Abstract
The species Piper hispidinervum, Piper aduncum, and Piper affinis hispidinervum have essential oils with high levels of safrole, dillapiole, and sarisan, respectively. Safrole is important for pharmaceutical and chemical industries, while dillapiole and sarisan are promising compounds to control insects and fungi. These species are very similar morphologically and their taxonomy is controversial. Divergent hypotheses consider P. aduncum and P. hispidinervum either as a single species or as distinct taxa, while P. affinis hispidinervum is inferred to be a natural hybrid or a chemotype of P. hispidinervum. Delimiting the taxonomic boundaries would be helpful for germplasm conservation and breeding programs. This study aimed to undertake a detailed analysis of P. aduncum, P. hispidinervum, and P. affinis hispidinervum karyotype and rDNA sites. Genomic in situ hybridization (GISH) was used to establish genomic homology among species and to test the natural hybridization hypothesis for origin of P. affinis hispidinervum. Karyotype traits were similar for all three species: 2n = 26 small chromosomes, predominantly metacentric. All three species exhibited CMA+ bands on the secondary constriction of chromosome pair 4. A size-heteromorphic 35S rDNA site was co-localized with the CMA+ band. A 5S rDNA site was located in the proximal region of chromosome pair 7. The patterns of genomic hybridization revealed that the repetitive DNA fraction of the species is highly similar in terms of proportion of genome, sequence type, and distribution. Our findings did not allow us to differentiate the three species and point to the importance of deeper genomic studies to elucidate the taxonomic controversy.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Not applicable
Code availability
Not applicable
References
Altınordu F, Peruzzi L, Yu Y, He X (2016) A tool for the analysis of chromosomes : karyotype. Taxon 10(12705/653):9
Báez M, Souza G, Guerra M (2020) Does the chromosomal position of 35S rDNA sites influence their transcription? A survey on Nothoscordum species (Amaryllidaceae). Genet Mol Biol 43:1. https://doi.org/10.1590/1678-4685-GMB-2018-0194
Bai GVS, Subramanian D (1985) Cytotaxonomical studies of South Indian Piperaceae. Citologia 50:583–592. https://doi.org/10.1508/cytologia.50.583
Belyayev A, Raskina O, Nevo E (2001) Chromosomal distribution of reverse transcriptase-containing retroelements in two Triticeae species. Chromosome Res 9:129–136. https://doi.org/10.1023/A:1009231019833
Berjano R, Roa F, Talavera S, Guerra M (2009) Cytotaxonomy of diploid and polyploid Aristolochia (Aristolochiaceae) species based on the distribution of CMA/DAPI bands and 5S and 45S rDNA sites. Plant Syst Evol 280:219–227. https://doi.org/10.1007/s00606-009-0184-6
Bhowmick BK, Jha S (2019) Differences in karyotype and fluorochrome banding patterns among variations of trichosanthes cucumerina with different fruit size. Cytologia 84:237–245. https://doi.org/10.1508/cytologia.84.237
Brasileiro-Vidal AC, dos Santos-Serejo JA, dos S Soares Filho W, Guerra M (2007) A simple chromosomal marker can reliably distinguishes Poncirus from Citrus species. Genetica 129:273–279. https://doi.org/10.1007/s10709-006-0007-4
Chase MW, Christenhusz MJM, Fay MF et al (2016) An update of the angiosperm phylogeny group classification for the orders and families of flowering plants: APG IV. Bot J Linn Soc 181:1–20. https://doi.org/10.1111/boj.12385
Dantas LG, Guerra M (2010) Chromatin differentiation between Theobroma cacao L. and T. grandiflorum Schum. Genet Mol Biol 33:94–98. https://doi.org/10.1590/S1415-47572009005000103
Dasgupta A, Datta PC (1976) Cytotaxonomy Piperaceae. Cytologia 41:697–706. https://doi.org/10.1508/cytologia.41.697
de Figueiredo RA, Sazima M (2000) Pollination biology of Piperaceae species in Southeastern Brazil. Ann Bot 85:455–460. https://doi.org/10.1006/anbo.1999.1087
D’Hont A (2005) Unraveling the genome structure of polyploids using FISH and GISH; examples of sugarcane and banana. Cytogenet Genome Res 109:27–33. https://doi.org/10.1159/000082378
Dolezel J, Bartos J, Voglmayr H, Greilhuber J (2003) Nuclear DNA content and genome size of trout and human. Cytometry A 51:127–8. https://doi.org/10.1002/cyto.a.10013
Dong F, McGrath JM, Helgeson JP, Jiang J (2001) The genetic identity of alien chromosomes in potato breeding lines revealed by sequential GISH and FISH analyses using chromosome-specific cytogenetic DNA markers. Genome 44:729–734. https://doi.org/10.1139/gen-44-4-729
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15
Fazolin M, Estrela JLV, Catani V et al (2007) Propiedade inseticida dos óleo essenciais Piper hispidinervum C DC; Piper aduncum L. e Tanaecium nocturnum, (Bur, Rodr.) Shum, K. Sobre Tenebrio molitor L. 1758. Ciência e Agrotecnologia, 31(1):113–120
Fazolin M, Lima J, Estrela V, Catani V (2011) Avaliação toxicológica do óleo essencial de Piper affinis hispidinervum para insetos pragas das culturas do milho e feijão. In: In: CONGRESSO BRASILEIRO DE DEFENSIVOS AGRÍCOLAS NATURAIS, 5., 2011, Jaguariúna. [Anais...]. Jaguariúna: Embrapa Meio Ambiente, 2011. 1 CD-ROM.
