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A Viral Protein Suppresses siRNA-directed Interference in Tobacco Mosaic Virus Infection

Acta Biochimica et Biophysica Sinica 2005年第4期37卷 248-253页

作者： Ming-Min ZHAO De-Rong AN Guang-Hua HUANG~1 Zu-Hua HE~2 Jiang-Ye CHEN~1 College of plant Protection,North West Science and Technology University of Agriculture and Forestry,Shaanxi 712100,china ~1state key laboratory of molecular Biology,institute of Biochemistry and Cell Biology,shanghai institutes for biological sciences. chinese academy of sciences,shanghai 200031,china ~2state key laboratory of plant molecular genetics.institute of plant physiology and ecology,shanghai institutes for biological sciences.chinese academy of sciences,shanghai 200032.china College of plant Protection North West Science and Technology University of Agriculture and ForestryShaanxi 712100 China state key laboratory of molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China state key laboratory of plant molecular genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

plant viruses encode suppressors of post-transcriptional gene silencing (PTGS), an adaptivedefense response that limits virus replication and its spread in *** helper component proteinase (HC-Pro) of the potato virus A (PVA,genus Potyvirus) suppresses PTGS of silenced ***,the effectof HC-Pro on siRNA-directed interference in the tobacco mosaic virus (TMV) was examined by using atransient Agrobacterium tumefaciens-based delivery system in intact *** was shown that the interferenceeffect was completely blocked by co-infiltration with HC-Pro plus siRNA constructs in both systemic andhypersensitive *** the system host,all plants agro-infiltrated with HC-Pro plus siRNA constructsdisplayed the same symptoms as the negative ***,TMV RNA accumulation was found to beabundant in the upper leaves using reverse transcriptase-PCR (RT-PCR) and Northern blot *** thecontrary,plants agro-infiltrated with the siRNA construct alone were free of ***,ourstudy suggests that the transient expression of HC-Pro inhibited the siRNA-directed host defenses againstTMV infection.

关键词： siRNA HC-Pro tobacco mosaic virus suppressor

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Expressional Analysis of an EREBP Transcription Factor Gene OsEBP-89 in Rice

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 21-26页

作者： Hui SHEN and Zong-Yang WANG( The state key laboratory of plant molecular genetics, institute of plant physiology & ecology, shanghai institutes for biological sciences,the chinese academy of sciences, shanghai 200032, china ) The state key laboratory of plant molecular genetics Institute of Plant Physiology & Ecology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences Shanghai 200032 China

OsEBP-89 gene encodes an ethylene responsive element binding protein (EREBP) transcription factor from rice (Oryza sativa). Northern blot analysis revealed that OsEBP-89 was expressed in root, stem, seeds, flowers and leaves. Histochemical assay showed that β-glucuronidase (GUS) was expressed mainly in phloem of vascular tissues of the root and stem transition region (RST), basal part of sheath roots, stem node and basal part of adventitious roots, also in endosperm of seeds in transgenic rice harboring OsEBP-89/GUS construct (pNSG). A sequence region from -279 to -97 was found to play an important role for OsEBP-89 gene expression through promoter deletion assay. The possible function of OsEBP-89 gene was discussed.

关键词： OsEBP-89 gene rice EREBP subfamily transcription factor tissue expression

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Transcriptional, post-transcriptional and post-translational regulations of gene expression during leaf polarity formation

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Cell Research 2007年第6期17卷 512-519页

作者： Lin Xu Li YangHai Huang National laboratory of plant molecular genetics, shanghai institute of plant physiology and ecology, shanghai institutes for biological sciences, chinese academy of sciences, 300 Fenglin Road, shanghai 200032, china National laboratory of plant molecular genetics Shanghai Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Present address: IBMP CNRS Universite Louis Pasteur 67084 Strasbourg France Present address: Department of Biology University of Pennsylvania Philadelphia PA 19104 USA

