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Chinese Science Bulletin 2013年第23期58卷 2855-2861页

作者： MA LiJuan yang zhongnan ZHANG Sen College of Life and Environmental Sciences Shanghai Normal University

Microsporogenesis in flowering plants plays important roles in sexual reproduction. It has been reported that DEFECTIVE IN EXINE FORMATION1 (DEX1) is essential for exine pattern formation in Arabidopsis thaliana. However, the functions of DEX1 in regulating microspore development are largely not understood. In this study, we show that DEX1 is strongly expressed in the tapetum by using RNA in situ hybridization. dex1 microspores were degenerated and aborted after release from the tetrads. The callose wall in tetrads was thinner in the dex1 mutant than in the wild type, suggesting that DEX1 affects callose formation at the tetrad stage during anther development. RT-PCR and real-time PCR analyses showed that CalS5, which plays an important role in callose synthesis during microspore development, was greatly down-regulated in dex1 plants. DEX1 encodes a membrane protein with one transmembrane domain, one intracellular domain and one extracellular domain. Collectively, our results demonstrate that DEX1 is essential for microspore development, possibly by regulating the expression of CalS5.

关键词：小孢子发育拟南芥 Arabidopsis 蛋白质本地化 RNA原位杂交质膜 RT-PCR

A Single Amino-Acid Substitution at Lysine 40 of an Arabidopsis thaliana α-tubulin Causes Extensive Cell Proliferation and Expansion Defects

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Journal of Integrative Plant Biology 2013年第3期55卷 209-220页

作者： Xue Xiong Deyang Xu zhongnan yang Hai Huang Xiaofeng Cui National Laboratory of Plant Molecular Genetics Shanghai Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of Sciences College of Life and Environmental Sciences Shanghai Normal University

Microtubules are highly dynamic cytoskeletal polymers of α/β-tubulin heterodimers that undergo multiple post-translational modifications essential for various cellular functions in eukaryotes. The lysine 40 （K40） is largely conserved in α-tubulins in many eukaryote species, and the post-translational modification by acetylation at K40 is critical for neuronal development in vertebrates. However, the biological function of K40 of α-tubulins in plants remains unexplored. In this study, we show in Arabidopsis thaliana that constitutive expression of mutated forms of α-tubulin6 （TUA6） at K40 （TUA6K40A or TUA6K40Q ）, in which K40 is replaced by alanine or glutamine, result in severely reduced plant size. Phenotypic characterization of the 35S：TUA6K40A transgenic plants revealed that both cell proliferation and cell expansion were affected. Cytological and biochemical analyses showed that the accumulation of α- and β-tubulin proteins was significantly reduced in the transgenic plants, and the cortical microtubule arrays were severely disrupted, indicating that K40 of the plant α-tubulin is critical in maintaining microtubule stability. We also constructed 35S：TUA6K40R transgenic plants in which K40 of the engineered TUA6 protein is replaced by an arginine, and found that the 35S：TUA6K40R plants were phenotypically indistinguishable from the wild-type. Since lysine and arginine are similar in biochemical nature but arginine cannot be acetylated, these results suggest a structural importance for K40 of α-tubulins in cell division and expansion.

关键词： Arabidopsis α-tubulin expansion cell proliferation microtubule,

ATPG is required for the accumulation and function of chloroplast ATP synthase in Arabidopsis

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Chinese Science Bulletin 2013年第26期58卷 3224-3232页

作者： KONG MengMeng WANG FenFei yang zhongnan MI HuaLing National Key Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences College of Life and Environmental Sciences Shanghai Normal University

The subunit Ⅱ of chloroplast ATP synthase is one of the two peripheral stalks, which associates the catalytic CF1 with mem-brane-spanning CFo . Although the structural and functional roles of chloroplast ATP synthase have been extensively examined, the physiological significance of subunit Ⅱ in vivo is still unclear. In this work, we identified one Arabidopsis T-DNA insertion mutant of atpG gene encoding the subunit Ⅱ of chloroplast ATP synthase. The atpg null mutant displayed an albino lethal pheno-type, as it could not grow photoautotrophically. Transmission electron microscopy analysis showed that chloroplasts of atpg lacked the organized thylakoid membranes. Loss of subunit Ⅱ affected the accumulation of CF1-CFo complex, however, it did not seem to have an effect on the CF1 assembly. The light induced ATP formation of atpg was significantly reduced compared with the wild type. Based on these results, we suggested that ATPG was essential for the accumulation and function of chloroplast ATP synthase.

