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In vitro plant regeneration of Passiflora setacea D.C. (Passifloraceae): the influence of explant type, growth regulators, and incubation conditions

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Abstract

The present study aimed to establish a protocol for in vitro organogenesis of Passiflora setacea and to determine the genetic stability of regenerated plants. Three types of explants (leaf, hypocotyl, and root), four growth regulator combinations [Murashige and Skoog (MS) salts, MS + 6-benzyladenine (BA), MS + thidiazuron (TDZ), and MS + BA + TDZ], and two light regimes (16-h photoperiod and continuous darkness) were tested. After 30 d on induction medium, the percentage of explants forming shoots was evaluated. Direct and indirect organogenesis was evident from hypocotyl- and root-derived explants, whereas only indirect organogenesis was observed from leaf explants. The presence of BA was essential for shoot formation from leaf explants and improved the response of hypocotyl segments under a 16-h photoperiod compared to the cytokinin-free control. However, after transfer to shoot elongation medium, the greatest number of elongated shoots was obtained from hypocotyl segments that had been induced on BA + TDZ medium under a 16-h photoperiod, as was also observed for root explants. Flow cytometry analysis confirmed the genetic stability of the regenerants based on DNA quantity (2C = 2.57 pg) in comparison with seed-derived plantlets (2C = 2.60 pg). This is the first report on the in vitro regeneration of P. setacea.

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References

  • Appezzato-da-Glória B, Vieira MLC, Dornelas MC (1999) Anatomical studies of in vitro morphogenesis in leaf explants of passionfruit. Pesq Agropec Bras 34:2007–2013

    Article  Google Scholar 

  • Braga MF, Santos EC, Junqueira NTV, Sousa AATC, Faleiro FG, Rezende LN, Junqueira KP (2006) Enraizamento de estacas de três espécies silvestres de Passiflora. Rev Bras Frutic 28:284–288

    Article  Google Scholar 

  • Dias LLC, Santa-Catarina C, Ribeiro DM, Barros RS, Floh ELS, Otoni WC (2009) Ethylene and polyamine production patterns during in vitro shoot organogenesis of two passion fruit species as affected by polyamines and their inhibitor. Plant Cell Tissue Organ Cult 99:199–208

    Article  CAS  Google Scholar 

  • Dias LLC, Ribeiro DM, Santa-Catarina C, Barros RS, Floh EIS, Otoni WC (2010) Ethylene and polyamine interactions in morphogenesis of Passiflora cincinnata: effects of ethylene biosynthesis and action modulators, as well as ethylene scavengers. Plant Growth Regul 62:9–19

    Article  CAS  Google Scholar 

  • Doležel J, Binarová P, Lucretti S (1989) Analysis of nuclear DNA content in plant cells by flow cytometry. Biol Plant 31:113–120

    Article  Google Scholar 

  • Doležel J, Greilhuber J, Suda J (2007) Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc 2:2233–2244

    Article  PubMed  Google Scholar 

  • Dornelas MC, Vieira MLC (1994) Tissue culture studies on species of Passiflora. Plant Cell Tissue Organ Cult 36:211–217

    Article  CAS  Google Scholar 

  • Escobedo-GraciaMedrano RM, Maldonado-Borges JI, Burgos-Tan MJ, Valadez-González N, Ku-Caiuch JR (2014) Using flow cytometry and cytological analyses to assess the genetic stability of somatic embryo-derived plantlets from embryogenic Musa acuminata Colla (AA) ssp. malaccensis cell suspension cultures. Plant Cell Tissue Organ Cult 116:175–185

    Article  CAS  Google Scholar 

  • Evans DA, Sharp WR, Medina-Filho HP (1984) Somaclonal and gametoclonal variation. Am J Bot 77:759–774

    Article  Google Scholar 

  • Faleiro FG, Junqueira NTV, Fávero AP, Lopes MA (2008) Pré-melhoramento de plantas: experiências de sucesso. In: Faleiro FG, Farias Neto AL, Ribeiro Junior WQ (eds) Pré-melhoramento, melhoramento e pós-melhoramento: Estratégias e desafios. Embrapa Cerrados, Planaltina, pp 43–62

