Xinkai Cai

Late Embryogenesis Abundant (LEA) proteins play vital roles in plant responses to abiotic stress and developmental regulation. Pineapple (Ananas comosus L.) is a major tropical fruit crop with high economic value, but its production is often threatened by cold stress, particularly in regions at the northern margin of its cultivation. Despite the recognized importance of LEA proteins in stress adaptation, their genomic landscape and functional characteristics in pineapple remain largely unexplored. In this study, 37 AcLEA genes were identified in the pineapple (Ananas comosus L.) genome and classified into six subfamilies, with LEA_2 being the largest. Most AcLEA proteins were predicted to be hydrophilic, thermally stable, and intrinsically disordered, consistent with typical LEA protein characteristics. Phylogenetic and collinearity analyses revealed species-specific expansion patterns, primarily driven by segmental duplication events. Most duplicated gene pairs shared similar exon–intron structures, motif compositions, and expression profiles, although several displayed signs of functional divergence based on distinct expression patterns, Ka/Ks ratios > 1, and motif differences. Promoter cis-element, transcription factor, and miRNA network predictions indicated that AcLEA genes are widely involved in stress responses as well as growth and development. Expression profiling showed that many AcLEA genes including AcLEA32, AcLEA7, AcLEA9, AcLEA30, AcLEA29, AcLEA33, and AcLEA18 were significantly upregulated under cold stress and declined upon stress removal, indicating a potential role in cold tolerance. Some AcLEA genes, such as AcLEA32 and AcLEA33, showed faster and stronger induction under cold stress in the cold-tolerant cultivar “Comte de Paris” (BL) compared to the sensitive cultivar “Tainong No. 20” (NN), suggesting that differential gene responsiveness may contribute to cultivar-specific cold tolerance. Additionally, most AcLEA genes exhibited distinct spatiotemporal expression patterns across floral organs and fruit at various developmental stages, suggesting their involvement in reproductive development. These findings provide a foundation for future functional studies and highlight candidate genes for improving cold resilience and developmental traits in pineapple through molecular breeding.

Understanding the metabolic characteristics of pineapple varieties is crucial for market expansion and diversity. This study performed comparative metabolomic analysis on the “Comte de Paris” (BL) and three Taiwan-introduced varieties: “Tainong No. 11” (XS), “Tainong No. 23” (MG), and “Tainong No. 13” (DM). A total of 551 metabolites were identified across the four varieties, with 231 metabolites exhibiting no significant differences between all varieties. This included major sugars such as sucrose, glucose, and fructose, as well as key acids like citric, malic, and quinic acids, indicating that the in-season maturing fruits of different pineapple varieties can all achieve good sugar–acid accumulation under suitable conditions. The differentially accumulated metabolites (DAMs) that were identified among the four varieties all primarily belonged to several major subclasses, including phenolic acids, flavonoids, amino acids and derivatives, and alkaloids, but the preferentially accumulated metabolites in each variety varied greatly. Specifically, branched-chain amino acids (L-leucine, L-isoleucine, and L-valine) and many DAMs in the flavonoid, phenolic acid, lignan, and coumarin categories were most abundant in MG, which might contribute to its distinct and enriched flavor and nutritional value. XS, meanwhile, exhibited a notable accumulation of aromatic amino acids (L-phenylalanine, L-tryptophan), various phenolic acids, and many lignans and coumarins, which may be related to its unique flavor profile. In DM, the dominant accumulation of jasmonic acid might contribute to its greater adaptability to low temperatures during autumn and winter, allowing off-season fruits to maintain good quality. The main cultivar BL exhibited the highest accumulation of L-ascorbic acid and many relatively abundant flavonoids, making it a good choice for antioxidant benefits. These findings offer valuable insights for promoting different varieties and advancing metabolome-based pineapple improvement programs.

