Advertisement

Molecular Breeding

, Volume 30, Issue 3, pp 1371–1378 | Cite as

A novel molecular marker for the polyphenol oxidase gene located on chromosome 2B in common wheat

  • Hongqi Si
  • Zhiliang Zhou
  • Xiaobo Wang
  • Chuanxi Ma
Article

Abstract

Polyphenol oxidase (PPO) is a major cause of time-dependent darkening and discoloration in Asian noodles and other wheat-based products. One of the best ways to reduce this undesirable darkening is to breed new wheat cultivars with low PPO activity using efficient and reliable markers. Based on the sequence of a PPO gene SSPPO-B1 (GenBank accession no. AB254804) located on chromosome 2B of common wheat, 26 pairs of primers were designed to detect polymorphisms between wheat cultivars with low and high PPO activity. F-8, one of these primer pairs, amplified double fragments (band “a” of approximately 400 bp and band “b” of approximately 600 bp) in the cultivars with low PPO activity, and a single fragment (only band a) in the cultivars with high PPO activity. The differences between the fragments a and b include five indels and several single nucleotide polymorphisms, which occurred in intron II of the PPO gene. F-8 can be used as a sequence-tagged site marker to discriminate between two alleles Ppo-B1a (GQ303713) and Ppo-B1b (AB254804). The screening of 284 accessions of the core collection of Chinese wheat germplasms using the marker F-8 showed that the double fragments were present in 188 accessions, and the single fragments were present in the remaining 96 accessions. Statistical analysis revealed that the cultivars with the double fragments had significantly lower mean PPO activity than those with the single fragments. We also screened the 284 accessions using two additional markers, PPO18 for Ppo-A1 on chromosome 2A and STS01 for Ppo-D1 on chromosome 2D. Results showed that the combination of markers F-8, PPO18, and STS01 could reliably predict PPO activity. These markers can be used in wheat breeding programs for low PPO activity selection to improve the quality of wheat-based products.

Keywords

Triticum aestivum L. Polyphenol oxidase STS marker PPO gene 

Notes

Acknowledgments

Our work was supported by grants from the Provincial Natural Science Key Research Project of Anhui Colleges (KJ2011A113), the National Key Technologies R&D Program (2011BAD35B03), and China Agriculture Research System (CARS-03). We are very grateful to the State Key Laboratory of Crop Genetics & Germplasm Enhancement (Nanjing Agricultural University, Nanjing, China) for assistance with the marker chromosomal location experiment.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11032_2012_9723_MOESM1_ESM.doc (47 kb)
Supplementary material 1 Supplementary Fig. 1 DNA sequence alignment of PPO alleles with fragments a and b amplified using the primer pair F-8 from the cultivars with low PPO activity (b4, b5, b6, b10, b11, b12) and high PPO activity (a2, a7, a8, a9). The numbers represent the same cultivars as shown in Table 1. The fragment b1 and a3 did not provide accurate sequences because of the presence of nested peaks in their sequencing profiles. GQ373013, AB254804, EF070147, EF070148, EF070149, and EF070150 indicate Ppo-B1a, Ppo-B1b, Ppo-A1a, Ppo-A1b, Ppo-D1a, and Ppo-D1b, respectively. Intron II is underlined. The five indels are shaded in different colors. The sequence GTACCGCCAGGC that participated in the alternative splicing (Sun et al. 2011) is boxed. The lowercase letters and dots indicate SNPs (DOC 47 kb)
11032_2012_9723_MOESM2_ESM.jpg (12 kb)
Supplementary material 2 (JPG 13 kb)
11032_2012_9723_MOESM3_ESM.doc (44 kb)
Supplementary material 3 (DOC 43 kb)
11032_2012_9723_MOESM4_ESM.xls (56 kb)
Supplementary material 4 (XLS 56 kb)
11032_2012_9723_MOESM5_ESM.doc (32 kb)
Supplementary material 5 (DOC 32 kb)

