Search Results (32)

The Lateral Organ Boundaries Domain (LBD) gene family encodes plant-specific transcription factors that play pivotal roles in growth, development, and stress responses. However, a comprehensive genome-wide analysis of the LBD family in watermelon (Citrullus lanatus) has not been conducted until now. In this study, we identified 39 ClLBD genes using the latest watermelon reference genome and systematically analyzed the function of ClLBD2 in root development. These ClLBDs are unevenly distributed across 10 chromosomes except Chr4. Evolutionary analysis grouped the gene family members into six subgroups: Class I (a–e) and Class II. Physicochemical properties and gene structure analysis showed that the ClLBD protein members are tightly conserved. In the promoter regions of ClLBD genes, we identified abundant cis-acting regulatory elements related to abiotic stress and hormone responses. Through RNA-seq analysis from a cucurbit database, we found that several ClLBD genes showed high relative expression in roots, with ClLBD2 being the most highly expressed. Since its subfamily includes AtLBD25, a known root development-related gene, we hypothesized that ClLBD2 might be involved in root development. To validate this, ClLBD2-edited roots were generated using the CRISPR-Cas9 system and Agrobacterium rhizogenes-mediated transformation. Compared to the wild type, the ClLBD2 edited roots exhibited significant reduction in taproot length and lateral root numbers, indicating that ClLBD2 may regulate root development. This study provides the first comprehensive analysis of the LBD gene family in watermelon, offering valuable insights for evolutionary and further functional studies of ClLBD genes. Full article

Fusarium oxysporum f. sp. niveum is an increasingly threatening fungal pathogen that systemically colonizes watermelon plants and severely compromises their productivity by causing destructive vascular wilt disease. While its nonribosomal peptide synthetase NPS6 has been identified as a key virulence factor, the regulatory mechanisms through which it controls downstream gene networks to cause disease remain unclear. To elucidate this regulatory pathway, we constructed a ΔFW-NPS6 knockout mutant and conducted a comparative genome-wide analysis using RNA sequencing, with the wild-type strain as a control. The results revealed 66 NPS6-dependent differentially expressed genes, which were primarily associated with secondary-metabolite biosynthesis (e.g., genes encoding nonribosomal peptide synthetases like NPS2) and pathogen–host interactions (e.g., components of the MAPK signaling pathway), and were enriched in key pathogenic pathways. This finding reveals the virulence regulatory network mediated by NPS6, providing a direct theoretical foundation and crucial molecular targets for developing novel control strategies, such as targeted fungicides or genetic interventions, against Fusarium wilt in watermelon by highlighting NPS6 itself as a potential fungicide target and its downstream pathways (e.g., siderophore biosynthesis) as points for intervention. Full article

Powdery mildew poses a significant threat to watermelon production. The development of disease-resistant varieties through gene editing represents a major focus in current breeding research. In this study, we identified an MLO family gene in watermelon, denoted by ClMLO5b, which is phylogenetically closely related to cucumber CsaMLO8 and melon CmMLO5. Homology modeling revealed high conservation of the three-dimensional protein structures among these orthologs. Expression analysis demonstrated that ClMLO5b is significantly up-regulated upon powdery mildew infection, and the protein localizes to the plasma membrane. To validate its function, we first employed an Agrobacterium rhizogenes-mediated hairy root transformation system to rapidly verify the editing efficiency of two CRISPR/Cas9 targets designed for ClMLO5b. Subsequently, stable transgenic watermelon plants were generated via Agrobacterium tumefaciens-mediated transformation, and a mutant line with homozygous substitutions at target site 2 was obtained. Disease resistance assays showed that, compared to wild-type plants, the Clmlo5b exhibited strongly inhibited mycelial growth, significantly reduced disease severity, and a substantial decrease in spore production after inoculation with powdery mildew. Our findings confirm that ClMLO5b is a key susceptibility gene in watermelon and provide both a promising genetic target and valuable breeding material for developing powdery mildew-resistant watermelon varieties. Full article

