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Targeted mutagenesis of POLYAMINE OXIDASE 5 that negatively regulates mesocotyl elongation enables the generation of direct-seeding rice with improved grain yield

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Molecular Plant 2021年第2期14卷 344-351页

作者： Yusong Lv gaoneng shao Guiai Jiao Zhonghua Sheng Lihong Xie Shikai Hu shaoqing Tang Xiangjin Wei Peisong Hu State Key Laboratory of Rice Biology China National Center for Rice ImprovementChina National Rice Research InstituteHangzhou 310006China National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070China

Under conditions of labor or resource scarcity,direct seeding,rather than transplantation,is the preferred mode of rice(Oryza sativa)*** approach requires varieties that exhibit uniform seedling *** elongation(ME),the main driver of rapid emergence of rice seedlings from soil,is enhanced by darkness and inhibited by *** polyamine oxidases(PAOs)oxidize polyamines(PAs)and release H2O2,Here,we established that OsPAO5 expression in rice seedlings is increased in the presence of light and inhibited by *** determine its role in ME,we created OsPAO5 mutants using CRISPR/*** with the wild type,pao5 mutants had longer mesocotyls,released less H2O2,and synthesized more *** mutant seedlings emerged at a higher and more uniform rate,indicating their potential for use in direct *** polymorphism analysis revealed that an SNP(PAO5-578G/A)located 578 bp upstream of the OsPAO5 start codon alters its expression,and was selected during rice mesocotyl *** PAO5-578G genotype conferring a long mesocotyl mainly exists in wild rice,most Aus accessions,and some Geng(Japonica)***,knocking out OsPAO5 can remarkably increase the grain weight,grain number,and yield *** summary,we developed a novel strategy to obtain elite rice with higher emergence vigor and yield potential,which can be conveniently and widely used to breed varieties of direct-seeding rice.

关键词： Oryza sativa direct seeding mesocotyl elongation grain yield polyamine oxidase 5

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GRAIN INCOMPLETE FILLING 2 regulates grain filling and starch synthesis during rice caryopsis development

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Journal of Integrative Plant Biology 2017年第2期59卷 134-153页

作者： Xiangjin Wei Guiai Jiao Haiyan Lin Zhonghua Sheng gaoneng shao Lihong Xie shaoqing Tang Qingguo Xu Peisong Hu State Key Laboratory of Rice Biology Key Laboratory of Rice Biology and Breeding of Ministry of Agriculture China National Rice Research Institute Hangzhou 310006 China . College of Agronomy Hunan Agricultural University Changsha 410128 China Agricultural Genomes Institute at Shenzhen Chinese Academy of Agricultural Sciences Shenzhen 518120 China

Rice grain filling determines grain weight, final yield and grain quality. Here, a rice defective grain filling mutant, gif2, was identified. Grains ofgif2 showed a slower filling rate and a significant lower final grain weight and yield compared to wild-type. The starch content in gilt2 was noticeably decreased and its physicochemical properties were also altered. Moreover, gif2 endosperm cells showed obvious defects in compound granule formation. Posi- tional cloning identified GIF2 to encode an ADP-glucose pyrophosphorylase （AGP） large subunit, AGPL2; consequently, AGP enzyme activity in gif2 endosperms was remarkably decreased. GIF2 is mainly expressed in developing grains and the coded protein localizes in the cytosol. Yeast two hybrid assay showed that GIF2 interacted with AGP small subunits OsAGPS% OsAGPS2a and OsAGPS2b. Transcript levels for granule-bound starch synthase, starch synthase, starch branching enzyme and starch debranching enzyme were distinctly elevated in gif2 grains. In addition, the level of nucleotide diversity of the GIF2 locus was extremely low in both cultivated and wild rice. All of these results suggest that GIF2 plays important roles in the regulation of grain filling and starch biosynthesis during caryopsis development, and that it has been preserved during selection throughout domestication of modern rice.

