Phytophthora capsici Leonian, the causal agent of foliar blight, root rot, fruit rot and crown rot syndromes in squash (Cucurbita moschata), is a devastating pathogen worldwide. Resistance to Phytophthora crown rot in University of Florida breeding line #394-1-27-12 (C. moschata) is conferred by three independent dominant genes (R1R2R3). Availability of DNA markers linked to R1R2R3 genes would allow efficient breeding for Phytophthora crown rot resistance through marker-assisted selection (MAS). The goal of the current study was to identify quantitative trait loci (QTLs) associated with resistance to Phytophthora crown rot in an F₂ population (n = 168) derived from a cross between #394-1-27-12 (R) and Butter Bush (S) using QTL-seq bulk segregant analysis. Whole-genome resequencing of the resistant (n = 20) and susceptible (n = 20) bulk segregants revealed ~900,000 single nucleotide polymorphisms distributed across C. moschata genome. Three QTLs significantly (P < 0.05) associated with resistance to Phytophthora crown rot were detected on chromosome 4 (QtlPC-C04), 11 (QtlPC-C11) and 14 (QtlPC-C14). Several markers linked to these QTLs are potential targets for MAS against Phytophthora crown rot in C. moschata. The present study reports the first QTLs associated with Phytophthora crown rot resistance in C. moschata.

In plants, resistance genes (R) are key players in combatting diseases caused by various phytopathogens. Typically, resistance relies on detecting a single pathogen-derived molecular pattern. However, R-gene-mediated resistance is often race specific, follows the gene-for-gene hypothesis, and can be overcome in field conditions as pathogens evolve. On the contrary, altering plant susceptibility genes (S-genes) facilitates compatibility and results in broad and durable resistance. S-genes are negative regulators present in plants and exploited by pathogens to facilitate their growth and cause infection. Several studies across crop species have reported manipulation of S-genes using genome editing to confer broad spectrum resistance. This review focuses on the plant defense mechanism against biotic stress, R-genes vs. S-genes, different types/classes of S-genes, different tools for S-gene discovery, and the use of gene editing technologies to target S-genes in addition to their applications, challenges, and future perspectives.

Zucchini Yellow Mosaic Virus (ZYMV) is an aphid-transmitted potyvirus that causes severe yield losses in squash (Cucurbita moschata) production worldwide. Development of resistant cultivars using traditional breeding approaches relies on rigorous and resource-intensive phenotypic assays. QTL-seq, a whole genome re-sequencing based bulked segregant analysis, is a powerful tool for mapping quantitative trait loci (QTL) in crop plants. In the current study, the QTL-seq approach was used to identify genetic loci associated with ZYMV resistance in an F₂ population (n = 174) derived from a cross between Nigerian Local (resistant) and Butterbush (susceptible). Whole genome re-sequencing of the parents and bulks of resistant and susceptible F₂ progeny revealed a mapping rate between 94.04% and 98.76%, and a final effective mapping depth ranging from 81.77 to 101.73 across samples. QTL-seq analysis identified four QTLs significantly (p < 0.05) associated with ZYMV resistance on chromosome 2 (QtlZYMV-C02), 4 (QtlZYMV-C04), 8 (QtlZYMV-C08) and 20 (QtlZYMV-C20). Seven markers within the QTL intervals were tested for association with ZYMV resistance in the entire F₂ population. For QtlZYMV-C08, one single nucleotide polymorphism (SNP) marker (KASP-6) was found to be significantly (p < 0.05) associated with ZYMV resistance, while two SNPs (KASP-1 and KASP-3) and an indel (Indel-2) marker were linked to resistance within QtlZYMV-C20. KASP-3 and KASP-6 are non-synonymous SNPs leading to amino acid substitutions in candidate disease resistant gene homologs on chromosomes 20 (CmoCh20G003040.1) and 8 (CmoCh08G007140.1), respectively. Identification of QTL and SNP markers associated with ZYMV resistance will facilitate marker-assisted selection for ZYMV resistance in squash.

