摘要
为确定贵州玛瑙红樱桃花变绿病害病原分类地位及构建快速有效的检测体系,开展了研究。利用植原体16S rRNA基因引物R16m F2/R16m R1对感病玛瑙红樱桃总DNA进行常规PCR扩增、克隆和测序,通过同源性比较、系统进化分析和在线虚拟RFLP分析,确定其属于植原体16SrV-B亚组。根据16S rRNA基因设计并合成引物对MN16SF-2/MN16SR-2,用重组质粒pEASY-T1-MN16S为模板建立标准曲线,构建了玛瑙红樱桃花变绿植原体实时荧光定量PCR检测体系。运用该体系检测喷施了外源茉莉酸甲酯、褪黑素、四环素和未施外源物质的患病苗木体内植原体含量,发现施加外源物质组体内植原体含量与未施组相比均大幅下降。研究明确了玛瑙红樱桃花变绿植原体的分类地位并成功建立了qPCR检测体系,为玛瑙红樱桃植原体检测提供了技术支持。
In order to determine the taxonomic status of the phytoplasma causing virescence disease in Prunus pseudocerasus‘Manaohong’in Guizhou and to construct a rapid and effective detection system,the phytoplasma 16S rRNA gene primers R16m F2/R16m R1 were used for PCR amplification,clone and sequence of the total DNA of diseased P.pseudocerasus‘Manaohong’.It was determined that identified phytoplasma belonged to the phytoplasma 16SrV-B subgroup through homology comparison,phylogenetic analysis and online virtual RFLP analysis.According to the 16S rRNA gene,primer pair of MN16SF-2/MN16SR-2 was designed and synthesized.The standard curve was established with the recombinant plasmid pEASY-T1-MN16S as template,and a real-time fluorescence quantitative PCR detection system for P.pseudocerasus‘Manaohong’virescence phytoplasma was constructed.By applying this detection system to monitor phytoplasma content of diseased seedlings sprayed with exogenous methyl jasmonate(MeJA),melatonin,tetracycline,and non-treated controls.It was found that phytoplasma contents in the treatment groups were significantly reduced compared to the control.In this study,the taxonomic status of P.pseudocerasus‘Manaohong’virescence phytoplasma was clarified,and a qPCR detection system was successfully established,which provided technical support for the detection of P.pseudocerasus‘Manaohong’phytoplasma.
作者
何梅
朱淑珠
王雅丽
王林
李燏
宋常美
HE Mei;ZHU Shuzhu;WANG Yali;WANG Lin;LI Yu;SONG Changmei(School of Biology and Environmental Engineering,Guiyang University,Guiyang,Guizhou,550005,China)
出处
《中国南方果树》
北大核心
2025年第3期125-131,共7页
South China Fruits
基金
国家自然科学基金地区基金(32160693)
贵州省林业科学技术研究项目(黔林科合[2020])
贵阳学院学科团队建设项目(合同号2022-xk12)资助。
关键词
玛瑙红樱桃
花变绿
病原鉴定
实时荧光PCR检测
Prunus pseudocerasus‘Manaohong’
virescence
pathogen identification
real-time fluorescence PCR detection
