Molecular Breeding

, Volume 25, Issue 1, pp 167–177

Molecular marker linked to a chromosome region regulating seed Zn accumulation in barley

  • Behzad Sadeghzadeh
  • Zed Rengel
  • Chengdao Li
  • Hua’an Yang


Zinc deficiency is a critical nutritional problem in soils, restricting yield and nutritional quality of barley (Hordeum vulgare L.). Some genotypes (Zn-efficient) can produce greater yield and accumulate more Zn in seed under Zn deficiency than standard (Zn-inefficient) genotypes. However, there is little information regarding the genetics of Zn uptake/accumulation and location of genes conferring Zn efficiency in barley. Selection through molecular markers for seed Zn accumulation might be an efficient complementary breeding tool in barley. With the aim of developing molecular markers for increased accumulation of Zn in seed, a population of 150 DH lines derived from a cross between Clipper (low-Zn-accumulator) and Sahara 3771 (high-Zn-accumulator) was screened in the field and glasshouse for seed Zn concentration and content. One dominant DNA polymorphism was detected using the microsatellite-anchored fragment length polymorphism (MFLP) technique. The candidate MFLP marker was isolated from the MFLP gel, re-amplified by PCR, cloned, sequenced, and converted into simple sequence-specific and PCR-based marker. This marker, located on the short arm of chromosome 2H, might be useful for the improvement of barley nutritional quality and productivity programs in Zn-deficient environments. However, high seed Zn alone can not replace the need for Zn fertilization.


