Jm. Humara et al., Improved efficiency of uidA gene transfer in stone pine (Pinus pinea) cotyledons using a modified binary vector, CAN J FORES, 29(10), 1999, pp. 1627-1632
Citations number
28
Categorie Soggetti
Plant Sciences
Journal title
CANADIAN JOURNAL OF FOREST RESEARCH-REVUE CANADIENNE DE RECHERCHE FORESTIERE
This paper describes the construction of a binary vector with the uidA-int
gene under the control of the promoter and first intron from the maize (Zea
mays L.) ubiquitin 1 (Ubi1) gene, aiming at high expression levels of the
uidA gene in Pinus pinea L. cotyledons. Transient expression was determined
after microparticle bombardment of 1-day-old cotyledons, and data indicate
d that the new construct performed considerably better than p35SGUSint, whi
ch has the uidA-int gene under the control of a CaMV35S promoter. In additi
on, we have demonstrated that the marker gene uidA from plasmid pBINUbiGUSi
nt is reproducibly transferred from Agrobacterium tumefaciens (E.F. Sm. & T
own) Conn, and beta-glucuronidase activity was detected 7 days after inocul
ation of scraped or sonicated cotyledons. Infection with A. tumefaciens EHA
105 pBINUbiGUSint significantly improved results obtained earlier with plas
mid p35SGUSint. Further, the application of our protocol reduced the level
of the necrotic response that is usually displayed by cotyledons after Agro
bacterium infection.