Cucumis Melo Research Articles

Synergistic impact of rotary vacuum evaporator-based conventional and microwave heating treatments on the quality characteristics of Cucumis melo juice

Muskmelon or cantaloupe (Cucumis melo L.) juice receives consumer preference throughout the world due to its sweet taste and flavour. The present research was aimed at investigating the impact of rotary vacuum evaporator-based conventional heating (CH) and microwave heating (MH) on the quality characteristics, i.e., pH, TSS (°Brix), TA (%), conductivity (ms), water evaporation rate (g/min), colour index, and total phenolic content (TPC mg GAE/mL) of Cucumis melo juice. The processing conditions for CH were temperature (50 °C, 70 °C, and 90 °C) and time (1 min, 1.5 min, and 2 min), whereas for MH, the processing conditions were microwave power (450 W, 600 W, and 900 W) and time (1 min, 1.5 min, and 2 min). The results illustrated that CH and MH affected the quality characteristics of Cucumis melo juice significantly. Among all the treatments, the water evaporation rate and TPC were the highest at 900 W for 2 min. Additionally, TCD among all the treatments was found to be minimum (0.56) when Cucumis melo juice was treated at 450 W for 1.5 min. Thus, MH treatment can be considered as an alternative to CH treatment. Overall, the present research provides insight into the treatment process of Cucumis melo juice, which can further enhance its quality characteristics and impact the global market.

First Report of Cucurbit Yellow Vine Disease Caused by Serratia marcescens on Cucurbit Crops in Iowa

Cucurbit yellow vine disease (CYVD) is caused by Serratia marcescens, vectored by squash bugs (Anasa tristis), and is an emerging disease in many parts of the U.S. CYVD can cause 100% yield losses in cucurbits (Bruton et al., 2003). In the summer of 2021, at the Iowa State University Horticultural Research Station (HRS) in Gilbert, Iowa, we observed leaf yellowing, vine decline, and honey-brown discoloration of the phloem of acorn squash (Cucurbita pepo cv. Table Ace) plants in research fields that were infested with squash bugs. In 2022, we observed similar symptoms on pumpkin (Cucurbita maxima cv. Howden) and muskmelon (Cucumis melo cv. Athena) in different fields at the HRS and on giant pumpkins (Cucurbita maxima cv. Prizewinner) in Jones and Ringgold counties. For up to 3 symptomatic plants of each cucurbit species per location, a 20-cm-long stem section immediately above the soil line was excised, surface sterilized by immersion in 10% sodium hypochlorite and 70% ethanol for 2 min each, then triple rinsed in sterile water. The interior of the cross-section tissue was blotted on Luria agar amended with cycloheximide (100 μg/ml) and tetracycline (20 μg/ml) (Stock et al. 2003). Whitish translucent colonies developed after incubation at 28°C for 48 h. The genomic DNA of three isolates from symptomatic plants of muskmelon (MK01), pumpkin (HFP01), and giant pumpkin (AP01), was extracted using the DNeasy Blood and Tissue Kit (Qiagen, Germantown, MD). S. marcescens species-specific primers YV1 (5’-GGGACTTGCTCCCCGG-3’) and YV4 (5’-AACGTCAATTGATGAACGTATTAAGT-3’) (Bruton et al. 2003) were used to amplify part of the 16S rDNA gene, and the primers specific to S. marcescens CYVD strains A79F/A79R (Zhang et al., 2005) were used to amplify part of a major facilitatory superfamily (MFS) transporter gene strain. The sequences of the 16S rRNA PCR product for the three isolates were identical and were deposited in NCBI under Accession OR963533. They shared 100% (395/395 nt) identity with other CYVD strains (Rascoe et al. 2003) and those of other S. marcescens strains in NCBI. The sequences of the amplified region of the MFS transporter gene of the three isolates (NCBI Accession OR962261) were identical and showed a 98.8% (319/323 nt) identity to that of non-CYVD-causing S. marcescens strains, such as N10A28 (Accession CP033623.1). Koch’s postulates were fulfilled by inoculating C. pepo cv. Zephyr plants with either strain HFP01 or phosphate buffer saline (PBS) (10 plants per treatment) 1 wk after seeding by injecting 300 µl of bacteria (~108 CFU/ml) or PBS using a syringe needle. Plants were incubated at 28°C in a growth room for 4 wks. CYVD symptoms similar to those observed in the field developed on 7 out of 10 plants inoculated with strain HFP01 in one study, and 9 out 10 plants in a replicate study, with none of the PBS-inoculated plants showing CYVD symptoms. Bacteria were isolated from the symptomatic plants with selection on tetracycline. The PCR fragments amplified with YV1/YV4 and A79F/A79R were the same size as those of the pre-inoculation strain HFP01. To our knowledge this is the first report of CYVD in Iowa and in the Upper Midwest of the U.S. CYVD is a devastating disease that poses a significant threat to cucurbit production. This report can serve as an alert for the region’s growers and for the development of effective management practices.

MicroRNA Profiling Revealed the Mechanism of Enhanced Cold Resistance by Grafting in Melon (Cucumis melo L.).

