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Efficient isolation of ion beam-induced mutants for homoeologous loci in common wheat and comparison of the contributions of Glu-1 loci to gluten functionality

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Abstract

Key message

Ion beam mutations can be efficiently isolated and deployed for functional comparison of homoeologous loci in polyploid plants, and Glu - 1 loci differ substantially in their contribution to wheat gluten functionality.

Abstract

To efficiently conduct genetic analysis, it is beneficial to have multiple types of mutants for the genes under investigation. Here, we demonstrate that ion beam-induced deletion mutants can be efficiently isolated for comparing the function of homoeologous loci of common wheat (Triticum aestivum). Through fragment analysis of PCR products from M2 plants, ion beam mutants lacking homoeologous Glu-A1, Glu-B1 or Glu-D1 loci, which encode high molecular weight glutenin subunits (HMW-GSs) and affect gluten functionality and end-use quality of common wheat, could be isolated simultaneously. Three deletion lines missing Glu-A1, Glu-B1 or Glu-D1 were developed from the original mutants, with the Glu-1 genomic regions deleted in these lines estimated using newly developed DNA markers. Apart from lacking the target HMW-GSs, the three lines all showed decreased accumulation of low molecular weight glutenin subunits (LMW-GSs) and increased amounts of gliadins. Based on the test data of five gluten and glutenin macropolymer (GMP) parameters obtained with grain samples harvested from two environments, we conclude that the genetic effects of Glu-1 loci on gluten functionality can be ranked as Glu-D1 > Glu-B1 > Glu-A1. Furthermore, it is suggested that Glu-1 loci contribute to gluten functionality both directly (by promoting the formation of GMP) and indirectly (through keeping the balance among HMW-GSs, LMW-GSs and gliadins). Finally, the efficient isolation of ion beam mutations for functional comparison of homoeologous loci in polyploid plants and the usefulness of Glu-1 deletion lines for further studying the contribution of Glu-1 loci to gluten functionality are discussed.

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Abbreviations

EST:

Expressed sequence tag

FPC:

Flour protein content

G′:

Elastic modulus

G″:

Viscous modulus

Gli/Glu:

Gliadins/glutenins

GMP:

Glutenin macropolymer

HMW-GS:

High molecular weight glutenin subunit

LMW-GS:

Low molecular weight glutenin subunit

NILs:

Near isogenic lines

RP-HPLC:

Reverse-phase high-performance liquid chromatography

SDS:

Sodium dodecyl sulfate

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SDS-SV:

SDS-sedimentation volume

SIG:

Swelling index of glutenin

STS:

Sequence-tagged site

TGW:

Thousand grain weight

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Acknowledgments

This work was supported by grants from the Ministry of Science and Technology of China (2011BAD07B02-2 and 2009CB118300). We thank Professors Aimin Zhang and Jiazhu Sun for advice on measuring SDS-SV and SIG.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

The authors declare that the experiments described in this work were all conducted in compliance with the current laws of the Chinese government.

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Authors and Affiliations

  1. The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China

    Yushuang Yang, Shiming Li, Kunpu Zhang, Zhenying Dong, Yiwen Li, Xueli An, Jing Chen, Xin Liu, Huanju Qin & Daowen Wang

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Yushuang Yang, Shiming Li & Jing Chen

  3. Henan Provincial Key Laboratory of Ion Beam Bioengineering, Zhengzhou University, Zhengzhou, 450052, China

    Qiufang Chen & Zhen Jiao

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  1. Yushuang Yang
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  2. Shiming Li
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  3. Kunpu Zhang
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  10. Xin Liu
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Correspondence to Daowen Wang.

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Yang, Y., Li, S., Zhang, K. et al. Efficient isolation of ion beam-induced mutants for homoeologous loci in common wheat and comparison of the contributions of Glu-1 loci to gluten functionality. Theor Appl Genet 127, 359–372 (2014). https://doi.org/10.1007/s00122-013-2224-4

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