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1.
新疆北部棉区黄萎病菌种群致病性分化及变异 总被引：6，自引：2，他引：4
采用特异性引物PCR检测技术和鉴别寄主法对新疆北部棉区41个棉花黄萎病菌菌系进行检测,以期明确黄萎病菌种群致病性分化及变异。特异性引物(ND1/ND2和D1/D2)PCR检测结果显示,供试菌系中落叶型菌系16个,占39.0%;非落叶型菌系24个,占58.5%;1个菌系未能检测出致病类型。鉴别寄主法测定结果显示:41个供试菌株中致病性强、中、弱的菌系分别为21、12和8个,分别占51.2%、29.3%和19.5%。落叶型菌系的致病力明显高于非落叶型,平均病情指数分别为39.4和25.8。研究表明,无论落叶型菌系和非落叶型菌系,接种后都可产生落叶症状,但落叶的程度有明显差别,其落叶症状的轻重不仅与菌系的致病类型有关,还与品种的抗病性密切相关。 相似文献
2.
棉花黄萎病菌致病型的AFLP分析 总被引：16，自引：0，他引：16
选用41个棉花黄萎病菌(Verticillium dahliae)代表菌系,在温室条件下,对4个棉花品种鄂荆1号(感)、中棉所12(耐)、文-5(抗)和唐棉2号(抗)进行致病性测定,结果可将供试菌系分为落叶型与非落叶型2类。选取8对AFLP引物PCR扩增的结果中,统计带型稳定、清晰且有多态性的条带,共169条作系统聚类分析,将上述菌系分为2大类,第一类为非落叶型菌系,包括10个非落叶型菌系和1个过渡菌系;第二类为30个落叶型菌系。根据聚类分析建立树状图,发现菌系与地理来源存在一定的相关性,而依据菌系致病力强弱分类则相关关系不大。选用25对EcoRⅠ和MseⅠ引物组合,对供试的41个V.dahliae进行AFLP扩增,筛选到2对引物E64(GACTGCGTACCAATTCGAC)、M53(GATGAGTCCTGAGTAACCG)和E49(GACTGCGTACCAATTCCAG)、M65(GAT-GAGTCCTGAGTAAGAG),能分别扩增出433bp和110bp2条仅为V.dahliae非落叶型菌系独有的特异片段,可将落叶型与非落叶型菌系分开,这2条特异片段被命名为EM433和EM110。 相似文献
3.
新疆棉花黄萎病的发生现状及其病原菌的分子鉴定与ISSR分析 总被引：1，自引：0，他引：1
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
4.
北方棉区棉花黄萎病菌RAPD分析 总被引：13，自引：0，他引：13
以１４个黄萎病菌代表菌系为对照，对来自我国北方棉区的３４个棉花黄萎病菌菌系进行ＲＡＰＤ分析。选用对所有供试菌系都有扩增条带的１４个引物，取其结果中稳定性和多态性均好的６５条谱带作类平均法系统聚类分析，建立树状图。将上述４８个菌系分为４大类，结果表明北方棉区河北、河南、山东的部分棉田存在黄萎病菌落叶型菌系的危害，而且８５．７％的落叶型菌系与对照的美国落叶型菌系Ｔ９、Ｖ４４的亲缘关系比与对照的江苏落叶型菌系ＶＢ、Ｖ９９１更接近。 相似文献
5.
北方棉区棉花黄萎病菌落叶型菌系鉴定 总被引：19，自引：1，他引：18
采用RAPD扩增与温室致病性测定2种方法,以黄萎病菌Verticillium dahliae的5个落叶型、7个非落叶型菌系和V.albo-atrum的2个菌系为对照,对采自北方棉区6省(自治区)的34个棉花黄萎病菌菌系进行致病型鉴定。94.1%的菌系在2种鉴定结果中表现一致,与对照菌系相比较,确定其中26个菌系为落叶型菌系,6个菌系为非落叶型菌系。从而证实了黄萎病菌落叶型菌系在北方棉区河北、河南、山东3省的存在,并发现所确定的26个北方落叶型菌系中的22个与来自美国的对照落叶型菌系T9、V44的关系比与来自江苏的对照落叶型菌系VB、V991的关系更近。本实验还初步筛选到2条用于鉴别V.dahliae落叶型与非落叶型菌系的RAPD特异条带OPB-19966和OPM-201691,将它们用于对34个北方菌系的RAPD扩增鉴定,则与温室致病性测定结果的一致性分别为88.2%和94.1%,证明这2条特异条带在鉴定棉花黄萎病菌落叶型菌系上具有一定的实用价值,并为进一步制作特异探针以形成一套简便、准确、规范化的鉴定技术奠定了基础。 相似文献
6.