Leaf morphogenesis requires the establishment of adaxial-abaxial polarity after primordium initiation from the shootapical meristem (SAM). Several families of transcription factors are known to play critical roles in promoting adaxial orabaxial leaf fate. Recently, post-transcriptional gene silencing pathways have been shown to regulate the establishmentof leaf polarity, providing novel and exciting insights into leaf development. For example, microRNAs (miR165/166)and a trans-acting siRNA (TAS3-derived tasiR-ARF) have been shown to repress the expression of several key transcrip-tion factor genes. In addition, yet another level of regulation, post-translational regulation, has been revealed recently bystudies on the role of the 26S proteasome in leaf polarity. Although our understanding regarding the molecular mecha-nisms underlying establishment of adaxial-abaxial polarity has greatly improved, there is still much that remains *** review aims to discuss recent progress, as well as the remaining questions, regarding the molecular mechanismsunderlying leaf polarity formation.

关键词： Arabidopsis leaf development polarity formation

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Transiently Expressed Short Hairpin RNA Targeting 126 kDa Protein of Tobacco Mosaic Virus Interferes with Virus Infection

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Acta Biochimica et Biophysica Sinica 2006年第1期38卷 22-28页

作者： Ming-Min ZHAO~(1,2),De-Rong AN~(1*),Jian ZHAO~2,Guang-Hua HUANG~3,Zu-Hua HE~4,and Jiang-Ye CHEN~3 ~1 College of plant Protection,Northwest Science and Technology University of Agriculture and Forestry,Yangling 712100,china ~2 College of Agriculture,Yangtze University,Jingzhou 434025,china ~3 state key laboratory of molecular Biology,institute of Biochemistry and Cell Biology,shanghai institutes for biological sciences, chinese academy of sciences,shanghai 200031,china ~4 state key laboratory of plant molecular genetics.institute of plant physiology and ecology,shanghai institutes for biological sciences, chinese academy of sciences,shanghai 200032.china College of plant Protection Northwest Science and Technology University of Agriculture and Forestry Yang ling 712100 China College of Agriculture Yangtze University Jingzhou 434025 China state key laboratory of molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China state key laboratory of plant molecular genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China

RNA-interference (RNAi) silences gene expression by'guiding mRNA degradation in asequence-specific *** interfering RNA (siRNA),an intermediate of the RNAi pathway,has beenshown to be very effective in inhibiting virus infection in mammalian cells and cultured plant ***,wereport that Agrobacterium tumefaciens-mediated transient expression of short hairpin RNA (shRNA) couldinhibit tobacco mosaic virus (TMV) RNA accumulation by targeting the gene encoding the replication-asso-ciated 126 kDa protein in intact plant *** results indicate that transiently expressed shRNA efficientlyinterfered with TMV *** interference observed is sequence-specific,and time-and *** expressed shRNA corresponding to the TMV 126 kDa protein gene did not inhibit cucumbermosaic virus (CMV),an unrelated *** order to interfere with TMV accumulation in tobaccoleaves,it is essential for the shRNA constructs to be infiltrated into the same leaves as TMV *** support the view that RNAi opens the door for novel therapeutic procedures against virus *** propose that a combination of the RNAi technique and Agrobacterium-mediated transient expressioncould be employed as a potent antiviral treatment in plants.

关键词： tobacco mosaic virus 126 kDa protein RNA interference short hairpin RNA

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nifH Promoter Activity Is Regulated by DNA Supercoiling in Sinorhizobium meliloti

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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 221-226页

作者： Yan-Jie LIU Biao HU Jia-Bi ZHU Shan-Jiong SHEN Guan-Qiao YU National laboratory of plant molecular genetics.institute of plant physiology and ecology,shanghai institutes for biological sciences.chinese academy of sciences,shanghai 200032.china National laboratory of plant molecular genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China