Ferreira DF (2019) SISVAR: a computer analysis system to fixed effects split plot type designs. Rev Bras Biom 37:529–535. https://doi.org/10.28951/rbb.v37i4.450
Foysol AM, Bonna IJ, Alam SS, Sultana SS (2019) Karyotype diversity in three medicinal piper species in Bangladesh. Cytologia 84:63–67. https://doi.org/10.1508/cytologia.84.63
Gaiero P, Mazzella C, Vilaró F et al (2017) Pairing analysis and in situ hybridisation reveal autopolyploid-like behaviour in Solanum commersonii × S. tuberosum (potato) interspecific hybrids. Euphytica 213:137. https://doi.org/10.1007/s10681-017-1922-4
Galbraith DW, Lambert GM, Macas J, Dolezel J (1997) Analysis of nuclear DNA content and ploidy in higher plants. Curr Protoc Cytom 2:7.6.1-7.6.22. https://doi.org/10.1002/0471142956.cy0706s02
Gerbault-Seureau M, Cacheux L, Dutrillaux B (2018) The relationship between the (In-)stability of NORs and their chromosomal location: the example of Cercopithecidae and a short review of other primates. Cytogenet Genome Res 153:138–146. https://doi.org/10.1159/000486441
Guimarães EF, Medeiros EVSS, Queiroz GA (2020) Piper. In: Flora do Brasil. http://floradobrasil.jbrj.gov.br/reflora/floradobrasil/FB12735. Accessed 25 Sep 2021
Jaramillo MA, Marquis R (2004) Future research in piper biology. In: Piper: a model genus for studies of phytochemistry, ecology, and evolution (pp 199–203). Springer, Boston, MA
Jenkins G, Mur L, Bablak P, et al (2004) Prospects for functional genomics in a new model grass. In: Plant Functional Genomics. Taylor & Francis
Jeridi M, Bakry F, Escoute J et al (2011) Homoeologous chromosome pairing between the A and B genomes of Musa spp. revealed by genomic in situ hybridization. Ann Bot 108:975–981. https://doi.org/10.1093/aob/mcr207
Jha TB (2019) Karyotype analysis from aerial roots of Piper nigrum based on giemsa and fluorochrome banding. Cytologia 84:313–317. https://doi.org/10.1508/cytologia.84.313
Jiang J, Gill BS (2006) Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research. Genome 49:1057–1068. https://doi.org/10.1139/G06-076
Jose J, Sharma AK (1985) Structure and behaviour of chromosomes in Piper and Peperomia (family Piperaceae). Cytologia 50:301–310. https://doi.org/10.1508/cytologia.50.301
Jose J, JohnE T, Mathew L (1992) Chromosome complement Peperomia analysis in five species of Ruiz and Pav. Cytologia 57:227–229. https://doi.org/10.1508/cytologia.57.227
Joseph A, Joseph R, Jose J (1998) Karyomorphometrical analysis in Piper species using image analysis system. Cytobios. https://doi.org/10.1508/cytologia.64.313
Kemprai P, Mahanta BP, Sut D et al (2020) Review on safrole: identity shift of the ‘candy shop’ aroma to a carcinogen and deforester. Flavour Fragr J 35:5–23. https://doi.org/10.1002/FFJ.3521
VAN Laere K, Khrustaleva L, VAN Huylenbroeck J, VAN Bockstaele E (2010) Application of GISH to characterize woody ornamental hybrids with small genomes and chromosomes. Plant Breeding 447:442–447. https://doi.org/10.1111/j.1439-0523.2009.01692.x
Lapitan NLV (1992) Organization and evolution of higher plant nuclear genomes. Genome 35:171–181. https://doi.org/10.1139/g92-028
Levan A, Fredga K, Sandberg AA (1964) Nomeclature for Centromeric Position on Chromosomes. Hereditas 52:201–220. https://doi.org/10.1111/j.1601-5223.1964.tb01953.x
Li B, Zhang M, Ge S, Lu R (2001) Identification of genome constitution of Oryza malampuzhaensis, O. minuta, and O. punctata by multicolor genomic in situ hybridization. Theor Appl Genet 103:204–211. https://doi.org/10.1007/s001220100563
