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Pea PR 10.1 is a Ribonuclease and its Transgenic Expression Elevates Cytokinin Levels
Authors:Sanjeeva Srivastava  R J Neil Emery  Leonid V Kurepin  David M Reid  Brian Fristensky  Nat N V Kav
Institution:(1) Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, T6G 2P5, Alberta, Canada;(2) Department of Biology, Trent University, Peterborough, K9J 7B8, Ontario, Canada;(3) Department of Biological Sciences, University of Calgary, Calgary, T2N 1N4 Alberta, Canada;(4) Department of Plant Science, University of Manitoba, Winnipeg, R3T 2N2, Manitoba, Canada
Abstract:The constitutive expression of a cDNA encoding a pea (Pisum sativum L.) PR 10 protein in Brassica napus leading to an enhancement of germination under saline conditions has been previously reported. In order to understand the biochemical function of this pea PR 10 protein, its cDNA has been expressed in Escherichia coli and the recombinant protein purified to homogeneity. Ribonuclease activity of the recombinant pea PR 10 protein has been demonstrated for the first time using an in-solution as well as an in-gel RNA degradation assay. Furthermore, in order to characterize the changes brought about as a result of the constitutive expression of the pea PR 10 cDNA in B. napus, we have measured the endogenous concentrations of several phytohormones. Increased cytokinin and, decreased abscisic acid (ABA) were observed in 7-day-old transgenic seedlings whereas no significant changes in the concentrations of gibberellin (GA) or indoleacetic acid (IAA) were observed at this stage of growth and development. The potential role(s) of PR 10 proteins with RNase activity and elevated cytokinins during plant stress responses as well as the possible relationship between PR 10 protein and changes in cytokinin concentrations are discussed.
Keywords:Brassica napus  Cytokinin  Pathogenesis related (PR) 10  Ribonuclease  tRNA
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