Preliminary studies on establishment of genetic transformation system in loblolly pine

Abstract

A transformation system was established for loblolly pine (Pinus taeda L.) mature zygotic embryos usingAgrobacterium tumefaciens. The gene coding for the β-glucuronidase (GUS) gene was introduced into loblolly pine tissues and its transient expression was detected with histochemical staining. The influences of different genotypes.Agrobacterum concentrations. and cocultivation time on GUS expression and kanamycin resistant callus and shoot regeneration were investigated. The results showed that the highest GUS expression frequency (16.3%) and shoot regeneration frequency (78%) were obtained from genotype 9–1003 withAgrobacterium concentration decreased 9 times and cocultivation time of 56 hours respectively. GUS expression was obtained in all genotypes tested. The successful expression of the GUS gene in different genotypes suggested that it will be a useful transformation system for loblolly pine.

Keywords

Pinus taeda L. / Genetic transformation / Agrobacterium tumefaciens

References

[1] Ellis D., Roberts D., Sutton B.. Transformation of white spruce and other conifer species byAgrobacterium tumefaciens. Plant Cell Rep., 1989, 8: 16-20.

[2] Loopstra C.A., Stomp A.M., Sederoff R.. Agrobacterium-mediated DNA transfer in sugar pine. Plant Mol. Biol, 1990, 15: 1-9.

[3] Sederoff R., Stomp A.M., Chilton W.S.. Gene transfer into loblolly pine byAgrobacterium tumefactens. Bin/Technology, 1986, 4: 647-649.

[4] Jouanin I., Brasileiro A.M., Leple J.C.. Genetic transformation: a short review of methods and their applications results and perspectives for fores trees. Ann Sci. For., 1993, 50: 325-336.

[5] Ellis D., McCabe D.E., Meinnis S.. Stable transformation ofPicea glanca by particle acceleration. Bio/Technology, 1993, 11: 84-89.

[6] Levee V., Lelu A.M., Jouanin L.. Agrobacterium tumefaciens-mediated transformation of hybrid larch and transgenic plant regeneration. Plant Cell Rep., 1997, 16: 680-685.

[7] Gupta P.K., Dandekar A.M., Durzan D.J.. Somatic proembryo formation and transient expression of a luciferase gene in Douglas fir and loblolly pine pretoplast. Plant Sci., 1988, 58: 85-92.

[8] Robertson, D., Weissinger, A.K., and Ackley. R. 1992. Genetic transformation of Norway spruce using somatic embry. explants by microprojectile bombardment. Plant Mol. Biol. 925–935

[9] Charest P.J., Devantier Y., Lachance D.. Stable genetic transformation ofPicea mariana via microprojectile bombardment. In Vitro Cell Dev. Brol., 1996, 32: 91-99.

[10] Gupta P.K., Durzan D.J.. Somatic polyembryogenesis from callus of mature supar pine embryos. Bio/Technology, 1986, 4: 643-645.

[11] Jefferson R.A.. Assaying chimeric genes in plants: the GUS fusion system. Plant Mol. Biol. Res., 1987, 5(4): 387-405.

[12] Cheng M., Jarret R.L., Li Z.. Production of fertile transgenic peanut plants usingAgrobacterium tumefaciens. Plant Cell Rep., 1996, 15: 653-657.

[13] Tautorus T.E., Fowke L.C., Dunstan D.I.. Somatic embryogenesis in conifers. Can. J. Bot., 1991, 69: 1876-1899.

[14] Ellis D., McCabe D., Russell D.. Expression of inducible angiosperm promoters in a gymnosperm.Picea glauca. Plant Mol. Biol., 1991, 17: 19-27.

[15] Tian L., Seguin A., Charest P.J.. Expression of the green fluorescent protein gene in conifer tissues. Plant Cell Rep., 1997, 16: 267-271.

[16] Duchesne L.C., Charest P.J.. Transient expression of the β-glucuronidase gene in embryogenic callus ofPicea mariana following microprojection. Plant Cell Rep., 1991, 10: 191-194.