DArTseq-Based High-Throughput SilicoDArT and SNP Markers Applied for Association Mapping of Genes Related to Maize Morphology

Abstract

Today, agricultural productivity is essential to meet the needs of a growing population, and is also a key tool in coping with climate change. Innovative plant breeding technologies such as molecular markers, phenotyping, genotyping, the CRISPR/Cas method and next-generation sequencing can help agriculture meet the challenges of the 21st century more effectively. Therefore, the aim of the research was to identify single-nucleotide polymorphisms (SNPs) and SilicoDArT markers related to select morphological features determining the yield in maize. The plant material consisted of ninety-four inbred lines of maize of various origins. These lines were phenotyped under field conditions. A total of 14 morphological features was analyzed. The DArTseq method was chosen for genotyping because this technique reduces the complexity of the genome by restriction enzyme digestion. Subsequently, short fragment sequencing was used. The choice of a combination of restrictases allowed the isolation of highly informative low copy fragments of the genome. Thanks to this method, 90% of the obtained DArTseq markers are complementary to the unique sequences of the genome. All the observed features were normally distributed. Analysis of variance indicated that the main effect of lines was statistically significant (p < 0.001) for all 14 traits of study. Thanks to the DArTseq analysis with the use of next-generation sequencing (NGS) in the studied plant material, it was possible to identify 49,911 polymorphisms, of which 33,452 are SilicoDArT markers and the remaining 16,459 are SNP markers. Among those mentioned, two markers associated with four analyzed traits deserved special attention: SNP (4578734) and SilicoDArT (4778900). SNP marker 4578734 was associated with the following features: anthocyanin coloration of cob glumes, number of days from sowing to anthesis, number of days from sowing to silk emergence and anthocyanin coloration of internodes. SilicoDArT marker 4778900 was associated with the following features: number of days from sowing to anthesis, number of days from sowing to silk emergence, tassel: angle between the axis and lateral branches and plant height. Sequences with a length of 71 bp were used for physical mapping. The BLAST and EnsemblPlants databases were searched against the maize genome to identify the positions of both markers. Marker 4578734 was localized on chromosome 7, the closest gene was Zm00001d022467, approximately 55 Kb apart, encoding anthocyanidin 3-O-glucosyltransferase. Marker 4778900 was located on chromosome 7, at a distance of 45 Kb from the gene Zm00001d045261 encoding starch synthase I. The latter observation indicated that these flanking SilicoDArT and SNP markers were not in a state of linkage disequilibrium.

Unraveling the Genetic Basis of Key Agronomic Traits of Wrinkled Vining Pea (Pisum sativum L.) for Sustainable Production

“…This approach can generate SNP markers with higher numbers covering the plants’ whole genome ( Akbari et al, 2006 ; Raman et al, 2014 ; Barilli et al, 2020 ) and is successfully used in several crops, including pea ( Aznar-Fernández et al, 2020 ). The DArTseq method reduces the complexity of the genome through digestion with restriction enzymes followed by sequencing of short reads, predominantly corresponding to the active genes ( Tomkowiak et al, 2021 ). The GBS and DArTseq genotyping platforms produce high-throughput and abundant SNP markers that have been widely utilized to estimate genetic variation, discover causative allelic variations, and understand the contributing genetic architecture of complex traits with economic importance ( Baloch et al, 2017 ).…”

Genetic resources and breeding of maize for Striga resistance: a review

“…Maize populations have been genotyped using several methods, such as Random Amplified Polymorphic DNA (RAPD) ( Carvalho et al., 2004 ), RFLP ( Garcia et al., 2004 ), and AFLP ( Schrag et al., 2006 ). The SNPs are the most widely used markers for assessing the genetic diversity and association mapping in maize ( Dos Santos et al., 2016 ; Kasoma et al., 2020 ; Tomkowiak et al., 2021 ). The SNPs can be observed through various experimental protocols.…”

The Use of DArTseq Technology to Identify Markers Related to the Heterosis Effects in Selected Traits in Maize

“…5871—SNP) linked to fusarium resistance in maize have been localized within genes [ 59 ] on chromosomes 2 and 3, respectively. In their study, Tomkowiak et al [ 60 ], using DArTseq technology and association mapping, identified six markers associated with vigor and germination in maize. The existing literature indicates that four of these genes (phosphoinositide phosphatase sac7 isoform ×1 gene, grx_c8-glutaredoxin subgroup iii gene, sucrose synthase 4 isoform ×2 gene and putative SET domain-containing protein family isoform ×1 gene) determine the level of seed germination and seed vigor in maize.…”