Fruit and vegetable crops are rich in dietary fibre, vitamins and minerals, which are vital to human health. However, many biotic stressors (such as pests and diseases) and abiotic stressors threaten crop growth, quality, and yield. Traditional breeding strategies for improving crop traits include a series of backcrosses and selection to introduce beneficial traits into fine germplasm, this process is slow and resource-intensive. The new breeding technique known as clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein-9 (Cas9) has the potential to improve many traits rapidly and accurately, such as yield, quality, disease resistance, abiotic stress tolerance, and nutritional aspects in crops. Because of its simple operation and high mutation efficiency, this system has been applied to obtain new germplasm resources via gene-directed mutation. With the availability of whole-genome sequencing data, and information about gene function for important traits, CRISPR-Cas9 editing to precisely mutate key genes can rapidly generate new germplasm resources for the improvement of important agronomic traits. In this review, we explore this technology and its application in fruit and vegetable crops. We address the challenges, existing variants and the associated regulatory framework, and consider future applications.

Melon (Cucumis melo L.) is a protected crop in China with high economic value. Agrobacterium-mediated genetic transformation is a powerful tool to improve agronomic traits and obtain elite germplasm. However, current transformation protocols in melons are inefficient and highly genotype-dependent. To improve transformation in melon, we tested different infiltration methods for Agrobacterium-mediated transformation. Among these methods, micro-brushing and sonication for 20 s, followed by vacuum infiltration at −1.0 kPa for 90 s, resulted in the strongest green fluorescent protein signal and increased the proportion of infected explants. We transformed melon with developmental regulatory genes AtGRF5, AtPLT5, AtBBM, AtWUS, AtWOX5, and AtWIND1 from Arabidopsis and estimated regeneration frequencies as the number of regenerating shoots/total number of inoculated explants in the selection medium. The overexpression of AtGRF5 and AtPLT5 in melon resulted in transformation efficiencies of 42.3% and 33% in ZHF and 45.6% and 32.9% in Z12, respectively, which were significantly higher than those of the control. AtGRF5 and AtPLT5 expression cassettes were added to CRISPR/Cas9 genome-editing vectors to obtain transgenic phytoene desaturase CmPDS knockout mutants. Using AtGRF5 or AtPLT5, multi-allelic mutations were observed at CmPDS target sites in recalcitrant melon genotypes. This strategy enables genotype-flexible transformation and promotes precise genome modification technologies in melons.

CRISPR/Cas9 is a powerful genome editing tool for trait improvement in various crops; however, enhancing mutation efficiency using CRISPR/Cas9 in watermelon and melon remains challenging. We designed four CRISPR systems with different sgRNA expression cassettes to target the phytoene desaturase (PDS) gene in melon. The constructed vectors were delivered to host plants using Agrobacterium-mediated transformation. Phenotypic and genotypic analyses of the edited melon seedlings revealed that the CRISPR systems with tRNA and Csy4 spacers driven by the Pol II-type promoter significantly improved mutation efficiency, reaching 25.20% and 42.82%, respectively. Notably, 78.95% of the mutations generated by the Csy4 system involved large-fragment deletions (LDs) between the two target sites. In watermelon, the Csy4 system achieved a PDS editing efficiency of 41.48%, with 71.43% of the edited seedlings showing LD between the two target sites. Sequencing analysis indicated that the edited melon seedlings exhibited heterozygous, three-allele mutation and chimeric events; the edited watermelon seedlings included 2/14 homozygous mutations. Compared to the commonly used Pol III promoter, using the Pol II promoter to drive sgRNA expression cassettes containing Csy4 showed the best improvement in CRISPR/Cas9 editing efficiency in melon; this system was also effective in watermelon.

Powdery mildew (PM), a common disease of many major crop species, including melon (Cucumis melo L.), affects plant growth and fruit quality and seriously reduces production. Using a combined morphological and molecular approach, we attribute the PM pathogen that naturally occurs in melon to Podosphaera xanthii, and specifically to physiological race 1. An investigation into the genetic basis of PM resistance in melon using the resistant accession ‘PI 164637’ and susceptible counterpart ‘HDZ’ reveals dominant inheritance of PM resistance at the seedling stage, supported by F₂ and backcross population segregation ratios. Adult plant assessments indicate a major gene with an additive effect for PM resistance. Bulk segregant analysis coupled with high-throughput sequencing identified a significant quantitative trait locus on chromosome 6 that is associated with PM resistance. Genetic mapping narrowed down the candidate region to 63.5 kb using InDel molecular markers, harboring 12 candidate genes. The marker chr06_indel_5 047 127 demonstrated high accuracy in screening PM resistance in an F₂ segregating population and 30 inbred lines as natural populations. Functional annotation and expression analysis of candidate genes revealed that MYB transcription factor MELO3C006700, GATA transcription factor MELO3C028829 and heparanase-like protein MELO3C006697 are promising candidate genes for PM resistance in melon. The genetic architecture underlying this resistance in melon offers valuable insights for breeding programs, and the identified markers, especially chr06_indel_5 047 127, may enable practical applications for marker-assisted selection in developing PM-resistant melon varieties.

Earliness and plant architecture are major breeding targets in cucurbit crops because they influence crop duration, early yield, planting density, canopy management, and production efficiency. This review summarizes genetic, molecular, and genome-editing advances related to flowering, sex expression, fruit development, stem elongation, branching, and canopy architecture in cucumber, melon, watermelon, Cucurbita species, and other cucurbits. Current evidence shows that earlier flowering or increased femaleness does not necessarily lead to earlier commercial maturity, while compact growth must be evaluated together with yield, fruit quality, and environmental adaptation. CRISPR/Cas9 knockout remains the most established editing approach, whereas base editing, cis-regulatory editing, multiplex editing, developmental-regulator-assisted regeneration, and DNA-free RNP delivery offer promising routes toward more precise trait optimization. Major constraints include genotype-dependent transformation, pleiotropic effects, limited validation in elite germplasm, and scarce direct evidence for shortening fruit-development duration. An evidence-guided framework is proposed that links trait definition, functional genomics, target prioritization, tailored genome editing, whole-plant evaluation, and breeding deployment. Future progress will depend on quantitatively tuned alleles and coordinated optimization of reproductive timing, canopy architecture, productivity, fruit quality, and environmental stability.

Earliness and plant architecture are major breeding targets in cucurbit crops because they influence crop duration, early yield, planting density, canopy management, and production efficiency. This review summarizes genetic, molecular, and genome-editing advances related to flowering, sex expression, fruit development, stem elongation, branching, and canopy architecture in cucumber, melon, watermelon, Cucurbita species, and other cucurbits. Current evidence shows that earlier flowering or increased femaleness does not necessarily lead to earlier commercial maturity, while compact growth must be evaluated together with yield, fruit quality, and environmental adaptation. CRISPR/Cas9 knockout remains the most established editing approach, whereas base editing, cis-regulatory editing, multiplex editing, developmental-regulator-assisted regeneration, and DNA-free RNP delivery offer promising routes toward more precise trait optimization. Major constraints include genotype-dependent transformation, pleiotropic effects, limited validation in elite germplasm, and scarce direct evidence for shortening fruit-development duration. An evidence-guided framework is proposed that links trait definition, functional genomics, target prioritization, tailored genome editing, whole-plant evaluation, and breeding deployment. Future progress will depend on quantitatively tuned alleles and coordinated optimization of reproductive timing, canopy architecture, productivity, fruit quality, and environmental stability.