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林业科学 ›› 2000, Vol. 36 ›› Issue (2): 75-81.doi: 10.11707/j.1001-7488.20000212

• 论文及研究报告 • 上一篇    下一篇

树木溃疡病病原真菌类群分子遗传多样性研究Ⅱ——Botryosphaeria 属28S rDNA-PCR-RFLP和RAPD解析

张星耀 赵仕光1 吕全 贾秀贞 刘会香2   

  1. 中国林业科学研究院森林生态环境与保护研究所,北京100091;山东农业大学林学院,泰安271000;1.西北林学院讲师,在职博士研究生。;2.山东农业大学助教,客座研究人员。
  • 收稿日期:1999-10-14 修回日期:1900-01-01 出版日期:2000-03-25 发布日期:2000-03-25

MOLECULAR GENETIC DIVERSITY OF PATHOGENIC FUNGAL GROUP CAUSING TREE CANKER Ⅱ——28S rDNA-PCR-RFLP AND RAPD ANALYSIS OF BOTRYOSPHAERIA SPP

Zhang Xingyao,Zhao Shiguang,Lü Quan,Jia Xiuzhen,Liu Huixiang   

  1. The Research Institute of Forest Ecology, Environment and Protection, CAF Beijing 100091;Forestry College of Shangdong Agriculture University Taian 271000
  • Received:1999-10-14 Revised:1900-01-01 Online:2000-03-25 Published:2000-03-25

摘要:

Botryosphaeria Ces. Et de Not .(无性阶段是Dothiorella Sacc .)属病原真菌能引起多种林木溃疡病的发生,在我国分布区域十分广泛。病原菌因较大的寄主范围和地理分布区域表现出明显的分子遗传分化,通过对来自5个区14个寄主种(品种)的30个菌株进行28SrDNA-PCR-RFLP和RAPD解析,结果表明,所有供试菌株都能扩增出稳定一致的28SrDNA条带,6种内切酶不能区分该属内部的28SrDNA差异。RAPD结果表现出该属丰富的多样性(92.81%) ,在0.845相似系数的水平上所有菌株被识别为13类,其中有地理分化、寄主分化的表现,也有不表现地理、寄主分化的。从DNA水平上证实引起雪松溃疡病的病原为B.dothidea,表明B.dothidea不仅能够寄生被子植物,还能寄生裸子植物,有非常宽阔的寄主范围。随机扩增引物K16和R14可以作为陕西菌株和湖南菌株与其它地区菌株区别的鉴别性引物。传统上认为苹果轮纹病菌(B.berengeriana de Not.f.sp .piricola (Nose)Koganezawa et Sakuma)为苹果干腐病菌(B.berengeriana be Not.)的专化型,它们与杨树溃疡病原分属两个种。本试验分析表明这种分类较为合理。

关键词: Botryosphaeria 属, 28SrDNA-PCR-RFLP, RAPD, 鉴别性引物

Abstract:

Botryosphaeria Ces. et de Not., its a sexual phase is Dothiorella Sacc., causes canker of many varieties of trees and its distribution region is very wide. These two agents cause molecular gene diversity. PCR-amplified 28S rDNA products of 30 isolates from 5 regions and 14 varieties of host trees were the same. The polymorphism on digestion with six restriction enzymes was nearly the same. So, the 28S rDNA-PCR-RFLP analysis was not suitable for differentiating the species within genera. RAPD analysis of all the isolates showed that there was abundant polymorphic DNA within the genera(92.81%). All the tested isolates were divided into 13 clusters according to 0.845 similarity. One part of isolates manifested the differentiation because of different origin and host. The pathogen of Cedrus deodara canker was proved to be B.dothidea according to its DNA polymorphism. So, the host of the pathogen was very wide, including angiosperm and gymnosperm. Two primers of RAPD analysis, K16 and R14, could be the identification primers as differentiating the isolates of Shanxi province and Hunan province from others. RAPD analysis demonstrated that the traditional taxonomy of B.berengeriana de Not.f.sp.piricola (Nose)Koganezawa et Sakuma as the specialization of B.berengeriana de Not. Was reasonable. And these two pathogens causing apple canker were different species from that one causing Poplar canker.

Key words: Botryosphaeria, 28S rDNA-PCR-FLP, RAPD, Identifying primers