The Gy14 cucumber (Cucumis sativus) is resistant to oomyceteous downy mildew (DM), bacterial angular leaf spot (ALS) and fungal anthracnose (AR) pathogens, but the underlying molecular mechanisms are unknown. Quantitative trait locus (QTL) mapping for the disease resistances in Gy14 and further map-based cloning identified a candidate gene for the resistant loci, which was validated and functionally characterized by spatial-temporal gene expression profiling, allelic diversity and phylogenetic analysis, as well as local association studies. We showed that the triple-disease resistances in Gy14 were controlled by the cucumber STAYGREEN (CsSGR) gene. A single nucleotide polymorphism (SNP) in the coding region resulted in a nonsynonymous amino acid substitution in the CsSGR protein, and thus disease resistance. Genes in the chlorophyll degradation pathway showed differential expression between resistant and susceptible lines in response to pathogen inoculation. The causal SNP was significantly associated with disease resistances in natural and breeding populations. The resistance allele has undergone selection in cucumber breeding. The durable, broad-spectrum disease resistance is caused by a loss-of-susceptibility mutation of CsSGR. Probably, this is achieved through the inhibition of reactive oxygen species over-accumulation and phytotoxic catabolite over-buildup in the chlorophyll degradation pathway. The CsSGR-mediated host resistance represents a novel function of this highly conserved gene in plants.

Host resistance in WI7120 cucumber to prevailing downy mildew pathogen field populations is conferred by two major-effect, one moderate-effect and two minor-effect QTL. Downy mildew (DM) caused by the obligate oomycete Pseudoperonospora cubensis is the most devastating fungal disease of cucumber worldwide. The molecular mechanism of DM resistance in cucumber is poorly understood, and use of marker-assisted breeding for DM resistance is not widely available. Here, we reported QTL mapping results for DM resistance with 243 F2:3 families from the cross between DM-resistant inbred line WI7120 (PI 330628) and susceptible '9930'. A linkage map was developed with 348 SSR and SNP markers. Phenotyping of DM inoculation responses were conducted in four field trails in 2 years at three locations. Four QTL, dm2.1, dm4.1, dm5.1, and dm6.1 were consistently and reliably detected across at least three of the four environments which together could explain 62-76 % phenotypic variations (R (2)). Among them, dm4.1 and dm5.1 were major-effect QTL (R (2) = 15-30 %) with only additive effects; dm2.1 (R (2) = 5-15 %) and dm6.1 (R (2) = 4-8 %) had moderate and minor effects, respectively. Epistatic effects were detected for dm2.1 and dm6.1 with both dm4.1 and dm5.1. One additional minor-effect QTL, dm6.2 (R (2) = 3-5 %) was only detectable with the chlorosis rating criterion. All alleles contributing to DM resistance were from WI7120. This study revealed two novel QTL for DM resistance and the unique genetic architecture of DM resistance in WI7120 conferring high level resistance to prevailing DM populations in multiple countries. The effects of disease rating scales, rating time and criteria, population size in phenotyping DM resistance on the power of QTL detection, and the use of DM resistance in WI7120 in cucumber breeding were discussed.

Map-based cloning identified a candidate gene for resistance to the anthracnose fungal pathogen Colletotrichum orbiculare in cucumber, which reveals a novel function for the highly conserved STAYGREEN family genes for host disease resistance in plants. Colletotrichum orbiculare is a hemibiotrophic fungal pathogen that causes anthracnose disease in cucumber and other cucurbit crops. No host resistance genes against the anthracnose pathogens have been cloned in crop plants. Here, we reported fine mapping and cloning of a resistance gene to the race 1 anthracnose pathogen in cucumber inbred lines Gy14 and WI 2757. Phenotypic and QTL analysis in multiple populations revealed that a single recessive gene, cla, was underlying anthracnose resistance in both lines, but WI2757 carried an additional minor-effect QTL. Fine mapping using 150 Gy14 × 9930 recombinant inbred lines and 1043 F individuals delimited the cla locus into a 32 kb region in cucumber Chromosome 5 with three predicted genes. Multiple lines of evidence suggested that the cucumber STAYGREEN (CsSGR) gene is a candidate for the anthracnose resistance locus. A single nucleotide mutation in the third exon of CsSGR resulted in the substitution of Glutamine in 9930 to Arginine in Gy14 in CsSGR protein which seems responsible for the differential anthracnose inoculation responses between Gy14 and 9930. Quantitative real-time PCR analysis indicated that CsSGR was significantly upregulated upon anthracnose pathogen inoculation in the susceptible 9930, while its expression was much lower in the resistant Gy14. Investigation of allelic diversities in natural cucumber populations revealed that the resistance allele in almost all improved cultivars or breeding lines of the U.S. origin was derived from PI 197087. This work reveals an unknown function for the highly conserved STAYGREEN (SGR) family genes for host disease resistance in plants.

The physiological responses associated with chilling of horticulturally mature cucumber (Cucumis sativus L.) fruit were examined using 13 lines that differ in chilling sensitivity. The low correlation coefficient between pitting and decay suggested that these two early manifestations of chilling injury are not significantly related. Likewise, fruit pitting and decay were not highly correlated with the tolerance of seedlings to chilling, suggesting that fruit and seedlings of the same line may have dissimilar sensitivity to chilling temperatures. Exudates from fruit cut in half transversely were collected on filter paper. The amount of exudate showed a significant correlation with pitting, decay, and percent ion leakage after 10 days of chilling. The fresh and dry weight of the exudates from fruit kept for 8 days at 12.5C ranged from 141-to 346-mg fresh weight and from 15 to 47-mg dry weight, respectively. Cucumber lines that were more sensitive to chilling had watery exudate, as indicated by their lower dry weight and percent solids. The conductivity of exudates from sensitive lines was higher (60 µsiemen/cm) than from chilling resistant lines (30 µsiemen/cm). Chilling-induced ethylene production was higher in sensitive than in resistant lines, and chilling caused a greater loss of ethylene forming enzyme activity in resistant lines than from chilling sensitive lines.

The inbred backcross line method was used to analyze the inheritance of fruit length and weight in cucumber (Cucumis sativus L.). Two populations were produced by crossing a small-fruited, adapted breeding line as the recurrent parent with 2 unadapted and large-fruited lines as donor parents. Marker genes were used in both populations to test for Mendelian expectations. Segregation was normal for all markers in both populations. Nevertheless, significant deviations from binomial expectations occurred, indicating that there were difficulties in estimating gene number and genetic variance without bias. Heritability was moderately high for fruit length and intermediate for fruit weight in both populations. No major genes controlling fruit length or weight were detected using the inbred backcross line method, and only estimates of minimum gene number were obtained. Inbred backcross lines having fruit weight equal to or greater than the large-fruited donor parent were recovered in both populations. However, in neither population were lines recovered with fruit as long as those of the donor parent.