Skip to main content
Log in

Haplotype analysis of the germacrene A synthase gene and association with cynaropicrin content and biological activities in Cynara cardunculus

  • Original Article
  • Published:
Molecular Genetics and Genomics Aims and scope Submit manuscript

Abstract

Cynara cardunculus: L. represents a natural source of terpenic compounds, with the predominant molecule being cynaropicrin. Cynaropicrin is gaining interest since it has been correlated to anti-hyperlipidaemia, antispasmodic and cytotoxicity activity against leukocyte cancer cells. The objective of this work was to screen a collection of C. cardunculus, from different origins, for new allelic variants in germacrene A synthase (GAS) gene involved in the cynaropicrin biosynthesis and correlate them with improved cynaropicrin content and biological activities. Using high-resolution melting, nine haplotypes were identified. The putative impact of the identified allelic variants in GAS protein was evaluated by bioinformatic tools and polymorphisms that putatively lead to protein conformational changes were described. Additionally, cynaropicrin and main pentacyclic triterpenes contents, and antithrombin, antimicrobial and antiproliferative activities were also determined in C. cardunculus leaf lipophilic-derived extracts. In this work we identified allelic variants with putative impact on GAS protein, which are significantly associated with cynaropicrin content and antiproliferative activity. The results obtained suggest that the identified polymorphisms should be explored as putative genetic markers correlated with biological properties in Cynara cardunculus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

a.a:

Aminoacid

CDS:

Coding sequence

Cc :

Cynara cardunculus L.

DMSO:

Dimethyl sulfoxide

EI-MS:

Electron impact mass spectra

GC–MS:

Gas chromatography–mass spectrometry

GAS:

Germacrene A synthase

HPLC–UV:

High-Performance Liquid Chromatography–Ultraviolet-Visible analysis

HRM:

High-resolution melting

MRSA:

Methicillin-Resistant Staphylococcus aureus

MBC:

Minimum bactericidal concentration

MIC:

Minimal inhibitory concentration

MAF:

Minor allele frequency

MH:

Mueller Hinton

PTM:

Post-translational modification

PROC GLM:

Procedure general linear model

SNP:

Single-nucleotide polymorphism

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

References

  • Atherinos AE, El-Sayed El-Kholy I, Soliman G (1962) Chemical investigation of Cynara scolymus L. Part I. The steroids of the receptacles and leaves. J Chem Soc 1700–1704

  • Bijak M, Saluk J, Ponczek MB, Nowak P (2013) Antithrombin effect of polyphenol-rich extracts from black chokeberry and grape seeds. Phyther Res 27:71–76

    Article  CAS  Google Scholar 

  • Blom N, Gammeltoft S, Brunak S (1999) Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. J Mol Biol 294:1351–1362

    Article  CAS  PubMed  Google Scholar 

  • Chadwick M, Trewin H, Gawthrop F, Wagstaff C (2013) Sesquiterpenoids lactones: benefits to plants and people. Int J Mol Sci 14:12780–12805

    Article  PubMed  PubMed Central  Google Scholar 

  • Chagné D, Gasic K, Crowhurst RN, Han Y, Bassett HC, Bowatte DR, Lawrence TJ, Rikkerink EHA, Gardiner SE, Korban SS (2008) Genomics development of a set of SNP markers present in expressed genes of the apple. Genomics 92:353–358

    Article  PubMed  Google Scholar 

  • Chen Y, Wilde HD (2011) Mutation scanning of peach floral genes. BMC Plant Biol 11:96

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen H, Xue Y, Huang N, Yao X, Sun Z (2006) MeMo: a web tool for prediction of protein methylation modifications. Nucleic Acids Res 34:249–253

    Article  Google Scholar 

  • Chen X, Qiu JD, Shi SP, Suo SB, Huang SY, Liang RP (2013) Incorporating key position and amino acid residue features to identify general and species-specific Ubiquitin conjugation sites. Bioinformatics 29:1614–1622

    Article  CAS  PubMed  Google Scholar 

  • Chistokhodo N, Nguyen C, Cal T, Kachirskaia I, Cunningham G, Miles DH (2002) Antithrombin activity of medicinal plants from central Florida. 81:pp 1–4

