Secretion and efflux of oxalic acid from roots is an important aluminum detoxification mechanism for various plants; however, how this process is completed remains unclear. In this study, the candidate oxalate transporter gene AtOT, encoding 287 amino acids, was cloned and identified from Arabidopsis thaliana. AtOT was upregulated in response to aluminum stress at the transcriptional level, which was closely related to aluminum treatment concentration and time. The root growth of Arabidopsis was inhibited after knocking out AtOT, and this effect was amplified by aluminum stress. Yeast cells expressing AtOT enhanced oxalic acid resistance and aluminum tolerance, which was closely correlated with the secretion of oxalic acid by membrane vesicle transport. Collectively, these results underline an external exclusion mechanism of oxalate involving AtOT to enhance oxalic acid resistance and aluminum tolerance.

Aluminum (Al) toxicity severely restricts crop growth and productivity in acidic soils. The rubber tree is one of the most economically important crops in tropical regions, which is tolerant to high concentrations of Al in sand or hydroponic culture conditions compared with other plants that have been reported. However, the mechanisms of Al tolerance in rubber trees remain unknown. In this study, we conducted a transcriptome and metabolome analysis for rubber tree sapling roots treated with 200 mM Al for 0 (CK), 2 or 5 days, respectively. Compared with the CK, a total of 9534 differentially expressed genes (DEGs) and 3821 differentially expressed metabolites (DEMs) were identified in 2 d of Al treatment. There were 10,373 DEGs and 4636 DEMs after 5 d of Al treatment, and 1626 DEGs and 1674 DEMs between 2 and 5 d of Al treatment. The DEGs mainly concentrated in transporters, transcription factors (TFs), cell wall biosynthesis and antioxidant systems, and the DEMs were mainly focused on lipids and lipid-like molecules, organic acids and derivatives, organic oxygen compounds, phenylpropanoids and polyketides. The combined transcriptome and metabolome analysis indicated DEGs and DEMs involved in ABC transporters, glutathione metabolism, flavonoid biosynthesis and phenylalanine metabolic pathways were identified to be closely associated with the Al tolerance of rubber trees. Our study elucidated the mechanism of rubber trees’ tolerance to Al at the transcriptional and metabolic levels, which provides a theoretical basis for the study of Al tolerance both for rubber trees and other woody plants.

Secretion of oxalic acid from roots is an important aluminum detoxification mechanism for many plants such as Hevea brasiliensis (rubber tree). However, the underlying molecular mechanism and oxalate transporter genes in plants have not yet been reported. In this study, the oxalate transporter candidate genes HbOT1 and HbOT2 from the rubber tree were cloned and preliminarily identified. It was found that HbOT1 had a full length of 1163 bp with CDS size of 792 bp, encoding 263 amino acids, and HbOT2 had a full length of 1647 bp with a CDS region length of 840 bp, encoding 279 amino acid residues. HbOT1 and HbOT2 were both stable hydrophobic proteins with transmembrane structure and SNARE_assoc domains, possibly belonging to the SNARE_assoc subfamily proteins of the SNARE superfamily. qRT-PCR assays revealed that HbOT1 and HbOT2 were constitutively expressed in different tissues, with HbOT1 highly expressed in roots, stems, barks, and latex, while HbOT2 was highly expressed in latex. In addition, the expressions of HbOT1 and HbOT2 were up-regulated in response to aluminum stress, and they were inducible by metals, such as copper and manganese. Heterologous expression of HbOT1 and HbOT2 in the yeast mutant AD12345678 enhanced the tolerance to oxalic acid and high concentration aluminum stress, which was closely correlated with the secretion of oxalic acid. This study is the first report on oxalate transporter genes in plants, which provides a theoretical reference for the study on the molecular mechanism of oxalic acid secretion to relieve aluminum toxicity and on aluminum-tolerance genetic engineering breeding.