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Polyploid QTL-seq towards rapid development of tightly linked DNA markers for potato and sweetpotato breeding through whole-genome resequencing.

ABSTRACT: Potato (Solanum tuberosum L.) and sweetpotato (Ipomoea batatas L.), which are nutritionally and commercially important tuberous crops, possess a perplexing heredity because of their autopolyploid genomes. To reduce cross-breeding efforts for selecting superior cultivars from progenies with innumerable combinations of traits, DNA markers tightly linked to agronomical traits are required. To develop DNA markers, we developed a method for quantitative trait loci (QTL) mapping using whole-genome next-generation sequencing (NGS) in autopolyploid crops. To apply the NGS-based bulked segregant method, QTL-seq was modified. (1) Single parent-specific simplex (unique for one homologous chromosome) single-nucleotide polymorphisms (SNPs), which present a simple segregation ratio in the progenies, were exploited by filtering SNPs by SNP index (allele frequency). (2) Clusters of SNPs, which were inherited unevenly between bulked progenies with opposite phenotypes, especially those with an SNP index of 0 for the bulk that did not display the phenotypes of interest, were explored. These modifications allowed for separate tracking of alleles located on each of the multiple homologous chromosomes. By applying this method, clusters of SNPs linked to the potato cyst nematode resistance H1 gene and storage root anthocyanin (AN) content were identified in tetraploid potato and hexaploid sweetpotato, respectively, and completely linked DNA markers were developed at the site of the presented SNPs. Thus, polyploid QTL-seq is a versatile method that is free from specialized manipulation for sequencing and construction of elaborate linkage maps and facilitates rapid development of tightly linked DNA markers in autopolyploid crops, such as potato and sweetpotato.

SUBMITTER: Yamakawa H

PROVIDER: S-EPMC8486255 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:PurposeIn Uganda, sweetpotato [Ipomoea batatas (L.) Lam] is typically a "woman's crop," grown, processed, stored and also mainly consumed by smallholder farmers for food and income. Farmers value sweetpotato for its early maturity, resilience to stresses, and minimal input requirements. However, productivity remains low despite the effort of breeding programs to introduce new varieties. Low uptake of new varieties is partly attributed to previous focus by breeders on agronomic traits and much less on quality traits and the diverse preferences of men and women in sweetpotato value chains.MethodTo address this gap, breeders, food scientists, and social scientists (including gender specialists) systematically mainstreamed gender into the breeding program. This multidisciplinary approach, grounded in examining gender roles and their relationship with varietal and trait preferences, integrated important traits into product profiles.ResultsBuilding on earlier efforts of participatory plant breeding and participatory varietal selection, new interventions showed subtle but important gender differences in preferences. For instance, in a study for the RTBFoods project, women prioritized mealiness, sweetness, firmness and non-fibrous boiled roots. These were further subjected to a rigorous gender analysis using the G+ product profile query tool. The breeding pipelines then incorporated these gender-responsive priority quality traits, prompting the development of standard operating procedures to phenotype these traits.ConclusionFollowing an all-inclusive approach coupled with training of multidisciplinary teams involving food scientists, breeders, biochemists, gender specialists and social scientists, integration into participatory variety selection in Uganda enabled accentuation of women and men's trait preferences, contributing to clearer breeding targets. The research has positioned sweetpotato breeding to better respond to the varying needs and preferences of the users.

Project description:Given the importance of prioritizing genome-based breeding of sweet potato to enable the promotion of food and nutritional security for future human societies, here, we aimed to dissect the genetic basis of storage root starch content (SC) when associated with a complex set of breeding traits including dry matter (DM) rate, storage root fresh weight (SRFW), and anthocyanin (AN) content in a mapping population containing purple-fleshed sweet potato. A polyploid genome-wide association study (GWAS) was extensively exploited using 90,222 single-nucleotide polymorphisms (SNPs) obtained from a bi-parental 204 F1 population between 'Konaishin' (having high SC but no AN) and 'Akemurasaki' (having high AN content but moderate SC). Through the comparison of polyploid GWAS on the whole set of the 204 F1, 93 high-AN-containing F1, and 111 low-AN-containing F1 populations, a total of two (consists of six SNPs), two (14 SNPs), four (eight SNPs), and nine (214 SNPs) significantly associated signals were identified for the variations of SC, DM, SRFW, and the relative AN content, respectively. Of them, a novel signal associated with SC, which was most consistent in 2019 and 2020 in both the 204 F1 and 111 low-AN-containing F1 populations, was identified in homologous group 15. The five SNP markers associated with homologous group 15 could affect SC improvement with a degree of positive effect (~4.33) and screen high-starch-containing lines with higher efficiency (~68%). In a database search of 62 genes involved in starch metabolism, five genes including enzyme genes granule-bound starch synthase I (IbGBSSI), α-amylase 1D, α-amylase 1E, and α-amylase 3, and one transporter gene ATP/ADP-transporter were located on homologous group 15. In an extensive qRT-PCR of these genes using the storage roots harvested at 2, 3, and 4 months after field transplantation in 2022, IbGBSSI, which encodes the starch synthase isozyme that catalyzes the biosynthesis of amylose molecule, was most consistently elevated during starch accumulation in sweet potato. These results would enhance our understanding of the underlying genetic basis of a complex set of breeding traits in the starchy roots of sweet potato, and the molecular information, particularly for SC, would be a potential platform for molecular marker development for this trait.

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Polyploid QTL-seq towards rapid development of tightly linked DNA markers for potato and sweetpotato breeding through whole-genome resequencing.

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