Biologia

, Volume 65, Issue 3, pp 496–503 | Cite as

Multiple shoots induction in wild cotton (Gossypium bickii) through organogenesis and the analysis of genetic homogeneity of the regenerated plants

  • Xi-Yan Yang
  • Xian-Long Zhang
  • Li-Li Fu
  • Ling Min
  • Guan-ze Liu
Section Botany

Abstract

Fertile plants were regenerated via organogenesis from cultures of Gossypium bickii Prokhanov cotyledonary nodes devoid of cotyledons. Cotyledonary nodes from 7-day-old seedlings yielded the maximum number of shoots (9.12 shoots per explant) using MSB medium [MS medium (Murashige & Skoog 1962) and B5 (Gamborg et al. 1968) vitamins] supplemented with the combination of 4 mg/L BAP (N6-benzyladinine) and 0.1 mg/L TDZ (thidiazouron), and the subculture was made on a medium containing 2 mg/L BAP. Elongation of multiple shoots was obtained on MSB medium containing 0.05 mg/L GA3 (gibberellic acid). For rooting, a modified half-strength MS medium was utilized. Some regenerants were transferred to a greenhouse, and most of them were morphologically normal and fertile. Histological studies revealed that proliferating buds originated directly from the superficial layers of the explants. Flow cytometric analyses and chromosome counting suggested that the regenerants and their offsprings were diploid. DNA content of regenerants and their offsprings was homogeneous and similar to that of seed-derived plants with unchanged chromosome number (2n = 26). Random amplified polymorphic DNA (RAPD) analysis confirmed the genetic homogeneity of the regenerants and their offsprings with the diploid parent.

