Transformed Rice with Salt Tolerance-related Genes of Bruguiera sexangula by Agrobacterium Meditation

Abstract

运用农杆菌介导法将红树林耐盐相关基因mangrin转入粳稻品种‘日本晴’中,通过GUS基因检测愈伤组织转化率,确定农杆菌菌液浓度OD600为0.5,浸染时间30min,共培养时间3d为最佳转化体系;经潮霉菌筛选,获得抗性再生植株。通过PCR扩增检测、Southern blot分析和GUS基因活性检测,结果表明,mangrin基因整合到再生水稻的染色体DNA上。耐盐性测定结果表明,转基因植株在200mmol/L NaCl胁迫下,成活率保持在83.3%,株高增长20%~40%,mangrin基因能提高转基因水稻对盐胁迫的抗性。Salt tolerance-related genes in Bruguiera sexangula were transformed into Oryza sativa subsp.japonica cv.Nipponbare,a keng-rice variety.The optimal system of the transformation was found by detecting the callus-transformation rate with GUS gene to be the one with the agro-bacterium concentration set at OD_ 600 0.5,the infection time set at 30 minutes and the culture time set at 3 days and then the regeneration plantlets were obtained by Hyg screening.mangrin gene was integrated in the DNA of the regeneration plantlets by PCR amplification,Southern blotting and GUS gene activity detection.The testing of the salt tolerance indicated that under the stress resulting from 200 mmol/L NaCl the transgenic plants maintained a survival rate of 83.3% and increased their heights by 20%~40% and thus mangrin gene could raise the resistance of the transformed rice to salt stress.中国科学院知识创新工程重要方向项目(KZCX3-SW-444);; 教育部重点项目(01102

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