摘要
根据玉米基因组数据MaizeGDB(http://www.maizegdb.org/)已经公布的玉米VHA-B(V-ATPase subunit B)基因的mRNA序列(AY104180),从干旱处理玉米材料CN165的花丝中利用RT-PCR技术获得VHA-B基因的编码序列,并进行了序列分析和Northern杂交分析。结果表明:VHA-B基因编码487个氨基酸,含有1个保守的ATP结合位点,其编码的蛋白分子量为54.09kD,等电点为4.99。氨基酸同源性分析表明:V-ATPase B亚基是一个较为保守的蛋白亚基。Northern杂交结果表明:在干旱处理下,B亚基存在两个不同的转录本,它们对干旱的响应不同;在盐胁迫下,同样也存在两个转录本,它们都对盐胁迫做出了响应;在冷处理下,仅存在1个转录本,且它的表达较强,仅仅在6h有一些微小地反复;在ABA处理下,也仅存在1个转录本,处理1h后表达明显上调,此后一直保持较高的表达水平。玉米VHA-B基因对花期干旱有响应,但在花丝和雌穗中的表达响应方式不同,在花丝中干旱诱导表达明显,而在雌穗中存在表达没有明显差异的两个转录本。
Based on mRNA sequence of VHA-B gene published in Maize GDB, the coding region sequence of ZmVHA -B was obtained by RT-PCR from maize inbred line CN165. Sequence analysis and Northern blotting were further carried out. The results indicated that this gene encoded a polypeptide of 487 amino acid residues, containing a conservative ATP binding site, with a molecular weight of 54.09 kD and an isoelectric point of 4.99. The analysis of amino acid homology indicated that V-ATPase B subunit was very conservative and it could be used in investigation of biological evolution. The results of the North blotting analysis showed that under drought stress at seedling stage, this gene existed in two transcripts which responded to drought differently. Under salt stress, this gene also existed in two transcripts with same expression pattern. Under cold stress and ABA treatment, this gene only existed in one transcript. Under cold stress its transcription level increased with some fluctuation. Under ABA treatment, the expression of VHA -B gene increased significantly from the time point of 1 h. after treatment, and kept steadily. VHA - B gene responded to drought at flowering stage of maize. However, the expression was different in silks and ears because induced expression of VHA -B gene with two transcripts was strong in the silks under drought while there were not significant changes of expression in the ears responding to drought.
出处
《玉米科学》
CAS
CSCD
北大核心
2009年第2期19-23,28,共6页
Journal of Maize Sciences
基金
国家“863”项目(2006AA10Z188)
国家自然科学基金(30571133)
科技部国际合作项目(0502)
