Plant Regeneration and Its Functional Analysis within Transgenic Rice of ASG-1, an Apomixis-specific Gene Isolated from Apomictic Guinea Grass
Journal Title: Biotechnology Journal International - Year 2016, Vol 16, Issue 3
Abstract
Aims: To analyze the function of the ASG-1, an apomixis-specific gene has been isolated from apomictic guinea grass, the combination of Agrobacterium-mediated transformation system and pSMA35H2-NG binary vector were used for transformation of ASG-1 to ‘Nipponbare’ of rice, and its functional analysis of transgenic rice was carried out. Study Design: 1) plant regeneration of transgenic rice of ASG-1 was achieved, and 2) it was followed by experiments with the obtained non-graining ear in the whole plants (NGEP) using comparative observation in both pollens of guinea grass and the T0 NGEP with Nomarski differential interference-contrast microscopy (DIC). Place and Duration of Study: Faculty of Environmental and Horticultural Science, Minami Kyushu University, between September 2012 and October 2015. Methodology: As a preliminary transformation, a β-glucuronidase (GUS) was used to infect the callus. Plant regeneration of the transgenic rice of ASG-1 were achieved. DNAs of the transgenic rice were used for detection of ASG-1 by PCR. For analysis of NGEP, the anthers and ovaries of the transgenic rice and guinea grass were used for comparative observation with DIC. Results: 1) GUS was successfully expressed in calli of both rice and guinea grass. 2) The transgenic rice plants of ASG-1 were then obtained from the culture of seed-derived calli. 3) The natures of T0 callus and T1 plants were confirmed. 4) A subset (~15%) of T0 plants showed morphological sterility with NGEP. 5) The ovaries of the NGEP showed identical characteristics to “Nipponbare” and other normal T1 plants; however, the pollen showed normal rates of <38%. 6) The pollen of guinea grass at anthesis showed rates of normal pollen as low as that of the NGEP rice. Conclusion: ASG-1 may play a role in regulating pollen sterility. Comparing these findings with our previous reports, we identified co-localization for ASG-1 in pollen of both rice and guinea grass, suggesting that the trait of ASG-1 influencing pollen sterility might be inherited from guinea grass to rice through the transformation process.
Authors and Affiliations
Lanzhuang Chen, Liming Guan, Daiki Toyomoto, Toru Sugita, Takuro Hamaguchi, Reiji Okabe
Keywords
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- EP ID EP237924
- DOI 10.9734/BJI/2016/29146
- Views 119
- Downloads 0
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