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Fine mapping and candidate gene analysis of an anthocyanin-rich gene, BnaA.PL1, conferring purple leaves in Brassica napus L.

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Abstract

Because of the advantages of anthocyanins, the genetics and breeding of crops rich in anthocyanins has become a hot research topic. However, due to the lack of anthocyanin-related mutants, no regulatory genes have been mapped in Brassica napus. In this study, we first report the characterization of a B. napus line with purple leaves and the fine mapping and candidate screening of the BnaA.PL1 gene. The amount of anthocyanins in the purple leaf line was six times higher than that in a green leaf line. A genetic analysis indicated that the purple character was controlled by an incomplete dominant gene. Through map-based cloning, we localized the BnaA.PL1 gene to a 99-kb region at the end of B. napus chromosome A03. Transcriptional analysis of 11 genes located in the target region revealed that the expression level of only the BnAPR2 gene in seedling leaves decreased from purple to reddish green to green individuals, a finding that was consistent with the measured anthocyanin accumulation levels. Molecular cloning and sequence analysis of BnAPR2 showed that the purple individual-derived allele contained 17 variants. Markers co-segregating with BnaA.PL1 were developed from the sequence of BnAPR2 and were validated in the BC4P2 population. These results suggested that BnAPR2, which encodes adenosine 5′-phosphosulfate reductase, is likely to be a valuable candidate gene. This work may lay the foundation for the marker-assisted selection of B. napus vegetables that are rich in anthocyanins and for an improved understanding of the molecular mechanisms controlling anthocyanin accumulation in Brassica.

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References

  1. Asem ID, Imotomba R, Mazumder P, Laishram J (2015) Anthocyanin content in the black scented rice (Chakhao): its impact on human health and plant defense. Symbiosis 66:47–54

  2. Berglund T, Ohlsson AB, RydstrÖm J (1993) Nicotinamide increases glutathione and anthocyanin in tissue culture of Catharanthus roseus. J Plant Physiol 141:596–600

  3. Burdzinski C, Wendell DL (2007) Mapping the anthocyaninless (anl) locus in rapid-cycling Brassica rapa (RBr) to linkage group R9. BMC Genet 8:64–69

  4. Chagné D, Bianco L, Lawley C, Micheletti D, Jacobs JM (2015) Methods for the design, implementation, and analysis of illumina Infinium™ SNP assays in plants. Plant Genotyping Methods Protoc 2015:281–298

  5. Chalhoub B, Denoeud F, Liu S, Parkin IA, Tang H, Wang X, Chiquet J, Belcram H, Tong C, Samans B (2014) Early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome. Science 345:950–953

  6. Chalker-Scott L (1999) Environmental significance of anthocyanins in plant stress responses. Photochem Photobiol 70:1–9

  7. Chao DY, Baraniecka P, Danku J, Koprivova A, Lahner B, Luo HB, Yakubova E, Dilkes B, Kopriva S, Salt DE (2014) Variation in sulfur and selenium accumulation is controlled by naturally occurring isoforms of the key sulfur assimilation enzyme adenosine 5′-phosphosulfate reductase 2 across the Arabidopsis species range. Plant Physiol 166:1593–1608

  8. Chiu L-W, Li L (2012) Characterization of the regulatory network of BoMYB2 in controlling anthocyanin biosynthesis in purple cauliflower. Planta 236:1153–1164

  9. Chiu L-W, Zhou X, Burke S, Wu X, Prior RL, Li L (2010) The purple cauliflower arises from activation of a MYB transcription factor. Plant Physiol 154:1470–1480

  10. Craker L, Wetherbee PJ (1973) Ethylene, light, and anthocyanin synthesis. Plant Physiol 51:436–438

  11. Dixon DP, Skipsey M, Edwards R (2010) Roles for glutathione transferases in plant secondary metabolism. Phytochemistry 71:338–350

