Comparative proteomic analysis of gamma-aminobutyric acid responses in hypoxia-treated and untreated melon roots.

Hypoxia is one of the main environmental stresses that accounts for decreasing crop yield. To further investigate the mechanisms whereby exogenous GABA alleviates hypoxia injury to melon seedlings, a comparative proteomic analysis was performed using roots subjected to normal aeration and hypoxia conditions with or without GABA (5mM). The results indicated that protein spots on gels after hypoxia and hypoxia+GABA treatment were significantly changed. Three "matched sets" were analyzed from four treatments, and 13 protein spots with large significant differences in expression were identified by MALDI-TOF/TOF mass spectrometry. Exogenous GABA treatment enhanced the expression of protein in cytosolic phosphoglycerate kinase 1, exaA2 gene product, dnaJ and myb-like DNA-binding domain-containing proteins, as well as elongation factor-1 alpha and hypothetical proteins in hypoxia-induced roots. However, the hypoxia+GABA treated roots had a significantly lower expression of proteins including malate dehydrogenase, nucleoside diphosphate kinase, disease resistance-like protein, disulfide isomerase, actin, ferrodoxin NADP oxidoreductase, glutathione transferase, netting associated peroxidase. This paper describes the effect of GABA on melon plants under hypoxia-induced stress using proteomics, and supports the alleviating function of GABA in melon plants grown under hypoxic conditions.

Polyamine biosynthesis and degradation are modulated by exogenous gamma-aminobutyric acid in root-zone hypoxia-stressed melon roots.

We detected physiological change and gene expression related to PA metabolism in melon roots under controlled and hypoxic conditions with or without 5 mM GABA. Roots with hypoxia treatment showed a significant increase in glutamate decarboxylase (GAD) activity and endogenous GABA concentration. Concurrently, PA biosynthesis and degradation accelerated with higher gene expression and enzymes activity. However, endogenous GABA concentrations showed a large and rapid increase in Hypoxia + GABA treated roots. This led to a marked increase in Glu concentration by feedback inhibition of GAD activity. Hypoxia + GABA treatment enhanced arginine (Arg), ornithine (Orn) and methionine (Met) levels, promoting enzyme gene expression levels and arginine decarboxylase (ADC), ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) activities in roots. Hypoxia + GABA treatment significantly increased concentrations of free putrescine (Put), spermidine (Spd) and spermine (Spm) from day two to eight, promoting the PA conversion to soluble conjugated and insoluble bound forms. However, PA degradation was significantly inhibited in hypoxia + GABA treated roots by significantly decreasing gene expression and activity of diamine oxidase (DAO) and polyamine oxidase (PAO). However, exogenous GABA showed a reduced effect in control compared with hypoxic conditions. Our data suggest that alleviating effect of exogenous GABA to hypoxia is closely associated with physiological regulation of PA metabolism. We propose a potential negative feedback mechanism of higher endogenous GABA levels from combined effects of hypoxia and exogenous GABA, which alleviate the hypoxia damage by accelerating PA biosynthesis and conversion as well as preventing PA degradation in melon plants.

[Effects of exogenous gamma-aminobutyric acid on polyamine metabolism of melon seedlings under hypoxia stress].

Taking melon cultivar 'Xiyu No. 1 ' as test material, a hydroponic experiment was conducted to investigate the effects of exogenous gamma-aminobutyric acid (GABA) on the seedlings polyamine metabolism under hypoxia stress. Compared with the control in normoxic treatment, the seedlings under hypoxia stress had significantly higher glutamic acid decarboxylase (GAD) activity and GABA content, and their polyamine synthesis enzymes activities all enhanced significantly, which led to a marked increase of polyamines contents. Meanwhile, the seedlings leaf- and root diamine oxidase (DAO) and polyamine oxidase (PAO) activities also had a significant increase. The increment of root arginine decarboxylase (ADC) activity was higher, which induced a higher content of free putrescine (Put) in roots, while the increment of leaf ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) activities were higher, inducing a higher content of free spermidine (Spd) in leaves. The PBs-extractable DAO and PAO activies in roots were significantly lower than those in leaves, but the cell wall-bound PAO activity was in adverse. Under hypoxia stress, the addition of exogenous GABA increased the leaf- and root GABA and glutamic acid contents and decreased the GAD activity significantly. The increase of arginine, ornithine, and methionine contents promoted the activities of polyamines synthesis enzymes, which led to the significant increase of polyamines contents and the significant decrease of DAO and PAO activities.