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Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

Published on: March 31, 2017

Genetic mapping of powdery mildew resistance genes in soybean by high-throughput genome-wide sequencing

Bingzhi Jiang^1,2, Mu Li^1,2, Yanbo Cheng^1,2

¹The State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, South China Agricultural University, Guangzhou, 510642, Guangdong, People's Republic of China.

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik

|March 4, 2019

Abstract

Key Message:

The Mendelian locus conferring resistance to powdery mildew in soybean was precisely mapped using a combination of phenotypic screening, genetic analyses, and high-throughput genome-wide sequencing. Powdery mildew (PMD), caused by the fungus Microsphaera diffusa Cooke & Peck, leads to considerable yield losses in soybean [Glycine max (L.) Merr.] under favourable environmental conditions and can be controlled by identifying germplasm resources with resistance genes. In this study, resistance to M. diffusa among resistant varieties B3, Fudou234, and B13 is mapped as a single Mendelian locus using three mapping populations derived from crossing susceptible with resistant cultivars. The position of the PMD resistance locus in B3 is located between simple sequence repeat (SSR) markers GMES6959 and Satt_393 on chromosome 16, at genetic distances of 7.1 cM and 4.6 cM, respectively. To more finely map the PMD resistance gene, a high-density genetic map was constructed using 248 F₈ recombinant inbred lines derived from a cross of Guizao1 × B13. The final map includes 3748 bins and is 3031.9 cM in length, with an average distance of 0.81 cM between adjacent markers. This genotypic analysis resulted in the precise delineation of the B13 PMD resistance locus to a 188.06-kb genomic region on chromosome 16 that harbours 28 genes, including 17 disease resistance (R)-like genes in the reference Williams 82 genome. Quantitative real-time PCR assays of possible candidate genes revealed differences in the expression levels of 9 R-like genes between the resistant and susceptible parents. These results provide useful information for marker-assisted breeding and gene cloning for PMD resistance.

Insights

Researchers precisely mapped soybean powdery mildew resistance to a specific gene locus on chromosome 16. This finding aids in developing disease-resistant soybean varieties through marker-assisted breeding.

Area of Science:

Plant Genetics
Crop Science
Molecular Biology

Background:

Powdery mildew (PMD) causes significant soybean yield losses.
Identifying soybean germplasm with resistance genes is crucial for disease control.
Previous studies indicated a Mendelian locus confers resistance to PMD.

Purpose of the Study:

To precisely map the Mendelian locus conferring resistance to powdery mildew in soybean.
To identify candidate genes within the mapped region for PMD resistance.
To provide foundational data for marker-assisted breeding and gene cloning.

Main Methods:

Phenotypic screening and genetic analyses of three mapping populations.
High-throughput genome-wide sequencing and construction of a high-density genetic map.
Quantitative real-time PCR to analyze gene expression in resistant and susceptible soybean parents.

Main Results:

The PMD resistance locus was mapped to chromosome 16, between SSR markers GMES6959 and Satt_393.
Fine mapping delineated the locus to a 188.06-kb region containing 28 genes, including 17 disease resistance (R)-like genes.
Differential expression of 9 R-like genes was observed between resistant and susceptible soybean parents.

Conclusions:

Precise mapping of the soybean PMD resistance locus provides a valuable resource for breeding programs.
The identified genomic region and candidate R-like genes offer targets for developing improved soybean cultivars.
This research facilitates marker-assisted breeding and gene cloning for enhanced soybean powdery mildew resistance.

Read full analysis

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