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Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants: silencing of the tomato ferric chelate reductase gene, FRO1, as an example

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science china(life sciences) 2008年第5期51卷 402-409页

作者： HE XiuXia1,2,JIN ChongWei1, LI GuiXin3, YOU GuangYi4, ZHOU XuePing3 & ZHENG ShaoJian4 1 key laboratory for Environmental and Ecosystem Health of the Ministry of Education, college of Environmental and Resource science, zhejiang university, hangzhou 310029, china 2 college of life science, Changchun university of science and Technology, Changchun 130022, china 3 Institute of Biotechnology, zhejiang university, hangzhou 310029, china 4 state key laboratory of plant physiology and biochemistry, college of life science, zhejiang university, hangzhou 310058, china 1. key laboratory for Environmental and Ecosystem Health of the Ministry of Education College of Environmental and Resource Science Zhejiang University Hangzhou 310029 China 2. college of life science Changchun University of Science and Technology Changchun 130022 China 3. Institute of Biotechnology Zhejiang University Hangzhou 310029 China 4. state key laboratory of plant physiology and biochemistry College of Life Science Zhejiang University Hangzhou 310058 China

Virus-induced gene silencing (VIGS) is potentially an attractive reverse-genetics tool for studies of plant gene function, but whether it is effective in silencing mineral nutritional-related genes in roots has not been demonstrated. Here we report on an efficient VIGS system that functions in tomato roots using a modified viral satellite DNA (DNAmβ) associated with Tomato yellow leaf curl china virus (TYLCCNV). A cDNA fragment of the ferric chelate reductase gene (FRO1) from tomato was inserted into the DNAmβ vector. Tomato roots agro-inoculated with DNAmβ carrying both a fragment of FRO1 and TYLCCNV used as a helper virus exhibited a significant reduction at the FRO1 mRNA level. As a consequence, ferric chelate reductase activity, as determined by visualization of the pink FeBPDS3 complex was significantly decreased. Our results clearly demonstrated that VIGS system can be employed to investigate gene function associated with plant nutrient uptake in roots.

关键词： virus-induced gene silencing roots ferric chelate reductase

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Comparing Genetic Characteristics of Retrotransposon TOS17 During Different Tissue Culture Processes in the Rice Cultivars Nipponbare and Shishoubaimao

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Agricultural sciences in china 2010年第2期9卷 157-162页

作者： LI Kuan, WANG Chuang, YING Shan and SHOU Hui-xia state key laboratory of plant physiology and biochemistry, college of life sciences, zhejiang university, hangzhou 310058, P.R.china [a] state key laboratory of plant physiology and biochemistry College of Life Sciences Zhejiang University Hangzhou 310058 P.R. China

As a retrotransposon, TOS17 was a useful tool for rice genetic and functional genomic research. To ascertain the feasibility of constructing a TOS17 insertion mutation library in the rice cultivar Shishoubaimao, the genetic and expression characteristics of TOS17 were analyzed. We made solid and suspension tissue cultures and confirmed the copy numbers of TOS17 at different time points in both tissue culture processes by real-time quantitative PCR （RT-qPCR）. Three primary copies of TOS17 were detected in naturally grown Shishoubaimao. TOS17 was activated by tissue culture, and the copy numbers of TOSI7 increased along with a prolonged tissue culture time in both the Nipponbare and the Shishoubaimao cultivars. Therefore, Shishoubaimao is a potential candidate for constructing a TOS17 insertion mutant library. Compared with Nipponbare, TOS17 was more active in Shishoubaimao during tissue culture. Higher copy numbers of TOS17 were obtained with the suspension tissue culture process than with the solid tissue culture process over the same time courses. We concluded that 3-4 months of suspension tissue culture time is suitable for constructing a TOS17 insertion mutant library in Shishoubaimao.

关键词： Shishoubaimao tissue culture TOS17 copy number insertion mutant library

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Involvement of NADPH oxidase NtrbohD in the rapid production of H2O2 induced by ABA in cultured tobacco cell line BY-2

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Progress in Natural science:Materials International 2008年第3期18卷 267-271页

作者： Fushun Hao a,b, Jinguang Zhang a, Zhonglian Yu a, Jia Chen a, a state key laboratory of plant physiology and biochemistry, college of Biological science, china Agricultural university, Beijing 100094, china b Henan key laboratory of plant Stress Biology, college of life science, Henan university, Kaifeng 475004, china state key laboratory of plant physiology and biochemistry College of Biological Science China Agricultural University Beijing 100094 China Henan key laboratory of plant Stress Biology College of Life Science Henan University Kaifeng 475004 China

