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1.
由大丽轮枝菌引起的黄萎病是我国棉花和茄子的主要病害之一,对棉花和茄子的生产造成巨大损失。为了研究大丽轮枝菌的群体遗传变异以及对棉花和茄子的交互致病性,本文对分离自江苏省的63个棉花黄萎菌和10个茄子黄萎菌进行了培养、遗传和致病特性分析。根据菌株在PDA培养基上生长时形成微菌核的多少来划分培养类型,结果菌核型占83.6%,成为主要的培养类型。用PCR技术检测菌株的致病类型、交配型以及是否具有无毒基因Ave1,结果落叶型菌株占86.3%,为优势种群,但是10株茄子黄萎菌100%都是非落叶型菌株;供试的所有江苏菌株交配型都是MAT1-2型,并且都没有无毒基因Ave1。选择江苏省的6个棉花黄萎菌和4个茄子黄萎菌在室内苗期接种棉花和茄子,进行交互致病性测定,结果这10个菌株都可以侵染棉花和茄子,而且来源于不同寄主作物的菌系之间致病力分化明显,表现在不同菌株对同一寄主的致病力不同,同一菌株对不同寄主的致病力也不同。研究结果为深入研究大丽轮枝菌群体遗传结构和制定黄萎病防治措施提供了理论支持。 相似文献
2.
新疆是我国最主要的棉花生产基地,产量占全国的85%左右。由大丽轮枝菌引起的黄萎病严重阻碍了新疆棉花产业的可持续发展。因此,本研究针对新疆棉区采集分离的140个菌株进行研究,结果发现不同地域菌株的生长速度和产孢量差异显著,新疆棉区以菌核型、落叶型菌株为主;与此同时,建立了一种苗期棉花黄萎病抗性快速鉴定新方法——育苗块定量接种法,并在中植棉2号和新陆早36号上测定了部分菌株的致病力,结果发现新疆棉区以强致病力、中等致病力类型的菌株占主导。此外,落叶型菌株的生长速度、产孢量及致病力均极显著高于非落叶型菌株。结果表明,新疆棉田大丽轮枝菌的组成与分化具有很强的地域性,这对品种的合理布局、抗病性筛选等具有重要意义。 相似文献
3.
2014年在石河子大学试验站和实验农场种植的红花出现了一种植株矮化,叶片发黄,枯死的病害,切开茎秆,维管束变成褐色。采用常规组织分离法对病组织茎秆进行分离、纯化获得单孢纯培养菌株;通过常规纸钵撕底沾根法对其进行致病性测定;用形态学和 rDNA-ITS 序列分析对病原菌进行鉴定。结果表明,分离菌株的菌落形态、分生孢子梗和分生孢子的形态都与大丽轮枝菌Verticillium dahliae 一致;经分子生物学鉴定,该菌的 rDNA-ITS 序列与中国棉花黄萎病菌V．dahliae 的 ITS 序列（登录号 KT803074）同源性为99％以上。故将引起新疆红花黄萎病的病原菌鉴定为大丽轮枝菌V．dahliae。 相似文献
4.
大丽轮枝菌Verticillium dahliae引起的黄萎病是一种传播迅速且为害广泛的土传维管束真菌病害。大丽轮枝菌分泌的水解酶在侵染中发挥重要作用,尤其是纤维素酶和果胶酶。大丽轮枝菌果胶裂解酶包括PL1、PL3、PL4、PL9和PL11 5个家族,其中PL1家族成员数目最多,但对该家族的致病功能还未见系统研究。本研究明确大丽轮枝菌PL1家族共包含17个成员,且多数成员在侵染早期上调表达,其中VdPL1-4和VdPL1-8在侵染早期强烈诱导表达。挑选VdPL1-4构建缺失突变体,发现VdPL1-4缺失后菌株对棉花的致病力显著下降,对复杂碳源(果胶、羧甲基纤维素、淀粉)的利用能力受限,该基因回补后菌株的毒力和碳源利用能力均得以恢复。信号肽介导的分泌活性验证表明VdPL1-4为分泌蛋白。以上研究表明,VdPL1-4是重要毒力因子,研究结果为后续系统解析果胶裂解酶在病原致病过程中发挥的作用提供了理论依据。 相似文献
5.
