DOI QR코드

DOI QR Code

The Arabidopsis Phytocystatin AtCYS5 Enhances Seed Germination and Seedling Growth under Heat Stress Conditions

  • Song, Chieun (Systems and Synthetic Agrobiotech Center and PMBBRC, Gyeongsang National University) ;
  • Kim, Taeyoon (Systems and Synthetic Agrobiotech Center and PMBBRC, Gyeongsang National University) ;
  • Chung, Woo Sik (Systems and Synthetic Agrobiotech Center and PMBBRC, Gyeongsang National University) ;
  • Lim, Chae Oh (Systems and Synthetic Agrobiotech Center and PMBBRC, Gyeongsang National University)
  • Received : 2017.05.12
  • Accepted : 2017.06.25
  • Published : 2017.08.31

Abstract

Phytocystatins (PhyCYSs) are plant-specific proteinaceous inhibitors that are implicated in protein turnover and stress responses. Here, we characterized a PhyCYS from Arabidopsis thaliana, which was designated AtCYS5. RT-qPCR analysis showed that the expression of AtCYS5 in germinating seeds was induced by heat stress (HS) and exogenous abscisic acid (ABA) treatment. Analysis of the expression of the ${\beta}-glucuronidase$ reporter gene under the control of the AtCYS5 promoter showed that AtCYS5 expression during seed germination was induced by HS and ABA. Constitutive overexpression of AtCYS5 driven by the cauliflower mosaic virus 35S promoter led to enhanced HS tolerance in transgenic Arabidopsis, which was characterized by higher fresh weight and root length compared to wild-type (WT) and knockout (cys5) plants grown under HS conditions. The HS tolerance of AtCYS5-overexpressing transgenic plants was associated with increased insensitivity to exogenous ABA during both seed germination and post-germination compared to WT and cys5. Although no HS elements were identified in the 5'-flanking region of AtCYS5, canonical ABA-responsive elements (ABREs) were detected. AtCYS5 was upregulated in ABAtreated protoplasts transiently co-expressing this gene and genes encoding bZIP ABRE-binding factors (ABFs and AREB3). In the absence of ABA, ABF1 and ABF3 directly bound to the ABREs in the AtCYS5 promoter, which activated the transcription of this gene in the presence of ABA. These results suggest that an ABA-dependent pathway plays a positive role in the HS-responsive expression of AtCYS5 during seed germination and post-germination growth.

