Abstract
Ribosome-inactivating proteins (RIPs), a type of defensive chemicals, play key roles in host defense mechanisms against foreign pathogenic invaders and plant stress resistance. Alpha-momorcharin, a member of typical type I RIPs isolated from Momordica charantia, showed strongly antifungal, antitumor, and antibacterial activities both in vitro and in vivo. In this study, the expression level of alpha-momorcharin gene in response to environmental stress and phytohormones were examined through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting analyses. Results showed that α-momorcharin gene expression was up-regulated in response to salinity and osmotic stresses. Phytohormones, including abscisic acid and gibberellic acid, also enhanced α-momorcharin expression, while the expression level was not affected by salicylic acid. It’s speculated that the α-momorcharin gene might be functional in plant basal or defense resistance against environmental stresses, and it also involved in hormonal signaling pathways. This research might be exploited in further genetic engineering programs and plant protection.
Acknowledgements
This work was supported by research grants from the Doctor Fund of Huanggang Normal University (No. 2013030903), General Research Fund of Huanggang Normal University (No. 2014022203), and Fund of Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains (2015TD07).
References
Abebe T., Melmaiee K., Berg V. & Wise R.P. 2010. Drought response in the spikes of barley: gene expression in the lemma, palea, awn, and seed. Funct. Integr. Genomic 10: 191–205.10.1007/s10142-009-0149-4Search in Google Scholar PubMed
Apel K. & Hirt H. 2004. Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 55: 373–399.10.1146/annurev.arplant.55.031903.141701Search in Google Scholar PubMed
Au K.Y., Wang R.R., Wong Y.T., Wong K.B., Zheng Y.T. & Shaw P.C. 2014. Engineering a switch-on peptide to ricin A chain for increasing its specificity towards HIV-infected cells. BBA-Gen. Subjects 1840: 958–963.10.1016/j.bbagen.2013.11.005Search in Google Scholar PubMed
Böttger S., Westhof E., Siems K. & Melzig M.F. 2013. Structure-activity relationships of saponins enhancing the cytotoxicity of ribosome-inactivating proteins type I (RIP-I). Toxicon 73: 144–150.10.1016/j.toxicon.2013.07.011Search in Google Scholar PubMed
Chinnusamy V., Zhu J. & Zhu J. 2007. Cold stress regulation of gene expression in plants. Trends Plant Sci. 12: 444–451.10.1016/j.tplants.2007.07.002Search in Google Scholar PubMed
Corrado G., Scarpetta M., Alioto D., Di Maro A., Polito L., Parente A. & Rao R. 2008. Inducible antiviral activity and rapid production of the ribosome-inactivating protein I from Phytolacca heterotepala in tobacco. Plant Sci. 174: 467–474.10.1016/j.plantsci.2008.01.009Search in Google Scholar
de Virgilio M., Lombardi A., Caliandro R. & Fabbrini M.S. 2010. Ribosome-Inactivating Proteins: from plant defense to tumor attack. Toxins 2: 2699–2737.10.3390/toxins2112699Search in Google Scholar PubMed PubMed Central
Ganeshan S., Vitamvas P., Fowler D.B. & Chibbar R.N. 2008. Quantitative expression analysis of selected COR genes reveals their differential expression in leaf and crown tissues of wheat (Triticum aestivum L.) during an extended low temperature acclimation regimen. J. Exp. Bot. 59: 2393–2402.10.1093/jxb/ern112Search in Google Scholar PubMed PubMed Central
Garson J.A., Grant P.R., Ayliffe U., Ferns R.B. & Tedder R.S. 2005. Real-time PCR quantitation of hepatitis B virus DNA using automated sample preparation and murine cytomegalovirus internal control. J. Virol. Methods. 126: 207–213.10.1016/j.jviromet.2005.03.001Search in Google Scholar PubMed
Huang D., Wu W., Abrams S.R. & Cutler A.J. 2008. The relationship of drought-related gene expression in Arabidopsis thaliana to hormonal and environmental factors. J. Exp. Bot. 59: 2991–3007.10.1093/jxb/ern155Search in Google Scholar PubMed PubMed Central
Ishizaki T, Megumi C, Komai F, Masuda K. & Oosawa K. 2002. Accumulation of a 31-kDa glycoprotein in association with the expression of embryogenic potential by spinach callus in culture. Physiol. Plant. 114: 109–115.10.1034/j.1399-3054.2002.1140115.xSearch in Google Scholar PubMed
