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Analysis of the essential DNA region for OsEBP-89 promoter in response to methyl jasmonic acid

Science china(life sciences) 2008年第3期51卷 280-285页

作者： LI Ang1,2, CHEN LiangLiang1,3, REN HaiYun3, WANG XueChen2, ZHANG HaiWen1,4 & HUANG Rong- Feng1,4 1 Biotechnology Research Institute, Chinese Academy of agricultural sciences, beijing 100081, china 2 national laboratory of plant physiology and biochemistry, college of biological sciences, china agricultural university, beijing 100094, china 3 college of life sciences, beijing normal university, beijing 100875, china 4 national Key Facility of Crop Gene Resources and Genetic Improvement, beijing 100081, china Biotechnology Research Institute Chinese Academy of Agricultural Sciences Beijing 100081 China national laboratory of plant physiology biochemistry College of Biological Sciences China Agricultural University Beijing 100094 China college of life sciences Beijing Normal University Beijing 100875 China national Key Facility of Crop Gene Resources Genetic Improvement Beijing 100081 China

In rice, the characterization of OsEBP-89 is inducible by various stress- or hormone-stimuli, including ethylene, abscisic acid (ABA), jasmonate acid (JA), drought and cold. Here, we report the investigation of essential DNA region within OsEBP-89 promoter for methyl jasmonic acid (MeJA) induction. PLACE analysis indicates that this promoter sequence contains multiple potential elements in response to various stimuli. First, we fused this promoter with GUS gene and analyzed its expression under MeJA treatment through Agrobacterium infiltration mediating transient expression in tobacco leaves. Our results revealed that this chimeric gene could be inducible by MeJA in tobacco leaves. To further de- termine the crucial sequences responsible for MeJA induction, we generated a series of deletion pro- moters which were fused with GUS reporter gene respectively. The results of transient expression of GUS gene driven by these mutant promoters show that the essential region for MeJA induction is po- sitioned in the region between -1200 and -800 in OsEBP-89 promoter containing a G-box (?1127), which is distinct from the essential region containing ERE (?562) for ACC induction. In all, our finding is helpful in understanding the molecular mechanism of OsEBP-89 expression under different stimuli.

关键词： OsEBP-89 essential DNA region methyl jasmonic acid transient assay promoter tobacco leaves

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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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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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Actomyosin is Involved in the Organization of the Microtubule Preprophase Band in Arabidopsis Suspension Cultured Cells

Actomyosin is Involved in the Organization of the Microtubul...

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第四届拟南芥学术研讨会

作者： Chun-Li Li,Zhi-Ling Chen and Ming Yuan State Key laboratory of plant physiology and biochemistry,Department of plant sciences,college of biological sciences,china agricultural university,beijing 100094,china college of life sciences,Capital normal university,beijing 100037,china

The microtubule preprophase bands (PPBs) participate in the sequence of events to position cell plates in most plants. However, the mechanism of PPB formation remains to be clarified. In the present study, the organization of PPBs in Arabidopsis suspension cultured cells was investigated by confocal laser scanning microscopy combined with pharmacological treatments of reagents specific for the cytoskeleton elements. Double staining of F-actin and microtubules (MTs) showed that actin filaments were arranged randomly and no coiocaiization with cortical MTs was observed in the interphase cells. However, cortical actin filaments showed coiocaiization with MTs during the formation of PPBs. A broad actin band formed with the broad MT band in the initiation of PPB and narrowed down together with the MT band to form the PPB. Nevertheless, broad MT bands were formed but failed to narrow down in cells treated with the F-actin disruptor latrunculin A. In contrast, in the presence of the F-actin stabilizer phalloidin, PPB formation did not exhibit any abnormality. Therefore, the integrity, but not the dynamics, of the actin cytoskeleton is necessary for the formation of normal PPBs. Treatment with 2,3-butanedine monoxime, a myosin inhibitor, also resulted in the formation of broad MT bands, indicating that actomyosin may be involved in the rearrangement of MTs to form the PPBs. Double staining of MTs and myosin revealed that myosin concentrated on the PPB region during PPB formation. It is suggested that the actin cytoskeleton at the PPB site may serve as a rack to transport cortical MTs by using myosin when the broad MT band narrows down to form the PPB.