The mechanisms for the production of hydrogen peroxide (H2O2) induced by abscisic acid (ABA) were investigated in suspension culture cells of tobacco BY-2 cells. The results showed that the immediate generation of H2O2, which was mainly derived from super- oxide dismutase-catalyzed dismutation of superoxide radical, was significantly induced by ABA. Furthermore, treatment of the cultured tobacco cells with ABA resulted in a time-dependent quick increase in plasma membrane (PM) NADPH oxidase activity, which coin- cided on time and magnitude with the elevation in ABA-induced accumulation of H2O2. Moreover, these enhanced effects were pro- nouncedly inhibited by two NADPH oxidase inhibitors, diphenylene iodonium and imidazole, suggesting that PM NADPH oxidase is involved in the rapid accumulation of H2O2 in cultured tobacco cells. In addition, analysis of the expression level of NtrbohD, a PM NADPH oxidase gene in tobacco, by RT-PCR and protein gel blot revealed that the gene at both mRNA and protein levels was upregulated by ABA, indicating that NtrbohD participates in the ABA-stimulated rapid production of H2O2 in tobacco culture cells. Taken together, these findings suggest that ABA induces the rapid accumulation of reactive oxygen species via NADPH oxidase in sus-pension culture cells of tobacco, and that NADPH oxidase and H2O2 appear to be important components in ABA signal transduction pathway in plants.

关键词： ABA Cultured tobacco cells H2O2 NADPH oxidase NtrbohD

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Isolation and expression of an aquaporin-like gene VfPIP1 in Vicia faba

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Progress in Natural science:Materials International 2005年第6期15卷 496-501页

作者： CUI Xianghuan, HAO Fushun, CHEN Hui, CAI Jinghui, CHEN Jia and WANG Xuechen*(state key laboratory for plant physiology and biochemistry, college of Biological science, china Agricultural university, Beijing 100094, china) state key laboratory for plant physiology and biochemistry College of Biological Science China Agricultural University Beijing 100094 China

To explore the effects of aquaporins on stomatal movement, we isolated a full length cDNA of aquaporin-like gene VfPIP1 (Vicia faba plasma membrane intrinsic protein gene, GenBank accession number: AY667436), which encodes for a 290-amino-acid polypeptide, from Vicia faba leaf epidermis by 5′/3′ RACE (Rapid Amplification of cDNA Ends). The analyses of VfPIP1 transmembrane regions and amino acid sequence show that VfPIP1 owns six membrane-spanning domains and the special plasma membrane signature sequences GGGANXXXXGY and TGI/TNPARSL/FGAAI/VI/VF/YN, and it should be a member of PIP1 subfamily. The results of in situ hybridization and Northern blot indicate that VfPIP1 is strongly expressed in guard cells and induced by ABA. Hereby, VfPIP1 may be involved in the water-transmembrane movement of guard cells.

关键词： ABA aquaporin gene cloning Vicia faba guard cell.

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Soluble expression and characterization of a GFP-fused pea actin isoform (PEAc1)

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Cell Research 2004年第5期14卷 407-414页

作者： Ai Xiao LIU*, Shao Bin ZHANG*, Xiao Jing XU, Dong Tao REN, Guo Qin LIU** state key laboratory of plant physiology and biochemistry, college of Biological science, china Agricultural university, Beijing 100094, china. state key laboratory of plant physiology and biochemistry College of Biological Science China Agricultural University Beijing 100094 China.

A pea actin isoform PEAc1 with green fluorescent protein (GFP) fusion to its C-terminus and His-tag to its N- terminus, was expressed in prokaryotic cells in soluble form, and highly purified with Ni-Chelating SepharoseTM Fast Flow column. The purified fusion protein (PEAc1-GFP) efficiently inhibited DNase I activities before polymerization, and activated the myosin Mg-ATPase activities after polymerization. The PEAc1-GFP also polymerized into green fluorescent filamentous structures with a critical concentration of 0.75 μM. These filamentous structures were labeled by TRITC-phalloidin, a specific agent for staining actin microfilaments, and identified as having 9 nm diameters by negative staining. These results indicated that PEAc1 preserved the essential characteristics of actin even with His-tag and GFP fusion, suggesting a promising potential to use GFP fusion protein in obtainning soluble plant actin isoform to analyze its physical and biochemical properties in vitro. The PEAc1-GFP was also expressed in tobacco BY2 cells, which offers a new pathway for further studying its distribution and function in vivo.

关键词： actin isoform polymerization DNase I inhibition myosin Mg-ATPase activation expression.

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A membrane-tethered transcription factor ANAC089 negatively regulates floral initiation in Arabidopsis thaliana

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science china(life sciences) 2010年第11期53卷 1299-1306页

作者： LI JianQin1,2, ZHANG Juan1, WANG XueChen1 & CHEN Jia1 1 state key laboratory of plant physiology and biochemistry,college of Biological sciences, china Agricultural university, Beijing 100193, china 2 college of life science and Technology, Guangxi university, Nanning 530005, china 1. state key laboratory of plant physiology and biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China2. College of Life Science and Technology Guangxi University Nanning 530005 China

The plant-specific NAC (NAM, ATAF1/2,and CUC2) transcription factors have a regulatory function in developmental processes and stress responses. Notably a group of NAC members named NTLs (NTM1-Like) are membrane-tethered, ensuring plants rapidly respond to developmental changes and environmental stimuli. Our results indicated that ANAC089 was a membrane-tethered transcription factor and its truncated form was responsible for the physiological function in flowering time control.

