转入盐地碱蓬谷胱甘肽转移酶和过氧化氢酶基因增强水稻幼苗对低温胁迫的抗性

Transferring the Suaeda salsa Glutathione S-Transferase and Catalase Genes Enhances Low Temperature Stress Resistance in Transgenic Rice Seedlings

以水稻(OryzasativaL.)品种中花11号成熟种子为材料,利用农杆菌介导法将盐地碱蓬的GST(谷胱甘肽转移酶)单基因和GST+CAT1(catalase1)双基因转入低温敏感水稻品种中花11号,并对T4代转基因水稻幼苗的抗低温特性进行了分析。结果显示,低温处理后,转基因植株的GST和CAT活性都比未转入这两种基因的对照高;且PSII最大光化学效率也高于非转基因对照;而H2O2和MDA(malondialdehyde)的含量及细胞膜透性则低于对照。说明转基因水稻幼苗GST和GST+CAT1的表达提高了对低温胁迫的抗性。

The GST （glutathione S-transferase） and GST＋CAT1 （catalase 1） of Suaeda salsa were introduced into a low temperature-sensitive rice cultivar （Oryza sativa cv. Zhonghua No. 11） by Agrobacterium tumefaciens-mediated transformation under the control of cauliflower mosaic virus （CaMV） 35S promoter, and the transformed calli and plantlets were screened on Murashige and Skoog （1962） medium supplemented with hygromycin 25 μg/mL and cefotaxime 300 μg/mL. The putative primary transformants （T0 generation） were acclimatized at 26℃/22℃ in a greenhouse for 7 d, and then transplanted to the field, where they grew up to maturity under outdoor conditions. 25 and 14 independent transgenic lines of T1 generation carrying the GST and GST＋ CAT1 genes, respectively, were identified by PCR amplification. Transgene expression was monitored by RNA-blot hybridization using total RNA samples from leaf tissues （Figs. 1, 2）. To investigate whether express- ing the Suaeda salsa GST and GST＋CAT1 in transgenic rice increased low temperature stress tolerance, the T4 14-day-old transgenic and non-transgenic rice seedlings were transferred to a low temperature （day 7℃/night 4℃） growth chamber for 3-6 d. The experimental data showed that expressing the Suaeda salsa GST and GST＋CAT1 enhanced low temperature stress resistance in transgenic rice seedlings. When treated with low temperature, both GST and CAT activity increased in the transformants with the time of temperature treatment （Figs.8, 9A,B）. These transgenic rice plant seedlings exhibited a higher level of photosynthetic capacity than those of the non-transgenic control seedlings under low temperature treatment （Fig.3）. Whereas, there were lower H2O2 （Fig.4） and MDA （malondialdehyde） content （Fig.6）, and relative electrolyte leakage through the plasma membrane was also lower in transgenic rice seedlings than in the parent line under low temperature condition （Fig.5）. The results also indicated that GST＋CAT1 co-expression conferred greater level of low temperature stress tolerance to the transformed rice plants compared to the single GST transformed plants.