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OsHemA gene, encoding glutamyl-tRNA reductase(GluTR) is essential for chlorophyll biosynthesis in rice(Oryza sativa)

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Journal of Integrative agriculture 2020年第3期19卷 612-623页

作者： ZENG Zhao-qiong LIN Tian-zi ZHAO Jie-yu ZHENG Tian-hui XU Le-feng WANG Yi-hua LIU Ling-long JIANG Ling CHEN Sai-hua WAN Jian-min state key laboratory for crop genetics&germplasm enhancement/jiangsu provincial center of plant gene engineering/college of agriculture Nanjing Agricultural UniversityNanjing 210095P.R.China Nanchong Academy of Agricultural Sciences Nanchong 637000P.R.China key laboratory of plant Functional Genomics Ministry of Education/CoInnovation Center for Modern Production Technology of Grain CropsYangzhou UniversityYangzhou 225009P.R.China

Chlorophyll(Chl) biosynthesis is essential for photosynthesis and plant ***-tRNA reductase(GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde(GSA) and initiates the chlorophyll *** though the main role of GluTR has been established,the effects caused by natural variations in its corresponding gene remain largely ***,we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency,designated as *** intact thylakoid lamellar structure,the cbd1 plant showed light green leaves and reduced Chl and carotenoids(Cars) content significantly compared to the wild *** map-based gene cloning,the mutation was restricted within a 57-kb region on chromosome 10,in which an mPingA miniature inverted-repeat transposable element(MITE) inserted in the promoter region of OsHemA *** leaf color and the pigment contents in cbd1 were recovered in a complementation test,confirming OsHemA was responsible for the mutant *** was uniquely predicted to encode GluTR and its expression level was dramatically repressed in *** transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus.A majority of Chl biosynthesis genes,except for POR and CHLG,were down-regulated synchronously by the repression of OsHemA,suggesting that an attenuation occurred in the Chl biosynthesis ***,we found major agronomic traits involved in rice yield were statistically unaffected,except for the number of full grains per panicle was increased in ***,OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.

关键词： OsHemA GluTR chlorophyll biosynthesis

Young Leaf White Stripe encodes a P-type PPR protein required for chloroplast development

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Journal of Integrative plant Biology 2023年第7期65卷 1687-1702页

作者： Jie Lan Qibing Lin Chunlei Zhou Xi Liu Rong Miao Tengfei Ma Yaping Chen Changling Mou Ruonan Jing Miao Feng Thanhliem Nguyen Yulong Ren Zhijun Cheng Xin Zhang Shijia Liu Ling Jiang Jianmin Wan state key laboratory of crop genetics and germplasm enhancement Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjing 210095China National key Facility for crop gene Resources and genetic Improvement Institute of Crop ScienceChinese Academy of Agricultural SciencesBeijing 100081China

Pentatricopeptide repeat(PPR) proteins function in post-transcriptional regulation of organellar gene expression. Although several PPR proteins are known to function in chloroplast development in rice(Oryza sativa), the detailed molecular functions of many PPR proteins remain ***, we characterized a rice young leaf white stripe(ylws) mutant, which has defective chloroplast development during early seedling ***-based cloning revealed that YLWS encodes a novel P-type chloroplast-targeted PPR protein with 11 PPR motifs. Further expression analyses showed that many nuclear-and plastid-encoded genes in the ylws mutant were significantly changed at the RNA and protein levels. The ylws mutant was impaired in chloroplast ribosome biogenesis and chloroplast development under low-temperature conditions. The ylws mutation causes defects in the splicing of atpF, ndhA, rpl2,and rps12, and editing of ndhA, ndhB, and rps14transcripts. YLWS directly binds to specific sites in the atpF, ndhA, and rpl2 pre-mRNAs. Our results suggest that YLWS participates in chloroplast RNA group II intron splicing and plays an important role in chloroplast development during early leaf development.

关键词： chloroplast development PPR protein ribosome biogenesis RNA splicing

Rice FLOURY ENDOSPERM 18 encodes a pentatricopeptide repeat protein required for 5′ processing of mitochondrial nad5 messenger RNA and endosperm development

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Journal of Integrative plant Biology 2021年第5期63卷 834-847页

作者： Mingzhou Yu Mingming Wu Yulong Ren Yihua Wang Jingfang Li Cailin Lei Yinglun Sun Xiuhao Bao Hongming Wu Hang Yang Tian Pan Yongfei Wang Ruonan Jing Mengyuan Yan Houda Zhang Lei Zhao Zhichao Zhao Xin Zhang Xiuping Guo Zhijun Cheng Bing Yang Ling Jiang Jianmin Wan state key laboratory for crop genetics and germplasm enhancement Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjing 210095China National key Facility for crop gene Resources and genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China

