Gene silencing studies in the gymnosperm species Pinus radiata

Armin Wagner*, Lorelle Phillips, Reena D. Narayan, Judy M. Moody, Barbara Geddes

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

A biolistic transformation procedure was used to transform embryogenic Pinus radiata tissue with constructs containing the Zea mays UBI1 (ubiquitin)-promoter followed by the P. radiata CAD (cinnamyl alcohol dehydrogenase) cDNA in sense or anti-sense orientation or in the form of an inverted-repeat. The effect of the different constructs on silencing the endogenous CAD gene was monitored in embryogenic tissue and somatic seedlings of 28 P. radiata transclones. Quantitative CAD measurements demonstrated that the construct containing an inverted-repeat of the CAD cDNA was most efficient in triggering gene silencing in P. radiata. Northern hybridization experiments with silenced transclones revealed that reduced CAD activities were the result of reduced steady state levels of the targeted CAD mRNA. Monitoring of the activity of the UBI1-promoter in the P. radiata transclones and heat-shock experiments with transgenic somatic P. radiata seedlings indicated that gene silencing is positively correlated with the expression level of the transgene. The obtained data are also consistent with a role for the expression level of the endogenous CAD gene in gene silencing.

Original languageEnglish
Pages (from-to)95-102
Number of pages8
JournalPlant Cell Reports
Volume24
Issue number2
DOIs
Publication statusPublished - May 2005

Fingerprint

Dive into the research topics of 'Gene silencing studies in the gymnosperm species Pinus radiata'. Together they form a unique fingerprint.

Cite this