关键词： GIF GRAIN INCOMPLETE FILLING 2 regulates grain filling and starch synthesis during rice caryopsis development Figure AGPS RNAi

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Mapping of qGL7-2, a grain length QTL on chromosome 7 of rice

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Journal of Genetics and Genomics 2010年第8期37卷 523-531页

作者： gaoneng shao shaoqing Tang Ju Luo Guiai Jiao Xiangjin Wei Ao Tang Jianli Wu Jieyun Zhuang Peisong Hu State Key Laboratory of Rice Biology China National Rice Research lnstitt~te Hangzhou 310006 China

A residual heterozygous line （RHL） carrying a heterozygous segment between two S SR loci RM 11 and RM134 on the rice chromo- some 7 was selected from a set of recombinant inbred lines from the cross D50 （javanica）/HB277 （indica）. The former parent produces much longer grains than the latter. Selfed progenies of this selection were analyzed genotypically （SSRs） and phenotypically （grain length）. Grain length was discontinuously variable in the mapping populations, allowing for the placement of this QTL qGL7-2 within a -4.8 cM interval defined by RM351 and RM234. A set of new markers within this region were developed, which narrowed the QTL to a 278 kb region defined by the markers Indell and RM21945. This region contains 49 predicted genes. The results also suggest that the novel allele for grain length will be used for the application of marker assisted selection for the improvement of grain length.

关键词： grain length mapping quantitative trait locus residual heterozygous line rice

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Improved Eating and Cooking Quality of indica Rice Cultivar YK17 via Adenine Base Editing of Wx^(a)Allele of Granule-Bound Starch Synthase I(GBSS I)

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Rice science 2021年第5期28卷 427-430,I0010-I0017页

作者： Mahmuda Binte Monsur Cao Ni Wei Xiangjin Xie Lihong Jiao Guiai TAng shaoqing Nese Sreeniwsulu shao gaoneng Hu Peisong State Key Laboratory of Rice Biology China National Rice Research InstituteHangzhou 310006China Applied Functional Genomics Cluster Grain Quality and Nutrition CentreStrategic innovation PlatformInternational Rice Research InstituteLos Banos 4030the Philippines

Amylose content(AC)is the key determinant of eating and cooking quality(ECQ)of *** major Wxa allele of granule-bound starch synthase I(GBSS I)in indica rice produces higher AC,making rice hard and dry after *** work has improved ECQ of japonica rice via clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9)or cytosine base editing(CBE)techniques.

关键词： CRISPR/Cas9 cooking Starch

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CDE4 encodes a pentatricopeptide repeat protein involved in chloroplast RNA splicing and affects chloroplast development under low-temperature conditions in rice

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Journal of Integrative Plant Biology 2021年第10期63卷 1724-1739页

作者： Xinyong Liu Xichun Zhang Ruijie Cao Guiai Jiao Shikai Hu gaoneng shao Zhonghua Sheng Lihong Xie shaoqing Tang Xiangjin Wei Peisong Hu State Key Laboratory of Rice Biology China National Center for Rice ImprovementChina National Rice Research InstituteHangzhou 310006China Guizhou Rice Research Institute Guizhou Academy of Agricultural SciencesGuiyang 550006China

Pentatricopeptide repeat(PPR)proteins play important roles in the post-transcriptional modification of organellar RNAs in ***,the function of most PPR proteins remains ***,we characterized the rice(Oryza sativa L.)chlorophyll deficient 4(cde4)mutant which exhibits an albino phenotype during early leaf development,with decreased chlorophyll contents and abnormal chloroplasts at low-temperature(20℃).Positional cloning revealed that CDE4 encodes a P-type PPR protein localized in *** the cde4 mutant,plastid-encoded polymerase(PEP)-dependent transcript levels were significantly reduced,but transcript levels of nuclear-encoded genes were increased compared to wild-type plants at 20℃.CDE4 directly binds to the transcripts of the chloroplast genes rpl2,ndhA,and *** splicing of these transcripts was defective in the cde4 mutant at 20℃,but was normal at 32℃.Moreover,CDE4 interacts with the guanylate kinase VIRESCENT 2(V2);overexpression of V2 enhanced CDE4 protein stability,thereby rescuing the cde4 phenotype at 20℃.Our results suggest that CDE4 participates in plastid RNA splicing and plays an important role in rice chloroplast development under lowtemperature conditions.

关键词： CDE4 chloroplast development pentatricopeptide repeat(PPR)protein RNA splicing low-temperature Oryza sativa L.

