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Agrobacterium-mediated transformation of sorghum and analysis of putative transposable element-induced mutants in sorghum.

机译:农杆菌介导的高粱转化和高粱中可能的转座因子诱导的突变体的分析。

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Plant biotechnology has become a powerful tool to complement the traditional methods of plant improvement. Several methodologies have been developed to identify and clone agronomically important genes and to transfer genes from any living organism to plants. This work encompasses the study of two methodologies related to the identification and transferring of genes in sorghum. The first part reports the development of a protocol for sorghum transformation via Agrobacterium tumefaciens. It is demonstrated that Agrobacterium-mediated transformation is a feasible technique for the genetic transformation of sorghum. Sorghum transgenic plants were produced via Agrobacterium tumefaciens, and the transformation evidenced by Southern blot analysis of T0 and T1 plants, detection of GUS activity, and production of T1 plants resistant to hygromycin. Immature embryos of sorghum were very sensitive to Agrobacterium, and embryo death after co-cultivation was considered the limiting step to increase the transformation efficiency. Key factors were the co-cultivation medium, the use of a genotype and an explant with good tissue culture response, and the addition of Pluronic F-68 to the inoculation medium. Sorghum transformation via Agrobacterium is still not a routine technique, but it seems to have good potential once the protocol is further refined and improved.; The second part of the thesis describes the analysis of several putative transposable element-induced mutants in sorghum identified in a Candystripe population, and the identification of endogenous transposable elements in sorghum derived from the cs1 family. Lastly, a likely involvement of a transposable element in the susceptibility of Colby to Periconia circinata is discussed.
机译:植物生物技术已成为补充传统植物改良方法的有力工具。已经开发出几种方法来鉴定和克隆农学上重要的基因并将基因从任何活生物体转移到植物。这项工作包括对与高粱基因鉴定和转移有关的两种方法学的研究。第一部分报告了通过 Agrobacterium tumefaciens 转化高粱的方法的发展。结果表明, Agrobacterium 介导的转化是高粱遗传转化的可行技术。通过根癌土壤杆菌(Agrobacterium tumefaciens)产生高粱转基因植物,并通过T 0 和T 1 植物的Southern印迹分析,检测GUS活性证明了转化,并产生对潮霉素具有抗性的T 1 植物。高粱的未成熟胚对农杆菌非常敏感,共培养后的胚胎死亡被认为是提高转化效率的限制步骤。关键因素是共培养培养基,具有良好组织培养响应的基因型和外植体的使用以及在接种培养基中添加Pluronic F-68。通过农杆菌进行的高粱转化仍然不是常规技术,但是一旦进一步完善和改进该协议,它似乎具有良好的潜力。论文的第二部分描述了在Candystripe群体中鉴定出的高粱中几种假定的可转座因子诱导的突变体的分析,以及鉴定了来自 cs1 家族的高粱中的内源转座因子。最后,讨论了转座因子可能参与Colby对 Periconia circinata 的易感性。

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