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Study of Heterologous Expression Hvsusiba2 Gene Increasing Rice (Oryza sotiva) Endosperm Starch Contents |
1,SHAN Zhen2,CHEN ZaiJie2,Wang Ying2 |
1. Fujian Provincial Key Laboratory of Genetic Engineering for Agriculture,Biotechnology Institute of Fujian Academy of Agricultural Sciences 2.
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Abstract Hvsusiba2 is a transcription factor acting on the upstream of starch synthesis pathway, and be recognized as a key regulator for barley (Hordeum vulgare) starch acumulation. In order to improve the rice (Oryza sotiva) grain starch content and understand how Hvsusiba2 works in rice, the Hvsusiba2 were introduced , which be constructed under down-stream of a specific expression promoter pSBEIIb from barley via agrobacterium-mediated transformation. Twenty-eight independent transformed lines were obtained. After self-crossing and screen, two homozygous lines with single-copy integrated were selected for investigation. By using qRT-PCR, genes involved in starch synthesis transcript level in 2 lines rice and non-transgenic control were detected. The results showed Hvsusiba2 was efficiently transcription in rice, along with 14 of genes involved in starch synthesis transcript were significantly altered. Among them 3 involved in amylopectin synthesis genes including starch branching enzyme gene (OssbeIIb), as well as isoamylase gene (Osiso1) and sucrose transporter gene (Ossut1) were elevated to above 2 time higher in the transgenic lines compared with the wild-type control. In additions 2 sucrose synthase genes (Ossus2, Ossus3), sucrose transporters gene (Ossut5) as well as UDP-glucose pyrophosphorylase gene (Osugp1) and starch branching enzyme gene (OssbeI) transcription were enhanced, leading to seeds starch content increased from 77.73% to 86.49% and 85.15% in 2 lines of Hvsusiba2 rice, respectively. Our data indicate that manipulation of starch in rice grain is possible using transcription factor. The results provide a theoretical basis for improving the content of rice endosperm starch by molecular method.
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Received: 05 July 2017
Published: 25 March 2018
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