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Direct organogenesis from hypocotyl-derived explants of annatto (Bixa orellana)

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Abstract

The relative importance of explant, cytokinin type, carbon source and gelling agent for annatto organogenesis was studied. The best organogenic response, including adventitious shoot number and elongation, was obtained when hypocotyl segments and rooted hypocotyls were cultured onto MS medium supplemented with 4.56 μM zeatin, 87.6 mM sucrose, and 2.8 g l−1 Phytagel®. Adventitious shoots derived from hypocotyl segments were less frequent and more difficult to elongate than those derived from rooted hypocotyls. Thidiazuron (TDZ) promoted a higher organogenic response in rooted hypocotyls, resembling a rosette-like structure, but impaired shoot elongation. Histological investigation showed that zeatin-induced meristemoids originated mainly from wounding tissues, and that TDZ induced a high level of mitotic division resulting in several proliferation zones nearby the epidermis and outer cortical tissues. Rhizogenesis efficiency (rooting frequency and root number) was greater at the highest indole-3-butyric acid (IBA) concentration (5.0 μM) employed, although calli occurred at the basal end of shoots. Eighty percent of rooted plantlets survived after acclimatization. This optimized regeneration protocol may enable further development of an efficient genetic transformation protocol for this species.

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References

  • Al-Jaboory KH, Skirvin RM & Williams DJ (1998) Callus induc-tion and adventitious shoot regeneration of gardenia (Gardenia jasminoides Ellis). Sci. Hort. 72: 171–178

    Google Scholar 

  • Biahoua A & Bonneau L (1999) Control of in vitro somatic embryogenesis of the spindle tree (Euonymus europaeus L.) by the sugar type and the osmotic potential of the culture medium. Plant Cell Rep. 19: 185–190

    Google Scholar 

  • Borkowska B & Szezebra J (1991) Influence of different carbon sources on invertase-activity and growth of sour cherry (Prunus cerasus L.) shoot cultures. J. Exp. Bot. 42: 911–915

    Google Scholar 

  • Buter B, Pescitelli SM, Berger K, Schmid JE & Stamp P (1993) Autoclaved and filter sterilized liquid media in maize anther culture: significance of activated charcoal. Plant Cell Rep. 13: 79–82

    Google Scholar 

  • D'souza MC & Sharon M (2001) In vitro clonal propagation of annatto (Bixa orellana L.). In Vitro Cell. Dev. Biol. - Plant 37: 168–172

    Google Scholar 

  • Eapen S, Tivarekar S & George L (1998) Thidiazuron-induced shoot regeneration in pigeonpea (Cajanus cajan L.). Plant Cell Tiss. Org. Cult. 53: 217–220

    Google Scholar 

  • Fuentes SRL, Calheiros MBP, Manetti-Filho J & Vieira LGE (2000) The effects of silver nitrate and different carbohydrate sources on somatic embryogenesis in Coffea canephora. Plant Cell Tiss. Org. Cult. 60: 5–13

    Google Scholar 

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

    Google Scholar 

  • García-Luis A, Bórdon Y, Moreira-Dias JM, Molina RV & Guardiola JL (1999) Explant orientation and polarity determine the morphogenic response of epicotyl segments of Troyer citrange. Ann. Bot. 84: 715–723

    Google Scholar 

  • Henry BS (1996) Natural food colours. In: Hendry GAF & Houg-hton JD (eds) Natural Food Colorants (pp. 0–79). Blackie Academic Professional, Glasgow

    Google Scholar 

  • Huang LC, Kohashi C, Vangundy R & Murashige T (1995) Effects of common components of hardness of culture media prepared with gelrite. In Vitro Cell. Dev. Biol. - Plant 31: 84–89

    Google Scholar 

  • Huetteman CA & Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tiss. Org. Cult. 33: 105–119

    Google Scholar 

  • Johansen DA (1940) Plant Microtechnique. McGraw-Hill, New York

    Google Scholar 

  • Kärkönen A (2000) Anatomical study of zygotic and somatic embryos of Tilia cordata. Plant Cell Tiss. Org. Cult. 61: 205–214

    Google Scholar 

  • Klimaszewska K, Bernier-Cardou M, Cyr DR & Sutton BCS (2000) Influence of gelling agents on medium gel strength, water availability, tissue water potential, and maturation response in embryogenic cultures of Pinus strobus L. In Vitro Cell. Dev. Biol. - Plant 36: 279–286

    Google Scholar 

  • Kotsias D & Roussos PA (2001) An investigation on the effect of different plant growth regulating compounds in in vitro shoot tip and node culture of lemon seedlings. Sci. Hort. 89: 115–128

    Google Scholar 

  • Ladyman JAR & Girard B (1992) Cucumber somatic embryo development on various gelling agents and carbohydrate sources. Hort. Sci. 27: 164–165

    Google Scholar 

  • Leonardi C, Ruggeri A & Malfa S (2001) Hormone effects on in vitro proliferation and rooting of Grevillea explants. Sci. Hort. 90: 335–341

    Google Scholar 

  • Marks TM & Simpson SE (2000) Interaction of explant type and indole-3-butyric acid during rooting in vitro in a range of difficult and easy-to-root woody plants. Plant Cell Tiss. Org. Cult. 62: 65–74

    Google Scholar 

  • Mensuali-Sodi A, Panizza M & Tognoni F (1992) Quantification of ethylene losses in different container-seal systems and com-parison of biotic and abiotic contributions to ethylene accumula-tion in cultured tissues. Physiol. Plant. 84: 472–476

    Google Scholar 

  • Muday GK & DeLong A (2001) Polar auxin transport: controlling where and how much. Trends Plant Sci. 6: 535–542

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 18: 473–497

    Google Scholar 

  • Murthy BNS & Saxena PK (1998) Somatic embryogenesis and plant regeneration of Neem (Azadirachta indica A. Juss). Plant Cell Rep. 17: 469–475

    Google Scholar 

  • Neštákova M, Havrlentová M & Faragó J (2000) Effect of gelling agents on in vitro multiplication of two ornamental plants. Biologia, Bratislava 55: 409–411

    Google Scholar 

  • Pérez-Molphe-Balch E & Ochoa-Alejo N (1997) In vitro plant regeneration of Mexican lime and mandarin by direct or-ganogenesis. Hort. Sci. 32: 931–934

    Google Scholar 

  • Pérez-Tornero O, Egea J, Olmos E & Burgos L (2001) Control of hyperhydricity in micropropagated apricot cultivars. In Vitro Cell. Dev. Biol. - Plant 37: 250–254

    Google Scholar 

  • Petersen KK, Hansen J & Krogstrup P (1999) Significance of different carbon sources and sterilization methods on callus induction and plant regeneration of Miscanthus×ogiformis Honda 'Giganteus'. Plant Cell Tiss. Org. Cult. 58: 189–197

    Google Scholar 

  • Pritchard J, Wyn-Jones RG & Tomos AD (1991) Turgor, growth and rheological gradients in wheat roots following osmotic stress. J. Exp. Bot. 42: 1043–1049

    Google Scholar 

  • Pua EC (1999) Morphogenesis in cell and tissue cultures role of ethylene and polyamines. In: Soh W-Y & Bhojwani SS (eds) Morphogenesis in Plant Tissue Cultures (pp. 255–303). Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Scherer RA, Muller E, Lippert H & Wolff G (1988) Multielement analysis of agar and gelrite impurities investigated by inductively coupled plasma emission spectrometry as well as physical properties of tissue culture media prepared with agar and gellan gum gelrite. Acta Hort. 226: 655–658

    Google Scholar 

  • Scholten HJ & Pierik RLM (1998) Agar as a gelling agent: differential biological effects in vitro. Sci. Hort. 77: 109–116

    Google Scholar 

  • Sharma P & Rajam MV (1995) Genotype, explant and position effects on organogenesis and somatic embryogenesis in eggplant (Solanum melongena L.). J. Exp. Bot. 46: 135–141

    Google Scholar 

  • Sul I & Korban SS (1998) Effects of media, carbon sources and cytokinins on shoot organogenesis in the Christmas tree Scots pine (Pinus sylvestris L.). J. Hort. Sci. Biotech. 73: 822–827

    Google Scholar 

  • Tonon G, Capuano M & Di Marco A (2001) Plant regeneration of Fraxinus angustifolia by in vitro shoot organogenesis. Sci. Hort. 87: 291–301

    Google Scholar 

  • Tsuro M, Koda M & Inoue M (1999) Comparative effect of different types of cytokinin for shoot formation and plant regeneration in leaf-derived callus of lavender (Lavandula vera DC). Sci. Hort. 81: 331–336

    Google Scholar 

  • Valera-Montero LL & Ochoa-Alejo N (1992) A novel approach for chili pepper (Capsicum annuum L.) plant regeneration: shoot induction in rooted hypocotyls. Plant Sci. 84: 215–219

    Google Scholar 

  • Vieira IMS, Barbosa ASA, Serra AGP, Silva SPG, Mota MGC & Botelho MN (2000) Embriogênese somática de urucu (Bixa o orellana), cultivar EMBRAPA-37. Proceedings of 51° Congresso Brasileiro de Botânica. Brasília (p. 23)

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Correspondence to Wagner Campos Otoni.

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Borges de Paiva Neto, V., Ribeiro da Mota, T. & Campos Otoni, W. Direct organogenesis from hypocotyl-derived explants of annatto (Bixa orellana). Plant Cell, Tissue and Organ Culture 75, 159–167 (2003). https://doi.org/10.1023/A:1025063906822

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