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Journal of Integrative plant Biology 2010年第10期52卷 933-940页

作者： Neila Jellouli Asma Ben Salem Abdelwahed Ghorbel Hatem Ben Jouira Laboratoire de Physiologie Moléculaire de la Vigne Centre de Biotechnologie de Borj Cédria

An efficient protein extraction method is crucial to ensure successful separation by two-dimensional electrophoresis （2-DE） for recalcitrant plant species, in particular for grapevine （Vitis vinifera L.）. Trichloroacetic acid-acetone （TCA-acetone） and phenol extraction methods were evaluated for proteome analysis of leaves and roots from the Tunisian cultivar ＇Razegui＇. The phenol-based protocol proved to give a higher protein yield, a greater spot resolution, and a minimal streaking on 2-DE gels for both leaf and root tissues compared with the TCA-based protocol. Furthermore, the highest numbers of detected proteins on 2-DE gels were observed using the phenol extraction from leaves and roots as compared with TCA-acetone extraction.

关键词： plant proteins VITIS vinifera LEAVES ROOTS (Botany) plant extracts ELECTROPHORESIS SEPARATION (Technology) PHENOLS EXTRACTION (Chemistry) GRAPES plant proteomics

A Point Mutation in the Pentatricopeptide repeat Motif of the AtECB2 Protein Causes Delayed Chloroplast Development

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Journal of Integrative plant Biology 2011年第4期53卷 258-269页

作者： Zhi-Lin Cao Qing-Bo Yu Yue Sun Yang Lu Yong-Lan Cui Zhong-Nan Yang College of Life and Environmental Sciences Shanghai Normal University Shanghai 200234 China Department of Resource and Environmental Engineering Henan Institute of Engineering Zhengzhou 451191 China

AtECB2 encodes a pentatricopeptide repeat（PPR） protein that regulates the editing of the plastid genes accD and *** ecb2-1 knockout shows an albino phenotype and is seedling *** this study, we isolated an allelic mutant of the AtECB2 gene,ecb2-2,which showed delayed greening phenotype but could complete their life *** this mutant,the Thr^500 is converted to Ile^500 in the 13^th PPR motif of the AtECB2 *** electron microscopy demonstrated that chloroplast development was delayed in both the cotyledons and leaves of the *** investigation of the chloroplast gene expression profile indicated that PEP（plastid-encoded RNA polymerase） activity in ecb2-2 cotyledons was not obviously affected,whereas it was severely impaired in *** result suggests that the PEP activities cause the different phenotypes of the ecb2-1 and ecb2-2 *** editing efficiency of the three editing sites of accD（C794 and C1568） and ndhF（C290） in the mutant was dynamically altered, which was in agreement with the *** result indicates that the editing efficiency of accD and ndhF in the ecb2-2 mutant is associated with a delayed greening *** ecb2-2 can survive and set seeds,this mutant can be used for further investigation of RNA editing and chloroplast development in arabidopsis.

关键词： PENTATRICOPEPTIDE repeat genes CHLOROPLASTS plant proteins MUTATION (Biology) RNA editing ARABIDOPSIS TRANSMISSION electron microscopy PLACENTA

Proteomic Study Identifies proteins Involved in Brassinosteroid Regulation of Rice Growth

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Journal of Integrative plant Biology 2010年第12期52卷 1075-1085页

作者： Fengru Wang Ming-Yi Bai Zhiping Deng Juan A. Oses-Prieto Alma L. Burlingame Tiegang Lu Kang Chong Zhi-Yong Wang Key Laboatory of PhotoSynthesis and Environmental Molecular Physiology Institute of Botany the Chinese Academy of Sciences Beijing 100093 China Agriculture University of Hebei Baoding 071001 China Department of plant Biology Carnegie Institution for Science Stanford CA 94305 USA Mass Spectrometry Facilityl Department of Pharmaceutical Chemistry University of California San Francisco CA 94143 USA Biotechnology Research Institute the Chinese Academy of Agriculture Sciences Beijing 100081 China Graduate University of the Chinese Academy of Sciences Beijing 100049 China