Lim K-B, de Jong H, Yang T-J, et al (2005) Characterization of rDNAs and tandem repeats in the heterochromatin of Brassica rapa. Mol Cells 19(3):436–444
Lima LM (2015) Safrole and the versatility of a natural biophore. Revista Virtual De Quimica 7:495–538. https://doi.org/10.5935/1984-6835.20150023
Maluszynska J, Hasterok R (2005) Identification of individual chromosomes and parental genomes in Brassica juncea using GISH and FISH. Cytogenet Genome Res 314:310–314. https://doi.org/10.1159/000082414
Mathew PJ, Mathew PM, Pushpangadan P (1999) Cytology and its bearing on the systematics and phylogeny of the Piperaceae. Cytologia 64:301–307. https://doi.org/10.1508/cytologia.64.301
Maugini E (1950) Ricerche cito-embriologiche sul genere piper. Caryologia 3:221–233. https://doi.org/10.1080/00087114.1950.10797159
Nunes JD, Torres GA, Davide LC, Salgado CC (2007) Citogenética de Piper hispidinervum e Piper aduncum. Pesquisa Agropecuaria Brasileira 42:1049–1052. https://doi.org/10.1590/S0100-204X2007000700019
dos Santos de Oliveira JA, Rodrigues DW (2021) ÓLEOS ESSENCIAIS DE PIPER L. (PIPERACEAE) E SUA APLICAÇÃO BIOTECNOLÓGICA NA AGRICULTURA: UMA REVISÃO DA LITERATURA. Arquivos Do Mudi 25:100–110. https://doi.org/10.4025/arqmudi.v25i2.60107
Oltramari AC, Wood KV, Bonham C et al (2004) Safrole analysis by GC-MS of prototrophic (Ocotea odorifera (Vell.) Rohwer) cell cultures. Plant Cell Tissue Organ Cult 78:231–235. https://doi.org/10.1023/B:TICU.0000025657.42171.71
Paszko B (2006) A critical review and a new proposal of karyotype asymmetry indices. Plant Syst Evol 258:39–48. https://doi.org/10.1007/s00606-005-0389-2
Pedrosa-Harand A, de Almeida CCS, Mosiolek M et al (2006) Extensive ribosomal DNA amplification during Andean common bean (Phaseolus vulgaris L.) evolution. Theor Appl Genet 112:924–933. https://doi.org/10.1007/s00122-005-0196-8
Pimentel FA et al. (1998) Recomendações para a produção de mudas de pimenta longa no Estado do Acre. Rio Branco
Piperidis G, Piperidis N, D’Hont A (2010) Molecular cytogenetic investigation of chromosome composition and transmission in sugarcane. Mol Genet Genomics 284:65–73. https://doi.org/10.1007/s00438-010-0546-3
Ribeiro T, Nascimento J, Santos A et al (2021) Origin and evolution of highly polymorphic rDNA sites in Alstroemeria longistaminea (Alstroemeriaceae) and related species. Genome 64:833–845. https://doi.org/10.1139/gen-2020-0159
Roa F, Guerra M (2012) Distribution of 45S rDNA sites in chromosomes of plants : structural and evolutionary implications. BMC Evol Biol 12:225. https://doi.org/10.1186/1471-2148-12-225
Roa F, Guerra M (2015) Non-random distribution of 5S rDNA sites and its association with 45S rDNA in plant chromosomes. Cytogenet Genome Res 146:243–249. https://doi.org/10.1159/000440930
Rosato M, Álvarez I, Nieto Feliner G, Rosselló JA (2017) High and uneven levels of 45S rDNA site-number variation across wild populations of a diploid plant genus (Anacyclus, Asteraceae). PLoS ONE 12:e0187131. https://doi.org/10.1371/journal.pone.0187131
Samuel R, Smith JB, Bennett MD (1986) Canadian Journal of Genetics and Cytology 28(6):1041–1043. https://doi.org/10.1139/g86-145
Samuel R, Morawetz W (1989) Chromosomal evolution within Piperaceae. Plant Systematics and Evolution 1989 166:1 166:105–117. https://doi.org/10.1007/BF00937879
Sastri DC, Hilu K, Appels R et al (1992) An overview of evolution in plant 5S DNA. Plant Syst Evol 183:169–181
