Agrobacterium rhizogenes-mediated DNA transfer in Pinus halepensis Mill

Tzvi Tzfira*, Ophir Yarnitzky, Alexander Vainstein, Arie Altman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

54 Scopus citations

Abstract

Agrobacterium rhizogenes strain LBA9402 was used to transform Pinus halepensis embryos, seedlings and shoots. Mature embryos exhibited susceptibility to the agrobacterium as monitored by β-glucuronidase (GUS) expression, with more than 85% showing considerable transient GUS expression in the radicle. GUS expression was also observed in cotyledons, but at a lower rate of about 24% of the embryos (1-5 spots/embryo). Stable transformation was evidenced by the regeneration of GUS-expressing roots and calli from infected P. halepensis seedlings. Inoculum injections into intact seedling hypocotyls induced callus and root formation at the wound sites in 64% of the seedlings. Dipping seedling cuttings in a bacterial suspension resulted in adventitious root formation in 71% of the seedling cuttings, all of which expressed GUS activity. Adventitious shoots, that were induced on 2.5-year-old seedlings by pruning and spraying with 6-benzylaminopurine, were infected by injecting of bacterial suspension into their basal side. Two months later, adventitious roots and root primordia regenerated in 74% and 40% of 2- and 5-month-old shoots, respectively. Non-transformed shoots, either without or with auxin application, failed to form roots. Polymerase chain reaction and Southern blot analyses confirmed the uidA-transgenic nature of the root and callus, as well as the presence of rolC and rolB genes in roots from infected P. halepensis seedlings.

Original languageAmerican English
Pages (from-to)26-31
Number of pages6
JournalPlant Cell Reports
Volume16
Issue number1-2
DOIs
StatePublished - 1996

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