论文部分内容阅读
【目的】为了提高珠美海棠的耐盐性，建立更高效的珠美海棠遗传转化体系。【方法】在转化液中利用超声波直接转化在无菌条件下横切主脉的珠美海棠试管苗幼叶，通过愈伤组织培养诱导植株再生、抗性筛选，GUS 染色鉴定，以及 RT-PCR 分析。【结果】试验结果表明，较利于珠美海棠愈伤组织诱导分化及抗性筛选的培养基为：MS+6-BA2.0 mg/L + NAA 0.1 mg/L+Kana50 mg/L。超声波处理的最优工作条件为：处理6 s，间歇10 s，工作重复20次，功率80 w，转化效率为31.3%。GUS 染色与 RT-PCR 鉴定结果一致率为100%，RT-PCR 结果分析显示，Mz2NHX1基因在转化苗中的表达量约为对照的3倍。荧光定量 PCR 结果显示转化苗 Mz2NHX1基因的表达量为对照组的6.21倍，盐处理试验结果显示转化苗耐盐性得到了显著地提高。【结论】建立了超声波介导的珠美海棠高效遗传转化体系，获得珠美海棠耐盐新材料，为该耐盐基因的研究和盐碱地的改良奠定了基础。
【Objective】 In order to improve the salt tolerance of Begonia tuberosus, a more efficient genetic transformation system was established. 【Method】 The young leaves of Begonia Begonia were transected under sterile conditions by ultrasonic in the transformation solution. Plant regeneration was induced by callus induction, resistance screening, GUS staining and RT-PCR analysis . 【Result】 The results showed that the medium which was more suitable for callus differentiation and resistance screening of M. crabunia was: MS + 6-BA 2.0 mg / L + NAA 0.1 mg / L + Kana 50 mg / L. The optimal working conditions of ultrasonic treatment were as follows: processing 6 s, intermittent 10 s, repetitive work 20 times, power 80 w, conversion efficiency 31.3%. The coincidence rate of GUS staining and RT-PCR results was 100%. The results of RT-PCR showed that the expression level of Mz2NHX1 in transformed seedlings was about 3 times of that in control. Fluorescent quantitative PCR results showed that the expression level of Mz2NHX1 gene in transformed plants was 6.21 times that of the control group, and the result of salt treatment showed that salt tolerance of transformed plants was significantly increased. 【Conclusion】 The ultrasonic-mediated genetic transformation system of Begonia tuberosus was established, and the salt-tolerant new material of Begonia tuberosus was established, which laid the foundation for the study of salt tolerance gene and the improvement of saline-alkali soil.