Abstract
Changes in plant leaf color during development are directly related to the accumulation or degradation of certain phytochemicals such as anthocyanins. Since some anthocyanins can be beneficial to human health and provide insights into the biology of leaves, the underlying processes and timing by which plants produce these molecules has been the focus of numerous studies. The tree species Hopea hainanensis generally produces green leaves at all growth stages; however, a few explored individuals have been identified possessing red leaves on the top of the seedlings at a young stage. While the phenomenon of leaf color varying with age has been studied in several species, the underlying mechanisms are largely unknown in H. hainanensis. Using a metabolomics approach, the young red leaves in H. hainanensis were found to contain higher levels of anthocyanins and flavonoids than the young green-leaved individuals. Among anthocyanins, pelargonidin and cyanidin were the most likely candidates contributing to the red color of the young leaves. Transcriptome results indicated the genes related to the production of these anthocyanins were significantly upregulated, leading to greater accumulation of red pigments. Specifically, the expression of several MYB and bHLH genes in young red leaf lines was significantly higher than that in the young green leaf lines, especially HhMYB66, HhMYB91, HhMYB6, and HhbHLH70. As such these four transcription factors are probably the main regulatory genes resulting in young red leaves in H. hainanensis. From these results, comparative analyses with other species can be made to better understand the evolution of pigment biosynthesis and how anthocyanins function in plant metabolism and evolution/adaptation.
Highlights
Differences in leaf color have been observed by humans for millennia, especially in temperate climates where many trees reveal colorful pigments in autumn as chlorophyll is degraded and secondary pigments are made evident
To investigate what compounds contribute to the red-colored young leaves in H. hainanensis, the total metabolites of RU, red lines (RL), GU, and green lines (GL) were detected by UPLC-MS/MS followed by hierarchical cluster analysis (HCA) to analyze the differences in accumulated metabolites of the different samples
When the differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) associated with the anthocyanin biosynthesis pathway in H. hainanensis were analyzed separately from the total data set, we found that among the previously screened 30 DEGs, there were 17 genes that had a significant correlation to 3 metabolites, namely dihydroquercetin, dihydrokaempferol, and naringenin-7-o-glucoside, in the comparison of RU vs. GU (Pearson correlation coefficient (PCC) ≥ 0.8)
Summary
Differences in leaf color have been observed by humans for millennia, especially in temperate climates where many trees reveal colorful pigments in autumn as chlorophyll is degraded and secondary pigments are made evident. The current range of this species is restricted to scattered occurrences in dense tropical forests on the island of Hainan and a small number of climactically similar locations in Northern Vietnam. This species is well-known due to the fine-grained durable hardwood it possesses, which has historically been used for a wide variety of applications such as in the production of railroad sleepers, mechanical appliances, fishing vessels, docks, bridges, and in constructing buildings. H. hainanensis is a deep-rooted tree with tolerance to rocky and shallow soils making it highly valuable in ecosystem services related to soil and water preservation and as such it has been often used in landscaping and ecological conservation projects
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