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[Objective] The aim of this study is to make use of the antagonism and growth promoting ability of Bacillus methylophicus strain LH-L3, and add it into the chemical seed-coating agent as an active ingredient. [Method] The optimum formula (agent No.4: 2.5% fludioxonil＋10% thiram＋10% carbofuran＋109 cfu·mL－1 LH-L3) was obtained based on the sterilization effect of four seed coating agents on infected seed. Then the survival ability of the strain in agent No.4 and its shelf life were identified by plate test. Finally, the disease and pest control effects were tested in the greenhouse pot experiment. [Result] The results showed that the agent No. 4 had the best control effect, and 1.1×105 cfu·mL－1 of LH-L3 after storage at room temperature for 6 months. Compared with the control, the formula No. 4 increased 11.54% emergence rate, 18.74% plant type height and 20.91% leaf width, respectively, in the 15th day after emergence, and decreased 65.42% and 48.56% of Verticillium wilt at day 2 and day 5 after the disease occurred. The cotton seeds coated with formula No. 4 reduced 83.07% of plant and 92.33% of leaf damaged by Sylepta derogata, compared with the negative control without seed coating. [Conclusion] Strain LH-L3 and chemical pesticide compound as a seed coat agent is completely feasible. And the developed biological complex seed-coating agent has better control effect on Verticillium wilt and Sylepta derogate. 相似文献
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Induction of Defense Enzymes and Control Effect of Endogenous and Rhizosphere Bacteria on Verticillium Wilt of Cotton 总被引：1，自引：1，他引：0
《棉花学报》2019,31(6):550-558
[Objective] The colonization in soil, promoting cotton growth effects, inducing resistance of cotton and controlling effects to cotton Verticillium wilt of Bacillus vanillea SMT-24, B. velezensis BHZ-29, B. subtilis SHT-15 and B. atrophaeus SHZ-24 were studied in this paper, which provides a scientific basis for better control of cotton Verticillium wilt. [Method] The antagonistic resistance of these bacteria to Verticillium dahliae was judged by observing the inhibition zone; the number of colonization in the rhizosphere soil was tested to represent the colonization ability of antagonistic bacteria; the growth promoting effect of strains on cotton was determined by analyzing cotton plant height, root length, root hair number and number of leaves; the effect of antagonistic strains on cotton induced resistance was explored by determining catalase (CAT), superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidase (POD) activities in cotton leaves, and investigating the disease index of cotton after inoculation with antagonistic bacteria solutions. [Result] SMT-24, BHZ-29, SHT-15 and SHZ-24 strains significantly inhibited the growth of V. dahliae and formed a clear zone of inhibition. After 2 to 22 days of the third inoculation, these strains of bacteria could survive in the root soil, increase the plant height, leaf number, and root hair number of cotton plants, as well as the activities of cotton CAT, SOD, PAL and POD enzymes, but reduced the disease index. [Conclusion] SMT-24, BHZ-29, SHT-15 and SHZ-24 strains can promote cotton growth, increase the activities of some related defense enzymes, and effectively control cotton Verticillium wilt. 相似文献
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棉花黄萎病菌与品种互作的格局分析 总被引：2，自引：1，他引：1
以23个棉花品种(系)接种18个黄萎病菌株的抗感表现(病情指数)为分析对象,在聚类分析的基础上用双标图方法研究了寄主 病原的互作格局。结果表明：23个棉花品种分为6个抗病性不同的品种组,18个菌株聚为5个致病力不同的菌株群。5031和50312组成的品种组G1对所有菌株群具有最高抗性,可作为抗黄育种的高抗质源。来自河北成安的VD25与来自江苏常熟的XS4、XS6和XS7组成的菌株群S4与G5品种组存在互作,其致病力最强,是棉花黄萎病的优势菌株群,在抗黄育种和抗黄鉴定中应予以高度重视。S2和S3菌株群与56%的参试品种构成的3个品种组间存在互作,是潜在的优势菌株群。具平均抗病性的G2品种组是黄萎病菌的优良寄主。 相似文献