  • Cho JY, Kim AR, Jung JH, Chun T, Rhee MH, Yoo ES (2004) Cytotoxic and pro-apoptotic activities of cynaropicrin, a sesquiterpene lactone, on the viability of leukocyte cancer cell lines. Eur J Pharmacol 492:85–94

    Article  CAS  PubMed  Google Scholar 

  • Choi SZ, Choi SU, Lee KR (2005) Cytotoxic sesquiterpene lactones from Saussurea calcicola. Arch Pharm Res 28:1142–1146

    Article  CAS  PubMed  Google Scholar 

  • de Medeiros JM, Macedo M, Contancia JP, Nguyen C, Cunningham G, Miles DH (2000) Antithrombin activity of medicinal plants of the Azores. J Ethnopharmacol 72:157–165

    Article  PubMed  Google Scholar 

  • Dereeper A, Nicolas S, Le Cunff L, Bacilieri R, Doligez A, Peros J, Ruiz M, This P (2011) SNiPlay : a web-based tool for detection, management and analysis of SNPs. Application to grapevine diversity projects. BMC Bioinformatics 12:134

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Djeussi DE, Noumedem JA, Seukep J a, Fankam AG, Voukeng IK, Tankeo SB, Nkuete AH, Kuete V (2013) Antibacterial activities of selected edible plants extracts against multidrug-resistant Gram-negative bacteria. BMC Complement Altern Med 13:164

    Article  PubMed  PubMed Central  Google Scholar 

  • El Solh A a., Alhajhusain A (2009) Update on the treatment of Pseudomonas aeruginosa pneumonia. J Antimicrob Chemother 64:229–238

    Article  PubMed  Google Scholar 

  • Eljounaidi K, Cankar K, Comino C, Moglia A, Hehn A, Bourgaud F, Bouwmeester H, Menin B, Lanteri S, Beekwilder J (2014) Cytochrome P450s from Cynara cardunculus L. CYP71AV9 and CYP71BL5, catalyze distinct hydroxylations in the sesquiterpene lactone biosynthetic pathway. Plant Sci 223:59–68

    Article  CAS  PubMed  Google Scholar 

  • Eljounaidi K, Comino C, Moglia a., Cankar K, Genre a., Hehn a., Bourgaud F, Beekwilder J, Lanteri S (2015) Accumulation of cynaropicrin in globe artichoke and localization of enzymes involved in its biosynthesis. Plant Sci 239:128–136

    Article  CAS  PubMed  Google Scholar 

  • Eloff JN (1998) A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria. Planta Med 64:711–713

    Article  CAS  PubMed  Google Scholar 

  • Elsebai MF, Koutsoudakis G, Saludes V, Pérez-Vilaró G, Turpeinen A, Mattila S, Pirttilä AM, Fontaine-Vive F, Mehiri M, Meyerhans A, Diez J (2016) Pan-genotypic hepatitis C virus inhibition by natural products derived from the wild Egyptian artichoke. J Virol 90:1918–1930

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Emendörfer F, Emendörfer F, Bellato F, Noldin VF, Cechinel-Filho V, Yunes RA, Delle Monache F, Cardozo AM (2005) Antispasmodic activity of fractions and cynaropicrin from Cynara scolymus on guinea-pig ileum. Biol Pharm Bull 28:902–904

    Article  PubMed  Google Scholar 

  • Falleh H, Ksouri R, Chaieb K, Karray-Bouraoui N, Trabelsi N, Boulaaba M, Abdelly C (2008) Phenolic composition of Cynara cardunculus L. organs, and their biological activities. C R Biol 331:372–379

    Article  CAS  PubMed  Google Scholar 

  • Farooqui A, Khan A, Borghetto I, Kazmi SU, Rubino S, Paglietti B (2015) Synergistic antimicrobial activity of Camellia sinensis and Juglans regia against multidrug-resistant bacteria. PLoS One 10:e0118431

    Article  PubMed  PubMed Central  Google Scholar 

  • Fernández J, Curt MD, Aguado PL (2006) Industrial applications of Cynara cardunculus L. for energy and other uses. Ind Crops Prod 24:222–229

