摘要
本研究采用RT-PCR方法克隆得到水稻细胞质抗坏血酸过氧化物酶(OsAPX1)基因全长cDNA(基因登录号:D45423);将该基因构建二元植物表达载体pBI121-OsAPX1;用农杆菌EHA105侵染介导烟草叶盘转化法转基因,获得转基因植株,PCR鉴定遗传转化的烟草植株成功地整合了水稻OsAPX1基因;Northern blot鉴定结果表明转基因T1代植株Ta在mRNA转录水平上表达;对Ta株系进行Kana抗性鉴定,表明转基因T2代株系有抗性;对T2代株系做NaCl、NaHCO3、Na2CO3抗盐性鉴定,结果表明与对照相比表现出转基因植株抗(耐)性提高;取T2代植株的叶片在不同浓度H2O2处理下,抗H2O2毒害的能力显著强于对照,转基因植物的抗性有所提高,有望在抗盐碱性育种上应用。
In order to explore the role of ascorbate peroxidase in plant during salt stress, the full-length rice cytoplasm ascorbate peroxidase gene （OsAPX1） was cloned by RT-PCR amplification and constructed into a binary vector pBI121-OsAPX1. The OsAPX1 gene was introduced into tobacco by Agrobaterium mediated transformation. PCR identification of transformed tobacco plant showed that the OsAPX1 gene was integrated into the tobacco genome. Northern blot analysis indicated that T1 transformed plants Ta, expressed in mRNA level. Ta transformants were identified by Kan-resistance screening, showed the T2 transformed plants were tolerant to Kanamicin. Alkalinity-salinity tolerance identification of T2 transformants showed higher resistance to NaC1, NaHCO3 and Na2CO3, respectively, in comparison with controls. Leaves of T2 transformed plant were treated with a variety concentration of H2O2 and revealed higher resistance at H2O2, compared to controls. These indicated that the tolerance of transformed plant was improved and might be contributed in plant alkalinity-salinity resistance breeding.
出处
《分子植物育种》
CAS
CSCD
2007年第1期1-7,共7页
Molecular Plant Breeding
基金
国家自然科学基项目(30140003)
黑龙江省科技计划项目(GB04B606-02-02)资助。
关键词
抗坏血酸过氧化物酶
转基因
活性氧
烟草
Ascorbate peroxidase, Gene transformation, Reactive oxygen species, Tobacco
