安徽桑黄花型萎缩病植原体16S rDNA序列分析及分子检测  

Molecular detection and sequence analysis on 16S ribosomal DNA of phytoplasma associated with mulberry yellow dwarf disease occurring in Anhui

作　　者：卢全有[1,2] 

机构地区：[1]中国农业科学院蚕业研究所农业部蚕桑遗传改良重点开放实验室,镇江212018 [2]江苏科技大学,镇江212003

出　　处：《植物病理学报》2010年第6期642-646,共5页Acta Phytopathologica Sinica

基　　金：江苏省自然科学基金项目(BK2006084)

摘　　要：Mulberry yellow dwarf(MYD)disease is an quarantine disease and the causal agent is a phytoplasma.Two pairs of published universal primer,P1/P7 and Rm16F2/Rm16R1,based on the 16S-23S rDNA sequence of phytoplasma and total DNA extracted from infected mulberry tissues were employed for PCR and nested-PCR detection.The results revealed that a phytoplasma-specific 1 830 bp fragment with a G+C content of 46.01% was sequenced(GenBank accession No.GQ249410).The sequence shared 99.7% and 99.8% identity with aster yellows,the representatiive phytoplasma in 16SrI group,and mulberry dwarf phytoplasma classified into subgroup B in 16SrI group and named as the MYD phytoplasma strain Anhui(MYD-Anh).A phylogenetic tree based on 16S rDNA sequences was constructed and showed that MYD-Anh was clustered into 16SrI group.Identity of 16S rDNA sequence between MYD-Anh and mulberry yellow dwarf phytoplasma strain Zhenjiang(MD-zj) was nearly 100%,and they might belong to the same strain.Nested-PCR was used to detect the pathogenic phytoplasma from the differential tissues of mulberry infected with MYD-Anh.The results showed that a phytoplasma-specific 1.4 kb fragment was amplified with total DNA extracted from bark and vein.Nested-PCR was more sensitive than PCR for detecting MYD phytoplasma.Mulberry yellow dwarf（MYD）disease is an quarantine disease and the causal agent is a phytoplasma.Two pairs of published universal primer,P1/P7 and Rm16F2/Rm16R1,based on the 16S-23S rDNA sequence of phytoplasma and total DNA extracted from infected mulberry tissues were employed for PCR and nested-PCR detection.The results revealed that a phytoplasma-specific 1 830 bp fragment with a G＋C content of 46.01% was sequenced（GenBank accession No.GQ249410）.The sequence shared 99.7% and 99.8% identity with aster yellows,the representatiive phytoplasma in 16SrI group,and mulberry dwarf phytoplasma classified into subgroup B in 16SrI group and named as the MYD phytoplasma strain Anhui（MYD-Anh）.A phylogenetic tree based on 16S rDNA sequences was constructed and showed that MYD-Anh was clustered into 16SrI group.Identity of 16S rDNA sequence between MYD-Anh and mulberry yellow dwarf phytoplasma strain Zhenjiang（MD-zj） was nearly 100%,and they might belong to the same strain.Nested-PCR was used to detect the pathogenic phytoplasma from the differential tissues of mulberry infected with MYD-Anh.The results showed that a phytoplasma-specific 1.4 kb fragment was amplified with total DNA extracted from bark and vein.Nested-PCR was more sensitive than PCR for detecting MYD phytoplasma.

关 键 词：RDNA序列分析 萎缩病 植原体 桑黄 分子检测 花型 16S 检疫性病害 

分 类 号：S888.712[农业科学—特种经济动物饲养]