Skip to main content
SpringerLink
Log in

Optimization of somatic embryogenesis and in vitro plant regeneration of Urochloa species using picloram

  • Plant Tissue Culture
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

The establishment of an efficient methodology for regeneration in Urochloa spp. via somatic embryogenesis is an essential component of genetic engineering technology. The aim of this study was to evaluate different concentrations of growth regulators on callus induction, somatic embryogenesis, and plant regeneration of Urochloa spp. using mature seeds as the initial explant. Firstly, different concentrations (1, 2, 4, and 8 mg/L) of 2,4-D or picloram were tested using U. brizantha cv. Marandu. In a second step, the above concentrations of picloram were also used to evaluate its effect on the morphogenetic potential in different genotypes: U. brizantha cvs. Xaraés and Piatã, U. decumbens cv. Basilisk, U. humidicola cv. Llanero, and U. ruziziensis cv. Ruziziensis. There was no significant difference between concentrations of 2,4-D or picloram for the production of primary calluses in U. brizantha cv. Marandu, ranging from 58.5 to 69.2%. However, embryogenic calluses induced under picloram showed higher percentage of shoot formation after transferring to MS medium with 2 mg/L BA. A higher number of shoots were produced in mature seed cultures of U. decumbens cv. Basilisk and U. brizantha cvs. Marandu and Xaraés supplemented with 1 mg/L picloram. No albino plants were regenerated using picloram. The successful regeneration of green and morphologically normal plants opens the possibility of using this protocol to obtain transgenic plants in Urochloa spp.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  • Bai Y, Qu R (2000) An evaluation of callus induction and plant regeneration in twenty-five turf-type tall fescue (Festuca arundinace Schreb) cultivars. Grass Forage Sci 55:326–330

    Article  Google Scholar 

  • Barampuram S, Chung BY, Lee SS, An BC, Lee EM, Cho J-Y (2009) Development of an embryogenic callus induction method for centipede grass (Eremochloa ophiuroiddes Munor) and subsequent plant regeneration. In Vitro Cell Dev Biol Plant 45:155–161

    Article  Google Scholar 

  • Barro F, Martin A, Lazzeri PA, Barcelo P (1999) Medium optimization for efficient somatic embryogenesis and plant regeneration from immature inflorescences and immature scutella of elite cultivars of wheat, barley and tridordeum. Euphytica 108:161–167

    Article  Google Scholar 

  • Benson E (2000) In vitro plant recalcitrance: an introduction. In Vitro Cell Dev Biol Plant 36:141–148

    Article  Google Scholar 

  • Bregitzer P, Campbell RD (2001) Genetic markers associated with green and albino plant regeneration from embryogenic barley callus. Crop Sci 41:173–179

    Article  CAS  Google Scholar 

  • Cabral GB, Carneiro VTC, Lacerda AL, do Valle CB, Martinelli AP, Dusi DMA (2011) Somatic embryogenesis and organogenesis in apomitic and sexual Brachiaria brizantha. Plant Cell Tissue Organ Cult 107:271–282

    Article  Google Scholar 

  • Carloni E, Ribotta A, Colomba EL, Griffa S, Quiroga M, Tommasino E, Grunberg K (2014) Somatic embryogenesis from in vitro anther culture of apomitic buffel grass genotypes and analysis of regenerated plants using flow cytometry. Plant Cell Tissue Organ Cult 117:311–322

    Article  Google Scholar 

  • Colomba EL, Grunberg K, Griffa S, Ribotta A, Mroginski L, Biderbost E (2006) The effect of genotype and culture medium on somatic embryogenesis and plant regeneration from mature embryos of fourteen apomictic cultivars of buffel grass (Cenchrus ciliaris L.). Grass Forage Sci 6:2–8

    Article  Google Scholar 

  • Deo PC, Tyagi AP, Taylor M, Harding R, Becker D (2010) Factors affecting somatic embryogenesis and transformation in modern plant breeding. S Pac J Nat Appl Sci 28:27–40