由大丽轮枝菌引起的棉花黄萎病是一种难以防治的土传病害,西兰花残体还对其有一定的防治作用,防效为62.82%。为解析西兰花残体对棉花黄萎病的防治机制,本研究通过土壤杆菌转化的方法构建了大丽轮枝菌的绿色荧光标记菌株,采用共聚焦显微镜观察标记菌株在添加西兰花残体营养基质中和空白对照营养基质种植的棉花体内的侵染和扩展情况。结果表明构建的标记菌株gfp-wx-1与野生菌株wx-1的生物学特性无显著性差异。同时发现大丽轮枝菌在添加西兰花残体营养基质中棉花体内扩展较慢,具体表现为:在空白营养基质种植的棉花体内,大丽轮枝菌在接种第2 d时就可侵染根尖表层及根内部,第3 d到达茎部维管束,第5 d叶片可观察到少量病原菌,第7 d第一片子叶呈现大量病原菌,后期迅速扩展,叶片出现黄萎病斑;而在营养基质中添加西兰花残体种植的棉花体内,大丽轮枝菌在接种第2 d时就可侵染根尖表层,第3 d侵染根尖内部,第5 d侵染茎部维管束,直到第7 d第一片子叶才出现少量病原菌,后期扩展慢,叶片病斑较少。本研究通过荧光定量PCR方法检测了大丽轮枝菌在两个处理棉花根部定殖情况,结果表明西兰花残体能够显著降低大丽轮枝菌在棉花根部定殖的量。该研究初步明确了西兰花残体对棉花黄萎病的防治机制,为棉花黄萎病的绿色防治提供了一条新的途径。 相似文献
6.
由大丽轮枝菌引起的棉花黄萎病是一种难以防治的土传病害,西兰花残体还对其有一定的防治作用,防效为62.82%。为解析西兰花残体对棉花黄萎病的防治机制,本研究通过土壤杆菌转化的方法构建了大丽轮枝菌的绿色荧光标记菌株,采用共聚焦显微镜观察标记菌株在添加西兰花残体营养基质中和空白对照营养基质种植的棉花体内的侵染和扩展情况。结果表明构建的标记菌株gfp-wx-1与野生菌株wx-1的生物学特性无显著性差异。同时发现大丽轮枝菌在添加西兰花残体营养基质中棉花体内扩展较慢,具体表现为:在空白营养基质种植的棉花体内,大丽轮枝菌在接种第2 d时就可侵染根尖表层及根内部,第3 d到达茎部维管束,第5 d叶片可观察到少量病原菌,第7 d第一片子叶呈现大量病原菌,后期迅速扩展,叶片出现黄萎病斑;而在营养基质中添加西兰花残体种植的棉花体内,大丽轮枝菌在接种第2 d时就可侵染根尖表层,第3 d侵染根尖内部,第5 d侵染茎部维管束,直到第7 d第一片子叶才出现少量病原菌,后期扩展慢,叶片病斑较少。本研究通过荧光定量PCR方法检测了大丽轮枝菌在两个处理棉花根部定殖情况,结果表明西兰花残体能够显著降低大丽轮枝菌在棉花根部定殖的量。该研究初步明确了西兰花残体对棉花黄萎病的防治机制,为棉花黄萎病的绿色防治提供了一条新的途径。 相似文献
7.
一个编码富含丝氨酸蛋白的基因影响大丽轮枝菌的微菌核形成、产孢及致病力 总被引：1，自引：0，他引：1
大丽轮枝菌是一种可在广泛的寄主范围内引发黄萎病的土传植物病原真菌,主要以微菌核的形式在土壤中存活多年,因此鉴定与微菌核形成及致病力相关的基因对防治该病害至关重要。本课题组从前期构建的棉花黄萎病菌T-DNA插入突变体库中筛选到一个微菌核明显减少的突变体,致病力测定结果表明,该突变体致病力明显下降。以棉花黄萎病菌野生型菌株V592的基因组DNA为模板,从棉花黄萎病菌中克隆到被T-DNA插入突变的基因(VdSRP1)编码区全长为415 bp,包含一个外显子,编码一个富含丝氨酸蛋白,与任何已知的注释基因没有显著的序列相似性。为了明确VdSRP1基因在大丽轮枝菌中的功能,利用同源重组的原理及农杆菌介导的遗传转化方法获得了VdSRP1基因的2个敲除体菌株,与棉花黄萎病菌野生型菌株V592相比,VdSRP1基因敲除突变体的微菌核形成明显减少,产孢量及孢子萌发率下降,对棉花的毒力也显著下降。对野生型菌株V592及VdSRP1敲除突变体的转录分析表明,VdSRP1调控一系列与微菌核形成、孢子形成及致病力相关基因的表达。这些结果表明,VdSRP1基因影响大丽轮枝菌的致病力、微菌核形成、产孢及孢子萌发。 相似文献
8.
