AJMB  Vol.3 No.1 , January 2013
Identification and characterization drought tolerance of gene LEA-D11 soybean (glycine max L. Merr) based on PCR-sequencing
ABSTRACT
Drought is one of the most damaging abiotic stress. Different plants response differently to drought stress. Abiotic stresses such as drought induced diverse physicological and molecular responses in plants. These responses include changes in gene expression. One of drought tolerance gene is a gene encoding dehydrin which is belongs to the group II or D-11 LEA protein family. LEA-D11 gene produce dehydrin protein which has a role in stabilization of membrane structures and protection of macromolecules in the presence of drought. The aims of the study was to identify and to characterize the LEA-D11 gene in various soybean varieties. This research used seven varieties of soybean: Tanggamus, Nanti, Seulawah, Tidar (drought tolerant), Wilis and Burangrang (drought moderate) and Detam-1 (drought susceptible). DNA genome of those varieties was isolated using the methods from Doyle & Doyle [1]. DNA amplification was conducted using Polymerase Chain Reaction (PCR) with specific primers designed based on GmLEA-D11 gene sequence database from the NCBI. The DNA targets were sequenced using automatic sequencing machine, ABI 3130xl Genetic Analyzer, in Eijkman Institution. The result of this study showed that the sequences of Gm-LEA-D11 gene possessed by drought tolerance varieties (Tanggamus, Nanti, Seulawah and Tidar) and moderately tolerance (Wilis and Burangrang) were similar. However, the sequence of GmLEA-D11 gene detected in the drought susceptible variety Detam-1 was different from the two groups. Similarity between drought tolerance and moderately tolerance indicate that there is not only LEA-D11 gene responsible to drought tolerance but also others. The primer and sequences GmLEA-D11 gene can be used as molecular marker and capable of differentiating between drought susceptible and drought moderate to drought tolerant.

Cite this paper
Savitri, E. , Basuki, N. , Aini, N. and Arumingtyas, E. (2013) Identification and characterization drought tolerance of gene LEA-D11 soybean (glycine max L. Merr) based on PCR-sequencing. American Journal of Molecular Biology, 3, 32-37. doi: 10.4236/ajmb.2013.31004.
References
[1]   Doyle, J.J. and Doyle, J.L. (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19, 11-15.

[2]   Zhu, J.K., Hasegawa, P.M. and Bressan, R. (1997) Molecular aspect of osmotic stress in plant. Critical Reviews in Plant Sciences, 16, 253-277

[3]   Dure, L. (1993) Structural motif in LEA proteins. In: Close, T.J. and Bray, E.A., Eds., Reponse of Plants to Cellular Dehydration during Environmental Stress, American Society of Plant Physiologist, Rockville, 91-103.

[4]   Ingram, J. and Barterls, D. (1996) The molecular basis of dehydration tolerance in plants. Annual Review of Plant Physiology and Plant Molecular Biology, 47, 277-403. doi:10.1146/annurev.arplant.47.1.377

[5]   Thomashow, M.F. (1999) Plant cold acclimation: Freezing tolerance gene and regulatory mechanism. Annual Review of Plant Physiology and Plant Molecular Biology, 50, 571-599

[6]   Close, T.J. (1997) Dehydrin: A commonly in the response of plants to dehydration protein. Physiologia Plantarum, 100, 291-296. doi:10.1111/j.1399-3054.1997.tb04785.x

[7]   Dure, L., Crouch, M., Harada, J., Ho, T.H.D., Mundy, J. and Quatrano, R. (1989) Common amino acid sequence domains among the lea protein of higher plants. Plant Molecular Biology, 12, 475-486. doi:10.1007/BF00036962

[8]   Close, T.J. (1996) Dehydrin: Emergence of a biochemical role of a family of plant dehydration protein. Physiologia Plantarum, 97, 795-803. doi:10.1111/j.1399-3054.1996.tb00546.x

[9]   Bray, E.A. (1997) Plant responses to water deficit. Trends in Plant Science, 2, 48-54. doi:10.1016/S1360-1385(97)82562-9

[10]   Danyluk, J., Perron, A., Houde, M., Limin, A., Fowler, B., Benhamou, N. and Sarhan, F. (1998) Accumulation of an acidic dehydrin in the vicinity of the plasma membran during cold acclimation of wheat. The Plant Cell, 10, 623-638

[11]   Heyen, B.J., Alseikh, M.K., Smith, E.A., Torvik, C.F., Selas, D.F. and Randall, S.K. (2002) The calcium-binding activityn of vacuole associated, dehydin-like protein is regulated by phosporylation. Plant Physiology, 130, 675-687. doi:10.1104/pp.002550

[12]   Hara, M., Terashima, S., Fukaya, T. and Kuboi, T. (2003) Enhancement of cold tolerance and inhibition of lipid peroxidation by citrus dehydrin in transgenic tobacco. Planta, 217, 290-298

[13]   Shinozaki, K. and Yamaguchi-Shinozaki, K. (1996) Molecular responses to drought and cold stress. Current Opinion in Biotechnology, 7, 161-167. doi:10.1016/S0958-1669(96)80007-3

[14]   Imai, R., Chang, L., Ohta, A., Bray, E.A. and Takagi, M. (1996) A lea-class gene of tomato confers salt and freezing tolerance when overexpressing in Saccharomyces cerevisae. Gene, 170, 243-248. doi:10.1016/0378-1119(95)00868-3

[15]   Xu, D., Duan, X., Wang, B., Hong, B., Ho, T.H.D. and Wu, R. (1996) Expression of a late embryogenesis abundant protein gene, HVA 1 from barley confers tolerance to water deficits and salt stress in trangenic rice. Plant Physiology, 110, 249-257

