In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

  • Hafiz Mamoon REHMAN Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, 38040, Faisalabad
  • Iqrar Ahmad RANA Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, 38040, Faisalabad
  • Siddra IJAZ Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, 38040, Faisalabad
  • Ghulam MUSTAFA Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, 38040, Faisalabad
  • Faiz Ahmad JOYIA Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, 38040, Faisalabad
  • Iqrar Ahmad KHAN Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, 38040, Faisalabad
  • Paula M. PIJUT USDA Forest Service, Northern Research Station, Hardwood Tree Improvement and Regeneration Center, 715 West State Street, West Lafayette, Indiana 47907

Abstract

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.

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Published
2012-11-05
How to Cite
REHMAN, H. M., RANA, I. A., IJAZ, S., MUSTAFA, G., JOYIA, F. A., KHAN, I. A., & PIJUT, P. M. (2012). In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 40(2), 140-147. https://doi.org/10.15835/nbha4028248
Section
Research Articles