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1.
Induction of germination of microsclerotia by exudates from plant roots may be important for the control ofV. dahliae. Laboratory experiments with root observation boxes were carried out to assess the influence of root tips of seven crop species and cultivars on the germination of microsclerotia ofVerticillium dahliae in soil under controlled conditions. The root density of crops was measured in a field experiment. The results of the laboratory experiments and the field experiment were combined to estimate the total effect of crops on the population of microsclerotia in the field. Germination of microsclerotia was stimulated by all crops compared to a control without a crop. Among crops, roots of potato cvs Element and Astarte had a larger stimulation effect on microsclerotia than that of potato Ostara, pea, flax, sugar beet or onion. The number of hyphae per microsclerotium decreased with distance from the root surface regardless of the crop species or cultivar. Differences in root densities, in the affected root zones and in the stimulation effect on germination of microsclerotia caused large differences among crops in the effect on the population of microsclerotia in the soil. However, growing a rop with the special purpose to reduce the level ofV. dahliae inoculum in the soil is an inefficient control measure, because only a small part of the total soil volume is affected by roots and the number of hyphae per microscleroium affected is too low.Abbreviations MS microsclerotia, microsclerotium  相似文献   

2.
For many soilborne plant pathogens, disease results from multiple root infections. Studying the infection dynamics of single or multiple propagules of these pathogens applied at one site of the root system may be the basis for understanding the development of disease caused by multiple root infections. The effect of single-site inoculations of roots of eggplant seedlings with microsclerotia of the wilt-causing fungusVerticillium dahliae, was studied. Experiments were conducted using specially designed pots which enabled the incorporation and removal of inoculum in the soil. Inoculations were carried out by placing microsclerotia, firmly embedded in small sections of polypropylene screen filter, directly below the growing tip of the main root of young eggplant seedlings. Three to 4 days after inoculation, the root had grown over the screen filter, and the filter was removed. Root platings showed high infection levels at the inoculation site, but also several (discrete) root infections were noted some distance above and below the site of inoculation. Exposure of the root to the lowest number of microsclerotia (26/inoculation site) was sufficient to lead to up to 65% root infections. Number of plants with root infections declined over time, ranging from a maximum of 65–100% 2–4 wk after inoculation, to 10–29% at 6–7 wk after inoculation. Apparently,V. dahliae died in nonsystemic infections after some time.  相似文献   

3.
To determine the host range ofVerticillium dahliae among the cultivated species in Crete, Greece, studies were carried out during 1992–2000. Based on disease symptoms observed on 28 vegetable and forage species grown in a field naturally infected byV. dahliae, and an extensive survey of the most common cultivated species grown under natural conditions, seven hosts belonging to four botanical families not previously reported as susceptible to Verticilliun wilt, and 12 hosts belonging to seven families new for Greece, were recorded. The worldwide new hosts are: anise (Anethum graveolens), chard (Beta vulgaris ssp.cicla), chickpea (Cicer arietinum), wild sweet pea (Lathyrus ochrus), lentil (Lens culinaris), marigold (Tagetes erecta) and vetch (Vicia sativa). These species could be infected by their hosts-of-originV. dahliae isolates during pathogenicity tests. http://www.phytoparasitica.org posting Jan. 13, 2002.  相似文献   

