拟南芥ABA受体与UGT71B6的相互作用研究
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摘要
为进一步分析ABA尿苷二磷酸葡萄糖基转移酶UGT71B6参与植物抗逆反应的具体机制,本研究通过体外酵母双杂交实验与GST-Pull down实验、体内双分子荧光互补实验(BIFC)分析了ABA受体RCARs与UGT71B6之间的相互作用关系.实验结果显示ABA受体RCAR12、RCAR13与UGT71B6,在体内、体外均存在直接的相互作用.研究表明UGT71B6的生理功能与ABA受体之间存在重要联系, UGT71B6可能通过ABA受体的介导参与了植物逆境应答反应.实验结果也为初步探究ABA受体是否参与ABA稳态调节过程提供一定参考.
In order to investigate whether ABA receptors participate in ABA homeostatic regulation mediated by UGT71 B6 and analyze the specific mechanisms of UGT71 B6's involvement in plant stress responses, we analyzed the interaction between RCARs and UGT71 B6 by yeast two-hybrid assay, BIFC, and GST-pull down assay. The results indicated that UGT71 B6 interacted with RCAR12/13 in vivo and in vitro. Thus, it is likely that the mechanism of UGT71 B6 participating in plant abiotic responses is related to its interaction with ABA RCARs. The experimental results also provide some references for the role studying of ABA receptors in the ABA homeostasis regulation process.
In order to investigate whether ABA receptors participate in ABA homeostatic regulation mediated by UGT71 B6 and analyze the specific mechanisms of UGT71 B6's involvement in plant stress responses, we analyzed the interaction between RCARs and UGT71 B6 by yeast two-hybrid assay, BIFC, and GST-pull down assay. The results indicated that UGT71 B6 interacted with RCAR12/13 in vivo and in vitro. Thus, it is likely that the mechanism of UGT71 B6 participating in plant abiotic responses is related to its interaction with ABA RCARs. The experimental results also provide some references for the role studying of ABA receptors in the ABA homeostasis regulation process.
引文
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[2] Wilkinson S, Davies W J. ABA-based chemical signalling: the co-ordination of responses to stress in plants [J]. Plant Cell Environ, 2002, 25: 195.
[3] Albinsky D, Masson J E, Bogucki A, et al. Plant responses to genotoxic stress are linked to an ABA/salinity signaling pathway [J]. Plant J, 1999, 17: 73.
[4] Wang P, Zhao Y, Li Z, et al. Reciprocal regulation of the TOR kinase and ABA receptor balances plant growth and stress response [J]. Mol Cell, 2018, 69: 100.
[5] Yamaguchi-Shinozaki K, Shinozaki K. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses [J]. Annu Rev Plant Biol, 2006, 57: 781.
[6] Xing L, Zhao Y, Gao J, et al. The ABA receptor PYL9 together with PYL8 plays an important role in regulating lateral root growth [J]. Sci Rep, 2016, 6: 27177.
[7] Lim C W, Lee S C. Arabidopsis abscisic acid receptors play an important role in disease resistance [J]. Plant Mol Biol, 2015, 88: 313.
[8] Aleman F, Yazaki J, Lee M, et al. An ABA-increased interaction of the PYL6 ABA receptor with MYC2 transcription factor: A putative link of ABA and JA signaling [J]. Sci Rep, 2016, 6: 28941.
[9] 李德款, 张亮, 彭梅芳, 等. ABA受体RCAR1/PYL9相互作用蛋白质的筛选及初步研究 [J]. 四川大学学报: 自然科学版, 2011, 48: 486.
[10] 刘震. 拟南芥尿苷二磷酸糖基转移酶(UGT71C5)对脱落酸(ABA)内稳态调节的分析 [D]. 成都: 四川大学, 2013.
[11] Qin X, Zeevaart J A. The 9-cis-epoxycarotenoid cleavage reaction is the key regulatory step of abscisic acid biosynthesis in water-stressed bean [J]. Proc Natl Acad Sci USA, 1999, 96: 15354.
[12] Lee K H, Piao H L, Kim H Y, et al. Activation of glucosidase via stress-induced polymerization rapidly increases active pools of abscisic acid [J]. Cell, 2006, 126: 1109.
[13] Xu Z Y, Lee K H, Dong T, et al. A vacuolar beta-glucosidase homolog that possesses glucose-conjugated abscisic acid hydrolyzing activity plays an important role in osmotic stress responses in Arabidopsis [J]. Plant Cell, 2012, 24: 2184.
[14] Okamoto M, Tanaka Y, Abrams S R, et al. High humidity induces abscisic acid 8′-hydroxylase in stomata and vasculature to regulate local and systemic abscisic acid responses in Arabidopsis [J]. Plant Physiol, 2009, 149: 825.
