, Volume 207, Issue 2, pp 377–385 | Cite as

Mapping of Pi, a gene conferring pink leaf in ornamental kale (Brassica oleracea L. var. acephala DC)

  • Pengfang ZhuEmail author
  • Mingming Cheng
  • Xin Feng
  • Yong Xiong
  • Chang Liu
  • Yaohai Kang


We previously found that pink leaf is conferred by a single semi-dominant gene in ornamental kale. To map this gene, we constructed an F2 segregating population containing 184 individuals by crossing W02–7, a white-leaved inbred line, with P02–9, a pink-leaved inbred line. We screened 297 simple sequence repeat (SSR) and 437 sequence-related amplified polymorphism (SRAP) markers using bulked segregant analysis. We identified one SSR and seven SRAP markers that linked tightly to the pink-leaf (Pi) gene. To obtain more specific genomic markers, we converted six SRAPs to sequence-characterized amplified region (SCAR) markers. We constructed a genetic linkage map of the Pi locus using one SSR and five SCAR markers, spanning a total interval of 15.0 cM. One SSR marker, Ni2C12, and one co-dominant SCAR marker, Boac04, flanked Pi on either side at distances of 0.6 and 2.4 cM, respectively. Based on the reference genome sequence of Brassica oleracea, we positioned Pi on the top of chromosome C3. We also detected that pink hypocotyl co-segregated with pink inner leaf. These markers provide the basis for fine-scale mapping and cloning of Pi, and may be used for marker-assisted selection in ornamental kale breeding programs.


Brassica oleracea L.var. acephala DC Pink leaf Genetic mapping SSR SCAR 



We thank Professor Zhongyun Piao (College of Horticulture, Shenyang Agricultural University) for his kindly help in database analysis. This study was funded by National Natural Science Foundation of China (# 31101566).

