Evaluation of promoters and visual markers for transformation of eastern white pine
Source: New Forests, Volume 21, Number 1, January 2001 , pp. 45-58(14)
Abstract:This report serves to evaluate possible promoters for use in the production of transgenic eastern white pine (Pinus strobus L.). Embryogenic cultures of eastern white pine were bombarded with gold particles coated separately with a variety of gene constructs containing the UidA -glucuronidase (GUS) or green fluorescent protein (GFP) reporter gene. Transient expression of the UidA gene, driven by a novel algal virus adenine methyl transferase gene promoter, as well as five other promoters used in angiosperm transformation, were evaluated. The maize alcohol dehydrogenase promoter was not effective in eastern white pine cultures. The construct with the doubled Cauliflower Mosaic Virus 35S promoter plus Alfalfa Mosaic Virus enhancer showed the highest levels of expression. GUS expression was detected within 24 hours, but decreased after 5 days and was not detectable 15 days after bombardment. Expression of GUS activity was recorded mainly in somatic embryonal heads of various stages of development and occasionally in suspensor cells. Similar to GUS expression, modified green fluorescent protein (GFP) was detected in the embryonal head cells 24 hours after bombardment. GFP-expressing suspensor cells were both more infrequent and difficult to detect, as their highly vacuolate nature rendered the GFP presence less visible against the yellow background autofluorescence.
Document Type: Regular Paper
Affiliations: 1: Department of Plant and Soil Science, Alabama A&M University, Normal, AL 35762, USA (e-mail: email@example.com 2: USDA Forest Service, Southern Institute of Forest Genetics, Gainesville, FL 32611, USA 3: Department of Plant and Soil Science, Alabama A&M University, Normal, AL 35762, USA 4: Department of Surgery, University of Alabama-Birmingham, Birmingham, AL 35294, USA 5: Department of Botany, University of Rajasthan, India
Publication date: January 2001