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Characterization and cloning of a brittle culm mutant (bc88) in rice (Oryza sativa L.)

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Chinese Science Bulletin 2013年第24期58卷 3000-3006,2914页

作者： rao yuchun YANG YaoLong XIN DeDong LI XiaoJing ZHAI KaiEn MA BoJun PAN JianWei QIAN Qian ZENG DaLi College of Chemistry and Life Sciences Zhejiang Normal University State Key Laboratory of Rice Biology China National Rice Research Institute

This study characterizes a brittle culm (bc88) mutant of rice (Oryza sativa L.) obtained by ethylene methylsulfonate (EMS)-induced mutagenesis of Wuyunjing 7. The bc88 mutant exhibits a diversity of pleiotropic phenotypes, including brittle culm at the whole-plant growth stages, withered leaf tips at the seedling stage, and 18-d delay in heading date at the mature stage. Genetic analysis indicates that the bc88 mutant is controlled by a single recessive nuclear gene. The mutated bc88 gene isolated by map-based cloning contains only one point mutation in the 5th exon relative to its wild-type BC88 (LOC_Os09g25490 and Os09g0422500), leading to an amino acid change from P to L in bc88 plants. Alignment of the putative protein sequence with its homologs indicates that the mutation is located in the conserved region of the sequence. Detection of the transcription level of BC88 in rice plants shows that the expression level of BC88 is higher in spikes and culms than in leaves, roots, and leaf sheaths. These contribute to understanding of the molecular mechanism of cellulose synthesis. The target gene BC88 can be a useful tool in molecular marker-assisted selection for rice culm trait breeding.

关键词： rice (Oryza sativa L.) brittle culm Wuyunjing 7 clone mutation

A Method for Effectively Overcoming Tight Functional Linkage Between Genes in Rice by CRISPR/Cas9 System

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Rice science 2020年第3期27卷 180-183页

作者： LI Sanfeng SHEN Lan HU Ping WU Xianmei YUAN Qiaoling rao yuchun QIAN Qian WANG Kejian ZHU Xudong SHANG Lianguang WANG Yuexing State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural SciencesShenzhen 518120China College of Chemistry and Life Sciences Zhejiang Normal UniversityJinhua 321004China

Anthocyanins are widely distributed in one or more parts of rice(Oryza sativa L.)plants,including seed coat,stigma,apiculus,leaf sheath and leaf blade,and are the main pigments used in rice to achieve different colors(Hou et al,2009;Aizza and Dornelas,2011).In rice,tissue-specific color traits(especially the color of apiculus,namely the lemma and palea of the spikelet)are not only important for rice variety identification but also important for linkage analysis and rice domestication research(Saitoh et al,2004;Fan et al,2007;Lin et al,2019).The apiculus color is controlled by the complementary functions of three pairs of dominant genes,C,A and *** C(chromogen)is a pigment gene,which is the basic gene for producing *** A(activator)activates gene C,converting the chromogen into anthocyanins,and gene P(purple)controls the distribution of anthocyanins in various organs(Reddy,1996;Sakamoto et al,2001).

关键词： CRISPR/Cas9 linkage blade

MULTI-TILLERING DWARF1, a new allele of BRITTLE CULM 12, affects plant height and tiller in rice

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Science Bulletin 2016年第23期61卷 1810-1817页

作者： Haiping Yu Deyong Ren Yangzhou Zhu Jiangmin Xu Yuexing Wang Ruifang Liu Yunxia Fang Zhenyuan Shi Jiangjie Pan Mei Lu Bojun Ma Jiang Hu yuchun rao College of Chemistry and Life Sciences Zhejiang Normal University State Key Laboratory for Rice Biology China National Rice Research Institute Key Laboratory of Northeast Rice Biology and Breeding Ministry of Agriculture and Key Laboratory of Northern Japonica Rice Genetics and Breeding Ministry of EducationRice Research Institute Shenyang Agricultural University

