高粱病程相关蛋白基因的鉴定及荧光定量PCR引物筛选  

Identification of Pathogenesis-related Protein Genes and Screening of Fluorescent Quantitative PCR Primers in Sorghum

作　　者：邵玉涛 刘丹 郭磊 王振杰 裴海霞 Shao Yutao;Liu Dan;Guo Lei;Wang Zhenjie;Pei Haixia(School of Life Science and Technology,Inner Mongolia University of Science and Technology,Baotou,014010)

机构地区：[1]内蒙古科技大学生命科学与技术学院,包头014010

出　　处：《分子植物育种》2020年第19期6392-6398,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31660396);内蒙古自然科学基金项目(2016MS0308);内蒙古科技大学创新基金优秀青年科学基金项目(2017YQL09);内蒙古科技大学大学生科技创新基金项目(2016017)和内蒙古科技大学创新基金(2016-QDL-B10)共同资助。

摘　　要：病程相关蛋白(pathogenesis-relatedproteins,PRs)在植物抗病反应中扮演着重要角色,然而,目前关于其在高粱抗蚜反应中是否发挥作用还不是很明确。为探究高粱抗蚜反应中病程相关蛋白的作用,需利用荧光定量PCR(fluorescent quantitative PCR,qPCR)的方法对该类基因的表达量变化进行检测,而PCR引物的筛选将直接影响到检测结果的准确性。本研究通过对蚜虫侵染高粱材料的转录组数据进行分析,筛选出17个病程相关蛋白基因,利用qPrimerDB数据库网站查找到17对引物。利用PCR和荧光定量PCR的方法,双重验证引物的特异性。最终筛选出6个基因的6对引物峰值单一,特异性和扩增效率均较为理想,可用于后续研究这些基因的表达模式变化,揭示病程相关蛋白在高粱抗蚜反应中的作用,为开展主要农作物抗虫分子育种提供理论依据。Pathogenesis-related proteins(PRs)play an important role in plant defense against pathogens,however,it is not clear whether they play a role in Sorghum resistance to aphids.In order to explore the role of PRs in Sorghum resistance to aphids,it is necessary to detect the expression of PR genes by fluorescence quantitative PCR(qPCR),and the selection of PCR primers will directly affect the accuracy ofdetection results.In this study,by analyzing the transcriptome data of Sorghum infected by aphids,17 PR genes were screened out,and 17 pairs of primers were found by qPrimerDB database.The specificity of the primers was verified by PCR and qPCR.Finally,six pairs of primers were screened out,with single peak value,good specificity and ideal amplification efficiency.These primers can be used to further study the expression pattern of these genes,reveal the role of PRs in aphid resistance in Sorghum,and provide theoretical basis for the development of insect-resistant crops.

关 键 词：高粱(Sorghum) 病程相关蛋白 荧光定量PCR 

分 类 号：S435.14[农业科学—农业昆虫与害虫防治] Q943.2[农业科学—植物保护]