Allele-specific PCR Markers for Distinguishing High Molecular Weight Glutenin Subunit Dtx5 of Aegilops tauschii from Dx5 of Common Wheat

Abstract Objective Due to the allergic nature of the pollen of Cryptomeria japonica, the most important Japanese forestry conifer, a pollen-free cultivar is preferred. Mutant trees detected in nature have been used to produce a pollen-free cultivar. In order to reduce the time and cost needed for production and breeding, we aimed to develop simple diagnostic molecular markers for mutant alleles of the causative gene MALE STERILITY 1 (MS1) in C. japonica to rapidly identify pollen-free mutants. Results We developed PCR and LAMP markers to detect mutant alleles and to present experimental options depending on available laboratory equipment. LAMP markers were developed for field stations, where PCR machines are unavailable. The LAMP method only needs heat-blocks or a water bath to perform the isothermal amplification and assay results can be read by the naked eye. Because the causative mutations were deletions, we developed two kinds of PCR markers, amplified length polymorphism (ALP) and allele specific PCR (ASP) markers. These assays can be visualized using capillary or agarose gel electrophoresis.

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Evaluating the genetic effects of seed dormancy regulatory genes Qsd1 and Qsd2 in a global collection of cultivated barley ( Hordeum vulgare ssp. vulgare ) with functional kompetitive allele‐specific PCR markers

Plant Breeding ◽

10.1111/pbr.12955 ◽

2021 ◽

Author(s):

Luxi Yan ◽

Congcong Jiang ◽

Yu Cai ◽

Yuhan Pan ◽

Rugen Xu ◽

...

Keyword(s):

Hordeum Vulgare ◽

Seed Dormancy ◽

Genetic Effects ◽

Regulatory Genes ◽

Pcr Markers ◽

Specific Pcr ◽

Allele Specific ◽

Cultivated Barley ◽

Allele Specific Pcr ◽

Hordeum Vulgare Ssp

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Interaction between Glu-A1 and Glu-D1 alleles in physical dough property in Japanese soft wheats and development of allele-specific PCR markers for Glu-A1

Breeding Research ◽

10.1270/jsbbr.10.41 ◽

2008 ◽

Vol 10 (2) ◽

pp. 41-48 ◽

Cited By ~ 3

Author(s):

Kanenori Takata ◽

Mikiko Yanaka ◽

Tatsuya M. Ikeda ◽

Naoyuki Ishikawa

Keyword(s):

Pcr Markers ◽

Specific Pcr ◽

Allele Specific ◽

Allele Specific Pcr ◽

Dough Property

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Development of diagnostic PCR and LAMP markers for MALE STERILITY 1 (MS1) in Cryptomeria japonica D. Don

10.1101/2020.05.19.090092 ◽

2020 ◽

Author(s):

Yoichi Hasegawa ◽

Saneyoshi Ueno ◽

Fu-Jin Wei ◽

Asako Matsumoto ◽

Tokuko Ujino-Ihara ◽

...

Keyword(s):

Male Sterility ◽

Cryptomeria Japonica ◽

Agarose Gel Electrophoresis ◽

Pcr Markers ◽

Causative Gene ◽

Laboratory Equipment ◽

Diagnostic Pcr ◽

Specific Pcr ◽

Allele Specific ◽

Allele Specific Pcr

AbstractObjectiveDue to the allergic nature of the pollen of Cryptomeria japonica, the most important Japanese forestry conifer, a pollen-free cultivar is preferred. Mutant trees detected in nature have been used for the production of a pollen-free cultivar. In order to reduce the time and cost needed for the production and breeding, we aimed to develop simple diagnostic molecular markers for mutant alleles of the causative gene MALE STERILITY 1 (MS1) in C. japonica. The expected function of this gene, its two dysfunctional mutations, and genetic diversity were described recently in a related study.ResultsWe have developed PCR and LAMP markers to detect mutant alleles and to present experimental options depending on available laboratory equipment. At field stations, where PCR machines are not available, LAMP markers were developed. LAMP only needs heat-blocks or a water bath to perform the isothermal amplification and assay results can be easily seen by eye. Because the causative mutations were deletions, two kinds of PCR markers, amplified length polymorphism (ALP) and allele specific PCR (ASP) markers, were developed. These assays can be carried out by capillary or agarose gel electrophoresis.

Download Full-text