新疆农业科学2013，Vol.50Issue(3)：476-483,8.DOI:10.6048/j.issn.1001-4330.2013.03.014
新疆枣疯病植原体tuf基因的克隆与序列分析
Cloning and Sequencing Analysis of tuf Gene of the P hytoplasma Associated with Jujube Witches Broom in Xinjiang
摘要
Abstract
[Objective] Jujube witches broom ( JWB ) is an important plant disease caused by phytoplasma. The major objective of our research was to realize the molecular phylogenetic relationship, genetic differentiation and obtain molecular characteristics of JWB phytoplasmas of Xinjiang isolates. [ Method ] By using of PCR, the elongation factor Tu ( tuf gene) of phytoplasma associated with jujube witches broom in the total DNA extracted from diseased samples in Aksu and Kashi region of Xinjiang was amplified with the universal primer pairs fTufu/rTufu, then the partial tuf gene was cloned and sequenced, and similarity to other phytoplasmas was analyzed. [Result]The sequencing result revealed that tuf gene of phytoplasmas from JWB - ASZ strain isolated in Aksu was 841 bp in length and JWB - KHZ strain isolated in Kashi was 814 bp in length. Homology analysis of sequences of tuf gene showed that the JWB - ASZ and JWB - KHZ were closely related to those in the 16S rV - B subgroup of jujube witches' broom phytoplasma. In tuf gene, JWB - KHZ strain isolated in Kashi shared most similarity with JWB strain Hebei which had been already reported, and homology rates of nucleotide sequences and amino acids were 90. 2% and 81. 5% , respectively. JWB - ASZ strain isolated in Aksu shared most similarity with JWB in Shaanxi district, and homology rates of nucleotide sequences and amino acids were 94. 2% and 94. 6% , respectively. However, they showed a low similarity ( <70% ) with members of the other 16S rV subgroup phytoplasmas. And the remarkable differences of the tuf gene nucleotide and the derived amino acid sequence similarities between the two isolates were 73. 5% and 63.5 %. [ Conclusion ] This paper for the first time reports the tuf gene sequences of JWB phytoplasmas of Xinjiang isolates, thus directly providing a theoretical basis for research of the phytoplasma molecular characterization, molecular variation and genetic diversity of the different phytoplasmas associated with JWB in Xinjiang.关键词
枣疯病/植原体/tuf基因/序列分析Key words
jujube witches'broom/ phytoplasma/ elongation factor Tu/ sequence analysis分类
农业科技引用本文复制引用
韩剑,徐金虹,王同仁,张祥林,罗明..新疆枣疯病植原体tuf基因的克隆与序列分析[J].新疆农业科学,2013,50(3):476-483,8.基金项目
新疆维吾尔自治区自然科学基金(2010211A17) （2010211A17）