Key message Twenty-nine GRAS genes were identified in passion fruit, the N-terminal regions and 3D (three-dimensional) structures were closely related with their tissue-specific expression patterns. Candidate PeGRASs for enhancing stress resistance were identified. Abstract Passion fruit (Passiflora edulis Sims) is a tropical fruit crop with significant edible and ornamental value, but its growth and development are highly sensitive to environmental conditions. The plant-specific GRAS gene family plays critical roles in regulating growth, development, and stress responses. Here, we performed the first comprehensive analysis of the GRAS gene family in passion fruit. A total of 29 GRAS genes were identified and named PeGRAS1 to PeGRAS29 based on their chromosomal locations. Phylogenetic analysis using GRAS proteins from passion fruit, Arabidopsis, and rice revealed that PeGRAS proteins could be classified into 10 subfamilies. Compared to Arabidopsis, passion fruit lacked members from the LAS subfamily but gained one GRAS member (PeGRAS9) clustered with the rice-specific Os4 subfamily. Structural analysis performed in silico revealed that most PeGRAS members were intron less and exhibited conserved motif patterns near the C-terminus, while the N-terminal regions varied in sequence length and composition. Members within certain subfamilies including DLT, PAT1, and LISCL with similar unstructured N-terminal regions and 3D structures, exhibited similar tissue-specific expression patterns. While PeGRAS members with difference in these structural features, even within the same subfamily (e.g., DELLA), displayed distinct expression patterns. These findings highlighted that the N-terminal regions of GRAS proteins may be critical for their specific functions. Moreover, many PeGRAS members, particularly those from the PAT1 subfamily, were widely involved in stress responses, with PeGRAS19 and PeGRAS26 likely playing roles in cold tolerance, and PeGRAS25 and PeGRAS28 in drought resistance. This study provides a foundation for further functional research on PeGRASs and offers potential candidates for molecular breeding aimed at enhancing stress tolerance in passion fruit.

Pineapple (Ananas comosus L.) is a major tropical fruit crop with considerable economic importance, and its growth and development are significantly impacted by low temperatures. The plant-specific GRAS gene family plays crucial roles in diverse processes, including flower and fruit development, as well as in stress responses. However, the role of the GRAS family in pineapple has not yet been systematically analyzed. In this study, 43 AcGRAS genes were identified in the pineapple genome; these genes were distributed unevenly across 19 chromosomes and 6 scaffolds and were designated as AcGRAS01 to AcGRAS43 based on their chromosomal locations. Phylogenetic analysis classified these genes into 14 subfamilies: OS19, HAM-1, HAM-2, SCL4/7, LISCL, SHR, PAT1, DLT, LAS, SCR, SCL3, OS43, OS4, and DELLA. Gene structure analysis revealed that 60.5% of the AcGRAS genes lacked introns. Expression profiling demonstrated tissue-specific expression, with most AcGRAS genes predominantly expressed in specific floral organs, fruit tissues, or during particular developmental stages, suggesting functional diversity in pineapple development. Furthermore, the majority of AcGRAS genes were induced by cold stress, but different members seemed to play distinct roles in short-term or long-term cold adaptation in pineapple. Notably, most members of the PAT1 subfamily were preferentially expressed during late petal development and were upregulated under cold stress, suggesting their special roles in petal development and the cold response. In contrast, no consistent expression patterns were observed among genes in other subfamilies, suggesting that various regulatory factors, such as miRNAs, transcription factors, and cis-regulatory elements, may contribute to the diverse functions of AcGRAS members, even within the same subfamily. This study provides the first comprehensive analysis of GRAS genes in pineapple, offers valuable insights for further functional investigations of AcGRASs and provides clues for improving pineapple cold resistance breeding.

Passion fruit, an economically valuable fruit crop, is highly vulnerable to adverse climate conditions. The HVA22 genes, recognized as abscisic acid (ABA) and stress-inducible, play vital roles in stress response and growth regulation in diverse eukaryotic organisms. Here, six HVA22 genes were firstly identified in passion fruit genome and all predicted to be localized within the endoplasmic reticulum. Phylogenetic analyses showed that all PeHVA22s were divided into four subgroups. The gene structural features of PeHVA22 genes clustered in the same subgroup were relatively conserved, while the gene structure characteristics of PeHVA22s from different subgroups varied significantly. PeHVA22A and PeHVA22C closely clustered with barley HVA22 in Group II, were also induced by ABA and drought stress treatment, suggesting conserved roles similar to barley HVA22. Meanwhile, most PeHVA22s exhibited induced expression post-drought treatment but were suppressed under salt, low and high-temperature conditions, indicating a unique role in drought response. Additionally, PeHVA22s displayed tissue-specific expression patterns across diverse tissues, except for PeHVA22B which maybe a pseudogene. Notably, PeHVA22C, PeHVA22E, and PeHVA22F predominantly expressed in fruit, indicating their involvement in fruit development. Almost all PeHVA22s showed variable expression at different developmental stages of stamens or ovules, implying their roles in passion fruit’s sexual reproduction. The intricate roles of PeHVA22s may result from diverse regulatory factors including transcription factors and CREs related to plant growth and development, hormone and stress responsiveness. These observations highlighted that PeHVA22s might play conserved roles in ABA response and drought stress tolerance, and also be participated in the regulation of passion fruit growth and floral development.

