Dataset Information

A re-sequencing-based ultra-dense genetic map reveals a gummy stem blight resistance-associated gene in Cucumis melo.

ABSTRACT: The melon (Cucumis melo) genome and genetic maps with hundreds to thousands of single nucleotide polymorphism markers were recently released. However, a high-resolution genetic map was lacking. Gummy stem blight (Gsb) is a destructive disease responsible for considerable economic losses during melon production. We herein describe the development of an ultra-dense genetic map consisting of 12,932 recombination bin markers covering 1,818?cM, with an average distance of 0.17?cM between adjacent tags. A comparison of the genetic maps for melon, watermelon, and cucumber revealed chromosome-level syntenic relationships and recombination events among the three Cucurbitaceae species. Our genetic map was useful for re-anchoring the genome scaffolds of melon. More than 92% assembly was anchored to 12 pseudo-chromosomes and 90% of them were oriented. Furthermore, 1,135 recombination hotspots revealed an unbalanced recombination rate across the melon genome. Genetic analyses of the Gsb-resistant and -susceptible lines indicated the resistance phenotype is mediated by a single dominant gene. We identified Gsb-resistance gene candidates in a 108-kb region on pseudo-chromosome 4. Our findings verify the utility of an ultra-dense genetic map for mapping a gene of interest, and for identifying new disease resistant genes.

SUBMITTER: Hu Z

PROVIDER: S-EPMC5824858 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature

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A re-sequencing-based ultra-dense genetic map reveals a gummy stem blight resistance-associated gene in Cucumis melo.

Hu Zhongyuan Z Deng Guancong G Mou Haipeng H Xu Yuhui Y Chen Li L Yang Jinghua J Zhang Mingfang M

DNA research : an international journal for rapid publication of reports on genes and genomes 20180201 1

The melon (Cucumis melo) genome and genetic maps with hundreds to thousands of single nucleotide polymorphism markers were recently released. However, a high-resolution genetic map was lacking. Gummy stem blight (Gsb) is a destructive disease responsible for considerable economic losses during melon production. We herein describe the development of an ultra-dense genetic map consisting of 12,932 recombination bin markers covering 1,818 cM, with an average distance of 0.17 cM between adjacent tag ...[more]

PMID: 28985339

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Project description:Gummy stem blight caused by Stagonosporopsis cucurbitacearum is the most destructive disease of muskmelon cultivation. This study aimed to induce disease resistance against gummy stem blight in muskmelon by Trichoderma asperelloides PSU-P1. This study was arranged into two crops. Spore suspension at a concentration of 1 × 106 spores/mL of T. asperelloides PSU-P1 was applied to muskmelon to investigate gene expression. The expression of PR genes including chitinase (chi) and β-1,3-glucanase (glu) were determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and enzyme activity was assayed by the DNS method. The effects of T. asperelloides PSU-P1 on growth, yield, and postharvest quality of muskmelon fruit were measured. A spore suspension at a concentration of 1 × 106 spore/mL of T. asperelloides PSU-P1 and S. cucurbitacearum was applied to muskmelons to determine the reduction in disease severity. The results showed that the expression of chi and glu genes in T. asperelloides PSU-P1-treated muskmelon plants was 7-10-fold higher than that of the control. The enzyme activities of chitinase and β-1,3-glucanase were 0.15-0.284 and 0.343-0.681 U/mL, respectively, which were higher than those of the control (pathogen alone). Scanning electron microscopy revealed crude metabolites extracted from T. asperelloides PSU-P1-treated muskmelon plants caused wilting and lysis of S. cucurbitacearum hyphae, confirming the activity of cell-wall-degrading enzymes (CWDEs). Application of T. asperelloides PSU-P1 increased fruit weight and fruit width; sweetness and fruit texture were not significantly different among treated muskmelons. Application of T. asperelloides PSU-P1 reduced the disease severity scale of gummy stem blight to 1.10 in both crops, which was significantly lower than that of the control (2.90 and 3.40, respectively). These results revealed that application of T. asperelloides PSU-P1 reduced disease severity against gummy stem blight by overexpressed PR genes and elevated enzyme activity in muskmelon plants.