    Article  Google Scholar 

  • Ferro AM, Ramos P, Guerreiro O, Jerónimo E, Pires I, Capel C, Capel J, Lozano R, Duarte MF, Oliveira MM, Gonçalves S (2017) Impact of novel SNPs identified in Cynara cardunculus genes on functionality of proteins regulating phenylpropanoid pathway and their association with biological activities. BMC Genom 18:183

    Article  Google Scholar 

  • Ganopoulos I, Argiriou A, Tsaftaris A (2011) Microsatellite high resolution melting (SSR-HRM) analysis for authenticity testing of protected designation of origin (PDO) sweet cherry products. Food Control 22:532–541

    Article  CAS  Google Scholar 

  • Gominho J, Fernandez J, Pereira H (2001) Cynara cardunculus L.—a new fibre crop for pulp and paper production. Ind Crops Prod 13:1–10

    Article  Google Scholar 

  • Goudarzi M, Fazeli M, Azad M, Seyedjavadi SS, Mousavi R (2015) Aloe vera gel: effective therapeutic agent against multidrug-resistant Pseudomonas aeruginosa isolates recovered from burn wound infections

  • Han Y, Khu D-M, Monteros MJ (2012) High-resolution melting analysis for SNP genotyping and mapping in tetraploid alfalfa (Medicago sativa L.). Mol Breed 29:489–501

    Article  PubMed  Google Scholar 

  • Japoni A, Farshad S, Alborzi a (2009) Pseudomonas aeruginosa: burn infection, treatment and antibacterial resistance. Iran Red Crescent Med J 11:244–253

    Google Scholar 

  • Jeong H-J, Jo YD, Park S-W, Kang B-C (2010) Identification of Capsicum species using SNP markers based on high resolution melting analysis. Genome 53:1029–1040

    Article  CAS  PubMed  Google Scholar 

  • Johannes F, Porcher E, Teixeira FK, Saliba-Colombani V, Simon M, Agier N, Bulski A, Albuisson J, Heredia F, Audigier P, Bouchez D, Dillmann C, Guerche P, Hospital F, Colot V, Visscher PM (2009) Assessing the impact of transgenerational epigenetic variation on complex traits. PLoS Genet 5(6):e1000530

    Article  PubMed  PubMed Central  Google Scholar 

  • Jung M-Y, Chung J-Y, Lee H-Y, Park J, Lee D-Y, Yang J-M (2015) Antibiotic susceptibility of Staphylococcus aureus in atopic dermatitis: current prevalence of methicillin-resistant Staphylococcus aureus in Korea and treatment strategies. Ann Dermatol 27:398–403

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kammoun M, Koubaa I, Ben Ali Y, Jarraya R, Gargouri Y, Damak M, Bezzine S (2010) Inhibition of pro-inflammatory secreted phospholipase A2 by extracts from Cynara cardunculus L. Appl Biochem Biotechnol 162:662–670

    Article  CAS  PubMed  Google Scholar 

  • Kotiranta A, Lounatmaa K, Haapasalo M (2000) Epidemiology and pathogenesis of Bacillus cereus infections. Microbes Infect 2:189–198

    Article  CAS  PubMed  Google Scholar 

  • Lehmensiek A, Sutherland MW, Mcnamara RB (2008) The use of high resolution melting (HRM) to map single nucleotide polymorphism markers linked to a covered smut resistance gene in barley. 721–728

  • Lhinhatrakool T, Sutthivaiyakit S (2006) 19-Nor- and 18,20-epoxy-cardenolides from the leaves of Calotropis gigantea. J Nat Prod 69:1249–1251

    Article  CAS  PubMed  Google Scholar 

  • Mackay JF, Wright CD, Bonfiglioli RG (2008) A new approach to varietal identification in plants by microsatellite high resolution melting analysis: application to the verification of grapevine and olive cultivars. Plant Methods 4:8