Grafting is widely used to improve the resistance to abiotic stresses in cucurbit plants, but the effect and molecular mechanism of grafting on cold stress are still unknown in melon. In this study, phenotypic characteristics, physiological indexes, small-RNA sequencing and expression analyses were performed on grafted plants with pumpkin rootstock (PG) and self-grafted plants (SG) to explore the mechanism of changed cold tolerance by grafting in melon. Compared with SG plants, the cold tolerance was obviously enhanced, the malondialdehyde (MDA) content was significantly decreased and the activities of antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; peroxidase, POD) were significantly increased in PG plants. Depend on differentially expressed miRNA (DEM) identification and expression pattern analyses, cme-miR156b, cme-miR156f and chr07_30026 were thought to play a key role in enhancing low-temperature resistance resulting from grafting. Subsequently, 24, 37 and 17 target genes of cme-miR156b, cme-miR156f and chr07_30026 were respectively predicted, and 21 target genes were co-regulated by cme-miR156b and cme-miR156f. Among these 57 unique target genes, the putative promoter of 13 target genes contained the low-temperature responsive (LTR) cis-acting element. The results of qRT-PCR indicated that six target genes (MELO3C002370, MELO3C009217, MELO3C018972, MELO3C016713, MELO3C012858 and MELO3C000732) displayed the opposite expression pattern to their corresponding miRNAs. Furthermore, MELO3C002370, MELO3C016713 and MELO3C012858 were significantly downregulated in cold-resistant cultivars and upregulated in cold-sensitive varieties after cold stimulus, and they acted as the key negative regulators of low-temperature response in melon. This study revealed three key miRNAs and three putative target genes involved in the cold tolerance of melon and provided a molecular basis underlying how grafting improved the low-temperature resistance of melon plants.

Cuticle properties, wax composition, and crystal morphology of Hami melon cultivars (Cucumis melo L.) with differential resistance to fruit softening

Cuticle wax chemicals are cultivar-dependent and contribute to storage quality. Few research reported on wax analysis between melting flesh-type (MF; ‘Jinhuami 25’) and nonmelting flesh-type (NMF; ‘Xizhoumi 17’ and ‘Chougua’) Hami melons. Chemicals and crystal structures of Hami melon cuticular wax, cell wall metabolism related to fruit melting, and fruit physiology were analyzed to observe wax functions. Results showed that Hami melon cuticle wax predominantly consists of esters, alkanes, alcohols, aldehydes, and terpenoids. MF-type has a lower alkane/terpenoid ratio, concomitant to its higher weight loss and cuticle permeability. Micromorphology of wax crystals appears as numerous platelets with irregular crystals, and the transformation of wax structure in NMF Hami melon is delayed. Waxy components affect cell wall metabolism and physiological quality, which results in the pulp texture difference between MF-type and NMF-type during storage. Results provide a reference for the regulation of wax synthesis in both types of melons.

Post-Harvest Fruit Rot on Muskmelon (Cucumis Melo) Caused by Acremonium Potronii in Iraq

Muskmelon (Cucumis melo) is a vital fruit vegetable thriving in arid and warm climates. Fruit rot is a prevalent issue affecting muskmelon, occurring frequently in fields before harvest and during post-harvest stages. Several pathogenic fungi have been identified as the primary causal agents of fruit rot. In the present study, muskmelon fruits displaying irregular white regions were collected from the Penjwen district, Kurdistan region of Iraq. The isolated causal agent was identified as Acremonium potronii based on both phenotypic characteristics and molecular analysis (ITS and LSU gene sequences). A pathogenicity test was conducted to fulfill Koch’s postulates. This represents the first documented association of Acremonium potronii with muskmelon fruit rot.

Utilizing Xanthan Gum Coatings as Probiotic Bacteria Carriers to Enhance Postharvest Quality and Antioxidants in Fresh-Cut Cantaloupe and Honeydew (Cucumis melo L.) Melons.

Due to spoilage microflora and browning, minimally processed fresh-cut fruits have a short shelf life, and over the years, studies have shown the potential of using edible coatings to extend the shelf life and improve the safety of fresh-cut fruits. Recently, there has been a rise in research on the incorporation of probiotics in edible coatings due to the bespoke health and biopreservation benefits they impart. Therefore, in this study, lactobacillus strains (Lactiplantibacillus plantarum 75 and Bifidobacterium longum) were incorporated into a xanthan edible coating to enhance color retention, sensory properties, antioxidant retention (ascorbic acid, carotenoids, total phenols), and antioxidant activity (FRAP antioxidant power, ABTS scavenger activity) of fresh-cut cantaloupes and honeydew cucumis melo L. melons during cold storage at 5 C and 85% RH for five days. The edible coating was prepared by mixing 0.5% xanthan gum, 1% glycerol, and 2% citric acid solution with L. plantarum 75 (LAB 75) and Bifidobacterium longum bacteria separately, and the final lab count for each strain was made to be 8.0 log CFU/mL. Stable probiotic coatings with ζ-potential of between -39.7 and -51.4 mV and a PdI of 1 were developed, and the incorporation of the probiotic bacteria into the coating was justified using FTIR analysis. The probiotic coatings showed a typical pseudoplastic behavior, in which the viscosity curves fall as the shear rate increases. Thermal stability analysis showed a continuous and multi-step weight reduction in this work, illustrating how the edible coating components interact. The survival of both Lactobacillus strains was recorded on day 5. Both freshly cut melons coated with xanthan and loaded with Lactobacillus strains retained a sufficient quantity of probiotics at the end of storage, while L. plantarum 75 (7 log CFU/g for cantaloupe and 8 log CFU/g for honeydew) retained the highest viability compared to B. longum (6 log CFU/g for cantaloupe and 7 log CFU/g for honeydew). In comparison to the coated and uncoated control samples, the inclusion of L. plantarum 75 in xanthan coatings significantly retained the color properties, pigments (total chlorophyll and carotenoids), ascorbic acid, total phenols, and antioxidant activity (FRAP, DPPH, and ABTS). The overall acceptability of fresh cuts of cantaloupe and honeydew melons coated with xanthan gum loaded with L. plantarum 75 was higher than that of other treatments. Thus, xanthan gum loaded with L. plantarum 75 coating is most suitable for reducing postharvest losses in fresh cuts of honeydew melons and cantaloupe, which will help preserve antioxidant and bioactive properties. The xanthan gum loaded with L. plantarum 75 coatings exhibited the highest preservation impact; therefore, it can be recommended for the fresh-cut industry.