大丽轮枝菌是一种可在广泛的寄主范围内引发黄萎病的土传植物病原真菌,主要以微菌核的形式在土壤中存活多年,因此鉴定与微菌核形成及致病力相关的基因对防治该病害至关重要。本课题组从前期构建的棉花黄萎病菌T-DNA插入突变体库中筛选到一个微菌核明显减少的突变体,致病力测定结果表明,该突变体致病力明显下降。以棉花黄萎病菌野生型菌株V592的基因组DNA为模板,从棉花黄萎病菌中克隆到被T-DNA插入突变的基因(VdSRP1)编码区全长为415 bp,包含一个外显子,编码一个富含丝氨酸蛋白,与任何已知的注释基因没有显著的序列相似性。为了明确VdSRP1基因在大丽轮枝菌中的功能,利用同源重组的原理及农杆菌介导的遗传转化方法获得了VdSRP1基因的2个敲除体菌株,与棉花黄萎病菌野生型菌株V592相比,VdSRP1基因敲除突变体的微菌核形成明显减少,产孢量及孢子萌发率下降,对棉花的毒力也显著下降。对野生型菌株V592及VdSRP1敲除突变体的转录分析表明,VdSRP1调控一系列与微菌核形成、孢子形成及致病力相关基因的表达。这些结果表明,VdSRP1基因影响大丽轮枝菌的致病力、微菌核形成、产孢及孢子萌发。 相似文献
9.
Nit突变体营养亲和性技术在棉花黄萎病菌生理型测定中的应用 总被引：6，自引：0，他引：6
大丽轮枝菌(Verticullrum clahliue Kleb.)是世界性的土传病原菌,严重为害棉花等多种作物。姚耀文等(1982)根据我国棉花黄萎病菌在3大棉种上的致病性反应,将致病力最强的陕西泾阳菌系定为生理型I;致病力最弱的新疆和田菌系为生理型Ⅱ:致病力中等的河南安阳茵系为生理型Ⅲ。 相似文献
10.
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
11.
Classification of 32 Verticillium dahliae isolates originating from 19 plant species in eight different botanical families to races and determination of host range pathogenicity were carried out. The physiological races of isolates were identified using the two differential tomato cultivars ??Belladonna?? (susceptible to both races 1 and 2 of V. dahliae) and ??Ace 55VF?? (resistant to race 1, susceptible to race 2 of V. dahliae). Among these isolates, 14 were race 2 (43.8%), 12 race 1 (37.5%) and six nonpathogenic (18.7%) on tomato. The host range pathogenicity of isolates was determined using four differential hosts (eggplant, turnip, tomato (Ve ? ) and sweet pepper). Among isolates, five were pathogenic to both eggplant and turnip (15.6%), 21 to eggplant, turnip and tomato (65.6%), five to eggplant, turnip, tomato and sweet pepper (15.6%) and one was pathogenic to eggplant, turnip and sweet pepper (3.2%). The pathogenicity of isolates on the aforementioned five hosts was investigated on the basis of external symptoms and by calculating the relative areas under disease progress curves (relative AUDPC). Results showed that eggplant was the most susceptible, followed by turnip and tomato cv. Belladonna, while sweet pepper and tomato cv. Ace 55VF were less susceptible to all the isolates used. The pathogenicity of isolates varied from highly to mildly virulent on eggplant and turnip while on Belladonna, Ace 55VF and sweet pepper it varied from highly virulent to nonpathogenic. Belladonna exhibited a similar level of susceptibility to races 1 and 2 of V. dahliae, but was more susceptible than Ace 55VF to race 2. Interestingly, the isolates originating from eggplant were clearly more virulent than those originating from tomato and black nightshade on all solanaceous plants tested. 相似文献
12.
苎麻疫霉寄生专化性研究 总被引：3，自引：1，他引：3
从棉铃、棉苗、构树、苧麻等植物上分离的苧麻疫霉(Phytophthora boehmeriae)菌株对上述各寄主植物进行交互接种测定致病性结果表明,分离自上述寄主植物的苧麻疫霉菌株对上述3种植物均能致病,表明不同寄主来源的苧麻疫霉菌株间致病性不存在对寄主植物的寄主专化性,但对各寄主植物的致病力有一定差异。此外,分离自棉苗和棉铃的苧麻疫霉各菌株对棉苗和棉铃具有相似的致病力;分离自构树的菌株多次接种棉苗后致病力逐渐增强,初步认为构树是棉疫病菌的野生寄主。 相似文献
13.
