Euphytica

, Volume 152, Issue 2, pp 225–234 | Cite as

Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE

  • Cheng Chang
  • Haiping Zhang
  • Jie Xu
  • Weihua Li
  • Guangtian Liu
  • Mingshan You
  • Baoyun Li
Article

Abstract

Puroindoline genes (Pina and Pinb) form the molecular basis of wheat grain hardness or texture. Variations in either gene are reported to be associated with grain hardness in wheat. Here, a modified denaturing PAGE was used to detect Pina and Pinb allelic variations in 102 common wheat cultivars and other species related to wheat. Two variations of Pina (Pina-D1b and Pina-D1p) and five variations of Pinb (null, Pinb-D1b, Pinb-D1u, Pinb-D1v and Pinb-D1w) were identified in common (T. aestivum) and spelt wheats (T. aestivum ssp. spelta). No allelic variation was found in Tibet semi-wild wheat (T. aestivum ssp. tibetanum), club wheat (T. compactum), or Aegilops  tauschii. Puroindoline genes were absent in wild␣emmer (T. turgidum var. dicoccoides). The sequencing results of the PCR fragments from Pina and Pinb (except for null type) indicated allelic variants carrying single base mutation, such as Pina-D1p, Pinb-D1u, Pinb-D1v and Pinb-D1w were novel types. Our results showed that the modified PAGE used in this study provided a satisfactory resolving power for identifying single nucleotide mutations; therefore, it is a practical and simple tool to study allelic variation of Pina and Pinb in wheat and related species.

Keywords

Common wheat Grain hardness Modified denaturing PAGE Puroindoline genes Single nucleotide mutant 

Abbreviations

ARMS

Amplification Refractory Mutation System

ASO

Allele Specific Oligonuceotide

ASP

Allelic Specific Primers

DGGE

Denaturing Gradient Gel Electrophoresis

SSCP

Single Strand Conformation Polymorphism

STS

Sequenced Tagged Site

SDS-PAGE

Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis

A-PAGE

Acidic Polyacrylamide Gel Electrophoresis

2-D PAGE

Two-dimensional Polyacrylamide Gel Electrophoresis

TBE

Tris-Boric acid-EDTA

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Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Cheng Chang
    • 1
    • 2
  • Haiping Zhang
    • 2
  • Jie Xu
    • 1
  • Weihua Li
    • 1
  • Guangtian Liu
    • 1
  • Mingshan You
    • 1
  • Baoyun Li
    • 1
  1. 1.Beijing Key Laboratory of Crop Genetic Improvement, Key Laboratory of Crop Genomics & Genetic Improvement, Ministry of Agriculture, the People’s Republic of China, College of Agronomy and BiotechnologyChina Agricultural University (CAU)BeijingP.R. China
  2. 2.Institute of Crop Sciences/National Wheat Improvement CenterChinese Academy of Agricultural Sciences (CAAS)BeijingP.R. China

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