Validation of candidate genes associated to earliness per se, on chromosome 5D of bread wheat

POZZI, FLORENCIA I; GHIONE, CELINA E.; HELGUERA, MARCELO; LOMBARDO, LUCIO A.; FELITTI, SILVINA A.

Rosario

Congreso; RAFV 2023 XXXIV Meeting of the Argentinean Society of Plant Physiology; 2023

Sociedad Argentina de Fisiología Vegetal

In bread wheat, earliness per se (Eps) can be defined as the residual variation in flowering time once the photoperiod and vernalization requirements have been satisfied. It has been shown that Eps genes can affect the yield of this cereal. The mapping of Eps in five environments revealed the existence of an unprecedented quantitative trait loci (QTL) QEps.imj-5D1 located on chromosome 5D, where the region most likely to find the candidate gene has a size of 26.25Mb with 391 genes annotated. In a previous work, carried out from the whole exome sequencing (MyBaits® expert panel-Arbor Biosciences) of the parent cultivars BioINTA 2001 (B01, early-heading) and Baguette Premium 11 (B11, late-heading) of contrasting phenotypes for Eps, 4 candidate genes were detected: TraesCS5D01G362700, TraesCS5D01G380900, TraesCS5D01G393200 and TraesCS5D01G401000. The objective of this work was to develop primers to validate the set of candidate genes proposed by High Resolution Melting Analysis (HRMA). The primers were developed with primer3web 4.1.0 (https://primer3.ut.ee/), the real-time PCR was performed with the CFX Maestro 1.1 software, and the data were obtained with the Precision Melt Analysis 1.3 software. Four technical replicas of each genotype were used (260 ng/μl of DNA). The markers developed allowed the differentiation between the parental B01 and B11, where the replicas of each parent are grouped into 2 different clusters. In addition, for the TraesCS5D01G401000 marker, progress was made in the differentiation between B01, B11 and artificial heterozygotes. Also, a mutant of B11 (MutP), characterized phenotypically as early-heading, grouped with B01. That is, 3 clusters were obtained: 1- replicas of B01 plus those of MutP, 2- replicas of F1Ar and 3- replicas of B11. The results obtained allow us to continue advancing in the validation of the set of proposed candidate genes by incorporating into the analysis a population of NILs previously developed by the research group.