摘要
应用变性梯度凝胶电泳(DGGE)技术以及主成分分析方法,结合植物病原菌传统分离、鉴定及致病力测定,检测位于黑龙江省海伦市大豆连作定位试验区根腐病原镰孢菌的种群构成及其致病力。采用真菌传统分离和鉴定方法共确定镰孢菌6个种,包括燕麦镰孢Fusarium avenaceum、木贼镰孢F.equiseti、禾谷镰孢F.graminearum、腐皮镰孢F.solani、尖镰孢F.oxysporum和拟轮枝镰孢F.verticillioides。其中,尖镰孢菌的分离频率最高,为56.7%。结合DGGE条带克隆测序及系统发育分析鉴定出8种镰孢菌,较传统方法增加了黄色镰孢F.culmorum和一个尖镰孢近似种。同时,DGGE图谱中尖镰孢所占百分比下降为37.4%。采用主成分分析方法将大豆生育指标相关的多种变量进行降维分析,得到第一主成分贡献值为91.2%。按照贡献值的正负结果,将30株镰孢菌分为致病和非致病类群。第二主成分贡献值为5.8%,根据其正负值可以矫正被错误估计的致病能力。综合所有研究结果,该定位试验区大豆根腐病主要病原镰孢菌为尖镰孢、禾谷镰孢和燕麦镰孢,且以尖镰孢菌为优势病原菌。
In this study,based on the results of conventional pathogen isolation,identification and pathogenicity test,the methods of denaturing gradient gel electrophoresis （DGGE） and principal component analysis （PCA） were applied to determine the population structure of pathogenic Fusarium species associated with soybean root rot in a continuous cropping soybean field located at the Hailun Experimental Station of Agricultural Ecology in Northeast China.After fungal isolation and morphological identification,six Fusarium species were determined,including F.avenaceum,F.equiseti,F.graminearum,F.solani,F.oxysporum and F.verticillioides.The maximum isolation ratio was 56.7％ of F.oxysporum.The DGGE fimgerprint offered 13 bands about tef-1 sequence of Fusarium,and eight bands had been cloned,sequenced and aligned successfully.Besides the above six Fusarium species determined by conventional isolation and identification,additionally the other two Fusarium species were identified,namely F.culmorum and F.oxysporum-like species.Meanwhile,the percentage of F.oxysporum decreased to 37.4％.According to principal component analysis,the first principal component （PC1） accounted for 91.2％ of variation in all data,which separated the Fusarium populations into pathogenic group and nonpathogenic group.The second principal component （PC2） weighed 5.8％ of all variation,which could adjust the error evaluation of pathogenicity.In conclusion,F.oxysporum,F.graminearum and F.avenaceum were confirmed as the pathogenic fungi of soybean root rot in this experimental station with F.oxysporum as the dominant pathogen.
出处
《植物病理学报》
CAS
CSCD
北大核心
2013年第5期500-508,共9页
Acta Phytopathologica Sinica
基金
中国科学院知识创新工程重要方向项目(KZCX2-YW-408-3)