Deciphering the Enhancing Impact of Exogenous Brassinolide on Physiological Indices of Melon Plants under Downy Mildew-Induced Stress.

Melon (Cucumis melo L.) is a valuable horticultural crop of the Cucurbitaceae family. Downy mildew (DM), caused by Pseudoperonospora cubensis, is a significant inhibitor of the production and quality of melon. Brassinolide (BR) is a new type of phytohormone widely used in cultivation for its broad spectrum of resistance- and defense-mechanism-improving activity. In this study, we applied various exogenous treatments (0.5, 1.0, and 2.0 mg·L-1) of BR at four distinct time periods (6 h, 12 h, 24 h, and 48 h) and explored the impact of BR on physiological indices and the genetic regulation of melon seedling leaves infected by downy-mildew-induced stress. It was mainly observed that a 2.0 mg·L-1 BR concentration effectively promoted the enhanced photosynthetic activity of seedling leaves, and quantitative real-time polymerase chain reaction (qRT-PCR) analysis similarly exhibited an upregulated expression of the predicted regulatory genes of photosystem II (PSII) CmHCF136 (MELO3C023596.2) and CmPsbY (MELO3C010708.2), thus indicating the stability of the PSII reaction center. Furthermore, 2.0 mg·L-1 BR resulted in more photosynthetic pigments (nearly three times more than the chlorophyll contents (264.52%)) as compared to the control and other treatment groups and similarly upregulated the expression trend of the predicted key enzyme genes CmLHCP (MELO3C004214.2) and CmCHLP (MELO3C017176.2) involved in chlorophyll biosynthesis. Meanwhile, the maximum contents of soluble sugars and starch (186.95% and 164.28%) were also maintained, which were similarly triggered by the upregulated expression of the predicted genes CmGlgC (MELO3C006552.2), CmSPS (MELO3C020357.2), and CmPEPC (MELO3C018724.2), thereby maintaining osmotic adjustment and efficiency in eliminating reactive oxygen species. Overall, the exogenous 2.0 mg·L-1 BR exhibited maintained antioxidant activities, plastid membranal stability, and malondialdehyde (MDA) content. The chlorophyll fluorescence parameter values of F0 (42.23%) and Fv/Fm (36.67%) were also noticed to be higher; however, nearly three times higher levels of NPQ (375.86%) and Y (NPQ) (287.10%) were observed at 48 h of treatment as compared to all other group treatments. Increased Rubisco activity was also observed (62.89%), which suggested a significant role for elevated carbon fixation and assimilation and the upregulated expression of regulatory genes linked with Rubisco activity and the PSII reaction process. In short, we deduced that the 2.0 mg·L-1 BR application has an enhancing effect on the genetic modulation of physiological indices of melon plants against downy mildew disease stress.

Status, Gaps and Perspectives of Powdery Mildew Resistance Research and Breeding in Cucurbits

Cucurbitaceae, the gourd family of flowering plants, is a very large and diverse family, the order Cucurbitales, contains 95 genera and 950–980 species of food and ornamental plants, and wild and weedy species mostly with high genetic diversity. This review is focused on the most important cucurbit crops (Cucumis sativus, Cucumis melo, Cucurbita spp., Citrullus lanatus, Momordica charantia, Lagenaria siceraria, and Luffa acutangula) and some their wild relatives as hosts of cucurbit powdery mildews (CPM). Powdery mildews (PM) (Ascomycota, Erysiphales) are one of the most frequently encountered and easily visible groups of plant pathogenic fungi with > 900 species. They are obligate biotrophs, they colonize above-ground plant tissues, mostly leaves, though they may also colonize stems, petioles, flowers and fruits, and they are usually debilitators, not killers. These parasitic fungi have been problematic on cucurbits for a long time world-wide, causing serious economic losses in yield and quality. All economically important cucurbit crops host CPM. Seven PM species with different taxonomic positions, host ranges, geographic distributions and ecological requirements are known on cucurbits. CPM species taxonomy and denomination rapidly changed during last few decades through detailed analyses and clarifications. At least three PM species frequently parasitize cucurbits: the endoparasite Leveillula taurica (Lt) with marginal economic importance; and two ectoparasitic species, Golovinomyces orontii (Go), and Podosphaera xanthii (Px), which are economically important world-wide. The two pathogens differ in ecological requirements and distribution, though they may occur together in mixed infections. They are highly variable at the population level for virulence, race identities, and fast adaptation of pathogens. Cucurbit-CPM species interactions are diverse and complicated, and differ between hosts and their respective pathogen genera and species. Here we present a critical overview of obstacles, gaps and recent progress in these matters for six cucurbit genera with respect to resistance resources, genetics of resistance, genetic mapping and development of molecular markers, physiology and mechanisms of resistance, developments in mlo-mediated resistance, patents, and CPM resistance breeding.

