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构建了同时含有胞质谷氨酰胺合成酶(GS1)cDNA和叶绿体谷氨酰胺合成酶(GS2)cDNA的植物表达载体p2GS,通过农杆菌介导法用它们转化了水稻品种“中花10号”的成熟胚愈伤组织,经潮霉素(Hyg)筛选培养及分化再生,获得了抗Hyg的转基因水稻植株。PCR和基因组Southern杂交分析结果证明,GS1和GS2基因均已经整合到转基因水稻的基因组内。Northern杂交实验结果证实,GS1和GS2基因在转基因水稻的转录水平上得到了有效表达。在以0.7mmol/L的(NH4)2SO4取代了其中氮成分的MS培养基上测试植株生长量,结果表明转基因植株鲜重增长量显著高于对照,证明高效表达GS增强了转基因水稻对土壤氮素缺乏的耐性。
The plant expression vector p2GS containing both the cytoplasmic glutamine synthetase (GS1) cDNA and the chloroplast glutamine synthetase (GS2) cDNA was constructed and transformed into rice “Zhonghua 10” by Agrobacterium tumefaciens- Of mature embryonic callus, Hyg screening and differentiation and regeneration, obtained anti-Hyg transgenic rice plants. PCR and genomic Southern hybridization analysis results show that GS1 and GS2 genes have been integrated into the genome of transgenic rice. Northern blot results confirmed that the GS1 and GS2 genes were efficiently expressed at the transcriptional level of transgenic rice. The plant growth was tested on MS medium with 0.7 mmol / L (NH4) 2SO4 instead of nitrogen. The results showed that the fresh weight increase of transgenic plants was significantly higher than that of the control, indicating that high efficiency of GS enhanced the effect of transgenic rice on soil nitrogen Lack of patience.