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Development of high quality transgenic rice by gene targeting and overexpressing useful genes
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/297970
Rice is one of the most important major food crops which provide the major food for more than half of global population. To improve the grain quality as well as grain yield has been the essential breeding goal in rice. The composition of amylopectin is the determinant of rice eating quality under certain threshold of protein content and the ratio of amylose and amylopectin. In this study, RBE 1 driven by CaMV-35S promoter was constructed and transformed using Agrobacterium tumefaciens. We selected single copy with low amylose content among transgenic lines. The mRNA expression was investigated using RT-PCR, and enzyme activity was determined using activity staining method in mid-milky stage endosperm. Also, the overexpression vectors for RBE 1 and SSS 1 driven by seed specific globulin promoter were constructed, respectively. Moreover, the RNA interference vectors for soluble starch synthase 1 and granule bound starch synthase 1 derived by CaMV35S promoter were constructed, respectively and transformed using Agrobacterium tumefaciens. The transgene has been confirmed by amplification of HPT and target gene. The transgenic plants obtained will be used to investigate the gene function of related starch pathway in plant cells using Gopumbyeo as a wild type rice, based on the gain-of-function and the loss-of-function. The development of designed site-specific endonucleases boosted the establishment of gene targeting (GT) techniques in a row of different species. However, the methods described in plants require a highly efficient transformation and regeneration procedure and, therefore, can be applied to very few species. Here, we describe a highly efficient GT system that is suitable for all transformable plants regardless of transformation efficiency. Efficient in planta GT was achieved in rice by expression of a site specific endonuclease (SSS1::ZFN) that not only cuts within the target but also the chromosomal transgenic donor, leading to an excised targeting vector.