Complete genome assembly provides a high-quality skeleton for pan-NLRome construction in melon.

Melon (Cucumis melo L.), being under intensive domestication and selective breeding, displays an abundant phenotypic diversity. Wild germplasm with tolerance to stress represents an untapped genetic resource for discovery of disease-resistance genes. To comprehensively characterize resistance genes in melon, we generate a telomere-to-telomere (T2T) and gap-free genome of wild melon accession PI511890 (C. melo var. chito) with a total length of 375.0 Mb and a contig N50 of 31.24 Mb. The complete genome allows us to dissect genome architecture and identify resistance gene analogs. We construct a pan-NLRome using seven melon genomes, which include 208 variable and 18 core nucleotide-binding leucine-rich repeat receptors (NLRs). Multiple disease-related transcriptome analyses indicate that most up-regulated NLRs induced by pathogens are shell or cloud NLRs. The T2T gap-free assembly and the pan-NLRome not only serve as essential resources for genomic studies and molecular breeding of melon but also provide insights into the genome architecture and NLR diversity.

Comparison of four performance models in quantifying the inequality of leaf and fruit size distribution.

The inequality in leaf and fruit size distribution per plant can be quantified using the Gini index, which is linked to the Lorenz curve depicting the cumulative proportion of leaf (or fruit) size against the cumulative proportion of the number of leaves (or fruits). Prior researches have predominantly employed empirical models-specifically the original performance equation (PE-1) and its generalized counterpart (GPE-1)-to fit rotated and right-shifted Lorenz curves. Notably, another potential performance equation (PE-2), capable of generating similar curves to PE-1, has been overlooked and not systematically compared with PE-1 and GPE-1. Furthermore, PE-2 has been extended into a generalized version (GPE-2). In the present study, we conducted a comparative analysis of these four performance equations, evaluating their applicability in describing Lorenz curves related to plant organ (leaf and fruit) size. Leaf area was measured on 240 culms of dwarf bamboo (Shibataea chinensis Nakai), and fruit volume was measured on 31 field muskmelon plants (Cucumis melo L. var. agrestis Naud.). Across both datasets, the root-mean-square errors of all four performance models were consistently smaller than 0.05. Paired t-tests indicated that GPE-1 exhibited the lowest root-mean-square error and Akaike information criterion value among the four performance equations. However, PE-2 gave the best close-to-linear behavior based on relative curvature measures. This study presents a valuable tool for assessing the inequality of plant organ size distribution.

CPPU Improves Fruit Setting and Growth in Greenhouse-grown Oriental Melons (Cucumis melo L. var. makuwa Makino)

The production of melons (Cucumis melo L.) in greenhouses relies on pollination. Extreme temperature and insufficient light reduce not only flower visitation by pollinators but also pollen viability, resulting in inefficient pollination. In this study, we investigated the effects of forchlorfenuron [(N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU)] on the fruit setting and growth of oriental melons (C. melo L. var. makuwa Makino). The primary objective was to devise a new strategy for the management of oriental melons. Treatment with 5 mg·L−1 CPPU a day before flowering (T1), on the day of flowering (T2), and a day after flowering (T3) increased the fruit setting rate (by 20.1% to 30%) in melons subjected to artificial pollination (AP) or no pollination (NP) compared with the rate in those subjected to only artificial pollination without CPPU (CK). CPPU treatment induced unfertilized seeds; in addition, a tendency toward parthenocarpy was noted. The highest fruit setting rate (∼95%) was noted in plants subjected to the following treatments: AP+T1 and NP+T3. The rates of abnormal fruit formation decreased from 45.2% of CK group to 9.4% in plants subjected to AP+T1 and to 19.4% in those subjected to NP+T3. Elevated exogenous concentrations of CPPU markedly increased fruit weight. Plants subjected to NP+10 mg·L−1 CPPU bore the heaviest fruits (541.0 g), which were heavier than those borne by plants subjected to AP+10 mg·L−1 CPPU. CPPU treatment reduced the fruit cavity ratio in a concentration-dependent manner from 47.3% to 33.6% and increased the pulp thickness from 1.5 to 2.5 cm. Notably, supplementary CPPU treatment exerted no significant effects on fruit traits. Regarding taste, inconsistent results were obtained for sugar accumulation. Although the content of cucurbitacin B increased immediately after the initial CPPU treatment, it markedly decreased after 15 days of CPPU treatment. Therefore, mature melons did not have a bitter taste.

First report of Powdery Mildew Caused by Podosphaera xanthii on Siraitia grosvenorii in Guizhou, China.

