Cloning and Expression Analysis of ThPP2C Gene from Tamarix hispida

doi:10.7525/j.issn.1673-5102.2017.03.010

Abstract

Abstract: Protein phosphatase 2C(PP2C) is a kind of serine/threonine protein phosphatase, which is present in the body as monomer. The reversible phosphorylation by PP2C catalytic is an important component of the cell signal transduction, which is involved almost all the physiological and pathological processes. A novel PP2C gene(ThPP2C) was isolated from Tamarix hispida by RT-PCR. The opening reading frame(ORF)length of ThPP2C was 869 bp, encoding 282 amino acid. The relative molecular weight of ThPP2C protein was 30.54 kD, with isoelectric point 7.13. By the real-time RT-PCR, the expressions of ThPP2C were obviously difference in roots and leaves of T.hispida under the treatments(0.4 mol·L^-1 NaCl, 20%(w/v) PEG₆₀₀₀, 150 μmol·L^-1 ABA or 100 μmol·L^-1 JA) compared to the control. The expression patterns of ThPP2C were not exactly similarity among these treatments. Particularly, the expression of ThPP2C in leaves of T.hispida after the treatments of NaCl, ABA and JA all were down-regulated. However, it was mainly up-regulated in roots under the treatments of NaCl and JA. With the treatment of ABA, the ThPP2C expression was down-regulated at the early stage and then gradually increased. Different with other three stress, under the treatment of PEG₆₀₀₀, the expressions of ThPP2C were obviously down-regulated in roots. It was not obviously different at the early stress stage in leaves until 48 h, when it was highly induced. Therefore, ThPP2C may play an important role in T.hispida to response the high salt, drought stress and hormones.

Key words: PP2C, Tamarix hispida, salt stress, ABA treatment

CLC Number:

WANG Pei-Long, LIU Zhong-Yuan, ZHANG Teng-Qian, TANG Fei-Fei, QU Guan-Zheng, GAO Cai-Qiu. Cloning and Expression Analysis of ThPP2C Gene from Tamarix hispida[J]. Bulletin of Botanical Research, 2017, 37(3): 395-401.

References

1. 周旺,亓晓艳,张应玖. 蛋白磷酸酶2C(PP2C)的表达、纯化与催化活性[J]. 吉林大学学报:理学版,2009,47(1):144-149.Zhou W,Qi X Y,Zhang Y J. Expression,purification and catalytic activity of protein phosphatase 2C(PP2C)[J]. Journal of Jilin University:Science Edition,2009,47(1):144-149.
2. Kharitonenkov A,Chen Z J,Sures I,et al. A family of proteins that inhibit signalling through tyrosine kinase receptors[J]. Nature,1997,386(6621):181-186.
3. Ruan H H,Yan Z H,Sun H,et al. The YCR079w gene confers a rapamycin-resistant function and encodes the sixth type 2C protein phosphatase in Saccharomyces cerevisiae[J]. FEMS Yeast Research,2007,7(2):209-215.
4. Schweighofer A,Hirt H,Meskiene I. Plant PP2C phosphatases:emerging functions in stress signaling[J]. Trends in Plant Science,2004,9(5):236-243.
5. Xue T T,Wang D,Zhang S Z,et al. Genome-wide and expression analysis of protein phosphatase 2C in rice and Arabidopsis[J]. BMC Genomics,2008,9:550.
6. Ma Y,Szostkiewicz I,Korte A,et al. Regulators of PP2C phosphatase activity function as abscisic acid sensors[J]. Science,2009,324(5930):1064-1068.
7. Sheen J. Mutational analysis of protein phosphatase 2C involved in abscisic acid signal transduction in higher plants[J]. Proceedings of the National Academy of Sciences of the United States of America,1998,95(3):975-980.
8. Nishimura N,Okamoto M,Narusaka M,et al. ABA Hypersensitive germination2-1 causes the activation of both abscisic acid and salicylic acid responses in Arabidopsis[J]. Plant and Cell Physiology,2009,50(12):2112-2122.
9. Hirayama T,Shinozaki K. Perception and transduction of abscisic acid signals:keys to the function of the versatile plant hormone ABA[J]. Trends in Plant Science,2007,12(8):343-351.
10. Jia H F,Lu D,Sun J H,et al. Type 2C protein phosphatase ABI1 is a negative regulator of strawberry fruit ripening[J]. Journal of Experimental Botany,2013,64(6):1677-1687.
11. Reyes D,Rodríguez D,González-garcía M P,et al. Overexpression of a protein phosphatase 2C from beech seeds in Arabidopsis shows phenotypes related to abscisic acid responses and gibberellin biosynthesis[J]. Plant Physiology,2006,141(4):1414-1424.
12. Saavedra X,Modrego A,Rodríguez D,et al. The nuclear interactor PYL8/RCAR3 of Fagus sylvatica FsPP2C1 is a positive regulator of abscisic acid signaling in seeds and stress[J]. Plant Physiology,2010,152(1):133-150.
13. Chérel I,Michard E,Platet N,et al. Physical and functional interaction of the Arabidopsis K⁺ channel AKT2 and phosphatase AtPP2CA[J]. The Plant Cell,2002,14(5):1133-1146.
14. Liu X,Zhu Y M,Zhai H,et al. AtPP2CG1,a protein phosphatase 2C,positively regulates salt tolerance of Arabidopsis in abscisic acid-dependent manner[J]. Biochemical and Biophysical Research Communications,2012,422(4):710-715.
15. Menges M,Hennig L,Gruissem W,et al. Cell cycle-regulated gene expression in Arabidopsis[J]. The Journal of Biological and Chemistry,2002,277(44):41987-42002.
16. 杜驰,张富春. 植物蛋白磷酸酶2C在非生物胁迫信号通路中的调控作用[J]. 生物技术通报,2014(8):16-22.Du C,Zhang F C. Protein phosphatases Ⅱ C in plants are involved in abiotic stress tolerance of several signaling pathways[J]. Biotechnology Bulletin,2014(8):16-22.
17. Merlot S,Gosti F,Guerrier D,et al. The ABI1 and ABI2 protein phosphatases 2C act in a negative feedback regulatory loop of the abscisic acid signalling pathway[J]. The Plant Journal,2001,25(3):295-303.
18. Yoshida T,Nishimura N,Kitahata N,et al. ABA-hypersensitive germination3 encodes a protein phosphatase 2C(AtPP2CA) that strongly regulates abscisic acid signaling during germination among Arabidopsis protein phosphatase 2Cs[J]. Plant Physiology,2006,140(1):115-126.
19. Tähtiharju S,Palva T. Antisense inhibition of protein phosphatase 2C accelerates cold acclimation in Arabidopsis thaliana[J]. The Plant Journal,2001,26(4):461-470.
20. Saez A,Apostolova N,Gonzalez-guzman M,et al. Gain-of-function and loss-of-function phenotypes of the protein phosphatase 2C HAB1 reveal its role as a negative regulator of abscisic acid signalling[J]. The Plant Journal,2004,37(3):354-369.

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