Unknown,Transcriptomics,Genomics,Proteomics

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Analysis of the 4AL QTL for grain dormancy in wheat


ABSTRACT: We generated multiple NILs segregating for the 4A-1 QTL following a method based on HIFs analysis. Five F7 RILs from the mapping population were chosen as founders of HIFs on the basis of being heterozygote for the 4A-1 QTL region, while maximizing homozygosity for the background genome (about 97 % homozygosis). From them, five F8 HIFs were obtained by self-pollination and were phenotyped and genotyped. In every HIF, homozygote individuals for the Yitpi allele showed low GI while homozygous for the other alleles showed high GI. Heterozygous individuals showed intermediate GI values in at least four out of the five HIFs. From each HIF, homozygous individuals with contrasting QTL effects were selected and self-pollinated to generate five F9 sets of NILs. Those NILs were finally grown together with the four parent varieties, and grain samples collected at 15, 25 and 35 days post-anthesis (DPA). RNA was isolated from those samples and pooled together in two groups, one containing all samples coming from NILs carrying the QTL (pool A) and the other containing all samples coming from NILs without the QTL (pool B). Samples collected from the four parents at the same DPA were also included in the corresponding RNA pool. This innovative pooling approach allowed us to distinguish a real signal coming from the targeted selected region (segregating equally in each set of NILs) while background noise coming from the rest of the genome is neutralized (different in every set of NILs).

INSTRUMENT(S): Illumina HiSeq 2000

ORGANISM(S): Triticum aestivum

SUBMITTER: Jose Barrero 

PROVIDER: E-MTAB-4254 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Transcriptomic analysis of wheat near-isogenic lines identifies PM19-A1 and A2 as candidates for a major dormancy QTL.

Barrero Jose M JM   Cavanagh Colin C   Verbyla Klara L KL   Tibbits Josquin F G JF   Verbyla Arunas P AP   Huang B Emma BE   Rosewarne Garry M GM   Stephen Stuart S   Wang Penghao P   Whan Alex A   Rigault Philippe P   Hayden Matthew J MJ   Gubler Frank F  

Genome biology 20150512


<h4>Background</h4>Next-generation sequencing technologies provide new opportunities to identify the genetic components responsible for trait variation. However, in species with large polyploid genomes, such as bread wheat, the ability to rapidly identify genes underlying quantitative trait loci (QTL) remains non-trivial. To overcome this, we introduce a novel pipeline that analyses, by RNA-sequencing, multiple near-isogenic lines segregating for a targeted QTL.<h4>Results</h4>We use this approa  ...[more]

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