Drought stress is a major constraint on watermelon production worldwide. Conventional phenotyping methods for drought tolerance are often low-throughput and fail to capture dynamic physiological responses. This study validated the high-throughput phenotyping platform (Plantarray 3.0) against conventional methods by dynamically evaluating drought tolerance across 30 genetically diverse watermelon accessions. The Plantarray system quantified key dynamic traits, including transpiration rate (TR), transpiration maintenance ratio (TMR), and transpiration recovery ratios (TRRs), revealing distinct drought-response strategies. Principal component analysis (PCA) of these dynamic traits explained 96.4% of the total variance (PC1: 75.5%, PC2: 20.9%), clearly differentiating genotypes. A highly significant correlation (R = 0.941, p < 0.001) was found between the comprehensive drought tolerance rankings derived from Plantarray and conventional phenotyping. We identified five genotypes as highly tolerant and four as highly sensitive. The elite drought-tolerant germplasm, notably the wild species PI 537300 (Citrullus colocynthis) and the cultivated variety G42 (Citrullus lanatus), exhibited superior physiological performance and recovery capacity. The results demonstrate that the Plantarray system not only efficiently screens for drought tolerance but also provides deep insights into dynamic resistance mechanisms, offering a powerful tool and valuable genetic resources for breeding climate-resilient watermelon cultivars. Full article

The regulatory mechanism of brassinolide (BR) signaling in cucurbitaceae crops remains incompletely understood. Previous research demonstrated that the rice genes GW5 and GW5L modulate seed morphology via the BR pathway. However, the conservation of their orthologs in watermelon and their evolutionary trajectory are yet to be elucidated. In this study utilizing the watermelon 97103v2 genome, we identified 15 GW5-LIKE genes. Through structure, phylogenetic tree construction, collinearity, promoter and spatiotemporal expression analysis, we determined that ClGL1 to ClGL3 are the most closely related to GW5 and GW5L. Subsequently, two crucial materials were acquired: the inbred line Jing L6M harboring the homozygous mutant Clgl1, and the near-isogenic line Changhong, a Jing L6M backcross containing the wild-type allele ClGL1. Apart from the disparity in fruit morphology, a clear difference in seed shape was observed between the two. Furthermore, exogenous BR treatment demonstrated that ClGL1 positively regulated the BR signal, aligning with the positive impact of GW5 and GW5L. In conclusion, ClGL1 modulates the morphology of watermelon fruit and seed by enhancing BR signaling, which provides a key gene and theoretical basis for BR signaling evolution and molecular design breeding in Cucurbitaceae. Full article

The genetic diversity of cultivated crops is limited, largely as a result of domestication bottlenecks and the selective pressures imposed during modern breeding. An introgression cross was initiated by mating bitter apple (Citrullus colocynthis), as a wild founder parent, with ‘Charleston Grey’ watermelon (Citrullus lanatus) commercial cultivar, focused on identifying and utilizing trait-enhancing alleles from crop wild relative (CWR). Successful crosses resulted in diverse families, including F₁ hybrids, F₂ population, and backcross (BC) progenies. The study revealed substantial variation among the founder parents and their derived progeny in plant growth and major agronomic fruit traits, highlighting the value of this genetic diversity for breeding programs and demonstrating the potential of Citrullus introgression lines to enhance desired traits in cultivated watermelon. Morphological analysis demonstrated that F₁ progeny resembled the maternal parent for the majority of investigated fruit traits. A considerable proportion of the introgression progeny in the F₂ generation outperformed both parents in total soluble solids and lycopene content, suggesting that crop wild relatives hold strong breeding value through beneficial allelic recombination. BC₁ siblings were closer to the wild watermelon, which is presumably maternally controlled through plastome and mitogenome in crosses between cultivated watermelon and wild bitter apple, which is expected to be retained in successive backcrosses. The study uncovers novel alleles of CWR that preserve extensive genetic variation that is essential for enhancing resilience traits in current breeding lines. These introgression-derived resources provide a critical platform for advancing genetic studies and enhancing crop resilience. Full article