关键词： ATP合酶 ATPG 叶绿体拟南芥积累透射电子显微镜分析 T-DNA插入类囊体膜

Prediction of anther-expressed gene regulation in Arabidopsis

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Chinese Science Bulletin 2008年第20期53卷 3198-3203页

作者： HUANG JiFeng yang JingJin WANG Guan YU QingBo yang zhongnan Mathematics & Science College Shanghai Normal University Shanghai 200234 China College of Life and Environmental Sciences Shanghai Normal University ShangHai 200234 China

Anther development in Arabidopsis, a popular model plant for plant biology and genetics, is controlled by a complex gene network. Despite the extensive use of this genus for genetic research, little is known about its regulatory network. In this paper, the direct transcriptional regulatory relationships between genes expressed in Arabidopsis anther development were predicted with an integrated bioinformatic method that combines mining of microarray data with promoter analysis. A total of 7710 transcription factor-gene pairs were obtained. The 80 direct regulatory relationships demonstrating the highest confidence were screened from the initial 7710 pairs; three of the 80 were validated by previous experiments. The results indicate that our predicted results were reliable. The regulatory relationships revealed by this research and described in this paper may facilitate further investigation of the molecular mechanisms of anther development. The bioinformatic method used in this work can also be applied to the prediction of gene regulatory relationships in other organisms.

关键词：植物生长发育花粉囊基因调节生物信息学

OsMYB103 is required for rice anther development by regulating tapetum development and exine formation

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Chinese Science Bulletin 2010年第29期55卷 3288-3297页

作者： ZHANG Sen FANG ZiJun ZHU Jun GAO JuFang yang zhongnan College of Life and Environment Sciences Shanghai Normal University Shanghai 200234 China

The Arabidopsis AtMYB103 gene is required for anther development,but whether the homologous gene in rice has the same role is *** analysis indicated that the rice OsMYB103 gene shares a high sequence similarity with ***,we investigated the functional role of OsMYB103 in anther development using an RNAi *** OsMYB103 RNA transcript was expressed most abundantly in flowers,specifically in the tapetum,premeiotic pollen mother cells,and meiotic ***103-RNAi transgenic lines grew normally during their vegetative phase but displayed reduced male fertility,a phenotype that was associated with downregulated OsMYB103 transcript *** of OsMS2,an ortholog of the Arabidopsis AtMS2 gene,was also dramatically reduced in the transgenic *** of OsMYB103 led to defects in tapetum development,and most of the microspores in mature anthers lacked ***,OsMYB103 could partially rescue the male sterility phenotype of an AtMYB103 knockout mutant *** together,these results indicate that OsMYB103 does have an important role in rice tapetum and microspore development.

关键词：花药绒毡层水稻 RNA干扰转基因植株同源基因外壁转录水平序列相似性

Polymerase chain reaction of Au nanoparticle-bound primers

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Chinese Science Bulletin 2005年第18期50卷 2016-2020页

作者： SHEN Hebai HU Min yang zhongnan WANG Chen ZHU Longzhang Life and Environment Science College Shanghai Normal University Shanghai 200234 China

Polymerase chain reaction (PCR) is a useful technique for in vitro amplification of a DNA fragment. In this paper, a PCR procedure using Au nanoparticle (AuNP) -bound primers was systemically studied. The 5′-SH- (CH2)6-modified primers were covalently attached to the AuNP surface via Au-S bonds, and plasmid pBluescript SK was used as a template. The effects of the concentration of AuNP-bound primers, annealing temperature and PCR cy-cles were evaluated, respectively. The results indicate that PCR can proceed successfully under optimized condition, with either forward or reverse primers bound to the AuNP surface or with both the two primers bound to the AuNP surface. Development of PCR procedure based on AuNPs not only makes the isolation of PCR products very convenient, but also provides novel methods to prepare AuNP-bound ssDNA and nanostructured material.

关键词：聚合酶链反应 PCR 金纳米离子退火温度优化条件

The Arabidopsis pentatricopeptide repeat protein PDM1 is associated with the intergenic sequence of S11-rpoA for rpoA monocistronic RNA cleavage

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Chinese Science Bulletin 2012年第26期57卷 3452-3459页

作者： YIN QianQian CUI YongLan ZHANG GuoRui ZHANG HongDao WANG XiaoMeng yang zhongnan College qf Life and Environmental Sciences Shanghai Normal UniversityShanghai 200234China

The PDM1 gene encodes a pentatricopeptide repeat protein of the PLS *** is essential for rpoA polycistronic processing in *** this study,we performed functional analysis of PDM1 in rpoA monocistronic cleavage and chloroplast *** pdm1 mutants display an albino lethal phenotype with severe chloroplast development *** the construct of PDM1 fused with the GFP gene was introduced into Arabidopsis protoplasts,the GFP signal was exclusively observed in *** shows that PDM1 is localized to *** the wild type,the rpoA transcript of about 990 nt is *** pdm1,this transcript is ***,Western blot showed that the RpoA protein in pdm1 is accumulated at a level approximately 1/3 of the wild *** suggests that certain other transcripts processed from L23-L2-S19-L22-S3-L16-L14S8-L36-S11-rpoA polycistronic precursor may be used as templates for protein *** determine whether PDM1 can bind to the rpoA pre-mRNA,we generated a transgenic Arabidopsis line with PDM1 fused to FLAG tag that was capable of complementing the pdm1-1 mutant *** immunoprecipitation analysis showed that PDM1 is associated with S11-rpoA intergenic *** indicates that PDM1 may bind to the S11-rpoA intergenic region to perform rpoA processing.