Brassinosteroids （BRs） are essential hormones for growth and development of plant. In rice, BRs regulate multiple developmental processes and affect many important traits such as height, leaf angle, fertility and seed filling. We identified brassinosteroid-regulated proteins in rice using proteomic approaches and performed functional analysis of some BR-regulated proteins by overexpression experiments. Using two-dimensional difference gel electrophoresis （2-D DIGE） followed by protein identification by mass spectrometry, we compared proteomic differences in the shoots and roots of the BR-insensitive mutant d61-4 and BR-deficient mutant brd1-3. We identified a large number of proteins differentially expressed in the mutants compared with wild type control. These include a glycine-rich RNA-binding protein （OsGRP1） and a DREPP2 protein, which showed reduced levels in the BR mutants. Overexpression of these two proteins partially suppressed the dwarf phenotype of the Arabidopsis BR-insensitive mutant bri1-5. In contrast to the reduced protein level, the RNA level of OsGRP1 was not significantly affected in the BR mutants or by BR treatment, suggesting BR regulation of OsGRP1 at the posttranslational level. This study identifies many BR-regulated proteins and demonstrates that OsGRP1 functions downstream in the BR signal transduction pathway to promote cell expansion.

关键词： PROTEOMICS plant proteins BRASSINOSTEROIDS plant growth -- Regulation RICE -- Varieties TWO-dimensional electrophoresis MASS spectrometry

Defining the Mitochondrial Stress Response in Arabidopsis thaliana

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Molecular plant 2009年第6期2卷 1310-1324页

作者： Olivier Van Aken Botao Zhang Chris Carrie Vindya Uggalla Ellen Paynter Estelle Giraud James Whelan ARC Centre of Excellence in plant Energy Biology MCS Building M316 University of Western Australia 35 Stirling Highway Crawley 6009 Western AustraliaAustralia

To obtain a global overview of how mitochondria respond to stress, we aimed to define the plant mitochondrial stress response （MSR）. By combining a set of 1196 Arabidopsis thaliana genes that putatively encode mitochondrial proteins with 16 microarray experiments on stress-related conditions, 45 nuclear encoded genes were defined as widely stress-responsive. Using green fluorescent protein （GFP） fusion assays, the mitochondrial targeting of a large number of these proteins was tested, confirming in total 26 proteins as mitochondrially targeted. Several of these proteins were observed to be dual targeted to mitochondria and plastids, including the small heat shock proteins sHSP23.5 and sHSP23.6. In addition to the well defined stress components of mitochondria, such as alternative oxidases, nicotinamide adenine dinudeotide （NAD（P）H） dehydrogenases, and heat shock proteins, a variety of other proteins, many with unknown function, were identified. The mitochondrial carrier protein family was over-represented in the stress-responsive genes, suggesting that stress induces altered needs for metabolite transport across the mitochondrial inner membrane. Although the genes encoding many of these proteins contain common cis-acting regulatory elements, it was apparent that a number of distinct regulatory processes or signals likely triggered the MSR. Therefore, these genes provide new model systems to study mitochondrial retrograde regulation, in addition to the widely used alternative oxidase model. Additionally, as changes in proteins responsive to stress did not correlate well with changes at a transcript level, it suggests that post-transcriptional mechanisms also play an important role in defining the MSR.

关键词： enzymes genes green fluorescent protein heat shock proteins mitochondria oxidoreductases plant proteins plastids stress stress response Arabidopsis thaliana Genes Heat Shock proteins Plastid stress response Arabidopsis thaliana Oxidoreductases plant proteins proteins Enzymes Mitochondria Green Fluorescent proteins

LEUNIG_HOMOLOG and LEUNIG Regulate Seed Mucilage Extrusion in Arabidopsis

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Journal of Integrative plant Biology 2011年第5期53卷 399-408页

作者： Minh Bui Nathan Lim Paja Sijacic Department of Cell Biology and Molecular Genetics University of MarylandCollege ParkMaryland 20742USA

