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Licensed Unlicensed Requires Authentication Published by De Gruyter June 25, 2020

Overexpression of a novel E3 ubiquitin ligase gene from Coptis chinensis Franch enhances drought tolerance in transgenic tobacco

  • Hanting Chen , Junjun Li and Yang He EMAIL logo

Abstract

Drought stress has a significant effect on the growth, physiology and biochemistry of medicinal plants. SDIR1 (Salt- and Drought-Induced Ring Finger1), a C3H2C3-type RING-finger E3 ubiquitin ligase gene plays an important role in the stress response of various plants. However, the role of this gene is not clear in Coptis chinensis. In this study, the CcSDIR1 gene was cloned from C. chinensis using RACE and RT-PCR. Sequence analysis revealed that CcSDIR1 had an open reading frame of 840 bp that encodes 279 amino acids with a theoretical molecular weight about 31 kDa and pI value of 5.65 and shared conserved domains with other plants. On comparison with the wild-type plants, overexpression of CcSDIR1 in transgenic tobaccos increased drought tolerance and showed better growth performance. However, lower malondialdehyde contents and high antioxidant enzyme activities were observed in transgenic tobacco plants compared to wild-type plants. In addition, Evans blue staining showed high cell viability of transgenic lines under drought stress. These results suggest that CcSDIR1 regulates various responses to drought stress by increasing antioxidant enzyme activities and reducing oxidative damage. From the study results, the CcSDIR1 gene will be very useful for drought stress research in plants.


Corresponding author: Yang He, College of Medical Technology, State Key Laboratory of Characteristic Chinese Medicine Resources in Southwest China, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China, E-mail:

Funding source: Chengdu University of Traditional Chinese Medicine

Award Identifier / Grant number: CZYJC1904

Acknowledgment

This study was funded by project first-class disciplines development supported by Chengdu University of Traditional Chinese Medicine (CZYJC1904).

  1. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  2. Research funding: This study was funded by project first-class disciplines development supported by Chengdu University of Traditional Chinese Medicine (CZYJC1904).

  3. Conflict of interest: All authors declare that they have no conflict of interest.

References

1. Ramakrishna, A, Ravishankar, GA. Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav 2011;6:1720.10.4161/psb.6.11.17613Search in Google Scholar PubMed PubMed Central

2. Qi, S, Lin, Q, Zhu, H, Gao, F, Zhang, W, Hua, X. The RING finger E3 ligase SpRing is a positive regulator of salt stress signaling in salt-tolerant wild tomato species. Plant Cell Physiol 2016;57:528–39. http://doi.org/10.1093/pcp/pcw006.10.1093/pcp/pcw006Search in Google Scholar PubMed

3. Dreher, K, Callis, J. Ubiquitin, hormones and biotic stress in plants. Ann Bot 2007;99:787–822. http://doi.org/10.1093/aob/mcl255.10.1093/aob/mcl255Search in Google Scholar PubMed PubMed Central

4. Book, AJ, Smalle, J, Lee, KH, Yang, PZ, Walker, JM, Casper, S, et al. The RPN5 subunit of the 26S proteasome is essential for gametogenesis, sporophyte development, and complex assembly in Arabidopsis. Plant Cell 2009;21:460–78. http://doi.org/10.1105/tpc.108.064444.10.1105/tpc.108.064444Search in Google Scholar PubMed PubMed Central

5. Trujillo, M, Shirasu, K. Ubiquitination in plant immunity. Curr Opin Plant Biol 2010;13:402–8. http://doi.org/10.1016/j.pbi.2010.04.002.10.1016/j.pbi.2010.04.002Search in Google Scholar PubMed

6. Ryu, MY, Cho, SK, Hong, Y, Kim, J, Yang, SW. Classification of barley U-box E3 ligases and their expression patterns in response to drought and pathogen stresses. BMC Genomics 2019;20. https://doi.org/10.1186/s12864-019-5696-z.Search in Google Scholar

7. Stone, SL, Hauksdottir, H, Troy, A, Herschleb, J, Kraft, E, Callis, J. Functional analysis of the RING-type ubiquitin ligase family of Arabidopsis. Plant Physiol 2005;137:13–30. http://doi.org/10.1104/pp.104.052423.10.1104/pp.104.052423Search in Google Scholar PubMed PubMed Central

8. Ryu, MY, Cho, SK, Kim, WT. The Arabidopsis C3H2C3-type RING E3 ubiquitin ligase AtAIRP1 is a positive regulator of an abscisic acid-dependent response to drought stress. Plant Physiol 2010;154:1983–97. http://doi.org/10.1104/pp.110.164749.10.1104/pp.110.164749Search in Google Scholar PubMed PubMed Central