Swingle (Siraitia grosvenorii), a member of the Cucurbitaceae family, stands out as a distinctive plant with both economic and medicinal significance. In October 2023, severe powdery mildew were observed on S. grosvenorii in Guiyang City (26.50°N; 106.66°E), Guizhou Province, China. About 80 % of the plants in the greenhouse showed powdery mildew symptoms. Three infected plant samples were selected for morphological and molecular analysis (GZAAS 23-0801, GZAAS 23-0802 and GZAAS 23-0803). The voucher specimens are deposited in the Key Laboratory of Agricultural Biotechnology of Guizhou Province. The symptoms initially manifested as irregular to nearly circular, small yellow spots, with distinct depressions as well as surfaces covered in white mycelium. Over time, these spots gradually expanded and merged patches. In the final stages, the entire leaves turned into yellow and withered. Microscopic observations showed that fungal hyphae were septate, branched, and flexuous to straight and 5 to 9 µm wide, and appressoria were indistinct to slightly nipple-shaped. Conidia were hyaline and ellipsoid to oval with fibrosin bodies and measured 31 to 43 × 18 to 24 μm (n = 50) with a length/width ratio of 1.3 to 2.3. Conidiophores were unbranched, straight, 120 to 268 × 14 to 22 µm (n = 30), producing two to five immature conidia in chains. Foot cells of conidiophores were cylindrical, 39 to 84 × 8 to 14 μm (n = 30), followed by one to three short cells. Short cells were cylindrical, 12 to 32 × 8 to 15 μm (n = 50). The morphological characteristics were identical with the previous description of Podosphaera xanthii (Braun and Cook, 2012). Total DNA was extracted from conidia and mycelia by the Chelex method (Walsh et al., 1991). The ribosomal DNA internal transcribed spacer (ITS) and nuclear ribosomal large subunit (LSU) were amplified by using the primers ITS1/ITS4 (White et al., 1990) and LSU1/LSU2 (Scholin et al., 1994), respectively. The ITS (OR825802, OR825803 and OR825804, respectively) and LSU (OR825805, OR825806 and OR825807, respectively) sequences of three isolates, were deposited in GenBank. The BLAST results revealed that both the ITS and LSU region sequence were 100% identical to those of P. xanthii (ITS: MF043939, MG754404 and KJ698669; LSU: OQ061319, AB936277and OP218411). Phylogenetic analyses of ITS and LSU sequences showed that our three isolates were clustered with P. xanthii (KX842351, LC270782 and LC270779) with high statistical support (ML/MP/BI: 100%/97%/1.00). Combined with their morphological characteristics, these three isolates were identified as P. xanthii. Pathogenicity tests were performed by gently brushing conidia onto the leaves of five healthy S. grosvenorii plants. Five non-inoculated plants were used as the control. All plants were maintained in a greenhouse at 25 ± 2°C. One week after inoculation, similar symptoms were observed in the inoculated plants, whereas no symptoms occurred on the control plants. By microscopic observation, the fungus on the inoculated plants was morphologically identical to those on originally diseased plants. Powdery mildew caused by P. xanthii has been reported on Vernonia cinerea (Wu et al., 2023), Vigna unguiculata (Zhang et al., 2023), Cucumis melo (Meesam et al., 2023). To our knowledge, this is the first report of powdery mildew caused by P. xanthii on S. grosvenorii in Guizhou, China. The occurrence of powdery mildew on S. grosvenorii may pose a potential threat to its large-scale cultivation. The pathogen could become a threat to other Cucurbitaceae members in the future.

Identification and functional characterization of conserved cis-regulatory elements responsible for early fruit development in cucurbit crops.

A number of cis-regulatory elements (CREs) conserved during evolution have been found to be responsible for phenotypic novelty and variation. Cucurbit crops such as cucumber (Cucumis sativus), watermelon (Citrullus lanatus), melon (Cucumis melo) and squash (Cucurbita maxima) develop fruits from an inferior ovary and share some similar biological processes during fruit development. Whether conserved regulatory sequences play critical roles in fruit development of cucurbit crops remains to be explored. In six well-studied cucurbit species, we identified 392,438 conserved non-coding sequences (CNSs), including 82,756 that are specific to cucurbits, by comparative genomics. Genome-wide profiling of accessible chromatin regions (ACRs) and gene expression patterns mapped 20,865-43,204 ACRs and their potential target genes for two fruit tissues at two key developmental stages in six cucurbits. Integrated analysis of CNSs and ACRs revealed 4,431 syntenic orthologous CNSs, including 1,687 cucurbit-specific CNSs that overlap with ACRs that are present in all six cucurbit crops and that may regulate the expression of 757 adjacent orthologous genes. CRISPR mutations targeting two CNSs present in the 1,687 cucurbit-specific sequences resulted in substantially altered fruit shape and gene expression patterns of adjacent NAC1 (NAM, ATAF1/2 and CUC2) and EXT-like (EXTENSIN-like) genes, validating the regulatory roles of these CNSs in fruit development. These results not only provide a number of target CREs for cucurbit crop improvement, but also provide insight into the roles of CREs in plant biology and during evolution.

First Report of Leaf Spot on Cucumis melo L. Caused by Arcopilus aureus in China.