Key words

Gossypium bickii Prokhanov BAP cotton genetic homogeneity multiple shoots TDZ 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Agrawal D.C., Banerjee A.K., Kolalal R.R., Dhage A.B., Kulkarni W. V. & Nalawade S.M. 1997. In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.). Plant Cell Rep. 16: 647–652.CrossRefGoogle Scholar
  2. Caboni E., Lauri P. & D’Angeli S. 2000. In vitro plant regeneration from callus of shoot apices in apple shoot culture. Plant Cell Rep. 19: 755–760.CrossRefGoogle Scholar
  3. Capelle S.C., Mok D.W.S. & Turner J.E. 1983. Effects of thidiazuron on cytokinin autonomy and the metabolism of N6-(D2-isopentenyl) [8–14C] adenosine in callus cultures of Phaseolus lunatus L.. Plant. Physiol. 73: 796–802.CrossRefPubMedGoogle Scholar
  4. Charisse M.B., Randy C.S. & Robert M.B. 1994. Early events of multiple bud formation and shoot development in Soybean embryonic axes treated with the cytokinin, 6-Benzylaminopurine. Am. J. Bot. 81(11): 1435–1448.CrossRefGoogle Scholar
  5. Finer J.J. & Smith R.H. 1984. Initiation of callus and somatic embryos from explants of mature cotton (Gossypium klotzschianum Anderess). Plant Cell Rep. 3: 41–43.CrossRefGoogle Scholar
  6. Firoozabady E. & DeBoer D.L. 1993. Plant regeneration via somatic embryogenesis in many cultivars of cotton (Gossypium hirstum L.). In vitro Cell Dev Biol. Plant. 29: 166–173.CrossRefGoogle Scholar
  7. Fryxell P.A. 1992. A revised taxonomic interpretation of Gossypium L. (Malvaceae). Rhoedea. 2: 108–165.Google Scholar
  8. Gamborg O.L., Miller R.A. & Ojima K. 1968. Nutrient requirements of suspension culture of soybean roots cells. Exp. Cell. Res. 50: 150–158.CrossRefGoogle Scholar
  9. Hazra S., Kulkarni A.V., Nalawade S.M., Banerjee A.K., Agrawal D.C. & Krishtamurthy K.V. 2000. Influence of explants, genotypes and culture vessels on sprouting and proliferation of pre-existing meristems of cotton (Gossypium hirsutum L.). In vitro Cell Dev. Biol. Plant. 36: 505–510.CrossRefGoogle Scholar
  10. Jain S.M. 2001. Tissue culture-derived variation in crop improvement. Euphytica. 118: 153–166.CrossRefGoogle Scholar
  11. Jin S.X., Zhang X.L., Liang S.G., Nie Y.C., Guo X.P. & Huang C. 2005. Factors affecting transformation efficiency of embryogenic callus of upland cotton (Gossypium hirsutum) with Agrobacterium tumefaciens. Plant Cell Tissue Organ Cult.81: 229–237.CrossRefGoogle Scholar
  12. Kumria R., Sunnichan V.G., Das D.K., Gupta S.K., Reddy V.S. & Bhatnagar, R.K. 2003. High-frequency somatic embryo production and maturation into normal plants in cotton (Gossypium hirsutum) through metabolic stress. Plant Cell Rep. 21: 635–639.PubMedGoogle Scholar
  13. Lacroix B., Assoumou Y. & Sangwan R.S. 2003. Efficient in vitro direct shoot organogenesis and regeneration of fertile plants from embryo explants of Bambara groundnut (Vigna subterranea L. Verdc.) Plant Cell Rep. 21: 1153–1158.CrossRefPubMedGoogle Scholar
  14. Lambe P., Mutambel H.S.N., Fouche J.G., Deltour R., Foidart J.M. & Gaspar T. 1997. DNA methylation as a key process in regulation of organogenic totipotency and plant neoplastic progression. In Vitro Cell Dev. Biol. Plant. 33: 155–162.CrossRefGoogle Scholar
  15. Lee S.H., Shon Y.G., Kim C.Y., Chun H.J., Cheong Y.H. & Kim Z.H. 1999. Variations in the morphology of rice plants regenerated from protoplasts using different culture procedures. Plant Cell Tiss Org Cult. 57: 179–187.CrossRefGoogle Scholar
  16. McClean R. & Grafton K.F. 1989. Regeneration of dry bean (Phaseolus vulgaris L.) via organogenesis. Plant Sci. 60: 117–122.CrossRefGoogle Scholar
  17. McStewart J.D. & Hsu C.L. 1977. In ovulo embryo culture and seedling development of cotton (Gossypium hirsutum L.). Planta. 137: 113–117.CrossRefGoogle Scholar
  18. Matt A. & Jehle J.A. 2005. In vitro plant regeneration from leaves and internode sections of sweet cherry cultivars (Prunus avium L.). Plant Cell Rep. 24: 468–476.CrossRefPubMedGoogle Scholar
  19. Murch S.J., Victor J.M.R., Krishnaraj S. & Saxena P.K. 1999. The role of proline in thidiazuron-induced somatic embryogenesis of peanut. In Vitro Cell Dev. Biol. Plant. 35: 102–105.Google Scholar
  20. Murthy B.N.S., Murch S.J. & Saxena P.K. 1995. Thidiazuron induced somatic embryogenesis in intact seedlings of peanut (Arachishypogaea L.): endogenous growth regulator levels and significance of cotyledons. Physiol. Plant. 94: 268–276.CrossRefGoogle Scholar
  21. Murthy B.N.S, Murch S.J. & Saxena P.K. 1998. Thidiazuron: a potent regulator of in vitro plant morphogenesis. In Vitro Cell Dev. Biol. Plant. 34: 267–275.CrossRefGoogle Scholar
  22. Murashige T. & Skoog F. 1962. A revised medium for rapid growth and boiassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.CrossRefGoogle Scholar
  23. Nielsen J.M., Kirsten B. & Hansen J. 1993. Long-term effects of thidiazuron is intermediate between benzyladenine, kinetin or isopentenyladenine in Miscanthus sinensis. Plant Cell Tiss Organ Cult. 35: 173–179.CrossRefGoogle Scholar
  24. Paterson A.H., Brubaker C.L. & Wendel J.F. 1993. A rapid method for extraction of cotton (Gossypium spp.) genomic DNA suitable for RFLP and PCR analysis. Plant Mol. Biol. Rep. 11: 112–127.CrossRefGoogle Scholar
  25. Price H.J. & Smith R.H. 1979. Somatic embryogenesis in suspension cultures of Gossypium klotzschianum Anderss. Planta. 145: 305–307.CrossRefGoogle Scholar
  26. Singh N.D., Sahoo, L., Bhalla N. & Jaiwal P.K. 2003. Effect of TDZ on organogenesis and somatic embryogenesis in pigeonpeas (Cajanus cajan L. Millsp). Plant Sci. 164: 341.CrossRefGoogle Scholar
  27. Sun Y.Q., Zhang X.L., Nie Y.C., Guo X.P., Jin S.X. & Liang S.G. 2004. Production and characterization of somatic hybrids between upland cotton (Gossypium hirsutum) and wild cotton (G. klotzschianum Anderss) via electrofusion. Theor. Appl. Genet. 109: 472–479.CrossRefPubMedGoogle Scholar
  28. Sun Y.Q., Zhang X.L., Huang C., Guo X.P. & Nie Y.C. 2006. Somatic embryogenesis and plant regeneration from different wild diploid cotton (Gossypium) species. Plant Cell Rep. 25: 289–296.CrossRefPubMedGoogle Scholar
  29. Tegeder M., Gebhardt D., Schieder O. & Pickardt T. 1995. Thidiazuron-induced plant regeneration from protoplasts of Vicia faba cv. Mythos. Plant Cell Rep. 15: 164–169.CrossRefGoogle Scholar
  30. Tzitzikas E.N., Bergervoet M., Raemakers K., Vincken J.P., Lammeren A.V. & Visser R.G.F. 2004. Regeneration of pea (Pisum sativum L.) by a cyclic organogenic system. Plant Cell Rep. 23: 453–460.CrossRefPubMedGoogle Scholar
  31. Yang X.Y., Gou X.P., Zhang X.L., Nie Y.C. & Jin S.X. 2007a. Plant regeneration from Gossypium davidsonii protoplastvia somatic embryogenesis. Biol. Plant. 51: 533–537.CrossRefGoogle Scholar
  32. Yang X.Y., Zhang X.L., Jin S.X., Fu L.L. & Wang L.G. 2007b. Production and characterization of asymmetric hybrids between upland cotton Coker 201 (Gossypium hirsutum) and wild cotton (G. klozschianum Anderss). Plant Cell Tiss Organ Cult. 89: 225–235.CrossRefGoogle Scholar

Copyright information

© © Versita Warsaw and Springer-Verlag Wien 2010

Authors and Affiliations

  • Xi-Yan Yang
    • 1
  • Xian-Long Zhang
    • 1
  • Li-Li Fu
    • 1
  • Ling Min
    • 1
  • Guan-ze Liu
    • 1
  1. 1.National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhanChina

Personalised recommendations