9. Ding, S, Zhang, B, Qin, F. Arabidopsis RZFP34/CHYR1, a ubiquitin E3 ligase, regulates stomatal movement and drought tolerance via SnRK2.6-mediated phosphorylation. Plant Cell 2015;27:3228–44. http://doi.org/10.1105/tpc.15.00321.10.1105/tpc.15.00321Search in Google Scholar PubMed PubMed Central

10. Zhang, Y, Yang, C, Li, Y, Zheng, N, Chen, H, Zhao, Q, et al. SDIR1 is a RING-finger E3 ligase that positively regulates stress-responsive abscisic acid signaling in Arabidopsis. Plant Cell 2007;19:1912–29. http://doi.org/10.1105/tpc.106.048488.10.1105/tpc.106.048488Search in Google Scholar PubMed PubMed Central

11. Zhang, YY, Li, Y, Gao, T, Zhu, H, Wang, DJ, Zhang, HW, et al. Arabidopsis SDIR1 enhances drought tolerance in crop plants. Biosci Biotechnol Biochem 2008;72:2251–4. http://doi.org/10.1271/bbb.80286.10.1271/bbb.80286Search in Google Scholar PubMed

12. Zhang, H, Cui, F, Wu, Y, Lou, L, Liu, L, Tian, M, et al. The RING finger ubiquitin E3 ligase SDIR1 targets SDIR1-INTERACTING PROTEIN1 for degradation to modulate the salt stress response and ABA signaling in Arabidopsis. Plant Cell 2015;27:214–27. http://doi.org/10.1105/tpc.114.134163.10.1105/tpc.114.134163Search in Google Scholar PubMed PubMed Central

13. Chen, HT, Fan, G, He, Y. Species evolution and quality evaluation of four Coptis herbal medicinal materials in Southwest China. 3 Biotech 2017;7:62. http://doi.org/10.1007/s13205-017-0679-8.10.1007/s13205-017-0679-8Search in Google Scholar PubMed PubMed Central

14. Chen, J, Wang, F, Liu, J, Lee, SC, Wang, X, Yang, H. Analysis of alkaloids in coptis chinensis Franch by accelerated solvent extraction combined with ultra performance liquid chromatographic analysis with photodiode array and tandem mass spectrometry detections. Anal Chim Acta 2008;613:184–95. http://doi.org/10.1016/j.aca.2008.02.060.10.1016/j.aca.2008.02.060Search in Google Scholar PubMed

15. Chen, HT, Deng, C, Nie, H, Fan, G, He, Y. Transcriptome analyses provide insights into the difference of alkaloids biosynthesis in the Chinese goldthread (Coptis chinensis Franch.) from different biotopes. PeerJ 2017;5:3303. http://doi.org/10.7717/peerj.3303.10.7717/peerj.3303Search in Google Scholar PubMed PubMed Central

16. Kong, W, Wei, J, Abidi, P, Lin, M, Inaba, S, Li, C, et al. Berberine is a novel cholesterol-lowering drug working through a unique mechanism distinct from statins. Nat Med 2004;10:1344–51. http://doi.org/10.1038/nm1135.10.1038/nm1135Search in Google Scholar PubMed

17. Choi, KB, Morishige, T, Shitan, N, Yazaki, K, Sato, F. Molecular cloning and characterization of coclaurinen-methyltransferase from cultured cells of Coptis japonica. J Biol Chem 2002;277:830–5. http://doi.org/10.1074/jbc.m106405200.10.1074/jbc.M106405200Search in Google Scholar PubMed

18. Ikezawa, N, Tanaka, M, Nagayoshi, M. Molecular cloning and characterization of CYP719, a methylened ioxy bridge-forming enzyme that belongs to a novel P450 Family from cultured Coptis japonica cells. J Biol Chem 2003;278:38557–65. http://doi.org/10.1074/jbc.m302470200.10.1074/jbc.M302470200Search in Google Scholar PubMed

19. He, Y, Hou, P, Fan, G, Li, D, Peng, C. Isolation and characterization of a novel (S)-canadine synthase gene from Coptis chinensis. Electron J Biotech 2015;18:376–80. http://doi.org/10.1016/j.ejbt.2015.08.004.10.1016/j.ejbt.2015.08.004Search in Google Scholar

20. Sato, F, Hashimoto, T, Hachiya, A, Tamura, K, Choi, KB, Morishige, T, et al. Metabolic engineering of plant alkaloid biosynthesis. P Natl Acad Sci USA 2001;98:367–72. http://doi.org/10.1073/pnas.98.1.367.10.1073/pnas.98.1.367Search in Google Scholar PubMed PubMed Central

