SvevoZavitan_introgression_line_genotyping

Published: 16-03-2021| Version 1 | DOI: 10.17632/4vb99xs659.1
Contributor:
Harel Bacher

Description

The goal of this study was to investigate if the introduction of small wild emmer introgressions can expand the range of phenotypic response to water stress in an elite wheat cultivar background. To accomplish this, we developed a set of introgression lines (ILs) using elite durum wheat cultivar, Svevo as the backcross parent, and Zavitan as the source of wild emmer introgressions (Avni et al., 2014). Zavitan is well-adapted to the semi-arid environment and has a sequenced and annotated genome thus making it more accessible for downstream genetic analysis compared to other wild emmer accessions (Avni et al., 2017). The subset of ILs selected for this study by eliminating the wild alleles for the Reduced height (Rht)-B1b gene and non-brittle spike (TtBtr1) genes. This set included 47 wild emmer ILs were genotyped using the 90K SNP chip, and poses 1.3-14.2% of the Zavitan genes per IL.

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Steps to reproduce

The ILs were created by crossing a donor parent wild emmer (acc. Zavitan) to a recurrent parent (cv. Svevo). Homozygous RILs were obtained by selfing each plant by single seed descent method. The RILs (Avni et al., 2014) were than backcrossed to the recurrent parent for three times (BC3). As consequence contained the recurrent parent genetic background and introgressions from the Wild donor. The percentage of the recurrent parent recovery after three backcrosses was about 93.8% after filtering non informative markers (Oren, 2020). Introgression lines and parents DNA was extracted from fresh leaf tissue (*100 mg) . following a standard CTAB protocol. DNA was diluted to *50 ng/ll and was used for the Illumina genotyping array. Genotyping procedure for the 90K Chip was per- formed as described by Wang et al. (2014).