Sebum secreted by sebaceous glands mixes with sweat to form a protective film that aids in maintaining skin health. Reduced sebum production compromises such barrier functions, potentially leading to severe itchiness and inflammation. Therefore, incorporating moisturizers with ingredients promoting sebum secretion is desirable. Wild watermelon possesses moisturizing and antioxidant properties, and its extracts are utilized in skin cosmetics and supplements. This study investigates whether seed watermelon (Citrullus mucosospermus (Fursa))—a species closely related to wild watermelon—influences sebum synthesis and can serve as a skin cosmetic raw ingredient. Several bioactive compounds—including coniferyl alcohol, coniferin, and p-coumaryl alcohol—were identified in the active third fraction of the fruit extract. Subsequently, SZ95 sebocytes stimulated with linoleic acid were stained using Oil Red O to detect lipogenesis facilitated by the identified bioactive compounds. Coniferyl alcohol promoted linoleic acid-stimulated lipogenesis by approximately 2.2-fold at a concentration of 300 µM. Lipidomic analyses confirmed an increase in total lipid content following coniferyl alcohol treatment, with notable increases in cholesterol ester, cardiolipin, and simple lipid content. Overall, these findings suggest that seed watermelon contains compounds that do influence sebum synthesis. Consequently, skin cosmetics containing seed watermelon fruit extracts with linoleic acid may benefit individuals with dry skin. Full article

Members of the heat shock protein 20 (HSP20) family of proteins play an important role in responding to various forms of stress. Here, the expression of ClaHSP17.4 was induced by heat stress in watermelon. Then, a floral dipping approach was used to introduce the pCAMBIA1391b-GFP overexpression vector encoding the heat tolerance-related gene ClaHSP17.4 from watermelon into Arabidopsis thaliana, and we obtained ClaHSP17.4-overexpressing Arabidopsis plants. Under normal conditions, the phenotypes of transgenic and wild-type (WT) Arabidopsis plants were largely similar. Following exposure to heat stress, however, the germination rates (96%) of transgenic Arabidopsis plants at the germination stages were significantly higher than those of wild-type idopsis (17%). Specifically, the malondialdehyde (MDA) content of transgenic Arabidopsis was half that of the control group, while the activities of peroxidase (POD) and superoxide dismutase (SOD) were 1.25 times those of the control group after exposure to high temperatures for 12 h at the seedling stages. The proline content in ClaHSP17.4-overexpressing transgenic Arabidopsis increased by 17% compared to WT plants (* p < 0.05), while the soluble sugar content rose by 37% (* p < 0.05). These results suggest that ClaHSP17.4 overexpression indirectly improves the antioxidant capacity and osmotic regulatory capacity of Arabidopsis seedlings, leading to improved survival and greater heat tolerance. Meanwhile, the results of this study provide a reference for further research on the function of the ClHSP17.4 gene and lay a foundation for breeding heat-tolerant watermelon varieties and advancing our understanding of plant adaptation to environmental stress. Full article

SMALL AUXIN UP-REGULATED RNA (SAURs) genes are acknowledged as auxin-responsive genes that play crucial roles in modulating adaptive growth under abiotic stress conditions. Low temperatures constitute a primary limiting factor that significantly impairs the development, growth, and fruit quality of watermelon plants during the winter and spring seasons. Despite their potential importance, SAURs have not yet been thoroughly investigated or characterized in watermelon. In this study, we identified a positive regulator of the chilling stress response among watermelon SAURs, designated as ClSAUR1. Subcellular localization analysis demonstrated that the protein is directed to both the nucleus and cytoplasm. Quantitative real-time PCR (qRT-PCR) analysis indicated that ClSAUR1 is ubiquitously expressed across various watermelon tissues, with pronounced expression in the roots and leaves. Moreover, qRT-PCR and promoter::β-glucuronidase (GUS) staining assays revealed that the expression of ClSAUR1 is significantly upregulated in response to exogenous abscisic acid (ABA) and chilling stress. The overexpression of ClSAUR1 in tobacco lines was contrasted and analyzed, revealing an increased tolerance to chilling stress. This was evidenced by a reduced degree of wilting and chlorosis compared to wild-type (WT) plants. Furthermore, the overexpressed lines showed reduced reactive oxygen species (ROS) accumulation and increased antioxidant enzyme activity. The qRT-PCR results further indicated that the expression levels of genes associated with abscisic acid (ABA), antioxidant enzymes, and CBF–COR cold-responsive pathways were upregulated in the transgenic tobacco lines. This study provides new insights into the role of ClSAURs in enhancing the cold resistance of watermelon. Full article