In prokaryotes,DNA supercoiling regulates the expression of many genes;for example,theexpression of Klebsiella pneumoniae nifLA operon depends on DNA negative supercoiling in anaerobicallygrown cells,which indicates that DNA supercoiling might play a role in gene regulation of the *** the expression of the nifH promoter in Sinorhizobium meliloti is not repressed by oxygen,itis proposed that the status of DNA supercoiling may not affect the expression of the nifH *** this hypothesis by analyzing nifH promoter activity in wild-type and gyr Escherichia coli in thepresence and absence of DNA gyrase *** results show that gene expression driven by the *** nifH promoter requires the presence of active DNA *** DNA gyrase increases thenumber of negative superhelical turns in DNA in the presence of ATP,our data indicate that negativesupercoiling is also important for nifH promoter *** study also shows that the DNA supercoiling-dependent *** nifH promoter activity is related to the trans-acting factors NtrC and NifA that *** supercoiling appeared to have a stronger effect on NtrC-activated nifH promoter activity than onNifA-activated promoter ***,these results from the *** nifH promoter model systemseem to indicate that,in addition to regulating gene expression during anaerobic signaling,DNA supercoilingmay also provide a favorable topology for trans-acting factor binding and promoter activation regardless ofoxygen status.

关键词： nifH DNA supercoiling gyrase Sinorhizobium meliloti gene expression

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Substitutions of the Conserved Gly47 Affect the CF1 Inhibitor and Proton Gate Functions of the Chloroplast ATP Synthase ε Subunit

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Acta Biochimica et Biophysica Sinica 2005年第7期37卷 453-462页

作者： Hui DONG Zhang-Lin NI Jia-Mian WEI institute of plant physiology & ecology,shanghai institutes for biological sciences.chinese academy of sciences,shanghai 200032.china institute of plant physiology & ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

The conserved residue Gly47 of the chloroplast ATP synthase ε subunit was substituted withLeu,Arg,Ala and Glu by site-directed *** process generated the mutants εG47L,εG47R,εG47A and εG47E,*** the ε variants showed lower inhibitory effects on the soluble CF1（-ε）Ca2+-ATPase compared with wild-type ε.In reduced conditions,εG47E and εG47R had a lower inhibitoryeffect on the oxidized CF1（-ε）Ca2+-ATPase compared with wild-type ε.In contrast,εG47L and εG47Aincreased the Ca2+-ATPase activity of soluble oxidized CF1（-ε）.The replacement of Gly47 significantlyimpaired the interaction between the subunit ε and γ in an in vitro binding *** study showed thatall ε variants were more effective in blocking proton leakage from the thylakoid *** enhancedATP synthesis of the chloroplast and restored ATP synthesis activity of the reconstituted membranes to alevel that was more efficient than that achieved by wild-type ε.These results indicate that the conservedGly47 residue of the ε subunit is very important for maintaining the structure and function of the ε subunitand may affect the interaction between the ε subunit,β subunit of CF1and subunit Ⅲ of CF_o,therebyregulating the ATP hydrolysis and synthesis,as well as the proton translocation role of the subunit Ⅲ of CF_o.

关键词： chloroplast ATP synthase ε subunit GST pull-down assay site-directed mutagenesis

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Are small RNAs a big help to plants?

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Science china(Life sciences) 2009年第3期52卷 212-223页

作者： MAO YingBo,XUE XueYi & CHEN XiaoYa National key laboratory of plant molecular genetics.institute of plant physiology and ecology,shanghai institutes for biological sciences.chinese academy of sciences,shanghai 200032.china National key laboratory of plant molecular genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

The discovery of RNA interference(RNAi) has augmented our knowledge of gene regulation and presents a fascinating technology that has a great potential for application in genetic analysis,disease therapy,plant protection,and many other *** this review,we will focus on the biological functions of RNAi and its application in agriculture with a brief introduction to the history of its discovery and molecular mechanism.

关键词： RNA interference gene regulation plant protection

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Agrobacterium tumefaciens-mediated transformation of rice with the spider insecticidal gene conferring resistance to leaffolder and striped stem borer

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Cell Research 2001年第2期11卷 149-155页

作者： HUANG JIAN QIU, ZHI MING WEI, HAI LONG AN, YU XIAN ZHU National laboratory of plant molecular genetics, institute of plant physiology and ecology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200032, china National La National laboratory of plant molecular genetics Institute of Plant Physiology and Ecology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Shanghai China National laboratory of plant Genetic Engineering and Protein Engineering College of Life Science Peking University Beijing China