Keywords

References

  1. Bae, E.K., Lee, H., Lee, J.S., and Noh, E.W. (2010). Isolation and characterization of osmotic stress-induced genes in poplar cells by suppression subtractive hybridization and cDNA microarray analysis. Plant Physiol. Biochem. 48, 136-141. https://doi.org/10.1016/j.plaphy.2009.11.002
  2. Chen, H., Hwang, J.E., Lim, C.J., Kim, D.Y., Lee, S.Y., and Lim, C.O. (2010). Arabidopsis DREB2C functions as a transcriptional activator of HsfA3 during the heat stress response. Biochem. Biophys. Res. Commun. 401, 238-244. https://doi.org/10.1016/j.bbrc.2010.09.038
  3. Christoff, A.P., Passaia, G., Salvati, C., Alves-Ferreira, M., Margis-Pinheiro, M., and Margis, R. (2016). Rice bifunctional phytocystatin is a dual molulator of legumain and papain-like proteases. Plant Mol. Biol. 92, 193-207. https://doi.org/10.1007/s11103-016-0504-5
  4. Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16, 735-743. https://doi.org/10.1046/j.1365-313x.1998.00343.x
  5. Fujita, Y., Fujita, M., Satoh, R., Maruyama, K., Parvez, M.M., Seki, M., Hiratsu, K., Ohme-Takagi, M., Shinozaki, K., and Yamaguchi-Shinozaki, K. (2005). AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis. Plant Cell 17, 3470-3488. https://doi.org/10.1105/tpc.105.035659
  6. Gabriele, S., Rizza, A., Martone, J., Circelli, P., Costantino, P., and Vittorioso, P. (2010). The Dof protein DAG1 mediates PIL5 activity on seed germination by negatively regulating GA biosynthetic gene AtGA3ox1. Plant J. 61, 312-323.
  7. Giuliano, G., Pichersky, E., Malik, V.S., Timko, M.P., Scolnik, P.A., and Cashmore, A.R. (1988). An evolutionary conserved protein binding sequence upstream of a plant light-regulated gene. Proc. Natl. Acad. Sci. USA 85, 7089-7093. https://doi.org/10.1073/pnas.85.19.7089
  8. Hong, J.K., Hwang, J.E., Lim, C.J., Yang, K.A., Jin, Z.L., Kim, C.Y., Koo, J.C., Chung, W.S., Lee, K.O., Lee, S.Y., et al. (2007). Over-expression of Chinese cabbage phytocystatin 1 retards seed germination in Arabidopsis. Plant Sci. 172, 556-563. https://doi.org/10.1016/j.plantsci.2006.11.005
  9. Huang, B., and Xu, C. (2008). Identification and characterization of proteins associated with plant tolerance to heat stress. J. Intergr. Plant Biol. 50, 1230-1237. https://doi.org/10.1111/j.1744-7909.2008.00735.x
  10. Hwang, J.E., Hong, J.K., Je, J., Lee, K.O., Kim, D.Y., Lee, S.Y., and Lim, C.O. (2009). Regulation of seed germination and seedling growth by an Arabidopsis phytocystatin isoform, AtCYS6. Plant Cell Rep. 28, 1623-1632. https://doi.org/10.1007/s00299-009-0762-7
  11. Hwang, J.E., Hong, J.K., Lim, C.J., Chen, H., Je. J., Yang, K.A., Kim, D.Y., Choi, Y.J., Lee, S.Y., and Lim, C.O. (2010). Distinct expression patterns of two Arabidopsis phytocystatin genes, AtCYS1 and AtCYS2, during development and abiotic stresses. Plant Cell Rep. 29, 905-915. https://doi.org/10.1007/s00299-010-0876-y
  12. Jakoby, M., Weisshaar, B., Dröge-Laser, W., Vicente-Carbajosa, J., Tiedemann, J., Kroi, T., and Parcy, F. (2002). bZIP transcription factors in Arabidopsis. Trends Plant Sci. 7, 106-111. https://doi.org/10.1016/S1360-1385(01)02223-3
  13. Je, J., Song, C., Hwang, J.E., Chung, W.S., and Lim, C.O. (2014) DREB2C acts as a transcriptional activator of the thermo tolerancerelated phytocystatin 4 (AtCYS4) gene. Transgenic Res. 23, 109-123. https://doi.org/10.1007/s11248-013-9735-2
  14. Jefferson, R.A., Kavanagh, T.A., and Bevan, M.W. (1987). GUS fusions: $\beta$-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901-3907.
  15. Julian, I., Gandullo, J., Santos-Silva, L.K., Diaz, I., and Martinez, M. (2013). Phylogenetically distant barley legumains have a role in both seed and vegetative tissues. J. Exp. Bot. 64, 2929-2941. https://doi.org/10.1093/jxb/ert132
  16. Kidric, M., Kos, J., and Sabotic, J. (2014). Proteases and their endogenous inhibitors in the plant to abiotic stress. Bot. Serb. 38, 139-158.
  17. Kim, S., Kang, J., Cho, D., Park, J.H., and Kim, S.Y. (2004). ABF2, an ABRE-binding bZIP factor, is an essential component of glucose signaling and its over expression affects multiple stress tolerance. Plant J. 40, 75-87. https://doi.org/10.1111/j.1365-313X.2004.02192.x
  18. Koo, J.C., Chun, H.J., Park, H.C., Kim, M.C., Koo, Y.D., Koo, S.C., Ok, H.M., Park, S.J., Lee, S.H., Yun, D.J., et al. (2002). Over-expression of a seed specific hevein-like antimicrobial peptide from Pharbitis nil enhances resistance to a fungal pathogen in transgenic tobacco plants. Plant Mol. Biol. 50, 441-452. https://doi.org/10.1023/A:1019864222515