4.
A survey of common and uncommon weed species usually showing Verticillium wilt symptoms was carried out during 1992–2000 in Crete, Greece.Verticillium dahliae was isolated in 48 out of 182 sampled fields, in which several weed species were grown, from several locations in Oropedio, Lasithi. Altogether, 124 isolates ofV. dahliae were recovered from the vascular stem-tissue of 19 weed species, belonging to ten botanical families. Pathogenicity trials with 13 out of 19 weed species that have never been reported as hosts of the fungus, using for inoculation isolates which originated from the same weed species, resulted in infection of all of them, showing various disease symptoms. Seven weed species (Anthemis melanolepis, Cardaria draba, Convolvulus arvensis, Erodium sp.,Euphorbia helioscopia, Helminthotheca echioides andSinapis alba) are new hosts worldwide, and six additional species (Euphorbia sp.,Lactuca serriola, Raphanus raphanistrum, Sinapis arvensis, Sonchus oleraceus andTrifolium sp.) are new hosts for Greece. The most susceptible (isolation frequency: 27.9–52.8%, moderate disease severity) species were:Capsella bursa-pastoris, C. draba, Chenopodium album, Senecio vulgaris andSolanum nigrum. Less susceptible (isolation frequency: 4.8–17.8%, slight disease severity) were:Amaranthus sp.,A. melanolepis, C. arvensis, Erodium sp.,Euphorbia sp.,E. helioscopia, H. echioides, L. serriola, Malva sylvestris, R. raphanistrum, S. alba, S. arvensis, S. oleraceus andTrifolium sp. Some species —C. draba, C. album, L. serriola andS. nigrum L. — that usually showed external and vascular wilt symptoms, occasionally exhibited only reduced growth. Visible symptoms under natural field conditions in all 13 weed species that had never been reported as hosts ofV. dahliae were similar to those observed after their artificial inoculation. The fungus was not isolated fromFoeniculum vulgare ssp.piperitum, Oxalis corniculata andStellaria media, among other species. http://www.phytoparasitica.org posting Sept. 18, 2002.  相似文献   

5.
为明确西兰花轮作及其残体还田对土壤中病原菌大丽轮枝菌Verticillium dahliae微菌核数量、马铃薯黄萎病发生以及马铃薯产量的影响,分别于2017-2018年和2018-2019年在内蒙古自治区乌兰察布市凉城县岱海镇元山子农场人工病圃和锡林郭勒盟正蓝旗上都镇菜园村自然病圃开展田间试验,利用NP-10培养基测定不同处理土壤中的大丽轮枝菌微菌核数量,并对马铃薯地上叶片和地下块茎上黄萎病的发病率、病情指数以及马铃薯总产量和商品薯产量进行调查。结果显示,以马铃薯连作方式为对照,西兰花轮作及其残体还田后土壤中大丽轮枝菌微菌核的数量明显减少,其中在人工病圃于7、8和9月调查时分别降低了8.46%、36.41%和36.27%,在自然病圃于7、8和9月调查时则分别降低了7.22%、38.06%和48.84%;马铃薯地上叶片和地下块茎上黄萎病的发病率和病情指数均显著降低,在人工病圃对黄萎病的防效为32.91%和54.82%,在自然病圃对黄萎病的防效为46.72%和34.78%;马铃薯总产量和商品薯产量均有所提高,在人工病圃分别增产27.74%和40.08%,在自然病圃分别增产7.12%和8.77%。表明西兰花作为理想的轮作作物可应用于马铃薯黄萎病的有效防控。  相似文献   

6.
土壤大丽轮枝菌微菌核的快速定量检测   总被引:4,自引:0,他引:4  
 微菌核是大丽轮枝菌在土壤中的主要存活结构和黄萎病的初侵染来源。对土壤中大丽轮枝菌微菌核进行定量是黄萎病监测和预警的基础。本研究以大丽轮枝菌Internal Transcribed Spacer (ITS)区特异性引物对P1/P2扩增产物的重组质粒为标准品,构建SYBR Green I实时荧光定量PCR反应的标准曲线,结合土样水筛法建立了土壤大丽轮枝菌微菌核定量检测体系。同时,建立了土壤中微菌核数量与棉花黄萎病发病率的关系模型。结果表明,实时定量PCR检测灵敏度比常规PCR高10倍,检测下限为1个微菌核/克土,在5.54×102~5.54×107copies范围内,DNA拷贝数的对数值与Ct值具有良好的线性关系。建立的土壤中微菌核个数n与Ct值之间的关系为n=e7.3-Ct/3.905。温室人工接种微菌核数量与棉花黄萎病发病率间的线性关系为y=2.710n+0.251。  相似文献   