Pentatricopeptide repeat(PPR) proteins, composing one of the largest protein families in plants,are involved in RNA binding and regulation of organelle RNA metabolism at the posttranscriptional level. Although several PPR proteins have been implicated in endosperm development in rice(Oryza sativa), the molecular functions of many PPRs remain obscure. Here, we identified a rice endosperm mutant named floury endosperm 18(flo18) with pleiotropic defects in both reproductive and vegetative ***-based cloning and complementation tests showed that FLO18 encodes a mitochondriontargeted P-type PPR protein with 15 PPR *** function was disrupted in the flo18 mutant, as evidenced by decreased assembly of Complex I in the mitochondrial electron transport chain and altered mitochondrial morphology. Loss of FLO18 function resulted in defective 5′-end processing of mitochondrial nad5 transcripts encoding subunit 5 of nicotinamide adenine dinucleotide hydrogenase. These results suggested that FLO18 is involved in 5′-end processing of nad5 messenger RNA and plays an important role in mitochondrial function and endosperm development.

关键词： aleurone layer endosperm development mitochondria PPR nad5,5′end processing

ESCRT-Ⅲ component OsSNF7.2 modulates leaf rolling by trafficking and endosomal degradation of auxin biosynthetic enzyme OsYUC8 in rice

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Journal of Integrative plant Biology 2023年第6期65卷 1408-1422页

作者： Liang Zhou Saihua Chen Maohong Cai Song Cui Yulong Ren Xinyue Zhang Tianzhen Liu Chunlei Zhou Xin Jin Limin Zhang Minxi Wu Shuyi Zhang Zhijun Cheng Xin Zhang Cailin Lei Qibing Lin Xiuping Guo Jie Wang Zhichao Zhao Ling Jiang Shanshan Zhu Jianmin Wan state key laboratory for crop genetics and germplasm enhancement Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjing 210095China National key Facility for crop gene Resources and genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China jiangsu key laboratory of crop Genomics and Molecular Breeding/key laboratory of plant Functional Genomics of the Ministry of Education/jiangsu key laboratory of crop genetics and Physiology Agricultural College of Yangzhou UniversityYangzhou 225009China

The endosomal sorting complex required for transport(ESCRT)is highly conserved in eukaryotic cells and plays an essential role in the biogenesis of multivesicular bodies and cargo degradation to the plant vacuole or *** ESCRT components affect a variety of plant growth and development processes,their impact on leaf development is rarely ***,we found that OsSNF7.2,an ESCRT-Ⅲ component,controls leaf rolling in rice(Oryza sativa).The Ossnf7.2 mutant rolled leaf 17(rl17)has adaxially rolled leaves due to the decreased number and size of the bulliform ***7.2is expressed ubiquitously in all tissues,and its protein is localized in the endosomal ***7.2 homologs,including OsSNF7,OsSNF7.3,and OsSNF7.4,can physically interact with OsSNF7.2,but their single mutation did not result in leaf *** ESCRT complex subunits,namely OsVPS20,OsVPS24,and OsBRO1,also interact with *** assays revealed that OsSNF7.2 interacts with OsYUC8 and aids its vacuolar *** Osyuc8and rl17 Osyuc8 showed rolled leaves,indicating that OsYUC8 and OsSNF7.2 function in the same pathway,conferring leaf *** study reveals a new biological function for the ESCRT-Ⅲcomponents,and provides new insights into the molecular mechanisms underlying leaf rolling.

关键词： bulliform cell leaf rolling OsSNF7.2 OsYUC8 rice(Oryza sativa) vacuolar degradation

Molecular Cloning and Characterization of an Allene Oxide Cyclase gene Associated with Fiber Strength in Cotton

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Journal of Integrative agriculture 2014年第10期13卷 2113-2121页

作者： WANG Li-man ZHU You-min TONG Xiang-chao HU Wen-jing CAI Cai-ping GUO Wang-zhen state key laboratory of crop genetics & germplasm enhancement Hybrid Cotton R&D Engineering Research CenterMinistry of Education/College of AgricultureNanjing Agricultural University

