谷子粒黑穗病菌PCR检测体系的建立  

Establishment of PCR Detection System for Ustilago crameri of Millet

作　　者：郝雅萍 焦思薇 郭二虎 仪慧兰[2] HAO Yaping;JIAO Siwei;GUO Erhu;YI Huilan(College of Agriculture,Shanxi Agricultural University,Taiyuan 030031,China;College of Life Science,Shanxi University,Taiyuan 030006,China;Institute of Millet,Shanxi Agricultural University,Changzhi 046000,China)

机构地区：[1]山西农业大学农学院,山西太原030031 [2]山西大学生命科学学院,山西太原030006 [3]山西农业大学谷子研究所,山西长治046000

出　　处：《山西农业科学》2021年第9期1031-1034,共4页Journal of Shanxi Agricultural Sciences

基　　金：国家自然科学基金项目(31972132);山西省重点研发计划(国际科技合作)项目(201903D421062);国家现代农业产业技术体系建设专项(CARS-06-13.5-A10)。

摘　　要：为了建立一种快速、有效鉴定谷子粒黑穗病菌(Ustilago crameri)的分子检测方法,使谷子粒黑穗病得到有效的监测和防治,采用真菌通用引物ITS1/ITS4对谷子粒黑穗病菌ITS序列进行扩增,扩增产物测序后,依据粒黑穗病菌序列设计合成了1对引物,分别以谷子粒黑穗病菌DNA、谷子白发病菌DNA、谷瘟病菌DNA、玉米黑粉菌DNA及谷子叶片基因组DNA作为模板进行PCR扩增。结果表明,仅谷子粒黑穗病菌DNA作模板时有1条特异性扩增条带,且检测的灵敏度可达10 pg/μL,其他材料均未出现扩增条带;将建立的PCR技术用于田间植株检测,能特异性检出谷子穗梗及旗叶中谷子粒黑穗病菌的存在;不同品种谷子植株中的病菌检出率与田间发病率数据基本一致,说明所用引物和PCR检测体系在鉴定粒黑穗病菌感染和植株抗病性方面具有较高的特异性。研究建立的谷子粒黑穗病菌PCR检测体系具有快速、准确、特异性和实用性强等特点,可为谷子粒黑穗病的监测和有效防治提供技术支持。To effectively monitor and control the Ustilago cramer-caused disease,a rapid and sensitive molecular detection method was required.In this study,the fungal universal primers ITS1/ITS4 were used to amplify the ITS sequence of Ustilago crameri.After obtained the sequence of above amplified product,a specific primer pair was designed and synthesized,and the genomic DNA molecular from Ustilago crameri spore,Sclerospora graminicola,Pyricularia setariae,Ustilago maydis and millet leaves were used as the template,respectively,to amplify a specific sequence of Ustilago crameri.The results showed that a specific band was observed only when the DNA template came from Ustilago crameri spore,while the sensitivity of the primer set for the detection of genomic DNA of Ustilago crameri was reached at 10 pg/μL,and no amplification product was found for other templates.Using above established PCR process to detect Ustilago crameri in field millet plants,the detectable rate of Ustilago crameri genomic DNA from foxtail millet plant peduncle and flag leaves was consistent with the data of its disease incidence rate in several different varieties.The present results indicated that the primer together with PCR process had a higher efficiency in identification of the smut infection and disease resistance to Ustilago crameri in plants.The PCR detection system established in this study was rapid,accurate,specific and practical,which could provide technical support for the monitoring and effective control of Ustilago crameri-caused disease.

关 键 词：谷子 粒黑穗病菌 PCR检测 

分 类 号：S515[农业科学—作物学]