关键词：前体mRNA 转基因拟南芥多顺反子间隔序列蛋白质裂解 Western印迹叶绿体发育

Fine mapping of an Arabidopsis thaliana male sterile mutant EC2-157

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Frontiers in Biology 2006年第3期1卷 270-274页

作者： LIU Huijuan ZHANG Zaibao LI Hui GAO Jufang yang zhongnan College of Life and Environment Sciences Shanghai Teachers UniversityShanghai 200234China

An Arabidopsis thaliana male sterile mutant EC2-157 has been isolated using an EMS mutagenesis *** analysis indicated that it was controlled by a single recessive gene called *** pollen grains have been observed in mutant ***157 Has been mapped to a region of 74 kb located in BAC clone T6K22 on chro-mosome IV using a map-based cloning *** no male sterile genes have been reported in this region,ms157 could be a novel gene related to *** further molecular cloning and functional analysis on this gene should facili-tate our understanding of *** anther development.

关键词： EC2-157 anther development male sterile map-based cloning Arabidopsis thaliana

SLIDE, the Protein Interacting Domain of Imitation Switch Remodelers, Binds DDT-Domain Proteins of Different Subfamilies in Chromatin Remodeling Complexes

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Journal of Integrative Plant Biology 2013年第10期55卷 928-937页

作者： Jiaqiang Dong Zheng Gao Shujing Liu Guang Li zhongnan yang Hai Huang Lin Xu National Laboratory of Plant Molecular Genetics Institute of Plant Physiology & Ecology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences College of Life and Environment Sciences Shanghai Normal University

The Imitation Switch （ISWI） type adenosine triphosphate （ATP）-dependent chromatin remodeling factors are conserved proteins in eukaryotes, and some of them are known to form stable remodeling complexes with members from a family of proteins, termed DDT-domain proteins. Although it is well documented that ISWIs play important roles in different biological processes in many eukaryotic species, the molecular basis for protein interactions in ISWI complexes has not been fully addressed. Here, we report the identification of interaction domains for both ISWI and DDT-domain proteins. By analyzing CHROMATIN REMODELING11 （CH R11） and RINGLET1 （RLT1）, an Arabidopsis thaliana ISWI （AtlSWI） and AtDDT-domain protein, respectively, we show that the SLIDE domain of CHR11 and the DDT domain together with an adjacent sequence of RLT1 are responsible for their binding. The Arabidopsis genome contains at least 12 genes that encode DDT-domain proteins, which could be grouped into five subfamilies based on the sequence similarity. The SLIDE domain of AtlSWI is able to bind members from different AtDDT subfamilies. Moreover, a human ISWI protein SNF2H is capable of binding AtDDT-domain proteins through its SLIDE domain, suggesting that binding to DDT-domain proteins is a conserved biochemical function for the SLIDE domain of ISWIs in eukaryotes.

关键词： Arabidopsis chromatin remodeling factors DDT-domain proteins Imitation Switch protein interaction.

TAA family contributes to auxin production during de novo regeneration of adventitious roots from Arabidopsis leaf explants

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Science Bulletin 2016年第22期61卷 1728-1731页

作者： Beibei Sun Lyuqin Chen Jingchun Liu Xuening Zhang zhongnan yang Wu Liu Lin Xu National Key Laboratory of Plant Molecular Genetics CAS Center for Excellence in Molecular Plant Sciences Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032China College of Life and Environment Sciences Shanghai Normal University Shanghai 200234 China University of Chinese Academy of Sciences 19A Yuquan RoadBeijing 100049 China Department of Plant and Microbial Biology Ztirich-Basel Plant Science Center University of Zurich 8008 Zurich Switzerland

Many differentiated plant organs have the ability to regenerate into a new plant after detachment via de novo organogenesis. During de novo root organogenesis from Arabidopsis thaliana leaf explants, wounding first induces endogenous auxin production in mesophyll cells. Auxin is then polar transported to, and accumulates in, regenerationcompetent cells near the wound to trigger the cell-fate transition. The TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS(TAA) family proteins and the YUCCA(YUC) family proteins catalyze two successive biochemical steps in auxin biogenesis, and YUCs have been shown to be involved in auxin production in mesophyll cells during de novo root organogenesis. In thisstudy, we show that the TAA family is also required for adventitious rooting. Inhibition of TAA blocked adventitious root formation from leaf explants. Intriguingly,whereas YUC1 and YUC4 have been shown to be highly induced by wounding, TAA genes retained consistent expression levels before and after leaf ***, we suggest that TAAs and YUCs are both required for auxin biogenesis in leaf explants, but they play different roles in regeneration. While YUC1 and YUC4 function in response to wounding to catalyze the ratelimiting step in auxin biosynthesis, TAAs probably serve as abiding and basal enzymes during de novo root organogenesis from leaf explants.

关键词： De novo root organogenesis TAA1TAR2 Plant regeneration Auxin biogenesis Arabidopsis