Reducing oxidative browning during in vitro establishment of Protea cynaroides

Wu, How-Chiun; Toit, E.S. du

doi:10.1016/j.scienta.2003.07.007

Cited by 20 publications

(15 citation statements)

References 7 publications

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“…The need for a study on the action of antioxidant treatments on the in vitro culture of this plant arises from the Proteaceae being rich in phenolic compounds (Malan, 1992;Watad et al, 1992b;Rugge, 1995;Wu 794 E. Suárez et al / Span J Agric Res (2010) and du Toit, 2004). After the dissection of explants, these compounds may be oxidized by different enzymes (polyphenoloxidases and tyrosinases) which are synthesized and/or liberated due to injury during excision and sterilization of tissues (Torres, 1989).…”

Section: Discussionmentioning

confidence: 99%

“…Darkening can be reduced by limiting the production of phenolic compounds, modifying the redox potential, inactivating the phenolases, and reducing the activity of the phenolase and/or the availability of substrate (Hu and Wang, 1983). In several reports concerning Proteaceae, the use of an antioxidant solution is necessary in some phases of culture: citric and/or ascorbic acid (Pérez-Francés et al, 1995;Dias Ferreira et al, 2003;Wu and du Toit, 2004), PVP (Pérez-Francés et al, 2001a,b), or BAP applied to the entire mother plant at 200 mg L -1 . This latter significantly reduced browning and promoted bud sprouting in vitro (Rugge, 1995;Pérez-Francés et al, 2001b).…”

Section: Discussionmentioning

confidence: 99%

“…Although diverse authors have studied the micropropagation of some proteas (Hu and Wang, 1983;BenJaacov and Jacobs, 1986;Seelye et al, 1986;Rugge et al, 1989;Kunisaki, 1989Kunisaki, , 1990Bunn and Dixon, 1992a,b;Watad et al, 1992a,b;Pérez-Francés et al, 1995;Rugge, 1995;Dias Ferreira et al, 2003;Wu and du Toit, 2004), results have not been totally satisfactory, as at present there is still no firm that specialises in marketing in vitro produced protea plants. A previous report by Pérez-Francés et al (1995) showed the effect of some factors on in vitro multiplication of this cultivar.…”

Section: Introductionmentioning

confidence: 99%

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Use of multinodal explants for micropropagation of Leucadendron ‘Safari Sunset’

Suárez

Pérez-Francés

Rodríguez-Pérez

2010

Span. j. agric. res.

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Leucadendron 'Safari Sunset' is one of the most valuable Proteaceae cultivar, due to its economic importance as cutflower. In vitro tissue culture of this cultivar and others may offer a massive cloned plant production that permit to satisfy the market demand. Multinodal explants of Leucadendron 'Safari Sunset' were established in vitro on a half strength Murashige and Skoog medium supplemented with 2.2 µM benzylaminopurine (BAP). Various antioxidant treatments were used to reduce the presence of phenolic compounds in culture. No important differences in growth and multiplication between these treatments were detected during in vitro establishment and multiplication, but explants cultured on a medium with 1 g L -1 of polyvinylpyrrolidone (PVP) developed a higher number of buds in the establishment phase. Proliferation of axillary shoots was achieved with 0.88 µM BAP, being slightly higher on a medium with 2.2 µM BAP although the buds were deformed, showing abnormal leaves. The use of phytagel in this phase increased proliferation rate and shoot growth. Low percentages rooting were obtained when microcuttings were cultured on the same basal medium supplemented with several indole-3-butyric acid (IBA) concentrations. The best rooting percentage (37.5%) was obtained with 44.1 µM IBA. This work shows that micropropagation of L. 'Safari Sunset' is possible with multinodal explants, although is necessary to optimize the protocol for commercial applications.Additional key words: antioxidant treatment, in vitro tissue culture, ornamental plants, phenolic compounds, Proteaceae. ResumenEmpleo de explantos multinodales para la micropropagación de Leucadendron 'Safari Sunset' Leucadendron 'Safari Sunset' es uno de los cultivares más importantes de Proteaceae debido a su importancia económica como flor cortada. El cultivo in vitro de este cultivar y otros, puede ofrecer una producción masiva de clones que permita satisfacer la demanda del mercado. Se establecieron in vitro explantos multinodales de L. 'Safari Sunset' en un medio Murashige y Skoog, con los macronutrientes a la mitad de su concentración, y bencilaminopurina (BAP) 2,2 µM. Para reducir la presencia de compuestos fenólicos en los cultivos, se emplearon diferentes tratamientos antioxidantes. No se observaron diferencias importantes en cuanto al crecimiento y multiplicación de las yemas, entre los antioxidantes empleados durante el establecimiento y multiplicación in vitro, sin embargo, se obtuvo un mayor nú-mero de yemas (25 ± 2,82) en los explantos sembrados en un medio con 1 g L -1 de polivinilpirrolidona (PVP). La multiplicación de las yemas se logró con BAP 0,88 µM. La tasa de proliferación fue ligeramente superior al adicionar al medio de cultivo BAP 2,2 µM, pero en este medio se desarrollaron yemas deformes, con hojas anormales. El empleo de fitagel incrementó la tasa de multiplicación y crecimiento de los vástagos. Se obtuvieron bajos porcentajes de enraizamiento al cultivar los microesquejes en el medio basal suplementado con diferentes concentraci...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Use of multinodal explants for micropropagation of Leucadendron ‘Safari Sunset’

Suárez

Pérez-Francés

Rodríguez-Pérez

2010

Span. j. agric. res.

