猕猴桃灰霉病病原菌鉴定及室内药剂筛选

通过病原菌形态特征、致病性测定及rDNA-ITS的同源性分析,确定江西奉新猕猴桃灰霉病的病原菌为Botrytis cinerea(有性态Botryotinia fuckeliana),该病菌在5~30 ℃的温度范围内均能正常生长,且该病菌只能通过伤口接种引起猕猴桃发病。采用菌丝生长速率法测定11种杀菌剂对该病原菌的毒力大小,结果表明,肟菌·戊唑醇水分散粒剂、氟硅唑乳油、异菌脲悬浮剂、苯醚甲环唑水分散粒剂、戊唑醇悬浮剂和吡唑醚菌酯乳油对其菌丝生长有较强的抑制作用,EC50分别为0.118 0、0.288 6、0.355 2、1.541 8、1.760 2和1.778 4 μg/mL。     

Genetic diversity of Botrytis in New Zealand vineyards and the significance of its seasonal and regional variation

Species‐ and population‐specific differences in fungicide resistance and aggressiveness within Botrytis makes basic data on genetic diversity important for understanding disease caused by this fungus. Genetic diversity of Botrytis was surveyed between 2008 and 2012 from grapes from five New Zealand wine‐growing regions. A total of 1226 isolates were gathered from symptomless flower buds at the start of the growing season and 1331 isolates from diseased fruit at harvest. Two species were found, B. cinerea and B. pseudocinerea. Botrytis pseudocinerea was common in both Auckland vineyards sampled, and infrequent elsewhere. However, even in Auckland, it was rarely isolated from diseased fruit. The presence of the Boty and Flipper transposons was assessed. Isolates with all four transposon states (Boty only, Flipper only, both Boty and Flipper, no transposons) were found for both species. Both vineyards in the Auckland region had high numbers of Flipper‐only isolates at flowering; both vineyards from the Waipara region had high numbers of Boty‐only isolates at flowering. Most isolates from diseased fruit at harvest contained both transposons. These observations suggest that B. pseudocinerea, and isolates with one or both of the transposons missing, may be less aggressive than B. cinerea, or than isolates with both transposons present. Two clades were resolved within B. pseudocinerea, only one of which has been reported from European vineyards. Phylogenetic diversity within B. cinerea in New Zealand was similar to that known from Europe, including isolates that appear to match Botrytis ‘Group S’. The taxonomic implications of this genetic diversity are discussed.     

枯草芽孢杆菌B21抗菌蛋白的分离纯化及特性研究

枯草芽孢杆菌(Bacillus subtilis)B21对番茄灰霉病菌(Botrytis cinerea Pers.)有较强的抑制作用,其产生的抗菌物质主要是抗菌蛋白.为明确该抗菌蛋白的理化性质,利用饱和硫酸铵沉淀法得到拮抗粗提蛋白,所获粗提蛋白不耐高温,对胰蛋白酶和蛋白酶K部分敏感,对链霉蛋白酶不敏感而对氯仿敏感,作...     

灰葡萄孢霉高效拮抗木霉菌株的筛选及其翻译延伸因子序列分析

为进一步开发利用木霉菌资源,对从菜田土壤中初步分离纯化获得的12株木霉菌株,采用对峙培养和生长速度测定法进行灰葡萄孢霉(Botrytis cinerea)高效拮抗木霉菌株的筛选,并通过翻译延伸因子序列同源性比较对其进行分子鉴定。结果表明菌株Tr-9701和Tr-1108的生长速度快,对病原菌的抑制率高,且协同应用有一定的增效作用。经对Tr9701和Tr1108翻译延伸因子同源序列分析,并结合其形态特征结果表明,Tr9701为绿色木霉(Trichoderma viride)和Tr1108为深绿木霉(Trichoderma atroviride)。绿色木霉和深绿木霉为菜田生境习居菌,2种菌株协同利用对蔬菜灰霉病有较好的协同增效作用,应进一步设计利用多靶位木霉菌来提高防病效果。     

Evaluation of anilinopyrimidine and other fungicides for control of grey mould (Botrytis cinerea) in container-grown Calluna vulgaris

BACKGROUND:Grey mould (Botrytis cinerea Pers. ex Fr.) is a damaging disease affecting container-grown Calluna vulgaris (L.) Hull. Trials were conducted to test anilinopyrimidine fungicides (pyrimethanil, cyprodinil and mepanipyrim), azoxystrobin and tolylfluanid for crop safety on cultivars, and to evaluate spray programmes based on alternating these fungicides with different modes of action for disease control. Timing and application at different spray intervals were also investigated. RESULTS:Cyprodinil was slightly phytotoxic on only one of the seven cvs tested. Alternating nine-spray programmes of azoxystrobin with an anilinopyrimidine or tolylfluanid, or an anilinopyrimidine with tolylfluanid, with the first spray applied immediately after potting, were equally effective in reducing disease incidence and severity compared with untreated plants. Spray programmes of five or eight sprays, using an anilinopyrimidine and at least one other fungicide with a different mode of action, applied alternately or in blocks of two sprays of the same fungicide and immediately after potting, were equally effective in reducing disease. Omission of early fungicide sprays until 9 weeks after potting and subsequent alternate application of pyrimethanil and tolylfluanid with a different mode of action at 6 week intervals gave reduced disease control. CONCLUSION:There is potential to use spray programmes based on alternating anilinopyrimidines with fungicides with a different mode of action to provide effective control of B. cinerea on C. vulgaris.     

