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Combining a New Exome Capture Panel With an Effective varBScore Algorithm Accelerates BSA-Based Gene Cloning in Wheat
Ist Teil von
Frontiers in plant science, 2020-08, Vol.11, p.1249-1249
Ort / Verlag
Frontiers Media S.A
Erscheinungsjahr
2020
Quelle
EZB Free E-Journals
Beschreibungen/Notizen
The discovery of functional genes underlying agronomic traits is of great importance for wheat improvement. Here we designed a new wheat exome capture probe panel based on IWGSC RefSeq v1.0 genome sequence information and developed an effective algorithm, varBScore, that can sufficiently reduce the background noise in gene mapping and identification. An effective method, termed
b
ulked
s
egregant
e
xome capture
seq
uencing (BSE-Seq) for identifying causal mutations or candidate genes was established by combining the use of a newly designed wheat exome capture panel, sequencing of bulked segregant pools from segregating populations, and the robust algorithm varBScore. We evaluated the effectiveness of varBScore on SNP calling using the published dataset for mapping and cloning the yellow rust resistance gene
Yr7
in wheat. Furthermore, using BSE-Seq, we rapidly identified a wheat yellow leaf mutant gene,
ygl1
, in an ethyl methanesulfonate (EMS) mutant population and found that a single mutation of G to A at 921 position in the wild type
YGL1
gene encoding magnesium-chelatase subunit chlI caused the leaf yellowing phenotype. We further showed that mutation of
YGL1
through CRISPR/Cas9 gene editing led to a yellow phenotype on the leaves of transgenic wheat, indicating that
ygl1
is the correct causal gene responsible for the mutant phenotype. In summary, our approach is highly efficient for discovering causal mutations and gene cloning in wheat.