AJMB  Vol.2 No.1 , January 2012
Somaclonal variations of Soybeans (Glycine Max. L. Merr) stimulated by drought stress based on random amplified polymorphic DNAs (RAPDs)
In soybeans, drought stress causes 50% yield losses. Breeding for drought tolerance in soybeans has been widely developed using various methods, among which is polyethylene glycol (PEG-6000) induction to simulate drought in vitro. In a previous experiment, three somaclones with different levels of tolerance were generated. The objectives of this research were to determine the RAPD patterns of those somaclones and to investigate the correlation of the RAPD patterns to the drought tolerance characteristics. The results showed eleven RAPD primers capable of amplifying the DNA genome of soybeans, among which four primers were monomorphic and seven were polymorphic. Two of the polymorphic primers, OPK7 and OPK12, are capable of differentiating medium tolerance traits from other traits. Bands that are specific for medium tolerance against drought were 450 bp and 650 bp in size, generated by the OPK7 primer, and the band of 2000 bp, generated by the OPK12 primer. However, there was no band capable of differentiating between sensitive and tolerance varieties/lines, although some changing of the DNA sequence was detected in this research. This indicates that there are other factors responsible for the expression of drought tolerance.

Cite this paper
Arumingtyas, E. , Widoretno, W. and Indriyani, S. (2012) Somaclonal variations of Soybeans (Glycine Max. L. Merr) stimulated by drought stress based on random amplified polymorphic DNAs (RAPDs). American Journal of Molecular Biology, 2, 85-91. doi: 10.4236/ajmb.2012.21009.

[1]   Taji, T., Seki, M., Satou, M., Sakurai, T. Kobayashi, M., Ishiyama, K., Narusaka, Y., Narusaka, M., Zhu, J.K. and Shinozaki, K. (2004) Comparative genomics in salt tolerance between Arabidopsis and Arabidopsis-related halophyte salt cress using Arabidopsis microarray. Plant Physiology, 135, 1697-1709. doi:10.1104/pp.104.039909

[2]   Stolf-Moreira, R., Medri, M.E., Neumaier, N., Lemos, N.G., Pimenta, J.A., Tobita, S., Brogin, R.L., Marcelino-Guimar?es, F.C., Oliveira, M.C.N., Farias, J.R.B., Abdelnoor, R.V. and Nepomuceno, A.L. (2010) Soybean physiology and gene expression during drought. Genetics and Molecular Research, 9, 1946-1956. doi:10.4238/vol9-4gmr851

[3]   Widoretno, W., Guhardja, E., Ilyas, S. and Sudarsono. (2002) The effectivity of Polyetylene Glycol for evaluating the response of soybeans genotype toward drought stress in the seedling phase. Hayati, 9, 33-36.

[4]   Pastori, G.M. and Foyer, C.H. (2002) Common components networks and pathways of cross-tolerance to stress the central role of “redox” and abscisic acid-mediated. Plant Physiology, 129, 460-468. doi:10.1104/pp.011021

[5]   Shinozaki, K., Yamaguchi-Shinozaki, K. and Seki, M. (2003) Regulatory network of gene expression in the drought and cold stress responses. Current Opinion in Plant Biology, 6, 410-417. doi:10.1016/S1369-5266(03)00092-X

[6]   Bartels, D. and Sunkars, R. (2005) Drought and salt tolerance in plants. Critical Reviews in Plant Science, 24, 23-58. doi:10.1080/07352680590910410

[7]   Yamaguchi-Shinozaki, K. and Shinozaki, K. (2005) Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters. Trends in Plant Science, 10, 88-94. doi:10.1016/j.tplants.2004.12.012

[8]   Shinozaki, K. and Yamaguchi-Shinozaki, K. (2007) Gene networks involved in drought stress response and tolerance. Journal of Experimental Botany, 58, 221-22. doi:10.1093/jxb/erl164

[9]   Singh, K., Foley, R.C. and Onate-Sanchez, L. (2002). Transcription factors in plant defense and stress responses. Current Opinion in Plant Biology, 5, 430-436. doi:10.1016/S1369-5266(02)00289-3

