Molecular cloning, characterization and regulation of two different NADH-glutamate synthase cDNAs in bean nodules

Plant, Cell & Environment
Lourdes BlancoMiguel Lara-Flores

Abstract

NADH-dependent glutamate synthase (NADH-GOGAT) is a key enzyme in primary ammonia assimilation in Phaseolus vulgaris nodules. Two different types of cDNA clones of PvNADH-GOGAT were isolated from the nodule cDNA libraries. The full-length cDNA clones of PvNADH-GOGAT-I (7.4 kb) and PvNADH-GOGAT-II (7.0 kb), which displayed an 83% homology between them, were isolated using cDNA library screening, 'cDNA library walking' and RT-PCR amplification. Southern analysis employing specific 5' cDNA probes derived from PvNADH-GOGAT-I and PvNADH-GOGAT-II indicated the existence of a single copy of each gene in the bean genome. Both these proteins contain approximately 100 amino acid sequences theoretically addressing each isoenzyme to different subcellular compartments. RT-PCR analysis indicated that PvNADH-GOGAT-II expression is higher than PvNADH-GOGAT-I during nodule development. Expression analysis by RT-PCR also revealed that both of these genes are differentially regulated by sucrose. On the other hand, the expression of PvNADH-GOGAT-I, but not PvNADH-GOGAT-II, was inhibited with nitrogen compounds. In situ hybridization and promoter expression analyses demonstrated that the NADH-GOGAT-I and -II genes are differentially expressed in be...Continue Reading

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Citations

Dec 24, 2013·Plant Physiology and Biochemistry : PPB·Jacob Pérez-TiendaNuria Ferrol
Dec 3, 2015·Plant & Cell Physiology·Laure GaufichonAkira Suzuki
Nov 6, 2012·The New Phytologist·Aarón BarrazaFederico Sanchez

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