关键词： Arabidopsis thaliana ANAC089 floral initiation membrane-tethered transcription factor

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Computational Identification of Protein-Protein Interactions in Rice Based on the Predicted Rice Interactome Network

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Genomics,Proteomics & Bioinformatics 2011年第Z2期 128-137页

作者： Pengcheng Zhu1, Haibin Gu1, Yinming Jiao1, Donglin Huang1,2, and Ming Chen1,2,3 1Department of Bioinformatics, college of life sciences, zhejiang university, hangzhou 310058, china 2James D. Watson Institute of Genome sciences, zhejiang university, hangzhou 310058, china 3state key laboratory of plant physiology and biochemistry, college of life sciences, zhejiang university, hangzhou 310058, china

plant protein-protein interaction networks have not been identified by large-scale experiments. In order to better understand the protein interactions in rice, the Predicted Rice Interactome Network (PRIN; http://***/ prin/) presented 76,585 predicted interactions involving 5,049 rice proteins. After mapping genomic features of rice (GO annotation, subcellular localization prediction, and gene expression), we found that a well-annotated and biologically significant network is rich enough to capture many significant functional linkages within higher-order biological systems, such as pathways and biological processes. Furthermore, we took MADS-box do- main-containing proteins and circadian rhythm signaling pathways as examples to demonstrate that functional protein complexes and biological pathways could be effectively expanded in our predicted network. The expanded molecular network in PRIN has considerably improved the capability of these analyses to integrate existing knowledge and provide novel insights into the function and coordination of genes and gene networks.

关键词： protein-protein interactions rice interactome interolog sub-network expansion pathway clustering

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OslAA23-mediated auxin signaling defines postembryonic maintenance of QC in primary roots and lateral and adventitious root development in rice

OslAA23-mediated auxin signaling defines postembryonic maint...

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第十一届全国植物基因组学大会

作者： Jun Ni,Gaohang Wang,Ping Wu~* state key laboratory of plant physiology and biochemistry,college of life science,zhejiang university, hangzhou 310058,P.R.china

Here we report on a member of Aux/IAA family in rice,OsIAA23,for postembryonic maintenance of quiescent center(QC) and stem cells,and the lateral root and adventitious root *** stabilizing of OsIAA23 results in defect...

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Effects of the disaggregation of high-polymerized particles in guard cell vacuoles on osmoregulation of stomatal aperture (stomata opening)

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Chinese science Bulletin 2003年第5期48卷 453-456页

作者： YE Qing, ZHU Guoli & LOU Chenghou Biology college, china Agricultural university, state key laboratory of plant physiology and biochemistry, Beijing 100094, china Correspondence should be addressed to Zhu Guoli (e-mail: glzhu@mail. ***) Biology college China Agricultural University State Key Laboratory of Plant Physiology and Biochemistry

Observation under an electron microscope reveals that in closed and open stomata of V. faba, the aver-age volume of particles in guard cell vacuoles (GCV) reduces about 3 orders in magnitude, while the distribution density of the particles increases about 2 orders of magnitude. By using the method of the ratio of fluorescent emissions with laser scanning confocal microscopy, the monitoring to sto-mata opening shows that during 10 to 30 s before the first distinguishable aperture of stomata, there is a change of pH in GCV about -0.5 units. A quick stomatal opening immedi-ately follows the changes of pH in GCV to reach a steady aperture about 12 mm in 100200 s. This work proposes a model for the osmoregulation in GCV for stomatal opening. The proposed osmoregulation is related to the disaggregation of some polymerized particles inside GCV, which is probably induced by a -DpH in the vacuole. This model describes a process of osmoregulation that avoids the massive energy consuming transportation across cell membranes, which is a foundation of the current chemiosmotic hypothesis. This model is a supplement to the multiple controlling hypothesis for the stomatal movement, which widens research principle ideas for other quick movements in plants.

关键词：气孔运动气孔开放保卫细胞液泡渗透调节高聚合微粒解体

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Two Rubisco Activase Isoforms May Play Different Role in Photosynthetic Heat Acclimation in Rice plant

Two Rubisco Activase Isoforms May Play Different Role in Pho...

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中国植物生理学会第十次会员代表大会暨全国学术年会

作者： Dun Wang, Xiaofei Li, Wanjun Yang, Xuping Feng, Dean Jiang* state key laboratory of plant physiology and biochemistry, college of life sciences, zhejiang university, hangzhou 310058, china

＜正＞Studies in many species have shown that increasing growth temperature gradually or pre-treatment can lead

关键词： Rubisco activase isoforms heat acclimation photosynthetic rate

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