摘要
为了研究番茄LCYE基因对其果实中番茄红素含量的影响机制,培育高品质番茄品种,在Gen Bank中根据已知的LCYE(ID:544 129)扩增出337 bp的保守序列;根据KJ 561 284.1扩增出2 203 bp的果实特异性启动子E8;查找番茄LCYE的内含子,选择第三个长112 bp的Intron片段;利用同尾酶构建出载体p Ri E8-LCYE,与p CAMBIA1301连接构建出果实特异性RNAi表达载体p CRi E8-LCYE,测序表明载体构建正确。农杆菌介导,叶盘法转化番茄,PCR检测表明得到了转基因番茄植株。与同样条件下番茄LCYB基因干扰载体的遗传转化过程相比,表明:对LCYE基因的干扰使番茄子叶开始褐化时间提前,且褐化率更高,说明筛选培养时子叶的褐化与所干扰的基因也有关系。
In order to study the influencing mechanism of LCYE gene on Lycopene content in tomato fruit and breed high-quality tomato varieties, firstly we amplified 337 bp conserved sequence according to the known LCYE(Genbank ID: 544 129), and amplified 2 203 bp fruit-specific promoter E8 according to KJ 561 284.1. Secondly seeking the LCYE intron in tomato and choosing the third length 112 bp. The vector p Ri E8-LCYE was constructed through isocaudarner to insert into p CAMBIA 1 301 and achieved the fruit-specific RNAi expression vector p CRi E8-LCYE. Sequencing results showed the vector was constructed correctly. The plasmid was transformed into tomato through leaf disc method mediated by Agrobacterium tumefaciems, and the PCR results showed the transgenic tomato plants were obtained. Compared with the transformation of tomato LCYB gene under the same conditions, the results showed: the interference of LCYE gene made tomato cotyledons browning bring forward and the browning rate higher, which indicates the transformation rate could be related to the gene interfered.
出处
《分子植物育种》
CAS
CSCD
北大核心
2016年第9期2371-2376,共6页
Molecular Plant Breeding
基金
农业部转基因生物新品种培育科技重大专项(2011ZX08005-002)资助