21. Holsters, M, de Waele, D, Depicker, A, Messens, E. van Montagu, M, Schell, J. Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 1978;183:181–7. http://doi.org/10.1007/bf00267408.10.1007/BF00267408Search in Google Scholar PubMed

22. Horsch, RB, Fry, JE, Hoffmann, NL, Eichholtz, D, Rogers, SA, Fraley, RT. A simple and general method for transferring genes into plants. Science 1985;227:1229–31. http://doi.org/10.1126/science.227.4691.1229.10.1126/science.227.4691.1229Search in Google Scholar PubMed

23. Baker, CJ, Mock, NM. An improved method for monitoring cell death in cell suspension and leaf disc assays using evans blue. Plant Cell Tiss Org Cult 1994.39:7–12. https://doi.org/10.1007/BF00037585.Search in Google Scholar

24. Liu, N, Lin, ZF. Use of evans blue for testing cell viability of intact leaves of plant. Plant Physiol J 2011;47:570–4.Search in Google Scholar

25. Gouvea, DR, Gobbo-Neto, L, Lopes, NP. The influence of biotic and abiotic factors on the production of secondary metabolites in medicinal plants. Plant bioactives and drug discovery: principles, practice, and perspectives. John Wiley & Sons,Inc. 2012.10.1002/9781118260005.ch12Search in Google Scholar

26. Li, J, Fan, G, He, Y. Predicting the current and future distribution of three Coptis herbs in China under climate change conditions, using the MaxEnt model and chemical analysis. Sci Total Environ 2020;698:134141. http://doi.org/10.1016/j.scitotenv.2019.134141.10.1016/j.scitotenv.2019.134141Search in Google Scholar PubMed

27. Okada, N, KoIzumi, N, Tanaka, T, Ohkubo, H, Nakanishi, S, Yamada, Y. Isolation, sequence, and bacterial expression of a cDNA for (S)-tetrahydroberb erine oxidase from cultured berberine-producing Coptis japonica cells. P Natl Acad Sci USA 1989;86:534–8. http://doi.org/10.1073/pnas.86.2.534.10.1073/pnas.86.2.534Search in Google Scholar PubMed PubMed Central

28. He, Y, Hou, P, Fan, G, Arain, S, Peng, C. Comprehensive analyses of molecular phylogeny and main for Coptis (Ranunculaceae) species identification. Biochem Syst Ecol 2014;56:88–94. http://doi.org/10.1016/j.bse.2014.05.002.10.1016/j.bse.2014.05.002Search in Google Scholar

29. Tak, H, Mhatre, M. Molecular characterization of VvSDIR1 from Vitis vinifera and its functional analysis by heterologous expression in Nicotiana tabacum. Protoplasma 2013;250:565–76. http://doi.org/10.1007/s00709-012-0442-2.10.1007/s00709-012-0442-2Search in Google Scholar PubMed

30. Davey, MW, Stals, E, Panis, B, Keulemans, J, Swennen, RL. High-throughput determination of malondialdehyde in plant tissues. Anal Biochem 2005;347:201–7. http://doi.org/10.1016/j.ab.2005.09.041.10.1016/j.ab.2005.09.041Search in Google Scholar PubMed

31. Mittler, R, Vanderauwera, S, Gollery, M, Van Breusegem, F. Reactive oxygen gene network of plants. Trends Plant Sci 2004;9:490–8. http://doi.org/10.1016/j.tplants.2004.08.009.10.1016/j.tplants.2004.08.009Search in Google Scholar PubMed

32. Mallick, N, Mohn, FH. Reactive oxygen species: response of algal cells. J Plant Physiol 2000;157:183–93. http://doi.org/10.1016/s0176-1617(00)80189-3.10.1016/S0176-1617(00)80189-3Search in Google Scholar

33. Li, R, Wang, W, Wang, W, Li, F, Wang, Q, Xu, Y, et al. Overexpression of a cysteine proteinase inhibitor gene from Jatropha curcas confers enhanced tolerance to salinity stress. Electron J Biotech 2015;18:368–75. http://doi.org/10.1016/j.ejbt.2015.08.002.10.1016/j.ejbt.2015.08.002Search in Google Scholar

34. Lin, ZF, Liu, N, Lin, GZ, Peng, CL. Factors altering the membrane fluidity of spinach thylakoid as determined by fluorescence polarization. Acta Physiol Plant 2011;33:1019–24. http://doi.org/10.1007/s11738-011-0737-5.10.1007/s11738-011-0737-5Search in Google Scholar

Received: 2019-11-27
Accepted: 2020-05-31
Published Online: 2020-06-25
Published in Print: 2020-11-26

© 2020 Walter de Gruyter GmbH, Berlin/Boston

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