Four soybean storage protein subunit QTLs were mapped using bulked segregant analysis and an F population, which were validated with an F RIL population. The storage protein globulins β-conglycinin (7S subunit) and glycinin (11S subunits) can affect the quantity and quality of proteins found in soybean seeds and account for more than 70% of the total soybean protein. Manipulating the storage protein subunits to enhance soymeal nutrition and for desirable tofu manufacturing characteristics are two end-use quality goals in soybean breeding programs. To aid in developing soybean cultivars with desired seed composition, an F mapping population (n = 448) and an F RIL population (n = 180) were developed by crossing high protein cultivar 'Harovinton' with the breeding line SQ97-0263_3-1a, which lacks the 7S α', 11S A, 11S A, 11S A and 11S A subunits. The storage protein composition of each individual in the F and F populations were profiled using SDS-PAGE. Based on the presence/absence of the subunits, genomic DNA bulks were formed among the F plants to identify genomic regions controlling the 7S α' and 11S protein subunits. By utilizing polymorphic SNPs between the bulks characterized with Illumina SoySNP50K iSelect BeadChips at targeted genomic regions, KASP assays were designed and used to map QTLs causing the loss of the subunits. Soybean storage protein QTLs were identified on Chromosome 3 (11S A), Chromosome 10 (7S α' and 11S A), and Chromosome 13 (11S A), which were also validated in the F RIL population. The results of this research could allow for the deployment of marker-assisted selection for desired storage protein subunits by screening breeding populations using the SNPs linked with the subunits of interest.
Identification and validation of suitable reference genes that exhibit robust transcriptional stability across many sample types is an absolute requirement of all qRT-PCR experiments. Often, however, only small numbers of reference genes, validated across limited sample types, are available for non-model species. This points to a clear need to assess and validate a wider range of potential reference genes than is currently available. We therefore looked to test and validate a large number of potential reference genes across a wide range of tissue types and treatments to determine the applicability of these reference genes for use in grapevine and other non-model plant species. Potential reference genes were selected based on stability of gene transcription in the model plant species Arabidopsis or due to their common use in the grapevine community. The selected reference genes were analyzed across two datasets consisting of a range of either 'Sauvignon blanc' or 'Pinot noir' tissues. A total of 11 potential reference genes were screened across the two datasets. Gene stability was analyzed by GeNorm, a widely used Excel application, or an ANOVA-based method developed in red clover. Both analysis methods showed that all 11 potential reference genes are stably expressed in the datasets tested, but the rankings of gene stability differed based on the datasets and analysis method used. Furthermore, the transcript stability of these genes, initially identified in Arabidopsis and now validated in grapevine, suggests applicability across a wide range of non-model plant species in addition to their utility in grapevine.
White clover (Trifolium repens L.) is a highly outcrossing heterozygous allotetraploid species, for which classic inheritance studies have been inconclusive. With the aid of molecular markers, it is now possible to study the genes controlling morphological traits. The objectives of this study were to catalog the leaf marks in white clover and map the location of leaf morphological traits based on cosegregation with molecular markers. A mapping population segregating for eight morphological traits consisting of leaf marks and number of leaflets was developed and phenotyped at two different locations during the summer and winter seasons. A confirmation population, derived by selfing one of the mapping population parents, was produced and phenotyped in one location at two different times of the year. Through the use of previously published simple sequence repeat (SSR) marker maps, linkages between the mapped molecular markers and genes for three different morphological traits was identified. The red midrib and red fleck traits were found to be controlled by two closely linked dominant genes on linkage group (LG) B1. The trifoliolate trait is controlled by at least one gene on LG H1. The identification of molecular markers linked to loci affecting leaf morphology traits resolves conflicting hypotheses on the genetics of these complex traits and has potential for molecular breeding improvement of white clover.
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