Watermelons are one of the most important crop species, and they are enjoyed across the globe; however, the cultivation of watermelon commercial varieties in arid regions is challenging, as they are highly susceptible to water deficit. Conversely, their wild relatives and traditional landraces have shown a higher tolerance to water deficit, which makes them important study material. Therefore, this study was undertaken to evaluate the potential roles of two compatible osmolytes (citrulline and arginine) in the tolerance of local watermelon accessions to drought stress. Four commonly cultivated watermelon accessions were used in this study to evaluate their response when exposed to water deficit stress. The accessions were planted in stress boxes in the greenhouse and allowed to grow until the fourth leaf was fully open and then the water deficit stress was initiated by withholding water for a period of nine days, before rewatering for three days. Data and leaf samples were collected at three-day intervals. The common drought indicators that were assessed, like chlorophyll fluorescence, showed that Clm-08 (wild watermelon) had significantly different results when compared to the other accessions; the Fv/Fm values for days 3, 6, and 9 were significantly higher than those of the other accessions, while phiNPQ was higher in the Clm-08 with average values of 0.41 and 0.41 on days 6 and 9 of the drought stress, respectively. This suggests that the wild watermelon responded differently to drought stress when compared with the other accessions. Arginine and citrulline are important osmolytes that play an important role in stress tolerance, and the results of the current study correlate with the common physiological indicators. The expression pattern for both the biochemical and molecular analyses of the two compatible osmolytes was higher in Clm-08 in comparison with that of the other accessions. The gene expressions of the enzymes in the citrulline and arginine pathways were higher in Clm-08; Cla022915 (CPS) recorded a 6-fold increase on day 6 and Cla002611 (ASS) recorded an 11-fold increase. This suggests that citrulline and arginine play an important role in watermelon tolerance to drought stress. Full article

In 1929, it was reported that yellowing symptoms caused by a tobacco mosaic virus (TMV) yellow mosaic isolate were suppressed in tobacco plants that were systemically infected with a TMV light green isolate. Similar to vaccination, the phenomenon of cross-protection involves a whole plant being infected with an attenuated virus and involves the same or a closely related virus species. Therefore, attenuated viruses function as biological control agents. In Japan, many studies have been performed on cross-protection. For example, the tomato mosaic virus (ToMV)-L₁₁A strain is an attenuated isolate developed by researchers and shows high control efficiency against wild-type ToMV in commercial tomato crops. Recently, an attenuated isolate of zucchini yellow mosaic virus (ZYMV)-2002 was developed and registered as a biological pesticide to control cucumber mosaic disease. In addition, attenuated isolates of pepper mild mottle virus (PMMoV), cucumber mosaic virus (CMV), tobacco mild green mosaic virus (TMGMV), melon yellow spot virus (MYSV), and watermelon mosaic virus (WMV) have been developed in Japan. These attenuated viruses, sometimes called plant vaccines, can be used not only as single vaccines but also as multiple vaccines. In this review, we provide an overview of studies on attenuated plant viruses developed in Japan. We also discuss the application of the attenuated strains, including the production of vaccinated seedlings. Full article