Cucumis melo L. is an important fruit with widespread consumption and commercial value. However, an undescribed disease affecting Hami melon (Cucumis melo L. var. Luhoutian) plants has consistently emerged in the Qihe region of Dezhou, Shandong Province of China since 2021. The disease can occur in both seedling and mature stages of Hami melon plants, and in severely diseased areas, the incidence rate was seen as 40 to 80%. During the seedling stage, the initial symptom is the appearance of water-soaked spots on the leaves. As the disease progresses, the leaves develop necrotic spots, and severely affected plants may exhibit stem rot and decay. In the mature stage, the disease primarily affects the leaves, causing necrotic spots and chlorosis. Under conditions of high humidity, black mold can be observed in the affected areas. Small pieces of symptomatic leaves from six different infected plants were collected and surface-sterilized with 5% NaClO for 3 min and 75% alcohol for 30 s for pathogen isolation (Wang et al., 2020). After rinsing with sterile water and blotted on sterile filter paper, the tissues were established on potato dextrose agar (PDA) media and incubated at 28℃ for 3-4 days. Pure isolates showed up at PDA were obtained through single-spore isolation. Colonies of all 16 isolates obtained by single-spore isolation had similar morphological characteristics on the PDA medium, the mycelium of the isolate appears dense and yellowish-brown on the PDA medium, and also secretes a brownish-red pigment on PDA. Under the opticalmicroscope, the perithecia from PDA media are subglobose spherical in shape, 80-100 μm in diameter, brownish by reflected light, wholly and densely hairy. Terminal hairs are very dense, greyish by reflected light, olive brown to reddish brown by transmitted light, thick-walled, arcuate, circinate, or spirally coiled at the apex. The ascospores within the perithecia are elliptical or droplet-shaped, initially colorless hyaline but later becoming subhyaline slightly gray, with dimensions of 7-9 μm × 4-5 μm. The morphological characteristics of the isolates were consistent with the description of Arcopilus aureus (Wang et.al. 2016). The internal transcribed spacer (ITS) region and β-tubulin genes of three randomly selected isolates were PCR amplified and sequenced using primers ITS4/ITS5 and Bt2a/Bt2b. The sequences of ITS and β-tubulin genes were submitted to NCBI with GenBank Accession No. OR539527 and OR640972, respectively. Based on morphological features and phylogenetic analysis, we concluded that the isolates belonged to A. aureus. Pathogenicity tests were conducted by placing agar plugs-containing fungal mycelia and agar blocks (control) on leaves of Hami melon seedlings (n=12) grown at 28°C with 60% humidity in a greenhouse, the assay was repeated three times. Symptoms appeared on the pathogen-inoculated leaves seven days after inoculation, whereas the control treatment remained symptomless. The pathogens were reisolated from diseased leaves and identified as A. aureus based on morphological, and molecular phylogenetic analysis, while Koch'sostulate was used to confirm its life mode. To the best of our knowledge, this is the first report of leaf spot caused by A. aureus on Cucumis melo L. in China.

First report of cucurbit yellow stunting disorder virus infecting cucurbit crops in Jamaica.

The increasing prevalence of whitefly-transmitted viruses affecting cucurbit crops has emerged as a significant concern for global cucurbit production. Two of the most widely prevalent threats in the Americas are cucurbit yellow stunting disorder virus (CYSDV) and cucurbit chlorotic yellows virus (CCYV) (Crinivirus, Closteroviridae). These viruses induce similar foliar symptoms on cucurbit crops (Mondal et al., 2023) leading to loss of photosynthetic capability and decreased yields. Cantaloupe (Cucumis melo), watermelon (Citrullus lanatus), and cucumber (Cucumis sativus) are major cucurbit crops in St. Elizabeth, Jamaica, which is the principal fruit and vegetable producing region of the country. In August 2018, foliar symptoms were observed on cantaloupe, watermelon, and cucumber plants in several commercial farms in St. Elizabeth. These symptoms, mainly on the older leaves, consisted of severe yellowing or interveinal mottle and they appeared more pronounced on cantaloupe and cucumber plants compared to watermelon. Growers noticed the production of smaller than normal fruit. Disease incidence ranged from 10 to 100% and whiteflies (Bemisia tabaci Gennadius) were observed in the fields. To identify virus(es) associated with the disease, six plants (cantaloupe [n = 3], cucumber [n = 1] and watermelon [n = 2) exhibiting symptoms were sampled from four fields for preliminary screening. Total RNA was extracted from leaf tissues as described in Tamang et al. (2021) and samples tested by a multiplex reverse transcription RT-PCR method that targeted the RNA-dependent RNA polymerase (RdRp) of the whitefly transmitted viruses, CYSDV, CCYV, squash vein yellowing virus (SqVYV), and the aphid- transmitted cucurbit aphid-borne yellows virus (CABYV) (Mondal et al. 2023). RT-PCR amplified the expected 494-bp fragment of the CYSDV RdRp gene (Mondal et al., 2023) from two symptomatic plants; one cantaloupe, one cucumber, as well as from CYSDV-infected control plants but not from healthy controls. Further testing was conducted during the June-August 2020 growing season after similar symptoms were observed on additional farms in St. Elizabeth and two regions, Manchester and Clarendon, located to the east of St. Elizabeth. Twenty-one cucurbit leaf samples (11 cantaloupe, seven watermelon and two cucumber from St. Elizabeth and one cantaloupe from Clarendon) exhibiting foliar yellowing progressing from the crown outward, and mottling were collected. Whiteflies (5) from these fields in St. Elizabeth and 20 asymptomatic weed samples were also collected and sent to the USDA-ARS laboratory at Salinas, CA. Total RNA from leaf samples was extracted as described above and tested for CYSDV, CCYV, and CABYV. Total leaf DNA was also extracted (Mondal et al. 2016) and assayed with PCR (Gilbertson 2001) to detect the presence of the whitefly-transmitted cucurbit leaf crumple virus (CuLCrV), a begomovirus, commonly found in the southeastern United States (Gadhave et al., 2018; Keinath et al., 2018). Nineteen of the 21 cucurbit samples tested positive for the presence of CYSDV by RT-PCR (Mondal et al. 2023). Of the 19 CYSDV-positive samples, 13 cantaloupe, one cucumber, and five watermelon samples were singly infected with CYSDV, and one cantaloupe sample was infected with both CYSDV and CABYV. Amplicons of the Jamaica isolate from cantaloupe were sequenced (OR399555) and a 494 nt section of the RdRp gene was found to share 100% sequence identity to the Arizona 1 isolate (EF547827.1). The presence of CYSDV, was further confirmed using a second set of primers that amplified a 394-nt portion of the CYSDV coat protein gene (Polston et al., 2008). Among the weed samples, CABYV was detected in one sample from a Leonotis nepetifolia plant (Lamiaceae) and two Cleome sp. (Capparaceae) collected from St. Elizabeth. None of the crop and weed samples tested positive for CCYV or CuLCrV. DNA from whiteflies was extracted and assayed with PCR using species specific primers (Chen et al. 2016). All whiteflies were identified as B. tabaci cryptic species MEAM1, which is widely known an efficient vector of CYSDV (Berdiales, et al. 1999). This is the first report of CYSDV in Jamaica and its first known occurrence in these hosts within the country. Further monitoring of cucurbit crops and the whitefly vector is warranted to better understand the epidemiology.