    Google Scholar 

  • Fernando JA, Vieira MLC, Machado SR, Appezzato-da-Glória B (2007) New insights into the in vitro organogenesis process: the case of Passiflora. Plant Cell Tissue Organ Cult 91:37–44

    Article  Google Scholar 

  • Flores PS, Otoni WC, Dhingra OD, Souza Diniz SPS, Santos TM, Bruckner CH (2012) In vitro selection of yellow passion fruit genotypes for resistance to Fusarium vascular wilt. Plant Cell Tissue Organ Cult 108:37–45

    Article  CAS  Google Scholar 

  • Fratini R, Ruiz MN (2002) Comparative study of different cytokinins in the induction of morphogenesis in lentil (Lens culinaris Medik.). In vitro Cell Dev Biol Plant 38:46–51

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  CAS  PubMed  Google Scholar 

  • Garcia R, Pacheco G, Falcão E, Borges G, Mansur E (2011) Influence of type of explant, plant growth regulators, salt composition of basal medium, and light on callogenesis and regeneration in Passiflora suberosa L. (Passifloraceae). Plant Cell Tissue Organ Cult 106:47–54

    Article  CAS  Google Scholar 

  • George EF, Hall MA, DeKlerk GJ (2008) Plant propagation by tissue culture. Volume 1: The Background, 3rd edn. Springer, Dordrecht

    Google Scholar 

  • Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 27:137–138

    Google Scholar 

  • Karp A (1995) Somaclonal variation as a tool for crop improvement. Euphytica 85:295–302

    Article  Google Scholar 

  • Lombardi SP, Passos IRS, Nogueira MCS, Appezzato-da-Glória B (2007) In vitro shoot regeneration from roots and leaf discs of Passiflora cincinnata Mast. Braz Arch Biol Technol 50:239–247

    Article  Google Scholar 

  • Loureiro J, Pinto G, Lopes T, Doležel J, Santos C (2005) Assessment of ploidy stability of the somatic embryogenesis process in Quercus suber L. using flow cytometry. Planta 221:815–822

    Article  CAS  PubMed  Google Scholar 

  • Manica I, Brancher A, Sanzonowicz C, Icuma IM, Aguiar JLP, Azevedo JA, Vasconcellos MAS, Junqueira NTV (2005) Maracujá-doce: Tecnologia de produção, pós-colheita, mercado, 1ªth edn. Cinco Continentes, Porto Alegre

    Google Scholar 

  • Monteiro ACBA, Nakasawa GT, Mendes BMJ, Rodriguez APM (2000) Regeneração in vitro de Passiflora suberosa a partir de discos foliares. Sci Agric 57:571–573

    Article  CAS  Google Scholar 

  • Moran-Robles MJ (1979) Potentiel morphogénétique dês entrenoeuds de Passiflora edulis var. flavicarpa Deg. et P. mollissima Bailey en culture in vitro. Turrialba 29:224–228

    Google Scholar 

  • Moreira-Dias JM, Molina RV, Bordón Y, Guardiola JL, Garcia-Luiz A (2000) Direct and indirect shoot organogenic pathways in epicotyl cuttings of Troyer citrange differ in hormone requirements and in their response to light. Ann Bot 85:103–110

    Article  CAS  Google Scholar 

  • Müller D, Leyser O (2011) Auxin, cytokinin and the control of shoot branching. Ann Bot 107:1203–1212

    Article  PubMed Central  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nhut DT, Khiet BLT, Thi NN, Thuy DTT, Duy N, Hai NT, Huyen PX (2007) High frequency shoot formation of yellow passion fruit (Passiflora edulis f. flavicarpa) via thin cell layer (TCL) technology. In: Jain SM, Haggman H (eds) Protocols for micropropagation of woody trees and fruits. Springer, Dordrecht, pp 417–426

    Chapter  Google Scholar 

  • O’Brien TP, McCully ME (1981) The study of plant structure principles and selected methods, 1st edn. Termarcarphi Pty, Melbourne

