An abscisic acid (ABA) homeostasis regulated by its production, catabolism and transport in peanut leaves in response to drought stress.

ABA is an important messenger that acts as a signaling mediator for regulating the adaptive response of plants to drought stress. Two production pathways, de novo biosynthesis and hydrolysis of glucose-conjugated ABA by β-glucosidase (BG), increase cellular ABA levels in plants. ABA catabolism via hydroxylation by 8'-hydroxylase (CYP707A), or conjugation by uridine diphosphate glucosyltransferase (UGT), decreases cellular ABA levels. The transport of ABA through ATP-binding cassette (ABC)-containing transporter proteins, members of ABC transporter G family (ABCG), across plasma membrane (PM) is another important pathway to regulate cellular ABA levels. In this study, based on our previously constructed transcriptome of peanut leaves in response to drought stress, fourteen candidate genes involved in ABA production (including AhZEP, AhNCED1 and AhNCED3, AhABA2, AhAAO1 and AhAAO2, AhABA3, AhBG11 and AhBG24), catabolism (including AhCYP707A3, AhUGT71K1 and AhUGT73B4) and transport (including AhABCG22-1 and AhABCG22-2), were identified homologously and phylogenetically, and further analyzed at the transcriptional level by real-time RT-PCR, simultaneously determining ABA levels in peanut leaves in response to drought. The high sequence identity and very similar subcellular localization of the proteins deduced from 14 identified genes involved in ABA production, catabolism and transport with the reported corresponding enzymes in databases suggest their similar roles in regulating cellular ABA levels. The expression analysis showed that the transcripts of AhZEP, AhNCED1, AhAAO2 and AhABA3 instead of AhABA2, AhNCED3 and AhAAO1 in peanut leaves increased significantly in response to drought stress; and that the AhBG11 and AhBG24 mRNA levels were rapidly and significantly up-regulated, with a 4.83- and 4.58-fold increase, respectively at 2-h of drought stress. The genes involved in ABA catabolism AhCYP707A3, AhUGT71K1 instead of AhUGT73B4 were significantly induced in response to drought stress. The expression of two closely related peanut ABCG genes, AhABCG22.1 and AhABCG22.2, was significantly up-regulated in response to drought stress. The ABA levels rapidly began to accumulate within 2 h (a 56.6-fold increase) from the start of drought stress, and peaked at 10 h of the stress. The highly and rapidly stress up-regulated expressions of genes involved in ABA production and transport, particularly AhNCED1, AhBG11 and AhBG24, and AhABCG22.1 and AhABCG22.2, might contribute to the rapid ABA accumulation in peanut leaves in response to drought. In response to drought stress, ABA accumulation levels in peanut leaves agree well with the up-regulated expressions of ABA-producing genes (AhZEP, AhNCED1, AhAAO2, AhABA3, AhBG11 and AhBG24) and PM-localized ABA importer genes (AhABCG22-1 and AhABCG22-2), in spite of the simultaneously induced ABA catabolic genes (AhCYP707A3 and AhUGT71K1), although the induction of catabolic genes was much lower than that of biosynthetic gene (AhNCED1). This difference in induction kinetics of gene expression may define the significant accumulation of drought-induced ABA levels. These results suggest that ABA homeostasis in peanut leaves in response to drought maintained through a balance between the production, catabolism and transport, rather than simply by the biosynthesis.