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MONOCULM 3,an Ortholog of WUSCHEL in Rice,Is Required for Tiller Bud Formation

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Journal of Genetics and Genomics 2015年第2期42卷 71-78页

作者： Zefu Lu gaoneng shao Jinsong Xiong Yongqing Jiao Jing Wang Guifu Liu Xiangbing Meng Yan Liang Guosheng Xiong Yonghong Wang Jiayang Li State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing) Institute of Genetics and Developmental BiologyChinese Academy of Sciences College of Horticulture Nanjing Agricultural University Oil Crops Research Institute China Academy of Agricultural Science Rice Research Institute Sichuan Agricultural University Agricultural Genome Institute China Academy of Agricultural Science

WUSCHEL （WUS） plays an essential role for the maintenance of meristem activity in dicots, but its function is still elusive in monocots. We isolated a new monoculm mutant, monoculm 3 （moc3）, in which a point mutation causes the premature termination of rice O. sativa WUS （OsWUS）. Morphological observation revealed that the formation of tiller buds was disrupted in moc3. MOC3 was localized in the nuclear and could interact with TOPLESS-RELATED PROTEINS （TPRs）. The expression of MOC3 was induced by cytokinins and defection of MOC3 affected the expression of several two-component cytokinin response regulators, OsRRs and ORRs. Our results suggest that MOC3 is required for the formation of axillary buds and has a complex relationship with cytokinins.

关键词： MOC3 WUSCHEL Tiller bud formation Cytokinins Oryza sativa L

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OsVP1 activates Sdr4 expression to control rice seed dormancy via the ABA signaling pathway

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The Crop Journal 2021年第1期9卷 68-78页

作者： Wenqiang Chen Wei Wang Yusong Lyu Yawen Wu Pingliang Huang Shikai Hu Xiangjin Wei Guiai Jiao Zhonghua Sheng shaoqing Tang gaoneng shao Ju Luo State Key Laboratory of Rice Biology China National Rice Research InstituteHangzhou 310006ZhejiangChina Institute of Crop Science and Nuclear Technology Utilization Zhejiang Academy of Agricultural SciencesHangzhou 310021ZhejiangChina

Pre-harvest sprouting(PHS)is a disadvantageous trait in cereal production worldwide,causing large economic losses each *** regulation mechanism is still *** generated the Oryza sativa Viviparous1(OsVP1)mutant using gene editing technique,which shows increased PHS compared with that of the wild type ***1 is localized mainly in the nucleus and expressed in various tissues and *** of Seed dormancy 4(Sdr4),a key gene controlling PHS,was sharply reduced in OsVP1 ***1 bound to the specific motif CACCTG in the promoter of Sdr4 and activated its expression in rice *** of Sdr4 reduced the high seed germination rate of OsVP1 mutant cr-osvp1-1,showing that Sdr4 acts as a downstream target of *** OsVP1 and Sdr4 loss-of-function mutants were insensitive to exogenous ABA and employed the ABA signaling pathway in regulating seed *** findings shed light on the control of seed dormancy aimed at preventing PHS in rice.

关键词： Rice Pre-harvest sprouting Seed dormancy OsVP1 Sdr4 ABA

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Editing of Rice Endosperm Plastidial Phosphorylase Gene OsPho1 Advances Its Function in Starch Synthesis

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Rice science 2021年第3期28卷 209-211页

作者： LIU Song shao gaoneng JIAO Guiai ZHU Maodi WU Jiamin CAO Ruijie CHEN Yujuan XIE Lihong SHENG Zhonghua TANG shaoqing China National Rice Research Institute Hangzhou 310006China

OsPho1 in Zhonghua 11(ZH11)was edited using the clustered regularly interspaced short palindromic repeatsassociated endonuclease 9(CRISPR/Cas9)*** homozygous T1 mutants(cr-pho1-34 and cr-pho1-37)displayed a chalky endosperm with a white core,which significantly decreased 1000-grain *** addition,many rounded starch granules and abnormal amyloplasts were present in the central region of mutant endosperm cells with increased amylose and lipid contents,decreased total protein content,and altered physicochemical properties of *** OsPho1 protein is localized in chloroplasts,and quantitative real-time PCR(qRT-PCR)andβ-glucuronidase(GUS)staining indicated that OsPho1 was highly expressed in seeds at 5 d after fertilization(DAF).OsPho1 mutations displayed close relationships with plastidial phosphoglucomutase and ADPGlc pyrophosphorylase based onα-D-glucose-1P at different ***,the expressions of starch metabolismrelated genes were also altered in the mutant,and the overexpression of OsPho1 may cause grain chalkiness.

