棉花黄萎病抗性遗传和分子生物学研究进展

介绍了棉花黄萎病菌的致病机制 ,棉花黄萎病抗病机制及其遗传以及棉花黄萎病的鉴定方法 ,并对分子生物学在棉花黄萎病研究中的应用状况进展进行了综述。     

棉花黄萎病菌与抗黄萎病遗传育种研究进展

简要综述了棉花黄萎病菌及抗黄萎病遗传育种的研究进展。研究表明 ,各地的棉花黄萎病菌均存在致病力的分化 ,其致病机理是病菌侵入棉花后菌丝及孢子在导管内大量繁殖 ,同时刺激邻近的薄壁细胞产生胶状物质及侵填体而堵塞导管 ,使水分和养分运输发生困难 ,更重要的是病菌在棉株体内产生的糖蛋白毒素作用的结果。棉花抗黄萎病的遗传方式争论较大 ,但一般在温室由单一菌系接种鉴定时棉花黄萎病抗性表现为单基因遗传 ,而在田间病圃或用多菌系混合鉴定时 ,棉花黄萎病抗性表现为多基因遗传。由于陆地棉内缺乏高抗黄萎病资源 ,给棉花抗黄萎病育种带来一定困难 ,但 90年代以来 ,已育成 86 - 6、川 73 7、川 2 80 2、豫棉 1 9号、豫棉 2 1号等一些抗黄萎病的新品种。上述抗黄萎病品种在棉花黄萎病综合防治中起了重要作用     

棉花主要病害研究概要

以棉花枯、黄萎病为主,就我国近50年来,在棉花病害研究方面取得的进展做了较详细的总结."全国棉花枯、黄萎病协作组"的成立,极大地促进了棉花病害研究工作的进展,突出表现在棉花枯、黄萎病菌生理型(小种)的鉴定,棉花的抗病机理和抗病鉴定方法研究,棉花种质资源材料和新品种系的抗病性鉴定,落叶型黄萎病的研究,抗病育种及种衣剂的研制等方面.并认为应加强棉花抗枯、黄萎病遗传,黄萎病菌致病力分化,以及抗虫棉病害发生特点、机理及抗病育种等方面的研究工作.     

我国棉花黄萎病研究进展

本文主要综述了我国７０年代以来，在棉花黄萎病发生与防治、病害消长动态规律、病原菌“种”和致病类型分化、落叶型黄萎病菌的确定及病菌致病机理、抗病性机制等方面的应用基础研究取得的新进展。并提出了加强对黄萎病防治研究与继续深入探讨的方向。     

棉花黄萎病菌致病力分化与寄主抗病性遗传研究进展

棉花黄萎病是世界各主要产棉国家和地区普遍发生和危害最重的病害之一，本文综述了棉花黄萎病菌致病力分化与寄主抗病性遗传研究进展。病菌种内存在着明显的致病力分化。寄主抗病性的遗传在种间和种内材料间亦存有差异。为了定向而有效地培育抗病品种、合理进行品种布局，对此类问题仍需进一步研究和探讨。     

我国棉花黄萎病菌基于SSR的遗传多样性分析

为了对棉花黄萎病菌群体的遗传多样性进行研究,以棉花黄萎病菌基因组DNA为模板,用双因素和单因素水平优化的方法对SSR反应体系的重要参数进行摸索和优化,建立了最适反应体系.从300对引物中筛选出13对多态性较高的引物,并对来自我国12个省96个县(市)的170个棉花黄萎病菌株进行SSR分析.SSR图谱的聚类分析结果将供试...     

棉花抗黄萎病机制研究进展

棉花黄萎病是一种土传真菌维管束病害,严重影响棉花产量和纤维品质。常规的防治手段可以局部控制但不能有效防治,采用传统的抗病育种策略培育抗病品种成效缓慢,因此对其防治一直是棉花生产上的难题。目前越来越多的研究集中在棉花对黄萎病的抗病机制方面。本文结合其他植物抗病研究进展从抗病基因介导的信号路径、乙烯在棉花与黄萎病菌互作中的作用、棉花对黄萎病菌的生理生化抗性以及棉花组织结构与黄萎病菌的抗性等4个方面总结棉花抗黄萎病的机制,以望对棉花抗病分子育种提供借鉴。     

河北省棉花黄萎病菌致病性与ISSR遗传分化

 从河北省17个主要植棉县采集棉花黄萎病株,分离获得52个黄萎病菌单孢菌系,对其培养特性、致病性和ISSR(Inter-simple sequence repeat)遗传分化进行了研究。菌系培养性状研究表明,在采集的52个菌系中,菌核型菌系最多,其次是菌丝型,最少的是中间型,3种类型菌系分别占总菌系的51.9%,38.5%和9.6%。利用7个抗、感不同的鉴别寄主在光、温、湿可控的生长室鉴定了病菌的致病性,供试菌系可分为致病力强、中、弱3种类型(Ⅰ型、Ⅱ型和Ⅲ型),分别占供试菌系的51.9%,21.2%和26.9%。在供试菌系中存在比落叶型菌系致病力还要强的非落叶型菌系。基于病情指数的聚类分析结果表明,河北省棉花黄萎病菌系存在明显致病力分化,但与地理来源无关。菌核型菌系和中间型菌系多表现为强致病力或中等致病力,而菌丝型菌系的致病力变化较大。在136个ISSR标记中,80个属于多态性标记,多态性位点百分率达58.8%。基于ISSR的聚类分析结果表明,河北省棉花黄萎病菌的遗传分化较小,并且遗传分化与地理来源相关。     

