贵州主栽樱桃品种的植原体分子检测及鉴定  

Molecular Detection and Identification of Phytoplasma of Cherry Cultivars in Guizhou Province

作　　者：田文杰 乔光 文晓鹏 田田 洪怡 TIAN Wenjie;QIAO Guang;WEN Xiaopeng;TIAN Tian;HONG Yi(Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Regions(Ministry of Education),Collaborative Innovation Center for Mountain Ecology ArgoBioengineering(CICMEAB),Institute of Argo-Bioengineering/College of Life Sciences,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学生命科学学院/农业生物工程研究院,山地植物资源保护与种质创新教育部重点实验室,山地生态与农业生物工程协同创新中心,贵阳550025

出　　处：《种子》2022年第2期18-24,共7页Seed

基　　金：贵州省科技厅项目“贵州樱桃植原体病害遗传多样性研究及无病苗培育”(黔科合支撑[2020]1 Y 133号)。

摘　　要：为确定贵州主栽樱桃品种玛瑙红、黑珍珠发生的花变叶、丛枝现象的病原分布、种类及其分类地位,本研究优化植原体16 S rDNA基因通用引物对R 16 mF 2/R 16 mR 2和R 16 F 2 n/R 16 R 2的巢氏PCR反应体系,同时对86个樱桃样品总DNA进行巢式PCR扩增,对扩增PCR产物进行克隆、测序及序列分析。结果表明,纳雍县、福泉市、乌当区等地样品植原体检出率高达100%;威宁县、沿河县、大方县分别为40.74%、66.67%和40%;序列同源性比较表明,获得的GZ-1、GZ-2植原体序列属于16 Sr V组;系统进化树及虚拟RFLP分析显示,GZ-1植原体序列属于16 Sr V-B亚组成员,GZ-2序列为16 Sr V组新的亚组。本研究确定了贵州主栽樱桃品种的植原体病害侵染情况,明确了其分类地位,为该病害检测和防控提供一定理论依据。To ascertain the occurrence of flower leaf change on mainly cultivated cherry varieties Manaohong and Heizhenzhu in Guizhou province and distribution,species and taxonomic status of pathogens of witch's-broom.In this study,we optimized the nested PCR reaction system of primer pair of R 16 mF 2/R 16 mR 2 and R 16 F 2 N/R 16 R 2 used to the universal primers of phytoplasma 16 S rDNA,and the total DNA of 86 cherry samples was amplified by nested PCR,and the PCR products were cloned,sequenced and analyzed further.The results showed that the phytoplasma detection rate in Nayong county,Fuquan city and Wudang district was as high as 100%,while detection rate in Weining county,Yanhe county and Dafang county were 40.74%,66.67%and 40%,respectively.Sequence homology comparison showed that the sequences of phytoplasma GZ-1 and GZ-2 belonged to 16 Sr V group.Phylogenetic tree and virtual RFLP analysis showed that GZ-1 was a member of 16 Sr V-B subgroup,and GZ-2 was a new subgroup of 16 Sr V group.In this study,the phytoplasma infestation of main cherry varieties in Guizhou province was determined,and its classification status was also clarified,providing a theoretical basis for the detection and prevention and control of the disease.

关 键 词：贵州樱桃 植原体 巢式PCR 16 S rDNA RFLP分析 

分 类 号：S662.5[农业科学—果树学]