重庆市豌豆枯萎病病原菌的分离与鉴定

张慧 马连杰 冯牧野

摘要：采用组织分离法，对重庆市豌豆枯萎病病原菌进行分离和纯化，结合形态学和基因序列分析对分离物进行鉴定，采用无伤接种法测定分离物对豌豆离体叶片、茎秆的致病性。结果表明，从枯萎病样品中分离获得5株菌株，菌落为圆形，初期为白色絮状，后期呈黄棕色至褐色;供试菌株的ITS序列和G3PDH基因序列与Ascochyta pinodes的同源性达到99%;致病性测定结果表明，供试菌株对豌豆叶片及茎秆均有致病性。表明引起重庆市豌豆枯萎病的病原菌为Ascochyta pinodes。

关键词：豌豆;豌豆枯萎病;病原鉴定;Ascochyta pinodes

中图分类号：S436.43         文献标识码：A

文章编号：0439-8114（2020）06-0084-04

DOI：10.14088/j.cnki.issn0439-8114.2020.06.016           开放科学（资源服务）标识码（OSID）：

Isolation and identification of pathogen of pea fusarium wilt in Chongqing city

ZHANG Hui，MA Lian-jie，FENG Mu-ye，LU Wen-cai，LIAO Dun-xiu

（Institute of Agricultural Resources and Environment，Chongqing Academy of Agricultural Sciences，Chongqing 401329，China）

Abstract： The tissue isolation method was used to isolate and purify the pathogen of spot of fusarium wilt of peas in Chongqing city， and the isolates were identified by combining with the analysis of morphology and gene sequence. The pathogenicity of the isolates to the leaves and stems of isolated peas was determined by the method of inoculation without injury. The results showed that five strains were isolated from infected leaves. The colony was round， white flocculent in the early stage and yellowish brown to brown in the later stage; The homology of ITS sequence and G3PDH gene sequences of the tested strain reached 99% with that of Ascochyta pinodes; The results of pathogenicity determination showed that the strain had pathogenicity to both leaves and stems of pea. It indicated that Ascochyta pinodes was the cause of pea blight in Chongqing city.

Key words： pea; pea fusarium wilt; pathogens identified; Ascochyta pinodes

豌豆是中国食用豆的一个主要品种，中国豌豆生产量占全球豌豆总产量的60%以上。中国是第二大干豌豆生产国家，2013年干豌豆产量达1 566亿t。在生产上，豌豆病害是制约产量提高的重要因素。已报道的豌豆病害有25余种[1]，而豌豆壳二孢枯萎病（常被称为黑斑病）是豌豆最严重的病害之一，分布于世界各地，包括几乎所有种植豌豆的区域[2]。该病菌可为害植株的根、茎、叶和豆荚，造成豌豆产量减少10%～60%[1，3]。壳二孢枯萎病是由Ascochyta pinodes（有性态Mycosphaerella pinodes）、Phoma medicaginis、Ascochyta pisi（有性态Didymella pisi）和Phoma koolunga复合侵染引起的真菌病害[4，5]。这种枯萎病复合物引起一系列不同的症状，包括黑斑、脚腐、茎腐、叶片和荚果斑点。Ascochyta pinodes可以侵染豌豆植株的幼苗和所有地上部分，引起壞死性叶斑，茎基部变黑，幼苗蛀烂。由Phoma pinodella引起的症状与Ascochyta pinodes症状相似。然而，Phoma pinodella侵染可导致更严重的基部腐烂症状，可以延伸到地面以下，同时对叶、茎和荚果造成的损害较小。……