The DEAD-box family has been shown to play an important role in a variety of abiotic stresses, but little is known in studies of mangrove plants. Here, the effects of cold stress on various physiological changes and the role of the DEAD-box RNA helicase family in response to cold stress were determined. First, we identified 73 DEAD-box RNA helicase family members in L. littorea. Second, the evolutionary relationships between the DEAD family in L. littorea and the model species Arabidopsis thaliana were investigated by evolutionary phylogenetic analysis. Finally, qRT-PCR study of representative DEAD-box genes showed that DEAD-box genes played a major role in the low-temperature stress response of L. littorea. Furthermore, we found that LlDEAD48, LlDEAD36, and LlDEAD47 might be involved in the maintenance of chlorophyll function, and LlDEAD43 might play a role in the maintenance of mitochondrial function in L. littorea under cold stress.

Lumnitzera littorea (Jack) Voigt is among the most endangered mangrove species in China. The morphology and evolution of L. littorea flowers have received substantial attention for their crucial reproductive functions. However, little is known about the genomic regulation of flower development in L. littorea. In this study, we characterized the morphology of two kinds of L. littorea flowers and performed comparative analyses of transcriptome profiles of the two different flowers. Morphological observation showed that some flowers have a column embedded in the petals while others produce a stretched flower style during petal unfolding in flowering. By using RNA-seq, we obtained 138,857 transcripts that were assembled into 82,833 unigenes with a mean length of 1055.48 bp. 82,834 and 34,997 unigenes were assigned to 52 gene ontology (GO) functional groups and 364 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. A total of 4,267 differentially expressed genes (DEGs), including 1,794 transcription factors (TFs), were identified between two types of flowers. These TFs are mainly involved in bHLH, B3, bZIP, MYB-related, and NAC family members. We further validated that 12 MADS-box genes, including 4 MIKC-type and 8 M-type TFs, were associated with the pollinate of L. littorea by herkogamy. Our current results provide valuable information for genetic analysis of L. littorea flowering and may be useful for illuminating its adaptive evolutionary mechanisms.

Plant basic helix-loop-helix (bHLH) transcription factors play pivotal roles in responding to stress, including cold and drought. However, it remains unclear how bHLH family genes respond to these stresses in Kandelia obovata. In this study, we identified 75 bHLH members in K. obovata, classified into 11 subfamilies and unevenly distributed across its 18 chromosomes. Collineation analysis revealed that segmental duplication primarily drove the expansion of KobHLH genes. The KobHLH promoters were enriched with elements associated with light response. Through RNA-seq, we identified several cold/drought-associated KobHLH genes. This correlated with decreased net photosynthetic rates (P_n) in the leaves of cold/drought-treated plants. Weighted gene co-expression network analysis (WGCNA) confirmed that 11 KobHLH genes were closely linked to photoinhibition in photosystem II (PS II). Among them, four Phytochrome Interacting Factors (PIFs) involved in chlorophyll metabolism were significantly down-regulated. Subcellular localization showed that KobHLH52 and KobHLH30 were located in the nucleus. Overall, we have comprehensively analyzed the KobHLH family and identified several members associated with photoinhibition under cold or drought stress, which may be helpfulfor further cold/drought-tolerance enhancement and molecular breeding through genetic engineering in K. obovata.

Endogenous melatonin (MEL) and abscisic acid (ABA) are involved in the adaptation of plants to environmental stresses. The application of exogenous serotonin (SER) to plants can enhance their tolerance to abiotic stress, such as cold. However, the mechanism associated with serotonin-mediated defense against cold-induced damage in mangroves is still poorly understood. In this study, we demonstrated that mangrove (Kandelia obovata) seedlings sprayed with 200 μmol·L⁻¹ serotonin exhibited enhanced cold tolerance, as shown by reduced damage to leaves and loss of photosynthesis when exposed to low-temperature conditions. The mechanism associated with the cold adaptation of K. obovata seedlings upon treatment with serotonin was subsequently investigated by transcriptomic analysis. Serotonin treatment caused changes in differentially expressed genes (DEGs) involved in the regulation of melatonin (MEL) and ABA biosynthesis and defense responses against cold stress. Under low-temperature stress, serotonin-treated seedlings showed a significant increase in the endogenous levels of melatonin and ABA. By contrast, under normal growth conditions, K. obovata seedlings treated with serotonin displayed no substantial change in melatonin level, whereas ABA level significantly increased. These findings demonstrated that serotonin treatment might play an important role in the enhanced resistance to cold in K. obovata and that such an effect would depend on the activation of endogenous melatonin and ABA synthesis.