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Expression of a cotton fiber "specific" gene promoter in tobacco and cotton

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Date
1996-05
Author
Dang, Phat Minh
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Abstract
Cotton fibers {Gossypiuiv hirsutum L.) are highly elongated trichomes that grow from epidermal cells of cotton ovules. Analysis of genes that are specifically expressed during fiber differentiation can provide insights into the molecular events that control cotton fiber development. One such gene, called Gh-1, has been isolated and characterized. Deletion analysis of the 5' flanking sequences of the Gh-1 gene was performed to identify the putative regulatory elements responsible for fiber specific expression. The Gh-1 promoter, along with various lengths of 5' flanking sequences, were fused with the p-glucuronidase (GUS) reporter gene and transferred into tobacco and cotton plants via /\gro/)acter/aAr7-mediated transformation. Tobacco plants that carry the Gh-1::GUS gene construct with 965 bp of 5' flanking sequence (-965) from the putative transcription start site typically showed very weak GUS activity in vascular tissues and guard cells, and, in some plants, expression was seen in small glandular trichomes. Similar expression patterns were also seen in plants that carried -735, and -635 Gh-1::GUS promoter constructs. A dramatic increase in specific expression in large glandular trichomes was observed in plants that carried a -500 Gh-1::GUS promoter construct. Promoter deletion constructs -305 Gh-1::GUS or shorter showed no observable GUS staining. Cotton plants single cell line #41 carrying the -965 Gh-1::GUS construct showed no observable GUS staining in stem and 0-3 DPA ovules. However, promoter deletion -500 Gh-1::GUS construct single cell line #22 showed strong GUS staining in guard cells and glandular trichomes in the stem and staining in 0-3 DPA ovules. These results indicated that a trichome and guard cell "specific" regulatory element apparently exist between -500 and -305 of the Gh-1 promoter.
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http://hdl.handle.net/2346/14351
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