OVEREXPRESSION OF Gα GENE INCREASES GROWTH AND HYPOSALINE TOLERANCE IN Kappaphycus alvarezii TRANSGENIC PLANTLETS
Downloads
G proteins are membrane proteins that play roles in signal transduction in living organisms. They consist of α, β and γ subunits. The G protein α subunit (Gα) plays a role in plant resistance toward biotic and abiotic environmental stresses. Transgenic plantlets of Kappaphycus alvarezii carrying the Gα gene (derived from soybean) have been successfully obtained through Agrobacterium tumefaciens-mediated transformation. The present study aimed to: 1. compare the growth of non-transgenic and transgenic plantlets of K. alvarezii in vitro using Provasoli enriched seawater (PES) medium with normal salinity and hyposalinity and 2. analyze the expression level of the Gα gene in transgenic plantlets using quantitative Polymerase Chain Reaction (qPCR). The results showed that all transgenic plantlets (six clones) had significantly higher daily growth rate (DGR, % / d) than that of non-transgenic under the condition of normal salinity (30 ppt) and hyposalinity (15 and 20 ppt) for 5 weeks of observation. At 15 ppt, transgenic plantlets were more tolerant than non-transgenic ones, as most thalli of transgenic plantlets remained brown in color, whereas most thalli of non-transgenic plantlets were bleached. The results of the qPCR analysis showed that the expression of the Gα gene in transgenic plantlets increased by 6.43 - 8.03 times compared with that of non-transgenic plantlets. The result of Pearson correlation analysis showed that relative expression of Gα gene had a strong correlation, both with DGRs in normal salinity and hyposalinity of transgenic plantlets (correlation coefficient > 0.7). The correlation was linearly positive, where increased expression of the Gα gene was strongly associated with an increase in DGRs.
Aharon GS, Gelli A, Snedden WA, Blumwald E. 1998. Activation of a plant plasma membrane Ca²⁺ channel by T Gal, a heterotrimeric G protein α-subunit homologue. FEBS Lett 424:17–21.
Anderberg HI, Danielson J, Johanson U. 2011. Algal MIPS: high diversity and conserved motifs. BMC Evol Biol 11:110.
Badan Standarisasi Nasional. 2011. Cara Uji Kimia—Bagian 5: Penentuan Kadar Logam Berat Timbal (Pb) dan Kadmium (Cd) pada Produk Perikanan. SNI 2354.5:2011. Jakarta (ID): BSN.
Chakraborty N, Sharma P, Kanyuka K, Pathak RR, Choudhury D, Hooley R, Raghuram N. 2015. G protein α subunit (GPA1) regulates stress, nitrate and phosphate response, flavonoid biosynthesis, fruit/seed development and shares GCR1 regulation in Arabidopsis thaliana. Plant Mol Biol 89:513–525.
Chen JG, Gao Y, Jones AM. 2006. Differential roles of Arabidopsis heterotrimeric G-protein subunits in modulating cell division in roots. Plant Physiol 141:887–897.
Cheney DP, Metz B, Stiller J. 2001. Agrobacterium-mediated genetic transformation in the macroscopic marine red alga Porphyra yezoensis. J Phycol 37(Suppl.):11–12.
Colaneri AC, Tunc-Ozdemir M, Huang JP, Jones AM. 2014. Growth attenuation under saline stress is mediated by the heterotrimeric G protein complex. BMC Plant Biol 14:129.
Dawes CJ, Lluisma AO, Trono GC. 1994. Laboratory and field growth studies of commercial strains of Eucheuma denticulatum and Kappaphycus alvarezii in the Philippines. J Appl Phycol 6:21–24.
Fajri H. 2015. Genetic engineering of Nicotiana tabacum SR1 with Gα gene [thesis]. Bogor (ID): Institut Pertanian Bogor.
Gelli A, Higgins VJ, Blumwald E. 1997. Activation of plant plasma membrane Ca²⁺-permeable channels by race-specific fungal elicitors. Plant Physiol 113:269–279.
Gilliham M, Dayod M, Hocking BJ, Xu B, Conn SJ, Kaiser BN, Tyerman SD. 2011. Calcium delivery and storage in plant leaves: exploring the link with water flow. J Exp Bot 62(7):2233–2250.
Hasegawa PM. 2013. Sodium (Na⁺) homeostasis and salt tolerance of plants. Environ Exp Bot 92:19–31.
Hawkesford MJ, Horst W, Richey T, Lambers H, Schjoerring J, Møller IS, White P. 2012. Functions of macronutrients. In: Marschner P, editor. Marschner’s Mineral Nutrition of Higher Plants. 3rd ed. London (UK): Elsevier.
Hayashi L, Faria GSM, Nunes BG, Zitta CS, Scariot LA, Rover T, Felix MR, Bouzon ZL. 2010. Effects of salinity on growth rate, carrageenan yield, and cellular structure of Kappaphycus alvarezii cultured in vitro. J Appl Phycol 22:317–326.