关键词： Arabidopsis suspension cultured cell 2,3-butanedine monoxime microfilament microtubule preprophase band myosin

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Cloning and functional analysis of chloroplast division gene NtFtsZ2-l in Nicotiana tabacum

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Progress in Natural Science:Materials International 2003年第5期 39-43页

作者： KONG Dongdong , JU Chuanli, WANG Dong, HU Yong, ZHU Zhiqing, HE Yikun and SUN Jingsan (1. Key laboratory of Photosynthesis and Enviromental Molecular physiology, Institute of Botany, Chinese Academy of sciences, beijing 100093, china 2. Department of Biology, Capital normal university, beijing 100037, china 3. laboratory of Agronomy and Biotechnology, china agricultural university, beijing 100037, china 4. college of life sciences, beijing normal university, beijing 100875, china)

FtsZ protein plays an important role in the division of chloroplasts. With the finding and functional analysis of higher plant FtsZ proteins, people have deepened the understanding in the molecular mechanism of chloroplast division. Multiple ftsZ genes are diversified into two families in higher plants, ftsZ1 and ftsZ2 . On the basis of the research on ftsZl family, we analyzed the function of NtFtsZ2-l gene in Nicotiana tabacum . Microscopic analysis of the sense and antisense NtFtsZ2-l transgenic tobacco plants revealed that the chloroplasts were abnormal in size and also in number when compared with wild-type tobacco chloroplasts. Our investigations confirmed that the NtFtsZ2-l gene is involved in plant chloroplast division.

关键词： Nicotiana tabacum NtFtsZ2-l gene chloroplast division.

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Depolymerization of actin cytoskeleton is involved in stomatal closure-induced by extracellular calmodulin in Arabidopsis

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Science china(life sciences) 2004年第5期47卷 454-460页

作者： XIAO Yumei1, CHEN Yuling1,2, HUANG Rongfeng3, CHEN Jia1 & WANG Xue-chen1 1. The national Key laboratory of plant physiology and biochemistry, college of biological sciences, china agricultural university, Bei-jing 100094, china 2. college of life sciences, Hebei normal university, Shijiazhuang 050016, china 3. Biotechnology Research Institute, Chinese Academy of agricultural sciences, beijing 100081, china 1. The national Key laboratory of plant physiology and biochemistry College of Biological Sciences China Agricultural University 100094 Beijing China 2. college of life sciences Hebei Normal University 050016 Shijiazhuang China 3. Biotechnology Research Institute Chinese Academy of Agricultural Sciences 100081 Beijing China

Extracellular calmodulin (CaM) plays significant roles in many physiological proc-esses,but little is known about its mechanism of regulating stomatal movements. In this paper, whether CaM exists in the guard cell walls of Arabidopsis and whether depolymerization of actin cytoskeleton is involved in extracellular CaM-induced stomatal closing are investigated. It is found that CaM exists in guard cell walls of Arabidopsis, and its molecular weight is about 17 kD. Bioassay using CaM antagonists W7-agarose and anti-CaM serum shows that the endogenous extracellular CaM promotes stomatal closure and delays stomatal opening. The long radial actin filaments in guard cells undergo disruption in a time-dependent manner during exogenous CaM-induced stomatal closing. Pharmacological experiments show that depolymerization of ac-tin cytoskeleton enhances the effect of exogenous CaMinduced stomatal closing and polym-erization reduces the effect. We also find that exogenous CaM triggers an increase in [Ca2+]cyt of guard cells. If [Ca2+]cyt increase is blocked with EGTA, exogenous CaM-induced stomatal closure is inhibited. These results indicate that extracellular CaM causes elevation of [Ca2+]cyt in guard cells, subsequently resulting in disruption of actin filaments and finally leading to guard cells closure.

