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Calcineurin B-like interacting protein kinase OsCIPK23 functions in pollination and drought stress responses in rice (Oryza sativa L.)

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Journal of Genetics and Genomics 2008年第9期35卷 531-543页

作者： Wenqiang Yang zhaosheng kong Edith Omo-Ikerodah Wenying Xu Qun Li Yongbiao Xue Institute of Genetics and Developmental Biology Chinese Academy of Sciences National Centre for Plant Gene Research Beijing 100190 China Graduate School of Chinese Academy of Sciences Beijing 100049 China

Drought is very harmful to grain yield due to its adverse effect on reproduction, especially on pollination process in rice. However, the molecular basis of such an effect still remains largely unknown. Here, we report the role of a member of CBL （Calcineurin B-Like） Interacting Protein Kinase （CIPK） family, OsCIPK23, in pollination and stress responses in rice. Molecular analyses revealed that it is mainly expressed in pistil and anther but up-regulated by pollination, as well as by treatments of various abiotic stresses and phytohormones. RNA interference-mediated suppression of OsCIPK23 expression significantly reduced seed set and conferred a hypersensitive response to drought stress, indicating its possible roles in pollination and drought stress. In consistent, overexpression of OsCIPK23 induced the expression of several drought tolerance related genes. Taken together, these results indicate that OsCIPK23 is a multistress induced gene and likely mediates a signaling pathway commonly shared by both pollination and drought stress responses in rice.

关键词： OsCIPK23 water pollination drought rice （Oryza sativa L.）

The two domains of cotton WLIM1a protein are functionally divergent

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Science China(Life Sciences) 2016年第2期59卷 206-212页

作者： Libo Han Yuanbao Li Yongduo Sun Haiyun Wang zhaosheng kong Guixian Xia State Key Laboratory of Plant Genomics Institute of Microbiology Chinese Academy of Sciences College of Life Sciences University of Chinese Academy of Sciences

Our previous study demonstrated that WLIM1a has dual roles in fiber elongation and secondary cell wall synthesis in upland cotton,and the protein acts either as an actin-binding protein or as a transcription factor.Because WLIM1a consists of two different LIM domains,it is possible that these elements contribute differentially to the dual functions of the protein.In this study,we dissected the two LIM domains and characterized their biochemical functions.By using red fluorescent protein(RFP)fusion,co-sedimentation,and DNA binding methods,we found that the two domains of WLIM1a,domain1(D1)and domain2(D2),possessed different biochemical properties.While D1 contributed primarily to the actin filament-bundling activity of WLIM1a,D2 contributed to the DNA-binding activity of the protein;both D1 and D2 relied on a linker sequence for their activities.In addition,we found that WLIM1a and its two LIM domains form dimers in vitro.These results may lead to a better understanding of the molecular mechanisms of dual functions of WLIM1a during cotton fiber development.

关键词： a蛋白肌动蛋白结合蛋白 LIM结构域 DNA结合活性棉花红色荧光蛋白棉纤维发育纤维伸长

Rhizobial infection triggers systemic transport of endogenous RNAs between shoots and roots in soybean

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Science China(Life Sciences) 2020年第8期63卷 1213-1226页

作者： Chen Zhang Meifang Qi Xiaxia Zhang Qi Wang Yanjun Yu Yijing Zhang zhaosheng kong State Key Laboratory of Plant Genomics Institute of MicrobiologyInnovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China Shanghai Institute of Plant Physiology and Ecology Chinese Academy of SciencesShanghai 200032China University of Chinese Academy of Sciences Beijing 100049China

