Analisis ko-ekspresi gen-gen regulasi upstream dari gen Dehydrin di tanaman tebu (Saccharum officinarum L.) pada kondisi cekaman kekeringan
DOI:
https://doi.org/10.22302/iribb.jur.mp.v88i2.396Keywords:
expression analysis, transcription factors, DHN promoterAbstract
Sugarcane plantations in Indonesia have been expanded and shifted to the marginal land characterized by long drought period, therefore, an attempt has been initiated to generate drought tolerance varieties through genetic engineering. It could be conducted by inserting the gene that involve in plant adaptation response to drought stress such as dehydrin (DHN) into sugarcane genome. The promoter of sugarcane DHN gene was isolated and transformed into sugarcane in the previous research. This study aimed to demonstrate the functionality of sugarcane DHN promoter through expression analysis of DHN regulatory genes that play a role in response to drought stress. Expression analyses using RT-qPCR were also conducted on regulatory genes of sugarcane that inserted by Pr-1DHNSo construct treated with drought stress. The results showed that the expressions of SoMYB, SoWRKY, SoNAC, and SoDHN genes were escalated on sugarcane 16 days after stress treatment ranging from 353 to 4067 folds relatively to untreated samples in which SoNAC gene showed the highest expression. On the other hand, the analysis on transgenic sugarcane carrying DHNpromoter construct showed SoNAC and SoDREB expression increased after 72 hours under drought stress. The expression values of SoNAC in transgenic and non-transgenic plants under drought condition were 4.79 and 4.99, respectively. Meanwhile, the expression values of SoDREB in transgenic and non-transgenic plants under drought condition were 13.2 and 13.3, respectively. The results of these experiments showed that the promoter construct of Pr-1DHNSo was induced by drought stress treatments highlighting the regulation of several upstream genes of SoDHN.
Downloads
References
Ben Amar S, H Safi, M Ayadi, J Azaza, H Khoudi, K Masmoudi & F Brini (2013). Analysis of the promoter activity of a wheat dehydrin gene (DHN-5) under various stress conditions. AJCS 7(12), 1875-1883.
Baillo EH, RN Kimoto, Z Zhang & P Xu (2019). Transcription factors associated with abiotic and biotic stress tolerance and their potential for crops improvement. Genes 10(771), 1-23.
Bajpai S & R Chandra (2015). Effect of waterlogging stress on growth characteristic and SOD gene exspression in sugarcane. IJSRP 5(1), 1-8.
Chang S, J Puryear & J Cairney (1993). A simple and efficient method for isolating RNA from pine trees. J Plant Mol Biol Rep 11, 98-100.
Chen Y, L Huang, H Yan, X Zhang, B Xu & X Ma (2016). Cloning and characterization of an ABA-independent DREB transcription factor gene, HcDREB2 in Hemartria compressa. J Heredit 153(3), 1-7.
Farrell RE (2010). RNA Methodologies : Laboratory Guide for Isolation and Characterization. London, Elsevier.
Ghomati R, PNG Rao, K Chandran & A Shelvi (2015). Adaptive responses of sugarcane to waterlogging stress: an over view. Sugar Tech 17(4), 325-338.
Graca JP, FA Rodrigues, JRB Farias, MCN Oliveria, CBH Campo & SM Zingaretti (2010). Physiological parameters in sugarcane cultivars submitted to water deficit. J Plant Physiol 22(3), 189-197.
Hanin M, F Brini, C Ebel, Y Toda, S Takeda & K Masmoudi (2011). Plant dehydrins and stress tolerance. Versatile protein for complex mechanisms. J Plant Signal Behav 6(10), 1503-1509.
Iskandar HM, RE Casu, AT Fletcher, S Schmidt, JS Xu, DJ Maclean, JM Manners & GD Bonnett (2011). Identification of drought-response genes and a study of their expression during sucrose accumulation and water deficit in sugarcane culms. BMC Plant Biology 11(12), 1-14.
Iskandar HM, D Widyaningrum & Suhandono S (2014). Cloning and characterization of P5CS1 and P5CS2 genes from Saccharum officinarum L. under drought stress. J Trop Crop Scien 1(1), 23-30.
Lee SC, SH Kim & SR Kim (2013). Drought inducible OsDhn1 promoter is activated by OsDREB1A and OsDREB1D. J Plant Biol 56, 115-121.
Li C, CK Y-Ng & F Liu-Min (2015). MYB transcription factors, active players in abiotic stress signaling. Environ Exp Bot 114, 80-91.
Minarsih H, S Suhandono, AK Fuadi, T Kristianti, RA Putranto, D Sukmadjaya & Sustiprijatno (2020). Isolation and characterization of dehydrin promoter region from sugarcane (Saccharum officinarum L.). Menara Perkebunan 88(1), 16-28.
Mizoi J, K Shinozaki & K Yamaguchi-Shinozaki (2012). AP2/ERF family transcription factors in plant abiotic stress responses. BBA Gene Reg Mech 1819(2), 86-96.
Nakashima K, H Takasaki, J Mizoi, K Shinozaki & K Yamaguchi-Shinozaki (2012). NAC transcription factors in plant abiotic stress responses. BBA Gene Reg Mech 1819(2), 97-103.
Nuruzzaman M, AM Sharoni & S Kikuchi (2013). Roles of NAC transcription factors in the regulation of biotic and abiotic stress responses in plants. Front Microbiol 4(248).
Pfaffl MW. 2004. A-Z of Quantitative PCR. California, International University Line.
Ramiro AD, DMM Pasarin, MDA Barbosa, FD Santos, SGP Gomez, MSM Junior, E Lam & H Carrer (2016). Expression of arabidopsis bax inhibitor-1 in transgenic sugarcane confers drought tolerance. J Plant Biotech 14, 1826-1837.
Sade N, E Galkin E & M Moshelion (2015). Measuring Arabidopsis, tomato and barley leaf relative water content (RWC). J Bio-protocol 5(8), 1-4.
Sade N, A Gebremedhin & M Moshelion (2012). Risk-taking plants: anisohydric behavior as a stress-resistance trait. J Plant Signal Behav 7(7), 767-770.
Sade N, BJ Vinocur, A Diber, A Shatil, G Ronen, H Nissan, R Wallach, H Karchi & M Moshelion (2009). Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion?. J New Phytol 181(3), 651-661.
Santana AP, EJA Robledo, JAZ Briseno, JTA Sumuano, VMG Mendoza, FE Gil, LD Alcaraz, E Castario, MAK Lianes, FS Teyer & LCR Zapata. (2017). Transcriptional profiling of sugarcane leaves and root under progresive osmotic stress reveals a regulatedcoordination of gene expression in a spatiotemporal manner. J PLoS ONE 12(12), 1-25.
Shirvastava AK & S Shirvastava (2016). Diversity the germplasm of Saccharum species and related genera availabel for use in directed breeding programmes for sugarcane improvment. J Curren Scienc 111(3), 475-483.
Suhandono S, A Apriyanto & N Ihsan (2014). Isolation and characterization of three cassava elongation factor 1 alpha (MeEF1A) promoters. J PLoS ONE 9(1), 1-9.
Downloads
Submitted
Accepted
Published
How to Cite
Issue
Section
License
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