    Article  PubMed  PubMed Central  Google Scholar 

  • Menin B, Comino C, Portis E, Moglia A, Cankar K, Bouwmeester HJ, Lanteri S, Beekwilder J (2012) Genetic mapping and characterization of the globe artichoke (+)-germacrene A synthase gene, encoding the first dedicated enzyme for biosynthesis of the bitter sesquiterpene lactone cynaropicrin. Plant Sci 190:1–8

    Article  CAS  PubMed  Google Scholar 

  • Miccadei S, Di Venere D, Cardinali A, Romano F, Durazzo A, Foddai MS, Fraioli R, Mobarhan S, Maiani G (2008) Antioxidative and apoptotic properties of polyphenolic extracts from edible part of artichoke (Cynara scolymus L.) on cultured rat hepatocytes and on human hepatoma cells. Nutr Cancer 60:276–283

    Article  CAS  PubMed  Google Scholar 

  • Mileo AM, Di Venere D, Linsalata V, Fraioli R, Miccadei S (2012) Artichoke polyphenols induce apoptosis and decrease the invasive potential of the human breast cancer cell line MDA-MB231. J Cell Physiol 227:3301–3309

    Article  CAS  PubMed  Google Scholar 

  • Mileo AM, Di Venere D, Abbruzzese C, Miccadei S (2015) Long term exposure to polyphenols of artichoke (Cynara scolymus L.) exerts induction of senescence driven growth arrest in the MDA-MB231 human breast cancer cell line. Oxid Med Cell Longev 2015:1–11

  • Mondini L, Nachit M, Porceddu E, Pagnotta MA (2012) Identification of SNP mutations in DREB1, HKT1, and WRKY1 genes involved in drought and salt stress tolerance in durum wheat (Triticum turgidum L. var durum). OMICS 16:178–187

    Article  CAS  PubMed  Google Scholar 

  • Mossi A, Echeverrigaray S (1999) Identification and characterization of antimicrobial components in leaf extracts of globe artichoke (Cynara scolymus L.). Acta Hortic 501:111–114

    Article  Google Scholar 

  • Mota I, Rodrigues Pinto PC, Novo C, Sousa G, Guerreiro O, Guerra ÂR, Duarte MF, Rodrigues AE (2012) Extraction of polyphenolic compounds from Eucalyptus globulus bark: process optimization and screening for biological activity. Ind Eng Chem Res 51:6991–7000

    Article  CAS  Google Scholar 

  • Muhammad I, Takamatsu S, Mossa JS, El-Feraly FS, Walker LA, Clark AM (2003) Cytotoxic sesquiterpene lactones from Centaurothamnus maximus and Vicoa pentanema. Phyther Res 17:168–173

    Article  CAS  Google Scholar 

  • Muleo R, Colao MC, Miano D, Cirilli M, Intrieri MC, Baldoni L, Rugini E (2009) Mutation scanning and genotyping by high-resolution DNA melting analysis in olive germplasm. Genome 52:252–260

    Article  CAS  PubMed  Google Scholar 

  • Noldin VF, Cechinel Filho V, Monache FD, Benassi JC, Christmann IL, Pedrosa RC, Yunes RA (2003) Composição química e atividades biológicas das folhas de Cynara scolymus L. (alcachofra) cultivada no Brasil. Química Nov 26:331–334

    Article  CAS  Google Scholar 

  • Portis E, Barchi L, Acquadro A, Macua J, Lanteri S (2005) Genetic diversity assessment in cultivated cardoon by AFLP (amplified fragment length polymorphism) and microsatellite markers. Plant Breed 124:299–304

    Article  CAS  Google Scholar 

  • Raccuia S a., Melilli MG (2007) Biomass and grain oil yields in Cynara cardunculus L. genotypes grown in a Mediterranean environment. F Crop Res 101:187–197

    Article  Google Scholar 

  • Radivojac P, Vacic V, Haynes C, Cocklin RR, Mohan A, Heyen JW, Goebl MG, Iakoucheva LM (2010) Identification, analysis, and prediction of protein ubiquitination sites. Proteins 78:365–380

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ramos P a B, Guerra ÂR, Guerreiro O, Freire CSR, Silva AMS, Duarte MF, Silvestre AJD (2013) Lipophilic extracts of Cynara cardunculus L. var. altilis (DC): a source of valuable bioactive terpenic compounds. J Agric Food Chem 61:8420–8429