Plant height and tillering are crucial factors determining rice plant architecture and influencing rice grain production. In this study, multi-tillering dwarf1(mtd1), a stable multi-tiller and dwarf mutant, was screened from the ethylmethane sulfonate-treated japonica rice variety Wuyunging7. Compared with the wild type, mtd1 mutant exhibited pleiotropic phenotypes, including dwarfism, more tillers, brittle culms and delayed heading *** employing map-based cloning strategy, the gene MTD1 was finally mapped to an approximately 66-kb region on the short arm of chromosome 9. Sequencing results showed that the gene LOC_Os09g02650(BC12) in mtd1 mutant had a single nucleotide substitution(G to A), which generated a premature translation stop. Over-expressing MTD1/BC12 coding sequence rescued all the phenotypes of mtd1 mutants including plant height and tillers, which confirms that BC12 is the mutated gene in mtd1 *** reverse transcription-PCR analysis showed that MTD1/BC12 could negatively regulate the expression of MONOCULM 1, IDEAL PLANT ARCHITECTURE1 and Tillering and Dwarf 1, and control rice tillering. Remarkably, a-amylase activity analysis and gibberellic acid(GA)treatment showed that the dwarf phenotype of mtd1 mutant was dependent on GA biosynthesis pathway. These results facilitated to further uncover the molecular mechanism of the growth and development in rice.

关键词：等位基因分蘖水稻株高抽头延迟线矮秆突变体甲基磺酸乙酯生物合成途径

Characterization and fine mapping of an early senescence mutant (es-t) in Oryza sativa L.

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Chinese Science Bulletin 2011年第23期56卷 2437-2443页

作者： YANG YaoLong rao yuchun LIU HuiJuan FANG YunXia DONG GuoJun HUANG LiChao LENG YuJia GUO LonBiao ZHANG GuangHeng HU Jiang GAO ZhenYu QIAN Qian ZENG DaLi State Key Laboratory of Rice Biology China National Rice Research Institute Hangzhou 310006 China College of Life and Environment Sciences Hangzhou Normal University Hangzhou 310036 China

An es-t (early senescence-temporary) mutant, produced by ethylene methylsulfonate treatment of strain Nipponbare, was identified in rice. The leaves of es-t appeared yellow at the seedling stage, and had decreased chlorophyll content. Rust spots were found during growth in es-t, especially at the leaf margin and tip. The plants showed a typical early-senescence phenotype at the milky stage. The leaf surface of es-t appeared smoother than wild-type leaves under a scanning electron microscope, because the leaves lack siliceous protuberances around the stoma. Chloroplasts grow abnormally and are filled with many starch grains in es-t. Paraffin section analysis showed that the development of the sclerenchyma cells and vascular bundles were also abnormal in es-t. Genetic analysis indicated that es-t was controlled by a recessive gene, which was finely mapped to a 42-kb interval on chromosome 5. These results will facilitate the positional cloning and functional studies of the gene.

关键词： ES 衰老早期精细定位突变水稻扫描电子显微镜表征

The pleiotropic ABNORMAL FLOWER AND DWARF1 affects plant height, floral development and grain yield in rice

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Journal of Integrative Plant Biology 2016年第6期58卷 529-539页

作者： Deyong Ren yuchun rao Liwen Wu Qiankun Xu Zizhuang Li Haiping Yu Yu Zhang Yujia Leng Jiang Hu Li Zhu Zhenyu Gao Guojun Dong Guangheng Zhang Longbiao Guo Dali Zeng Qian Qian State Key Lab for Rice Biology China National Rice Research Institute College of Chemistry and Life Sciences Zhejiang Normal University College of Life and Environmental Sciences Hangzhou Normal University

Moderate plant height and successful establishment of reproductive organs play pivotal roles in rice grain production. The molecular mechanism that controls the two aspects remains unclear in rice. In the present study,we characterized a rice gene, ABNORMAL FLOWER AND DWARF1（AFD1） that determined plant height, floral development and grain yield. The afd1 mutant showed variable defects including the dwarfism, long panicle, low seed setting and reduced grain yield. In addition, abnormal floral organs were also observed in the afd1 mutant including slender and thick hulls, and hull-like ***1 encoded a DUF640 domain protein and was expressed in all tested tissues and organs. Subcellular localization showed AFD1-green fluorescent fusion protein（GFP） was localized in the nucleus. Meantime, our results suggested that AFD1 regulated the expression of cell division and expansion related genes.