关键词： Arabidopsis,stomatal movements,extracellular calmodulin,actin,Ca2+.

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Heterotrimeric G protein participated in modulation of cytoplasmic calcium concentration in pollen cells

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Progress in Natural Science:Materials International 2003年第9期 40-44+84页

作者： SHANG Zhonglin, MA Ligeng, WANG Xuechen and SUN Daye (1. The college of biological sciences, Hebei normal university, Shijiazhuang 050016, china 2. State Key laboratory of plant physiology & biochemistry, beijing 100094, china)

Cytoplasmic free calcium concentration([Ca2+]c) in pollen cells of Lilium daviddi is measured with confocal laserscanning microscopy to investigate the effect of heterotrimeric G protein （G protein） on [Ca2+]cand the possible signal transduction path-way of G protein triggering cellular calcium signal. After application, cholera toxin （CTX）, an agonist of G protein, triggers a transientincrease of [Ca2+]cin pollen cells, and evokes a spatial-temporal characteristic calcium dynamics; while pertussis toxin （PTX）, a G pro-tein antagonist, leads to the decrease of [Ca2+]c. Both L-type Ca2+channel blocker verapamil and inhibitor of IP3receptor heparin inhibitCTX-induced [Ca2+]cincrease. The results show that G protein may play a role in the modulation of [Ca2+]cthrough enhancing the ex-tracellular Ca2+influx and releasing of Ca2+from intracellular stores.

关键词： heterotrimeric G protein Lilium daviddi pollen cytoplasmic calcium.

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Differential display analysis of gene expression in Arabidopsis seedlings treated by Verticillium dahliae toxin

Differential display analysis of gene expression in Arabidop...

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2006年中国植物逆境生理生态与分子生物学学术研讨会

作者： Yan-Yan Hou Ying-Zhang Li (State Key laboratory of plant physiology and biochemistry, college of biological sciences, china agricultural university, beijing 100094

＜正＞Characterization of defense genes should permit greater understanding of defense response mechanisms at the molecular level and identify genes involved in defense response. In this aim we use the differential display technique to investigate the pattern of gene expression in Arabidopsis seedlings treated by Verticillium dahliae toxin （VD-toxin）. The cloning and the sequencing of two genes （DD1, DD2） isolated by this method are reported here. Difference in corresponding mRNA levels during 0-48 h of treatment was further confirmed by real-time RT-PCR.

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Functional Analysis of LTS1 Gene in plant Response to Low Temperature

Functional Analysis of LTS1 Gene in Plant Response to Low Te...

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2009整合植物生物学研讨会

作者： Haibian Yang and Shuhua Yang~* State Key laboratory of plant physiology and biochemistry,college of biological sciences,china agricultural university,beijing 100193,china.

Temperature is one of the most important environmental factors that affect plant growth and *** further understand the molecular mechanisms of plant response to low temperature,one mutant lts1-1(low temperature sensitive1)was *** normal growth condition(22℃),lts1-1 resembled the wild type ***,when grown at 4℃it

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The Crosstalk of Cold Stress and Defense Responses in Arabidopsis

The Crosstalk of Cold Stress and Defense Responses in Arabid...

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植物分子生物学与现代农业——2010全国植物生物学研讨会

作者： ShuhuaYang~*,Xiaozhen Huang,and Haibian Yang State Key laboratory of plant physiology and biochemistry,college of biological sciences,china agricultural university,beijing 100193,china

Low temperature is one of environmental factors that restrict plant growth homeostasis and plant-pathogen *** studies suggest a link between temperature responses and defense responses;however,the underlying molecular... 详细信息

关键词： low temperature defense responses chs mutants Arabidopsis

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