First Report of rattail cactus necrosis-associated virus Infecting Prickly pear (Opuntia macrocentra) in the United States.

Black-spined prickly pear (Opuntia macrocentra Engelmann; Cactaceae) is a cactus native to Arizona, New Mexico, Texas, and northwest Mexico. The plant is often grown for ornamental purposes in the United States. In February 2023, virus-like symptoms such as concentric ringspots and chlorotic spots were observed on O. macrocentra plants grown at the vicinity of Maricopa County Cooperative extension, University of Arizona, Phoenix, AZ (33°24'24.6"N, 111°59'15.3"W). Total RNA was extracted from two samples (YPHC-60-A and YPHC-60-B), following the protocol by Tzanetakis et al. (2007). Reverse transcription polymerase chain reaction (RT-PCR) was performed with degenerate tobamovirus, TobamodF/TobamodR (Li et al. 2018) and potexvirus, 1RC, Potex 2RC, and Potex 5 (van der Vlugt and Berendsen 2002) primers. An expected amplicon of ~880 bp was obtained from both samples using TobamodF/TobamodR primers, while no amplification was observed with potexvirus primers. Further, RT-PCR was carried out using species-specific primers to detect cacti related tobamoviruses: cactus mild mottle virus (CMMoV), rattail cactus necrosis-associated virus (RCNaV) (Park et al. 2018) and Opuntia virus 2 (Salgado-Ortiz et al. 2020). Amplicons of ~540 bp were amplified from both samples using RCNaV specific primers, whereas no amplification was obtained using CMMoV and Opuntia virus 2 specific primers. Then, the amplicons from both YPHC-60 (A-B) isolates (~540 bp) were Sanger sequenced and shared 99.22% nucleotide identity to each other. A BLAST search revealed 93% nucleotide identity with RCNaV CP sequences (KY581586.1, JF729471, and MT130378.1). The sequences were submitted in the GenBank (accessions no. OQ914798 and OR828526). Furthermore, complete RCNaV- RNA dependent RNA polymerase (RdRP) gene was amplified using primers 3490-s-5'GTAGGTGGTACCGCATAGCA-3'; 3490as 5'AAACGCAAGTCMRYGACYGA-3' (designed in this study from accession no. JF729471.1, position 3490-3509 and 4905-4925). The expected amplicons of ~1,500 bp were obtained from both YPHC-60 (A-B) samples and sequenced (GenBank: OQ914799 and OR823954) showing 87.5 % identity with RCNaV sequences (JF729471.1 and NC_016442.1). The maximum-likelihood phylogenetic tree clustered YPHC-60 (A-B) isolates in a single clade with other RCNaV isolates. RCNaV virus particles were isolated from YPHC-60 (A-B) and submitted for RNA extraction, testing positive for RCNaV by RT-PCR. Sap extract of YPHC-60 (A-B) prepared in 0.01 M phosphate buffer (pH =7.0) was used to mechanically inoculate 3 indicator plant species (n=10): Phaseolus vulgaris, Medicago sativa, and Cucumis melo. Also, infected tissue was used to graft Opuntia sp. plants. Symptoms such as local lesions were observed on M. sativa and vein thickening on P. vulgaris 14 days post-inoculation, while Opuntia sp. showed chlorosis 30 days after grafting. RCNaV infection in mechanically inoculated P. vulgaris, M. sativa, and Opuntia sp. was also confirmed through RT-PCR. C. melo and non-inoculated control plants did not show any symptoms, nor tested positive through RT-PCR. RCNaV has been reported earlier to infect cactus species in South Korea (Park et al. 2018) and O. albicarpa in Mexico (De La Torre-Almaráz et al. 2016), where it was found in several orchards. To the best of our knowledge, this is the first report of RCNaV infecting O. macrocentra in the United States. This study highlights that RCNaV is easily transmitted mechanically or by grafting, which could impact the nursery industry as most cacti are clonally propagated.