关键词： Plant height hull lodicule grain yield AFD1 rice(Oryza sativa)

A New-Nipponbare Rice Germplasm with High Seed-Setting Rate

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Journal of Genetics and Genomics 2014年第10期41卷 549-552页

作者： Jiang Hu Guojun Dong Yunxia Fang yuchun rao Jie Xu Dawei Xue Haiping Yu Changwei Ge Zhenyuan Shi Jiangjie Pan Li Zhu Dali Zeng Guangheng Zhang Longbiao Guo Qian Qian State Key Laboratory of Rice Biology China National Rice Research Institute College of Chemistry and Life Sciences Zhejiang Normal University College of Life and Environment Sciences Hangzhou Normal University

Rice （Oryza sativa L.） is an important crop providing staple food for more than half the world＇s population and also considered as a model plant for molecular biological study of the cereals. In 1998, the large-scale sequencing of japonica rice cultivar Nipponbare （bred at Aichi Agricultural Center in Japan and released in 1963） was initiated by International Rice Genome Sequencing Project （IRGSP） and the high-quality draft of genome was announced in 2002 （Goff et al,, 2002）. Owing to its easy genetic transformation and released whole genome sequencing data, Nipponbare is widely used in functional genomic research （Piffanelli et al., 2007; Luan et al., 2008; Hu et al., 2010; Thang et al., 2010; Tabuchi et al., 2011; Zhang et al., 2011; Jiang et al., 2012; Lu et al.. 2013）.

关键词： length In more A New-Nipponbare Rice Germplasm with High Seed-Setting Rate than

基于CRISPR-Cas9基因编辑技术在作物中的应用

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生物工程学报 2023年第2期39卷 399-424页

作者：殷文晶陈振概黄佳慧叶涵斐芦涛路梅饶玉春浙江师范大学生命科学学院浙江金华321004

CRISPR-Cas9系统是由成规律的间隔短回文重复序列(clustered regularly interspaced short palindromic repeat,CRISPR)及其相关蛋白构成,广泛存在于细菌和古细菌中,该系统作为一种特异性免疫保护,可抵御病毒、噬菌体的二次侵染。CRISPR... 详细信息

CRISPR-Cas9系统是由成规律的间隔短回文重复序列(clustered regularly interspaced short palindromic repeat,CRISPR)及其相关蛋白构成,广泛存在于细菌和古细菌中,该系统作为一种特异性免疫保护,可抵御病毒、噬菌体的二次侵染。CRISPR-Cas9技术是继锌指核酸酶(zinc finger nucleases,ZFNs)和类转录激活因子效应物核酸酶(transcription activator like effector nucleases,TALENs)技术之后的第三代新型靶向基因组编辑技术。CRISPR-Cas9技术具有灵活、高效、生产成本低且特异性高等优点,极大地促进了基因功能方面的研究。CRISPR-Cas9技术经过不断的完善和发展,目前该技术已广泛应用在各个领域。本文介绍了CRISPR-Cas9技术的产生、工作机制及优势;综述了CRISPR-Cas9技术在基因敲除、基因敲入、基因调控和基因组方面的应用以及在水稻、小麦、玉米、大豆和马铃薯等重要粮食作物以及对作物驯化的应用;分析了CRISPR-Cas9技术目前面临的问题和挑战,并展望了CRISPR-Cas9技术的发展及应用前景。

关键词： CRISPR-Cas9技术基因编辑基因工程水稻作物

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解读生物学选考中的细胞周期同步化

生物学教学 2023年第7期48卷 85-87页

作者：陈昶旭沈思怡饶玉春浙江省青田县中学丽水323900 浙江师范大学教师教育学院金华321004

本文对浙江选考中涉及的细胞周期同步化的常用方法进行解读,包括营养限制性培养、抑制CDK-cyclin复合体活性、抑制纺锤体形成和抑制DNA合成,以期为高中生物学教师教学提供参考。

关键词：细胞周期同步化细胞周期检查点生物学选考

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水稻卷叶细胞学与分子机制研究进展

植物学报 2018年第6期53卷 848-855页

作者：周亭亭饶玉春任德勇浙江师范大学化学与生命科学学院金华321004 中国水稻研究所水稻生物学国家重点实验室杭州310006

卷叶性状是水稻(Oryza sativa)重要的育种指标之一。研究表明,水稻叶片适度卷曲对植株的光合作用、株型及增产等均具有重要作用。该文综述了水稻叶片卷曲的相关研究进展,对水稻叶片卷曲的细胞学形成机制和相关调控基因的分子机制进行了... 详细信息

卷叶性状是水稻(Oryza sativa)重要的育种指标之一。研究表明,水稻叶片适度卷曲对植株的光合作用、株型及增产等均具有重要作用。该文综述了水稻叶片卷曲的相关研究进展,对水稻叶片卷曲的细胞学形成机制和相关调控基因的分子机制进行了阐述,以期深入阐明水稻叶片卷曲的细胞学和分子机制,促进卷叶性状在水稻育种中的应用。

关键词：水稻卷叶细胞学机制分子机制