ISSN:
1432-203X
Keywords:
Abbreviations: BA, 6-benzylaminopurine; NOS, nopaline synthase; PCR, polymerase chain reaction; EtOH, ethanol; GUS, β-glucuronidase; NPTII, neomycin phosphotransferase II; CaMV, califlower mosaic virus; X-gluc, 5-bromo-4-chloro-3-indolyl β-D-glucuronic acid.
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
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 49% 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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s002990050169
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