    Article  CAS  PubMed  Google Scholar 

  • Ramos PAB, Santos SAO, Guerra ÂR, Guerreiro O, Freire CSR, Rocha SM, Duarte MF, Silvestre AJD (2014) Phenolic composition and antioxidant activity of different morphological parts of Cynara cardunculus L. var. altilis (DC) Patrícia A. B. Ramos. Ind Crop Prod 61:1–47

    Article  Google Scholar 

  • Ramos P, Guerra Â, Guerreiro O, Santos S, Oliveira H, Freire C, Silvestre A, Duarte M (2016) Antiproliferative effects of Cynara cardunculus L. var. altilis (DC) lipophilic extracts. Int J Mol Sci 18:63

    Article  PubMed Central  Google Scholar 

  • Schwienhorst A (2006) Direct thrombin inhibitors—a survey of recent developments. Cell Mol Life Sci C 63:2773–2791

    Article  CAS  Google Scholar 

  • Shimoda H, Ninomiya K, Nishida N, Yoshino T, Morikawa T, Matsuda H, Yoshikawa M (2003) Anti-hyperlipidemic sesquiterpenes and new sesquiterpene glycosides from the leaves of artichoke (Cynara scolymus L.): structure requirement and mode of action. Bioorg Med Chem Lett 13:223–228

    Article  CAS  PubMed  Google Scholar 

  • Sonnante G, Gatto A, Morgese A, Montemurro F, Sarli G, Blanco E, Pignone D (2011) Genetic map of artichoke × wild cardoon: toward a consensus map for Cynara cardunculus. Theor Appl Genet 123:1215–1229

    Article  PubMed  Google Scholar 

  • Suchý M, Herout V, Šorm F (1960) On terpenes. CVI: on hydrogenation products of cynaropicrin, the bitter principle of artichoke (Cynara scolymus L.). Collect Czech Chem Commun 25:507–514

    Article  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tan YY, Fu HW, Zhao HJ, Lu S, Fu JJ, Li YF, Cui HR, Shu QY (2013) Functional molecular markers and high-resolution melting curve analysis of low phytic acid mutations for marker-assisted selection in rice. Mol Breed 31:517–528

    Article  CAS  Google Scholar 

  • Veríssimo P, Esteves C, Faro C, Pires E (1995) The vegetable rennet of Cynara cardunculus L. contains two proteinases with chymosin and pepsin-like specificities. Biotechnol Lett 17:621–626

    Article  Google Scholar 

  • Verspohl EJ, Ploch M, Windeck T, Klaes M, Schmidt T, Bauer K (2008) Effect of two artichoke extracts (36_U and 36_EB) on rat ileum (with respect to bowel syndrome) and the peristaltic threshold. Phytomedicine 15:1002–1009

    Article  CAS  PubMed  Google Scholar 

  • Wiegand I, Hilpert K, Hancock REW (2008) Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc 3:163–175

    Article  CAS  PubMed  Google Scholar 

  • Yasukawa K, Matsubara H, Sano Y (2010) Inhibitory effect of the flowers of artichoke (Cynara cardunculus) on TPA-induced inflammation and tumor promotion in two-stage carcinogenesis in mouse skin. J Nat Med 64:388–391

    Article  CAS  PubMed  Google Scholar 

  • Youngson NA, Whitelaw E (2008) Transgenerational epigenetic effects. Annu Rev Genomics Hum Genet 9(1):233–257

    Article  CAS  PubMed  Google Scholar 

  • Yuan YV, Carrington MF, Walsh NA (2005) Extracts from dulse (Palmaria palmata) are effective antioxidants and inhibitors of cell proliferation in vitro. Food Chem Toxicol 43:1073–1081

    Article  CAS  PubMed  Google Scholar 

  • Zhao J (2007) Nutraceuticals, nutritional therapy, phytonutrients, and phytotherapy for improvement of human health: a perspective on plant biotechnology application. Recent Pat Biotechnol 1:75–97