[16]   Sivamani, E., Bahieldin, A., Wraith, J.M., Al-Niemo., T. Dyer, W.E., Ho, T.H.D. and Qu, R. (2000) Improved biomass productivity and water use efficiency under water deficit condition in transgenic wheat contituvely expressing the barley HVA 1 gene. Plant Science, 155, 1-9. doi:10.1016/S0168-9452(99)00247-2

[17]   Dure, L. (1993) A repeating 11-mer amino acid sequence domains among the LEA protein of higher plant. Plant Journal, 3, 363-369. doi:10.1046/j.1365-313X.1993.t01-19-00999.x

[18]   Hara, M., Fujinaga, M. and Kuboi, T. (2004) Radical scavenging activity and oxidative modification of citrus dehydrin. Plant Physiology and Biochemistry, 42, 657-662. doi:10.1016/j.plaphy.2004.06.004

[19]   Hara, M., Fujinaga, M. and Kuboi, T. (2005) Metal binding by citrus dehydrin with histidine-rich domains. Journal of Experimental Botany, 56, 2695-2703. doi:10.1093/jxb/eri262

[20]   Hara, M. (2009) The multifunctionality of dehydrins: An overview. Plant Signalling Behaviour, 5, 503-508

[21]   Gosal, S.S., Wani, S.H. and Manjit, S. (2009) Biotechnology and drought tolerance. Journal of Crop Improvement, 23, 19-54. doi:10.1080/15427520802418251

[22]   Novak, F.J. and Brunner, H. (1992) Plant breeding induced mutation technology for crop improvement. IAEA Bulleting, 4, 25-33.

[23]   Pahlevi, R. (2010) Study of marker specifity gene drought in variety of soybean (Glycine max) used PCR-sequencing. Thsesis, Post Graduate Programe, Brawijaya University, Malang.

[24]   Mahmudah (2009) Identification gene drought DREB1 and P5CS in varian soybean (Glycine max) from selection in vitro used method PCR-sequencing. Post Graduate Programe, Brawijaya University, Malang

[25]   Cellier, F., Conejero, G., Breitler, J.C. and Casse, F. (1998) Molecular and physological responses to water deficit in drought tolerant and drought sensitive lines in sunflower. Plant Physiology, 116, 319-328. doi:10.1104/pp.116.1.319

[26]   Choi, D.W., Zhu, B. and Close, T.J. (1990) The barley (Hordeum vulgare L.) dehydrin multigene family: Sequences, allele types, chromosome assignment, and expression characteristic of 11 Dhn genes of cv Dicktoo. Theoretical and Applied Genetics, 98, 1234-1247. doi:10.1007/s001220051189

[27]   Cohen, A. and Bray, E.A. (1992) Nocleotide sequence of an ABA-induced tomato genes that is expressed in wilted vegetative organs and developing seeds. Plant Molecular Biology, 18, 411-413. doi:10.1007/BF00034969

[28]   Godoy, J.A., Pardo, J.M. and Pintor-Toro, J.A. (1990) A tomato cDNA inducible by salt stress and absisic acid: Nucleotide sequence and expression pattern. Plant Molecular Biology, 15, 695-705. doi:10.1007/BF00016120

[29]   Giordani, T., Natali, L., D’Ercole, A., Pugliesi, C., Fambrini, M., Vernieri, P., Vitagliano, C. and Cavallini, A. (1999) Expression of a dehydrin gene during embryo development and drought stress in ABA-deficient mutants of sunflower (Helianthus annuus L.). Plant Molecular Biology, 39, 739-748. doi:10.1023/A:1006194720022

[30]   Shinozaki, K. and Yamaguchi-Shinozaki, K. (1997) Gene expression and signal transduction in water stress response. Plant Physiology, 115, 327-334. doi:10.1104/pp.115.2.327

[31]   Qiang, L., Zhao, N.M, Yamaguchi-Shinozaki, K. and Shinozaki, K (2000) Regulatory role od DREB transcription factors in plant drought, salt and cold tolerance. Chinese Science Bulletin, 45, 970-975. doi:10.1007/BF02884972

[32]   Porcel, R.B., dan Miguelm J. and Ruiz-Lozano, J.M. (2004) Evaluation of genes encoding for delta 1-pyroline-5-carboxylate synthase (P5CS) during drought stress in arbuscular mycorrhizal Glycine max and Lactuca sativa plants. http:www.ncbi.nlm.nih.gov

[33]   Robertson, M. and Chandler, P.M. (1992) Pea dehydrin: Identification, characterization and expression. Plant Molecular Biology, 19, 1031-1044. doi:10.1007/BF00040534

[34]   Colmenero-Flores, J.M., Campos, F. and Garciarrubias, A.A. (1997) Characterization of phaseolus vulgaris cDNA clones responsive to water deficit: Identification of a novel late embryogenesis abundant-like protein. Plant Molecular Biology, 35, 393-405. doi:10.1023/A:1005802505731

[35]   Zhang, J. and Kirkham, M.B. (2005) Enzymatic responses of the ascorbate-gluthatione cycle to drought in sorghum and sunflower plant. Plant Science, 113, 139-147. doi:10.1016/0168-9452(95)04295-4

[36]   Umezawa, T., Fujita., M., Fujita., Y., Yamaguchi-Shinozaki, K. and Shinozaki, K. (2006) Engineering drought tolerance in plants: Discovering and tailoring genes unlock the future. Current Opinion in Biotechnology, 17, 113-122. doi:10.1016/j.copbio.2006.02.002

 
 
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