摘要
以农杆菌介导的黄瓜遗传转化体系为基础,以抗冷基因BnCS为外源基因对黄瓜进行转化试验,对转化系统中影响植株再生的主要因素进行研究,结果表明,子叶节预培养1d,再生效果最好;用划伤的方法处理微伤口较好,切割边缘制造微伤口效果较差;侵染时间以20min为最佳,再生芽率达到52.6%;脱菌抗生素cef对芽分化的影响较大,降低再生率近20%。对本试验的再生植株进行PCR检测,结果显示,PCR阳性率为38.4%。
Some main factors affecting the efficiency of regeneration of Agrobacterium-mediated transformation of cucumbers in which the gene BnCS was transferred are studied in the research. The results indicated that: When pre-cultured the cotyledon nodes for ld, the best efficiency of regeneration was obtained. Furthermore it's better to scratch the cotyledon nodes than not to do that, while the worst result was gained when only scratch the edge of the cotyledon. The regeneration rate was 52.6% when infected for 20min. However the antibiotic cef decreased the regeneration rate for about 20%. The regenerated plants were tested by PCR and 38.4% of them were positive.
基金
黑龙江省自然科学基金资助项目(C200502)