7.
刘海洋  王伟  张仁福  姚举 《植物保护》2023,49(4):276-283
为了解北疆棉花黄萎病的发生情况及发病与土壤中黄萎病菌微菌核数量的关系和病原菌种群类型, 2021年对北疆石河子?奎屯?博乐等8市(县)棉田棉花黄萎病发病率?土壤中黄萎病菌的微菌核数量?菌株种群类型进行了抽样调查?结果表明, 北疆未发生棉花黄萎病的棉田占49.2%, 0%<发病率<5.0%的棉田占32.7%, 发病率≥5.0%的棉田占18%?与2013年?2015年相比, 2021年无病田率分别增加17.7和12.7百分点, 发病率≥5%的棉田分别减少15.7和21.6百分点?从棉田黄萎病发病率与土壤中微菌核数量的关系来看, 整体上北疆棉田棉花黄萎病发病率与微菌核数量相关性不显著(r=0.119 1); 分区域看, 石河子-沙湾片区?奎屯-乌苏片区?精河-博乐片区棉田黄萎病发病率与微菌核数量的相关系数分别为0.033 2?0.007 6?0.062 3, 均无显著相关性; 而呼图壁-玛纳斯片区棉田黄萎病发病率与微菌核数量的相关系数为0.635 7, 呈中度正相关?土壤中的黄萎病菌菌株以菌核型为主, 占57.9%, 菌丝型占23.2%, 中间型占18.9%?用特异性引物进行PCR检测表明, 土壤中黄萎病菌落叶型菌株占97.6%, 占绝对优势?本研究将为北疆棉区棉花黄萎病的综合防控提供理论依据?  相似文献   

8.
检疫性轮枝菌及其近似种的鉴定   总被引:2,自引:0,他引:2  
 大丽轮枝菌(Verticillium dahliae)和黑白轮枝菌(V. albo-atrum)在世界范围内引起多种作物的黄萎病,属于我国重要进境植物检疫对象。本研究对采自我国部分地区和CBS保存的多种植物病原性轮枝菌,包括黑白轮枝菌、大丽轮枝菌及其变种大丽轮枝菌长孢变种(V. dahliae var. longisporum)、三体轮枝菌(V. tricorpus)、变黑轮枝菌(V. nigrescens)和云状轮枝菌(V. nubilum),采用生物学特性观察,结合rDNA-ITS序列分析的方法,进行了比较和分析。结果表明:不同种类轮枝菌在休眠结构形态上具有一定差异,部分菌株不产生任何休眠结构。各供试菌株在15~25℃范围内均可生长,但黑白轮枝菌在30℃下生长受到强烈抑制,而其他菌株受影响较小。对供试菌株rDNA-ITS序列分析结果表明植物病原性轮枝菌可聚为9个分支,包括三体轮枝菌、变黑轮枝菌、云状轮枝菌、V. theobromae、大丽轮枝菌、大丽轮枝菌长孢变种和3个不同的黑白轮枝菌分支,黑白轮枝菌、大丽轮枝菌及其长孢变种亲缘关系较近。采用生物学性状结合rDNA-ITS序列分析能够更加有效地将两种检疫性轮枝菌从其他植物病原性轮枝菌中区分出来。  相似文献   

9.
K. Elena 《Phytoparasitica》2000,28(2):115-120
Vegetative compatibility among 17 isolates ofVerticillium dahliae obtained from watermelon, originating from eight regions of Greece, was investigated using complementation tests between nitrate-nonutilizing(nit) mutants. Among 529 chlorate-resistant sectors obtained, only 107 werenit mutants. These mutants were paired with tester strains (from Greece and other countries) of previously described vegetative compatibility groups (VCGs), and also were paired in many combinations among themselves. All isolates were self-compatible. Sixteen isolates were found to belong to VCG2. Only isolate V75 could not be assigned to a VCG, because the threenit mutants obtained from it showed negative reactions with the tester strains of four VCGs and with complementary mutants from other isolates. Based on this sample, we conclude that the population ofV. dahliae from watermelon in Greece is homogeneous in respect to VCG.  相似文献   

10.
Tsror  Leah  Livescu  L.  Nachmias  A. 《Phytoparasitica》1990,18(4):331-339
The tolerance response to Verticillium in potato is affected by external abiotic and biotic factors. Short-day photoperiod enhanced disease level in the susceptible cvs. ‘Nicola’ and ‘Maris Bard’, as well as in the tolerant cvs. ‘Cara’ and ‘Désirée‘. This was observed under field conditions, where plant reaction in the spring was compared with that in autumn. In some cases the disease was more pronounced in autumn, with its shorter days, than in spring, when the days are longer. The effect was measured by disease symptom severity, fungus colonization in the plants, plant height and final yield. This disease enhancement was more pronounced under controlled growth room conditions, where the only different parameter was illumination. A short photoperiod of 8 h increased the disease level relative to 16 h of light.  相似文献   

