泡桐丛枝植原体河北平山和江西吉安株系胸苷酸激酶基因多态性分析

doi:10.11707/j.1001-7488.20140816

林业科学 ›› 2014, Vol. 50 ›› Issue (8): 108-118.doi: 10.11707/j.1001-7488.20140816

泡桐丛枝植原体河北平山和江西吉安株系胸苷酸激酶基因多态性分析

宋传生¹, 胡佳续², 林彩丽¹, 任争光¹, 耿显胜³, 田国忠¹

1. 中国林业科学研究院森林生态环境与保护研究所国家林业局森林保护学重点实验室北京 100091;
2. 天津出入境检验检疫局天津 300457;
3. 中国林业科学研究院亚热带林业研究所富阳 311400

收稿日期:2014-02-17 修回日期:2014-04-07 出版日期:2014-08-25 发布日期:2014-07-31
基金资助:
国家自然科学基金项目（30872025；31170602）。

Thymidylate Kinase Gene Polymorphism of Two Strains of Paulownia Witches’-Broom Phytoplasma

Song Chuansheng¹, Hu Jiaxu², Lin Caili¹, Ren Zhengguang¹, Geng Xiansheng³, Tian Guozhong¹

1. Key Laboratory of Forest Protection of State Forestry Administration Research Institute of Forest Ecology, Environment and Protection, CAF Beijing 100091;
2. Tianjin Entry-Exit Inspection and Quarantine Bureau Tianjin 300457;
3. Research Institute of Subtropical Forestry, CAF Fuyang 311400

Received:2014-02-17 Revised:2014-04-07 Online:2014-08-25 Published:2014-07-31
Contact: 田国忠

摘要/Abstract

摘要：

设计引物并PCR扩增河北平山株系PaWBPs和江西吉安株系PaWBJan的tmk基因，直接和克隆测定tmk序列，利用DNAMAN，MEGA等软件进行序列多样性、序列变异、蛋白功能域、系统进化等比较分析。结果表明：从PaWBPs和PaWBJan中克隆测序的93条和41条tmk-a中，分别有52和24种不同的序列，tmk-a ORF可被划分为2类，tmk-a -1 为639 bp，tmk-a -2 为627 bp，PaWBPs株系tmk-a -1 与tmk-a -2 序列条数的比值约为2.5，而PaWBJan株系为3.3；PaWBPs有5个相同的tmk-a -1 序列与PaWBJan的一个tmk-a -1 序列及枣疯植原体tmk-Y序列完全一致。因多位点突变，PaWBPs含假基因比例达48.1%，PaWBJan的为41.7%；tmk-b基因为单一序列，两株系的tmk-b核酸序列相似性为99.8%，编码蛋白仅有一个氨基酸差异。TMK-a和TMK-b中都具有胸苷酸激酶的3个保守功能域。系统进化分析显示，泡桐丛枝植原体的2个株系的所有tmk-a序列都与枣疯植原体tmk-X和tmk-Y等聚为一个进化枝Ⅰ中；而tmk-b与小麦蓝矮tmk -2 等聚为另一进化枝Ⅲ，基于tmk-b序列和基于16S rDNA序列构建的系统进化树一致，tmk-b有助于植原体16Sr组水平的分类，而tmk-a可用于植原体株系变异和遗传多样性分析。

关键词: 泡桐丛枝植原体, 胸苷酸激酶基因, 假基因, 功能域, 变异, 系统进化

Abstract:

The diversity, variation, predicted protein function domains and evolution of thymidylate kinase gene (tmk),which was obtained by cloning DNA sequencing of PCR products with designed primer from paulownia witches'-broom phytoplasmas Pingshan strain (PaWBPs) in Hebei Province and Ji'an strain (PaWBJan) in Jiangxi Province were comparatively analyzed by DNAMAN and MEGA software. The 52 and 24 different tmk-a homologues respectively from the cloned 93 sequences of PaWBPs and 41 of PaWBJan in total were identified, whereas the tmk-b gene sequence was relatively conserved, with 99.8% similarity and only one amino acid variation of the predicted proteins between the two strains. All tmk-a ORFs were classified into tmk-a-1 (639 bp) and tmk-a-2 (627 bp). The ratio of tmk-a-1/tmk-a-2 in PaWBPs and PaWBJan was 2.5 and 3.3, respectively. Five cloned tmk-a-1 sequences of PaWBPs were the same as one in PaWBJan as well as tmk-Y of jujube witches' broom phytoplasma. In addition, PaWBPs and PaWBJan respectivety contained up to 48.1% and 41.7% of predicted tmk-a pesudogenes in total tmk-a due to the multiple locus mutation. Rich sequence variability was found in tmk-a homologues of PaWBPs and PaWBJan and there existed three functional domains in amino acid sequences of both tmk-a and b which are necessary for thymidylate kinase activity. The phylogenetic analysis showed that all the tmk-a homologues of both strains with tmk-X and Y were clustered into clade I, while tmk-b with wheat blue dwarf tmk-2 into clade Ⅲ, suggesting that tmk-b nucleotide sequences consistent with highly conserved 16S rDNA could be used for the classification of phytoplasmas on 16Sr group levels whereas diverse tmk-a might be helpful for analyzing the genetic variation and diversity of different phytoplasma strains.

Key words: paulownia witches’-broom phytoplasma, thymidylate kinase gene, pesudogene, functional domain, variation, phylogenesis

中图分类号:

S763.14

宋传生, 胡佳续, 林彩丽, 任争光, 耿显胜, 田国忠. 泡桐丛枝植原体河北平山和江西吉安株系胸苷酸激酶基因多态性分析[J]. 林业科学, 2014, 50(8): 108-118.

Song Chuansheng, Hu Jiaxu, Lin Caili, Ren Zhengguang, Geng Xiansheng, Tian Guozhong. Thymidylate Kinase Gene Polymorphism of Two Strains of Paulownia Witches’-Broom Phytoplasma[J]. Scientia Silvae Sinicae, 2014, 50(8): 108-118.

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泡桐丛枝植原体河北平山和江西吉安株系胸苷酸激酶基因多态性分析

Thymidylate Kinase Gene Polymorphism of Two Strains of Paulownia Witches’-Broom Phytoplasma

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