References

  1. Anderson JV, Morris CF (2001) An improved whole-seed assay for screening wheat germplasm for polyphenol oxidase activity. Crop Sci 41:1697–1705CrossRefGoogle Scholar
  2. Anderson JV, Morris CF (2003) Purification and analysis of wheat grain polyphenol oxidase (PPO) protein. Cereal Chem 80:135–143CrossRefGoogle Scholar
  3. Anderson JV, Fuerst EP, Hurkman WJ, Vensel WH, Morris CF (2006) Biochemical and genetic characterization of wheat (Triticum spp.) kernel polyphenol oxidases. J Cereal Sci 44:353–367CrossRefGoogle Scholar
  4. Baik BK, Czuchajowska Z, Pomeranz Y (1995) Discoloration of dough for oriental noodles. Cereal Chem 72:198–205Google Scholar
  5. Beecher B, Skinner DZ (2011) Molecular cloning and expression analysis of multiple polyphenol oxidase genes in developing wheat (Triticum aestivum) kernels. J Cereal Sci 53:371–378CrossRefGoogle Scholar
  6. Bhattacharya M, Luo Q, Corke H (1999) Time-dependent changes in dough color in hexaploid wheat landraces differing in polyphenol oxidase activity. J Agric Food Chem 47:3579–3585PubMedCrossRefGoogle Scholar
  7. Chang C, Zhang HP, Xu J, You MS, Li BY, Liu GT (2007) Variation in two PPO genes associated with polyphenol oxidase activity in seeds of common wheat. Euphytica 154:181–193CrossRefGoogle Scholar
  8. Demeke T, Morris CF, Campbell KG, King GE, Anderson JA, Chang HG (2001) Wheat polyphenol oxidase: distribution and genetic mapping in three inbred line populations. Crop Sci 41:1750–1757CrossRefGoogle Scholar
  9. Feillet P, Autran JC, Icard-Vernire C (2000) Pasta brownness: an assessment. J Cereal Sci 32:215–233CrossRefGoogle Scholar
  10. Fuerst EP, Xu SS, Beecher B (2008) Genetic characterization of kernel polyphenol oxidases in wheat and related species. J Cereal Sci 48:359–368CrossRefGoogle Scholar
  11. Han J, Ma CX, Si HQ, Han LD, Chen J (2006) The genetic analysis and variation of PPO activity in the early generation of Yangmai 158 × Huaimai 18. J Triticeae Crops 26:38–41Google Scholar
  12. Hao CY, Dong YC, Wang LF et al (2008) Genetic diversity and construction of core collection in Chinese wheat genetic resources. Chin Sci Bull 53:1518–1526CrossRefGoogle Scholar
  13. Hatcher DW, Symons SJ, Kruger JE (1999) Measurement of the time-dependent appearance of discolored spots in alkaline noodles by image analysis. Cereal Chem 76:189–194CrossRefGoogle Scholar
  14. He XY, He ZH, Zhang LP et al (2007) Allelic variation of polyphenol oxidase (PPO) genes located on chromosomes 2A and 2D and development of functional markers for the PPO genes in common wheat. Theor Appl Genet 115:47–58PubMedCrossRefGoogle Scholar
  15. He XY, He ZH, Morris CF, Xia XC (2009) Cloning and phylogenetic analysis of polyphenol oxidase genes in common wheat and related species. Genet Resour Crop Evol 56:311–321CrossRefGoogle Scholar
  16. Jimenez M, Dubcovsky J (1999) Chromosome location of genes affecting polyphenol oxidase activity in seeds of common and durum wheat. Plant Breed 118:395–398CrossRefGoogle Scholar
  17. Jukanti AK, Bruckner PL, Fischer AM (2004) Evaluation of wheat polyphenol oxidase genes. Cereal Chem 81:481–485CrossRefGoogle Scholar
  18. Kang HW, Cho YG, Yoon UH, Eun MY (1998) A rapid DNA extraction method for RFLP and PCR analysis from a single dry seed. Plant Mol Biol Rep 16:1–9CrossRefGoogle Scholar
