小麦三雌蕊品系UCR–14KD基因cDNA序列克隆及表达分析

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。

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

To study the structure and the role of wheat UCR-14KD gene in the formation process of wheat three pistils trait. The open reading frame （ORF） sequence of TPUCR-14KD gene was cloned by RT-PCR technology from young spikes from near-isogenic lines of wheat three pistils CSTP. And then the expression patterns of this gene were analyzed using real-time PCR. The open reading frame （ORF） of TPUCR-14KD was 381 bp, and encoded 126 amino acids. The secondary structure of TPUCR-14KD constituted by c~-helix and random curls. Real-time PCR analysis showed that the TPUCR-14KD gene over-expressed in young spikes of CSTP. The expression levels of： the young spike length of 2 to 5 mm〉 5 to 7 mm〉 7 to 10 mm, and the expression level was significantly higher than the corresponding stage of the young spikes in CS. The TPUCR-ldKD gene may play an important role in the process of formation of three pistil trait in common wheat line three pistil （TP）.