  12. Doyle JJT, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

  13. El-Kereamy A, Chervin C, Roustan JP, Cheynier V, Souquet JM, Moutounet M, Raynal J, Ford C, Latché A, Pech JC (2003) Exogenous ethylene stimulates the long-term expression of genes related to anthocyanin biosynthesis in grape berries. Physiol Plantarum 119:175–182

  14. Fan G, Han Y, Gu Z, Chen D (2008) Optimizing conditions for anthocyanins extraction from purple sweet potato using response surface methodology (RSM). LWT Food Sci Technol 41:155–160

  15. Fujii K, Ohmido N (2011) Stable progeny production of the amphidiploid resynthesized Brassica napus cv. Hanakkori, a newly bred vegetable. Theor Appl Genet 123:1433–1443

  16. Grant K, Carry NM, Mendoza M, Schulze J, Pilon M, Pilon-Smits EAH, Hoewyk DV (2011) Adenosine 5-phosphosulfate reductase (APR2) mutation in Arabidopsis implicates glutathione deficiency in selenate toxicity. Biochem J 438:325–335

  17. Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780

  18. Guo N, Cheng F, Wu J, Liu B, Zheng S, Liang J, Wang X (2014) Anthocyanin biosynthetic genes in Brassica rapa. BMC Genom 15:426–436

  19. Guo N, Wu J, Zheng S, Cheng F, Liu B, Liang J, Cui Y, Wang X (2015) Anthocyanin profile characterization and quantitative trait locus mapping in zicaitai (Brassica rapa L. ssp. chinensis var. purpurea). Mol Breeding 35:1–11

  20. Hayashi K, Matsumoto S, Tsukazaki H, Kondo T, Kubo N, Hirai M (2010) Mapping of a novel locus regulating anthocyanin pigmentation in Brassica rapa. Breeding Sci 60:76–80

  21. Jiang C, Ramchiary N, Ma Y, Jin M, Feng J, Li R, Wang H, Long Y, Choi SR, Zhang C (2011) Structural and functional comparative mapping between the Brassica A genomes in allotetraploid Brassica napus and diploid Brassica rapa. Theor Appl Genet 123:927–941

  22. Kim C, Park S, Kikuchi S, Kwon S, Park S, Yoon U, Park D, Seol Y, Hahn J, Park S (2010) Genetic analysis of gene expression for pigmentation in Chinese cabbage (Brassica rapa). BioChip J 4:123–128

  23. Kim C, Kim J, Kikuchi S, Choi J, Kim Y, Park H, Seol Y, Park D, Hahn J, Kim Y (2011) Computational identification of Chinese cabbage anthocyanin specific genes. BioChip J 5:184–192

  24. Kortstee A, Khan S, Helderman C, Trindade L, Wu Y, Visser R, Brendolise C, Allan A, Schouten H, Jacobsen E (2011) Anthocyanin production as a potential visual selection marker during plant transformation. Transgenic Res 20:1253–1264

  25. Li X, Yu E, Fan C, Zhang C, Fu T, Zhou Y (2012) Developmental, cytological and transcriptional analysis of autotetraploid Arabidopsis. Planta 236:579–596

  26. Lin L-Z, Sun J, Chen P, Harnly J (2011) UHPLC-PDA-ESI/HRMS/MS n analysis of anthocyanins, flavonol glycosides, and hydroxycinnamic acid derivatives in red mustard greens (Brassica juncea Coss variety). J Agric Food Chem 59:12059–12072

  27. Lincoln SE, Daly MJ, Lander ES (1993) Constructing genetic linkage maps with MAPMAKER/EXP Version 3.0: a tutorial and reference manual. A Whitehead Institute for Biomedical Research Technical Report, 3rd edn

  28. Littell RC, Stroup WW, Milliken GA, Wolfinger RD, Schabenberger O (2006) SAS for mixed models. SAS Institute. Inc, Cary, p 814

  29. Liu J, Huang S, Sun M, Liu S, Liu Y, Wang W, Zhang X, Wang H, Hua W (2012) An improved allele-specific PCR primer design method for SNP marker analysis and its application. Plant Methods 8:34–42