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在棉花现蕾期筛选到一个高抗黄萎病的内生菌菌株,根据16S rDNA序列同源性,将其命名为Paenibacillus xylanilyticus YUPP-1(解木聚糖类芽孢杆菌YUPP-1)。当把该菌标记后,每株棉花施用该菌株达到1×106 cfu时,该菌能顺利定殖于棉花体内。室内盆栽抗病实验表明：当生防菌灌根处理的菌量为每株1×106,1×107,1×108 cfu,其在结铃期的发病株率仅为8%,5%和2%,而此时对照的发病株率高达96%。2008－2009年的小区防治结果表明：当生防菌灌根处理的菌量为每株1×108 cfu时,2008年处理区的棉花黄萎病发病株率和病指为14.8%和8.5,对照分别为53.3%和26.5,2009年处理区的棉花黄萎病发病株率和病指为9.4%和6.5,对照为37.8%和18.8。这些结果表明,解木聚糖类芽孢杆菌YUPP-1对防治棉花黄萎病具有巨大的应用潜力。 相似文献
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[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. 相似文献
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Li Shezeng Niu Luxin Li Bochao Chen Xiuye Liu Chang Lu Xiuyun Guo Qinggang Ma Ping Ma Zhiying 《棉花学报》2021,32(6):501-521
[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. 相似文献
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棉花黄萎病菌拮抗细菌TUBP1的分离鉴定及其防病作用 总被引：1，自引：1，他引：0
为研制和开发新疆棉花黄萎病优良生防菌剂,采用琼脂平板扩散法,通过初筛和复筛,分离得到一株对棉花黄萎病菌(Verticillium dahliae)具有较强抑菌活性的拮抗菌株TUBP1。通过对形态观察、生理生化实验、16S rDNA和gyrB的基因序列分析,确定该菌株为Bacillus axarquiensis;利用生长速率法和牛津杯法分别测定了发酵液对棉花黄萎病菌抑菌活性,对棉花黄萎病菌菌丝生长抑制率为78.62%±3.48%,抑菌圈直径为21.00±3.86 mm。温室盆栽试验中,TUBP1最大生防效果为43.67%,小区试验中,TUBP1最大生防效果为43.07%。 相似文献
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生防细菌NCD-2突变体构建及抑菌功能基因的防病作用 总被引：2，自引：1，他引：2
生防细菌NCD-2是一株枯草芽孢杆菌菌株,该菌株通过分泌抑菌肽而对棉花黄萎病病原菌和棉花立枯病病原菌起到抑制作用。本研究着重通过原生质体法与诱导转座方法,建立了携带转座子Tn917质粒pTV1对枯草芽孢杆菌NCD-2野生菌株的转化体系与转座子突变技术,获得1500多个转座子插入突变子。通过测定这些突变子对大丽轮枝菌的抑制作用,筛选到2个抗生作用丧失的抑菌功能缺失的突变子。室内盆栽试验结果表明这2个抑菌功能缺失突变子对棉花立枯病的防效显著低于野生菌株,说明NCD-2野生菌株产生的抑菌肽在该菌株防治棉花立枯病中起到主要作用,进而说明编码该抑菌肽的基因在该菌株防治棉花立枯病中具有重要作用。 相似文献
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棉花黄萎病菌与抗黄萎病遗传育种研究进展 总被引：21，自引：1，他引：21
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 相似文献
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[Objective] The study of pathogenicity differentiation of Verticillium dahliae in the cotton growing areas in Hunan province could provide theories for breeding and promoting resistant varieties as well as integrated control of Verticillium wilt of cotton. [Method] Seventy seven strains of V. dahliae from major cotton growing areas in Hunan province were evaluated their cultural characteristics. The growth rate, sporulation quantity and pathogenicity of 31 strains among them were tested and SAS 9.1.3 statistical analysis software was used to cluster the pathogenicity of the tested strains. The special primer (D-1/D-2 and ND-1/ND-2) for polymerase chain reaction assay was used to test pathotype of 24 strains. [Result] According to their colony morphology on potato dextrose agar media and culture characteristics, the strains were classified into the sclerotium type, the hyphal type and the intermediate type accounted for 14.28%, 42.86% and 42.86%, respectively. The growth rate and sporulation quantity of the strains were 1.22–2.54 mm·d－1 and 5.3×106–40.6×106 spore·mL－1, which showed significant difference. In addition, based on the clustering results of average disease index,the strains were clustered into three groups which showed high (Type Ⅰ), low (Type Ⅱ) and moderate (Type Ⅲ) virulence to cotton with the proportion of 3.2%, 12.9% and 83.9%, respectively. [Conclusion] It is conclude that the main culture types of V. dahliae in Hunan province are the hyphal type and the intermediate type, and the strains showed pathogenicity differentiation. The result showed that defoliating pathotype of V. dahliae was dominant in the cotton growing areas of Hunan province． 相似文献
