For the primary time, the one primer enrichment know-how (SPET)—a novel high-throughput genotyping methodology—was utilized in lettuce to review the genetic variety of a set of 160 Lactuca accessions originating from 10 nations in Europe, America and Asia, and to establish genomic areas that underpin essential agronomic traits.

In latest years, applied sciences to analyze the genomic variety of crops have made extraordinary progress and novel methodologies corresponding to single primer enrichment know-how (SPET) supply promising and cost-effective alternatives. SPET has to date been utilized in a number of crop plants, corresponding to in maize, poplar, oil palm, tomato, eggplant and peach, displaying its energy for genotyping germplasm collections and crossing populations.

SPET was utilized in lettuce (Lactuca sativa L.) for the primary time by a consortium of European researchers within the context of the European Evaluation Network (EVA) of the European Cooperative Programme for Plant Genetic Resources (ECPGR), with the intention to review its genetic variety and establish genomic regions that underpin essential agronomic traits.

Lettuce is a commercially essential crop, extensively appreciated by customers for its fiber content material and low energy. It can also be a superb supply of vitamin C, iron, folate and totally different health-beneficial vitamins.

“Given the lack of cost-effective options for genotyping in lettuce, the EVA Network decided to design a SPET panel for this crop, together with IGATech, and applied it to a collection of 155 accessions of Lactuca sativa and 5 of the closely related wild species Lactuca serriola,” stated Pasquale Tripodi, lead writer of the research and Senior Researcher at CREA, Italy.

The EVA initiative was established in 2019 by ECPGR to enhance information of crop genetic variety and exploit it to breed extra resilient crops that may meet the key issues going through agriculture.

“The EVA Lettuce network is one of currently five crop-specific public–private partnerships, bringing together breeding companies, genebanks and research institutes to jointly generate phenotypic and genotypic evaluation data for numerous accessions and landraces available in European genebanks,” defined Sandra Goritschnig, coordinator of the EVA initiative and co-author of the research.

The plant supplies within the research have been chosen within the body of the EVA Lettuce Network and originated from the germplasm collections of 4 European genebanks: the Institute for Plant Genetic Resources “K.Malkov” (Bulgaria), the Centre for Genetic Resources, the Netherlands, the Unité de Génétique et Amélioration des Fruits et Légumes, Plant Biology and Breeding, INRAe (France), and the Nordic Genetic Resource Center (NordGen) (Sweden).

The studied genotypes encompassed cultivars, breeding supplies, and landraces originating from ten totally different nations in Europe, America and Asia, included totally different horticultural sorts, corresponding to Butterhead, Iceberg, Romaine, Batavia, Crisp, and have been phenotyped in multilocation subject trials throughout three nations.

Why SPET?

Compared to different genotyping strategies, SPET combines the benefits of arrays and high-throughput sequencing, and has a excessive capability to detect new single-nucleotide polymorphisms (SNPs), that are variations within the genetic sequence that decide the range amongst people in a species.

“Having a large number of SNPs increases our ability to understand the genetic diversity of a collection and investigate the function that some genomic regions play. In our case, we designed a SPET assay for 40,000 SNPs and succeeded in covering up to 96% of gene-rich regions, compared to previous studies in lettuce using different genotyping techniques, which covered only up to 27.6%. This demonstrates how effective SPET is,” stated Tripodi.

Also, utilizing a hard and fast probes panel ensures the reproducibility of the assay throughout totally different laboratories, in contrast to different sequencing strategies. It additionally fosters the institution of bigger scientific communities that may leverage interoperable markers.

The evaluation generated over 80,000 high-quality SNPs that allowed the researchers to group the accessions by sort and geographic origin, and to establish gene associations for seed coloration, leaf coloration, leaf anthocyanin content material and bolting time.

“We are very excited about this new application of SPET in lettuce, which confirmed previous findings, refined our knowledge of the genomic position of some agronomic traits and demonstrated the power of SPET to investigate the genetic diversity of germplasm collections, thus allowing a better characterization of lettuce collections. The SPET panel will be a cost-effective tool for both breeders and researchers in lettuce,” concluded Goritschnig.

The findings are printed within the journal Frontiers in Plant Science.