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      Mutation of RGG2, which encodes a type B heterotrimeric G protein γ subunit, increases grain size and yield production in rice

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          Summary

          Heterotrimeric G proteins, which consist of G α, G β and G γ subunits, function as molecular switches that regulate a wide range of developmental processes in plants. In this study, we characterised the function of rice RGG2 , which encodes a type B G γ subunit, in regulating grain size and yield production. The expression levels of RGG2 were significantly higher than those of other rice G γ‐encoding genes in all tissues tested, suggesting that RGG2 plays essential roles in rice growth and development. By regulating cell expansion, overexpression of RGG2 in Nipponbare ( NIP) led to reduced plant height and decreased grain size. By contrast, two mutants generated by the clustered, regularly interspaced, short palindromic repeat ( CRISPR)/CRISPR‐associated protein 9 (Cas9) system in the Zhenshan 97 ( ZS97) background, zrgg2‐1 and zrgg2‐2, exhibited enhanced growth, including elongated internodes, increased 1000‐grain weight and plant biomass and enhanced grain yield per plant (+11.8% and 16.0%, respectively). These results demonstrate that RGG2 acts as a negative regulator of plant growth and organ size in rice. By measuring the length of the second leaf sheath after gibberellin ( GA 3) treatment and the GA‐induced α‐amylase activity of seeds, we found that RGG2 is also involved in GA signalling. In summary, we propose that RGG2 may regulate grain and organ size via the GA pathway and that manipulation of RGG2 may provide a novel strategy for rice grain yield enhancement.

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          Most cited references58

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          Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.

          A large number of morphologically normal, fertile, transgenic rice plants were obtained by co-cultivation of rice tissues with Agrobacterium tumefaciens. The efficiency of transformation was similar to that obtained by the methods used routinely for transformation of dicotyledons with the bacterium. Stable integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis of transformants in the R0, R1 and R2 generations. Sequence analysis revealed that the boundaries of the T-DNA in transgenic rice plants were essentially identical to those in transgenic dicotyledons. Calli induced from scutella were very good starting materials. A strain of A. tumefaciens that carried a so-called 'super-binary' vector gave especially high frequencies of transformation of various cultivars of japonica rice that included Koshihikari, which normally shows poor responses in tissue culture.
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            Assaying chimeric genes in plants: The GUS gene fusion system

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              GS3, a major QTL for grain length and weight and minor QTL for grain width and thickness in rice, encodes a putative transmembrane protein.

              The GS3 locus located in the pericentromeric region of rice chromosome 3 has been frequently identified as a major QTL for both grain weight (a yield trait) and grain length (a quality trait) in the literature. Near isogenic lines of GS3 were developed by successive crossing and backcrossing Minghui 63 (large grain) with Chuan 7 (small grain), using Minghui 63 as the recurrent parent. Analysis of a random subpopulation of 201 individuals from the BC3F2 progeny confirmed that the GS3 locus explained 80-90% of the variation for grain weight and length in this population. In addition, this locus was resolved as a minor QTL for grain width and thickness. Using 1,384 individuals with recessive phenotype (large grain) from a total of 5,740 BC3F2 plants and 11 molecular markers based on sequence information, GS3 was mapped to a DNA fragment approximately 7.9 kb in length. A full-length cDNA corresponding to the target region was identified, which provided complete sequence information for the GS3 candidate. This gene consists of five exons and encodes 232 amino acids with a putative PEBP-like domain, a transmembrane region, a putative TNFR/NGFR family cysteine-rich domain and a VWFC module. Comparative sequencing analysis identified a nonsense mutation, shared among all the large-grain varieties tested in comparison with the small grain varieties, in the second exon of the putative GS3 gene. This mutation causes a 178-aa truncation in the C-terminus of the predicted protein, suggesting that GS3 may function as a negative regulator for grain size. Cloning of such a gene provided the opportunity for fully characterizing the regulatory mechanism and related processes during grain development.
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                Author and article information

                Contributors
                ricegb@yzu.edu.cn
                zhouyong@yzu.edu.cn
                Journal
                Plant Biotechnol J
                Plant Biotechnol. J
                10.1111/(ISSN)1467-7652
                PBI
                Plant Biotechnology Journal
                John Wiley and Sons Inc. (Hoboken )
                1467-7644
                1467-7652
                13 December 2018
                March 2019
                : 17
                : 3 ( doiID: 10.1111/pbi.2019.17.issue-3 )
                : 650-664
                Affiliations
                [ 1 ] Jiangsu Key Laboratory of Crop Genetics and Physiology/Co‐Innovation Center for Modern Production Technology of Grain Crops Key Laboratory of Plant Functional Genomics of the Ministry of Education Yangzhou University Yangzhou China
                [ 2 ] Institute of Food Crops Jiangsu Academy of Agricultural Sciences Nanjing China
                [ 3 ] Shanghai Academy of Agricultural Sciences Shanghai China
                Author notes
                [*] [* ] Correspondence (Tel 86‐514‐87937619; fax 86‐514‐87972138; email zhouyong@ 123456yzu.edu.cn (YZ) and Tel 86‐514‐87972138; fax 86‐514‐87972138; email ricegb@ 123456yzu.edu.cn (GL))
                [†]

                These authors contributed equally to this work.

                Author information
                http://orcid.org/0000-0002-0434-1617
                Article
                PBI13005
                10.1111/pbi.13005
                6381795
                30160362
                a12c6a35-64d2-4791-ace1-4a9872050a42
                © 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 21 March 2018
                : 20 August 2018
                : 23 August 2018
                Page count
                Figures: 11, Tables: 1, Pages: 15, Words: 10229
                Funding
                Funded by: Natural Science Foundation of Jiangsu Province
                Award ID: BK20161335
                Funded by: National Key Research and Development Programme
                Award ID: 2016YFD0100400
                Funded by: University Science Research Project of Jiangsu Province
                Award ID: 15KJA210003
                Funded by: Priority Academic Program Development of Jiangsu Higher Education Institutions
                Funded by: China Postdoctoral Science Foundation
                Award ID: 2016M601899
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                pbi13005
                March 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.5.9 mode:remove_FC converted:20.02.2019

                Biotechnology
                heterotrimeric g protein,rgg2,rice,grain size,yield production
                Biotechnology
                heterotrimeric g protein, rgg2, rice, grain size, yield production

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