转TSRF1基因尾叶桉的培育及其广谱抗病性

Transformation of TSRF1 into Eucalyptus urophylla and the Broad-Spectrum Disease Resistance of the Transgenic Plant to Diseases

以尾叶桉U6无性系无菌种子苗下胚轴为外植体,通过对不同生长调节剂的优化组合,建立尾叶桉高效再生体系。应用正交试验设计,优化尾叶桉遗传转化体系。经PCR和Southern杂交检测表明,TSRF1基因已整合到尾叶桉基因组中;转基因植株灌根接种青枯菌后,转基因植株表现出发病延迟,抗性提高,抗性相关酶活性显著增强。转化植株离体叶片对疫霉菌抗性明显增强。荧光定量PCR的结果表明转基因植株接种致病菌后TSRF1基因表达量得到提高。研究结果表明,转TSRF1基因尾叶桉表现出一定的广谱抗病能力。

In this study, the hypocotyls germinated from the seeds of Eucalyptus urophylla U6 were used as explants, and the different plant growth substance combinations for the regeneration and conditions for genetic transformation of E. urophylla were optimized by an orthogonal test. The TSRF1 gene was confirmed to integrate into the E. urophylla genome by PCR and Southern blotting. The transgenic E. urophylla markedly enhanced the disease resistance to Ralstonia solanacearum. The transgenic plants had stronger resistance to the leaf disease infested with Phytophthora. After inoculation with R. solanacearum, the activity of PPO, PAL and SOD significantly increased. The expression of TSRF1 gene was increased after inoculation with diseases, detected by the method of Real-time PCR. These results suggested that expression of the TSRF1 gene could improve broad-spectrum disease resistance in the transgenic E. urophylla.