The coconut is an important tropical economical crop and exhibits high tolerance to various types of salinity stress. However, little is known about the molecular mechanism underlying its salt tolerance. In this study, RNA-Seq was applied to examine the different genes expressed in four coconut varieties when exposed to a salt environment, resulting in the generation of data for 48 transcriptomes. Comparative transcriptome analysis showed that some genes involved in cutin and wax biosynthesis were significantly upregulated in salt treatment compared to the control, including CYP86A4, HTH, CER1, CER2, CER3, DCR, GPAT4, LTP3, LTP4, and LTP5. In particular, the expression of CER2 was induced more than sixfold, with an RPKM value of up to 205 ten days after salt treatment in Hainan Tall coconut, demonstrating superior capacity in salt tolerance compared to dwarf coconut varieties. However, for yellow dwarf and red dwarf coconut varieties, the expression level of the CER2 gene was low at four different time points after exposure to salt treatment, suggesting that this gene may contribute to the divergence in salt tolerance between tall and dwarf coconut varieties. Cytological evidence showed a higher abundance of cuticle accumulation in tall coconut and severe damage to cuticular wax in dwarf coconut.

Reducing the time before the flowering stage in coconut (Cocos nucifera L.) trees greatly influences yield, yet the mechanisms driving the switch from vegetative to reproductive growth are not well understood, especially the role of phosphorus in this transition. In this study, dwarf coconut plants of the same cultivation age were selected and categorized into the vegetative phase (VP) or the reproductive phase (RP). By examining the phenotypic traits, nutrient variations in the roots and soil, and the transcriptional expression of relevant genes in the roots across both phases, we investigated the potential mechanisms driving the transition from the VP to the RP in coconuts. The shoots of coconuts in the RP were significantly taller compared to those in the VP. Moreover, the phosphorus concentration in the roots of coconuts during the RP was 1.31 times higher than in the VP, which may be linked to the significant upregulation of the PT1 genes AZ11G0219160 and AZ02G0034860 in the roots of coconuts in the RP. In addition, all phosphorus-containing metabolites in the roots during the RP showed a significant increase, particularly those related to long-chain fatty acids and ribonucleotide metabolites. This suggests that coconut roots may facilitate the progression from vegetative to reproductive growth by enhancing phosphorus uptake via PT1s and promoting the synthesis and accumulation of phosphorus-containing metabolites.

Aromatic coconut (Cocos nucifera L.) is valued in the consumer market primarily for its distinctive fragrance, which is largely attributed to the compound 2-acetyl-1-pyrroline (2AP). The accumulation of 2AP has been observed in several crops, such as rice, when exposed to salt stress. In rice, exposure to salt stress influences the activity of enzymes, alters amino acid metabolism, and modulates the expression of genes associated with 2AP formation. Nevertheless, the processes responsible for 2AP biosynthesis in aromatic coconut under salt stress conditions are still not well clarified. In this study, five-month-old aromatic coconut seedlings were subjected to four distinct levels of sodium chloride (NaCl) treatment (0, 100, 200, and 300 mM). This experiment was conducted to investigate the mechanisms involved in salt-induced responses and the biosynthesis of 2AP in aromatic coconut. Although salt stress did not produce any apparent injury in the coconut seedlings, it led to a marked decline in chlorophyll content. Meanwhile, salt stress markedly enhanced the accumulation of betaine and boosted the activities of antioxidant enzymes such as superoxide dismutase and catalase. The aromatic coconut demonstrated a moderate level of salt tolerance. Salt stress also had a significant influence on 2AP biosynthesis. Under salt stress conditions, the 2AP content increased substantially, reaching its highest level with a 93.55% rise compared to the control. Furthermore, the synthesis of 2AP in aromatic coconut under salt stress appears to be primarily regulated through the metabolic pathways of proline and glutamate. Therefore, salt stress enhances 2AP production, with 200 mM NaCl identified as the optimal concentration for its accumulation.

The distinct aroma of the coconut (Cocos nucifera L.) is attributed to the presence of 2-acetyl-1-pyrroline (2AP), although the molecular and genetic pathway associated with its stress tolerance and aroma properties is largely unexplored. The glutamate decarboxylase (GAD) enzyme is involved in the formation of γ-aminobutyric acid (GABA), which is implicated in both adaptation to abiotic stress and 2AP synthesis. In this study, we comprehensively characterized the GAD gene family in coconuts and related Arecaceae species, revealing three highly conserved paralogs that diverged before speciation. It is established that CnGAD2 harbors a typical structure for the glutamate decarboxylase domain with palm-specific long-intron structure and the crucial C-terminal residues required for CaM binding. Notably, CnGAD2 was highly expressed in the middle leaves of aromatic germplasm compared with non-aromatic germplasm. Its expression level was also strongly associated with salt-stress severity and the accumulation of GABA and 2AP. An allele-based haplotype association study revealed a dominant haplotype that has higher levels of 2AP content than other haplotypes. Collectively, these findings suggest that CnGAD2 may function as an important link between salt-stress signaling and aroma biosynthesis. The identified dominant haplotype could therefore serve as a promising genetic resource for improving stress tolerance and flavor quality in coconut, an economically important tropical crop.

Aromatic coconut is a special variety of coconut. Its unique “pandan-like” aroma has won it great popularity among consumers, endowing it with considerable market potential. In our previous study, 2-acetyl-1-pyrroline (2AP), which serves as the main source of the “pandan-like” aroma in aromatic coconut, was found to exhibit significant variation among distinct aromatic coconut individuals. Now, the regulatory mechanism of 2AP has been clarified in fragrant rice, and the ProDH gene is the key gene for 2AP regulation. To further understand the regulation mechanism of 2AP content in aromatic coconut, we cloned and identified the CnProDH gene, the key gene of 2AP regulation in aromatic coconut. The results showed that the CnProDH gene had the typical ProDH structural domain, and its full-length sequence is 23,667 bp, containing 5 exons and a coding sequence (CDS) of 1599 bp. The CnProDH gene encodes a protein that possesses a β₈α₈ barrel structure, consisting of 532 amino acids (aa), with a molecular mass of 58,076.63 kDa and an isoelectric point of 7.11. To further understand the regulatory mechanism of CnProDH in aromatic coconut, we also constructed a yeast one-hybrid (Y1H) library for aromatic coconut. Through the Y1H experiment, combined with the prediction and analysis of cis-acting elements in the promoter of the CnProDH gene, three possible regulatory factors, including CnYABBY2, CnSAP8, and CnBRD3, were identified. These findings provide a molecular basis for clarifying and solving the problem of variations in 2AP content across different aromatic coconuts.