甜菜M14品系硫氧还蛋白过氧化物酶(BvM14-Tpx)基因在原核及真核细胞中的抗氧化及抗盐能力分析

Oxidation resistance and salt resistance analysis of Thioredoxin peroxidase(BvM14-Tpx) gene from Sugar Beet M14 line in transgenic prokaryotic and eukaryotic cells

甜菜M14品系是在栽培甜菜染色体基础上附加了野生白花甜菜第9号染色体的单体附加系,具有抗逆、无融合生殖等优异特性。在前期工作中,通过RACE技术获得了甜菜M14品系硫氧还蛋白过氧化物酶(Thioredoxin peroxidase,Tpx)编码基因BvM14-Tpx的cDNA全长,此基因参与各种过氧化物的清除反应从而提高植物的抗逆性。构建了BvM14-Tpx基因的大肠杆菌表达体系,对转BvM14-Tpx基因的大肠杆菌BL21(DE3)菌体在H2O2胁迫下的生长曲线进行了测定,结果显示转基因菌株比对照菌株提前1h进入生长期,说明BvM14-Tpx基因能够缓解H2O2对大肠杆菌生长的抑制;同时构建了BvM14-Tpx基因的酿酒酵母表达体系,对H2O2及NaCl胁迫下的转基因酿酒酵母的研究表明,转基因酵母的生长好于对照,说明BvM14-Tpx基因可以提高转基因酵母的抗氧化及耐盐能力。

Sugar beet monosomic addition line M14 was obtained from the intercross between a cultivated species Beta vulgaris L. and a wild species Beta corolliflora Zoss. It contains the Beta vulgaris L. genome with the addition of chromosome 9 of Beta corollif lora Zoss. The M14 line has exhibited the characteristics of stress tolerance and apomixes. Previously, a cDNA whole length of the BvM14-Tpx gene which coded for thioredoxin peroxidase was obtained by RACE. It was reported that the Tpx gene was involved in the removal of the various this study, the E. coli expression system peroxide reaction so as to improve the resistance of plants. In of BvM14-Tpx gene was built. The growth curve of E. coli BL21 （DE3） bacteria transferring BvM14-Tpx gene was measured under the stress of H2O2. The results showed that transgenic E. coli strains entered growing period 1 h earlier than the control strains, suggesting that BvM14-Tpx genes can ease the inhibition of H2O2 on the growth of E. coli. And the eukaryotic expression system of BvM14-Tpx gene was built. The results of transgenic Saccharomyces cerevisiae under H202 and NaC1 stress, showed that transgenic yeast grown better than the control, suggesting that BvM14-Tpx can improve the salt tolerance ability of transgenic yeast.