Allene oxide cyclase （AOC） is one of the most important enzymes in the biosynthetic pathway of the plant hormone jasmonic acid （JA）. AOC catalyzes the conversion ofallene oxide into 12-oxo-phytodienoic acid （OPDA）, a precursor of JA. Using 28K cotton genome array hybridization, an expressed sequence tag （EST; GenBank accession no. ES792958） was investigated that exhibited significant expression differences between lintless-fuzzless XinWX and linted-fuzzless XinFLM isogenic lines during fiber initiation stages. The EST was used to search the Gossypium EST database （http：//***/） for corresponding cDNA sequences encoding full-length open reading frames （ORFs）. Identified ORFs were confirmed using transcriptional and genomic data. As a result, a novel gene encoding AOC in cotton （Gossypium hirsutum AOC; GenBank accession no. KF383427） was cloned and characterized. The 741-bp GhAOC gene comprises three exons and two introns and encodes a polypeptide of 246 amino acids. Two homologous copies were identified in the tetraploid cotton species G. hirsutum acc. TM-1 and G. barbadense cv. Hai7124, and one copy in the diploid cotton species G. herbaceum and G. raimondii, qRT-PCR showed that the GhAOC transcript was abundant in cotton fiber tissues from 8 to 23 days post anthesis （DPA）, and the expression profiles were similar in the two cultivated tetraploid cotton species G. hirsutum acc. TM- 1 and G. barbadense cv. Hai7124, with a higher level of transcription in the former. One copy of GhAOC in tetraploid cotton was localized to chromosome 24 （Chr. D8） using the subgenome-specific single nucleotide polymorphism （SNP） marker analysis, which co-localized GhAOC to within 10 cM of a fiber strength quantitative trait locus （QTL） reported previously. GhAOC was highly correlated with fiber quality and strength （P=0.014） in an association analysis, suggesting a possible role in cotton fiber development, especially in secondary cell wall thickening.

关键词： allene oxide cyclase structure expression pattern fiber development association analysis cotton

Mitochondrion-targeted PENTATRICOPEPTIDE REPEAT5 is required for cis-splicing of nad4 intron 3 and endosperm development in rice

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The crop Journal 2021年第2期9卷 282-296页

作者： Long Zhang Yanzhou Qi Mingming Wu Lei Zhao Zhichao Zhao Cailin Lei Yuanyuan Hao Xiaowen Yu Yinglun Sun Xin Zhang Xiuping Guo Yulong Ren Jianmin Wan National key Facility for crop gene Resources and genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China state key laboratory for crop genetics and germplasm enhancement Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjing 210095JiangsuChina college of Bioscience and Biotechnology Yangzhou UniversityYangzhou 225009JiangsuChina

Endosperm as the storage organ of starch and protein in cereal crops largely determines grain yield and *** the fact that several pentatricopeptide repeat(PPR)proteins required for endosperm development have been identified in rice,the molecular mechanisms of many P-type PPR proteins in endosperm development remains ***,we isolated a rice floury endosperm mutant ppr5 that developed small starch grains and an abnormal aleurone layer,accompanied by decreased starch,protein,and amylose ***-based cloning combined with a complementation test demonstrated that PPR5 encodes a P-type PPR protein that is localized to the *** mutation in PPR5 caused reduced splicing efficiency of mitochondrial NADH dehydrogenase 4(nad4)gene intron 3 and reduced complex I assembly and *** of PPR5 function greatly upregulated expression of alternative oxidases(AOXs),reduced ATP production,and affected mitochondrial *** demonstrate that PPR5,as a P-type PPR protein,is required for mitochondrial function and endosperm development by controlling the cis-splicing of mitochondrial nad4 intron 3.

关键词： Floury endosperm PPR Mitochondria RNA splicing Oryza sativa

DHD4,a CONSTANS-like family transcription factor,delays heading date by affecting the formation of the FAC complex in rice

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Molecular plant 2021年第2期14卷 330-343页

作者： Maohong Cai Shanshan Zhu Mingming Wu Xiaoming Zheng Jiachang Wang Liang Zhou Tianhui Zheng Song Cui Shirong Zhou Chaonan Li Huan Zhang Juntao Chai Xinyue Zhang Xin Jin Zhijun Cheng Xin Zhang Cailin Lei Yulong Ren Qibing Lin Xiuping Guo Lei Zhao Jie Wang Zhichao Zhao Ling Jiang Haiyang Wang Jianmin Wan National key Facility for crop gene Resources end genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China state key laboratory for crop genetics and germplasm enhancement Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjing210095China

Heading date(or flowering time)is one of the most important agronomic traits in rice,influencing its regional adaptability and crop *** major-effect genes for rice heading date have been identified,but in practice they are difficult to be used for rice molecular breeding because of their dramatic effects on heading *** with minor effects on heading date,which are more desirable for fine-tuning flowering time without significant yield penalty,were seldom *** this study,we identified a new minor-effect heading date repressor,Delayed Heading Date 4(DHD4).The dhd4 mutant shows a slightly earlier flowering phenotype without a notable yield penalty compared with wild-type plants under natural long-day ***4 encodes a CONSTANS-like transcription factor localized in the ***,biochemical,and genetic assays show that DHD4 can compete with 14-3-3 to interact with OsFD1,thus affecting the formation of the Hd3a-14-3-3-OsFD1 triprotein FAC complex,resulting in reduced expression of OsMADS14 and OsMADS15,and ultimately delaying *** together,these results shed new light on the regulation of flowering time in rice and provide a promising target for fine-tuning flowering time to improve the regional adaptability of rice.