Hepler PK. 2005. Calcium: a central regulator of plant growth and development. Plant Cell 17:2142–2155.
Huddy SM, Meyers AE, Coyne VE. 2012. Transformation of lacZ using different promoters in the commercially important red alga Gracilaria gracilis. Afr J Biotechnol 11:12321–12329.
Hurd CL, Harrison PJ, Bischof K, Lobban CS. 2014. Seaweed Ecology and Physiology. Cambridge (UK): Cambridge University Press.
Izawa Y, Takayanagi Y, Inaba N, Abe Y, Minami M, Fujisawa Y, Iwasaki Y. 2010. Function and expression pattern of the α subunit of the heterotrimeric G protein in rice. Plant Cell Physiol 51:271–281.
Karsten U. 2012. Seaweed acclimation to salinity and desiccation stress. In: Wiencke C, Bischof K, editors. Seaweed Biology. Berlin Heidelberg: Springer. p. 87–108.
Liu W, Ming Y, Li P, Huang Z. 2012. Inhibitory effects of hypo-osmotic stress on extracellular carbonic anhydrase and photosynthetic efficiency of Dunaliella salina. Plant Physiol Biochem 54:43–48.
Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2⁻ΔΔCT method. Methods 25:402–408.
Parenrengi A, Rachmansyah, Suryati E. 2011. Budidaya Rumput Laut Penghasil Karaginan (Karaginofit). Jakarta (ID): Badan Penelitian dan Pengembangan Kelautan dan Perikanan, KKP.
Page AF, Minocha SC. 2004. Analysis of gene expression in transgenic plants. In: Peña L, editor. Transgenic Plants: Methods and Protocols. Methods in Molecular Biology 286. p. 291–311.
Pei ZM, Gilroy S. 2008. Calcium signals and their regulation. Annu Rev Plant Biol 59:263–290.
Pratheesh PN, Shonima GM, Jiji Thomas, Abraham CJ, Muraleedhara Kurup G. 2012. Efficacy of different Agrobacterium tumefaciens strains in genetic transformation of Chlamydomonas reinhardtii. Adv Appl Sci Res 3(5):2679–2686.
Provasoli L. 1968. Media and prospects for the cultivation of marine algae. In: Watanabe A, Hattori A, editors. Cultures and Collections of Algae. Hakone (JP): Japanese Society of Plant Physiology.
Rajamuddin NIAL, Alimuddin, Widyastuti U, Faizal I. 2014. Evaluation of different promoters driving reporter gene expression in seaweed Kappaphycus alvarezii. Indones J Biotechnol 19(2):129–135.
Rajamuddin MAIA. 2016. Transformation of κ-carrageenase gene in seaweed Kappaphycus alvarezii [dissertation]. Bogor (ID): Institut Pertanian Bogor.
Rodriguez-Rosales MP, Gálvez FJ, Huertas R, Aranda MN, Baghour M, Cagnac O, Venema K. 2009. Plant NHX cation/proton antiporters. Plant Signal Behav 4:265–276.
Rumsey D. 2010. Statistics Essentials for Dummies. Indianapolis (US): Wiley Publishing.
Srinivasan R, Gothandam KM. 2016. Synergistic action of D-glucose and acetosyringone on Agrobacterium strains for efficient Dunaliella transformation. PLoS One 11:e0158322.
Sulistiani E, Yani SA. 2014. Kultur Jaringan Rumput Laut Kappaphycus alvarezii. Bogor (ID): SEAMEO BIOTROP.
Sulistiani S, Suharsono, Supena EDJ, Miftahudin. 2019. Agrobacterium-mediated genetic transformation of seaweed Kappaphycus alvarezii using Gα gene and callus cultures. BIOTROPIA 26(2):85–96.
Thuleau P, Schroeder JI, Ranjeva R. 1998. Recent advances in the regulation of plant calcium channels. Curr Opin Plant Biol 1:424–427.
Ullah H, Chen JG, Young JC, Im KH, Sussman MR, Jones AM. 2001. Modulation of cell proliferation by heterotrimeric G protein in Arabidopsis. Science 292:2066–2069.
Urano D, Chen JG, Botella JR, Jones AM. 2013. Heterotrimeric G protein signalling in the plant kingdom. Open Biol 3:120186.
Zhang W, Jeon BW, Assmann SM. 2011. Heterotrimeric G-protein regulation of ROS signalling and calcium currents in Arabidopsis guard cells. J Exp Bot 62(7):2371–2379.
Copyright (c) 2021 Erina Sulistiani
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Authors who publish with this journal agree with the following terms:
- Authors retain copyright and grant the journal right of first publication, with the work 1 year after publication simultaneously licensed under a Creative Commons attribution-noncommerical-noderivates 4.0 International License that allows others to share, copy and redistribute the work in any medium or format, but only where the use is for non-commercial purposes and an acknowledgement of the work's authorship and initial publication in this journal is mentioned.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