Lovdal T. & Lillo C. 2009. Reference gene selection for quantitative real-time PCR normalization in tomato subjected to nitrogen, cold, and light stress. Anal. Biochem. 387: 238–242.10.1016/j.ab.2009.01.024Search in Google Scholar PubMed
Mahajan S. & Tuteja N. 2005. Cold, salinity and drought stresses: an overview. Arch. Biochem. Biophys. 444: 139–158.10.1016/j.abb.2005.10.018Search in Google Scholar PubMed
Meloni D.A. & Martínez C.A. 2009. Glycinebetaine improves salt tolerance in vinal (Prosopis ruscifolia Griesbach) seedlings. Brazilian J. Plant Physiol. 21: 233–241.10.1590/S1677-04202009000300007Search in Google Scholar
Ostermaier V. & Kurmayer R. 2009. Distribution and abundance of nontoxic mutants of cyanobacteria in lakes of the Alps. Microbial Ecol. 58: 323–333.10.1007/s00248-009-9484-1Search in Google Scholar PubMed PubMed Central
Qin W., Ming-Xing H., Ying X., Xin-Shen Z. & Fang, C. 2005. Expression of a ribosome inactivating protein (curcin 2) in Jatropha curcas is induced by stress. J. Bioscience 30: 351–357.10.1007/BF02703672Search in Google Scholar PubMed
Reinbothe S., Reinbothe C., Lehmann J., Becker W., Apel K. & Parthier B. 1994. JIP60, a methy jasmonate-induced ribosome inactivating protein involved in plant stress reactions. Proc. Nat. Acad. Sci. USA 91: 7012–7016.10.1073/pnas.91.15.7012Search in Google Scholar PubMed PubMed Central
Stirpe F. 2013. Ribosome-inactivating proteins: From toxins to useful proteins. Toxicon 67: 12–16.10.1016/j.toxicon.2013.02.005Search in Google Scholar PubMed
Song S.K., Choi Y., Moon Y.H., Kim S.G., Choi Y.D. & Lee J.S. 2000. Systemic induction of a Phytolacca insularis antiviral protein gene by mechanical wounding, jasmonic acid, and abscisic acid. Plant Mol. Biol. 43: 439–450.10.1023/A:1006444322626Search in Google Scholar
Tarantini A., Pittaluga E., Marcozzi G., Testone G., Rodrigues-Pousada R.A., Giannino D. & Spanò L. 2010. Differential expression of saporin genes upon wounding, ABA treatment and leaf development. Physiol. Plant. 140: 141–152.10.1111/j.1399-3054.2010.01388.xSearch in Google Scholar PubMed
Umezawa T., Fujita M., Fujita Y., Yamaguchi-Shinozaki K. & Shinozaki K. 2006. Engineering drought tolerance in plants: discovering and tailoring genes to unlock the future. Curr. Opin. Biotech. 17: 113–122.10.1016/j.copbio.2006.02.002Search in Google Scholar PubMed
Vivanco J.M., Savary B.J. & Flores H.E. 1999. Characterization of two novel type I ribosome-inactivating proteins from the storage roots of the Andean crop Mirabilis expansa .Plant Physiol. 119: 1447–1456.10.1104/pp.119.4.1447Search in Google Scholar PubMed PubMed Central
Wan H., Zhao Z., Qian C., Sui Y., Malik A.A. & Chen J. 2010. Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber. Anal. Biochem. 399: 257–261.10.1016/j.ab.2009.12.008Search in Google Scholar PubMed
Wong R.N.S., Mak N.K., Choi W.T. & Law P.T.W. 1995. Increased accumulation of trichosanthin in Trichosanthes kirilowii induced by microorganisms. J. Exp. Bot. 46: 355–358.10.1093/jxb/46.3.355Search in Google Scholar
Xu J., Wang H. & Fan J. 2007. Expression of a ribosome-inactivating protein gene in bitter melon is induced by Sphaerotheca fuliginea and abiotic stimuli. Biotechnol. Lett. 29: 1605–1610.10.1007/s10529-007-9433-3Search in Google Scholar PubMed
©2016 Institute of Botany, Slovak Academy of Sciences
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Articles in the same Issue
- Cellular and Molecular Biology
- Antimicrobial resistance and molecular characterisation of human campylobacters from Slovakia
- Cellular and Molecular Biology
- Application of violet pigment from Chromobacterium violaceum UTM5 in textile dyeing
- Cellular and Molecular Biology
- Chemical composition and antioxidant activity of some important betel vine landraces
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- Botany
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- Botany
- Environmental factors and phytohormones enhancing expression of α-momorcharin gene in Momordica charantia
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