Expression of bacterial L-aspartate-alpha-decarboxylase in tobacco increases beta-alanine and pantothenate levels and improves thermotolerance.
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.
Fouad
Walid M
WM
Plant Molecular and Cellular Biology Program, Horticultural Sciences Department, University of Florida, Gainesville, FL 32611-0690, USA.
Rathinasabapathi
Bala
B
D000064:Acclimatization / Q000235:genetics / Q000502:physiology*; D018533:Biomass; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics; D018525:Germination / Q000502:physiology; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism*; D010205:Pantothenic Acid / Q000378:metabolism*; D010641:Phenotype; D018515:Plant Leaves / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000502:physiology; D013696:Temperature; D014026:Nicotiana / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology; D015091:beta-Alanine / Q000378:metabolism*
D012333:RNA, Messenger; D015091:beta-Alanine; D010205:Pantothenic Acid; C019104:aspartate-alpha-decarboxylase; D005968:Glutamate Decarboxylase