  19. Lim, C.J., Hwang, .JE., Chen, H., Hong, J.K., Yang, K.A., Choi, M.S., Lee, K.O., Chung, W.S., Lee, S.Y., and Lim, C.O. (2007). Overexpression of the Arabidopsis DRE/CRT-binding transcription factor DREB2C enhances thermotolerance. Biochem. Biophys. Res. Commun. 362, 431-436. https://doi.org/10.1016/j.bbrc.2007.08.007
  20. Martinez, M., Rubio-Somoza, I., Fuentes, R., Lara, P., Carbonero, P., and Diaz, I. (2005). The barley cystatin gene (Icy) is regulated by DOF transcription factors in aleurone cells upon germination. J. Exp. Bot. 56, 547-556. https://doi.org/10.1093/jxb/eri033
  21. Park, H.C., Kim, M.L., Kang, Y.H., Jeon, J.M., Yoo, J.H., Kim, M.C., Park, C.Y., Jeong, J.C., Moon, B.C., Lee, J.H., et al., (2004). Pathogen- and NaCl-induced expression of the SCaM-4 promoter is mediated in part by a GT-1 box that interacts with a GT-1-like transcription factor. Plant Physiol. 135, 2150-2161. https://doi.org/10.1104/pp.104.041442
  22. Rodriguez, M.C., Edsgard, D., Hussain, S.S., Alquezar, D. Rasmussen, M., Gilbert, T., Nielsen, B.H., Bartels, D., and Mundy, J. (2010). Transcriptomes of the desiccation-tolerant resurrection plant Craterostigma plantagineum. Plant J. 63, 212-228. https://doi.org/10.1111/j.1365-313X.2010.04243.x
  23. Seki, M., Narusaka, M., Ishida, J., Nanjo, T., Fujida, M., Oono, Y., Kamiya, A., Nakajima, M., Enju, A., Sakurai, T., et al., (2002). Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J. 31, 279-292. https://doi.org/10.1046/j.1365-313X.2002.01359.x
  24. Song, C., Je, J., Hong, J.K., and Lim, C.O. (2014). Ectopic expression of an Arabidopsis dehydration-responsive element-binding factor DREB2C improves salt stress tolerance in crucifers. Plant Cell Rep. 33, 1239-1254. https://doi.org/10.1007/s00299-014-1612-9
  25. Song, C., Chung, W.S., and Lim, C.O. (2016). Overexpression of heat shock factor gene HsfA3 increases galactinol levels and oxidative stress tolerance in Arabidopsis. Mol. Cells 39, 477-483. https://doi.org/10.14348/molcells.2016.0027
  26. Sun, X., Yang, S., Sun, M., Wang, S., Ding, X., Zhu, D., Ji, W., Cai, H., Zhao, C., Wang, X., and Zhu, Y. (2014). A novel Glycine soja cysteine proteinase inhibitor GsCPI14, interacting with the calcium/calmodulinbinding receptor-like kinase GsCBRLK, regulated planttolerance to alkali stress. Plant Mol. Biol. 85, 33-48. https://doi.org/10.1007/s11103-013-0167-4
  27. Szewinska, J., Siminska, J., and Bielawski, W. (2016). The roles of cysteine proteases and phytocystatins in development and germination of cereal seeds. J. Plant Physiol. 207, 10-21. https://doi.org/10.1016/j.jplph.2016.09.008
  28. Tan, Y., Wei, X., Wang, P., Sun, X., Li, M., and Ma, F. (2016). A phytocystatin gene from Malus prunifolia (Willd.) Borkh., MpCYS5, confers salt stress tolerance and functions in endoplasmic reticulum stress response in Arabidopsis. Plant Mol. Biol. Rep. 34, 62-75. https://doi.org/10.1007/s11105-015-0900-z
  29. Toh, S., Imamura, A., Watanabe, A., Nakabayashi, K., Okamoto, M., Jikumaru, Y., Hanada, A., Aso, Y., Ishiyama, K., Tamura, N., et al. (2008). High temperature-induced abscisic acid biosynthesis and its role in the inhibition of gibberellins action in Arabidopsis seeds. Plant Physiol. 146, 1368-1385. https://doi.org/10.1104/pp.107.113738
  30. Vanderbeld, B., and Snedden, W.A. (2007). Developmental and stimulus-induced expression patterns of Arabidopsis calmodulin-like genes CML37, CML38 and CML39. Plant Mol. Biol. 64, 683-697. https://doi.org/10.1007/s11103-007-9189-0
  31. Van der Hoorn, R.A.L. (2008). Plant proteases: from phenotypes to molecular mechanisms. Annu. Rev. Plant Biol. 59, 191-223. https://doi.org/10.1146/annurev.arplant.59.032607.092835
  32. Verma, V., Ravindran, P., and Kumar, P. (2016). Plant hormonemediated regulation of stress responses. BMC Plant Biol. Doi 10.1186/s12870-016-0771-y.
  33. Zakharov, A., Carchilan, M., Stepurina, T., Rotari, V., Wilson, K., and Vaintraub, I. (2004). A comparative study of the role of the major proteinases of germinated common bean (Phaseolus vulgaris L.) and soybean (Glycine max (L.) Merrill) seeds in the degradation of their storage proteins. J. Exp. Bot. 55, 2241-2249. https://doi.org/10.1093/jxb/erh247
  34. Zhang, W., Ruan, J., Ho, T.D., You, Y., Yu, T., and Q, R.S. (2005). Cisregulatory element based targeted gene finding: genome-wide identification of abscisci acid- and abiotic stress-responsive genes in Arabidopsis thaliana. Bioinformatics 21, 3074-3081. https://doi.org/10.1093/bioinformatics/bti490
  35. Zhang, X., Liu, S., and Takano, T. (2008). Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance. Plant Mol. Biol. 68, 131-143. https://doi.org/10.1007/s11103-008-9357-x