来源于植物的短肽LD-1对番茄灰霉病的抑制作用及其抗病性研究

短肽LD-1是从牛心朴子草根中新分离的具有抗菌活性的多肽。本文以番茄灰霉病为供试菌株,研究了短肽LD-1对番茄灰霉病的抑制作用及其抗病性。结果表明:短肽LD-1对番茄灰霉病菌菌丝生长及孢子萌发具有较好的抑制作用。离体试验中,短肽LD-1对番茄灰霉病菌菌丝生长抑制作用的EC50在24、48、72h分别为28.3051、30.2134、32.7427mg/L,对孢子萌发的EC50为16.6919mg/L;活体试验中,短肽LD-1对番茄灰霉病具有较好的防治效果,90mg/L短肽LD-1对番茄灰霉病防治效果达到74.24%。短肽LD-1能导致病菌芽管发育畸形,不能正常发育。短肽LD-1对番茄种子的萌发和根系生长具有明显的促进效果,并且经短肽LD-1处理后的番茄须根发达,须根数目明显增多,促进了番茄的生长发育,提高了抗病性。     

Screening of conidium development mutant of Botrytis cinerea and functional analysis of the related gene

A novel conidium development mutant was obtained by screening the transformants of Botrytis cinerea produced by Agrobacterium tumefaciens mediated method, which lost the ability of producing conidia. The flanking sequence of T-DNA insertion site was acquired by TAIL-PCR technology, and then, the T-DNA insertion in the second exon of BC1G_02800.1 confirmed by BLAST between the flanking sequence and the known sequence in the B. cinerea gene database. The mutant gene was identified as BC1G_02799.1 located in the upstream of BC1G_02800.1 gene by RTPCR. The DNA full-length sequence of BC1G_02799.1 was 1951 bp and contained 1848 bp coding region, which encoded a 615 amino acids putative protein similar to ABC-transporter, and the function of BC1G_02799.1 gene was unknown to date. Phenotype analysis of the mutant found that the mutant strain colony was white, grew slowly, and did not produce conidium and sclerotia on PDA medium but showed a stronger pathogenicity to tomato leaves and successfully increased the enzyme activity related to pathogenicity compared to the wild type strain. The results suggested that the BC1G_02799.1 gene was involved in the conidium development, the sclerotia formation, and pathogenicity in B. cinerea. Our research will facilitate in understanding the molecular mechanism of conidium development, sclerotia formation, and pathogenic in B. cinerea.     

非洲菊灰霉病抗性鉴定技术的研究

 灰霉病是非洲菊切花破坏性的病害,在采后阶段的发生和危害尤其严重,培育抗性品种具有重要意义,抗性非洲菊材料的筛选鉴定是抗性品种培育的必要技术。以生产中显著抗病和感病品种为试材,研究非洲菊灰霉病抗性鉴定技术中适宜的试验材料、灰霉菌孢子浓度、孢子侵染条件和病害评估方法。结果表明, 非洲菊花瓣正面接种病害发展快、不同抗性品种间差异明显;分生孢子的侵染萌发需要充足的氧气和营养,2μL液滴、马铃薯葡萄糖的添加使花瓣发病整齐均匀;病菌的增殖危害与接种时分生孢子和马铃薯葡萄糖的初始浓度相关,在一定浓度范围内,二者呈反比关系,其中分生孢子适宜浓度为3.0×105~1.0×106个孢子/mL,马铃薯葡萄糖的适宜浓度为3%~6%;接种花瓣放置于室温、自然光、近100%湿度条件下,接种48h是最佳的评估时间。     

Multiple-disease resistance in Vicia faba: Multi-environment field testing for identification of combined resistance to rust and chocolate spot

The aim of this work was to identify Vicia faba germplasm resistant both to rust and chocolate spot. A collection of 43 accessions of V. faba previously identified as chocolate-spot resistant was evaluated for rust and chocolate spot resistance in Egypt and in Spain. The genotype and genotype × environment (GGE) biplot analyses allowed the selection of 11 accessions resistant to both diseases in those locations. These 11 accessions were evaluated for rust in an additional field trial in Spain, all performing better than the susceptible check. The joint analysis of the 11 accessions in the four field trials where they had been evaluated revealed no significant effects either for genotype, environment or the genotype × environment interaction. They are hence promising sources of resistance, both for their low severities and their stable responses across the studied environments. This collection was also tested under controlled conditions against the Egyptian and the Spanish isolates of rust present in the field trials further underlining the stable performance of these genotypes. Finally, the effect of previous infection with chocolate spot on rust resistance was assessed under controlled conditions and no influence of it was found.     

魔芋灰霉病致病菌及其生理特性的研究

为探明魔芋灰霉病的发病规律,对其发病症状、致病菌及其生理特性进行了较系统的研究。结果表明,魔芋灰霉病菌为葡萄孢菌属(Botryotinia);其分生孢子萌发的温度范围为10~30℃,最适温度为20~25℃;在pH 4~9条件下均能萌发,最适pH为5;在各种营养液中均能萌发,在2%的蔗糖液中萌发最好,其次为2%的葡萄糖和魔芋甘露低聚糖;分生孢子的致死温度为57℃、5 min。魔芋灰霉病菌菌丝在4种培养基上均能生长,其中PDA培养基最适宜菌丝生长,最适的产孢培养基为PDA+魔芋甘露低聚糖;菌丝在5~30℃均能生长和产孢,适宜温度为20~25℃;在pH 4~9均能生长及产孢,最适pH为5。