[10]   Bocca, S.N., Magioli, C., Mangeon, A., Junqueira, R.M., Cardeal, V., Margis, R. and Sachetto-Martins, G. (2005). Survey of glycine-rich proteins (GRPs) in the Eucalyptus expressed sequence tag database (ForEST). Genetic and Molecular Biolology, 28, 608-624. doi:10.1590/S1415-47572005000400016

[11]   Hannah, M.A., Wiese, D., Freund, S., Fiehn, O., Heyer, A.G. and Hincha, D.K. (2006) Natural genetic variation of freezing tolerance in Arabidopsis. Plant Physiology, 142, 98-112. doi:10.1104/pp.106.081141

[12]   Aharon, R., Shahak, Y., Wininger, S., Bendov, R., Kapulnik, Y. and Galili, G. (2003) Overexpression of a plasma membrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but not under drought or salt stress. Plant Cell, 15, 439-447. doi:10.1105/tpc.009225

[13]   Aroca, R., Alguacil, M. M., Vernieri, P. and Ruiz-Lozano, M. (2008) Plant responses to drought stress and exogenous aba application are modulated differently by mycorrhization in tomato and an aba-desficient mutant (Sities). Microbial Ecology, 56, 704-719. doi:10.1007/s00248-008-9390-y

[14]   Schafleitner, R., Gaudin, A., Rosales, R.O.G., Aliaga, C.A.A. and Bonierbale, M. (2007) Proline accumulation and real time PCR expression analysis of genes encoding enzymes of proline metabolism in relation to drought tolerance in Andean potato. Acta Physiologiae Plantarum, 29, 19-26. doi:10.1007/s11738-006-0003-4

[15]   Iuchi, S., Kobayashi, M., Yamaguchi-Shinozaki, K. and Shinozaki, K. (2000) A stress-inducible gene for 9-cisepoxycarotenoid dioxygenase involved in abscisic acid biosynthesis under water stress in drought-tolerant cowpea. Plant Physiology, 123, 553-562. doi:10.1104/pp.123.2.553

[16]   Yamaguchi-Shinozaki, K. and Shinozaki, K. (2005) Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters. Trends in Plant Science, 10, 88-94. doi:10.1016/j.tplants.2004.12.012

[17]   Krishna, H. and Singh, S.K. (2007) Biotechnological advances in mango (Mangifera indica L.) and their future implication in crop improvement: A review. Biotechnology Advances, 25, 223-243. doi:10.1016/j.biotechadv.2007.01.001

[18]   Welsh, J. and McClelland, M. (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18, 7213-7218. doi:10.1093/nar/18.24.7213

[19]   Williams, J.G.K., Kubelik, A.K. and Livak, K.J. (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18, 6531-6535. doi:10.1093/nar/18.22.6531

[20]   Anping, G., Jianguang, S. and Xingi, C. (1998) Genome relationship among hibiscus section furcaria species based on random amplified polymorphic DNA (RAPD). Proceedings of the Workshop on Application of Biotechnology in the Improvement of Jute, Kenaf and Allied Fibres-Phase I, Beijing, China.

[21]   Cheng, Z., Lu, B. and Baldwin, B.S. (2002) Comparative studies of genetic diversity in kenaf (Hibiscus cannabinus L.) varieties based on analysis of agronomic and RAPD data. Hereditas, 136, 231-239. doi:10.1034/j.1601-5223.2002.1360309.x

[22]   Widoretno, W. and Sudarsono (2004) Evaluation on some lines of somaclonal variant soybeans arised from in vitro selection against drought. Hayati, 11, 11-20.

[23]   Widoretno, W., Guhardja, E., Ilyas, S. and Sudarsono (2003) Polyetilene glycol effectivity in evaluating response of soybeans to drought stress at germination stage. Hayati, 9, 33-36.

