冰草组织培养再生体系建立及耐旱转基因研究

本研究以冰草属植物中的4个品种——蒙古冰草新品系(A.mongolicum Keng)、航道冰草(A.cristatum cv.Fairway)、诺丹冰草(A.desertorum cv.Nordan)和一个种间杂种冰草——蒙农杂种冰草(A.cristatum×A.desertorum cv.Hycrest-Mengnong)为材料,分别以幼穗为外植体建立了冰草组织培养再生体系。在此基础上,以调控脯氨酸生物合成最后一步的关键酶的突变体基因P5CS为目标基因,bar基因为筛选标记基因,用基因枪法轰击幼穗诱导的愈伤组织,获得转基因植株。利用PCR方法从拟南芥(Arabidopsis thaliana)基因组DNA中分离得到了CBF4及其启动子,并对CBF4启动子进行了序列分析和初步的功能分析。

Study on Culture and Regeneration of Wheatgrass and Its Genetic Transformation for Drought Tolerance

This paper tells how the wheatgrass tissue culture and regeneration system with the immature inflorescence as explant was established using four species, namely: A. mongolicum Keng, A.cristatum cv. Fairway, A.desertorum cv. Nordan, and A.cristatum×A.desertorum cv. Hycrest-Mengnong as materials. Based on the established regeneration system, the P5CS gene, which regulates the last step of key enzymes mutant gene of proline synthesis in a plant, was transformed into wheatgrass with phosphinothricin acetyltransferase (bar) conferring herbicide resistance as selecting gene. The transformation was conducted through microprojectile bombardment of callus derived from immature inflorescence.The CBF4 gene and its promoter had been isolated from Arabidopsis thaliana by PCR. The sequence analysis and function assay were conducted.