Abstract
Anthocyanins are the major pigments responsible for purple coloration in flowers, fruits and leaves, and the genes involved in their biosynthetic pathway have been identified in many plants. A purple-leaf Chinese cabbage (Brassica rapa L. ssp. pekinensis) was bred by interspecies crossing between Chinese cabbage and purple-leaf mustard [Brassica juncea (L.) Coss. var. foliosa L. H. Bailey]. In this study, high-performance liquid chromatographic analysis indicated purple coloration in Chinese cabbage is due to the accumulation of the same kind of cyaninin as in purple mustard. To elucidate the genetic factors controlling anthocyanin accumulation in this purple-leaf Chinese cabbage, we mapped the anthocyanin gene from the mustard (Anm) locus in an F2 population and performed expression profiling of anthocyanin-related genes. A genetic analysis revealed that the purple-leaf phenotype is a qualitative trait and that its inheritance is unstable in purple-leaf Chinese cabbage. Mapping insertion/deletion markers from 288 individuals of the F2 population located the Anm locus within a 2.5-cM interval on B. rapa chromosome A02. The sequencing and alignment of the amplified fragments demonstrated that purple Chinese cabbage contains fragments of purple mustard on chromosome A02. We evaluated the expression profiles of 12 anthocyanin-related genes on A02 by reverse-transcription and quantitative real-time PCR methods, which revealed that the expression levels of five genes were higher in purple Chinese cabbage than in the non-purple variety. These results offer insights into the molecular mechanism of anthocyanin biosynthesis and improve the knowledge on molecular breeding of purple-type Chinese cabbage.
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References
Andersen OM, Markham KR (2005) Flavonoids: chemistry, biochemistry and applications. CRC Press, Taylor & Francis, Boca Raton. 397–398
Axelsson T, Bowman C, Sharpe A, Lydiate D, Lagercrantz U (2000) Amphidiploid Brassica juncea contains conserved progenitor genomes. Genome 43:679–688
Banga S (1988) C-genome chromosome substitution lines in Brassica juncea (L.) Coss. Genetica 77:81–84
Boss PK, Davies C, Robinson SP (1996) Expression of anthocyanin biosynthesis pathway genes in red and white grapes. Plant Mol Biol 32(3):565–569
Broun P (2005) Transcriptional control of flavonoid biosynthesis: a complex network of conserved regulators involved in multiple aspects of differentiation in Arabidopsis. Curr Opin Plant Biol 8(3):272–279
Chen DH, Ronald P (1999) A rapid DNA minipreparation method suitable for AFLP and other PCR applications. Plant Mol Biol Rep 17(1):53–57
Chiu LW, 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(3):1470–1480
De Jong W, Eannetta N, De Jong D, Bodis M (2004) Candidate gene analysis of anthocyanin pigmentation loci in the Solanaceae. Theor Appl Genet 108(3):423–432
Dixon RA, Xie DY, Sharma SB (2005) Proanthocyanidins–a final frontier in flavonoid research? New Phytol 165(1):9–28
Edwards R, Dixon DP (2000) The role of glutathione transferases in herbicide metabolism. Herbic Mech Action 8:38–71
Giusti MM, Wrolstad RE (2001) Characterization and measurement with UV–visible spectroscopy. Curr Protoc Food Anal Chem Unit F1(2):1–13
Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780
Guo N, Cheng F, Wu J, Liu B, Zheng S, Liang J, Wang X (2014) Anthocyanin biosynthetic genes in Brassica rapa. BMC Genom 15(1):426
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 Breed 35(5):1–11
Harborne JB, Baxter H (1993) Phytochemical dictionary. A handbook of bioactive compounds from plants, 2nd edn. Taylor & Francis Limited, CRC Press, Boca Raton, p 361–363
Hayashi K, Matsumoto S, Tsukazaki H, Kondo T, Kubo N, Hirai M (2010) Mapping of a novel locus regulating anthocyanin pigmentation in Brassica rapa. Breed Sci 60(1):76–80
Holton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7(7):1071–1083
Islam A, Shepherd K (1992) Production of wheat-barley recombinant chromosomes through induced homoeologous pairing. Theor Appl Genet 83(4):489–494
Jiang J, Friebe B, Gill BS (1993) Recent advances in alien gene transfer in wheat. Euphytica 73(3):199–212
Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10(5):236–242
Kosambi D (1943) The estimation of map distances from recombination values. Ann Hum Genet 12(1):172–175
Lam TK, Gallicchio L, Lindsley K, Shiels M, Hammond E, Tao XG, Chen L, Robinson KA, Caulfield LE, Herman JG (2009) Cruciferous vegetable consumption and lung cancer risk: a systematic review. Cancer Epidem Biomark 18(1):184–195
Langfelder P, Horvath S (2008) WGCNA: an R package for weighted correlation network analysis. BMC Bioinf 9(1):559
Lee Y, Yoon HR, Paik YS, Liu JR, W-i Chung, Choi G (2005) Reciprocal regulation of Arabidopsis UGT78D2 and BANYULS is critical for regulation of the metabolic flux of anthocyanidins to condensed tannins in developing seed coats. J Plant Biol 48(4):356–370
Li P, Zhang S, Zhang S, Li F, Zhang H, Cheng F, Wu J, Wang X, Sun R (2015a) Carotenoid biosynthetic genes in Brassica rapa: comparative genomic analysis, phylogenetic analysis, and expression profiling. BMC Genom 16(1):492