  30. Lu G, Yang G, Fu T (2004) Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus). Plant Breeding 123:262–265

  31. Luo Y, Du D, Fu G, Xu L, Li X, Xing X, Yao Y, Zhang X, Zhao Z, Liu H (2011) Inheritance of leaf color and sequence-related amplified polymorphic (SRAP) molecular markers linked to the leaf color gene in Brassica juncea. AFR Jl Biotechnol 10:14724–14730

  32. Marrs KA (1996) The functions and regulation of glutathione S-transferases in plants. Annu Rev Plant Biol 47:127–158

  33. Mehrtens F, Kranz H, Bednarek P, Weisshaar B (2005) The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis. Plant Physiol 138:1083–1096

  34. Moons A (2005) Regulatory and functional interactions of plant growth regulators and plant glutathione S-transferases (GSTs). Vitam Horm 72:155–202

  35. Mueller LA, Goodman CD, Silady RA, Walbot V (2000) AN9, a petunia glutathione S-transferase required for anthocyanin sequestration, is a flavonoid-binding protein. Plant Physiol 123:1561–1570

  36. Nabavi S, Habtemariam S, Daglia M, Shafighi N, Barber A, Nabavi S (2015) Anthocyanins as a potential therapy for diabetic retinopathy. Curr Med Chem 22:51–58

  37. Nagaharu U (1935) Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization. Jpn J Bot 7:389–452

  38. Petroni K, Tonelli C (2011) Recent advances on the regulation of anthocyanin synthesis in reproductive organs. Plant Sci 181:219–229

  39. Piquemal J, Cinquin E, Couton F, Rondeau C, Seignoret E, Perret D, Villeger M-J, Vincourt P, Blanchard P (2005) Construction of an oilseed rape (Brassica napus L.) genetic map with SSR markers. Theor Appl Genet 111:1514–1523

  40. Podsędek A (2007) Natural antioxidants and antioxidant capacity of Brassica vegetables: a review. LWT Food Sci and Technol 40:1–11

  41. Vos P, Hogers R, Bleeker M, Reijans M, Van de Lee T, Hornes M, Friters A, Pot J, Paleman J, Kuiper M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

  42. Wang Y, Sun S, Liu B, Wang H, Deng J, Liao Y, Wang Q, Cheng F, Wang X, Wu J (2011) A sequence-based genetic linkage map as a reference for Brassica rapa pseudochromosome assembly. BMC Genom 12:239–247

  43. Wang W, Zhang D, Yu S, Liu J, Wang D, Zhang F, Yu Y, Zhao X, Lu G, Su T (2014) Mapping the BrPur gene for purple leaf color on linkage group A03 of Brassica rapa. Euphytica 199:293–302

  44. Yi B, Chen Y, Lei S, Tu J, Fu T (2006) Fine mapping of the recessive genic male-sterile gene (Bnms1) in Brassica napus L. Theor Appl Genet 113:643–650

  45. Yuan Y, Chiu L-W, Li L (2009) Transcriptional regulation of anthocyanin biosynthesis in red cabbage. Planta 230:1141–1153

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Acknowledgments

This research was financed by the Program of Introducing Talents of Discipline to Universities in China (the 111 Project No. B14032), the Program for Modern Agricultural Industrial Technology System (nycytx-00501), the National Support Program (2011BAD35B04), the Hi-Tech Research and Development Program of China (2011AA10A104), and the Science and Technology Projects of Shaoguan (2013CXY/C13, 2014CX/N323).

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Correspondence to Jing Wen.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Communicated by S. Hohmann.

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Li, H., Zhu, L., Yuan, G. et al. Fine mapping and candidate gene analysis of an anthocyanin-rich gene, BnaA.PL1, conferring purple leaves in Brassica napus L.. Mol Genet Genomics 291, 1523–1534 (2016). https://doi.org/10.1007/s00438-016-1199-7

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Keywords

  • Anthocyanin-rich mutant
  • Brassica napus
  • Fine mapping
  • Purple leaf trait