为建立可同时快速检测引起向日葵黄萎病害的2种检疫性病原菌大丽轮枝菌Verticillium dahliae Kleb.和黑白轮枝菌V.albo-atrum Reinke et Berthold的方法,根据2种病原菌的β-tubulin基因分别设计特异性DPO引物,建立多重DPO-PCR检测方法,并对其特异性和灵敏度进行评价。结果表明,所设计的DPO引物特异性强,仅大丽轮枝菌和黑白轮枝菌可分别扩增出225 bp与151 bp的特异性条带,其它向日葵病害的7种病原菌及阴性对照均无目的条带;反应体系中引物终浓度为0.2μmol/L、退火温度为60℃时,30个扩增循环的检测灵敏度均可达0.05 ng菌丝DNA量;在45~65℃退火温度范围内均可高效扩增靶基因片段,表明该方法退火温度范围宽。所建立的检测方法能够准确、高效地检测引起向日葵黄萎病的大丽轮枝菌和黑白轮枝菌,可用于向日葵种子带菌筛查及田间病害诊断检测。 相似文献
7.
不同植棉省区落叶型黄萎病菌的培养特性及致病力比较 总被引：2，自引：1，他引：1
近年来落叶型症状的棉花黄萎病对棉花生产的危害越来越大,为明确落叶型棉花黄萎病菌的分化特点,采用特异性分子标记方法,对我国10个主要植棉省区30个落叶型棉花黄萎病菌的培养特性和致病力进行了比较。落叶型黄萎病菌系在我国主要植棉省区已普遍存在,该菌系以菌核型为主,其微菌核呈放射状或环状。63.3%菌系的菌丝发达致密,其余菌系的菌丝较为疏松。落叶型黄萎病菌系之间的产孢量和致病力差异较大,产孢量变幅为3.7×107～18.8×107个孢子/mL;71.9%为强致病力菌系,25.0%为中等致病力。研究表明,不同植棉省区间落叶型黄萎病菌系的产孢量和致病力具有一定差异。 相似文献
8.
棉花黄萎病病原菌大丽轮枝菌的快速分子检测 总被引：5，自引：0，他引：5
大丽轮枝菌Verticillium dahliae是引起棉花黄萎病的土传病害病原真菌。快速及时地检测出大丽轮枝菌,对棉花黄萎病的早期预警及后期防治具有重要意义。采用聚合酶链式反应(polymerase chain reaction,PCR)检测技术对已报道大丽轮枝菌的检测引物进行验证、筛选和改进,获得1对改进的特异性PCR引物VDS-F/VDS-R。在优化的反应体系与扩增条件下,能特异性地从大丽轮枝菌基因组扩增出l条约520 bp的产物条带,检测灵敏度达到10~(-2)ng/μL;利用该引物可特异性地从含有大丽轮枝菌的土壤及棉花植株组织中检测出病原菌;采用巢氏PCR法对人工病土的检测灵敏度达到了10个孢子/g土。表明本引物的PCR检测体系可用于棉花黄萎病的早期快速检测。 相似文献
9.