关键词： CRISPR/Cas9 Pho Starch

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An orchestrated ethylene-gibberellin signaling cascade contributes to mesocotyl elongation and emergence of rice direct seeding

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Journal of Integrative Plant Biology 2024年第7期66卷 1427-1439页

作者： Yusong Lyu Xinli Dong Shipeng Niu Ruijie Cao gaoneng shao Zhonghua Sheng Guiai Jiao Lihong Xie Shikai Hu shaoqing Tang Xiangjin Wei Peisong Hu State Key Laboratory of Rice Biology and Breeding China National Center for Rice ImprovementChina National Rice Research InstituteHangzhou 310006China College of Agriculture and Biotechnology Zhejiang UniversityHangzhou 310058China

A mechanized direct seeding of rice with less labor and water usage,has been widely ***,this approach requires varieties that exhibit uniform seedling *** elongation(ME)offers the main drive of fast emergence of rice seedlings from soils;nevertheless,its genetic basis remains ***,we identify a major rice quantitative trait locus Mesocotyl Elongation1(qME1),an allele of the Green Revolution gene Semi-Dwarf1(SD1),encoding GA20-oxidase for gibberellin(GA)***1 expression is strongly induced by soil depth and *** rice grains are direct-seeded in soils,the ethylene core signaling factor OsEIL1 directly promotes ME1 transcription,accelerating bioactive GA *** GAs further degrade the DELLA protein SLENDER RICE 1(SLR1),alleviating its inhibition of rice PHYTOCHROME-INTERACTING FACTOR-LIKE13(OsPIL13)to activate the downstream expansion gene OsEXPA4 and ultimately promote rice seedling ME and *** ancient traits of long mesocotyl and strong emergence ability in wild rice and landrace were gradually lost in company with the Green Revolution dwarf breeding process,and an elite ME1-R allele(D349H)is found in some modern Geng varieties(long mesocotyl lengths)in northern China,which can be used in the direct seeding and dwarf breeding of Geng ***,the ectopic and high expression of ME1 driven by mesocotyl-specific promoters resulted in rice plants that could be direct-seeded without obvious plant architecture or yield ***,we reveal the molecular mechanism of rice ME,and provide useful information for breeding new Green Revolution varieties with long mesocotyl suitable for direct-seeding practice.

关键词： rice(Oryza sativa L.) direct seeding Green Revolu-tion mesocotyl elongation ethylene gibberellin semi-dwarf1(SD1)

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Integrated transcriptome, small RNA, and degradome analysis to elucidate the regulation of rice seedling mesocotyl development during the passage from darkness to light

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The Crop Journal 2020年第6期8卷 918-928页

作者： Yusong Lyu Xiangjin Wei Min Zhong Shipeng Niu Shakeel Ahmad gaoneng shao Guiai Jiao Zhonghua Sheng Lihong Xie Shikai Hu Yawen Wu shaoqing Tang Peisong Hu State Key Laboratory of Rice Biology China National Center for Rice ImprovementChina National Rice Research InstituteHangzhou 310006ZhejiangChina National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070HubeiChina

The mesocotyl,a structure located between the basal part of the seminal root and the coleoptile node of seedlings,contributes to pushing the shoot tip through the soil surface,a function that is essential for the uniform emergence of direct-seeded *** elongation is inhibited by light and induced in *** investigation of an indica rice(P25)with vigorous mesocotyl elongation was aimed at identifying the"omics"basis of its lightinduced growth inhibition.A transcriptomic comparison between mesocotyl tissues that had developed in the dark and then been exposed to light identified many differentially expressed genes(DEGs)and differentially abundant micro RNAs(mi RNAs).Degradome sequencing analysis revealed 27 negative mi RNA-target pairs.A co-expression regulatory network was constructed based on the mi RNAs,their corresponding targets,and DEGs with a common Gene Ontology *** suggested that auxin and light,probably antagonistically,affect mesocotyl elongation by regulating polyamine oxidase activity.

关键词： Oryza sativa L. Mesocotyl Transcriptome MicroRNAome Degradome

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