Phytophthora crown rot, caused by Phytophthora capsici Leonian, is a devastating disease in commercial squash ( Cucurbita pepo L.) production across the United States. Current management practices rely heavily on the use of chemical fungicides, but existence of fungicide-resistant pathogen populations has rendered many chemicals ineffective. Host resistance is the best strategy for managing this disease; however, no commercial cultivars resistant to the pathogen are currently available. Resistance to Phytophthora crown rot in PI 181761 ( C. pepo ) is an important genetic resource for squash breeders worldwide; however, the underlying genetic basis of resistance in PI 186761 that would allow designing of sound breeding strategies is currently unknown. The goal of the current study was to determine the inheritance of resistance in breeding line #186761-36P, a resistant selection of PI 181761, using phenotypic data from F ₁ , F ₂ , and backcross populations derived from a cross between #181761-36P and a susceptible acorn-type cultivar, Table Queen. The results indicated that resistance in #181761-36P is controlled by three dominant genes (R4, R5, and R6). Introgression of these genes into susceptible cultivar groups of C. pepo will provide an important tool in the integrated management of Phytophthora crown rot.

Phytophthora capsici Leonian, the causal agent of Phytophthora crown rot in squash ( Cucurbita pepo L.), is an economically important pathogen worldwide. Currently, no C. pepo cultivars immune to the pathogen are commercially available, but sources of resistance to Phytophthora crown rot have been identified in a set of 16 C. pepo plant introductions (PIs). Knowledge of the genetic relationships among these accessions and their relatedness to economically important morphotypes of C. pepo would inform breeders’ best strategies for introgressing resistance; however, this information is currently lacking. The goal of the current study was to determine genetic diversity among the resistant accessions and their genetic relatedness to susceptible morphotypes of subspecies pepo (Zucchini and Pumpkin) and texana (Acorn, Straightneck, and Crookneck) using 39 SSR markers. The markers revealed 132 alleles averaging 4.40 alleles per locus and had a mean polymorphic information content (PIC) and gene diversity of 0.44 and 0.49, respectively. CMTp235 had the highest PIC and gene diversity of 0.80 and 0.82, respectively. Hierarchical clustering by UPGMA and principal coordinate analysis (PCOA) revealed grouping into two major C. pepo subspecies, texana and pepo , with all the resistant accessions grouping in the latter. In order of increasing genetic distance (GD), the resistant accessions were least distant to Zucchini (GD = 0.34), followed by Pumpkin (GD = 0.40), Crookneck (GD = 0.56), Acorn (GD = 0.60), and Straightneck (GD = 0.61) morphotypes. Mean GD among the resistant accessions was 0.31 and was highest between PIs 615142 and 615132 (0.61). Based on genetic similarity, PIs 174185 and 181761 (disease severity ≤1.4) would be the best sources of resistance for transfer into subspecies texana and pepo , respectively. Overall, the results presented here support a closer genetic relationship between the resistant accessions and morphotypes of subspecies pepo than those of subspecies texana .

Pumpkin (Cucurbita pepo) seeds are nutritious and valued as a source of vegetable oil, protein, healthy fatty acids, and minerals. Pumpkin seeds that are naturally devoid of the seedcoat (hull-less) are preferred by the industry as they eliminate the need for de-hulling prior to use. A single recessive gene, designated as n or h, controls the hull-less seed trait in pumpkin. Visual selection for the trait is easy, however, it is resource intensive when applied to large breeding populations. High throughput genotyping assays can aid in the identification of suitable individuals in segregating populations through marker-assisted selection. In the current study, the QTL-seq approach was used to identify genetic loci, SNP markers and candidate genes associated with the hull-less trait in a segregating F₂ population (n = 143) derived from a cross between Kakai (hull-less) × Table Gold Acorn (hulled). The segregation of the hull-less trait in the F₂ population fit a 3:1 ratio (p < 0.05). QTL-seq analysis detected a single QTL on chromosome 12 (Qtlhull-less-C12) which was significantly associated with the hull-less trait in C. pepo. Twenty-eight SNPs were genotyped in the population, two among which (Ch12_3412046 and Ch12_3417142) were significantly associated (p < 0.05) with the hull-less trait in cultivars and accessions of diverse genetic background. Several candidate genes fall within the Qtlhull-less-C12 interval, among them is the No Apical meristem (NAC) domain-containing protein and a Fiber Protein fb11 gene involved in lignin accumulation and cell wall deposition across plant species, respectively. The findings of this study will facilitate the marker-assisted selection for the hull-less seed trait in pumpkin and further our understanding of the functional mechanisms underlying the trait across cucurbit crops.