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“…In order to induce the sprouting of the microshoot on the nodal explants, the in vitro establishment of P. cynaroides nodal explants was done according to Wu and du Toit (2004): Shoot segments were taken from 1-year old P. cynaroides plants grown in black plastic bags in a greenhouse (Greenhouse temperature: 22-25°C). After removing the leaves, the explants (±1 cm, containing one to two nodes) were placed under running water for 2 h. This was followed by dipping them into 70% ethanol for 10 s and in 0.35% sodium hypochlorite for 6 min.…”

Section: Methodsmentioning

confidence: 99%

“…In vitro propagation of P. cynaroides has had limited success. In vitro establishment (Ben-Jaacov and Jacobs 1986; Wu and du Toit 2004) and in vitro multiplication (Wu 2001) have been reported. However, in vitro rooting of these microshoots has not been achieved.…”

Section: Introductionmentioning

confidence: 99%

Micrografting of Protea cynaroides

Toit

Reinhardt

2007

Plant Cell Tiss Organ Cult

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The inability to induce rooting of in vitro-established Protea cynaroides microshoots has prevented the production of complete plantlets. A successful shoot-tip micrografting technique was developed using in vitro-germinated P. cynaroides seedlings as rootstocks and axenic microshoots established from pot plants as microscions. Thirty-day old seedlings, germinated on growth-regulator-free, half-strength Murashige and Skoog medium, were decapitated and a vertical incision made from the top end. The bottom ends of microshoots established on modified Murashige and Skoog medium were cut into a wedge ('V') shape, and placed into the incision. The micrografted explants were cultured in a growth chamber with the temperature adjusted to 25 ± 2°C, with a 12-h photoperiod. Best results were obtained by placing the microscions directly onto the rootstock without any pre-treatments. Dipping the explants in anti-oxidant solution or placing a layer of medium around the graft area led to the blackening of the microscion.Abbreviations EDTA Ethylenediaminetetraacetate -BAP 6-Benzylaminopurine -GA 3

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A protocol for direct somatic embryogenesis of Protea cynaroides L. using zygotic embryos and cotyledon tissues

Toit

Reinhardt

2007

Plant Cell Tiss Organ Cult

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We describe a protocol for somatic embryogenesis of Protea cynaroides, with potential for high frequency production of this important horticultural species. Somatic embryos formed directly on both P. cynaroides mature zygotic embryos and excised cotyledons cultured on MS medium without growth regulators. The addition of growth regulators such as naphthalene acetic acid (NAA) (5; 13 and 27 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (5; 11 and 23 μM), in combination with thidiazuron (TDZ) (1 μM), benzylaminopurine (BAP) (1 μM) or kinetin (1 μM) suppressed the formation of somatic embryos. After eight weeks in culture, formation of somatic embryos was observed.Zygotic explants formed the most embryos when cultured in a 12-h photoperiod in comparison to explants cultured in the dark. Up to 83% of these embryos germinated after transferal to the germination medium containing 0.3 μM GA 3 . Significantly fewer embryos germinated in MS medium with no growth regulators, or supplemented with higher concentrations of GA 3 , while low germination percentages were also observed in MS media containing casein hydrolysate and coconut water. The germination of normal somatic embryos (two separate cotyledons and a single radicle) was observed only in media containing either no growth regulators, 0.3 μM GA 3 or 1 μM GA 3 . All embryos that germinated in high concentrations of GA 3 were malformed.Abbreviations 2,4D 2,4-Dichlorophenoxyacetic acid -BAP 6-Benzylaminopurine -GA 3 Gibberellic acid -NAA 1-Naphthalene acetic acid -PAR Photosynthetic active radiation -TDZ Thidiazuron

show abstract

Reducing oxidative browning during in vitro establishment of Protea cynaroides

Cited by 20 publications

References 7 publications

Use of multinodal explants for micropropagation of Leucadendron ‘Safari Sunset’

Use of multinodal explants for micropropagation of Leucadendron ‘Safari Sunset’

Micrografting of Protea cynaroides

A protocol for direct somatic embryogenesis of Protea cynaroides L. using zygotic embryos and cotyledon tissues

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