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Cell Research 2005年第8期15卷 585-592页

作者： Jing JIANG, Ling Wen FAN, Wei Hua WU* The state key laboratory of plant physiology & biochemistry, college of biological sciences, china agricultural university, beijing 100094, china The state key laboratory of plant physiology & biochemistry College of Biological Sciences China Agricultural University Beijing China

Although there were reports suggesting the involvement of endogenous cAMP in plant defense signaling cascades, there is no direct evidence supporting this notion yet and the detailed mechanism is unclear. In the present study, we have used pathogenic fungi Verticillium dahliae and Arabidopsis plants as a model system of plant-microb interaction to demonstrate the function of endogenous cAMP in Arabidopsis defense responses. Both V. dahliae inoculation and Verticillium toxins injection induced typical “wilt” symptoms in Arabidopsis seedlings. When either 8-Br-AMP (a membrane permeable cAMP analogue) or salicylic acid (SA) was applied to Arabidopsis, the plants became resistant to V. dahliae toxins. However, addition of 8-Br-AMP did not increase the resistance of Arabidopsis transgenic plants deficient in SA to the toxins, suggesting that cAMP might act upstream of SA in plant defense signaling pathway. Indeed, 8-Br-cAMP and forskolin, an activator of adenylyl cyclase, significantly stimulated the endogenous SA level in plants, whereas DDA, an inhibitor of adenylyl cyclase dramatically reduced toxin-induced SA increase. Both the endog- enous cAMP and SA increased significantly in Arabidopsis seedlings treated with toxins. Furthermore, transcription level of pathogenesis-related protein 1 gene (PR1) was strongly induced by both 8-Br-cAMP and the toxin treatment. Taken together, our data demonstrate that endogenous cAMP is involved in plant defense responses against Verticillium- secreted toxins by regulating the production of the known signal SA in plant defense pathway.

关键词： Arabidopsis thaliana Verticillium dahliae cyclic AMP alicylic acid signal transduction defense response.

Ca^(2+)-independent ZmCPK2 is inhibited by Ca^(2+)-dependent ZmCPK17 during drought response in maize

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Journal of Integrative plant Biology 2024年第7期66卷 1313-1333页

作者： Xiaoying Hu Jinkui Cheng Minmin Lu Tingting Fang Yujuan Zhu zhen Li Xiqing Wang Yu Wang yan guo Shuhua yang Zhizhong Gong state key laboratory of plant Environmental Resilience Frontiers Science Center for Molecular Design BreedingCenter for Crop Functional Genomics and Molecular BreedingCollege of Biological SciencesChina Agricultural UniversityBeijing 100193China college of Life sciences Institute of Life Science and Green DevelopmentHebei UniversityBaoding 071002China

Calcium oscillations are induced by different ***-dependent protein kinases(CDPKs/CPKs)are one major group of the plant calcium decoders that are involved in various processes including drought *** CPKs are ***,we identified ZmCPK2 as a negative regulator of drought resistance by screening an overexpression transgenic maize *** found that ZmCPK2 does not bind calcium,and its activity is mainly inhibited during short term abscisic acid(ABA)treatment,and dynamically changed in prolonged ***,ZmCPK2 interacts with and is inhibited by calcium-dependent ZmCPK17,a positive regulator of drought resistance,which is activated by ***17 could prevent the nuclear localization of ZmCPK2 through phosphorylation of ***2 interacts with and phosphorylates and activates ZmYAB15,a negative transcriptional factor for drought *** results suggest that drought stress-induced Ca^(2+)can be decoded directly by ZmCPK17 that inhibits ZmCPK2,thereby promoting plant adaptation to water deficit.

关键词： CDPK drought response phosphorylation stomatal movement transcriptional regulation

The light and hypoxia induced gene ZmPORB1 determines tocopherol content in the maize kernel

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Science china(Life sciences) 2024年第3期67卷 435-448页

作者： Nannan Liu Yuanhao Du Shijuan yan Wei Chen Min Deng Shutu Xu Hong Wang Wei Zhan Wenjie Huang yan Yin Xiaohong yang Qiao zhao Alisdair R.Fernie Jianbing yan National key laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070China Hubei Hongshan laboratory Wuhan 430070China Agro-biological Gene Research Center Guangdong Academy of Agricultural SciencesGuangzhou 510640China college of Agronomy Hunan Agricultural UniversityChangsha 410128China college of Agronomy Northwest A&F UniversityXi'an 710000China state key laboratory of North china Crop Improvement and Regulation Hebei Sub-center of National Maize Improvement Center of ChinaCollege of AgronomyHebei Agricultural UniversityBaoding 071001China college of Life sciences South-Central Minzu UniversityWuhan 430070China plant Science Facility of the Institute of Botany Chinese Academy of SciencesBeijing 100093China National Maize Improvement Center of china Beijing Key Laboratory of Crop Genetic ImprovementChina Agricultural UniversityBeijing 100093China Shenzhen key laboratory of Synthetic Genomics Guangdong Provincial Key Laboratory of Synthetic GenomicsShenzhen Institute of Synthetic BiologyShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen 518055China Max Planck Institute of Molecular plant physiology Potsdam-Golm 14476Germany

