论文部分内容阅读
从耐盐性强的红树植物白骨壤(Avicennia marina)中分离出的耐盐基因CSRG1,构建了pGAM189/CSRG1转基因载体,以烟草嫩叶为外植体,通过农杆菌介导的叶盘转化法将CSRG1基因及GUS基因,Kmr基因和Hygr基因转入烟草基因组中.转化50个外植体,经过50 mg/L潮霉素和150mg/L卡那霉素抗性筛选共获得了13个稳定抗性株系.通过PCR扩增检测、southern blot分析和GUS基因活性检测,结果表明,最终获得的11个转基因品系(基因转化频率为22％)中,CSRG1基因整合到烟草的染色体DNA上.Northern blot分析结果表明,CSRG1基因在转基因植株中获得表达.耐盐性测定和光合速率测定结果表明,转基因植株在盐分提高到2％NaCl和全海水(盐度为24)配置的MS培养基中,成活率保持在80％-90％,株高增长20％-40％,CSRG1基因的表达产物及形成的生理代谢途径确实可使烟草获得较高的耐盐性,同时这种耐盐性不仅针对钠离子胁迫,而且对于各种离子的综合盐胁迫都具有耐受性.
The salt-tolerant gene CSRG1 was isolated from the salt-tolerant mangrove Avicennia marina, and the pGAM189 / CSRG1 transgenic vector was constructed. The young tobacco leaves were used as explants and Agrobacterium -mediated leaf disc transformation The CSRG1 gene, GUS gene, Kmr gene and Hygr gene were transferred into the tobacco genome.The transformation of 50 explants was screened by 50 mg / L hygromycin and 150 mg / L kanamycin resistance for 13 Stable resistant strains.The results of PCR amplification, southern blot analysis and GUS gene activity showed that the CSRG1 gene was integrated into the chromosomal DNA of tobacco in 11 transgenic lines (gene transformation frequency of 22%) Results of Northern blot analysis showed that CSRG1 gene was expressed in transgenic plants.The results of salt tolerance and photosynthetic rate indicated that the transgenic plants were cultured in MS medium with 2% NaCl and 24% , Survival rate remained at 80% -90%, plant height increased by 20% -40%, CSRG1 gene expression products and the formation of the physiological metabolic pathway can really get higher salt tolerance of tobacco, while the salt tolerance Not only for sodium ion stress, but also Integrated salt stress in various ions are resistant.