荧光标记基因转化棉花黄萎病菌及标记菌系选育

为了研究黄萎病菌的侵染机制,通过农杆菌介导的转化方法,将绿色荧光蛋白基因sGFP导入落叶型黄萎病菌VD07038,将红色荧光蛋白基因mCherryRFP导入非落叶型黄萎病菌Bp2中,分别获得了具有绿色、红色荧光信号的阳性转化子。经过分子验证和连续继代培养,证明了这些转化子具有遗传稳定的对潮霉素的抗性。通过对转化子的菌落形态、生长速度和致病力进行检测,发现大部分转化子与野生型基本一致,少量转化子发生变异,其中转化子Bp2R-30不能产生微菌核,致病力显著下降。利用荧光显微镜观察了转化子VD07038G-10在感病棉花品种苏棉22幼苗根部的侵染情况。结果表明,在接菌12 h后VD07038G-10的孢子可吸附在根表面;接种7~9 d后,菌丝入侵到棉花根部的维管组织。本研究获得的荧光蛋白标记的棉花黄萎病菌VD07038G-10可用于实时观测黄萎病菌侵染棉花根系的过程,并且可以定量鉴定不同棉花品种对该菌系的抗性,为棉花黄萎病菌抗性鉴定提供一种新方法。     

第二次国际黄萎病座谈会

1976年11月在美国加利福尼亚大学举行第二次国际黄萎病座谈会。从第一次国际黄萎病座谈会时起,就指出黄萎病菌变异性很大。正如这次座谈会组织者所指出的那样:黄萎病菌能够侵染新的作物,科学家们就黄萎病菌的致病机制和变异性阐明了新的观点,同时,对防治黄萎病问题提出了新的设想。针对病原生理学和生物化学,轮枝菌变异性遗传,抗性来源,流行病学和防治措施等问题发表了53篇专题报告,这些报告对上述问题都作了重点论述。黄萎病是一种普遍性的病害,在世界上对棉花危害性最大。美国加利福尼亚州和得     

棉花黄萎病研究进展

由大丽轮枝菌引起的棉花黄萎病已成为当前我国棉花可持续生产的主要障碍之一。该病害为土传种传维管束病害,具有病原菌寄主范围广,防治困难的特点。本文综述了近年来国内外有关其病原菌的致病力分化、全基因组测序与致病机制、微菌核形成与萌发机制,寄主的抗病分子机制以及病害防治措施等方面的最新研究进展。     

连作对棉田土壤枯、黄萎病菌数量及细菌群落的影响

采用选择培养法测定新疆库尔勒不同连作年限棉田土壤枯、黄萎病病菌数量的变化,运用PCR-DGGE(Polymerase chain reaction-denatured gradient gel electrophoresis)技术分析棉花连作对土壤细菌群落结构和多样性的影响。结果表明,不同连作年限棉田的枯萎病菌和黄萎病菌量具有相似的变化特征,即在5~20年连作年限内,土壤中病原菌量随着连作年限的延长而增加,20年连作土壤病原菌数量达到最高峰;连作超过20年,土壤菌源数量开始下降。不同连作年限棉田土壤中枯、黄萎病病原菌的数量与发病株率呈显著正相关关系。连作棉田土壤细菌群落的DGGE图谱分析显示,连作棉田土壤细菌种群的组成、结构、丰富度均发生变化。连作5~40年的棉田土壤细菌种群Shannon-Wiener多样性指数(H')、均匀度(EH′)和丰富度(S)指数均降低,且连作年限越长降低越突出。     

Analysis of Cotton Secondary Metabolites under the Interaction between Verticillium dahliae and Upland Cotton

