Metabolomic analysis reveals the interaction of primary and secondary metabolism in white, pale green, and green pak choi (Brassica rapa subsp. chinensis)

Hyeon Ji Yeo1, Seung‐A Baek2, Ramaraj Sathasivam1, Jae Kwang Kim2, Sang Un Park3
1Department of Crop Science, Chungnam National University, 99 Daehak-ro, Yuseong-gu, Daejeon 34134, Republic of Korea
2Division of Life Sciences and Bio-Resource and Environmental Center, Incheon National University, Yeonsu-gu, Incheon, 22012, Republic of Korea
3Department of Smart Agriculture Systems, Chungnam National University, 99 Daehak-Ro, Yuseong-gu, Daejeon 34134, Republic of Korea

Tóm tắt

Abstract

This study aimed to comprehensively analyze primary and secondary metabolites of three different-colored (white, pale green, and green) pak choi cultivars (Brassica rapa subsp. chinensis) using gas chromatography attached with time-of-flight mass spectrometry (GC-TOFMS) and high-performance liquid chromatography (HPLC). In total, 53 primary metabolites were identified and subjected to partial least-squares discriminant analysis. The result revealed a significant difference in the primary and secondary metabolites between the three pak choi cultivars. In addition, 49 hydrophilic metabolites were detected in different cultivars. Total phenolic and glucosinolate contents were highest in the pale green and green cultivars, respectively, whereas total carotenoid and chlorophyll contents were highest in the white cultivar. Superoxide dismutase activity, 2,2-diphenyl-1-picrylhydraz scavenging, and reducing power were slightly increased in the white, pale green, and green cultivars, respectively. In addition, a negative correlation between pigments and phenylpropanoids was discovered by metabolite correlation analysis. This approach will provide useful information for the development of strategies to enhance the biosynthesis of phenolics, glucosinolates, carotenoids, and chlorophyll, and to improve antioxidant activity in pak choi cultivars. In addition, this study supports the use of HPLC and GC-TOFMS-based metabolite profiling to explore differences in pak choi cultivars.

Từ khóa


Tài liệu tham khảo

Biala W, Jasinski M (2018) The phenylpropanoid case - It is transport that matters. Front Plant Sci 9:1610. https://doi.org/10.3389/fpls.2018.01610

Cevallos-Casals BA, Cisneros-Zevallos L (2010) Impact of germination on phenolic content and antioxidant activity of 13 edible seed species. Food Chem 119:1485–1490. https://doi.org/10.1016/j.foodchem.2009.09.030

Korkina L, Kostyuk V, De Luca C, Pastore S (2011) Plant phenylpropanoids as emerging anti-inflammatory agents. Mini-Rev Med Chem 11:823–835. https://doi.org/10.2174/138955711796575489

Panda P, Appalashetti M, Judeh ZMA (2011) Phenylpropanoid sucrose esters: Plant-derived natural products as potential leads for new therapeutics. Curr Med Chem 18:3234–3251. https://doi.org/10.2174/092986711796391589

Yang CS, Landau JM, Huang MT, Newmark HL (2001) Inhibition of carcinogenesis by dietary polyphenolic compounds. Annu Rev Nutr 21:381–406. https://doi.org/10.1146/annurev.nutr.21.1.381

Yao LH, Jiang YM, Shi J, Tomas-Barberan FA, Datta N, Singanusong R, Chen SS (2004) Flavonoids in food and their health benefits. Plant Food Hum Nutr 59:113–122. https://doi.org/10.1007/s11130-004-0049-7

Sathasivam R, Ki JS (2018) A review of the biological activities of microalgal carotenoids and their potential use in healthcare and cosmetic industries. Mar Drugs 16:26. https://doi.org/10.3390/md16010026

Sathasivam R, Ki JS (2019) Differential transcriptional responses of carotenoid biosynthesis genes in the marine green alga Tetraselmis suecica exposed to redox and non-redox active metals. Mol Biol Rep 46:1167–1179. https://doi.org/10.1007/s11033-018-04583-9

Park CH, Yeo HJ, Park SY, Kim JK, Park SU (2019) Comparative phytochemical analyses and metabolic profiling of different phenotypes of Chinese cabbage (Brassica rapa ssp. pekinensis). Foods 8:587. https://doi.org/10.3390/foods8110587

Sathasivam R, Radhakrishnan R, Kim JK, Park SU (2020) An update on biosynthesis and regulation of carotenoids in plants. S Afr J Bot. https://doi.org/10.1016/j.sajb.2020.05.015

Wang YT, Yang CH, Huang TY, Tai MH, Sie RH, Shaw JF (2019) Cytotoxic effects of chlorophyllides in ethanol crude extracts from plant leaves. Evid-Based Compl Alt 9494328:13. https://doi.org/10.1155/2019/9494328

Zhu B, Yang J, Zhu ZJ (2013) Variation in glucosinolates in pak choi cultivars and various organs at different stages of vegetative growth during the harvest period. J Zhejiang Univ-Sc B 14:309–317. https://doi.org/10.1631/jzus.B1200213

Barba FJ, Nikmaram N, Roohinejad S, Khelfa A, Zhu ZZ, Koubaa M (2016) Bioavailability of glucosinolates and their breakdown products: impact of processing. Front Nutr 3:24. https://doi.org/10.3389/fnut.2016.00024

Padilla G, Cartea ME, Velasco P, de Haro A, Ordas A (2007) Variation of glucosinolates in vegetable crops of Brassica rapa. Phytochemistry 68:536–545. https://doi.org/10.1016/j.phytochem.2006.11.017

Park CH, Yeo HJ, Baskar TB, Park YE, Park JS, Lee SY, Park SU (2019) In vitro antioxidant and antimicrobial properties of flower, leaf, and stem extracts of Korean mint. Antioxidants-Basel 8:75. https://doi.org/10.3390/antiox8030075

Mates JM, Perez-Gomez C, De Castro IN (1999) Antioxidant enzymes and human diseases. Clin Biochem 32:595–603. https://doi.org/10.1016/S0009-9120(99)00075-2

Garcia-Andrade M, Gonzalez-Laredo RF, Rocha-Guzman NE, Gallegos-Infante JA, Rosales-Castro M, Medina-Torres L (2013) Mesquite leaves (Prosopis laevigata), a natural resource with antioxidant capacity and cardioprotection potential. Ind Crop Prod 44:336–342. https://doi.org/10.1016/j.indcrop.2012.11.030

Giliberto L, Perrotta G, Pallara P, Weller JL, Fraser PD, Bramley PM, Fiore A, Tavazza M, Giuliano G (2005) Manipulation of the blue light photoreceptor cryptochrome 2 in tomato affects vegetative development, flowering time, and fruit antioxidant content. Plant Physiol 137:199–208. https://doi.org/10.1104/pp.104.051987

Schofield A, Paliyath G (2005) Modulation of carotenoid biosynthesis during tomato fruit ripening through phytochrome regulation of phytoene synthase activity. Plant Physiol Bioch 43:1052–1060. https://doi.org/10.1016/j.plaphy.2005.10.006

vonLintig J, Welsch R, Bonk M, Giuliano G, Batschauer A, Kleinig H (1997) Light-dependent regulation of carotenoid biosynthesis occurs at the level of phytoene synthase expression and is mediated by phytochrome in Sinapis alba and Arabidopsis thaliana seedlings. Plant J 12:625–634. https://doi.org/10.1046/j.1365-313X.1997.d01-16.x

Woitsch S, Romer S (2003) Expression of xanthophyll biosynthetic genes during light-dependent chloroplast differentiation. Plant Physiol 132:1508–1517. https://doi.org/10.1104/pp.102.019364

Zhang YJ, Chen GP, Dong TT, Pan Y, Zhao ZP, Tian SB, Hu ZL (2014) Anthocyanin accumulation and transcriptional regulation of anthocyanin biosynthesis in purple bok-choy (Brassica rapa var. chinensis). J Agr Food Chem 62:12366–12376. https://doi.org/10.1021/jf503453e

Bhandari SR, Jo JS, Lee JG (2015) Comparison of glucosinolate profiles in different tissues of nine Brassica crops. Molecules 20:15827–15841. https://doi.org/10.3390/molecules200915827

Harbaum B, Hubbermann EM, Wolff C, Herges R, Zhu Z, Schwarz K (2007) Identification of flavonoids and hydroxycinnamic acids in pak choi varieties (Brassica campestris L. ssp. chinensis var. communis) by HPLC-ESI-MSn and NMR and their quantification by HPLC-DAD. J Agric Food Chem 55:8251–8260. https://doi.org/10.1021/jf071314+

Harbaum B, Hubbermann EM, Zhu ZJ, Schwarz K (2008) Free and bound phenolic compounds in leaves of pak choi (Brassica campestris L. ssp chinensis var. communis) and Chinese leaf mustard (Brassica juncea Coss). Food Chem 110:838–846. https://doi.org/10.1016/j.foodchem.2008.02.069

Rochfort SJ, Imsic M, Jones R, Trenerry VC, Tomkins B (2006) Characterization of flavonol conjugates in immature leaves of pak choi [Brassica rapa L. Ssp chinensis L. (Hanelt.)] by HPLC-DAD and LC-MS/MS. J Agr Food Chem 54:4855–4860. https://doi.org/10.1021/jf060154j

Jeon J, Lim CJ, Kim JK, Park SU (2018) Comparative metabolic profiling of green and purple pakchoi (Brassica Rapa Subsp Chinensis). Molecules 23:613. https://doi.org/10.3390/molecules23071613

Park CH, Yeo HJ, Park YE, Baek SA, Kim JK, Park SU (2019) Transcriptome analysis and metabolic profiling of Lycoris radiata. Biology-Basel 8:63. https://doi.org/10.3390/biology8030063

Ha SH, Kim JK, Jeong YS, You MK, Lim SH, Kim JK (2019) Stepwise pathway engineering to the biosynthesis of zeaxanthin, astaxanthin and capsanthin in rice endosperm. Metab Eng 52:178–189. https://doi.org/10.1016/j.ymben.2018.11.012

Wellburn AR (1994) The spectral determination of chlorophyll-a and chlorophhyll-B, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313. https://doi.org/10.1016/S0176-1617(11)81192-2

Jeon J, Kim JK, Wu Q, Park SU (2018) Effects of cold stress on transcripts and metabolites in tartary buckwheat (Fagopyrum tataricum). Environ Exp Bot 155:488–496. https://doi.org/10.1016/j.envexpbot.2018.07.027

Kutmon M, van Iersel MP, Bohler A, Kelder T, Nunes N, Pico AR, Evelo CT (2015) PathVisio 3: An extendable pathway analysis toolbox. PLOS Comput Biol 11:e1004085. https://doi.org/10.1371/journal.pcbi.1004085

Steuer R, Kurths J, Fiehn O, Weckwerth W (2003) Interpreting correlations in metabolomic networks. Biochem Soc T 31:1476–1478. https://doi.org/10.1042/bst0311476

Cuong D, Ha TW, Park CH, Kim NS, Yeo HJ, Chun SW, Kim C, Park SU (2019) Effects of LED lights on expression of genes involved in phenylpropanoid biosynthesis and accumulation of phenylpropanoids in wheat sprout. Agronomy-Basel 9:307. https://doi.org/10.3390/agronomy9060307

Cuong D, Park SU, Park CH, Kim NS, Bong SJ, Lee SY (2019) Comparative analysis of glucosinolate production in hairy roots of green and red kale (Brassica oleracea var. acephala). Prep Biochem Biotech 49:775–782. https://doi.org/10.1080/10826068.2019.1615505

Tuan PA, Lee J, Park CH, Kim JK, Noh YH, Kim YB, Kim H, Park SU (2019) Carotenoid biosynthesis in oriental melon (Cucumis melo L. var. makuwa). Foods 8:77. https://doi.org/10.3390/foods8020077

Wiesner M, Zrenner R, Krumbein A, Glatt H, Schreiner M (2013) Genotypic variation of the glucosinolate profile in pak choi (Brassica rapa ssp chinensis). J Agr Food Chem 61:1943–1953. https://doi.org/10.1021/jf303970k

Chun JH, Kim NH, Seo MS, Jin M, Park SU, Arasu MV, Kim SJ, Al-Dhabi NA (2018) Molecular characterization of glucosinolates and carotenoid biosynthetic genes in Chinese cabbage (Brassica rapa L. ssp pekinensis). Saudi J Biol Sci 25:71–82. https://doi.org/10.1016/j.sjbs.2016.04.004

Kim JK, Chu SM, Kim SJ, Lee DJ, Lee SY, Lim SH, Ha SH, Kweon SJ, Cho HS (2010) Variation of glucosinolates in vegetable crops of Brassica rapa L. ssp pekinensis. Food Chem 119:423–428. https://doi.org/10.1016/j.foodchem.2009.08.051

Baek SA, Jung YH, Lim SH, Park SU, Kim JK (2016) Metabolic profiling in Chinese cabbage (Brassica rapa L. subsp pekinensis) cultivars reveals that glucosinolate content is correlated with carotenoid content. J Agr Food Chem 64:4426–4434. https://doi.org/10.1021/acs.jafc.6b01323

Watanabe M, Musumi K, Ayugase J (2011) Carotenoid pigment composition, polyphenol content, and antioxidant activities of extracts from orange-colored Chinese cabbage. LWT-Food Sci Technol 44:1971–1975. https://doi.org/10.1016/j.lwt.2011.04.010

Jiang N, Chung SO, Lee J, Ryu D, Lim YP, Park S, Lee C, Song J, Kim K, Park JT, An G (2013) Increase of phenolic compounds in new Chinese cabbage cultivar with red phenotype. Hortic Environ Biote 54:82–88. https://doi.org/10.1007/s13580-013-0136-5

Lee H, Oh IN, Kim J, Jung DH, Cuong NP, Kim Y, Lee J, Kwon O, Park SU, Lim Y, Kim B, Park JT (2018) Phenolic compound profiles and their seasonal variations in new red phenotype head-forming Chinese cabbages. LWT-Food Sci Technol 90:433–439. https://doi.org/10.1016/j.lwt.2017.12.056

Markom M, Hasan M, Daud WRW, Singh H, Jahim JM (2007) Extraction of hydrolysable tannins from Phyllanthus niruri Linn.: Effects of solvents and extraction methods. Sep Purif Technol 52:487–496. https://doi.org/10.1016/j.seppur.2006.06.003

Guo LP, Wang S, Zhang J, Yang G, Zhao MX, Ma WF, Zhang XB, Li X, Han BX, Chen N, Huang LQ (2013) Effects of ecological factors on secondary metabolites and inorganic elements of Scutellaria baicalensis and analysis of geoherblism. Sci China Life Sci 56:1047–1056. https://doi.org/10.1007/s11427-013-4562-5

Szakiel A, Paczkowski C, Henry M (2011) Influence of environmental abiotic factors on the content of saponins in plants. Phytochem Rev 10:471–491. https://doi.org/10.1007/s11101-010-9177-x

Park CH, Park SY, Lee SY, Kim JK, Park SU (2018) Analysis of metabolites in white flowers of Magnolia denudata Desr. and violet flowers of Magnolia liliiflora Desr. Molecules 23:1558. https://doi.org/10.3390/molecules23071558

Zhao SC, Park CH, Yang JL, Yeo HJ, Kim TJ, Kim JK, Park SU (2019) Molecular characterization of anthocyanin and betulinic acid biosynthesis in red and white mulberry fruits using high-throughput sequencing. Food Chem 279:364–372. https://doi.org/10.1016/j.foodchem.2018.11.101

Park CH, Morgan AMA, Park BB, Lee SY, Lee S, Kim JK, Park SU (2019) Metabolic analysis of four cultivars of Liriope platyphylla. Metabolites 9:59. https://doi.org/10.3390/metabo9030059

