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Title: CHANGES IN DEHYDRIN EXPRESSION ASSOCIATED WITH COLD, ABA AND PEG TREATMENTSIN BLUEBERRY CELL CULTURES

Author
item PARMENTIER-LINE, CECILE - UNIV OF MARYLAND
item PANTA, GANESH - UNIV OF TENNESSEE
item Rowland, Lisa

Submitted to: Plant Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/25/2001
Publication Date: 2/20/2002
Citation: Parmentier-Line, C.M., Panta, G.R., Rowland, L.J. 2002. Changes in dehydrin expression associated with cold, aba and peg treatmentsin blueberry cell cultures. Plant Science.

Interpretive Summary: Because the development of more cold hardy varieties is an important need to the blueberry industry, our laboratory has been working on identifying genes that control cold hardiness in blueberry. We have identified three abundant proteins that are induced during cold acclimation and their accumulation is closely associated with cold hardiness levels. These proteins are a type of plant protein induced by cold and drought stress, known as dehydrin, and are good candidates for controlling cold hardiness in blueberry. Carrying out gene expression studies to understand how these genes are expressed and regulated can be difficult in whole plants especially woody plants. Cell culture systems offer some advantages over whole plants for gene expression studies such as allowing more control over certain types of treatments and allowing experiments to be carried out year-round. Therefore, we have investigated whether cell cultures can be used instead of whole plants for studying dehydrin expression in blueberry. In this study, we examined their expression in blueberry cell cultures in response to several treatments including cold and polyethylene glycol, which induces water stress, and expression was compared to that seen previously with whole plants. The results indicated that cultures responded differently from whole plants. Therefore cell culture is not a good system for studying dehydrin expression in blueberry. The study of genes associated with cold hardiness will help scientists in developing new, more cold hardy varieties of blueberry.

Technical Abstract: Cell cultures of highbush blueberry cultivar Gulfcoast (Vaccinium corymbosum L. x V. darrowi Camp) were used to determine if tissue cultures could be used as a surrogate for whole plants in studying dehydrin expression. Cell clusters in liquid medium were subjected to treatments known to induce dehydrins in whole plants, and dehydrin protein and RNA levels were monitored. Two dehydrins of 65 and 30 kDa were detected with a polyclonal antibody raised against the 65 kDa dehydrin of blueberry. Using a full length cDNA clone of blueberry dehydrin 1 as a probe, one mRNA of 0.75 kb, an appropriate size to encode the 30 kDa dehydrin, was detected on RNA blots. When cells were grown at 4C, abundance of the 30 kDa dehydrin increased, but the 65 kDa dehydrin did not. Level of the 0.75 kb message increased temporarily then decreased with cold treatment. Disparities between protein and transcript levels suggested that post-transcriptional mechanisms were involved in regulation of dehydrin expression. Some of the ABA concentrations tested induced expression of both dehydrins. Polyethylene glycol in the cultures repressed expression of both dehydrins and of the 0.75 kb mRNA. Results obtained from cell cultures differed in too many ways from those on whole plants to make cell culture suitable for studying dehydrin expression in blueberry.