Regeneration and Transformation Experiments in Apple
Successful introduction of foreign genes using a leaf disc procedure and cocultivation with Aqrobacterium tumefaciens requires a good regeneration protocol and tissue susceptible to infection. In our experiments, apple rootstocks M26 and M9 and the cultivar Mcintosh Wijcik were used. A good regeneration system was developed for M9 whereas a suitable protocol for the others had been previously developed. Transformation experiments were carried out with three wild type strains and three disarmed vectors harbouring ß-glucuronidase and neomycin phosphotransferase genes. Infection with three wild type strains (C58, A348, A281) did not cause gall formation. However, the binary vector pBH21 (35SGUS) x LBA 4404 resulted in transformed shoots of M26 and transformed callus of M9. This was confirmed histochemically using the substrate 5-bromo-4-chloro-3-indolyl glucoronide (X-Gluc) staining. Cefotaxime, used to control growth of the bacteria, had an effect simalar to cyto-kinin by stimulating shoot formation in vitro.
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- 1.Alston, F.H., 1970, Resistance to collar rot. Phytophthora cactorum (Leb.and Cohn) Schroet.,in apple. Rep. E.Mailing Res. Stn. for 1969: 143–145.Google Scholar
- 3.Chu, C.C., Wang, C.C., Sun, C.S., Hsü, C., Yin, K.C., Chu, C.Y., and Bi, F.Y., 1975, Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci. Sin. 18: 659–668.Google Scholar
- 11.Knight, R.L., and Alston, F.H., 1969, Developments in apple breeding. Rep. E.Mailing Res. Stn. for 1968: 125–132.Google Scholar
- 16.Pasqualetto, P.L., Zimmerman R.H., and Fordham, I., 1986, Gelling agent and growth regulator effects on shoot vitrification of “Gala” apple in vitro. J. Amer. Soc. Hort. Sci. 111(6): 976–980.Google Scholar
- 17.Pierik, R.L.M., 1971, Plant tissue culture as motivation for the symposium. Misc. Pap. Landbouwhogesch. Wageningen. 9: 3–13.Google Scholar
- 19.Shay, J.R., Dayton, D.F., and Hough, L.F., 1953, Apple scab resistance from a number of Malus species. Proc. Soc. Hort. Sci. 62: 348–356.Google Scholar