| 黎飞飞，李 波，乔 梦，闵东红，张小红.小麦愈伤组织再生相关基因 TaTCP 1A的克隆及功能鉴定[J].麦类作物学报,2014,34(7):885 |
| 小麦愈伤组织再生相关基因 TaTCP 1A的克隆及功能鉴定 |
| Cloning and Characterization of TaTCP 1A Gene Relevant to Plant Regeneration in Wheat |
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| DOI：10.7606/j.issn.1009-1041.2014.07.02 |
| 中文关键词: 小麦 愈伤组织 TaTCP 1A 遗传转化 RNAi qRT PCR |
| 英文关键词:Wheat Callus TaTCP 1A Genetic transformation RNA interference qRT PCR |
| 基金项目:国家转基因重大专项（2014ZX08002 003B）；陕西省农业科技攻关项目（2013K02 01） |
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| 中文摘要: |
| 为深入探讨小麦再生分子机制、给建立高效的小麦遗传转化受体系统奠定基础,通过电子拼接、RT PCR及RACE等方法在小麦品种郑麦7698中分离到一个与小麦愈伤组织再生相关的候选基因 TaTCP 1A。序列分析表明，该基因的cDNA序列全长为2 163 bp,其中包含了一个1 623 bp的开放阅读框，编码540个氨基酸。qRT PCR结果表明，该基因具有组织表达特异性，在愈伤组织中表达量最高；在愈伤组织形成阶段，随诱导时间的延长，其表达量逐渐上升，当诱导11 d时表达量达到峰值，随后呈下降趋势；此外， TaTCP 1A基因在胚性愈伤组织中的表达量高于非胚性愈伤组织。基因沉默结果表明，转 TaTCP 1A RNAi小麦植株的再生率比对照降低了85.09%，这一结果初步证明该基因对小麦愈伤组织再生有一定的促进作用。 |
| 英文摘要: |
| Good receptor with high plant regeneration ability is a key factor for the genetic transformation of wheat. Therefore, understanding the molecular mechanism of plant regeneration in vitro is the core of establishment of efficient wheat genetic transformation system. A candidate gene relevant to wheat regeneration, TaTCP 1A, was isolated from wheat cultivar Zhengmai7698 through electronic splicing, RT PCR, and RACE. Sequence analysis indicated that the cDNA of this gene contained 2 163 nucleotides in length, with a 1 623 bp complete open reading frame encoding 540 amino acids. The qRT PCR results showed that TaTCP 1A was highly expressed in callus; its expression gradually rised with the extension of callus induction time, and reached a peak at 11d after induction, and then showed a downward trend; the transcription level of TaTCP 1A in embryonic callus were higher than in non embryonic callus. Preliminary function analysis of T1 transformed wheat with TaTCP 1A RNAi cassette showed negative effect on regeneration efficiency, which indicated that the gene may play a role in promoting regeneration. |
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