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[Objective] Our research aimed to identify pathogenicity defective mutants from a T-DNA insertion mutant library of Verticillium dahliae strain Vd991 and to analyze pathogenicity-related genes. [Method] In total, 294 T-DNA insertion mutants of V. dahliae were tested for their virulence using a cotton infection assay. Southern blot assays were performed to identify the T-DNA insertion copy number of each pathogenicity defective mutant. DNA sequences flanking the T-DNA insertional sites of each mutant were analyzed by high-efficiency thermal asymmetric interlaced PCR. [Result] Based on the virulence assay, the disease indices of cotton plants inoculated with each mutant decreased very significantly in comparison with the index of those inoculated with Vd991. The Southern blot assay revealed that only one mutant contained two T-DNA insertions, while the remaining eight mutants harbored a single T-DNA insert. An analysis of biological characteristics found that the growth and conidial production of these mutants were impaired by the T-DNA insertions compared with the wild type Vd991. The T-DNAs' insertion position and distribution in each mutant were identified by comparison with genome sequences of strain VdLs.17. Furthermore, the pathogenicity-related genes were cloned from strain Vd991. [Conclusion] The screening and identification of T-DNA insertion mutants is an effective method to identify the pathogenicity-related genes of V. dahliae on a genome-wide scale. This laid a foundation for the further breeding of disease-resistant cotton varieties and will promote the study of the pathogenic molecular mechanisms of V. dahliae. 相似文献
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Physiological Mechanism of Different Varieties and Potassium Application Amounts on Cotton Resistance to Verticillium Wilt 总被引：1，自引：0，他引：1
[Objective] It has theoretical guiding significance for the comprehensive control technology of Verticillium wilt of cotton to explore the effect of potassium nutrition on mechanism of cotton Verticillium wilt resistance under the condition of pathogen stress of Verticillium dahliae. [Method] Three cotton cultivars Jimian 11 (JM-11 as susceptible cultivar), Zhongzhimian 2 (ZZM-2 as control cultivar), Nongda 601 (ND-601 as resistant cultivar) were adopted, and each cultivar had 0, 75, 150, 225, 300 kg· hm－2 potassium levels represented by K0, K75, K150, K225, K300. Two factors split plot experiment was designed under artificial nursery inoculated Verticillium dahlia, to observe disease index of Verticillium wilt, and researched resistance associated substances content and enzyme activity. [Result] (1) The lignin, total phenol contents and POD, PAL activity in leaves or roots of JM-11 were more greatly influenced by Verticillium wilt compared with ZZM-2, ND-601; the crucial disease resistance substances and enzymes were lignin, total phenol and PAL for JM-11, total phenol for ZZM-2 and ND-601. (2) The lignin, total phenol contents and POD, PAL activities of JM-11 were more sensitive to root potassium than ND-601, but weaker than that of ZZM-2. The sensitivity of POD, PAL activities in functional leaves of JM-11 to potassium were more sensitive than ND-601 and ZZM-2. (3) The best effect of potash fertilizer for relieving disease index for JM-11, ZZM-2, ND-601 were 32.5%, 26.4%, 16.5%, and corresponding potassium application amounts were 290.5 kg·hm－2, 253.5 kg·hm－2 and 229.5 kg·hm－2, respectively. [Conclusion] Application of potassium could increase the resistance to Verticillium wilt and reduce the occurrence of cotton Verticillium wilt. 相似文献
