狗蔷薇RcKN1基因的克隆及功能分析  

Cloning and Functional Characterization of RcKN1 Gene from Rosa canina L.

作　　者：姜福星[1] 张建甫[1] 刘凤栾[1] 马男[1] 赵梁军[1] 

机构地区：[1]中国农业大学农学与生物技术学院观赏园艺与园林系,北京100193

出　　处：《农业生物技术学报》2012年第7期715-725,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.30871733)

摘　　要：Class-Ⅰ KNOX(KNOX1)基因是真核生物中高度保守的同源盒转录因子,在植物的器官和胚胎发生中起着重要作用。本研究从狗蔷薇(Rosa canina L.)的类原球茎中克隆出了其同源基因,通过对其编码的蛋白质序列比对和系统发育分析,认为其属于KNOX1 在狗蔷薇中的同源基因;该基因全长 1 509bp,其中编码区 1 185 bp,编码 395 个氨基酸,将其命名为 RcKN1 基因 (GenBank 登录号:JN998201)。RT-PCR分析表明,其在叶芽、花芽、类原球茎等幼嫩的器官中表达;进一步构建了组成型表达载体并导入烟草(Nicotiana tabacum L.)中分析其功能,转基因烟草出现了叶片浅裂、叶脉紊乱、叶片基部歪斜、叶片皱缩甚至类似于复叶的叶片等多种表型。研究结果表明,RcKN1 基因在植物发育中起着重要作用,是研究叶片发育机理和培育叶型奇特的花卉新品种有价值的目的基因。Class- I KNOX （KNOX1） gene is highly conserved homeobox transcription factor in eukaryote, and play a vital role in the process of plant embryogenesis and organ development. Homologue gene of KNOX1 gene was clonded from protocorm-like body of Rosa canina L., through comparison amino acid sequences with other KNOXI gene and phylogenetic analysis, it was named RcKN1 gene. The full length cDNA of RcKN1 was 1 509 bp, with open reading frame （ORF） of 1 185 bp, encoding a protein of 395 amino acids, 5＇UTRs of 32 bp and 3＇UTRs of 291 bp （GenBank accession No. JN998201）. RT-PCR results showed that the gene was only expressed in young organ including leaf bud, floral bud, protocorm-like body, and so on. Constitutive plant expression vector of RcKN1 gene was constructed, its fimction was analysed by introducing it into tobacco （Nicotiana tabacum L.）. A series of novel phenotype were observed on transgenic tobacco, including lobbed leaf, disorder leaf vein, deficit leaves, even leaf like compound leaf, and so on. The results suggest RcKN1 gene performed important function during plant development, which is valuable target gene can be used to study mechanism of leaf development and breed new flower varieties with novel leaf type.

关 键 词：RcKN1基因 狗蔷薇的类原球茎 组成型表达 叶片形态 维管束发育 

分 类 号：S685.12[农业科学—观赏园艺]