Expression of Bacterial L-aspartate-alpha-decarboxylase in Tobacco Increases Beta-alanine and Pantothenate Levels and Improves Thermotolerance

Overview

Journal Plant Mol Biol

Date 2006 Mar 10

PMID 16525887

Citations 15

Authors

Walid M Fouad

Bala Rathinasabapathi

Affiliations

Soon will be listed here.

Abstract

L- Aspartate-alpha-decarboxylase catalyzes the decarboxylation of L -aspartate to generate Beta-alanine and carbon dioxide. This is an unusual pyruvoyl-dependent enzyme unique to prokaryotes that undergoes limited self-processing. The Escherichia coli pan D gene encoding L- aspartate-alpha-decarboxylase was expressed under a constitutive promoter in transgenic tobacco. Transgene expression was verified by assays based on RNA blots, immunoblots and enzyme activity in vitro. The pan D lines had increased levels of leaf Beta-alanine (1.2- to 4-fold), pantothenate (3.2- to 4.1-fold) and total free amino acids (up to 3.7-fold) compared to wild-type and vector controls. Growth of homozygous lines expressing E. coli L- aspartate-alpha-decarboxylase was less affected than that of the control lines when the plants were stressed for 1 week at 35 degrees C. When transferred from 35 to 30 degrees C for 3 weeks, the Pan D transgenic lines recovered significantly (P <or= 0.001) better than the controls: Pan D lines had on an average 54% and 84% greater fresh and dry weights respectively, compared to the controls. Homozygous lines expressing E. coli L- aspartate-alpha-decarboxylase had significantly greater thermotolerance (P<or=0.05) during germination. At 42 degrees C, 95% of two T3 Pan D transgenic line seeds germinated after 12 days compared to 73% for the wild-type seeds. Our results indicated that E. coli L-aspartate-alpha-decarboxylase was correctly processed and active in the transgenic eukaryotic host and its expression resulted in increased thermotolerance in tobacco.

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