[Objective] The aim of this study is to investigate the cotton differential metabolites in the interaction between Verticillium dahliae and cotton plant, and to explore new clues for the further study on the defense mechanism of cotton against V. dahliae. [Method] To obtain the cotton root, stem and leaf samples of the pathogen treated and healthy controls, the upland cotton cultivar CCRI 24 was selected as tested cotton material in this paper, and inoculated with V. dahliae conidia or sterile water by injuring root at the 2-true-leaf stage. The 70% (volume fraction) methanol extracts of these samples were separated and detected by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS), and the metabolite data were obtained by the online XCMS software. Through multivariate statistics and student’s t-test, the differential metabolites were investigated. The types of metabolites were putatively identified based on the comparison of the experimental molecular mass and the monoisotopic accurate molecular mass of cotton metabolites. [Result] The efficiency of negative ion mode was higher than that of positive ion detection mode in the cotton metabolites in UPLC-ESI-MS analysis. 576 ions mainly distributed in cotton root were found in the cotton seedling tissues including root, stem and leaf, which were the distinguished differential metabolites between V. dahliae treatment and healthy control. Among them, 77 ions were identified as sesquiterpenoids, diterpenoids, flavonoids, carbohydrates, aliphatics and phenols. In addition to sesquiterpenes, 17 compounds, including caffeic acid, astragalin, isoastragalin, dillenetin, ent-catechin, gossypetin 8-rhamnoside, gossypicyanin, herbacetin 7-glucoside, leucocyanidin, quercetin 3'-glucoside, quercetin 3-glucoside, quercetin 7-glycosides, α,2',3,3',4,4',6-heptahydroxychalcone 2'-glucoside, melibiose, sucrose, sucrose 6-phosphate and 1-tetratriacontanol, had not been reported in the literatures on the interaction between V. dahliae and cotton, which may be the novel pathogenesis-related metabolites on cotton Verticillium wilt. [Conclusion] The putative pathogenesis-related metabolites of cotton Verticillium wilt may play an important role in the interaction of cotton defense against V. dahliae, which provides an important clue for exploring the new resistant mechanism of cotton to Verticillium wilt.     

Proteomics Analysis of Gossypium barbadense Leaves with Resistance to Verticillium dahliae under Infection Stress

[Objective] The purpose of this study is to analyze the resistance mechanism of island cotton under Verticillium wilt stress and to find possible resistance genes. [Method] The cotton resistance mechanism under the stress of Verticillium dahliae was studied by protein two-dimensional electrophoresis and mass spectrometry at the protein level. [Result] In the leaves, eleven proteins were down-regulated and 15 proteins were up-regulated after 2 hours of inoculation with Verticillium dahliae. The down-regulated proteins were mainly related to photosynthesis and carbon assimilation, and then we deduce that Verticillium dahliae is mainly broken cotton photosynthetic system to cause cotton susceptibility. The up-regulated proteins were mainly related to photosynthesis and benzoquinone reductase, beta-D-galactosidase, 14-3-3f protein and other disease-resistant protein. [Conclusion] It is speculated that the defense mechanism of island cotton on Verticillium wilt may occur at two levels: one is passive defense, it is manifested that the isoproteins of the down-regulated proteins such as chloroplast II AB binding protein and ribulose diphosphate carboxylase are highly expressed to maintain the stability of the island cotton photosynthetic system, the histone and 14-3-3f protein are highly expressed to keep the cell stability; the second is active defense, it is manifested that the high expressed β-D-galactosidase and benzoquinone reductase may be involved in the resistance of island cotton to Verticillium wilt.     

Effect of Pathogen Synergistic Pathogenicity on Severity of Verticillium Wilt of Cotton

Verticillium dahliae is a soil-borne and vascular pathogen causing Verticillium wilt of cotton, which is one of the most severe diseases affecting cotton production in China. Recently, a soil-inhabiting opportunistic pathogen was isolated and identified as Scopulariopsis gossypii; it could infect the host through wounds and increase disease severity in combination with V. dahliae. S. gossypii was able to form conidia in a nutritionally poor substrate and had similar biological characteristics to other vascular pathogens, while ultrastructure observation also confirmed that it was present in vascular tissue of cotton plants and produced conidia. Nematode activity in soil could cause wounds on the surface of cotton roots resulting in infection by the pathogens. On the basis of our analysis of the biological characteristics and infection process of S. gossypii and its interactions with other pathogens in cotton plants, we propose establishing a research program for multipathogen-host interactions in the cotton rhizosphere, which will enable detailed investigation of the synergistic pathogenesis of S. gossypii and V. dahliae.     

棉花黄萎病抗性与过氧化物酶同工酶分析

采用等电聚焦电泳方法 ,对棉花二对抗黄萎病近等基因系以及三对色素腺体近等基因系的子叶进行过氧化物同工酶 (POD)分析。结果表明 ,色素腺体和棉酚含量与棉花抗性没有必然的联系 ;环境条件对棉花抗性表现有一定的影响 ,但对子叶中 POD同工酶谱带无显著的影响 ;4条 POD同工酶酶带 (P4、P6、P7、P10 )的表现与棉花抗黄萎病性有关 ,这些酶带的特殊表现可用于棉花抗病性辅助鉴定和辅助育种     

棉花黄萎病及抗病育种研究进展

 介绍了黄萎病病菌致病机理、棉花黄萎病抗病机理及棉花抗黄萎病育种研究的现状,从不同的方面分析了制约棉花抗黄萎病育种研究的因素,提出了加快抗黄萎病育种研究进程的对策和建议。