Immature embryos of rice varieties 'Xiushui11' and 'Chunjiang 11' precultured for 4d were infected and transformed by Agrobacterium tumefaciens strain EHA101/pExT7 (containing the spider insecticidal gene). The resistant cant were transferred onto the differentiation medium and plants were regenerated. The transformation frequency reached 56%-72% measured as numbers of Geneticin (G418)-resistant calli produced and 36%-60% measured as numbers of transgenic plants regenerated, respectively. PCR and Southern blot analysis of transgenic plants confirmed that the T-DNA had been integrated into the rice genome. Insect bioassays using T1 transgenic plants indicated that the mortality of the leaffolder (Cnaphalocrasis medinalis) after 7d of leaf feeding reached 38%-61% and the corrected mortality of the striped stem borer (Chilo suppressalis) after 7d of leaf feeding reached 16%-75%. The insect bioassay results demonstrated that the transgenic plants expressing the spider insecticidal protein conferred enhanced resistance to these pests.

关键词： Rice Agrobacterium tumefaciens spider insecticidal gene transgenic plant Leaffolder striped stem borer insect bioassay.

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Regeneration of plants from callus tissues of hairy roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi

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Cell Research 2001年第4期11卷 279-284页

作者： WANG YU MEI, JIANG BO WANG, DA LUO , JING FEN JIA National laboratory of plant molecular genetics, institute of plant physiology and ecology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200032, china Department of Bi 中科院上海植物生理生态所上海 200032 西北大学生物系陕西西安 710069

The legume forage Alhagi pseudoalhagi was transformed by the Agrobacterium rhizogenes strain A4 using cotyledon and hypocotyl segments as infection materials. Regenerated plants were achieved from sterile calli derived from hairy roots, which occurred at or near the infection sites. The regenerated plants from hairy root were characterized by normal leaf morphology and stem growth but a shallow and more extensive root system than normal plants. Opine synthesis, PCR and Southern blot confirmed that T- DNA had been integrated into the A. pseudoalhagi genome. Acetosyringone (AS) was found to be vital for successful transformation of A. pseudoalhagi.

关键词： Alhagi pseudoalhagi Agrobacterium rhizogenes hairy root regeneration.

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Sequencing and Analysis of a Genomic Fragment Provide an Insight into the Dunaliella viridis Genomic Sequence

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 812-820页

作者： Xiao-Ming SUN~(1#) Yuan-Ping TANG~(1#) Xiang-Zong MENG~1 Wen-Wen ZHANG~1 Shan LI~1 Zhi-Rui DENG~1 Zheng-Kai XU~(1,2) Ren-Tao SONG~1* 1 shanghai key laboratory of Bio-energy Crops,School of Life sciences,shanghai University,shanghai 200444,china 2 institute of plant physiology & ecology,shanghai institutes for biological sciences.chinese academy of sciences,shanghai 200032.china shanghai key laboratory of Bio-energy Crops School of Life Sciences Shanghai UniversityShanghai 200444 China institute of plant physiology & ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

Dunaliella is a genus of wall-less unicellular eukaryotic green *** exceptional resistancesto salt and various other stresses have made it an ideal model for stress tolerance ***,very littleis known about its genome and genomic *** this study,we sequenced and analyzed a 29,268 bpgenomic fragment from DunalieIla *** fragment showed low sequence homology to the *** the nucleotide level,only a segment with significant sequence homology to 18S rRNA *** fragment contained six putative genes,but only one gene showed significant homology at theprotein level to GenBank *** average GC content of this sequence was 51.1%,which was muchlower than that of close related green algae Chlamydomonas （65.7%）.Significant segmental duplicationswere found within this *** duplicated sequences accounted for about 35.7% of the *** amounts of simple sequence repeats （microsatellites） were found,with strong bias towards（AC）_n type （76%）.Analysis of other Dunaliella genomic sequences in the GenBank database （total 25,749bp） was in agreement with these *** sequence features made it difficult to sequence Dunaliellagenomic *** investigation should be made to reveal the biological significance of these uniquesequence features.

关键词： Dunaliella viridis genomic sequence sequence feature sequence duplication simple sequence repeat

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