Legumes have evolved a symbiotic relationship with rhizobial bacteria and their roots form unique nitrogen-fixing organs called nodules.Studies have shown that abiotic and biotic stresses alter the profile of gene expression and transcript mobility in plants.However,little is known about the systemic transport of RNA between roots and shoots in response to rhizobial infection on a genome-wide scale during the formation of legume-rhizobia symbiosis.In our study,we found that two soybean(Glycine max)cultivars,Peking and Williams,show a high frequency of single nucleotide polymorphisms;this allowed us to characterize the origin and mobility of transcripts in hetero-grafts of these two cultivars.We identified 4,552 genes that produce mobile RNAs in soybean,and found that rhizobial infection triggers mass transport of m RNAs between shoots and roots at the early stage of nodulation.The majority of these mRNAs are of relatively low abundance and their transport occurs in a selective manner in soybean plants.Notably,the mRNAs that moved from shoots to roots at the early stage of nodulation were enriched in many nodule-related responsive processes.Moreover,the transcripts of many known symbiosis-related genes that are induced by rhizobial infection can move between shoots and roots.Our findings provide a deeper understanding of endogenous RNA transport in legume-rhizobia symbiotic processes.

关键词： symbiosis nodulation systemic transport of RNAs soybean rhizobia

A comprehensive overview of cotton genomics,biotechnology and molecular biological studies

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Science China(Life Sciences) 2023年第10期66卷 2214-2256页

作者： Xingpeng Wen Zhiwen Chen Zuoren Yang Maojun Wang Shuangxia Jin Guangda Wang Li Zhang Lingjian Wang Jianying Li Sumbul Saeed Shoupu He Zhi Wang Kun Wang zhaosheng kong Fuguang Li Xianlong Zhang Xiaoya Chen Yuxian Zhu Institute for Advanced Studies Wuhan UniversityWuhan 430072China College of Life Sciences Wuhan UniversityWuhan 430072China National Key Laboratory of Plant Molecular Genetics CAS Center for Excellence in Molecular Plant SciencesUniversity of CASChinese Academy of SciencesShanghai 200032China Hainan Yazhou Bay Seed Laboratory Sanya 572025China State Key Laboratory of Cotton Biology Institute of Cotton ResearchChinese Academy of Agricultural SciencesAnyang 455000China Hubei Hongshan Laboratory National Key Laboratory of Crop Genetic ImprovementCollege of Plant Science and TechnologyHuazhong Agricultural UniversityWuhan 430070China State Key Laboratory of Plant Genomics Institute of MicrobiologyInnovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China Shanxi Agricultural University Jinzhong 030801China

Cotton is an irreplaceable economic crop currently domesticated in the human world for its extremely elongated fiber cells specialized in seed epidermis,which makes it of high research and application value.To date,numerous research on cotton has navigated various aspects,from multi-genome assembly,genome editing,mechanism of fiber development,metabolite biosynthesis,and analysis to genetic breeding.Genomic and 3D genomic studies reveal the origin of cotton species and the spatiotemporal asymmetric chromatin structure in fibers.Mature multiple genome editing systems,such as CRISPR/Cas9,Cas12(Cpf1)and cytidine base editing(CBE),have been widely used in the study of candidate genes affecting fiber development.Based on this,the cotton fiber cell development network has been preliminarily drawn.Among them,the MYB-b HLH-WDR(MBW)transcription factor complex and IAA and BR signaling pathway regulate the initiation;various plant hormones,including ethylene,mediated regulatory network and membrane protein overlap fine-regulate elongation.Multistage transcription factors targeting Ces A 4,7,and 8 specifically dominate the whole process of secondary cell wall thickening.And fluorescently labeled cytoskeletal proteins can observe real-time dynamic changes in fiber development.Furthermore,research on the synthesis of cotton secondary metabolite gossypol,resistance to diseases and insect pests,plant architecture regulation,and seed oil utilization are all conducive to finding more high-quality breeding-related genes and subsequently facilitating the cultivation of better cotton varieties.This review summarizes the paramount research achievements in cotton molecular biology over the last few decades from the above aspects,thereby enabling us to conduct a status review on the current studies of cotton and provide strong theoretical support for the future direction.

