农杆菌介导的PSAG12-IPT基因转化柳枝稷研究

doi:10.11733/j.issn.1007-0435.2014.06.028

草地学报 ›› 2014, Vol. 22 ›› Issue (6): 1331-1336.DOI: 10.11733/j.issn.1007-0435.2014.06.028

农杆菌介导的P_SAG12-IPT基因转化柳枝稷研究

柴乖强, 徐开杰, 王勇锋, 孙风丽, 刘曙东, 奚亚军

西北农林科技大学农学院, 陕西杨凌 712100

收稿日期:2014-01-06 修回日期:2014-04-17 出版日期:2014-12-15 发布日期:2014-12-01
通讯作者: 奚亚军
作者简介:柴乖强(1986-),男,陕西扶风人,博士研究生,研究方向为农业生物技术与育种,E-mail:chaiguaiqiang@126.com
基金资助:
国家自然科学基金(31171607)(31371690); 国家948计划项目(2011-G1-19)资助

Agrobacterium-mediated Transformation of P_SAG12-IPT Gene in Switchgrass

CHAI Guai-qiang, XU Kai-jie, WANG Yong-feng, SUN Feng-li, LIU Shou-dong, XI Ya-jun

College of Agronomy, Northwest A&F University, Yangling, Shaanxi Province 712100, China

Received:2014-01-06 Revised:2014-04-17 Online:2014-12-15 Published:2014-12-01

摘要/Abstract

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P_SAG12-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P_SAG12-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P_SAG12-IPT基因在部分转基因植株中已经表达.

关键词: 柳枝稷, P_SAG12-IPT基因, 遗传转化, 农杆菌

Abstract:

In order to prolong the leaf functional period of bioenergy crop switchgrass, the leaf senescence gene P_SAG12-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 P_SAG12-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 P_SAG12-IPT gene was expressed in some of the transgenic plants.

Key words: Switchgrass, P_SAG12-IPT, Transformation, Agrobacterium

中图分类号:

柴乖强, 徐开杰, 王勇锋, 孙风丽, 刘曙东, 奚亚军. 农杆菌介导的P_SAG12-IPT基因转化柳枝稷研究[J]. 草地学报, 2014, 22(6): 1331-1336.

CHAI Guai-qiang, XU Kai-jie, WANG Yong-feng, SUN Feng-li, LIU Shou-dong, XI Ya-jun. Agrobacterium-mediated Transformation of P_SAG12-IPT Gene in Switchgrass[J]. Acta Agrestia Sinica, 2014, 22(6): 1331-1336.

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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

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农杆菌介导的PSAG12-IPT基因转化柳枝稷研究

Agrobacterium-mediated Transformation of PSAG12-IPT Gene in Switchgrass

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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