    Google Scholar 

  • Oliveira JC, Ferreira FR (1991) Melhoramento genético do maracujazeiro. In: São José AR, Ferreira FR, Vaz RL (eds) A cultura do maracujá no Brasil. FUNEP, Jaboticabal, pp 211–246

    Google Scholar 

  • Oliveira JC, Ruggiero C (2005) Espécies de maracujá com potencial agronômico. In: Faleiro FG, Junqueira NTV, Braga MF (eds) Maracujá: germoplasma e melhoramento genético. Embrapa Cerrados, Planaltina, pp 142–158

    Google Scholar 

  • Otoni WC, Paim Pinto DL, Rocha DI, Vieira LM, Dias LLC, Silva ML, Silva CV, Lani ERG, Silva LC, Tanaka FAO (2013) Organogenesis and somatic embryogenesis in passionfruit (Passiflora sps.). In: Aslam J, Srivastava OS, Sharma MP (eds) Somatic embryogenesis and gene expression. Narosa Publishing House, New Delhi, pp 1–17

    Google Scholar 

  • Pacheco G, Garcia R, Lugatto D, Vianna M, Mansur E (2012) Plant regeneration, callus induction and establishment of cell suspension cultures of Passiflora alata Curtis. Sci Hortic 144:42–47

    Article  CAS  Google Scholar 

  • Pádua JG, Schwingel LC, Mundin RC, Salomão AN, Roverijosé SCB (2011) Germinação de sementes de Passiflora setacea e dormência induzida pelo armazenamento. Rev Bras Sementes 33:80–85

    Article  Google Scholar 

  • Paim Pinto DL, Barros BA, Viccini LF, Campos JMS, Silva ML, Otoni WC (2010) Ploidy stability of somatic embryogenesis-derived Passiflora cincinnata Mast plants as assessed by flow cytometry. Plant Cell Tissue Organ Cult 103:71–79

    Article  Google Scholar 

  • Pinto APC, Monteiro-Hara ACBA, Stipp LCL, Mendes BMJ (2010) In vitro organogenesis of Passiflora alata. In Vitro Cell Dev Biol Plant 46:28–33

    Article  Google Scholar 

  • Reis LB, Silva ML, Lima ABP, Oliveira MLP, Pinto DLP, Lani ERG, Otoni WC (2007) Agrobacterium rhizogenes-mediated transformation of passionfruit species: Passiflora cincinnata and P. edulis flavicarpa. Acta Hortic 738:425–431

    Google Scholar 

  • Rocha DI, Vieira LM, Tanaka FAO, Silva LC, Otoni WC (2012a) Anatomical and ultrastructural analyses of in vitro organogenesis from root explants of commercial passion fruit (Passiflora edulis Sims). Plant Cell Tissue Organ Cult 111:69–78

    Article  CAS  Google Scholar 

  • Rocha DI, Vieira LM, Tanaka FA, Silva LC, Otoni WC (2012b) Somatic embryogenesis of a wild passion fruit species Passiflora cincinnata Masters: histocytological and histochemical evidences. Protoplasma 249:747–758

    Article  PubMed  Google Scholar 

  • Rosa YBJ, Dornelas MC (2012) In vitro plant regeneration and de novo differentiation of secretory trichomes in Passiflora foetida L. (Passifloraceae). Plant Cell Tissue Organ Cult 108:91–99

    Article  CAS  Google Scholar 

  • SAS Institute (2010) SAS/STAT® 9.1 Users guide. SAS Institute Inc, Cary

    Google Scholar 

  • Siegien I, Adamczuk A, Wróblewska K (2013) Light affects in vitro organogenesis of Linum usitatissimum L. and its cyanogenic potential. Acta Physiol Plant 35:781–789

    Article  CAS  Google Scholar 

  • Silva ML, Paim Pinto DL, Guerra MP, Floh EIS, Bruckner CH, Otoni WC (2009) A novel regeneration system for wild passion fruit species (Passiflora cincinnata Mast.) based on somatic embryogenesis from mature zygotic embryos. Plant Cell Tiss Organ Cult 99:47–54