MFLP Molecular marker Seed Zn accumulation Barley Zn efficiency 


  1. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1998) Current protocols in molecular biology. Wiley, New York, p 7.4A1-4Google Scholar
  2. Beavis WD, Keim P (1996) Identification of quantitative trait loci that are affected by environment. In: Kang MS, Gauch HG (eds) Genotype-by-environment interaction. CRC Press, Boca Raton, pp 123–149Google Scholar
  3. Boersma JG, Buirchell BJ, Sivasithamparam K, Yang H (2007a) Development of a sequence-specific PCR marker linked to the Ku gene which removes the vernalization requirement in narrow-leafed lupin. Plant Breeding 126:306–309CrossRefGoogle Scholar
  4. Boersma JG, Buirchell BJ, Sivasithamparam K, Yang H (2007b) Development of two sequence-specific PCR markers linked to the gene that reduces pod shattering in narrow-leafed Lupin (Lupinus ngustifolius L.). Genet Mol Biol 30:623–629CrossRefGoogle Scholar
  5. Bouis HE (2007) Micronutrient fortification of plants through plant breeding: can it improve nutrition in man at low cost? Proc Nutr Soc 62:403–411CrossRefGoogle Scholar
  6. Brugmans B, van der Hulst RGM, Visser RGF, Lindhout P, van Eck HJ, Journals O (2003) A new and versatile method for the successful conversion of AFLP TM markers into simple single locus markers. Nucleic Acids Res 31:e55CrossRefPubMedGoogle Scholar
  7. Cakmak I (2002) Plant nutrition research: priorities to meet human needs for food in sustainable ways. Plant Soil 247:3–24CrossRefGoogle Scholar
  8. Donini P, Stephenson P, Bryan GJ, Koebner RMD (1998) The potential of microsatellites for high throughput genetic diversity assessment in wheat and barley. Genet Resour Crop Evol 45:415–421CrossRefGoogle Scholar
  9. Ellis MH, Spielmeyer W, Gale KR, Rebetzke GJ, Richards RA (2002) “Perfect” markers for the Rht-B1b and Rht-D1b dwarfing genes in wheat. Theor Appl Genet 105:1038–1042CrossRefPubMedGoogle Scholar
  10. Filatov V, Dowdle J, Smirnoff N, Ford-Lloyd B, Newbury HJ, Macnair MR (2007) A quantitative trait loci analysis of zinc hyperaccumulation in Arabidopsis halleri. New Phytol 174:580–590CrossRefPubMedGoogle Scholar
  11. George MLC, Nelson RJ, Zeigler RS, Leung H (1998) Rapid population analysis of Magnaporthe grisea by using rep-PCR and endogenous repetitive DNA sequences. Phytopathology 88:223–229CrossRefPubMedGoogle Scholar
  12. Ghandilyan A, Vreugdenhil D, Aarts MGM (2006) Progress in the genetic understanding of plant iron and zinc nutrition. Physiol Plant 126:407–417CrossRefGoogle Scholar
  13. Graham RD, Senadhira D, Beebe S, Iglesias C, Monesterio I (1999) Breeding for micronutrient density in edible portions of staple food crops: conventional approaches. Field Crops Res 60:57–80CrossRefGoogle Scholar
  14. Gregorio GB, Senadhira D, Htut T, Graham RD (2000) Breeding for trace mineral density in rice. Food Nutr Bull 21:382–386Google Scholar
  15. Gupta PK, Varshney RK, Sharma PC, Ramesh B (1999) Molecular markers and their applications in wheat breeding. Plant Breeding 118:369–390CrossRefGoogle Scholar
  16. Guzman-Maldonado SH, Martinez O, Acosta-Gallegos JA, Guevara-Lara F, Paredes-Lopez O (2003) Putative quantitative trait loci for physical and chemical components of common bean. Crop Sci 43:1029–1035CrossRefGoogle Scholar
  17. Haymes KM (1996) Mini-prep method suitable for a plant breeding program. Plant Mol Biol 14:280–284CrossRefGoogle Scholar
  18. Islam AKMR, Shepherd KW (1981) Production of disomic wheat-barley chromosome addition lines using Hordeum bulbosum crosses. Genet Res 37:215–219CrossRefGoogle Scholar
  19. Issac RA, Kerber JD (1971) Techniques and uses in soil, plant and water analysis. In: Walsh LM (ed) Instrumental methods for analysis of soils and plant tissue. Soil Science Society of America, Madison, WI, pp 39–65Google Scholar
  20. Karakousis A, Barr AR, Kretschmer JM, Manning S, Jefferies SP, Chalmers KJ, Islam AKM, Langridge P (2003) Mapping and QTL analysis of the barley population Clipper × Sahara. Aust J Agric Res 54:1137–1140CrossRefGoogle Scholar
  21. Litt M, Luty JA (1989) A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. Am J Hum Genet 44:397–401PubMedGoogle Scholar
  22. Lonergan PF (2001) Genetic characterisation and QTL mapping of zinc nutrition in barley (Hordeum vulgare). PhD thesis, Faculty of Agriculture and Natural Resource Sciences. The University of Adelaide, AdelaideGoogle Scholar