11.
为明确效应蛋白VdSRP2在大丽轮枝菌Verticillium dahliae中的生物学功能,从大丽轮枝菌落叶型菌株V592中克隆VdSRP2基因并进行生物信息学分析,利用酵母转化酶分泌系统对其信号肽活性进行测定,采用实时荧光定量PCR(real-time fluorescence quantitative PCR,qPCR)技术分析VdSRP2基因在大丽轮枝菌中的表达模式,并以V592菌株为材料获得VdSRP2基因的敲除突变体和过表达体菌株,通过表型分析和致病性测定确定VdSRP2基因的生物学功能。结果显示,VdSRP2基因编码232个氨基酸,含有5个半胱氨酸残基,N-端信号肽具有分泌活性,为真菌的典型效应蛋白;VdSRP2基因主要在大丽轮枝菌菌丝和微菌核中表达,其中经棉花根系诱导培养24 h时表达量最高;与野生型菌株V592相比,VdSRP2基因敲除导致大丽轮枝菌的产孢量和孢子萌发率显著下降,不能形成微菌核,对棉花的致病力明显减弱;但VdSRP2基因敲除不影响大丽轮枝菌的穿透能力;VdSRP2基因在本氏烟和棉花叶片瞬时表达不会诱导细胞死亡。表明VdSRP2是大丽轮枝菌微菌核形成必需...  相似文献   

12.
棉花黄萎病菌ISSR反应体系优化及其遗传多样性分析   总被引:2,自引:3,他引:2  
为给棉花黄萎病菌分子变异及遗传多样性研究提供可靠的检测方法,以棉花黄萎病菌基因组DNA为模板,采用正交优化方法对PCR体系中DNA聚合酶、引物、dNTPs、DNA模板、Mg2+及10×Buffer等重要参数进行6因素4水平优化,建立了棉花黄萎病菌ISSR-PCR优化反应体系,并从20条ISSR通用引物中筛选出多态性较好的10条引物。采用该优化反应体系和10条ISSR引物对采自陕西棉花主产区的21个棉花黄萎病菌菌株和3个参照菌株进行ISSR分析。结果显示,10条ISSR引物共扩增出87条谱带,条带分子量均在250~2 000 bp之间,平均每条引物扩增出8.7个条带,其中58条为多态性条带,占65.2%。聚类分析结果显示,在相似系数0.59处,供试菌株分为2个遗传类型。表明棉花黄萎病菌菌株间的亲缘关系与地理来源存在一定的相关性,而与其病害症状类型无相关性。  相似文献   

13.
Verticillium dahliae antagonistic endorhizosphere bacteria were selected from root tips of tomato plants grown in solarized soils. Fifty-three out of the 435 selected bacterial isolates were found to be antagonistic against V. dahliae and several other soilborne pathogens in dual cultures. Significant biocontrol activity against V. dahliae in glasshouse trials was demonstrated in three of 18 evaluated antagonistic isolates, provisionally identified as Bacillus sp. Although fluorescent pseudomonads were also isolated from root tips of tomato plants, none of the tested isolates exercised any significant antagonistic activity against V. dahliae in dual cultures. So these isolates were not tested in glasshouse trials in this study. Finally, two of the most effective bacterial isolates, designated as K-165 and 5-127, were shown to be rhizosphere colonizers, very efficient in inhibiting mycelial growth of V. dahliae in dual cultures and successfully controlling Verticillium wilt of solanaceous hosts. In glasshouse experiments, root dipping or soil drenching of eggplants with bacterial suspension of 107cfu ml–1 resulted in reduced disease severity expressed as percentage of diseased leaves (40–70%) compared to the untreated controls under high V. dahliae inoculum level (40 microsclerotia g–1 soil). In heavily Verticillium infested potato fields, experiments with potato seeds dusted with a bacterial talc formulation (108cfu g–1 formulation), showed a significant reduction in symptom development expressed as percentage of diseased potato plants and a 25% increase in yield over the untreated controls. As for their effectiveness in increasing plant height, both bacterial isolates K-165 and 5-127 produced indolebutyric, indolepyruvic and indole propionic acids. Both antagonists are considered as plant growth promoting rhizobacteria bacteria since significantly increased the height of treated plants compared with the untreated controls. Chitinolytic activity test showed that both isolates were able to produce chitinase. Testing rhizospheric and endophytic activity of the antagonists it was shown that although the bacteria are rhizosphere inhabitants they also preferentially colonize the endorhizosphere of tomatoes and eggplants. Fatty acid analysis showed that isolate K-165 could belong to Paenibacillus alvei while 5-127 to Bacillus amiloliquefaciens.  相似文献   

