In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions

Cyathea atrovirens occurs in a wide range of habitats in Brazil, Paraguay, Uruguay and Argentina. In the Brazilian State of Rio Grande do Sul, this commonly found species is a target of intense exploitation, because of its ornamental characteristics. The in vitro cultura is an important tool for pro...

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Main Authors: Isabel Beatriz de Vargas, Annette Droste
Format: Article
Language:English
Published: Vicerractoría Investigación 2014-03-01
Series:Revista de Biología Tropical
Subjects:
Online Access:http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442014000200027&lng=en&tlng=en
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author Isabel Beatriz de Vargas
Annette Droste
author_facet Isabel Beatriz de Vargas
Annette Droste
author_sort Isabel Beatriz de Vargas
collection DOAJ
description Cyathea atrovirens occurs in a wide range of habitats in Brazil, Paraguay, Uruguay and Argentina. In the Brazilian State of Rio Grande do Sul, this commonly found species is a target of intense exploitation, because of its ornamental characteristics. The in vitro cultura is an important tool for propagation which may contribute toward the reduction of extractivism. However, exogenous contamination of spores is an obstacle for the success of aseptic long-term cultures. This study evaluated the influence of different sterilization methods combined with storage conditions on the contamination of the in vitro cultures and the gametophytic development of C. atrovirens, in order to establish an efficient propagation protocol. Spores were obtained from plants collected in Novo Hamburgo, State of Rio Grande do Sul, Brazil. In the first experiment, spores stored at 7oC were surface sterilized with 0.5, 0.8 and 2% of sodium hypochlorite (NaClO) for 15 minutes and sown in Meyer’s culture medium. The cultures were maintained in a growth room at 26±1ºC for a 12-h photoperiod and photon flux density of 100μmol/m²/s provided by cool white fluorescent light. Contamination was assessed at 60 days, and gametophytic development was scored at 30, 60, 120 and 130 days of in vitro culture, analyzing 300 individuals for each treatment. There was no significant difference in culture contamination among the different sodium hypochlorite concentrations tested, and all treatments allowed for the development of cordiform gametophytes at 130 days of culture. In the second experiment, spores stored at 7 and -20°C were divided into two groups. Half of the spores were surface sterilized with 2% of NaClO for 15 minutes and the other half was not sterilized. All spores were sown in Meyer’s medium supplemented with one of the following antibiotics: nystatin, Micostatin® and actidione. The culture conditions and the procedures used for evaluating contamination and gametophytic development were the same described for the first experiment. No contamination was observed in spores stored at -20°C and treated with NaClO and actidione. In all treatments, cordiform gametophytes presenting antheridia were observed at 120 days. The percentages of these gametophytes increased from 120 to 130 days and no significant differences were observed among treatments. Archegonia were observed on cordiform gametophytes at 130 days. The findings provide data relevant to in vitro propagation procedures of this species, which may increase the availability of plants for ornamental purposes, therefore contributing to the reduction of the exploitation of endangered tree ferns species. Rev. Biol. Trop. 62 (1): 299-308. Epub 2014 March 01.
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spelling doaj.art-ee987369d9f64aaf83b1237910fe6ac22023-09-02T19:26:40ZengVicerractoría InvestigaciónRevista de Biología Tropical0034-77442014-03-01621359368S0034-77442014000200027In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditionsIsabel Beatriz de Vargas0Annette Droste1Universidade FeevaleUniversidade FeevaleCyathea atrovirens occurs in a wide range of habitats in Brazil, Paraguay, Uruguay and Argentina. In the Brazilian State of Rio Grande do Sul, this commonly found species is a target of intense exploitation, because of its ornamental characteristics. The in vitro cultura is an important tool for propagation which may contribute toward the reduction of extractivism. However, exogenous contamination of spores is an obstacle for the success of aseptic long-term cultures. This study evaluated the influence of different sterilization methods combined with storage conditions on the contamination of the in vitro cultures and the gametophytic development of C. atrovirens, in order to establish an efficient propagation protocol. Spores were obtained from plants collected in Novo Hamburgo, State of Rio Grande do Sul, Brazil. In the first experiment, spores stored at 7oC were surface sterilized with 0.5, 0.8 and 2% of sodium hypochlorite (NaClO) for 15 minutes and sown in Meyer’s culture medium. The cultures were maintained in a growth room at 26±1ºC for a 12-h photoperiod and photon flux density of 100μmol/m²/s provided by cool white fluorescent light. Contamination was assessed at 60 days, and gametophytic development was scored at 30, 60, 120 and 130 days of in vitro culture, analyzing 300 individuals for each treatment. There was no significant difference in culture contamination among the different sodium hypochlorite concentrations tested, and all treatments allowed for the development of cordiform gametophytes at 130 days of culture. In the second experiment, spores stored at 7 and -20°C were divided into two groups. Half of the spores were surface sterilized with 2% of NaClO for 15 minutes and the other half was not sterilized. All spores were sown in Meyer’s medium supplemented with one of the following antibiotics: nystatin, Micostatin® and actidione. The culture conditions and the procedures used for evaluating contamination and gametophytic development were the same described for the first experiment. No contamination was observed in spores stored at -20°C and treated with NaClO and actidione. In all treatments, cordiform gametophytes presenting antheridia were observed at 120 days. The percentages of these gametophytes increased from 120 to 130 days and no significant differences were observed among treatments. Archegonia were observed on cordiform gametophytes at 130 days. The findings provide data relevant to in vitro propagation procedures of this species, which may increase the availability of plants for ornamental purposes, therefore contributing to the reduction of the exploitation of endangered tree ferns species. Rev. Biol. Trop. 62 (1): 299-308. Epub 2014 March 01.http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442014000200027&lng=en&tlng=enactidionaantibióticocultivo in vitroesterilización superficialgametófitogerminaciónhelecho arborescente
spellingShingle Isabel Beatriz de Vargas
Annette Droste
In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions
Revista de Biología Tropical
actidiona
antibiótico
cultivo in vitro
esterilización superficial
gametófito
germinación
helecho arborescente
title In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions
title_full In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions
title_fullStr In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions
title_full_unstemmed In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions
title_short In vitro propagation of Cyathea atrovirens (Cyatheaceae): spore storage and sterilization conditions
title_sort in vitro propagation of cyathea atrovirens cyatheaceae spore storage and sterilization conditions
topic actidiona
antibiótico
cultivo in vitro
esterilización superficial
gametófito
germinación
helecho arborescente
url http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442014000200027&lng=en&tlng=en
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