Kim JI, Dolan WL, Anderson NA, Chapple C (2015) Indole glucosinolate biosynthesis limits phenylpropanoid accumulation in Arabidopsis thaliana. Plant Cell 27:1529–1546. https://doi.org/10.1105/tpc.15.00127

Sakuta M, Hirano H, Kakegawa K, Suda J, Hirose M, Joy RW, Sugiyama M, Komamine A (1994) Regulatory mechanisms of biosynthesis of betacyanin and anthocyanin in relation to cell-division activity in suspension-cultures. Plant Cell Tiss Org 38:167–169. https://doi.org/10.1007/Bf00033874

Howles PA, Sewalt VJH, Paiva NL, Elkind Y, Bate NJ, Lamb C, Dixon RA (1996) Overexpression of L-phenylalanine ammonia-lyase in transgenic tobacco plants reveals control points for flux into phenylpropanoid biosynthesis. Plant Physiol 112:1617–1624. https://doi.org/10.1104/pp.112.4.1617

Blount JW, Korth KL, Masoud SA, Rasmussen S, Lamb C, Dixon RA (2000) Altering expression of cinnamic acid 4-hydroxylase in transgenic plants provides evidence for a feedback loop at the entry point into the phenylpropanoid pathway. Plant Physiol 122:107–116. https://doi.org/10.1104/pp.122.1.107

Muir SR, Collins GJ, Robinson S, Hughes S, Bovy A, De Vos CHR, van Tunen AJ, Verhoeyen ME (2001) Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols. Nat Biotechnol 19:470–474. https://doi.org/10.1038/88150

Briante R, Febbraio F, Nucci R (2003) Antioxidant properties of low molecular weight phenols present in the Mediterranean diet. J Agr Food Chem 51:6975–6981. https://doi.org/10.1021/jf034471r

Lee OH, Lee BY (2010) Antioxidant and antimicrobial activities of individual and combined phenolics in Olea europaea leaf extract. Bioresource Technol 101:3751–3754. https://doi.org/10.1016/j.biortech.2009.12.052

Lee KB, Kim YJ, Kim HJ, Choi J, Kim JK (2018) Phytochemical profiles of Brassicaceae vegetables and their multivariate characterization using chemometrics. Appl Biol Chem 61:131–144. https://doi.org/10.1007/s13765-017-0340-6

Park CH, Yeo HJ, Kim NS, Eun PY, Kim SJ, Arasu MV, Al-Dhabi NA, Park SY, Kim JK, Park SU (2017) Metabolic profiling of pale green and purple kohlrabi (Brassica oleracea var. gongylodes). Appl Biol Chem 60:249–257. https://doi.org/10.1007/s13765-017-0274-z

Park SY, Lim SH, Ha SH, Yeo Y, Park WT, Kwon DY, Park SU, Kim JK (2013) Metabolite profiling approach reveals the interface of primary and secondary metabolism in colored cauliflowers (Brassica oleracea L. ssp botrytis). J Agr Food Chem 61:6999–7007. https://doi.org/10.1021/jf401330e

Park S, Arasu MV, Jiang N, Choi SH, Lim YP, Park JT, Al-Dhabi NA, Kim SJ (2014) Metabolite profiling of phenolics, anthocyanins and flavonols in cabbage (Brassica oleracea var. capitata). Ind Crop Prod 60:8–14. https://doi.org/10.1016/j.indcrop.2014.05.037

Rosa E, Gomes MH (2002) Relationship between free amino acids and glucosinolates in primary and secondary inflorescences of 11 broccoli (Brassica oleracea L var italica) cultivars grown in early and late seasons. J Sci Food Agr 82:61–64. https://doi.org/10.1002/jsfa.999

Josefsson E (1970) Glucosinolate content and amino acid composition of rapeseed (Brassica napus) meal as affected by sulphur and nitrogen nutrition. J Sci Food Agr 21:98–103. https://doi.org/10.1002/jsfa.2740210211