[15] Xu Z J, Nakajima M, Suzuki Y, et al. Cloning and characterization of the abscisic acid-specific glucosyltransferase gene from Adzuki bean seedlings. [J]. Plant Physiol, 2002, 129: 1285.
[16] 岑咏一, 刘震, 李旭锋, 等. UGT75B1、UGT71B6、UGT71C5 酶活性及酶反应动力学分析[J]. 四川大学学报: 自然科学版, 2016, 53: 171.
[17] Priest D M, Ambrose S J, Vaistij F E, et al. Use of the glucosyltransferase UGT71B6 to disturb abscisic acid homeostasis in Arabidopsis thaliana [J]. Plant J, 2006, 46: 492.
[18] Dong T, Xu Z Y, Park Y, et al. Abscisic acid uridine diphosphate glucosyltransferases play a crucial role in abscisic acid homeostasis in Arabidopsis [J]. Plant Physiol, 2014, 165: 277.
[2] Wilkinson S, Davies W J. ABA-based chemical signalling: the co-ordination of responses to stress in plants [J]. Plant Cell Environ, 2002, 25: 195.
[3] Albinsky D, Masson J E, Bogucki A, et al. Plant responses to genotoxic stress are linked to an ABA/salinity signaling pathway [J]. Plant J, 1999, 17: 73.
[4] Wang P, Zhao Y, Li Z, et al. Reciprocal regulation of the TOR kinase and ABA receptor balances plant growth and stress response [J]. Mol Cell, 2018, 69: 100.
[5] Yamaguchi-Shinozaki K, Shinozaki K. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses [J]. Annu Rev Plant Biol, 2006, 57: 781.
[6] Xing L, Zhao Y, Gao J, et al. The ABA receptor PYL9 together with PYL8 plays an important role in regulating lateral root growth [J]. Sci Rep, 2016, 6: 27177.
[7] Lim C W, Lee S C. Arabidopsis abscisic acid receptors play an important role in disease resistance [J]. Plant Mol Biol, 2015, 88: 313.
[8] Aleman F, Yazaki J, Lee M, et al. An ABA-increased interaction of the PYL6 ABA receptor with MYC2 transcription factor: A putative link of ABA and JA signaling [J]. Sci Rep, 2016, 6: 28941.
[9] 李德款, 张亮, 彭梅芳, 等. ABA受体RCAR1/PYL9相互作用蛋白质的筛选及初步研究 [J]. 四川大学学报: 自然科学版, 2011, 48: 486.
[10] 刘震. 拟南芥尿苷二磷酸糖基转移酶(UGT71C5)对脱落酸(ABA)内稳态调节的分析 [D]. 成都: 四川大学, 2013.
[11] Qin X, Zeevaart J A. The 9-cis-epoxycarotenoid cleavage reaction is the key regulatory step of abscisic acid biosynthesis in water-stressed bean [J]. Proc Natl Acad Sci USA, 1999, 96: 15354.
[12] Lee K H, Piao H L, Kim H Y, et al. Activation of glucosidase via stress-induced polymerization rapidly increases active pools of abscisic acid [J]. Cell, 2006, 126: 1109.
[13] Xu Z Y, Lee K H, Dong T, et al. A vacuolar beta-glucosidase homolog that possesses glucose-conjugated abscisic acid hydrolyzing activity plays an important role in osmotic stress responses in Arabidopsis [J]. Plant Cell, 2012, 24: 2184.
[14] Okamoto M, Tanaka Y, Abrams S R, et al. High humidity induces abscisic acid 8′-hydroxylase in stomata and vasculature to regulate local and systemic abscisic acid responses in Arabidopsis [J]. Plant Physiol, 2009, 149: 825.
[15] Xu Z J, Nakajima M, Suzuki Y, et al. Cloning and characterization of the abscisic acid-specific glucosyltransferase gene from Adzuki bean seedlings. [J]. Plant Physiol, 2002, 129: 1285.
[16] 岑咏一, 刘震, 李旭锋, 等. UGT75B1、UGT71B6、UGT71C5 酶活性及酶反应动力学分析[J]. 四川大学学报: 自然科学版, 2016, 53: 171.
[17] Priest D M, Ambrose S J, Vaistij F E, et al. Use of the glucosyltransferase UGT71B6 to disturb abscisic acid homeostasis in Arabidopsis thaliana [J]. Plant J, 2006, 46: 492.
[18] Dong T, Xu Z Y, Park Y, et al. Abscisic acid uridine diphosphate glucosyltransferases play a crucial role in abscisic acid homeostasis in Arabidopsis [J]. Plant Physiol, 2014, 165: 277.
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