采用Nit硝酸还原酶缺陷型突变体技术，共得到棉花黄萎病菌Nit突变体301个，其中A型155个，B型97个，C型29个。共检测到落叶型棉花黄萎病菌11株，除以前报道的6株外，发现了5个新菌株。针刺接种后，在棉株上均引起典型的落叶型症状。通过聚丙烯酰受凝胶电泳比较落叶型和非落叶型的8个代表菌株的蛋白质电泳图谱的差异，发现在浓度为7.5%的凝胶中，落叶型菌株有两条特异性的蛋白，其迁移率分别为Rf5=0.135,Rf9=0.405。用非落叶型菌株DF4、落叶型菌株SY12菌体的可溶性蛋白制备获得两个抗血清。用间接ELISA法检测，即能区分棉花黄萎病菌的立枯、枯萎、炭疽病菌，落叶型和非落叶型黄萎病菌均呈阳性反应，但落叶型黄萎病菌的OD值高于非落叶型黄萎病菌。 相似文献
10.
新疆主要棉区棉花黄萎病发生概况 总被引：11，自引：0，他引：11
为了解目前新疆棉花黄萎病的发生现状,对新疆主要植棉地区棉花黄萎病发生程度、落叶型菌系分布以及代表棉区主栽品种黄萎病抗性类型、棉花黄萎病发生规律进行了调查研究。结果表明:在调查的新疆棉田中棉花黄萎病发病田占58.2%,其中病情指数达5.0以上的棉田占28.1%;新疆棉花黄萎病菌系中37.3%为落叶型菌系;阿克苏棉区主栽的棉花品种(系)‘中棉49号’、‘中棉414号’、‘2905’棉花黄萎病抗性表现为耐病,生产上缺乏抗病品种。阿克苏棉区棉花黄萎病发生始期较往年趋早,发病程度趋于严重,6月下旬至7月下旬为棉黄萎病快速发展期,后期由于高温抑制作用,发病趋缓。 相似文献
11.
Vegetative Compatibility Groupings of Verticillium dahliae from Cotton in Mainland China 总被引：2，自引：0，他引：2
Xia Zhengjun P.N. Achar Gu Benkang 《European journal of plant pathology / European Foundation for Plant Pathology》1998,104(9):871-876
One hundred and fourteen isolates of Verticillium dahliae obtained from cotton and eggplant in mainland China were successfully assigned to two vegetative compatibility groups (VCGs) except for one self-incompatible isolate. Eleven isolates were strongly compatible with T9, the tester strain of the cotton defoliating pathotype, forming a linear growth of wild type with abundant microsclerotia and dense mycelia between compatible nitrate-nonutilizing mutants. The remaining 102 isolates were grouped into the non-defoliating VCG2, although the strength of the reaction varied; some isolates were strongly compatible with the tester strain while others were only slightly compatible. All VCG1 isolates including T9 showed the same defoliating symptom in greenhouse inoculation tests. This study confirmed the presence of the defoliating pathotype (VCG1) of V. dahliae in mainland China. 相似文献
12.
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
13.
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：3，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
14.
Jiménez-Díaz RM Olivares-García C Landa BB del Mar Jiménez-Gasco M Navas-Cortés JA 《Phytopathology》2011,101(3):304-315
Severity of Verticillium wilt in olive trees in Andalusia, southern Spain is associated with the spread of a highly virulent, defoliating (D) Verticillium dahliae pathotype of vegetative compatibility group 1A (VCG1A) but the extent of this spread and the diversity of the pathogen population have never been documented. VCG typing of 637 V. dahliae isolates from 433 trees in 65 orchards from five olive-growing provinces in Andalusia indicated that 78.1% were of VCG1A, 19.8% of VCG2A, 0.6% of VCG2B, 1.4% of VCG4B, and one isolate was heterokaryon self-incompatible. A single VCG prevailed among isolates within most orchards but two and three VCGs were identified in 12 and 3 orchards, respectively, with VCG1A+VCG2A occurring in 10 orchards. VCG1A was the predominant VCG in the three most important olive-growing provinces, and was almost as prevalent as VCG2A in another one. Molecular pathotyping of the 637 isolates using specific polymerase chain reaction assays indicated that VCG1A isolates were of the D pathotype whereas isolates of VCG2A, -2B, and -4B were of the less virulent nondefoliating (ND) pathotype. The pathotype of isolates correlated with the disease syndrome affecting sampled trees. Only three (seq1, seq2, and seq4) of the seven known sequences of the V. dahliae-specific 539- or 523-bp amplicon were identified among the 637 isolates. Distribution and prevalence of VCGs and seq sequences among orchards indicated that genetic diversity within olive V. dahliae in Andalusia is higher in provinces where VCG1A is not prevalent. Log-linear analysis revealed that irrigation management, source of irrigation water, source of planting stock, and cropping history of soil were significantly associated with the prevalence of VCG1A compared with that of VCG2A. Multivariate analyses using a selected set of agricultural factors as variables allowed development of a discriminant model for predicting the occurrence of D and ND pathotypes in the area of the study. Blind tests using this model correctly indentified the V. dahliae pathotype occurring in an orchard. The widespread occurrence and high prevalence of VCG1A/D pathotype in Andalusia have strong implications for the management of the disease. 相似文献
15.
