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Molecular characterization and marker development for hexaploid wheat-specific HMW glutenin subunit 1By18 gene

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Abstract

Wheat grain high molecular weight glutenin subunits (HMW-GS) are the major determinants of dough elasticity and viscosity, and thus of breadmaking quality. Most known HMW-GS genes in bread wheat have been already cloned except for very few such as 1By18 and 1By19. In this study, HMW-GS 1By18 gene was isolated from bread wheat for the first time. In the past, this subunit has been considered to have positive effect on gluten quality. Molecular characterization of the 1By18 subunit gene showed that its complete coding nucleotide sequence was 2163 bp long, being highly identical to 1By8, with only three single nucleotide polymorphisms (SNPs) at sites 231, 233 and 1976 of the coding sequence. The deduced amino acid sequence of 1By18 exhibited two substitutions compared to 1By8. The deduced protein was further verified by expression in E. coli and LC–MS/MS analysis. Phylogenetic analysis showed that 1By18 was more closely related to 1By8, 1By9 and 1By16 than to 1By15 and 1By20. Molecular phylogenetic analysis indicated that the Glu-B1i allele emerged after the formation of modern hexaploid wheat. Two sets of SNP-based AS-PCR markers for 1By18 gene were developed and validated on 110 bread wheat varieties with different Glu-Bly alleles, and five recombinant inbred lines. Both markers could effectively distinguish 1By18 from the other genes at the Glu-B1 locus, thus making them valuable for marker-assisted selection in wheat quality improvement programs.

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Abbreviations

AS-PCR:

Allele-specific polymerase chain reaction

HMW-GS:

High molecular weight glutenin subunits

LC–MS/MS:

Liquid chromatography–tandem mass spectrometry

LMW-GS:

Low molecular weight glutenin subunit

MALDI-TOF-MS:

Matrix-assisted laser desorption ionization tandem time of flight mass spectrometry

MW:

Molecular weight

MYA:

Million years ago

ORF:

Open reading frame

RILS:

Recombination inbred lines

RP-HPLC:

Reversed-phase high-performance liquid chromatography

RP-UPLC:

Reversed-phase ultra-performance liquid chromatography

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SNP:

Single nucleotide polymorphism

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Acknowledgments

This research was financially supported by Grants from the National Natural Science Foundation of China (31471485), the National Key Project for Transgenic Crops in China (2014ZX08009-003), Natural Science Foundation of Beijing City and the Key Developmental Project of Science Technology, Beijing Municipal Commission of Education (KZ201410028031), and International Science and Technology Cooperation Program of China (2013DFG30530).

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    Authors and Affiliations

    1. College of Life Science, Capital Normal University, Beijing, 100048, China

      Xiaona Liang, Shoumin Zhen, Caixia Han, Chang Wang, Xiaohui Li & Yueming Yan

    2. Australia-China Wheat Improvement Centre, Murdoch University, and Australian Export Grains Innovation Centre, Perth, WA, 6150, Australia

      Wujun Ma

    3. Hubei Collaborative Innovation Center for Grain Industry, Jingzhou, 434025, China

      Yueming Yan

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    1. Xiaona Liang
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    Correspondence to Wujun Ma or Yueming Yan.

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    Xiaona Liang and Shoumin Zhen have contributed equally to this work.

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    Liang, X., Zhen, S., Han, C. et al. Molecular characterization and marker development for hexaploid wheat-specific HMW glutenin subunit 1By18 gene. Mol Breeding 35, 221 (2015). https://doi.org/10.1007/s11032-015-0406-2

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