Journal of Plant Molecular Breeding

High-throughput direct regeneration of soybean mutant and common lines from cotyledonary node

Document Type : Original research paper

Authors

1 Department of Agronomy and Plant Breeding Sciences, College of Aburaihan, University of Tehran, Tehran-Pakdasht, Iran

2 Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology,Tehran, Iran

Abstract

Direct regenerations by using mature cotyledonary node as a explants has been shown to be time-saving and convenient strategy for micropropagation of soybean. So we have evaluated regeneration protocol through single shoot using cotyledonary node as a rapid and efficient protocol for two soybean cultivars and one mutant line. Cotyledonary nodes explants obtained from 7-days-old in vitro seedlings. After 28 days, the percent of regeneration and after 42 days, regeneration area were calculated. The results showed that percent of regeneration and regeneration area of mutant line was significantly more than two cultivars, L17 and Williams. After shoot induction, plants were transferred to shoot elongation medium followed by transferring plants to rooting medium. The results showed that the percent of rooting was not significantly affected by genotypes. In another experiment to test kanamycin sensitivity of regenerated shoots, it was found that kanamaycin with 150 mg/L concentration is lethal for regeneration of soybean shoots from cotyledonary node explants. The results showed that regeneration efficiency of mutant line was significantly more than two other cultivars. Kanamaycin sensitivity of regenerated shoots showed that kanamycin at 150 mg/L or above can be used as a selective agent for all three tested cultivars transformation.

Keywords

- References
[1]        Anita, R., Kuldeep, V. and Raman, S. 2012. Recovery of fertile transgenic plants via Agrobacterium tumefaciensmediated transformation in Indian soybean (Glycine max L. Merrill) cultivar. Indian J Genet Plant Breed, 72( 3): 325-331.
[2]        Arum, M., Subramanyam, k., Theboral, J. and Ganpathi, A. 2014. Optimized shoot regeneration for Indian soybean: the influence of exogenous polyamines. Plant Cell Tiss Organ Cult, 117(2): 305-309.
[3]        Barwale, U.B., Kerns, H.R. and Widholm, J.M. 1986. Plant regeneration from callus cultures of several soybean genotypes via embryogenesis and organogenesis. Planta, 167: 473-481.
[4]        Charbaji, T. and Nablusi, I. 1999. Effect of low doses of gamma irradiation on in vitro growth of grapevine. Plant Cell Tiss Organ Cult, 57:129-132.
[5]        Di, R., V. Purcell, G.B. Collins & S.A. Ghabrial, 1996. Production of transgenic soybean lines expressing the bean pod mottle virus coat protein precursor gene. Plant Cell Rep, 15: 746–750.
[6]        Fowlere, D.B. and Macqueen,  K.F. 1972. Effect of low doses of gamma radiation on yield and other agronomic characters of spring wheat (Triticum aestivum). Radiat. Bot, 12: 349-353.
[7]        Haliloglu, K. 2006. Efficient regeneration system from wheat leaf base segments . Biol Plantrum,50 (3): 326-330.
[8]        Hasbullah, N.A., Taha, R.M., Saleh, A. and Mahamad, N. 2012. Irradiation effect on in vitro organogenesis, callus growth and plantlet development of Gerbera jamesonii. Horti Bras, 30: 252-257.
[9]        Humera, A. 2006. Biochemical and Molecular Markers of Somaclonal Variants and Induced Mutants of Potato (Solanum tuberosum L.). An unpublished PhD dissertation.University of the Punjab Lahore, Pakistan.
[10]      Kang, E.J., Lee, Y.M., Sung, S.Y., Ha, B.K., Kim, S.H., Kim, D.S., Kim, J.B. and Kang, S.Y. 2013. Analysis of the Genetic Relationship of Gamma-irradiated in vitro mutants derived from standard-type Chrysanthemum cv. Migok. Hort Environ Biotechnol, 54(1):76-81.
[11]      Kita, Y., Nishizawa, K., Takahashi, M., Kitayama, M. and  Ishimoto, M. 2007. Genetic improvement and regeneration in soybean and transformation of the improved breeding lines. Plant Cell Rep, 26: 439-447.
[12]      Ma, X.H. and Wu, T.L. 2008. Rapid and efficient regeneration in soybean [Glycine max (L.) Merrill] from whole cotyledonary node explants. Acta Physiol Plant, 30:209–216.
[13]      Maghraby, G.M. 1987. Effect of radiation and some micronutrients on some legumes. Thesis submitted in partial fulfilment of the requirements for the degree of Master of Science. Zagazig University (Banha Branch).
[14]      Masuda, T. and Goldsmith, P.D. 2009. World soybean production: Area harvested, yield, and long-term projections. Int Food Agribusiness Manag Rev, 12: 143-162.
[15]      Mujar, E.K., Sidlik, N.J., Sulong, N.A., Jappar, S.S. and Othman, M.H. 2014. Effect of Low Gamma Radiation and Methyl Jasmonate on Vanilla planifolia Tissue Culture. Int J Pharm Sci Rev Res, 27(1): 163-167.
[16]      Olhoft, P.M., Flagel, L.E., Donovan, C.M. and Somers, D.A. 2003. Efficient soybean transformation using hygromycin B selection in the cotyledonary-node method. Planta, 216(5): 723-735.
[17]      Orazem, P., Stajner, N. and Bohanec, B. 2013. Effect of X-ray irradiation on olive shoot culture evaluated by morphological measurements, nuclear DNA content and SSR and AFLP markers. Trees, 27:1587–1595.
[18]      Patade, V.Y. and Suprasanna, P. 2008. Radiation induced in vitro mutagenesis for sugarcane improvement. Sugar Tech, 10(1): 14-19.
[19]      Paz, M.M., Martinez, J.C., Kalvig, A.B., Fonger, T.M., and Wang, K. 2006. Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. Plant Cell Rep, 25(3): 206-213.‏
[20]      Paz, M.M., Shou, H., Guo, Z., Zhang, Z., Banerjee, A. K., and Wang, K. 2004. Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant. Euphytica, 136(2): 167-179.‏
[21]      Yaycili, O. and Alikamanoglu, S. 2012. Induction of salt-tolerant potato (Solanum tuberosum L.) mutants with gamma irradiation and characterization of genetic variations via RAPD-PCR analysis. Turk J Biol, 36:405-412.
[22]      Younessi-Hamzekhanlu M., Izadi-Darbandi, A., Pirvali-Beiranvand, N., Halljian, M.T. and Majdabadi, A. 2011. Phenotypic and molecular analysis of M7 generation of soybean mutant lines through random amplified polymorphic DNA (RAPD) marker and some morphological traits. Afr J Agric Res, 6(7): 1779-1785.
 
 
Journal of Plant Molecular Breeding
Volume 3, Issue 1
June 2015
Pages 50-60
  • Receive Date: 26 January 2015
  • Revise Date: 26 August 2015
  • Accept Date: 26 August 2015
  • First Publish Date: 26 August 2015