数字农科院2.0

Editing Of Rice Isoamylase Gene I.sa1 Provides Insights Into I ts Function In Starch Formation

文献类型： 外文期刊

作者： Luo Ju;Jiao Guiai;Sheng Zhonghua;Cai Yicong;Wang Jianlong;Tang Shaoqing;Feng Baobing;Chao Shufen;Hu Peisong;Tang Shaoqing;Wei Xiangjin

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关键词： ISA1; rice; starch biosynthesis; starch granule; physicochemical property; CRISPR; Cas9

期刊名称： RICE SCIENCE

ISSN： 1672-6308

年卷期： 2019 年 26 卷 2 期

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摘要： Isoamylase 1 (ISA1) is an isoamylase-type debranching enzyme which plays a predominant role in amylopectin synthesis. In this study, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonuclease 9 (CRISPR/Cas9) system was used to edit ISA1 gene in rice via Agrobacterium-mediated transformation. We identified 36 genetic edited lines from 55 T-0 transgenic events, and classified the mutation forms into 7 types. Of those, two homozygous mutants, cr-isa1-1 (type 1, with an adenine insertion) and cr-isa1-2 (type 3, with a cytosine deletion) were selected for further analysis. Seed sizes of both cr-isa1-1 and cr-isa1-2 were affected, and the two mutants also displayed a shrunken endosperm with significantly lower grain weight. Electron microscopy analysis showed that abnormal starch granules and amyloplasts were found in cr-isa1-1 and cr-isa1-2 endosperm cells. The contents of total starch, amylose and amylopectin in the endosperm of the cr-isa1 mutants were significantly reduced, whereas sugar content and starch gel consistency were observably increased compared to the wild-type. The gelatinization temperature and starch chain length distributions of the cr-isa1 mutants were also altered. Moreover, transcript levels of most starch synthesis-related genes were significantly lower in cr-isa1 mutants. In conclusion, the results indicated that gene edition of ISA1 affected starch synthesis and endosperm development, and brought potential implications for rice quality breeding.

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