14.
Using differential hybridization, two DNA fragments, VDf35 and VDf90, specific to Verticillium dahliae, were isolated. These fragments contained truncated open reading frames (ORFs) homologous to the gypsy-type retrotransposon. The ORFs of VDf35 and VDf90 were pol and gag homologs, respectively. In addition, VDf90 had a pol homolog without an ORF sequence. The pol homologs in VDf35 and VDf90 were similar to each other, and these two DNA fragments had completely identical sequences. Genomic Southern analysis revealed that numerous copies of these homologs existed in V. dahliae, suggesting that V. dahliae carries a gypsy-like retroelement. Genomic Southern and polymerase chain reaction (PCR) analysis also indicated that a large number of these homologs exist in V. longisporum as well as in V. dahliae, but only a few were present in V. albo-atrum. No homolog was found in either V. nigrescens or V. tricorpus. The uneven distribution of these homologs of the retroposon-like elements among Verticillium species suggested a close genetic kinship between V. dahliae and V. longisporum. PCR primers designed from VDf35 showed species- or pathotype-specific amplification. Therefore, this sequence may be useful as a DNA marker to identify species and pathotypes of V. dahliae. This is the first report on a retrotransposon-like sequence in the genome of phytopathogenic Verticillium species.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB095264 (VDf90) and AB095265 (VDf35)  相似文献   

15.
为掌握新疆主要植棉区棉花黄萎病的发生现状及其病原菌大丽轮枝菌Verticillium dahliae的落叶型菌系分布以及遗传变异情况,于2015年对26个新疆主要植棉区棉花黄萎病的发生情况进行了随机调查,统计新疆大丽轮枝菌的培养性状,利用大丽轮枝菌落叶型特异引物D1/D2、INTD2F/INTD2R与非落叶型特异性引物ND1/ND2、INTNDF/INTNDR对新疆大丽轮枝菌菌系进行互补鉴定,并对部分菌系的遗传变异进行简单序列重复区间(inter simple sequence repeat,ISSR)分析。结果表明:2015年新疆棉花黄萎病发病田比例为54.0%,其中病情指数在10.0以上的发病田与2013年持平,而病情指数在20.0以上的严重发病田比例为10.8%,比2013年增加3.8个百分点;新疆大丽轮枝菌的培养性状以菌核型为主,比例为70.1%,菌丝型与中间型比例分别为13.4%和16.5%;新疆大丽轮枝菌落叶型菌系比例为53.2%,26株菌株的来源地全部检出落叶型菌系;聚类分析结果显示,当遗传相似系数为0.66时,新疆大丽轮枝菌落叶型与非落叶型菌系聚为2个谱系,菌系地理来源、培养性状与大丽轮枝菌的遗传分化无明显相关性。  相似文献   

16.
Complementary auxotrophic nitrate-nonutilizing (nit) mutants were used to investigate vegetative compatibility within 27 strains ofVerticillium dahliae isolated from several hosts originating from Africa, Asia, Europe and the United States. Using about 500nit mutants generated from these strains, three vegetative compatibility groups, 1, 2 and 4, were identified. Simultaneously, virulence of each strain was assessed on cultivars ofGossypium hirsutum, G. barbadense andG. arboreum, based upon Foliar Alteration Index (FAI) and Browning Index (BI) estimation. The strains in VCG1 were of both the cotton-defoliating pathotype and race 3 (on cotton) but were non pathogenic on tomato; those in VCG2 and VCG4 were of the nondefoliating pathotype and belonged to different races on cotton and on tomato. Hyaline mutants deriving from parental wild-type strain showed differences in pathogenicity but were always assigned to the parental VCG. A relationship was established between VCGs and the taxonomic position of host plants. Data fromnit pairings indicated that the sub-populations ofV. dahliae (VCGs) may not be completely isolated genetically.  相似文献   