Passion fruit (Passiflora edulis) is an economically valuable tropical fruit crop renowned for its nutritious juice, aromatic fragrance, and vibrant flowers with distinct floral structure, known as corona. These unique floral features make passion fruit a good candidate for the study of floral organogenesis. MADS-box genes play essential functions in various aspects of plant growth and development, especially for floral morphogenesis. However, a comprehensive investigation of MADS-box gene family members in passion fruit has not yet been conducted. Here, 52 MADS-box genes were identified in the passion fruit genome and classified into two types with five subgroups (type I: Mα, Mβ, Mγ; type II: MIKC^C, MIKC*) based on phylogeny. The notable reduction in the abundance of type I MADS-box genes within the passion fruit genome, in contrast to Arabidopsis, is thought to be a contributing factor to the diminished gene number within this gene family. The structural analysis illustrated that PeMADS proteins within the same subfamily are relatively conserved. Moreover, we proposed the putative flower regulation ABC(D)E model of passion fruit and explored the regulatory genes of the characteristic structure corona. We found that the regulation of petals and stamens in passion fruit is similar to that of Arabidopsis, but differs in terms of carpels and sepals. The distinct coronas were predominately controlled by B-, C(D)- and E-class genes, supporting the speculation that the corona might have originated from stamens. Except for the core functions in the floral meristem, many PeMADS genes were also involved in the development regulation of non-floral tissues and/or in phytohormone and stress responses. The co-presence of diverse cis-regulatory elements associated with growth/developmental regulation, phytohormone and stress responses in the promoter regions of PeMADSs might be closely related to their diverse regulatory roles. The results of this study provide valuable insights into the MADS-box genes in passion fruit and their involvement in the development of floral structure. These discoveries lay the foundation for the cultivation of exceptional ornamental varieties of passion fruit.

Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.) stands as one of the pivotal afforestation tree species and timber resources in southern China. Nevertheless, the occurrence of seed abortion and a notably high proportion of astringent seeds significantly curtail the yield and quality of elite seeds, resulting in substantial economic losses. The development of astringent seeds is accompanied by significant physiological and biochemical alterations. Here, the first combined lipidomic and metabolomic analysis was performed to gain a comprehensive understanding of astringent seed traits. A total of 744 metabolites and 616 lipids were detected, of which 489 differential metabolites and 101 differential lipids were identified. In astringent seeds, most flavonoids and tannins, as well as proline and γ-aminobutyric acid, were more accumulated, along with a notable decrease in lipid unsaturation, indicating oxidative stress in the cells of astringent seeds. Conversely, numerous elemental metabolites were less accumulated, including amino acids and their derivatives, saccharides and alcohols, organic acids and nucleotides and their derivatives. Meanwhile, most lipid subclasses, mainly associated with energy storage (triglyceride and diglyceride) and cell membrane composition (phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine), also exhibited significant reductions. These results reflected a disruption in the cellular system or the occurrence of cell death, causing a reduction in viable cells within astringent seeds. Furthermore, only one lipid subclass, sphingosine phosphate (SoP), was more accumulated in astringent seeds. Additionally, lower accumulation of indole-3-acetic acid and more accumulation of salicylic acid (SA) were also identified in astringent seeds. Both SA and SoP were closely associated with the promotion of programmed cell death in astringent seeds. Collectively, our study revealed significant abnormal changes in phytohormones, lipids and various metabolites in astringent seeds, allowing us to propose a model for the development of astringent seeds in Chinese fir based on existing research and our findings. This work enriches our comprehension of astringent seeds and presents valuable bioindicators for the identification of astringent seeds.