光皮桦BlSPL1转录因子基因的克隆、表达及单核苷酸多态性分析

摘要/Abstract

摘要：

Squamosa promoter binding protein like genes(SPLs)对植物开花具有重要的调控作用。基于转录组序列并利用RACE技术从光皮桦中分离获得BlSPL1基因,其cDNA序列全长1825bp,包含开放阅读框长1170bp,编码389个氨基酸,与桃PpSPL1(EMJ10405)的同源性最高(67%),属于陆地植物SPLs基因家族的groupVII分支。表达分析显示BlSPL1在光皮桦芽、雌花、雄花、幼苗中均有表达,在雄花和幼苗中表达量较低,但在芽发育成雌花过程中明显上调,推测其可能对早期的雌花形成起调控作用。同时,从57个不同的光皮桦无性系中克隆BlSPL1的基因组序列,进行单核苷酸多态性(SNP)分析,共检测到98个SNP位点,SNP频率为1/26bp,多样性指数π为0.00712。在外显子区域,共检测到28个SNP位点,其中11个为同义突变,17个为错义突变。进一步的连锁不平衡分析显示,随着序列长度的增加,不同光皮桦群体BlSPL1内的SNPs连锁不平衡在基因内部迅速衰退。

关键词: 光皮桦, 转录因子, SPL基因, 单核苷酸多态性

Abstract:

Squamosa promoter binding protein like genes(SPLs)play an important role in regulating flowering of plants. In this study, one SPL gene named BlSPL 1 was cloned from Betula luminifera using RACE (rapid-amplification of cDNA ends) strategy. The cDNA of BlSPL 1 is 1 825 bp in length with an open reading frame (ORF) encoding a protein of 389 aa. The deduced protein sequence of the BlSPL 1 shares the highest identity with PpSPL 1 (EMJ10405) (67%) and belongs to group Ⅶ of SPL family from land plants. Expression analysis indicated that the BlSPL 1 transcripts were at a relatively low level in male flower and seedlings, but were apparently up-regulated during the development from buds to female flowers, which implied that BlSPL 1 might play important roles in early stage of female flower formation. At the same time, the corresponding genomic fragments of BlSPL 1 were also cloned from 57 different germplasms for the single nucleotide polymorphism (SNP) analysis. A total of 98 SNPs were detected, and the frequency and diversity of SNPs were 1/26 bp and 0.007 12, respectively. There were 28 SNPs detected in the exons of BlSPL 1, among which, there were 11 synonymous and 17 missense mutations SNPs, respectively. Linkage disequilibrium (LD) analysis showed that LD declined rapidly within the BlSPL 1 along with the length increasing in different population of B. luminifera, suggesting that LD mapping based on BlSPL 1 gene would be feasible and useful in B. luminifera.

Key words: Betula luminifera, transcription factor, SPL genes, single nucleotide polymorphism

中图分类号:

S718.46

李玉岭, 周厚君, 林二培, 黄华宏, 徐莉莉, 童再康. 光皮桦BlSPL1转录因子基因的克隆、表达及单核苷酸多态性分析[J]. 林业科学, 2013, 49(9): 52-61.

Li Yuling, Zhou Houjun, Lin Erpei, Huang Huahong, Xu Lili, Tong Zaikang. Isolation, Expression and Single Nucleotide Polymorphisms Analysis of Transcription Factor (BlSPL 1) from Betula luminifera[J]. Scientia Silvae Sinicae, 2013, 49(9): 52-61.

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