Li P, Zhang S, Zhang S, Li F, Zhang H, Liu X, Wu J, Wang X, Sun R (2015b) Carotenoid identification and molecular analysis of carotenoid isomerase-encoding BrCRTISO, the candidate gene for inner leaf orange coloration in Chinese cabbage. Mol Breed 35(2):1–12
Lin LZ, Sun J, Chen P, Harnly J (2011) UHPLC-PDA-ESI/HRMS/MSn analysis of anthocyanins, flavonol glycosides, and hydroxycinnamic acid derivatives in red mustard greens (Brassica juncea Coss Variety). J Agric Food Chem 59(22):12059–12072
Liu X, Xiao G, Chen W, Xu Y, Wu J (2004) Quantification and purification of mulberry anthocyanins with macroporous resins. BioMed Res Int 2004:326–331
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408
Marrs KA, Alfenito MR, Lloyd AM, Walbot V (1995) A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2. Nature 375:397–400
Mazza G, Miniati E (1993) Anthocyanins in fruits, vegetables, and grains. CRC Press, Boca Raton
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
Quiros C, Ochoa O, Kianian S, Douches D (1987) Analysis of the Brassica oleracea genome by the generation of B. campestris-oleracea chromosome addition lines: characterization by isozymes and rDNA genes. Theor Appl Genet 74(6):758–766
Sun R, Zhang S, Zhang S, Li F (2006) Research on creation of purple Chinese cabbage germplasm. Acta Hortic Sin 33(5):1032
Sun W, Meng X, Liang L, Jiang W, Huang Y, He J, Hu H, Almqvist J, Gao X, Wang L (2015) Molecular and biochemical analysis of chalcone synthase from Freesia hybrid in flavonoid biosynthetic pathway. PLoS One 10(3):e0119054
Tohge T, Nishiyama Y, Hirai MY, Yano M, Ji Nakajima, Awazuhara M, Inoue E, Takahashi H, Goodenowe DB, Kitayama M (2005) Functional genomics by integrated analysis of metabolome and transcriptome of Arabidopsis plants over-expressing an MYB transcription factor. Plant J 42(2):218–235
Van Ooijen J (2006) JoinMap 4. Software for the calculation of genetic linkage maps in experimental populations Kyazma BV, Wageningen, Netherlands
van Poppel G, Verhoeven DT, Verhagen H, Goldbohm RA (1999) Brassica vegetables and cancer prevention. In: Zappia V (ed) Advances in nutrition and cancer 2. Springer, New York, p 159–168
Winkel-Shirley B (2001a) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126(2):485–493
Winkel-Shirley B (2001b) It takes a garden. How work on diverse plant species has contributed to an understanding of flavonoid metabolism. Plant Physiol 127(4):1399–1404
Wu X, Prior RL (2005) Systematic identification and characterization of anthocyanins by HPLC-ESI-MS/MS in common foods in the United States: fruits and berries. J Agric Food Chem 53(7):2589–2599
Yamasaki H, Sakihama Y, Ikehara N (1997) Flavonoid-peroxidase reaction as a detoxification mechanism of plant cells against H2O2. Plant Physiol 115(4):1405–1412
Yan Z, Wang Y, Xuan S, Zhao J, Shen S (2015) Obtaining and genetic stability of Chinese cabbage—cabbage translocation lines with fragment of cabbage chromosome 8. Acta Hortic Sin 42(6):1085–1092
Yildiz M, Willis DK, Cavagnaro PF, Iorizzo M, Abak K, Simon PW (2013) Expression and mapping of anthocyanin biosynthesis genes in carrot. Theor Appl Genet 126(7):1689–1702
You Q, Wang B, Chen F, Huang Z, Wang X, Luo PG (2011) Comparison of anthocyanins and phenolics in organically and conventionally grown blueberries in selected cultivars. Food Chem 125:201–208
Zhang D, Zhang F, Yu Y, Zhao X, Yu S, Xu J (2008a) Initial study on breeding of purple Chinese cabbage. J Changjiang Veg 11b:14–17
Zhang M, Zhang L, Gong Z, Hui M (2008b) Screening RAPD markers linked to purple trait of Chinese cabbage and its chromosome location. Acta Bot Boreal Occident Sin 28(5):0901–0906
Zhang D, Wang W, Zhang F, Zhao X, Yu Y, Yu S, Xu J, Lu G (2011) Genetic relationship between Chinese cabbage with orange color in inner head and purple color in leaf. China Veg 18:25–29
Zhang B, Hu Z, Zhang Y, Li Y, Zhou S, Chen G (2012) A putative functional MYB transcription factor induced by low temperature regulates anthocyanin biosynthesis in purple kale (Brassica Oleracea var. acephala f. tricolor). Plant Cell Rep 31(2):281–289
Acknowledgments
This work was supported by a Chinese 973 Program Grant (2012CB113900) and a Chinese 863 Program grant (2012AA100100), both to RS. This study was also funded by the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-IVFCAAS). Research was carried out in the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, P. R. China.
Author contribution statement
SZ constructed the mapping populations and performed the genetic analysis and anthocyanin profile. PL performed mapping and expression analyses and wrote the paper. WQ extracted the DNA of the F2 population and provided advice on the manuscript. SFZ, FL, HZ and XW provided advice on experiments. RS designed and supervised the work. All the authors have read and approved the final manuscript.
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Shujiang Zhang and Peirong Li have equally contributed to this work and are regarded as joint first authors.
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Zhang, S., Li, P., Qian, W. et al. Mapping and expression profiling reveal an inserted fragment from purple mustard involved anthocyanin accumulation in Chinese cabbage. Euphytica 212, 83–95 (2016). https://doi.org/10.1007/s10681-016-1761-8
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DOI: https://doi.org/10.1007/s10681-016-1761-8