    Article  Google Scholar 

  • Dhandapani M, Hong S, Aswath CR, Kim DH (2008) Regeneration of zoysia grasss (Zoysia matrella L. Merr.) cv. Konhee from young inflorescences and stem nodes. In Vitro Cell Dev Biol Plant 44:8–13

    Article  CAS  Google Scholar 

  • do Valle CB, Simioni C, Resende RMS, Jank L, Chiari L (2008) Melhoramento genético de Brachiaria. In: Resende RMS, Valle CB, Jank L (eds) Melhoramento de forrageiras tropicais. Embrapa Gado de Corte, Campo Grande, pp 13–53 (in Portuguese)

    Google Scholar 

  • Dudits D, Gyorgyey J, Bogre L, Bako L (1995) Molecular biology of somatic embryogenesis. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer Academic Publishers, Dordrecht, pp 267–308

    Chapter  Google Scholar 

  • Durzan DJ, Gupta PK (1988) Somatic embryogenesis and polyembryogenesis in conifers. In: Mizrahi A (ed) Biotechnology in Agriculture 9:53–81

  • Ferreira DF (2011) Sisvar: A computer statistical analysis system. Ciência e Agrotecnologia, Lavras 35:1039–1042

  • Gondo T, Matsumoto J, Yamakawa K, Tsuruta S, Ebina M, Akashi R (2007) Somatic embryogenesis and multiple-shoot formation from seed-derived shoot apical meristems of rhodesgrass (Chloris gayana Kunth). Grass Sci 53:138–142

    Article  Google Scholar 

  • Gray DJ (1996) Nonzygotic embryogenesis. In: Trigiano RN, Gray DJ (eds) Plant tissue culture concepts and laboratory exercises. CRC Press, New York, pp 133–147

    Google Scholar 

  • Gupta PK, Durzan DJ (1987) Biotechnology of somatic polyembryogenesis and plantlet regeneration in lobolly pine. Biotechnol 5:147–151

    Article  Google Scholar 

  • IBGE (Instituto Brasileiro De Geografia E Estatística) Sistema IBGE de recuperação automática: efetivos dos rebanhos (2013) Disponível em: http://saladeimprensa.ibge.gov.br/noticias?view=noticia&id=1&busca=1&idnoticia=22 1. Accessed 4 April 2013 (in Portuguese)

  • Ishigaki G, Gondo T, Rahman MM, Umami N, Akashi R (2014) Spontaneous appearance of polyploids in plants regenerated from embryogenic calluses derived from seedling-meristems of ruzigrass (Brachiaria ruziziensis Germain et Everard). Grass Sci 60:24–30

    Article  Google Scholar 

  • Ishigaki G, Gondo T, Suenaga K, Akashi R (2009) Multiple shoot formation, somatic embryogenesis and plant regeneration from seed-derived shoot apical meristems in ruzigrass (Brachiaria ruziziensis). Grass Sci 55:46–51

    Article  Google Scholar 

  • Ishigaki G, Gondo T, Suenaga K, Akashi R (2012) Fertile transgenic Brachiaria ruziziensis (ruzigrass) plants by particle bombardment of tetraploidized callus. J Plant Physiol 169:546–549

    Article  CAS  PubMed  Google Scholar 

  • Keller-Grein G, Maass BL, Hanson J (1996) Natural variation in Brachiaria and existing germoplasma collection. In: Miles JW, Maass BL, Valle CB (eds) Brachiaria: biology, agronomy, and improvement. Miles JW, Maass BL, do Valle CB (eds) Cali: CIAT/ Brasília: EMBRAPA-CNPGC, pp 16–42

  • Khaleghi A, Sahraroo A, Rasoulnia IN, Ataei R (2008) In vitro propagation of Alstroemeria cv. Fuego. Am Eur J Agric Environ Sci 3:492–497

    Google Scholar 

  • Komamine A, Kawahara R, Matsumoto M, Sunabori S, Toya T, Fujiwara A, Tsukahara M, Smith J, Ito M, Fukuda H, Nomura K, Fujimura T (1992) Mechanisms of somatic embryogenesis in cell cultures: physiology, biochemistry, and molecular biology. In Vitro Cell Dev Biol Plant 28:11–14