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为了研究新疆乌苏地区棉花黄萎病菌的致病型和群体间的变异性,从该地区不同棉株上分离8株病原菌,通过对病原菌的菌落培养形态、菌丝和孢子的显微结构、对寄主的致病性、ITS序列的克隆、菌株间的系统进化及营养亲和性分析等方面进行了研究。结果表明：分离的病原菌均属于非落叶型棉花黄萎病菌(Verticillium dahliae),为大丽轮枝菌;系统进化树显示8株菌在系统进化上属于2个不同的进化方式;8株黄萎病菌在相同的寄主上致病性存在差,Vd-1菌株致病力最强,Vd-34致病力最弱;不同致病力的菌株之间的营养亲和性分为2个营养亲和群(VCGs)。新疆乌苏地区棉花黄萎病的致病菌是大丽轮枝菌,病原菌在进化上差异显著。 相似文献
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大丽轮枝菌拮抗细菌菌株12-51的筛选鉴定与抗菌物性质分析 总被引：2，自引：0，他引：2
由大丽轮枝菌（Verticillium dahliae Kleb）引起的棉花黄萎病,是一种危害严重的世界性病害。通过初筛和复筛得到了对棉花黄萎病具有较高拮抗活性的细菌菌株12-51,通过对其进行形态鉴定、生理生化特征鉴定和16S rDNA全序列分析,最终鉴定此菌株为Bacillus velezensis,16S rDNA序列相似度达99.74%。采用有机溶剂萃取和盐析法对发酵产物进行了提取,初步断定拮抗物质为蛋白类。通过对粗蛋白进行进一步研究发现,60%组分活性较高,并且对热以及酸碱敏感性较小。试验结果为棉花黄萎病的生物防治奠定了基础。 相似文献
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《棉花学报》2018,30(2):136-144
[Objective] The goal of this experiment was to study the effects of Verticillium wilt on photosynthetic characteristics and chlorophyll fluorescence characteristics of cotton seedlings, and to provide a theoretical basis for the identification of cotton resistance to Verticillium wilt at the seedling stage and the establishment of Verticillium wilt control techniques. [Method] Zhongzhimian 2 (Verticillium wilt resistant cultivar, ZZM-2) and Jimian 11 (Verticillium wilt susceptible cultivar, JM-11) were selected. Verticillium dahliae was inoculated at the two true leaves stage. The photosynthetic and chlorophyll fluorescence parameters of the last fully expanded young leaves were measured at 6 d after inoculation. [Result] (1) Under the stress of Verticillium wilt, the leaf area per plant, leaf water potential, photosynthetic pigment content and net photosynthetic rate (Pn) of cotton seedlings decreased, and the decreases in ZZM-2 were smaller than those in JM-11. (2) The decreased intercellular CO2 concentration and increased stomatal limitation value in the two test materials indicated that the decrease in Pn was mainly caused by stomatal limitation, and the decrease in chlorophyll a in ZZM-2 was smaller than the decrease in chlorophyll b, whereas the opposite trend was observed in JM-11. (3) The light response curve parameters of ZZM-2 were increased and theCO2 response curve parameters of ZZM-2 were decreased, but all parameters in JM-11 were decreased. For ZZM-2, the potential photochemical efficiency (Fv/Fm), the quantum yield of regulated energy dissipation (YNPQ) and the quantum yield of non-regulated energy dissipation (YNO) were increased slightly and the actual photochemical efficiency (YII) was decreased slightly, but the differences in these four indexes between diseased plants and healthy plants was not significant; however, the variable fluorescence yield (Fv) was significantly decreased. For JM-11, Fv, Fv/Fm, YII and YNPQ were significantly decreased, and YNO was significantly increased. (4) Fv/Fm in leaf lesion sites decreased, but that in non-lesion sites did not decline or even showed a slight increase. [Conclusion] Verticillium wilt resistant cultivars with higher photosynthetic capacity and carbon assimilation ability could reduce the effects of Verticillium wilt on photosynthetic characteristics and reduce the impacts on cotton yield and quality. 相似文献
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为了筛选可指示Verticillium dahliae致萎毒素毒力大小的指示菌,并对致萎毒素抑制指示菌生长的机理进行初步探讨,采用指示菌平板打孔法,对黑曲霉(Aspergillus niger)BF-3758、枯草芽孢杆菌(Bacillus subtilis)、金黄色葡萄球菌(Staphylococcus aureus)、啤酒酵母(Saccharomyces cerevisiae)等12种具有微生物种属代表性的菌株进行筛选;采用拮抗试验、致萎试验结合显微观察的方法,初步探索了Verticillium dahliae致萎毒素对指示菌生长的抑制作用。结果表明,Verticillium dahliae粗毒素对Aspergillus niger BF-3758的生长具有较强的抑制作用,且该抑制作用和粗毒素对离体叶片的致萎作用具有一致性,并存在一定的量效关系,Aspergillusniger BF-3758菌株可以作为Verticillium dahliae致萎毒素的毒力指示菌;Verticillium dahliae粗毒素可抑制Aspergillus niger BF-3758的孢子萌发和菌丝生长,对成熟菌丝也有破坏作用。上述研究可为评价Verticillium dahliae的致病力提供一种操作简便、耗时短、费用低的生物学方法。 相似文献