17.
Agrobacterium tumefaciens‐mediated transformation (ATMT) was used to obtain a large number of Verticillium dahliae (Vd991) T‐DNA insertion mutants that were randomly integrated. Insertion mutants that produced significantly fewer microsclerotia were chosen for further analysis. Mutant T0065 was identified as having the Cerevisin gene interrupted by T‐DNA, and it was named cerevisin. The cerevisin protein showed a high amino acid sequence similarity with the vacuolar protease B. The mutant strain cerevisin displayed decreased production of microsclerotia and conidia, significantly reduced growth rate and reduction in virulence compared to the wild type. Moreover, the composition of secreted proteins differed between the cerevisin mutant and the wild type. Loss of function of Cerevisin decreased secretion of proteins of low molecular weight (14–25 kDa). Upon treatment with the secreted proteins of the mutant, the degree of leaf wilting decreased, indicating that Cerevisin is implicated in production of these proteins, which are putative pathogenicity factors of V. dahliae. The results suggest that Cerevisin is involved in controlling multiple processes of development and metabolism, plays an important role in vegetative growth and microsclerotia formation and affects virulence of V. dahliae.  相似文献   

18.
陕西棉花黄萎病菌致病力分化及其遗传多样性   总被引:4,自引:1,他引:4  
采用生物学培养性状、致病力测定和ISSR分子标记方法研究15个棉花黄萎病代表菌株的遗传变异.结果表明,供试菌株的生长速率、孢子产量与致病力的强弱呈正相关.致病力测定结果显示,致病力强的Ⅰ型有7个菌株,占46.67%,平均病情指数大于36.1;致病力弱的Ⅱ型有3个菌株,占20.00%,平均病情指数在21以下;致病力中等的Ⅲ型有5个菌株,占33.33%,平均病情指数在20~28之间.用4条ISSR引物对这些菌株进行PCR扩增,共得到623个条带,具多态性的有425条.聚类分析和相似系数分析结果显示,在0.55遗传相似水平下,供试菌株分为2个遗传类型,遗传类型与菌株致病力类型存在明显的相关性,与菌株地理来源也具有一定的相关性.  相似文献   

19.
Verticillium longisporum is a soil-borne fungal pathogen causing vascular wilt of Brassica crops. This study was conducted to enhance our knowledge on the host range of V. longisporum. Seven crop species (barley, oat, oilseed rape, pea, red clover, sugar beet and wheat) and five weed species (barren brome, black-grass, charlock, cleavers and scentless mayweed) all common in southern Sweden were evaluated for infection by response to V. longisporum. Oat, spring wheat, oilseed rape, scentless mayweed and charlock inoculated with V. longisporum in a greenhouse showed stunting to various degrees close to the fully ripe stage. Based on the extent of microsclerotia formation, explants were separated into four groups: for pea and wheat, <5% of the samples had formed microsclerotia; for scentless mayweed, 5–10%; for oat, 10–20%; and for charlock and oilseed rape >80%. The results suggest that plant species outside the Brassicaceae can act as reservoirs of V. longisporum inoculum. Soil inoculum densities in nine fields were monitored over a period of 12 months, which ranged from 1 to 48 cfu g−1 soil. Density of microsclerotia was lowest just after harvest, reaching its maximum six months later. No significant correlation between inoculum density in soil and disease incidence on oilseed rape plants was found. However, the data suggest that a threshold of 1 cfu g−1 soil is needed to cause disease on oilseed rape. Species identification based on microsclerotia morphology and PCR analysis showed that V. longisporum dominated in soil of seven, and V. dahliae in two of the nine fields studied.  相似文献   

20.
河南商丘地区棉花黄萎病菌分离鉴定和致病力分析   总被引:2,自引:1,他引:2  
为探讨河南商丘地区棉花黄萎病菌的致病型群体变异,对该地区棉花上分离的8株单孢菌株的菌落形态、显微结构、致病力、ITS序列、系统进化及菌体蛋白等方面进行了研究。结果表明:这些菌株均属于棉花黄萎病菌Verticilliumdahliae;系统进化树显示8株菌株并没有聚在同一进化枝上;8株黄萎菌菌株存在致病力差异,SQ4菌株致病力最强,属于落叶型,而其它致病力较弱的7个菌株属于非落叶型;不同致病力的菌株间蛋白谱带存在差异。  相似文献   

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