Stable transformation of mature zygotic embryos and regeneration of transgenic plants of chir pine (Pinus roxbughii Sarg.).

Autor: Parasharami VA; Tissue Culture Pilot Plant, National Chemical Laboratory, Dr. Homi Bhabha Road, Pashan, Pune, 411 008, Maharashtra, India. varshapp2@yahoo.com, Naik VB, von Arnold S, Nadgauda RS, Clapham DH
Jazyk: angličtina
Zdroj: Plant cell reports [Plant Cell Rep] 2006 Jan; Vol. 24 (12), pp. 708-14. Date of Electronic Publication: 2005 Aug 17.
DOI: 10.1007/s00299-005-0019-z
Abstrakt: A particle inflow gun was used to transfer the plasmid pAHC25 containing the bar gene conferring resistance to glufosinate and the gusA reporter gene, each driven by the maize ubiquitin promoter, to mature embryos of Pinus roxburghii (chir pine). High levels of transient expression were obtained when embryos were cultured for 6 days on 10 microM benzyl adenine-containing medium and then exposed to high osmoticum (0.5 M sucrose) before and after bombardment. Selection on medium containing Basta enabled recovery of stably transformed shoots, both from the epicotyl and from adventitious buds. The primary transformed shoots from the epicotyl were multiplied via axillary shoots. Transformation was confirmed by histochemical staining for beta-glucuronidase (GUS) activity, by polymerase chain reaction (PCR) amplification of fragments of gusA and nos terminator, and by the resistance of needles to Basta.
Databáze: MEDLINE