关键词： rice Oryza sativa delayed heading date 4 DHD4 minor-effect OsFD1 heading date

Up-regulation of a homeodomain-leucine zipper gene HD-1 contributes to trichome initiation and development in cotton

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Journal of Integrative agriculture 2019年第2期18卷 361-371页

作者： NIU Er-li CAI Cai-ping BAO Jiang-hao WU Shuang ZHAO Liang GUO Wang-zhen state key laboratory of crop genetics&germplasm enhancement Ministry of Science and Technology/Hybrid Cotton R&D Engineering Research CenterMinistry of Education/College of AgricultureNanjing Agricultural UniversityNanjing 210095P.R.China

plant trichomes originate from epidermal *** this work,we demonstrated that a homeodomain-leucine zipper(HD-Zip)gene,Gh_A06G1283(Gh HD-1A),was related to the leaf trichome trait in allotetraploid cotton and could be a candidate gene for the T_1 *** ortholog of GhHD-1A in the hairless accession Gossypium barbadense ***7124 was interrupted by a long terminal repeat(LTR)retrotransposon,while GhHD-1A worked well in the hairy accession Gossypium hirsutum *** and phylogenetic analysis showed that GhHD-1A belonged to the HD-Zip IV gene family,which mainly regulated epidermis hair development in *** of GhHD-1A and its homoeologs GhHD-1D in allotetraploid T586and Hai7124 could significantly reduce the density of leaf hairs and affect the expression levels of other genes related to leaf trichome *** analysis found that GhHD-1A mainly regulated trichome initiation on the upper epidermal hairs of leaves in cotton,while the up-regulated expression of GhHD-1A in different organs/tissues also altered epidermal trichome *** study not only helps to unravel the important roles of GhHD-1A in regulating trichome initiation in cotton,but also provides a reference for exploring the different forms of trichome development in plants.

关键词： leaf trichome map-based cloning a homeodomain-leucine zipper gene HD-1 virus-induced gene silencing (VIGS) functional differentiation

Rice FLOURY ENDOSPERM22, encoding a pentatricopeptide repeat protein, is involved in both mitochondrial RNA splicing and editing and is crucial for endosperm development

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Journal of Integrative plant Biology 2023年第3期65卷 755-771页

作者： Hang Yang Yunlong Wang Yunlu Tian Xuan Teng Zehui Lv Jie Lei Erchao Duan Hui Dong Xue Yang Yuanyan Zhang Yinglun Sun Xiaoli Chen Xiuhao Bao Rongbo Chen Chuanwei Gu Yipeng Zhang Xiaokang Jiang Wenyu Ma Pengcheng Zhang Yi Ji Yu Zhang Yihua Wang Jianmin Wan state key laboratory for crop genetics and germplasm enhancement Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjing 210095China National key Facility for crop Resources and genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China

Most of the reported P-type pentatricopeptide repeat(PPR) proteins play roles in organelle RNA stabilization and splicing. However, P-type PPRs involved in both RNA splicing and editing have rarely been reported, and their underlying mechanism remains largely unknown. Here, we report a rice floury endosperm22(flo22) mutant with delayed amyloplast development in endosperm cells. Map-based cloning and complementation tests demonstrated that FLO22 encodes a mitochondrion-localized P-type PPR *** of FLO22 resulting in defective transsplicing of mitochondrial nad1 intron 1 and perhaps causing instability of mature transcripts affected assembly and activity of complex Ⅰ, and mitochondrial morphology and function. RNA-seq analysis showed that expression levels of many genes involved in starch and sucrose metabolism were significantly down-regulated in the flo22mutant compared with the wild type, whereas genes related to oxidative phosphorylation and the tricarboxylic acid cycle were significantly upregulated. In addition to involvement in splicing as a P-type PPR protein, we found that FLO22 interacted with DYW3, a DYW-type PPR protein, and they may function synergistically in mitochondrial RNA editing. The present work indicated that FLO22 plays an important role in endosperm development and plant growth by participating in nad1 maturation and multi-site editing of mitochondrial messager RNA.

关键词： endosperm development Oryza sativa PPR proteins RNA editing RNA splicing