Alteration in physio-chemical properties and gene expression pattern of snapmelon (Cucumis melo var. momordica) genotypes against drought stress

AbstractLack of water at limiting levels results in drought stress, which may have an impact on the various stages of a crop's life cycle. Four different genotypes of snapmelon (Cucumis melo L. var. momordica) responded differently to 0, 7 and 21 d of simulated drought stress. Information was collected on a range of morpho-physiological, biochemical and molecular characteristics. Each genotype had longer roots, though BAM-VR-312 had the longest roots overall. As the severity of the drought grew, the net photosynthetic rate (Pn) and stomatal conductance (Gs) dropped. In BAM-VR-312, a smaller decline in relative water content (RWC) was recorded, despite the fact that drought stress caused a significant fall in RWC. BAM-VR-312 had smaller accumulations of electrolyte leakage, hydrogen peroxide, phenol and malondialdehyde, although proline content was greater. A decrease in photosynthetic pigments was noted, though BAM-VR-312 had the least reduction. Antioxidant enzyme activity increased in BAM-VR-312, as evidenced by records of ascorbate peroxidase, catalase, guaiacol peroxidase, glutathione reductase and superoxide dismutase. Similarly, expression level of their respective genes was recorded highest in BAM-VR-312. Overall, the study clearly identified distinct genotype based on morpho-physiological, biochemical and molecular properties under drought stress and revealed that the genotype BAM-VR-312 had more efficient drought tolerance mechanisms than the other genotypes under the drought stress condition.

Species diversity, behavior and visit rates to melon flowers (Cucumis melo l.) as a function of time and seasonality in a semi-arid climate region in Brazil

In this research we evaluated the phenology of melon flowering (Cucumis melo L.) and the diversity of floral visitors in geographical conditions of semi-arid climate in Northeast Brazil, during the wet and dry seasons. The floral phenology of the melon was evaluated by recording the number of flowers produced/plant/day from 30 plants, the insects that visited the flowers were recorded following three methods: visual counts, sweep collection and counting and identification, we observe and count the number of visits to flowers from 30 plants (n = 30) during the first 45 minutes of each hour. This study reports the diversity of melon pollinators in geographical conditions located in a semi-arid climate region, with specific seasonal fluctuations, which explains the behavior pattern found at work. The order Hymenoptera predominated among floral visitors, especially Apis melífera and Trigona spinipes. The flowering cycle was influenced by seasonality, with a longer duration in the dry season, the flowering and visiting peaks were higher during the hottest hours of the day, regardless of the season. There was a preference for visitation in the morning hours, with a peak at 10 a.m. and a gradual decrease in the presence of insects in the late afternoon.

Identification and characterization analysis of the HDM gene family in melon (Cucumis melo L.).

Histone demethylase (HDM) play crucial roles in regulating plant growth and environmental adaptation. In this study, the HDM gene family in melon was identified by bioinformatics methods and the expression patterns of the CmHDM family members in different melon tissues were analyzed using transcriptome data. The results showed that 20 CmHDM genes were identified in the melon genome, which were unevenly distributed across each chromosome. These members fall into two major categories: LSD1 and JmjC. The JmjC group could be further divided into five subgroups with different numbers. The results of collinearity analysis of intraspecific and interspecific relationships showed that there were only one pair of segmental duplication in melon HDM genes, and more collinearity in genetic relationship of HDM genes between melon and tomato. The numbers of conserved domains, exons and introns in each member vary and various cis-acting elements responding to hormones and environmental signals existed in the respective promoter regions. Expression analysis showed that the respective gene members were expressed at different levels in male flowers, female flowers, roots, stems, leaves, ovary, and mature fruits of melon. These results will contribute to the understanding on the potential functions of the HDM genes and their potential functions in regulating melon growth and environmental adaptation.

Development and Application of Droplet Digital PCR Assay for the Detection of Watermelon Silver Mottle Virus and Melon Yellow Spot Virus

Watermelon silver mottle virus (WSMoV) and melon yellow spot virus (MYSV) (Tospoviridae, Orthotospovirus) are responsible for silver mottle mosaic and yellow spot symptoms, posing threats to melon (Cucumis melo), watermelon (Citrullus lanatus), and cucumber and leading to significant economic losses in China. Early disease detection and monitoring of these two viruses are necessary for disease management, for which a rapid, reliable, and adaptable diagnostic method is required. In this study, using a droplet digital PCR (ddPCR) method, the conserved region of the nucleocapsid gene (N gene) sequence was detected in WSMoV and MYSV. The probes and primers for WSMoV and MYSV did not detect other relevant cucurbit viruses, and the specificity reached 100%. Although both qPCR and ddPCR exhibited good reproducibility, the reproducibility of ddPCR was better than that of qPCR. The reproducibility of ddPCR was proved to be 100%. Moreover, ddPCR exhibited a good linear correlation with varying concentrations of targets. The detection limits of WSMoV and MYSV in ddPCR were 18 and 9 copies/μL and were approximately 12- and 18-times more than those in qPCR, respectively. Finally, 62 samples collected from the field (including infected melon, watermelon, and weeds) were further evaluated for the presence of WSMoV and MYSV. The field samples exhibited 91.94% and 51.61% positivity rates in ddPCR assays for WSMoV and MYSV, respectively; the rates were higher than those in qPCR (59.68% and 43.39%, respectively). The results indicated that ddPCR has a higher accuracy than qPCR. Therefore, ddPCR could be used in the clinical diagnosis of early infections of WSMoV and MYSV. To the best of our knowledge, this is the first study to establish a ddPCR method for the detection of WSMoV and MYSV. The application of this method for differential detection of MYSV and WSMoV will help in understanding the epidemics caused by these two important viruses and provide important information for the early detection, monitoring, and rapid extermination of infection.