    Article  Google Scholar 

  • Lakshmanan P (2006) Somatic embryogenesis in sugarcane. In Vitro Cell Dev Biol Plant 42:201–205

    Article  CAS  Google Scholar 

  • Lenis-Manzano SJ, Araujo ACG, Valle CB, Santana EF, Carneiro VTC (2010) Histologia da embriogênese somática induzida em embriões de sementes maduras de Urochloa brizantha apomítica. Pesquisa Agropecuária Brasileira Brasília 45:435–441 (in Portuguese)

    Article  Google Scholar 

  • Macedo MCM, Kichel NA, Zimmer AH (2000) Degradação e alternativas de recuperação e renovação de pastagens. Campo Grande: EMBRAPA-CNPGC, 4 pp (Comunicado Técnico, 62) (in Portuguese)

  • Matkowoski A (2008) Plant in vitro culture for the production of antioxidants—a review. Biotechnol Adv 26:548–560

    Article  Google Scholar 

  • Miles JW, Maass BL, Valle CB (1996) Brachiaria: biology, agronomy and improvement. Centro Internacional de Agricultura Tropical–CIAT; Empresa Brasileira de Pesquisa Agropecuária–EMBRAPA, Cali; Brasília, pp 18–45

  • Minima EA, Filonova LH, Sanchez-Vera V, Suarez MF, Daniel G, Bozhkov PV (2013) Detection and measurement of necrosis in plants. Methods Mol Biol 1004:229–248

    Article  Google Scholar 

  • Mohajer S, Taha RM, Khorasani A, Yaacob JS (2012) Induction of different types of callus and somatic embryogenesis in various explants of Sainfoin (Onobrychis sativa). Aus J Crop Sci 6:1305–1313

    CAS  Google Scholar 

  • Montanari R, Lima ES, Lovera LH, Godoy LTR, Henrique AF, Dalchiavon FC (2013) Correlación de la productividad de un forraje con las propiedades físicas de un Ultisol en Aquidauana. Rev Ceres 60:102–110 (in Portuguese)

    Article  Google Scholar 

  • Moura EF, Motoike SY, Ventrella MC, Sá Júnior AQ, Carvalho M (2009) Somatic embryogenesis in macaw palm (Acrocomia aculeata) from zygotic embryos. Sci Hortic 119:447–454

    Article  CAS  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 

  • Nabors MW, Heyser JW, Dykes TA, Demon KJ (1983) Long-duration, high-frequency plant regeneration from cereal tissue cultures. Planta 157:385–391

    Article  CAS  PubMed  Google Scholar 

  • Osorio J, Calderon C, Gutierrez-Moraga A, Gidekel M (2014) The effects of growth regulators and a scanning electron microscope study of somatic embryogenesis in Antarctic hair grass (Deschampsia antarctica Desv.). Polar Biol 37:217–225

    Article  Google Scholar 

  • Ptak A, Bach A (2007) Somatic embryogenesis in tulip (Tulipa gesneriana L.) flower stem cultures. In Vitro Cell Dev Biol Plant 43:35–39

    Article  CAS  Google Scholar 

  • Scherwinski-Pereira J, Guedes RS, Fermino PC, Dilva TL, Costa FH (2010) Somatic embryogenesis and plant regeneration in oil palm using the thin cell layer technique. In Vitro Cell Dev Biol Plant 46:378–385

    Article  CAS  Google Scholar 

  • Sener O, Can E, Arslan M, Celis N (2008) Effects of genotype and Picloram concentrations on callus induction and plant regeneration from immature inflorescence of spring barley cultivars (Hordeum vulgare L.). Biotechnol Biotechnol Equip 22:915–920

    Article  CAS  Google Scholar 

  • Simioni C, Valle CB (2009) Chromosome duplication in Brachiaria (A. rich.) Stapf allows intraspecific crosses. Crop Breed Appl Biotechnol 9:328–334

