Mikroprojektil bombardımanı ile korunga (Onobrychis viciifolia Scop.) bitkisinde geçici beta-glukuronidaz reportör gen ifadesi

Transient $beta$-glucuronidase reporter gene (GUS) expression was visualised in whole cotyledons of sainfoin (Onobrychis viciifolia Scop.) after microprojectile bombardment with pBI221.23 DNA. Optimum rupture disk pressure and sample plate distance were determined to be 1350 psi and 6 cm, respectively. Mean gold particle diameter of 1.0 µm and 48 h post-bombardment histochemical GUS assay was found to be superior to 1.6 µm gold particles and 120 h post-bombardment GUS assay. Microscopic analysis revealed the localisation of the gold particles predominantly in the epidermal cell layer, whereas less than 1% of the particles were localised in sub-epidermal cells, as deep as three cell layers. Availability of a regeneration protocol from sainfoin cotyledons and the potential of the microprojectile bombardment recommend future applications for obtaining transgenic sainfoin plants.

Transient expression of beta-glucuronidase reporter gene in sainfoin (Onobrychis viciifolia Scop.) cotyledons via microprojectile bombardment

Transient $beta$-glucuronidase reporter gene (GUS) expression was visualised in whole cotyledons of sainfoin (Onobrychis viciifolia Scop.) after microprojectile bombardment with pBI221.23 DNA. Optimum rupture disk pressure and sample plate distance were determined to be 1350 psi and 6 cm, respectively. Mean gold particle diameter of 1.0 µm and 48 h post-bombardment histochemical GUS assay was found to be superior to 1.6 µm gold particles and 120 h post-bombardment GUS assay. Microscopic analysis revealed the localisation of the gold particles predominantly in the epidermal cell layer, whereas less than 1% of the particles were localised in sub-epidermal cells, as deep as three cell layers. Availability of a regeneration protocol from sainfoin cotyledons and the potential of the microprojectile bombardment recommend future applications for obtaining transgenic sainfoin plants.

___

  • 1. Christou, P., Swain, W.F., Yang, N.S., McCabe, D.E. Inheritance and expression of foreign genes in transgenic soybean plants, Proc. Natl. Acad. Sci. USA. 86, 7500-7504, 1989.
  • 2. Sato, S., Newell, C., Kolacz, K., Tredo, L., Finer, J., Hinchee, M. Stable transformation via particle bombardment in two different soybean regeneration systems, Plant Cell Rep. 12, 408-413, 1993.
  • 3. Warkentin, T.D., Jordan, M.C., Hobbs, S.L.A. Effect of promoter-leader sequences on transient reporter gene expression in particle bombarded pea (Pisum sativum L.) tissues, Plant Sci. 87, 171-177, 1992.
  • 4. Kar, S., Basu, D., Das, S., Ramkrishnan, N.A., Mukherjee, P., Nayak, P., Sen, S.K. Expression of CryIa(C) gene of Bacillus thuringiensis in transgenic chickpea plants inhibits development of pod borer (Heliothis armigera) larvae, Transgenic Res. 6, 177-185, 1997.
  • 5. Russell, D.R., Wallace, K.M., Bathe, J.H., Martinell, B.J., McCabe, D.E. Stable transformation of Phaseolus vulgaris via electric-discharge mediated particle acceleration, Plant Cell Rep. 12 , 165-169, 1992.
  • 6. Aragao, F.J.L., Grossi de Sa, M.F., Davey, M.R., Brasileiro, A.C.M., Faria, J.C., Rech, E.L. Factors influencing transient gene expression in bean (Phaseolus vulgaris L.) using an electric particle acceleration device, Plant Cell Rep. 12 , 483-490, 1993.
  • 7. Webb, K.J. Transformation of forage legumes using Agrobacterium tumefaciens, Theor. Appl. Genet. 72, 53-58, 1986.
  • 8. Klein, T.M., Wolf, E.D., Wu, R., Sanford, J.C. High velocity microprojectiles for delivering nucleic acids into living cells, Nature 327, 70-73, 1987.
  • 9. Christou, P. Genetic transformation of crop plants using microprojectile bombardment, Plant J. 2, 275-281, 1992.
  • 10. Klien, T.M., Arentzen, R., Lewis, P.A., Fitzpatrick-McElligott, S. Transformation of microbes, plants and animals by particle bombardment, Bio/Technology 10, 286-291, 1992.
  • 11. Luthra, R., Varsha Dubey, R.K., Srivastava, A., Kumar, S. Microprojectile mediated plant transformation-A bibliographic search, Euphytica 95, 269-294, 1997.
  • 12. Sanford, J.C., Smith, F.D., Russell, J.A. Optimizing the biolistic process for different biological applications, Methods Enzymol. 217, 483-509, 1993.
  • 13. Murashige, T., Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures, Physiol. Plant. 15, 473-497, 1962.
  • 14. Lonsdale, D., Önde, S., Cuming, A.C. Transient expression of exogenous DNA in intact, viable wheat embryos following particle bombardment, J. Exp. Bot. 4, 1161-1165, 1990.
  • 15. Meyerowitz, E.M. In situ hybridisation to RNA in plant tissues, Plant Mol. Biol. Rep. 5, 242-250, 1987.
  • 16. Özcan, S., Yıldız, M., Sancak, C., Özgen, M. Adventitious shoot regeneration in sainfoin (Onobrychis viciifolia Scop.), Tr. J. Botany 20, 497-501, 1996.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK