Mapping and expression profiling reveal an inserted fragment from purple mustard involved anthocyanin accumulation in Chinese cabbage
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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.
KeywordsMapping Expression analysis HPLC-MS Purple mustard fragment Chinese cabbage
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.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The authors declare that this study complies with the current laws of the countries in which the experiments were performed.
- Andersen OM, Markham KR (2005) Flavonoids: chemistry, biochemistry and applications. CRC Press, Taylor & Francis, Boca Raton. 397–398Google Scholar
- Edwards R, Dixon DP (2000) The role of glutathione transferases in herbicide metabolism. Herbic Mech Action 8:38–71Google Scholar
- Giusti MM, Wrolstad RE (2001) Characterization and measurement with UV–visible spectroscopy. Curr Protoc Food Anal Chem Unit F1(2):1–13Google Scholar
- Harborne JB, Baxter H (1993) Phytochemical dictionary. A handbook of bioactive compounds from plants, 2nd edn. Taylor & Francis Limited, CRC Press, Boca Raton, p 361–363Google Scholar
- Kosambi D (1943) The estimation of map distances from recombination values. Ann Hum Genet 12(1):172–175Google Scholar
- Liu X, Xiao G, Chen W, Xu Y, Wu J (2004) Quantification and purification of mulberry anthocyanins with macroporous resins. BioMed Res Int 2004:326–331Google Scholar
- Mazza G, Miniati E (1993) Anthocyanins in fruits, vegetables, and grains. CRC Press, Boca RatonGoogle Scholar
- 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 7:389–452Google Scholar
- Sun R, Zhang S, Zhang S, Li F (2006) Research on creation of purple Chinese cabbage germplasm. Acta Hortic Sin 33(5):1032Google Scholar
- 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):218–235CrossRefPubMedGoogle Scholar
- Van Ooijen J (2006) JoinMap 4. Software for the calculation of genetic linkage maps in experimental populations Kyazma BV, Wageningen, NetherlandsGoogle Scholar
- 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 159–168Google Scholar
- 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):1085–1092Google Scholar
- 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:14–17Google Scholar
- 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):0901–0906Google Scholar
- 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 18:25–29Google Scholar