【目的】建立提取鱼腥草叶片高质量RNA的有效方法。【方法】采用RNAiso for Polysaccharide-rich Plant Tissue法、十六烷基三甲基溴化铵(CTAB)法、十二烷基硫酸钠(SDS)Ⅰ法和SDSⅡ法提取鱼腥草叶片总RNA。在匀浆上清液抽提时分别选用6种试剂组合,通过现象观察,检测紫外吸光度值和琼脂糖凝胶电泳图,考察RNA的纯度、产率和完整性,比较各方法提取效果的差异。【结果】RNAiso for Polysaccharide-rich Plant Tissue法、CTAB法和SDSⅡ法使用苯酚/氯仿/异戊醇或NaCl加苯酚/氯仿/异戊醇抽提后,所得RNA纯度和完整性均好,最大得率为172.6μg/g。【结论】确定了鱼腥草总RNA提取的有效方法,为鱼腥草转录组研究打下基础。
[Objective] This study was to identify the expression of exogenous antimicrobial peptide in transgenic Houttuynia cordata Thunb. plants,and analyze their resistance to stem rot disease. [Methods] SDS-PAGE and Western blot analysis were employed to detect expression of exogenous antimicrobial peptide in transgenic H. cordata plants. Both wild type and transgenic H. cordata plants were inoculated with different concentrations of Rhizoctonia solani spores for detecting their resistance. [Results] The exogenous antimicrobial peptide was detected at translation level. The optimal parameters for detecting the resistance of transgenic H. cordata plants to R. solani was inoculation of spores at a concentration of 3×105 ind./ml and cultured for three days. The results showed that resistance of transgenic H. cordata plants to R. solani was enhanced in comparison with CKs. [Conclusion] Expression of exogenous antimicrobial peptide can enhance the resistance of transgenic H. cordata plants to stem rot disease.