  19. Kruger JE, Matsuo RR, Preston K (1992) A comparison of methods for the prediction of Cantonese noodle colour. Can J Plant Sci 72:1021–1029CrossRefGoogle Scholar
  20. Kruger JE, Anderson MH, Dexter JE (1994a) Effect of flour refinement on raw cantonese noodle color and texture. Cereal Chem 71:177–182Google Scholar
  21. Kruger JE, Hatcher DW, Pauw RD (1994b) A whole seed assay for polyphenol oxidase in Canadian prairie spring wheats and its usefulness as a measure of noodle darkening. Cereal Chem 71:324–326Google Scholar
  22. Massa AN, Beecher B, Morris CF (2007) Polyphenol oxidase (PPO) in wheat and wild relatives: molecular evidence for a multigene family. Theor Appl Genet 114:1239–1247PubMedCrossRefGoogle Scholar
  23. Morris CF, Jeffers HC, Engle DA (2000) Effect of processing, formula and measurement variables on alkaline noodle color-toward an optimized laboratory system. Cereal Chem 77:77–85CrossRefGoogle Scholar
  24. Nagai T, Suzuki N (2001) Partial purification of polyphenol oxidase from Chinese cabbage Brassica rapa L. J Agric Food Chem 49:3922–3926PubMedCrossRefGoogle Scholar
  25. Okot-Kotber M, Liavoga A, Yong KJ, Bagorogoza K (2002) Activation of polyphenol oxidase in extracts of bran from several wheat (Triticum aestivum) cultivars using organic solvents, detergents, and chaotropes. J Agric Food Chem 50:2410–2417PubMedCrossRefGoogle Scholar
  26. Park WJ, Shelton DR, Peterson CJ, Martin TJ, Kachman SD, Wehling RL (1997) Variation in polyphenol oxidase activity and quality characteristics among hard white wheat and hard red winter wheat samples. Cereal Chem 74:7–11CrossRefGoogle Scholar
  27. Raman R, Raman H, Johnstone K, Lisle C, Smith A, Martin P, Allen H (2005) Genetic and in silico comparative mapping of the polyphenol oxidase gene in bread wheat (Triticum aestivum L.). Funct Integr Genomic 5:185–200CrossRefGoogle Scholar
  28. Raman R, Raman H, Martin P (2007) Functional gene markers for polyphenol oxidase locus in bread wheat (Triticum aestivum L.). Mol Breed 19:315–328CrossRefGoogle Scholar
  29. Sun DJ, He ZH, Xia XC et al (2005) A novel STS marker for polyphenol oxidase activity in bread wheat. Mol Breed 16:209–218CrossRefGoogle Scholar
  30. Sun YW, He ZH, Ma WJ, Xia XC (2011) Alternative splicing in the coding region of ppo-a1 directly influences the polyphenol oxidase activity in common wheat (Triticum aestivum L.). Funct Integr Genomics 11:85–93PubMedCrossRefGoogle Scholar
  31. Wang XB, Ma CX, He KQ, Si HQ, Zhang YL (2008) Development and application of a STS marker for grain PPO gene located on chromosome 2D in common wheat. Sci Agric Sin 39:1583–1590Google Scholar
  32. Wang XB, Ma CX, Si HQ et al (2009) Gene markers for grain polyphenol oxidase activity in common wheat. Mol Breed 23:163–170CrossRefGoogle Scholar
  33. Watanabe N, Takeuchi A, Nakayama A (2004) Inheritance and chromosomal location of the homeologous genes affecting phenol colour reaction of kernels in durum wheat. Euphytica 139:87–93CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Hongqi Si
    • 1
    • 2
    • 3
  • Zhiliang Zhou
    • 1
  • Xiaobo Wang
    • 1
  • Chuanxi Ma
    • 1
    • 2
    • 3
  1. 1.School of AgronomyAnhui Agricultural UniversityHefeiChina
  2. 2.Key Laboratory of Wheat Biology and Genetic Breeding in Southern Huanghuai Wheat RegionMinistry of AgricultureHefeiChina
  3. 3.Anhui Key Laboratory of Crop BiologyHefeiChina

Personalised recommendations