundation for Basic Research (onyinye No. 20-016-00139).
E bipụtara akụkọ a n'akwụkwọ akụkọ "Bulletin of Plant Protection", 2020, 103(3)