南京林业大学学报(自然科学版) ›› 2013, Vol. 37 ›› Issue (03): 17-22.doi: 10.3969/j.issn.1000-2006.2013.03.004

• 研究论文 • 上一篇    下一篇

白桦SPL8转录因子基因的分离及转录表达分析

官民晓,刘雪梅*,张 妍,刘 瀛,孙丰宾   

  1. 东北林业大学生命科学学院,黑龙江 哈尔滨 150040
  • 出版日期:2013-06-18 发布日期:2013-06-18
  • 基金资助:
    收稿日期:2012-03-18 修回日期:2012-06-23
    基金项目:国家自然科学基金项目(31100449); 黑龙江省自然科学基金项目(C201040); 东北林业大学研究生论文资助项目(ST1P10)
    第一作者:官民晓,硕士生。*通信作者:刘雪梅,副教授,博士。liuxuemei@nefu.edu.cn。
    引文格式:官民晓,刘雪梅,张妍,等. 白桦SPL8转录因子基因的分离及转录表达分析[J]. 南京林业大学学报:自然科学版,2013,37(3):17-22.

Isolation and transcription expression analysis of SPL8 transcription factors gene of Betula platyphylla

GUAN Minxiao, LIU Xuemei*, ZHANG Yan, LIU Ying, SUN Fengbin   

  1. College of Life Science,Northeastern Forest University, Harbin 150040, China
  • Online:2013-06-18 Published:2013-06-18

摘要: 利用RACE和RT-PCR技术对白桦SPL转录因子SPL8进行克隆,获得cDNA全长为1 230 bp,其完整的开放读码框为930 bp,编码309个氨基酸,包含1个高度保守的SBP域,预测编码蛋白的分子质量为34.32 ku,理论等电点为9.08。将该基因命名为BplSPL8,并将它的cDNA序列提交到Genbank,登录号为JQ354964。同源序列比对分析表明,白桦BplSPL8与拟南芥AtSPL8、大豆GmSPL8、马铃薯ScSPL8、葡萄VvSPL8的氨基酸序列相似性分别为55.4%、73.1%、63.9%、70.8%。qRT-PCR结果显示,BplSPL8基因在不同组织均有表达,但在雄花序中最高,在种子和成熟花粉中最低; BplSPL8在雄花序不同发育阶段的表达量不同,另外,在雄花序发育突变体各组织中的表达水平均低于野生型。推测BplSPL8在白桦雄花序发育中的特定阶段起重要作用,其异常表达可能与雄性不育有关。

Abstract: A 1 230 bp full-length cDNA of SPL8 gene was isolated from Betula platyphylla Suk. using RACE(rapid amplification of cDNA ends)and RT-PCR(reverse transcription polymerase chain reaction). SPL8 gene contains an open reading frame of 930 bp which encodes 309 amino acid with the deduced molecular mass around 34.32 ku and the theoretical isoelectric point of 9.08.The gene was named BplSPL8 and submitted to GenBank with accession number JQ354964.The homology of amino acid compared with Arabidopsis thaliana AtSPL8, Glycine max GmSPL8, Solanum chacoense ScSPL8 and Vitis vinifera VvSPL8 was 55.4%,73.1%,63.9% and 70.8%, respectively. The qRT-PCR result showed that BplSPL8 is expressed in all different tissues, the expression was the highest in the male inflorescences, whereas lowest in the seed and pollen. The expression level changed at different development stages in the male inflorescences. In addition, the expression levels of BplSPL8 in mutant was below that in wild-type in all different tissues, which suggests that BplSPL8 may play an important role on the development’s specific stage of the male inflorescences in the Betula and its abnormal expression may be associated with male sterility.

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