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Development of a kompetitive allele‐specific PCR marker for selection of the mutated Wx‐D1d allele in wheat breeding
Author(s) -
Yi Xin,
Jiang Zhengning,
Hu Wenjing,
Zhao Yun,
Bie Tongde,
Gao Derong,
Liu Datong,
Wu Ronglin,
Cheng Xiaoming,
Cheng Shunhe,
Zhang Yong
Publication year - 2017
Publication title -
plant breeding
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.583
H-Index - 71
eISSN - 1439-0523
pISSN - 0179-9541
DOI - 10.1111/pbr.12504
Subject(s) - biology , genetics , allele , locus (genetics) , amylose , glutenin , point mutation , variants of pcr , genotype , microbiology and biotechnology , mutant , gene , protein subunit , starch , biochemistry
Abstract Waxy (Wx) protein is a key enzyme for synthesis of amylose in endosperm. Amylose content in wheat grain influences the quality of end‐use products. Seven alleles have been described at the Wx‐D1 locus, but only two of them ( Wx‐D1b, Wx‐D1e ) were genotyped with codominant markers. The waxy wheat line K107Wx1 developed by treating ‘Kanto 107’ seeds with ethyl methanesulphonate carries the Wx‐D1d allele. However, no molecular basis supports this nomenclature. In the present study, DNA sequence analysis confirmed that a single nucleotide polymorphism in the sixth exon of Wx‐D1 changed tryptophan at position 301 into a termination codon. Based on this sequence variation, a PCR ‐based KASP marker was developed to detect this point mutation using 68 BC 8 F 1 plants and 297 BC 8 F 2 lines derived from the cross ‘Ningmai 14’*9/K107Wx1. Combined with codominant markers for the Wx‐A1 and Wx‐B1 alleles, waxy and non‐waxy near‐isogenic lines were distinguished. The KASP marker was efficient in identifying the mutant allele and can be used to transfer waxiness to elite lines.