梨胶胞炭疽病菌的分离、鉴定及其生物学特性

近年来,由胶胞炭疽病菌(Colletotrichum gloeosporioides)引起的梨炭疽病在江苏、安徽两省酥梨产区普遍发生。本研究采集病样,分离并鉴定梨胶胞炭疽病菌25株。采用梨果表面刺伤后接种菌块的方法,观察炭疽病菌对砀山酥梨的致病性。平板接种炭疽病菌菌块,测试不同培养温度、pH值、碳源、氮源对炭疽病菌菌丝生长的影响。结果表明:参试的25株炭疽病菌中3株致病性较强,18株致病性中等,4株致病性较弱。致病性强的菌株其菌落颜色较深,菌丝浓密;致病性弱的菌株其菌落颜色均为白色,菌丝稀疏。菌落生长快,菌株的致病性较强;菌落生长慢,菌株的致病性较弱。菌株的产孢能力和致病性之间无相关性。梨胶胞炭疽病菌最适生长温度为25～30℃,最适生长pH值为5.0～7.0;菌丝对多种单糖和双糖等碳源及有机氮、无机氮均可利用,最适碳源为蔗糖,最适氮源为牛肉浸膏。

橡胶炭疽病菌致病相关突变体的筛选及表型分析

通过对实验室已获得的1 685个橡胶炭疽病菌T-DNA插入突变体的生物学及致病性测定,从中筛选出15个致病力明显减弱的突变体,并进行PCR检测。结果表明,该基因组中均含潮霉素磷酸转移酶基因片段。在PDA培养基上,其中2个突变体菌落颜色异常,6个生长速率下降,8个产孢量显著降低,2个突变体分生孢子形态异常,2个附着胞形态异常,3个不能形成附着胞,在洋葱表皮上,侵染钉形成率显著降低。

橡胶树胶孢炭疽病菌Cg4LysM基因对菌丝生长的影响

本研究通过PT-PCR技术克隆了橡胶树胶孢炭疽病菌的1个候选效应蛋白基因并命名为Cg4LysM。该基因含1 983 bp的完整开放阅读框(ORF),编码660个氨基酸的多肽。生物信息学分析结果表明,该蛋白含有4个LysM保守结构域,不含任何跨膜结构域,其1~20位氨基酸是典型的信号肽序列,推测为胞外分泌蛋白。氨基酸序列比对结果表明,Cg4LysM与草莓胶孢炭疽菌(Colletotrichum gloeosporioides Nara gc5)、高粱炭疽病菌(Colletotrichum sublineola)、石膏样小孢子菌(Microsporum gypseum CBS)的LysM蛋白同源,同源性分别为97%、59%、55%。此外,本研究根据同源重组的原理构建了该基因的敲除突变体△Cg4LysM,进一步分析发现,△Cg4LysM的生长速度慢于野生型菌株,暗示Cg4LysM与胶孢炭疽菌的生长有关。

枯草芽胞杆菌sf 628对梨炭疽病的控制作用

近年来,由胶胞炭疽病菌Colletotrichum gloeosporioides引起的梨炭疽病在砀山酥梨产区普遍发生。为探讨生防菌对梨炭疽病的控制作用,采用平板对峙生长法和喷雾法研究了枯草芽胞杆菌sf 628对梨炭疽病的室内外防治效果。枯草芽胞杆菌sf 628发酵菌液、100倍和500倍稀释液对胶胞炭疽病菌菌丝生长的平板抑制率分别为91.45%、88.80%和86.63%。梨果先喷施生防菌液sf628,24 h后接种炭疽病菌,对梨炭疽病的防治效果为83.74%;梨果先接种炭疽病菌,24 h后再喷施生防菌液sf 628,对梨炭疽病的防治效果为56.82%。生防菌sf 628可湿性粉剂100、1000和2000倍稀释液对梨炭疽病的田间防治效果分别为75.89%、60.78%和50.60%,常规对照药剂多菌灵的防治效果为61.10%。研究表明,枯草芽胞杆菌sf 628是一株对梨炭疽病有较好控制作用的生防菌。

Isolation and Identification of Colletotrichum gloeosporioides in Pears and Its Biological Characteristics

[Objective] This study aimed to explore the biological characteristics of Col etotrichum gloeosporioides in pears. [Method] Twenty-five C. gloeosporioides strains were isolated and identified from the diseased samples. Their pathogenicity was identified by inoculating the surface of punctured pears with fungal discs. The effects of different temperatures, pH values, carbon sources and nitrogen sources on the growth of C. gloeosporioides mycelia were explored by incubating fungal discs on the center of plates. [Result] Among the twenty-five C. gloeosporioides strains, three had strong pathogenicity, and eighteen had intermediate pathogenicity, and four strains had weak pathogenicity. Those highly-pathogenic strains had darker colonies, with dense mycelia, whereas those lowly-pathogenic ones had white colonies, with sparse mycelia. Those with fast-growing colonies showed strong pathogenicity, while those with slowly-growing colonies displayed weak pathogenicity. There was no relationship between conidia yield and pathogenicity. The optimum temperature for the growth of C. gloeosporioides mycelia was 25-30 ℃, and the optimum pH was 5.0-7.0. C. gloeosporioides could make use of various carbon sources （monosaccharide and disaccharide）, inorganic and organic nitrogen sources, and the optimal carbon source and nitrogen source were sucrose and beef extract, respectively. [Conclusion] Our study benefits further understanding of C. gloeospori-oides and helps to control pear anthracnose more effectively.