Tocopherol is an important lipid-soluble antioxidant beneficial for both human health and plant growth. Here, we fine mapped a major QTLqVE1 affecting γ-tocopherol content in maize kernel, positionally cloned and confirmed the underlying gene ZmPORB1(por1), as a protochlorophyllide oxidoreductase. A 13.7 kb insertion reduced the tocopherol and chlorophyll content, and the photosynthetic activity by repressing ZmPORB1 expression in embryos of NIL-K22, but did not affect the levels of the tocopherol precursors HGA(homogentisic acid)and PMP(phytyl monophosphate). Furthermore, ZmPORB1 is inducible by low oxygen and light, thereby involved in the hypoxia response in developing embryos. Concurrent with natural hypoxia in embryos, the redox state has been changed with NO increasing and H_(2)O_(2) decreasing, which lowered γ-tocopherol content via scavenging reactive nitrogen species. In conclusion, we proposed that the lower lightharvesting chlorophyll content weakened embryo photosynthesis, leading to fewer oxygen supplies and consequently diverse hypoxic responses including an elevated γ-tocopherol consumption. Our findings shed light on the mechanism for fine-tuning endogenous oxygen concentration in the maize embryo through a novel feedback pathway involving the light and low oxygen regulation of ZmPORB1 expression and chlorophyll content.

关键词： maize tocopherol hypoxia photosynthesis ZmPORB1

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Overexpression of the tonoplast aquaporin AtTIP5;1 conferred tolerance to boron toxicity in Arabidopsis

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Journal of Genetics and Genomics 2010年第6期37卷 389-397页

作者： Yongqi pang, Lijuan Li, Fei Ren, Pingli Lu, Pengcheng Wei, Jinghui Cai, Lingguo Xin, Juan Zhang, Jia Chen, Xuechen Wang state key laboratory of plant physiology and biochemistry, college of biological sciences, china agricultural university, beijing 100193, china state key laboratory of plant physiology and biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China

Boron （B） toxicity to plants is responsible for low crop productivity in many regions of the world. Here we report a novel and effective means to alleviate the B toxicity to plants under high B circumstance. Functional characterization of AtTIP5;1, an aquaporin gene, re- vealed that overexpression ofAtTIP5;1 （OxAtT1P5;1） in Arabidopsis significantly increased its tolerance to high B toxicity. Compared to wild-type plants, OxAtTIP5;1 plants exhibited longer hypocotyls, accelerated development, increased silique production under high B treatments. GUS staining and quantitative RT-PCR （qRT-PCR） results demonstrated that the expression of AtTIP5;1 was induced by high B concentration treatment. Subcellular localization analysis revealed that the AtTIP5;1-GFP fusion protein was localized on the tonoplast membrane, which was consistent with the prediction based on bioinformatics. Taken together, our results suggest that AtTIP5; 1 is in- volved in B transport pathway possibly via vacuolar compartmentation for B, and that overexpression of AtTIP5;1 in plants may provide an effective way to overcome the problem resulting from high B concentration toxicity.

关键词： aquaporin boron toxicity tolerance

A Genome-wide Functional Characterization of Arabidopsis Regulatory Calcium Sensors in Pollen Tubes

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Journal of Integrative plant Biology 2009年第8期51卷 751-761页

作者： Liming Zhou1,2, Ying Fu1,2 and zhenbiao yang2,3 (1state key laboratory of plant physiology and biochemistry, Department of plant sciences, college of biological sciences, china agricultural university, beijing 100193, china 2china agricultural university (CAU)-university of California, Riverside (UCR) Joint Center for biological sciences and Biotechnology, college of biological sciences, china agricultural university, beijing 100193, china 3Center for plant Cell Biology and Department of Botany and plant sciences, university of California, Riverside, CA 92521, USA) state key laboratory of plant physiology and biochemistry Department of Plant Sciences College of Biological Sciences China Agricultural University Beijing 100193 China china agricultural university (CAU)-university of California Riverside (UCR) Joint Center for Biological Sciences and Biotechnology College of Biological Sciences China Agricultural University Beijing 100193 China Center for plant Cell Biology and Department of Botany and plant sciences University of California Riverside CA 92521 USA)

Calcium, an ubiquitous second messenger, plays an essential and versatile role in cellular signaling. The diverse function of calcium signals is achieved by an excess of calcium sensors. plants possess large numbers of calcium sensors, most of which have not been functionally characterized. To identify physiologically relevant calcium sensors in a specific cell type, we conducted a genome-wide functional survey in pollen tubes, for which spatiotemporal calcium signals are well-characterized and required for polarized tip growth. *** members of calrnodulin （CAM）, CaM-like （CML）, calcium-dependent protein kinase （CDPK） and calcineurin B-like protein （CBL） families were tagged with green fluorescence protein （GFP） and their localization patterns and overexpression phenotypes were characterized in tobacco pollen tubes. We found that several fusion proteins showed distinct overexpression phenotypes and subcellular localization patterns. CDPK24-GFP was localized to the vegetative nucleus and the generative cell/sperms. CDPK32-GFP caused severe growth depolarization. CBL2-GFP and CBL3-GFP exhibited dynamic patterns of subcellular localization, including several endomembrane compartments, the apical plasma membrane （PM）, and cytoskeleton-like structures in pollen tubes. Their overexpression also inhibited pollen tube elongation and induced growth depolarization. These putative calcium sensors are excellent candidates for the calcium sensors responsible for the regulation of calcium homeostasis and calciumdependent tip growth and growth oscillation in pollen tubes.

关键词： Arabidopsis calcium sensor calcium signaling Rho-related GTPase from plants subcellular localization tip growth

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.

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.

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

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 state key laboratory of plant physiology and biochemistry College of Biological Sciences China Agricultural University Beijing China college of Life Science and Technology Guangxi University Nanning 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