番茄NAM基因的克隆与遗传转化

摘要

To understand the function of NAM gene in tomato, the full length cDNA NAM gene was amplified by RT-PCR from tomato cDNA, which contains a 1 053 bp open reading frame (ORF)encoding 351 amino acid residues, and contains NAC domain in its N terminus. Quantitative PCR analysis showed that NAM expressed highly in all tissues including root, stem, leaf, flower and fruit. To further investigate the function of tomato NAM gene, NAM overexpression vector pLP ?5S -NAM was constructed, and transformed into tomato plants by Agrobacterium-mediated method. The NAM overexpressing plants showed a dwarf phenotype, the size of these plants was smaller than those of wild-type plants, the plant growth was inhibited, and the leaves of transgenic tomato were highly lobed shaped. In conclusion, NAM affected the formation of shoot apical meristem(SAM)and leaf morphogenesis in tomato.%为了研究番茄NAM基因的功能,利用RT-PCR技术从番茄cDNA中分离了NAM(GenBank登录号:FJ435163.1)基因,该基因ORF全长1 053 bp,共编码351个氨基酸,N端含有NAC结构域.定量RT-PCR分析结果表明,该基因在番茄的根、茎、叶、花和果实中均有较强表达.为了进一步研究番茄NAM基因的功能,构建了NAM基因的超表达载体pLP100-35S-NAM,通过农杆菌介导法转化番茄,并获得抗性植株.经表型分析,转基因番茄植株矮小,生长受到抑制,叶片形态异常,表明NAM基因影响番茄顶端分生组织(SAM)的形成和叶片形态的发生.