Project description:BackgroundMelon (Cucumis melo L.) is a highly diverse species that is cultivated worldwide. Recent advances in massively parallel sequencing have begun to allow the study of nucleotide diversity in this species. The Sanger method combined with medium-throughput 454 technology were used in a previous study to analyze the genetic diversity of germplasm representing 3 botanical varieties, yielding a collection of about 40,000 SNPs distributed in 14,000 unigenes. However, the usefulness of this resource is limited as the sequenced genotypes do not represent the whole diversity of the species, which is divided into two subspecies with many botanical varieties variable in plant, flowering, and fruit traits, as well as in stress response. As a first step to extensively document levels and patterns of nucleotide variability across the species, we used the high-throughput SOLiD™ system to resequence the transcriptomes of a set of 67 genotypes that had previously been selected from a core collection representing the extant variation of the entire species.ResultsThe deep transcriptome resequencing of all of the genotypes, grouped into 8 pools (wild African agrestis, Asian agrestis and acidulus, exotic Far Eastern conomon, Indian momordica and Asian dudaim and flexuosus, commercial cantalupensis, subsp. melo Asian and European landraces, Spanish inodorus landraces, and Piel de Sapo breeding lines) yielded about 300 M reads. Short reads were mapped to the recently generated draft genome assembly of the DHL line Piel de Sapo (inodorus) x Songwhan Charmi (conomon) and to a new version of melon transcriptome. Regions with at least 6X coverage were used in SNV calling, generating a melon collection with 303,883 variants. These SNVs were dispersed across the entire C. melo genome, and distributed in 15,064 annotated genes. The number and variability of in silico SNVs differed considerably between pools. Our finding of higher genomic diversity in wild and exotic agrestis melons from India and Africa as compared to commercial cultivars, cultigens and landraces from Eastern Europe, Western Asia and the Mediterranean basin is consistent with the evolutionary history proposed for the species. Group-specific SNVs that will be useful in introgression programs were also detected. In a sample of 143 selected putative SNPs, we verified 93% of the polymorphisms in a panel of 78 genotypes.ConclusionsThis study provides the first comprehensive resequencing data for wild, exotic, and cultivated (landraces and commercial) melon transcriptomes, yielding the largest melon SNP collection available to date and representing a notable sample of the species diversity. This data provides a valuable resource for creating a catalog of allelic variants of melon genes and it will aid in future in-depth studies of population genetics, marker-assisted breeding, and gene identification aimed at developing improved varieties.

Project description:Key messageWe identified QTLs associated with gummy stem blight resistance in an interspecific F2:3 Citrullus population and developed marker assays for selection of the loci in watermelon. Gummy stem blight (GSB), caused by three Stagonosporopsis spp., is a devastating fungal disease of watermelon (Citrullus lanatus) and other cucurbits that can lead to severe yield losses. Currently, no commercial cultivars with genetic resistance to GSB in the field have been reported. Utilizing GSB-resistant cultivars would reduce yield losses, decrease the high cost of disease control, and diminish hazards resulting from frequent fungicide application. The objective of this study was to identify quantitative trait loci (QTLs) associated with GSB resistance in an F2:3 interspecific Citrullus mapping population (N = 178), derived from a cross between Crimson Sweet (C. lanatus) and GSB-resistant PI 482276 (C. amarus). The population was phenotyped by inoculating seedlings with Stagonosporopsis citrulli 12178A in the greenhouse in two separate experiments, each with three replications. We identified three QTLs (ClGSB3.1, ClGSB5.1 and ClGSB7.1) associated with GSB resistance, explaining between 6.4 and 21.1% of the phenotypic variation. The genes underlying ClGSB5.1 includes an NBS-LRR gene (ClCG05G019540) previously identified as a candidate gene for GSB resistance in watermelon. Locus ClGSB7.1 accounted for the highest phenotypic variation and harbors twenty-two candidate genes associated with disease resistance. Among them is ClCG07G013230, encoding an Avr9/Cf-9 rapidly elicited disease resistance protein, which contains a non-synonymous point mutation in the DUF761 domain that was significantly associated with GSB resistance. High throughput markers were developed for selection of ClGSB5.1 and ClGSB7.1. Our findings will facilitate the use of molecular markers for efficient introgression of the resistance loci and development of GSB-resistant watermelon cultivars.

Dataset Information

A re-sequencing-based ultra-dense genetic map reveals a gummy stem blight resistance-associated gene in Cucumis melo.

Publications

A re-sequencing-based ultra-dense genetic map reveals a gummy stem blight resistance-associated gene in Cucumis melo.

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