关键词： cotton genome assembly genome editing fiber development gossypol plant architecture oil utilization genetic breeding

Towards a better recording of microtubule cytoskeletal spatial organization and dynamics in plant cells

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Journal of Integrative Plant Biology 2019年第4期61卷 388-393页

作者： Weiwei Liu Chaofeng Wang Guangda Wang Yinping Ma Juan Tian Yanjun Yu Li Dong zhaosheng kong State Key Laboratory of Plant Genomics Institute of Microbiology the Chinese Academy of Sciences The University of Chinese Academy of Sciences

Numerous fluorescent marker lines are currently available to visualize microtubule(MT)architecture and dynamics in living plant cells, such as markers expressing p35S::GFP-MBD or p35S::GFP-TUB6.However, these MT marker lines display obvious defects that affect plant growth or produce unstable fluorescent signals. Here, a series of new marker lines were developed, including the pTUB6::VisGreen-TUB6-expressing line in which TUB6 is under the control of its endogenous regulatory elements and e GFP is replaced with VisGreen, a brighter fluorescent protein. Moreover, two different markers were combined into one expression vector and developed two dual-marker lines.These marker lines produce bright, stable fluorescent signals in various tissues, and greatly shorten the screening process for generating dual-marker lines.These new marker lines provide a novel resource for MT research.

关键词： microtubule cytoskeletal plant cells^FA

GhAlaRP, a cotton alanine rich protein gene, involves in fiber elongation process

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The Crop Journal 2021年第2期9卷 313-324页

作者： Shouhong Zhu Yanjun Li Xinyu Zhang Feng Liu Fei Xue Yongshan Zhang zhaosheng kong Qian-Hao Zhu Jie Sun College of Agriculture/The Key Laboratory of Oasis Eco-agriculture Shihezi UniversityShihezi 832000XinjiangChina State Key Laboratory of Cotton Biology Institute of Cotton ResearchAcademy of Agricultural SciencesAnyang 455000HenanChina State Key Laboratory of Plant Genomics Institute of Microbiology Academy of Seed DesignChinese Academy of SciencesBeijing 100101China CSIRO Agriculture and Food GPO Box 1700Canberra 2601Australia

Fiber length is one of the most important quality parameters of cotton fibers.Transcriptomic analyses of developing cotton fibers have identified genes preferentially expressing in fiber elongation stage,but few have been functionally characterized.Here,on the basis of confirmation of the preferential expression profile of GhAlaRP(Gh_A09G1166 and Gh_D09G1172),an alanine rich protein gene,in the rapid elongating fibers,we investigated the role of GhAlaRP in fiber development by generating transgenic cottons with an increased or decreased expression level of GhAlaRP.Our results showed that the fiber length was consistently significantly shorter in both the GhAlaRP-RNAi lines and the alarp mutant generated by genome editing than in the control YZ-1.GhAlaRP was localized on plasma membrane,nucleus and endoplasmic reticulum.The yeast two-hybrid assay and bimolecular fluorescence complementation assay showed that GhAlaRP co-expresses and interacts with GhAnnexin(Gh_D11G2184)and GhEXPA(Gh_A10G2323)that are involved in fiber elongation.Down-regulation of GhAlaRP co-suppressed the expression levels of GhAnnexin and GhEXPA.These results suggest a role of GhAlaRP in regulation of cotton fiber elongation,which could be achieved by regulating the functions of GhAnnexin and GhEXPA.

关键词： AlaRP Fiber elongation G.hirsutum Genome editing

Kinesin-4 Functions in Vesicular Transport on Cortical Microtubules and Regulates Cell Wall Mechanics during Cell Elongation in Plants

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Molecular Plant 2015年第7期8卷 1011-1023页