Sharma AK, Bhattacharyya NK (1959) Chromosome studies on two genera of the family piperaceae. Genetica 29:256–289. https://doi.org/10.1007/BF01535714
Silva ACPR da, Oliveira MN de (2000) Caracterização botânica e química de três espécies do gênero Piper no Acre
Silva GS, Souza MM, de Melo CAF et al (2018) Identification and characterization of karyotype in Passiflora hybrids using FISH and GISH. BMC Genet 19:1–11. https://doi.org/10.1186/s12863-018-0612-0
Silva MHL (1993) Tecnologia de cultivo e produção racional de pimenta longa, Piper hispidinervium C. DC. Universidade Federal do Rio de Janeiro
Soltis DE, Soltis PS, Bennett MD, Leitch IJ (2003) Evolution of genome size in the angiosperms. Am J Bot 90:1596–1603. https://doi.org/10.3732/ajb.90.11.1596
Taher M, Amri MS, Susanti D, et al (2020) Medicinal Uses, Phytochemistry and Pharmacological Properties of Piper aduncum L. Sains Malaysiana 49:1829–1851. https://doi.org/10.17576/jsm-2020-4908-07
Vaio M, Mazzella C, Guerra M, Speranza P (2019) Effects of the diploidisation process upon the 5S and 35S rDNA sequences in the allopolyploid species of the Dilatata group of Paspalum (Poaceae, Paniceae). Aust J Bot 67:521–530. https://doi.org/10.1071/BT18236
Valentin-Silva A, Batalha MA, Guimarães E (2021) Disentangling the ecological basis of floral trait variation in Neotropical Piper species. Bot J Linn Soc 195:622–635. https://doi.org/10.1093/botlinnean/boaa090
de Wadt LH, O, Ehringhaus C, Kageyama PY, (2004) Genetic diversity of “Pimenta longa” genotypes (Piper spp., Piperaceae) of the Embrapa Acre germplasm collection. Genet Mol Biol 27:74–82. https://doi.org/10.1590/S1415-47572004000100013
de Oliveira Wadt LH, Kageyama PY (2004) Estrutura genética e sistema de acasalamento de Piper hispidinervum. Pesquisa Agropecuaria Brasileira 39:151–157
Watson JM, Riha K (2010) Comparative biology of telomeres: where plants stand. FEBS Lett 584:3752–3759. https://doi.org/10.1016/j.febslet.2010.06.017
Weiss H, Maluszynska J (2001) Chromosomal rearrangement in autotetraploid plants of Arabidopsis thaliana. Hereditas 133:255–261. https://doi.org/10.1111/j.1601-5223.2000.00255.x
Winterfeld G, Roser M (2007) Chromosomal localization and evolution of satellite DNAs and heterochromatin in grasses (Poaceae), especially tribe Aveneae. Plant Syst Evol 264:75–100. https://doi.org/10.1007/s00606-006-0482-1
Yuncker GT (1972) The Piperaceae of Brazil. Hoehnea 2(19–366):10024615348
Acknowledgements
The authors are financially supported by the agencies Conselho Nacional de Pesquisa e Desenvolvimento Científico (CNPq) and Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Coordenação de Apefeiçomento do Pessoal do Ensino Superior (CAPES) for the scholarship of N.R.S.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics approval
All authors read and approved the final manuscript.
Conflict of interest
The authors declare no competing interests.
Additional information
Communicated by Handling Editor: Peter Nick
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
709_2021_1721_MOESM1_ESM.jpg
Figure 1S. Illustration of high morphological similarity among the species Piper hispidinervum (a); Piper aduncum (b); Piper affinis hispidinervum (c). (JPG 155 KB)
Rights and permissions
About this article
Cite this article
Soares, N.R., Correa, C.T.R., da Silva, J.C. et al. Comparative cytogenetics of three economically important Piper L. species from the Brazilian Amazon. Protoplasma 259, 1099–1108 (2022). https://doi.org/10.1007/s00709-021-01721-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-021-01721-2