LEUNIG （LUG） and LEUNIG_HOMOLOG （LUH） encode two closely related Arabidopsis proteins, belonging to the Gro/TLE family of transcriptional co-repressors. These two genes were previously shown to exhibit partially overlapping functions in embryo and flower development. In this report, the role of both LUH and LUG on seed mucilage extrusion was examined. Seed mucilage extrusion occurs after the seeds are imbibed, serving as functional aid in seed hydration, germination, and dispersal. While luh-1 mutants exhibited strong defects in seed mucilage extrusion, lug-3 mutants exhibited a minor phenotype in mucilage extrusion. Further characterization indicates that luh-1 does not exhibit any obvious defect in seed epidermal cell differentiation, mucilage synthesis, or mucilage deposition, suggesting a specific role of LUH in mucilage extrusion. This seed mucilage phenotype of luh-1 is identical to that of mucilage modified 2 （mum2） mutants. MUM2 encodes a β-galactosidase required for the modification of the mucilage. Quantitative reverse transcription polymerase chain reaction of RNA extracted from siliques detected a slight decrease of MUM2 mRNA in the luh-1 mutant compared to the wild type. Together, LUH and possibly LUG may specifically regulate mucilage extrusion by promoting the expression of genes required for mucilage maturation.

关键词： ARABIDOPSIS plant proteins GENETIC transcription plant genetics GENETIC repressors plant embryology MUCILAGE plant mutation

WBC27,an Adenosine Tri-phosphate-binding Cassette Protein,Controls Pollen Wall Formation and Patterning in Arabidopsis

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Journal of Integrative plant Biology 2011年第1期53卷 74-88页

作者： Xiao-Ying Dou Ke-Zhen Yang Yi Zhang Wei Wang Xiao-Lei Liu Li-Qun Chen Xue-Qin Zhang De Ye State Key Laboratory of plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China Key Laboratory of Biotechnology and Crop Quality Improvement Ministry of Agriculture Southwest University Chongqing 400716 China

In flowering plants, the exine components are derived from tapetum. Despite its importance to sexual plant reproduction, little is known about the translocation of exine materials from tapetum to developing microspores. Here we report functional characterization of the arabidopsis WBC27 gene. WBC27 encodes an adenosine tri-phosphate binding cassette （ABC） transporter and is expressed preferentially in tapetum. Mutation of WBC27 disrupted the exine formation. The wbc27 mutant microspores began to degenerate once released from tetrads and most of the microspores collapsed at the uninucleate stage. Only a small number of wbc27-1 microspores could develop into tricellular pollen grains. These survival pollen grains lacked exine and germinated in the anther before anthesis. All of these results suggest that the ABC transporter, WBC27 plays important roles in the formation of arabidopsis exine, possibly by translocation of lipidic precursors of sporopollenin from tapetum to developing microspores.

关键词： ADENOSINE triphosphate ARABIDOPSIS plant proteins EXINE plant reproduction plant gene expression plant mutation

At Long Last： Evidence for Pexophagy in plants

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Molecular plant 2014年第8期7卷 1257-1260页

作者： Tamar Avin-Wittenberg Alisdair R. Fernie Max-Planck-Institute of Molecular plant Physiology Am Mühlenberg 1 14476 Potsdam-Golm Germany

Peroxisomes are highly dynamic single-membrane-bound eukaryotic organelles displaying great variability in enzymatic content （Platta and Erdmann, 2007）. plant peroxisomes function in a plethora of crucial development and stress ameliorating processes. Among the functions of plant peroxisomes are 13-oxidation of fatty acids, phytohormone production, participation in photorespiration, the glyoxyalte cycle, detoxification processes, and signal molecule generation （Hu et al., 2012）.

关键词： plant cell microbodies AUTOPHAGIC vacuoles plant proteins plant organelles plant vacuoles SEEDS -- Growth

Visualization of MicrotubuleOrganization and Dynamics in Living Arabidopsis Embryonic Cells

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Molecular plant 2014年第8期7卷 1397-1401页

作者： Qiannan Wang Shanjin Huang Key Laboratory of plant Molecular Physiology Institute of Botany Chinese Academy of Sciences Beijing 100093 China University of Chinese Academy of Sciences Beijing 100049 China National Center for plant Gene Research Beijing 100101 China

Dear Editor, Embryogenesis is a critical developmental stage during the life cycle of flowering plants. During embryogenesis, the first round of asymmetric cell division in the zygote is followed by a series of cellular events, including cell division （symmetric or asymmetric） and directional cell expan- sion to generate the apical-basal axis, radial and lateral symmetry, and patterns of different cell fates for initiation of different organ primordia, which lay the foundation for post-embryonic development （Jurgens, 2001; Wendrich and Weijers, 2013）.

