Allele mining of TaGRF-2D gene 5’-UTR in Triticum aestivum and Aegilops tauschii genotypes

Abstract

The low diversity of the D-subgenome of bread wheat requires the involvement of new alleles for breeding. In grasses, the allelic state of Growth Regulating Factor (GRF) gene is correlated with nitrogen uptake. In this study, we characterized the sequence of TaGRF-2D and assessed its diversity in bread wheat and goatgrass Aegilops tauschii (genome DD). In silico analysis was performed for reference sequence searching, primer pairs design and sequence assembly. The gene sequence was obtained using Illumina and Sanger sequencing. The complete sequences of TaGRF-2D were obtained for 18 varieties of wheat. The polymorphism in the presence/absence of two GCAGCC repeats in 5' UTR was revealed and the GRF-2D-SSR marker was developed. Our results showed that the alleles 5' UTR-250 and 5' UTR-238 were present in wheat varieties, 5' UTR-250 was presented in the majority of wheat varieties. In Ae. tauschii ssp. strangulata (likely donor of the D-subgenome of polyploid wheat), most accessions carried the 5' UTR-250 allele, whilst most Ae. tauschii ssp. tauschii have 5' UTR-244. The developed GRF-2D-SSR marker can be used to study the genetic diversity of wheat and Ae. tauschii.

Effect of Gibberellin Responsive Reduced Height Allele Rht13 on Agronomic Traits in Spring Bread Wheat in Field Experiment in Non-Black Soil Zone

“…In our study, the presence of Rht13 reduced thousand grain weight by 9.8%, however, it increased seed number per spikelet and, as a result, seed number per spike. The introduction of the alleles of genes and quantitative trait loci associated with higher thousand grain weight could improve this drawback in the future [26,27].…”

Alleles of the GRF3-2A Gene in Wheat and Their Agronomic Value

“…The reverse primer was 5 -labelled with 6-carboxyfluorescein. As predicted by gene sequencing, the length of the PCR products for the TaGRF3-2A gene was expected in range 230 to 282 bp, whereas for TaGRF3-2D the expected sizes were 211 to 223 bp [51]. Thus, the PCR-product sizes of TaGRF3-2A and TaGRF3-2D genes do not overlap.…”

TaGRF3-2A Improves Some Agronomically Valuable Traits in Semi-Dwarf Spring Triticale

“…Additionally, we used CAPS marker for the detection of the dwarfing Ddw1 allele [52]. To detect polymorphism in the microsatellite locus of the 5'-untranslated region in Grf3, we designed primers F: 5'-GTAGGAGTAAAAGGCAAAAGCACG-3' and R: 5'-ACAGGGAGGCAAAGGGCATC-3' (Figure 6) [23,31]. The alleles of Grf3 were assayed by PCR with subsequent fragment analysis performed using a 3130xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).…”

“…In common wheat, 30 TaGRF genes were identified on 12 chromosomes, which were divided into four phylogenetic groups [21,22]. Analysis of the TaGRF3 gene sequence on the 2D common wheat chromosome using molecular markers showed its allelic diversity among the accessions of the collection, as well as a correlation with thousand grain weight and the size of wheat kernels [23]. A number of authors propose that it is the tuning of genes of nitrogen metabolism in low-stemmed varieties that will drive a new wave of the green revolution, increasing the absorption of nitrogen and, as a consequence, the weight of the grain [19,24,25].…”