质膜Ca2+-ATPase及H+-ATPase与细胞信号转导研究

批准号:
39970079
项目类别:
面上项目
资助金额:
12.0 万元
负责人:
王小菁
依托单位:
学科分类:
C0207.植物生殖与发育
结题年份:
2002
批准年份:
1999
项目状态:
已结题
项目参与者:
陈兆平、李德红、文彬、谢佐桂、郭丽荣
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中文摘要
在以往用原生质体实验系统,比较研究钙信使在光和激素信号转导中作用的基础上,分离纯化原生质膜,测定光和激素刺激后,质膜Ca2+-ATPase和H+-ATPase的变化,研究钙、钙调素的作用,并探讨光和激素刺激对酶的磷酸化、去磷酸化的影响。拟在同一实验体系比较光和激素信号转导的异同之处。此项课题对深入细致地研究植物信号转导的机理有重要理论意义。
英文摘要
Jasmonates (JAs) are "stress hormone" involved in signaling of defense responses. Our present knowledge of JAs signaling pathway is still limited. In the present study we investigated the effect of MeJA on H+-ATPase hydrolysis activity of the plasma membrane (PM ) isolated and purified from mung bean hypocotyls by aqueous two-phase partition and on the the phosphorylation and dephosphorylation of the enzyme comparing to fusicosin (FC) which is an activator of H+-ATPase. Moreover, the role of the calcium ion in MeJA- and FC-induced increases of the plasma membrane H+-ATPase was studied. .Application of MeJA (50μmol/L) to 3-d-old seedlings resulted in an increase (about 30%) in PM H+-ATPase activity in vivo. The enzyme activity after 10μmol/L MeJA treatment in vitro also showed about 30% increase 2hr after incubation. FC stimulated PM H+-ATPase activity and the maximal increase reached to 72% in vivo, while it had the same stimulation (about 30%) as MeJA had in vitro. The combination of MeJA and FC did not show significant additive effect on enzyme activity. In vitro, phosphatase inhibitors, okadaic acid and cantharidin, enhanced MeJA- induced increase of the enzyme activity. Staurosporine and cheleythrine, two inhibitors of protein kinase, inhibited the stimulation of MeJA on PM H+-ATPase activity completely. Both protein kinase and phosphatase inhibitors showed the same effect on FC-induced increases of enzyme activity. The results from γ-32p tracing experiments showed that the level of isotope labeling on PM H+-ATPase increased after treatment with MeJA and FC, respectively, demonstrating that the phosphorylation took place in the enzyme after MeJA treatment. Ca2+ strongly stimulated the PM H+-ATPase in vitro, and the increase of the enzyme activity was two times higher than that of the control. But Ca2+ had no enhancement of the enzyme activity induced by FC and MeJA .Trypsin digestion experiment was conducted to investigate the MeJA effect. After digesting and removing of a 7-10 kD segment of PM H+-ATPase C-terminus, the PM H+-ATPase activity increased by 30%. Same increasing of the enzyme activity was observed after MeJA and FC treatments, indicating the regulation of C-terminus of PM H+-ATPase might be involved in MeJA stimulation.in mung bean.[Ca2+]c changes were directly measured in Arabidopsis leaf from 10 d seedlings by using confocal laser scanning microscopy in conjunction with a calcium dye of Fluo-3AM. We found that [Ca2+]c increased rapidly after JA (50-150 mm) treatment. This increasing was inhibited by the pretreatment of the leaf with nifedipine, a Ca2+ channel blocker. But verlapamil, another Ca2+ channel blocker had no significant effect on JA induced [Ca2+]c changes. Moreover, the expression of two JA induced genes, vsp and jr1 were also inhibited when nifedipine was applied. And verlapamil could not affect two gene expression..Our data reported here demonstrated firstly that MeJA activated PM H+-ATPase hydrolysis activity and the the phosphorylation and dephosphorylation of the enzyme were involved in MeJA stimulation in mung bean; Ca2+/CaM were required in the JA signaling and nifedipine-sensitive channel was involved in [Ca2+]c changes induced by JA in Arabidopsis.
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DOI:10.1007/s10725-004-4454-6
发表时间:2004-11-01
期刊:PLANT GROWTH REGULATION
影响因子:4.2
作者:Meng, XC;Xing, T;Wang, XJ
通讯作者:Wang, XJ
DOI:--
发表时间:--
期刊:植物学通报
影响因子:--
作者:文彬;宾金华;潘瑞炽;王小菁
通讯作者:王小菁
DOI:--
发表时间:--
期刊:植物学通报
影响因子:--
作者:孙清鹏;王小菁
通讯作者:王小菁
Influx of extracellular Ca2 involved in jasmonic-acid-induced elevation of [Ca2 ]cyt and JR1
细胞外 Ca2 的流入参与茉莉酸诱导的 [Ca2 ]cyt 和 JR1 升高
DOI:--
发表时间:--
期刊:Journal of Plant Research
影响因子:2.8
作者:Qing-Peng Sun1;3;Yi Guo2;Ying Sun2;Da-Ye Sun2;Xiao-Jing Wang1;4
通讯作者:4
DOI:--
发表时间:--
期刊:热带亚热带植物学报
影响因子:--
作者:孙清鹏;王小菁
通讯作者:王小菁
GA与ABA拮抗调控非洲菊花瓣伸展的转录因子鉴定和功能分析
- 批准号:31372099
- 项目类别:面上项目
- 资助金额:90.0万元
- 批准年份:2013
- 负责人:王小菁
- 依托单位:
GASA6在GA、ABA以及糖信号互作网络中的功能研究
- 批准号:90917011
- 项目类别:重大研究计划
- 资助金额:60.0万元
- 批准年份:2009
- 负责人:王小菁
- 依托单位:
PRGL在植物生长发育中的功能分析
- 批准号:30570165
- 项目类别:面上项目
- 资助金额:35.0万元
- 批准年份:2005
- 负责人:王小菁
- 依托单位:
高粱野生型和har1突变体CRY1和CRY2基因的克隆和分析
- 批准号:30170558
- 项目类别:面上项目
- 资助金额:18.0万元
- 批准年份:2001
- 负责人:王小菁
- 依托单位:
钙信使系统在植物激素信号传递中的作用
- 批准号:39470361
- 项目类别:面上项目
- 资助金额:6.0万元
- 批准年份:1994
- 负责人:王小菁
- 依托单位:
钙在红光抑制茎细胞伸长中的作用
- 批准号:39170088
- 项目类别:面上项目
- 资助金额:3.0万元
- 批准年份:1991
- 负责人:王小菁
- 依托单位:
国内基金
海外基金