Author contribution

P Zhu developed molecular markers, carried out sequence analysis, and wrote draft manuscript. M Cheng and Y Xiong carried out the phenotyping and genotyping steps, developed SCAR and morphological markers, and selected recombinants. X Feng and C Liu developed SSR markers and constructed genetic map. Y Kang developed SRAP and SCAR markers. All authors have read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Brown AF, Yousef GG, Chebrolu KK, Byrd RW, Everhart KW, Thomas A, Reid RW, Parkin IAP, Sharpe AG, Guzman I, Jackson EW (2014) High density single nucleotide polymorphism (SNP) array mapping in Brassica oleracea: identification of QTL associated with carotenoid variation in broccoli florets. Theor Appl Genet 127:2051–2064CrossRefGoogle Scholar
  2. Chiu L, Zhou X, Sarah B, Wu X, Ronald LP, Li L (2010) The purple cauliflower arises from activation of a MYB transcription factor. Plant Physiol 154:1470–1480CrossRefGoogle Scholar
  3. Crisp P, Walkey DGA, Bellman E, Roberts E (1975) A mutation affecting curd colour in cauliflower (Brassica oleracea L. var. botrytis DC). Euphytica 24:173–176CrossRefGoogle Scholar
  4. Feng H, Li Y, Liu Z, Liu J (2012) Mapping of or, a gene conferring orange color on the inner leaf of the Chinese cabbage (Brassica rapa L. ssp. pekinensis). Mol Breed 29:235–244CrossRefGoogle Scholar
  5. Horejsi T, Box MJ, Staub EJ (1999) Efficiency of randomly amplified polymorphic DNA to sequence characterized amplified region marker conversion and their comparative polymerase chain reaction sensitivity in cucumber. J Am Soc Hort Sci. 124:128–135Google Scholar
  6. Kaga A, Ohnishi M, Ishii T, Kamijima O (1996) A genetic linkage map of azuki bean constructed with molecular and morphological markers using an interspecific population (Vigna angularis × V. nakashimae). Theor Appl Genet 93:658–663CrossRefGoogle Scholar
  7. Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10:236–242CrossRefGoogle Scholar
  8. Kosambi DD (1943) The estimation of map distances from recombination values. Ann Eugen 12:172–175CrossRefGoogle Scholar
  9. Li L, Garvin DF (2003) Molecular mapping of Or, a gene inducing β-carotene accumulation in cauliflower (Brassica oleracea var. botrytis). Genome 47:588–594CrossRefGoogle Scholar
  10. Li G, Quiros CF (2001) Sequence-related amplified polymorphism (SRAP), a new marker system based on a simple PCR reaction: its application to mapping and gene tagging in Brassica. Theor Appl Genet 103:455–461CrossRefGoogle Scholar
  11. Li L, Paolillo DJ, Parthasarathy MV, DiMuzio EM, Garvin DF (2001) A novel gene mutation that confers abnormal patterns of β-carotene accumulation in cauliflower (Brassica oleracea var. botrytis). Plant J 26:59–67CrossRefGoogle Scholar
  12. Li L, Lu S, Halloran D, Garvin D (2003) High-resolution genetic and physical mapping of the cauliflower high-β-carotene gene Or (Orange). Mol Genet Genomics 270:132–138CrossRefGoogle Scholar
  13. Liu R, Meng J (2003) MapDraw: a microsoft excel macro for drawing genetic linkage maps based on given genetic linkage data. Hereditas 25:317–321PubMedGoogle Scholar
  14. Liu S, Liu Y, Yang X, Tong C, Edwards D, Parkin IAP et al (2014) The Brassica oleracea genome reveals the asymmetrical evolution of polyploid genomes. Nat Commun 5:3930CrossRefGoogle Scholar
  15. Lu S, Joyce VE, Zhou XJ, Alex BL, Diana MO, Kelly MC, Brian JC, Dominick JP, David FG, Julia V, Leon VK, Hendrik K, Elizabeth DE, Cao J, Li L (2006) The cauliflower Or gene encodes a DnaJ cysteine-rich domaincontaining protein that mediates high levels of β-carotene accumulation. Plant Cell 18:3594–3605CrossRefGoogle Scholar
  16. Martin C, Gerats T (1993) Control of pigment biosynthesis genes during petal development. Plant Cell 5:1253–1264CrossRefGoogle Scholar
  17. Michelmore RW, Paran I, Kesseli RV (1991) Identification of marker linked to disease-resistance gene by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832CrossRefGoogle Scholar
  18. Rieseberg LH, Ellstrand NC (1993) What can molecular and morphological markers tell us about plant hybridization. Crit Rev Plant Sci 12:213–241Google Scholar
  19. Schneeberger K, Ossowski S, Lanz C, Juul T, Petersen AH, Nielsen KL, Jørgensen J, Weigel D, Andersen SU (2009) SHOREmap: simultaneous mapping and mutation identification by deep sequencing. Nat Methods 6:550–551CrossRefGoogle Scholar
  20. Van Ooijen JW (2006) Joinmap 4.0®, Software for the calculation of genetic linkage maps in experimental populations. Kyazma BV, WageningenGoogle Scholar
  21. Vidal JR, Delavault P, Coarer M, Defontaine A (2000) Design of grapevine (Vitis vinifera L.) cultivar-specific SCAR primers for PCR fingerprinting. Theor Appl Genet 101:1194–1201CrossRefGoogle Scholar
  22. Wang G, Zhang F, Yu Y, Zhang D, Zhao X (2007) Identification of SCAR markers linked to orange head leaf gene in Chinese cabbage (Brassica rapa L. ssp. pekinensis). Acta Horti Sin 34:217–220Google Scholar
  23. Wang Y, Liu Z, Li Y, Zhang Y, Yang X, Feng H (2013) Identification of sequence-related amplified polymorphism (SRAP) markers linked to the red leaf trait in ornamental kale (Brassica oleracea L. var. acephala). Genet Mol Res 12:870–877CrossRefGoogle Scholar
  24. Yan Z, Denneboom C, Hattendorf A, Dolstra O, Debener T, Stam P, Visser PB (2005) Construction of an integrated map of rose with AFLP, SSR, PK, RGA, RFLP, SCAR and morphological markers. Theor Appl Genet 110:766–777CrossRefGoogle Scholar
  25. Yu Y, Zhang Y, Zhang D (2009) SRAP markers linked to purple trait in Chinese cabbage. Mol Plant Breed 7:573–578CrossRefGoogle Scholar
  26. Zhang F, Wang G, Wang M, Liu X, Zhao X, Yu Y, Zhang D, Yu S (2008) Identification of SCAR markers linked to or, a gene inducing beta-carotene accumulation in Chinese cabbage. Euphytica 164:463–471CrossRefGoogle Scholar
  27. Zhang J, Li H, Zhang M, Hui M, Wang Q, Li L, Zhang L (2013) Fine mapping and identification of candidate Br-or gene controlling orange head of Chinese cabbage (Brassica rapa L. ssp. pekinensis). Mol Breeding 32:799–805CrossRefGoogle Scholar
  28. Zhu P, Wei Y (2009) Compatibility, production of interspecific F1 and BC1 between improved CMS Brassica campestris ssp. pekinensis and B. oleracea var. acephala. J Plant Breed Crop Sci 1:265–269Google Scholar
  29. Zhu P, Zhang J, Fang X, Wang X, Huang J (2012) Heredity analyses of pink leaf in Brassica oleracea var. acephala. Adv Ornam Hortic China 1:177–180 Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Pengfang Zhu
    • 1
    Email author
  • Mingming Cheng
    • 1
  • Xin Feng
    • 1
  • Yong Xiong
    • 1
  • Chang Liu
    • 1
  • Yaohai Kang
    • 1
  1. 1.College of ForestryShenyang Agricultural UniversityShenyangChina

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