Toshiyuki USAMI Mafumi ABIKO Masahiro SHISHIDO Yoshimiki AMEMIYA 《Journal of General Plant Pathology》2002,68(2):134-140
Verticillium dahliae Klebahn is the causal agent of tomato wilt disease. Isolates of V. dahliae can be classified based on pathogenicity to tomato, but the pathotypes are indistinguishable in morphology. We designed PCR
primers for specific detection of isolates pathogenic to tomato (tomato pathotype) from the sequences of a pathotype-specific
gene, vdt1. With the primer pair Tg5/Tc3, a PCR product (approximately 3.2 kb) specific to tomato pathotype was amplified from the genomic
DNA of isolates. Using the primer pair, a tomato pathotype isolate was specifically detected from hypocotyls of inoculated
tomato and eggplant. On the other hand, no amplification was observed from non-tomato pathotype isolates of V. dahliae, some other wilt pathogens of tomato and a healthy host plant. Therefore, the primer pair can be useful for pathotype-specific
detection of V. dahliae as well as for diagnosis of wilt disease of tomato plant.
Received 7 September 2001/ Accepted in revised form 3 December 2001 相似文献
16.
Navas-Cortés JA Landa BB Mercado-Blanco J Trapero-Casas JL Rodríguez-Jurado D Jiménez-Díaz RM 《Phytopathology》2008,98(2):167-180
The development of Verticillium wilt epidemics in olive cv. Arbequina was studied from November 1999 to May 2003 in a drip-irrigated, nontillage orchard established in a soil without a history of the disease at Córdoba, southern Spain. Disease incidence measured at 1-month-intervals increased from 0.2 to 7.8% during this period. Verticillium dahliae infecting the trees was characterized as defoliating (D) or nondefoliating (ND) pathotypes by a specific, multiplex-polymerase chain reaction (PCR) assay. Of the symptomatic trees, 87.2 and 12.8% were infected by the D or ND pathotypes, respectively. Dynamics of disease incidence were described by a generalized logistic model with a multiple sigmoid pattern. In the fitted model, the infection rate was highest in the winter to spring period and decreased to minimum values in the summer to fall period. Binary data of disease incidence was analyzed for point pattern and spatial correlation, either directly or after parsing them in contiguous quadrats. Overall, ordinary runs analysis indicated a departure from randomness of disease within rows. The binomial index of dispersion, interclass correlation, and Taylor's power law for various quadrat sizes suggested aggregation of diseased trees within the quadrat sizes tested. Spatial analysis by distance indices showed a nonrandom arrangement of quadrats containing infected trees. Spatial pattern was characterized by the occurrence of several clusters of infected trees. Increasing clustering over time was generally suggested by stronger values of clustering index over time and by the increase in the size of patch clusters. Significant spatial association was found in the clustering of diseased trees over time across cropping seasons; however, clustering was significant only for infections by D V. dahliae, indicating that infections by the D pathotype were aggregated around initial infections. The number and size of clusters of D V. dahliae-infected trees increased over time. Microsatellite-primed PCR assays of a representative number of V. dahliae isolates from diseased trees indicated that the majority of infecting D isolates shared the fingerprinting profile with D V. dahliae isolated from soil of a naturally infested cotton field in close proximity to the orchard, suggesting that short distance dispersal of the pathogen from this soil to the olive orchard may have occurred. 相似文献