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为改良草坪型高羊茅(Festuca arundinacea Schreb.)的耐逆性,以成熟种子来源的胚性愈伤组织为受体材料,通过农杆菌介导法将拟南芥(Arabidopsis thaliana)耐逆相关CBF1基因导入4个供试品种的基因组,经GUS染色、PCR检测和Southern杂交分析验证,获得了112株转基因植株,转化频率为0.92%~2.87%,不同品种间存在差异。试验表明,在高盐与高渗胁迫下,转基因植株具有显著生长优势,存活率极显著高于非转化对照植株,经低温、高温、干旱和高盐等逆境胁迫处理后的叶片相对电导率平均较对照植株低25%~30%,证明转基因植株的耐逆性有所增强。考察发现,非胁迫条件下CBF1基因的组成型超表达使转基因植株的生长受到抑制。
In order to improve the tolerance of the lawn-type tall fescue (Festuca arundinacea Schreb.), The embryogenic callus derived from mature seeds was used as the receptor material, and the resistance-resistant relationship of Arabidopsis thaliana was mediated by Agrobacterium tumefaciens The CBF1 gene was introduced into the genomes of 4 tested cultivars. After GUS staining, PCR and Southern hybridization analysis, 112 transgenic plants were obtained. The frequency of transformation was 0.92% -2.87%. There were differences among different cultivars. The results showed that under high salt and high osmotic stress, the transgenic plants had significant growth advantages, and the survival rate was significantly higher than that of the non-transformed plants. The average relative conductivity of the leaves under the stress of low temperature, high temperature, drought and high salt 25% to 30% lower than that of the control plants, which proves that the tolerance of the transgenic plants is enhanced. It was found that the constitutive overexpression of CBF1 gene inhibited the growth of transgenic plants under non-stress conditions.