Drought-tolerant plants have become a convenient model to study the mechanisms underlying drought tolerance in order to improve susceptible domesticated relatives. Various studies have shown that local landraces possess superior qualities that help them survive in harsh environmental conditions. One of the key mechanisms that helps with tolerance in crops is timely stomatal regulation. In this study, the physiological, morphological, and molecular stomatal responses in three drought-tolerant landraces (Clm-01–03) and hybrid (Clm-04) watermelons were evaluated under drought stress. The watermelon plants were grown under a water deficit (complete withholding of water) and non-stress conditions. The highest SPAD values were recorded for the Clm-03 and Clm-02 (50 ± 3) watermelon genotypes, and the lowest for Clm-04 (27 ± 0.37), showing this genotype’s tolerance and ability to maintain its systems during drought stress. Fluorescence parameters also gave important clues to the tolerant genotypes of Clm-02 and Clm-03 under drought stress, while the domesticated genotype showed a slow response to fluorescence parameters, which could lead to damage to the photosynthesis apparatus. During the drought period, the wild watermelon was found to have a limited stomatal opening as the drought progressed, and on day 9, it had the smallest opening of 23.1 ± 1.2 µm compared to any other genotype; most importantly, upon re-watering, it showed more rapid recovery than any other genotype. This was also expressed by mRNA quantification of stomatal aperture TFs, with an eight-fold increase in Cla004380 TFs recorded for wild watermelon. All of these mechanisms have been attributed to the tolerance mechanisms of the drought-tolerant watermelon genotype. This study provides important insight into the stomatal responses of probable tolerant watermelon accessions and suggests that improving the stomatal aperture of susceptible domesticated species would also improve their tolerance. Full article

Drought stress has detrimental effects on crop productivity worldwide. A strong root system is crucial for maintaining water and nutrients uptake under drought stress. Wild watermelons possess resilient roots with excellent drought adaptability. However, the genetic factors controlling this trait remain uninvestigated. In this study, we conducted a bulk segregant analysis (BSA) on an F₂ population consisting of two watermelon genotypes, wild and domesticated, which differ in their lateral root development under drought conditions. We identified two quantitative trait loci (qNLR_Dr. Chr01 and qNLR_Dr. Chr02) associated with the lateral root response to drought. Furthermore, we determined that a small region (0.93 Mb in qNLR_Dr. Chr01) is closely linked to drought adaptation through quantitative trait loci (QTL) validation and fine mapping. Transcriptome analysis of the parent roots under drought stress revealed unique effects on numerous genes in the sensitive genotype but not in the tolerant genotype. By integrating BSA, fine mapping, and the transcriptome, we identified six genes, namely L-Ascorbate Oxidase (AO), Cellulose Synthase-Interactive Protein 1 (CSI1), Late Embryogenesis Abundant Protein (LEA), Zinc-Finger Homeodomain Protein 2 (ZHD2), Pericycle Factor Type-A 5 (PFA5), and bZIP transcription factor 53-like (bZIP53-like), that might be involved in the drought adaptation. Our findings provide valuable QTLs and genes for marker-assisted selection in improving water-use efficiency and drought tolerance in watermelon. They also lay the groundwork for the genetic manipulation of drought-adapting genes in watermelon and other Cucurbitacea species. Full article

Acidovorax citrulli can cause bacterial fruit blotch of watermelon, melon, and other cucurbits, and has the potential to cause severe economic losses to growers throughout the world. This article investigated the functions and interactions of the pilR and pilS genes, two important genes in bacterial type IV pili systems, in A. citrulli. For each gene, deletion mutants and complementary strains were constructed via homologous recombination, and their phenotypes were determined. The results showed that the absence of pilR and pilS could significantly reduce the pathogenicity and twitching motility of A. citrulli while increasing the swimming motility, biofilm formation, and in vitro growth. Conversely, complementary strains were no different than the wild-type strain. Using quantitative reverse transcription PCR and promoter activity assays, we confirmed that the deletion of pilR and pilS genes leads to a significant decrease in the transcription level of pilA. Meanwhile, three methods including yeast two-hybrid, glutathione S-transferase pull-down, and luciferase complementation imaging assays were used to verify the direct interaction between the PilR and PilS proteins. These findings revealed the biological function of the pilR and pilS and confirms their regulatory role on pilA. Full article

The auxin/indole-3-acetic acid (Aux/IAA) gene family plays a critical role in auxin-mediated responses and fruit development. However, studies on its role in watermelon are limited. In this study, 29 ClIAA gene members were identified in the watermelon genome and classified into eleven groups. Of note, ClIAA16, which was found to be up-regulated during fruit ripening, was targeted using CRISPR/Cas9 gene editing. Knockout mutants of ClIAA16 exhibited a 3–4 day delay in ripening compared to the wild type, highlighting the regulatory importance of ClIAA16. Our findings shed light on the importance of ClIAA genes in watermelon fruit ripening and pave the way for further functional studies. Full article