农杆菌介导的P_(SAG12)-IPT基因转化柳枝稷研究  

Agrobacterium-mediated Transformation of P_(SAG12)-IPT Gene in Switchgrass

作　　者：柴乖强[1] 徐开杰[1] 王勇锋[1] 孙风丽[1] 刘曙东[1] 奚亚军[1] 

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100

出　　处：《草地学报》2014年第6期1331-1336,共6页Acta Agrestia Sinica

基　　金：国家自然科学基金(31171607)(31371690);国家948计划项目(2011-G1-19)资助

摘　　要：为延长生物能源作物柳枝稷(Panicum virgatum)的叶片功能期,以柳枝稷‘西稷1号’品系的成熟胚诱导的愈伤组织为受体,采用农杆菌介导法转化叶片衰老延缓基因PSAG12-IPT。结果表明:经过农杆菌转化,共培养,抗性愈伤组织筛选、再生和生根培养,在农杆菌介导处理的8500块愈伤组织中共获得了320株转化植株;经PCR检测分析,转化植株中共有19株植株扩增出标记基因潮霉素B磷酸转移酶基因的目标片段,初步表明PSAG12-IPT基因已整合到柳枝稷基因组中,转化率为0.223%。对柳枝稷转基因植株的叶片叶绿素含量、净光合速率、胞间CO2浓度和气孔导度进行测定,表明PSAG12-IPT基因在部分转基因植株中已经表达。In order to prolong the leaf functional period of bioenergy crop switchgrass,the leaf senescence gene PSAG12-IPT was transferred into switchgrass using Agrobacterium-mediated transformation with the embryogenic callus from mature seeds as the materials.The results showed that 320 transgenic plants were obtained from 8500 callus after agrobacterium-mediated transformation,cocultivation,screening resistant callus,plant regeneration and rooting.PCR analyses of transgenic plants indicated that the target fragment of Hygromycin B phosphoric acid transferase gene was amplified from 19 plants.It preliminary showed that the PsAG12-IPT gene was integrated into the genome of switchgrass,and the transformation efficiency was 0.223%.Further research on the leaf chlorophyll content,net photosynthetic rate,intercellular CO2 concentration and stomatal conductance of transgenic plants showed that PsAG12-IPT gene was expressed in some of the transgenic plants.

关 键 词：柳枝稷 PSAG12-IPT基因 遗传转化 农杆菌 

分 类 号：S36[农业科学—作物栽培与耕作技术]