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.

Plant 14-3-3 proteins mediate a wide range of functionally diverse proteins through protein–protein interactions that are typically phosphorylation-dependent. However, the interactions between 14-3-3 proteins and the major regulators of nature rubber (NR) biosynthesis in H. brasiliensis have not been fully elucidated. In this study, we obtained 81 essential client proteins that interacted with H. brasiliensis 14-3-3 proteins (HbGF14s) through yeast two-hybrid (Y2H) screening. These interaction partners were involved in plant signal transduction, metabolism, development, and NR biosynthesis including small rubber particle protein (SRPP), rubber elongation factor (REF), and MYC2, etc. Furthermore, the interaction of HbGF14c and HbSRPP of H. brasiliensis was confirmed in plants through bimolecular fluorescence complementation (BiFC) assays and in vitro with Pull-down assays. Specifically, the RVSSYLP motif was found to mediate the interaction between HbSRPP and HbGF14c. The findings of this study provide a theoretical basis for the elucidation of the molecular regulation mechanism of the 14-3-3 proteins involved in NR biosynthesis, which could be used to enhance the production of rubber trees through genetic improvement.

The rubber elongation factor (REF) is the most abundant protein in the latex of Hevea brasiliensis, which is closely related to natural rubber biosynthesis. In order to gain a deeper understanding of the transcriptional regulation mechanism of HbREF1, a 1758 bp genomic DNA fragment of the HbREF1 promoter was isolated. Promoter sequence analysis revealed several transcription factor binding sites in the HbREF1 promoter, such as bZIP, bHLH, EIL, AP2/ERF, MYB, and Trihelix. To assess the promoter activity, a series of HbREF1 promoter deletion derivatives were created and fused with firefly luciferase (LUC). The LUC image demonstrated that all of the HbREF1 promoters exhibited transcriptional activity. Furthermore, the assay revealed the presence of multiple regulatory elements within the promoter region that negatively regulate the transcriptional activity. Subsequent analysis of the transcriptional activity following treatment with phytohormones identified an ABA-responsive element located between −583 bp and −200 bp, an ET-responsive element between −718 bp and −583 bp, a JA-responsive element between −1758 bp and −1300 bp, and a SA-responsive element between −1300 bp and −718 bp. These results were largely consistent with the predictions of cis-acting elements. This study has established significant groundwork for future investigations into the regulatory mechanism of HbREF1.