Coconut palms (Cocos nucifera L.) are globally significant palms with both economic and cultural value. Despite the increasing demand for coconut products, production is decreasing globally due to palm senility, pests, and diseases. It has been estimated that over half of the world’s coconut palms need to be replaced immediately. The coconut industry has acknowledged that conventional propagation methods are unlikely to yield sufficient high-quality planting material. Therefore, coconut tissue culture is considered a potential solution to this problem. By using coconut tissue culture, a large number of plantlets can be obtained in a short period of time. In this study, the quality of explants and the development stage (visible shoot/non-visible shoot) of coconut used for micropropagation were examined. To our knowledge, little research has been undertaken on this aspect of coconut micropropagation. Our results indicated that tender coconut fruit exhibited an advantage over mature fruits. In addition, coconut plumule explants subjected to an extended storage of 15 days demonstrated enhanced development compared to those without storage. Notably, smaller embryos utilized as explants displayed superior callus formation compared to their larger counterparts. Finally, embryos possessing shoots exhibited improved callus initiation, albeit accompanied by a more pronounced browning effect. Further investigations are required to obtain more knowledge about the most suitable conditions for plumule explants that lead to optimal callus initiation.

As one of the most important members of the palm family, coconut (Cocos nucifera L.) currently faces a substantial gap between demand and production. Current plantings of this crop are aging, and these traditional varieties are susceptible to several devastating pests and diseases. Consequently, there is an urgent need to replant and expand coconut lands with new, genetically superior varieties. Such replanting cannot be met through the conventional method of seed nut planting, and tissue culture has emerged as a likely solution to address this problem. However, due to certain technical barriers, elevated costs, and a need for improved efficiency, the development of automated and highly efficient tissue culture techniques is yet to be developed. The present research explores the potential of an in vitro temporary immersion system (TIS) to improve the production of somatic embryogenic callus for plantlet regeneration. Results indicated that, in comparison to the conventional agar-based method used to produce coconut somatic embryogenic callus, the TIS method significantly enhanced embryogenic callus production. The optimal biomass of callus for inoculating the TIS was determined to be 0.2 g in each 900 mL vessel and the most favorable embryogenic developmental stage for employing TIS was the globular stage of embryo development. The most effective immersion time to give the highest yield of embryogenic callus was 5 min every 6 h. This foundational research demonstrates that a TIS step is likely to be important to rapidly produce, on a large scale, coconut plantlets to meet the escalating demand for materials for the replanting of coconut lands.

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The coconut (Cocos nucifera L.), a valuable tropical crop, is rapidly declining in genetic diversity due to natural disasters, pest and disease attack, and land clearing for other crops. Seed banking is impractical for coconut conservation due to its large, recalcitrant seed, and maintaining field gene bank collections is costly and vulnerable to environmental pressures. Cryopreservation offers a promising alternative method for conserving coconut genetic diversity, but the success in recovering cryopreserved materials remains limited, with few studies consistently reporting high rates of recovery. This highlights the need for improved cryopreservation protocols, particularly in tissue dehydration, which is one of the critical steps in the process of cryopreservation and plant recovery. A desiccator was developed that enabled rapid embryo dehydration with ultra-dry airflow. The desiccator reduced embryo moisture content to 20% (the predetermined viability threshold) within 6 h representing a 2-h improvement when compared to a previous dehydration approach, while maintaining a high germination rate (71%). Smaller embryos (500 to 550 mg fresh weight) desiccated faster than larger embryos (800 to 900 mg fresh weight) but germination was reduced (30%), making small embryos unsuitable for cryopreservation. A 5-day sucrose (0.4 M) pre-treatment further reduced the dehydration time to 4 h, while maintaining a high germination rate (70%). These advances in the use of a sucrose pre-treatment, the rapid embryo dehydration, and selection of large embryos size will help to enhance the success of coconut embryo cryopreservation and recovery.

Sugar-Will-Eventually-be-Exported Transporters (SWEETs) play a crucial role in sugar transport in plants, mediating both plant development and stress responses. Despite their importance, there has been limited research characterizing the functional characteristics of CnSWEET genes in coconut (Cocos nucifera). In this study, we conducted a systematic analysis of SWEET genes in coconut using bioinformatics, subcellular localization studies, in silico promoter analysis, and functional assays with yeast mutants. A total of 16 CnSWEET genes were identified and grouped into four clades. Clade I contained the highest number of genes (eight), derived from four pairs of duplicated genomic segments. In contrast, the other clades had fewer genes (one to four) compared to those in Arabidopsis and other species in the Arecaceae family. An extensive analysis of gene expansion using MSCanX indicated significant divergence in gene expansion patterns, both between and within monocots and dicots, as well as among closely related species within the same family. Notable variations in conserved protein motifs and the number of transmembrane helices (TMHs) were detected within Clade I compared to other clades, affecting the subcellular localization of CnSWEET proteins. Specifically, seven TMHs were associated with proteins located in the cell membrane, while CnSWEET2A, which had five TMHs, was found in both the cell membrane and cytosol. Promoter analysis revealed that some CnSWEET genes contained endosperm or seed specific motifs associated with specific endosperm expression, consistent with expression patterns observed in transcriptome data. Functional analysis of five CnSWEET genes, with transcript sequences supported by transcriptome data, was conducted using yeast mutant complementation assays. This analysis demonstrated diverse transport activities for sucrose, fructose, glucose, galactose, and mannose. Our findings provide valuable insights into the functional diversity of SWEET genes in coconuts and their potential roles in stress responses and plant development.