西伯利亚蓼非特异性脂质转移蛋白基因PsnsLTP的功能分析

Function of Non-specific Lipid Transfer Protein Gene PsnsLTP in Polygonum sibiricum Laxm

通过在农杆菌介导下,利用西伯利亚蓼非特异性脂质转移蛋白基因(植物表达载体:pROKII-PsnsLTP)对烟草进行遗传转化,经卡那霉素抗性分化筛选及PCR鉴定,获得6个转基因烟草株系。接种烟草炭疽病病菌和猝倒病病菌后观察转基因植株和野生型的表型差异,在Na Cl、Cd Cl2、7.5%PEG6000的胁迫下进行表型观察和生理指标的测定。结果显示:与野生型烟草相比,T1代转基因植株已具有较强的耐烟草炭疽病和猝倒病的能力;其超氧化物歧化酶(SOD)和过氧化物酶(POD)活性显著增强,丙二醛(MAD)含量显著降低,且幼苗具有较好的生长性状。由此证明,西伯利亚蓼非特异性脂质转移蛋白PsnsLTP增加了转基因烟草抵御生物胁迫和非生物胁迫的能力,这些工作为深入研究PsnsLTP基因的抗逆机制提供了参考。

We transformed the plant expression vector pROKII-PsnsLTP harboring non-specific lipid transfer protein gene from Polygonum sibiricum Laxm into tobacco by Agrobacterium-mediated genetic method. After transgenic tobacco was identified by kanamycin resistance screening and PCR, we observed phenotypic after induced Colletotrichum nicotianae and Pythium aphanidermatum, respectively, on the leaves of transgenic tobacco and wild type, and measured physiological indicators, the phenotypic in transgenic tobacco, and wild type in the treatments of NaC1, CdC12, 7.5% PEG6000 solutions. Compared with wild type tobacco, transgenic tobacco plants had a strong resistance to Colletotrichum nicotianae and Pythium aphanidermatum. SOD and POD activity was significantly enhanced, and MAD content was extremely reduced besides the transgenic tobacco seedlings had better growth traits. Therefore, PsnsLTP improved the ability of transgenic tobacco to resist biological and abiotic stress.