Host Range Specificity in Verticillium dahliae 总被引：1，自引：0，他引：1
ABSTRACT Verticillium dahliae isolates from artichoke, bell pepper, cabbage, cauliflower, chili pepper, cotton, eggplant, lettuce, mint, potato, strawberry, tomato, and watermelon and V. albo-atrum from alfalfa were evaluated for their pathogenicity on all 14 hosts. One-month-old seedlings were inoculated with a spore suspension of about 10(7) conidia per ml using a root-dip technique and incubated in the greenhouse. Disease incidence and severity, plant height, and root and shoot dry weights were recorded 6 weeks after inoculation. Bell pepper, cabbage, cauliflower, cotton, eggplant, and mint isolates exhibited host specificity and differential pathogenicity on other hosts, whereas isolates from artichoke, lettuce, potato, strawberry, tomato, and watermelon did not. Bell pepper was resistant to all Verticillium isolates except isolates from bell pepper and eggplant. Thus, host specificity exists in some isolates of V. dahliae. The same isolates were characterized for vegetative compatibility groups (VCGs) through complementation of nitrate nonutilizing (nit) mutants. Cabbage and cauliflower isolates did not produce nit mutants. The isolate from cotton belonged to VCG 1; isolates from bell pepper, eggplant, potato, and tomato, to VCG 4; and the remaining isolates, to VCG 2. These isolates were also analyzed using the random amplified polymorphic DNA (RAPD) method. Forty random primers were screened, and eighteen of them amplified DNA from Verticillium. Based on RAPD banding patterns, cabbage and cauliflower isolates formed a unique group, distinct from other V. dahliae and V. albo-atrum groups. Minor genetic variations were observed among V. dahliae isolates from other hosts, regardless of whether they were host specific or not. There was no correlation among pathogenicity, VCGs, and RAPD banding patterns. Even though the isolates belonged to different VCGs, they shared similar RAPD profiles. These results suggest that management of Verticillium wilt in some crops through crop rotation is a distinct possibility. 相似文献
14.
新疆加工型辣椒细菌性斑点病的发生和病原鉴定 总被引：4，自引：0，他引：4
新疆加工型辣椒主要产区巴音郭楞蒙古自治州发生了一种严重危害辣椒的细菌性病害。从发病辣椒叶片中分离细菌,通过烟草过敏性反应、马铃薯软腐试验和接种辣椒等致病性测定,确定了13个致病菌株,各菌株之间致病力无明显差异。通过菌体形态、培养性状观察、生理生化反应、寄主范围测定,结合16S rDNA和rpoD基因扩增、序列测定和系统发育分析,将病原菌鉴定为丁香假单胞菌丁香致病变种(Pseudomonas syringae pv. syringae)。病原菌人工接种还能侵染番茄、茄子、马铃薯及黄瓜、四季豆、白菜、萝卜、芹菜等植物。P. syringae pv. syringae引起加工型辣椒细菌性斑点病在国内属首次报道。 相似文献
15.