关键词： plant microtubules plant embryology ARABIDOPSIS thaliana EMBRYONIC stem cells plant proteins MEMBRANE fusion plant gene expression

Structural Basis of Interaction of Bipartite Nuclear Localization Signal from Agrobacterium VirD2 with Rice Importin-o

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Molecular plant 2014年第6期7卷 1061-1064页

作者： Chiung-Wen Chang, Simon J. Williams, Rafael Miguez Cou?ago Bostjan Kobe School of Chemistry and Molecular Biosciences Institute for Molecular Bioscience and Australian Infectious Diseases ResearchCentre University of Queensland Brisbane Qld 4072 Australia

Dear Editor, In the eukaryotic cell, the nuclear envelope separates genomic DNA from the rest of the cell, but ensures a molecu- lar communication between the nucleus and the cytoplasm through the nuclear pore complexes （NPCs）. Nuclear trans- port of macromolecules is an energy-dependent process, exemplified by the classical nuclear import pathway. In the cytoplasm, proteins containing classical nuclear localization signals （cNLSs） are recognized by a hetero-dimeric transport receptor （importin-α （Impα）：importin-β （Impl3） complex）. Impa is employed as an adaptor protein that specifically rec-ognizes the classical nuclear localization signal （cNLS） from the cargo protein （Marfori et al., 2011）. Translocation of the trimeric cargo：transport receptor import complex through the NPCs is mediated by transient interactions between Impβ and nucleoporins in the NPC. The directionality of the trans- port is imparted by the asymmetric distribution, between the cytoplasm and the nucleus, of the distinct nucleotide-bound （GTP/GDP） states of the small GTPase Ran （Tran et al., 2007）.

关键词： AGROBACTERIUM RESEARCH plant proteins plant proteins -- Structure plant cell nuclei CRYSTAL structure CARRIER proteins -- Molecular structure CARRIER proteins RICE -- Research

OsRFPH2-10,a RING-H2 Finger E3 Ubiquitin Ligase,Is Involved in Rice Antiviral Defense in the Early Stages of Rice dwarf virus Infection

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Molecular plant 2014年第6期7卷 1057-1060页

作者： Lifang Liu Lian Jin Xiahe Huang Yongtao Geng Feng Li Qingqing Qin Rui Wang Shaoyi Ji Shanshan Zhao Qi Xie Chunhong Wei Can Xie Biao Ding Yi Li The State Key Laboratory of Protein and plant Gene Research College of Life SciencesPeking UniversityBeijing 100871People’s Republic of China Peking-Yale Joint Center for plant Molecular Genetics and Agrobiotechnology College of Life SciencesPeking UniversityBeijing 100871People’s Republic of China Institute of Genetics and Developmental Biology Chinese Academy of SciencesBeijing 100101People’s Republic of China Department of Molecular Genetics and Center for RNA Biology The Ohio State UniversityColumbusOH 43210USA

Dear Editor,Rice dwarf virus(RDV),a member of the Phytoreoviurs genus,is transmitted to rice(Oryza sativa)plants by leaf-hopper(Nephotettix cincticeps)in a propagative *** by RDV results in severe stunting growth pheno-types and a dramatic reduction in grain *** genome of RDV is composed of 12-segmented double-stranded RNAs(S1-S12 based on their migration rates in agarose gel electro-phoresis).The 12 segments encode seven structural proteins(P1,P2,P3,P5,P7,P8,and P9 as the products of Sl,S2,S3,S5,S7,S8,and S9,respectively)and five nonstructural proteins(Pns4,Pns6,Pnsl0,Pns11,and Pns12 as the products of S4,S6,S10,S11,and S12,respectively).The outer capsid protein P2 is essential for RDV infection of insects and thus influences transmission of RDV by the insect vector(Omura et al.,1998;Zhou et al.,2007).P2 also contributes to the dwarf phenotype of infected rice by interfering with gibberellic acid synthesis(Zhu et al.,2005).When RDV-infected rice plants were main-tained via vegetative propagation for several years without insect transmission,they regained normal growth height due to loss of RDV P2 and Pnsl0 proteins(Pu et al.,2011).

关键词： RICE -- Disease & pest resistance RESEARCH RICE -- Diseases & pests UBIQUITIN ligases plant viruses plant proteins plant cells & tissues PHENOTYPE ANTIVIRAL agents