Cited by

  1. VOZ1, a transcriptional repressor of DREB2C, mediates heat stress responses in Arabidopsis vol.247, pp.6, 2018, https://doi.org/10.1007/s00425-018-2879-9
  2. Impact of sirtuin-1 expression on H3K56 acetylation and oxidative stress: a double-blind randomized controlled trial with resveratrol supplementation vol.55, pp.4, 2018, https://doi.org/10.1007/s00592-017-1097-4
  3. A Highly Efficient Tumor-Targeting Nanoprobe with a Novel Cell Membrane Permeability Mechanism pp.09359648, 2019, https://doi.org/10.1002/adma.201807456
  4. Cellular NAD+Level: A Key Determinant of Mitochondrial Quality and Health vol.21, pp.4, 2017, https://doi.org/10.4235/agmr.2017.21.4.149
  5. Plant Cell Wall Proteomics: A Focus on Monocot Species, Brachypodium distachyon , Saccharum spp. and Oryza sativa vol.20, pp.8, 2017, https://doi.org/10.3390/ijms20081975
  6. Effect of supplementation of nicotinamide and sodium butyrate on the growth performance, liver mitochondrial function and gut microbiota of broilers at high stocking density vol.10, pp.11, 2017, https://doi.org/10.1039/c9fo00904c
  7. Inhibition of miR-383 suppresses oxidative stress and improves endothelial function by increasing sirtuin 1 vol.53, pp.2, 2017, https://doi.org/10.1590/1414-431x20198616
  8. Developmental Analysis of Mimulus Seed Transcriptomes Reveals Functional Gene Expression Clusters and Four Imprinted, Endosperm-Expressed Genes vol.11, pp.None, 2020, https://doi.org/10.3389/fpls.2020.00132
  9. Nicotinamide effects on the metabolism of human fibroblasts and keratinocytes assessed by quantitative, label-free fluorescence imaging vol.12, pp.10, 2021, https://doi.org/10.1364/boe.432561
  10. Updates on the Role of ABSCISIC ACID INSENSITIVE 5 (ABI5) and ABSCISIC ACID-RESPONSIVE ELEMENT BINDING FACTORs (ABFs) in ABA Signaling in Different Developmental Stages in Plants vol.10, pp.8, 2021, https://doi.org/10.3390/cells10081996
  11. Genome-Wide Identification and Characterization of the Cystatin Gene Family in Bread Wheat (Triticum aestivum L.) vol.22, pp.19, 2021, https://doi.org/10.3390/ijms221910264