[24]   Trisnawati, L.P.A. (2006) Growth response, relative water content, and chlorophyll content after drought treatment of some soybean somaclones [Glycine Max (L)] resulted from in vitro selection using Polyetilene glycol at vegetative phase. Thesis, Faculty of Science, University of Brawijaya, Malang, Indonesia (unpublished).

[25]   Doyle J.J. and Doyle J.L. (1987) A rapid DNA isolation procedure from small quantities of fresh leaf tissues. Phytochemical Bulletin, 19, 11-15.

[26]   Balitkabi (Research Institute for Legumes and Tuber Crops) (2008) Description of superior legumes and tuber crops varieties. Research Institute for Legumes and Tuber Crops, Malang, Indonesia (unpublished).

[27]   Pakniyat, H. and Nazari L. (2008) Genetic diversity of wild and cultivated barley genotypes under drought stress using RAPD markers. Biotechnology Issue, 7, 745-750.

[28]   Li, Z. and Nelson, R.L. (2002) RAPD Marker Diversity among Cultivated and Wild Soybean Accessions from Four Chinese Provinces. Crop Science, 42, 1737-1744. doi:10.2135/cropsci2002.1737

[29]   Thompson, J.A. and Nelson, R.L. (1998) Core set of primers to evaluate genetic diversity in soybean. Crop Science, 38, 1356-1362. doi:10.2135/cropsci1998.0011183X003800050034x

[30]   Thompson, J.A., Nelson, R.L. and Vodkin, L.O. (1998) Identification soybean germplasm using RAPD markers. Crop Science, 38, 1348-1355. doi:10.2135/cropsci1998.0011183X003800050033x

[31]   Doldi, M.L., Vollmann, J. and Lelley, T. (2006) Genetic diversity in soybean as determined by RAPD and microsatellite analysis. Plant Breeding, 116, 331-335. doi:10.1111/j.1439-0523.1997.tb01007.x

[32]   Walters, C., Farrant, J.M., Pammenter, N.W. and Berjak, P. (2002) Desiccation stress and damage. In: Black, M. and Pritchard, H.W., Eds., Desiccation and Survival in Plants: Drying without Dying. CABI Publishing, Oxford and New York, 263-293. doi:10.1079/9780851995342.0263

[33]   Mundree, S.G., Baker, B., Mowla, S., Peters, S., Marais, S., Willigen, C.V., Govender, K, Maredza, A., Muyanga, S., Farrant, J.M. and Thomson J.A. (2002) Physiological and molecular insights into drought tolerance. African Journal of Biotechnology, 1, 28-38.

[34]   Verslues, P.E., Agarwal, M., Katiyar-Agarwal, S., Zhu, J. and Zhu, J.K. (2006) Techniques for Molecular Analysis: Methods and concepts in quantifying resistance to drought salt and freezing abiotic stresses that affect plant water status. The Plant Journal, 45, 523-539. doi:10.1111/j.1365-313X.2005.02593.x

[35]   Hamim, K., Ashari, Miftahudin and Triadiati (2008) Analysis of water status of prolin and the activity of antioxidant enzym on some drought tolerant, susceptible and wild type. Agrivita, 30, 201-210.

[36]   Simon-Sarkadi, L., Kocsy, G., Várhegyi, á., Galiba, G. and de Ronde, J.A. (2005) Genetic manipulation of proline accumulation influence the concentrations of other amino acids in soybean subjected to simultaneous drought and heat stress. Journal of Agricultural and Food Chemistry, 53, 7512-7517. doi:10.1021/jf050540l

[37]   Xiong, L., Schumaker, K.S. and Zhu, J.K. (2002) Cell Signaling during Cold Drought and Salt Stress. The Plant Cell, 14, 165-S183.

[38]   Hutami, S., Mariska, I. and Supriati, Y. (2006) The increase of genetic variability of plant through somaclonal variation. Jurnal AgroBiogen, 2, 81-88.

[39]   Chinnusamy, V., Schumaker, K, and Zhu, J.K. (2004). Molecular genetic perspectives on cross-talk and specificity in abiotic stress signalling in plants. Journal of Experimental Botany, 55, 225-236. doi:10.1093/jxb/erh005