棉花黄萎病菌毒素对棉花作用机制的初步探讨 总被引：9，自引：0，他引：9
用棉花黄萎病菌(Verticillium dahliae Kled.) T-9、SS4、VD-8、VD-5等4个菌株的毒素(VD-toxins)处理棉苗,发现各菌株的毒素都能使棉苗体内脱落酸(ABA)的含量增加,其中以T-9菌株毒素的影响最明显。各菌株毒素均可增加棉苗叶片的气孔抗性和提高叶片细胞的离子渗透率3-4倍。毒素对棉苗叶片的游离原生质体具有30-50%的损伤率。 相似文献
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Nadia Korolev Encarnación Pérez-Artés Jesús Mercado-Blanco José Bejarano-Alcázar Dolores Rodríguez-Jurado Rafael M. Jiménez-Díaz Talma Katan Jaacov Katan 《European journal of plant pathology / European Foundation for Plant Pathology》2008,122(4):603-617
Verticillium dahliae isolates recovered from a new focus of severe Verticillium wilt of cotton in the northeast of Israel were tested for vegetative
compatibility using nitrate non-utilizing (nit) mutants and identified as VCG1, which is a new record in Israel. Other cotton isolates of V. dahliae from the northern and southern parts of the country were assigned to VCG2B and VCG4B, respectively. VCG1 isolates induced
severe leaf symptoms, stunting and defoliation of cotton cv. Acala SJ-2, and thus were characterized as the cotton-defoliating
(D) pathotype, whereas isolates of VCG2B and VCG4B were confirmed as the earlier described defoliating-like (DL) and non-defoliating
(ND) pathotypes, respectively. This is the first record of the D-pathotype in Israel. The host range of representative isolates
of each VCG-associated pathotype was investigated using a number of cultivated plants. Overall, the D isolates were more virulent
than DL isolates on all tested host plants, but the order of hosts (from highly susceptible to resistant) was the same: okra
(Hibiscus esculentus local cultivar), cotton (Gossypium hirsutum cv. Acala SJ2), watermelon (Citrullus lanatus cv. Crimson Sweet), safflower (Carthamus tinctorius cv. PI 251264), sunflower (Helianthus annuum cv. 2053), eggplant (Solanum melongena cv. Black Beauty), and tomato (Lycopersicon esculentum cv. Rehovot 13). The pattern of virulence of ND isolates differed from that of D and DL isolates, so that the former were
highly virulent on eggplant but mildly virulent on cotton. Tomato was resistant to all cotton V. dahliae isolates tested. RAPD and specific PCR assays confirmed that the D isolates from Israel were similar to those originating
from other countries. 相似文献
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Ioannis A. Papaioannou Eleftherios K. Ligoxigakis Demetrios J. Vakalounakis Emmanouil A. Markakis Milton A. Typas 《European journal of plant pathology / European Foundation for Plant Pathology》2013,136(3):577-596
A population of 84?V. dahliae isolates mainly originating from Crete, Greece, was characterized in terms of pathogenicity and virulence on different hosts, in parallel with morphological/physiological characterization, vegetative compatibility grouping and mating type determination. Tomato race 2 was found to have supplanted race 1 and was more virulent on a tomato-susceptible cultivar than race 1. Using a differential host classification system which tests pathogenicity to tomato, eggplant, sweet pepper and turnip, 59 isolates were assigned to tomato, 19 to eggplant, one to sweet pepper and five to tomato-sweet pepper pathogenicity groups. All isolates from Crete fell into VCG subgroups 2A, 2B and 4B, while a remarkably high incidence of bridging isolates (compatible with two or more VCGs) was recorded. The tomato-sweet pepper pathogenicity group was morphologically quite distinct from the others, while conidial length and pigment intensity were discriminatory parameters among VCGs 2A, 2B and 4B. PCR-based molecular marker Tr1/Tr2 was reliable in race prediction among tomato-pathogenic isolates, except for members of VCG 4B, while the application of markers Tm5/Tm7 and 35-1/35-2 was highly successful for tomato-pathogenic isolates. E10 marker was related to VCG 2B, rather than to pathogenicity groups. A single nucleotide polymorphism in the ITS2 region, and two novel molecular markers, M1 and M2, proved useful for the fast and accurate determination of major VCGs 2A, 2B and 4B, and can be used for high-throughput population analyses in future studies. The mating type was unrelated to VCG classification and probably does not control heterokaryon incompatibility in V. dahliae. 相似文献
20.
A weed survey conducted in 2004 and 2005 in Aydin province of Turkey showed that Solanum nigrum, Xanthium strumarium, Amaranthus retroflexus, Portulaca oleracea, Sonchus oleraceus and Datura stramonium were the most prevalent weeds in the cotton fields exhibiting Verticillium wilt. Verticillium dahliae Kleb. was recovered from A. retroflexus and X. strumarium in those cotton fields. This is the first report of V. dahliae occurring naturally in A. retroflexus in Turkey. Pathogenicity tests on cotton and weeds showed that the virulence of V. dahliae isolates from weeds was higher on cotton plants than on weeds, with the disease severity ranging from 31.7% to 98.0%. Disease
severity of V. dahliae isolates was 54.7–93.9% on eggplant, 23.7–51.6% on cucumber and 11.0–16.4% on tomato, whereas it did not cause any disease
symptoms, or only low levels, on pepper and bell pepper. Two vegetative compatibility groups (VCGs) were identified among
seven tested weed isolates: VCG2A (two isolates) and VCG2B (three isolates) using international reference strains. 相似文献