    Article  CAS  PubMed  Google Scholar 

  • Zuo GY, Wang GC, Zhao YB, Xu GL, Hao XY, Han J, Zhao Q (2008) Screening of Chinese medicinal plants for inhibition against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). J Ethnopharmacol 120:287–290

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Plant seeds were kindly provided by Botanical Garden of University of Tuscia (Italy), Botanical Garden of University of Agronomic Sciences of Gembloux (Belgium), Botanical Garden of University of Szeged (Hungary), Botanical Garden of University of Napoli (Italy), Experimental Centre of School of Agriculture of Beja (Portugal) and Botanical Garden of Sapienza University of Rome (Italy). We are also grateful to the Horticultural Centre of School of Agriculture (ESA) of Beja, for space to grow Cc plants. Pseudomonas aeruginosa POAI was kindly donated by Professor Arsénio Fialho from Instituto Superior Técnico; Methicillin-resistant Staphylococcus aureus (clinical isolate from wound exudate) and Bacillus cereus were both kindly donated by Professor Manuela Pintado from Universidade Católica Portuguesa. We also thank Conceição Fernandes for HPLC support, Inês Pires for TASSEL software support and Anabel Usié Chimenos and Marcos Ramos for bioinformatic support. Fundação para a Ciência e a Tecnologia (FCT) is gratefully acknowledged for the PhD fellowships, refs. SFRH/BD/68847/2010; SFRH/BD/70845/2010, SFRH/BD/84406/2012, SFRH/BD/98635/2013 and SFRH/BD/110969/2015 awarded to AF, PR, OG, AG and TB, respectively. SG was supported by Programa Ciência 2008, also funded by POPH (QREN). This project was funded by European Regional Development Fund (FEDER), ValBioTecCynara (ALT20-03-0145-FEDER-000038)—Economic valorization of Cardoon (Cynara cardunculus): study of natural variability and biotechnological applications. Thanks are also due to GREEN-it “Bioresources for sustainability” (UID/Multi/04551/2013). The authors also thank Junta de Andalucía (grant P12-AGR-1482) and Campus de Excelencia Internacional Agroalimentario (CeiA3) for supporting this research collaboration.

Author information

Authors and Affiliations

Authors

Contributions

AMF performed the experimental work and carried out the bioinformatic data analysis. PR contributed for GC-MS and GC-FID planning. AG contributed for antithrombin and antiproliferative activities and PP contributed for the antimicrobial activity. TB helped in the preparation of leaf lipophilic-derived extracts and cynaropicrin quantification by HPLC. OG and EJ contributed for statistical analysis. CC, JC and FJY-L contributed for HRM planning and execution. MFD, MMO, SG and RL contributed for the research planning, funding, discussions and paper writing. All authors read and approved the final manuscript

Corresponding author

Correspondence to Sónia Gonçalves.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal studies

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

Informed consent

Informed consent was obtained from all individual participants included in the study.

Additional information

Communicated by S. Hohmann.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Cynara cardunculus accessions used in this study (PDF 57 KB)

438_2017_1388_MOESM2_ESM.pdf

GAS gene structure and primer pairs (triangle arrows) designed for HRM analysis. (*)SNPs position; (#) a.a. alteration. The introns are indicated in gray (PDF 98 KB)

Primer pairs used in HRM analysis (PDF 86 KB)

438_2017_1388_MOESM4_ESM.pdf

Haplotypes identified for GAS gene. Individuals A1, A2, A4, B3, C1, D2, G3, J3, K3, U1, Y1, AB1 and AC2 were selected for phenotypic trait analysis and are indicated in gray. Some of these individuals present two haplotypes for GAS gene and are denominated X_1 and X_2. For instance, the denomination A2_1 and A2_2 correspond to the two haplotypes of the same individual A2, which was selected for phenotypic analysis (results presented in table 3) (PDF 113 KB)

SNPs identified on GAS gene (PDF 53 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ferro, A.M., Ramos, P., Guerra, Â. et al. Haplotype analysis of the germacrene A synthase gene and association with cynaropicrin content and biological activities in Cynara cardunculus . Mol Genet Genomics 293, 417–433 (2018). https://doi.org/10.1007/s00438-017-1388-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00438-017-1388-z

Keywords

Navigation