  • Silva CV, Oliveira LS, Loriato VAP, Silva LC, Campos JMS, Viccini LF, Oliveira EJ, Otoni WC (2011) Organogenesis from root explants of commercial populations of Passiflora edulis Sims and a wild passionfruit species, P. cincinnata Masters. Plant Cell Tissue Organ Cult 107:407–416 

    Article  Google Scholar 

  • Smulders MJM, de Klerk GJ (2011) Epigenetics in plant tissue culture. Plant Growth Regul 63:137–146

    Article  CAS  Google Scholar 

  • Soares WS, Rêgo MM, Rêgo ER, Barroso PA, Nascimento KS, Ferreira KT (2012) Estabelecimento in vitro e micropropagação de maracujá silvestre (Passiflora foetida L.). Rev Bras Plantas Med 14:138–142

    Article  CAS  Google Scholar 

  • Souza MM, Palomino G, Pereira TNS, Pereira MG, Viana AP (2004) Flow cytometric analysis of genome size variation in some Passiflora species. Hereditas 141:131–138

    Article  Google Scholar 

  • Trevisan F, Mendes BMJ (2005) Optimization of in vitro organogenesis in passion fruit (Passiflora edulis f. flavicarpa). Sci Agricola 62:346–350

    Article  Google Scholar 

  • Trevisan F, Mendes BMJ, Maciel SC, Vieira MLC, Meletti LMM, Rezende JAM (2006) Resistance to Passion fruit woodness virus in transgenic passionflower expressing the virus coat protein gene. Plant Dis 90:1026–1030

    Article  CAS  Google Scholar 

  • Vieira MLC, Carneiro MS (2004) Passiflora spp., passionfruit. In: Litz RE (ed) Biotechnology of fruit and nut crops. CABI Publishing, Wallingford, pp 435–453

    Google Scholar 

  • Zerbini FM, Otoni WC, Vieira MLC (2008) Transgenic passionfruit. In: Kole C, Hall TC (eds) Compendium of transgenic crop plants: tropical and subtropical fruits and nuts, 1st edn. John Wiley and Sons, Berlin, pp 213–234

    Google Scholar 

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Acknowledgments

The authors would like to thank Prof. Francisco Tanaka for use of the electron microscope facility at NAP/MEPA-ESALQ/USP, CAPES (Brasília, DF, Brazil), CNPq (Brasília, DF, Brazil), and FAPEMIG (Grant CRA-APQ-01451-12; Belo Horizonte, MG, Brazil). We also acknowledge Prof. J. Doležel for making available L. esculentum ‘Stupicke.’

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Authors and Affiliations

  1. Plant Biology Department, Federal University of Viçosa, University Campus, Peter Henry Rolfs Avenue, 36570-900, Viçosa, MG, Brazil

    Lorena Melo Vieira, Diego Ismael Rocha, Mariana Futia Taquetti, Luzimar Campos da Silva & Wagner Campos Otoni

  2. Institute of Biological Sciences, Department of Biology, Federal University of Juiz de Fora, 36036-900, Juiz de Fora, MG, Brazil

    José Marcello Salabert de Campos & Lyderson Faccio Viccini

  3. Departamento de Biologia Vegetal/BIOAGRO, Universidade Federal de Viçosa, Av. P. H. Rolfs s/n, Ed. CCBII, Campus Universitário, 36570-900, Viçosa, MG, Brazil

    Wagner Campos Otoni

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  1. Lorena Melo Vieira
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  2. Diego Ismael Rocha
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  4. Luzimar Campos da Silva
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Correspondence to Wagner Campos Otoni.

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Vieira, L.M., Rocha, D.I., Taquetti, M.F. et al. In vitro plant regeneration of Passiflora setacea D.C. (Passifloraceae): the influence of explant type, growth regulators, and incubation conditions. In Vitro Cell.Dev.Biol.-Plant 50, 738–745 (2014). https://doi.org/10.1007/s11627-014-9650-0

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