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中国农学通报 ›› 2013, Vol. 29 ›› Issue (36): 74-79.doi: 10.11924/j.issn.1000-6850.2013-0568

所属专题: 生物技术 小麦

• 农学 农业基础科学 • 上一篇    下一篇

小麦三雌蕊品系UCR–14KD基因cDNA序列克隆及表达分析

蒋泽元 彭正松 杨在君 吴凯 欧阳钟鸣 巫有霞   

  • 收稿日期:2013-03-04 修回日期:2013-03-30 出版日期:2013-12-25 发布日期:2013-12-25
  • 基金资助:
    国家自然科学基金项目

Sequence Cloning and Expression Analysis of UCR-14KD from the Common Wheat Line with Three Pistil

  • Received:2013-03-04 Revised:2013-03-30 Online:2013-12-25 Published:2013-12-25

摘要: 小麦UCR-14KD基因是呼吸链中众多重要基因之一,其表达量影响小麦呼吸速率和能量代谢,进而对小麦雌蕊和雄蕊的发育起着重要的调控作用。为进一步了解小麦UCR-14KD基因的结构特点及其在小麦三雌蕊突变体(TP)形成的分子机制中所发挥的作用,本文利用RT-PCR技术从三雌蕊突变体近等基因系CSTP幼穗中克隆了TPUCR-14KD基因完整的编码区序列,并将其转入大肠杆菌进行了诱导表达,对其cDNA及推导的氨基酸序列进行分析,结果显示:该基因编码区长381bp, 编码126个氨基酸,二级结构为α螺旋和无规卷曲,通过实时定量PCR检测,显示该基因在CSTP幼穗中大量表达,而在幼穗不同发育阶段的表达量为:幼穗长度2-5mm﹥5-7mm﹥7-10mm,并且显著高于该基因在CS幼穗对应阶段的表达量。这表明TPUCR-14KD基因可能在三粒小麦形成的过程中,尤其是在其雌蕊原基分化发育初期起到了重要作用。本研究结果对于进一步明确TPUCR-14KD基因的结构和功能,尤其是TPUCR-14KD在小麦三雌蕊形态构建中的作用奠定了基础。

关键词: 毛乌素沙地, 毛乌素沙地

Abstract: Three pistils is a mutation in common wheat,it could potentionally increase the number of gains per spike, so it is a valuable mutant for wheat breeding. To study the molecular mechanism of the three pistils mutation in wheat we cloned and characterized the cDNA sequence of TPUCR. Then, the acquired sequence and the amino acids were analyzed and their sequences alignment, secondary structure and functional domains were predicted. Moreover, the TPURD gene was over-expressed in E. coli. The expression patterns of TPURDwere analyzed through real-time quantitative RT-PCR in different developmental stages of young spikes. The open reading frame (ORF) of TPUCR was 381 bp, and it encodes 126 amino acids. Alignment analysis indicates that the deduced amino acid sequence share a high homology with those of other several grain crops. Functional domains analysis indicated that the TPURD contained a conserved domain. Expression analysis indicates that there are higher levels of transcripts in the earlier developmental stages of young spikes. The transcript levels in the young spikes of Chinese Spring three pistils line increased by 4-fold compared with that in the young spikes of the Chinese Spring line. The data provide a basis for further research on function of TPURD, and a rationale for the molecular mechanism of the three pistil mutation in wheat

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