作者： zhaosheng kong Motohide Ioki Siobhan Braybrook Shundai Li Zheng-Hua Ye Yuh-Ru Julie Lee Takashi Hotta Anny Chang Juan Tian Guangda Wang Bo Liu Department of Plant Biology University of California Davis CA 95616 USA State Key Laboratory of Plant Genomics Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China Sainsbury Laboratory Cambridge University of Cambridge Cambridge CB2 1LR UK Department of Biochemistry and Molecular Biology Pennsylvania State University State College PA 16802 USA Department of Plant Biology University of Georgia Athens GA 30602 USA Present address: Center for Environmental Biology and Ecosystem Studies National Institute for Environmental Studies Onogawa 16-2 Tsukuba Ibaraki305-8506 Japan Present address: Nara Institute of Science and Technology Graduate School of Biological Sciences 8916-5 Takayama Ikoma Nara 630-0192 Japan

In plants, anisotropic cell expansion depends on cortical microtubules that serve as tracks along which macromolecules and vesicles are transported by the motor kinesins of unknown identities. We used cotton （Gossypium hirsutum） fibers that underwent robust elongation to discover kinesins that are involved in cell elongation and found Gh KINESIN-4A expressed abundantly. The motor was detected by immunofluores- cence on vesicle-like structures that were associated with cortical microtubules. In Arabidopsis thaliana, the orthologous motor At KINESIN-4A/FRA1, previously implicated in cellulose deposition during second- ary growth in fiber cells, was examined by live-cell imaging in cells expressing the fluorescently tagged functional protein. The motor decorated vesicle-like particles that exhibit a linear movement along cortical microtubules with an average velocity of 0.89 l^m/min, which was significantly different from those linked to cellulose biosynthesis. We also discovered that At KINESIN-4A/FRA1 and the related At KINESIN-4C play redundant roles in cell wall mechanics, cell elongation, and the axial growth of various vegetative and reproductive organs, as the loss of At KINESIN-4C greatly enhanced the defects caused by a null mutation at the KINESIN-4A/FRA 1 locus. The double mutant displayed a lack of cell wall softening at normal stages of rapid cell elongation. Furthermore, enhanced deposition of arabinose-containing carbohydrate was detected in the kinesin-4 mutants. Our findings established a connection between the Kinesin-4-based transport of cargoes containing non-cellulosic components along cortical microtubules and cell wall mechanics and cell elongation in flowering plants.

关键词： cytoskeleton cytoskeleton-cell wall interactions leaf/vegetative development Arabidopsis cotton

Identification, Expression and Functional Analysis of a Receptor-like Cytoplasmic Kinase, OsRLCK1, in Rice

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Journal of Integrative Plant Biology 2007年第6期49卷 898-907页

作者： zhaosheng kong Wenying Xu Qun Li Yongbiao Xue Laboratory of Molecular and Developmental Biology Institute of Genetics and Developmental Biology the Chinese Academy of Sciences Beijing 100101 China

Pollination involves a series of complex cellular interactions and signal transduction events. Numerous reports have suggested a central role for protein kinases in pollen germination and pollen tube growth and a large number of receptor-like kinases have been detected exclusively in pollen in higher plants. However, few are well characterized, especially for the receptor-like cytoplasmic kinases. Here we report a receptor-like kinase gene, OsRLCK1, which belongs to the receptor-like cytoplasmic kinase Ⅷ subfamily. Real-time quantitative polymerase chain reaction analysis and whole mount RNA in situ hybridization showed that OsRLCK1 is a pollen-specific gene and expressed only in the mature pollen. When expressed in the onion epidermal cells, the OsRLCK1-GFP fusion protein was diffused throughout the cell, indicating its cytoplasmic and nuclear localization. The Maltose Binding Protein-OsRLCK1 recombinant protein was found to be capable of autophosphorylation on threonine residue, showing that it encodes a functional kinase. These results suggest that OsRLCK1 is likely to play a role in a signaling pathway associated with pollen performance during pollination in rice.

关键词： pollination receptor-like cytoplasmic kinase rice （Oryza sativa）.

Cytoskeletal control of trichome cell shaping: from Arabidopsis trichomes to cotton fibers

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Cytoskeletal control of trichome cell shaping: from Arabidop...