  23. Longerich HP, Jenner GA, Fryer BJ, Jackson SE (1990) Inductively coupled plasma-mass spectrometric analysis of geological samples: a critical evaluation based on case studies. Chem Geol 83:105–118CrossRefGoogle Scholar
  24. Manly KF, Cudmore RH Jr, Meer JM (2001) MapManager QTX, cross-platform software for genetic mapping. Mamm Genome 12:930–932CrossRefPubMedGoogle Scholar
  25. Mantovi P, Bonazzi G, Maestri E, Marmiroli N (2003) Accumulation of copper and zinc from liquid manure in agricultural soils and crop plants. Plant Soil 250:249–257CrossRefGoogle Scholar
  26. Moraghan JT, Grafton K (1999) Seed-zinc concentration and the zinc-efficiency trait in navy bean. Soil Sci Soc Am J 63:918–922Google Scholar
  27. Poletti S, Gruissem W, Sautter C (2004) The nutritional fortification of cereals. Curr Opin Biotechnol 15:162–165CrossRefPubMedGoogle Scholar
  28. Raboy V, Dickinson DB, Below FE (1984) Variation in seed total phosphorus, phytic acid, zinc, calcium, magnesium, and protein among lines of Glycine max and G. soja. Crop Sci 24:431Google Scholar
  29. Sadeghzadeh B, Rengel Z, Li CD (2008) Mapping of chromosome regions associated with seed Zn accumulation in barley, PhD thesis. Faculty of Natural and Agricultural Sciences. The University of Western Australia, PerthGoogle Scholar
  30. Shan X, Blake TK, Talbert LE (1999) Conversion of AFLP markers to sequence-specific PCR markers in barley and wheat. Theor Appl Genet 98:1072–1078CrossRefGoogle Scholar
  31. Uauy C, Distelfeld A, Fahima T, Blechl A, Dubcovsky J (2006) A NAC gene regulating senescence, improves grain protein, zinc, and iron content in wheat. Science 314:1298–1301CrossRefPubMedGoogle Scholar
  32. Vos P, Hogers R, Bleeker M, Reijans M, Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414CrossRefPubMedGoogle Scholar
  33. Vreugdenhil D, Aarts MGM, Koornneef M, Nelissen H, Ernst WHO (2004) Natural variation and QTL analysis for cationic mineral content in seeds of Arabidopsis thaliana. Plant Cell Environ 27:828–839CrossRefGoogle Scholar
  34. Weeden NF, Hemmatt M, Lawson DM, Lodhi M, Bell RL, Manganaris AG, Reischs BI, Brown SK, Ye GN (1994) Development and application of molecular marker linkage maps in woody fruit crops. Euphytica 77:71–75CrossRefGoogle Scholar
  35. Welch RM (1999) Importance of seed mineral nutrient reserves in crop growth and development. In: Rengel Z (ed) Mineral nutrition of crops: fundamental mechanisms and implications. Food Products Press, New York, pp 205–226Google Scholar
  36. Welch RM, Graham RD (2004) Breeding for micronutrients in staple food crops from a human nutrition perspective. J Exp Bot 55:353–364CrossRefPubMedGoogle Scholar
  37. Wissuwa M, Ismail AM, Yanagihara S (2006) Effects of zinc deficiency on rice growth and genetic factors contributing to tolerance. Plant Physiol 142:731–741CrossRefPubMedGoogle Scholar
  38. Yang H, Sweetingham MW, Cowling WA, Smith PMC (2001) DNA fingerprinting based on microsatellite-anchored fragment length polymorphisms, and isolation of sequence-specific PCR markers in lupin (Lupinus angustifolius L.). Mol Breeding 7:203–209CrossRefGoogle Scholar
  39. Yang H, Shankar M, Buirchell BJ, Sweetingham MW, Caminero C, Smith PMC (2002) Development of molecular markers using MFLP linked to a gene conferring resistance to Diaporthe toxica in narrow-leafed lupin (Lupinus angustifolius L.). Theor Appl Genet 105:265–270CrossRefPubMedGoogle Scholar
  40. Yang H, Boersma JG, You M, Buirchell BJ, Sweetingham MW (2004) Development and implementation of a sequence-specific PCR marker linked to a gene conferring resistance to anthracnose disease in narrow-leafed lupin (Lupinus angustifolius L.). Mol Breeding 14:145–151CrossRefGoogle Scholar
  41. You M, Boersma JG, Buirchell BJ, Sweetingham MW, Siddique KHM, Yang H (2005) A PCR-based molecular marker applicable for marker-assisted selection for anthracnose disease resistance in lupin breeding. Cell Mol Biol Lett 10:123–134PubMedGoogle Scholar
  42. Zimmerman M, Hurrel R (2002) Improving iron, zinc and vitamin A nutrition through plant biotechnology. Curr Opin Biotechnol 13:142–145CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Behzad Sadeghzadeh
    • 1
    • 2
  • Zed Rengel
    • 1
  • Chengdao Li
    • 1
    • 3
  • Hua’an Yang
    • 3
  1. 1.Soil Science and Plant Nutrition, Faculty of Natural and Agricultural SciencesUniversity of Western AustraliaCrawleyAustralia
  2. 2.Dryland Agricultural Research Institute (DARI)MaraghehIran
  3. 3.Department of Agriculture & FoodSouth PerthAustralia

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