    Article  Google Scholar 

  • Stefanello S, Dal Vesco LL, Ducroquet JPHJ, Nodari RO, Guerra MP (2005) Somatic embryogenesis from floral tissues of feijoa (Feijoa sellowiana Berg). Sci Hortic 105:117–126

    Article  CAS  Google Scholar 

  • Steinmacher DA, Cangahuala-Inocente GC, Clement CR, Guerra MP (2007) Somatic embryogenesis from peach palm zygotic embryos. In Vitro Cell Dev Biol Plant 43:124–132

    Article  Google Scholar 

  • Sun Y, Zang X, Jin S, Ling S, Nie Y (2003) Somatic embryogenesis and plant regeneration in wild cotton (Gossypium klotzschianum). Plant Cell Tissue Organ Cult 75:247–253

    Article  CAS  Google Scholar 

  • Sun YL, Hong SK (2009) Somatic embryogenesis and in vitro plant regeneration from various explants of the halophyte Leymus chinensis (Trin.). J Plant Biotechnol 36:236–243

    Article  Google Scholar 

  • Thome J, Palacios N, Lenis S, Roca W (1996) Applications of biotechnology to Brachiaria. In: Miles JW, Maass BL, do Valle CB (eds) Brachiaria: biology, agronomy and improvement. Cali-Colombia: CIAT; Campo Grande-Brasil: Embrapa, pp 196–204

  • Valle CB do, Savidan YH (1996) Genetics, cytogenetics and reproductive biology of Brachiaria. In: Miles JW, Maass BL, do Valle CB (eds) Brachiaria: Biology, Agronomy, and improvement. Cali: CIAT; Brasília: EMBRAPA-CNPGC. pp 147–163

  • Wang W, Zhao X, Zhuang G, Wang S, Chen F (2008) Simple hormonal regulation of somatic embryogenesis and/or shoot organogenesis in caryopsis cultures of Pogonatherum paniceum (Poaceae). Plant Cell 95:57–67

    CAS  Google Scholar 

  • Xu K, Chang Y, Liu K, Wang F, Liu ZY, Zhang T, Li TG, Zhang Y, Zhang F, Zhang J, Wang Y, Niu W, Jia S, Xie H, Tan G, Li C (2014) Regeneration of Solanum nigrum by somatic embryogenesis, involving frog egg-like body, a novel structure. Plos One 9:598–672

    Google Scholar 

  • Zhang S, Hanna W, Ozias-Akins P (2007) Comparison of callus induction and plant regeneration from different explants in triploid and tetraploid turf-type Bermuda-grasses. Plant Cell Tissue Organ Cult 70:71–78

    Article  Google Scholar 

  • Zimmerman JL (1993) Somatic embryogenesis: a model for early development in higher plants. Plant Cell 5:1411–1423

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

This study (no. 2013/04819-4) was supported by FAPESP (São Paulo State Research Foundation). LMT and LGEV are recipients of a M.Sc. scholarship from CAPES and PQ fellowship from CNPq, respectively.

Author information

Authors and Affiliations

  1. Programa de Pós-graduação em Agronomia, Laboratório de Cultura de Tecidos Vegetais, Universidade do Oeste Paulista, Rodovia Raposo Tavares, km 572, Limoeiro, 19067-175, Presidente Prudente, SP, Brazil

    Luciana Midori Takamori, Nelson Barbosa Machado Neto, Luiz Gonzaga Esteves Vieira & Alessandra Ferreira Ribas

Authors
  1. Luciana Midori Takamori
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nelson Barbosa Machado Neto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Luiz Gonzaga Esteves Vieira
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alessandra Ferreira Ribas
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alessandra Ferreira Ribas.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Takamori, L.M., Machado Neto, N.B., Vieira, L.G.E. et al. Optimization of somatic embryogenesis and in vitro plant regeneration of Urochloa species using picloram. In Vitro Cell.Dev.Biol.-Plant 51, 554–563 (2015). https://doi.org/10.1007/s11627-015-9701-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-015-9701-1

Keywords

Search

Navigation