Effective medium for in vitro sprouting of the buds and multiplication of the plantlets, and identification of the fungal contaminant associated with the explants of Gnetum

Abwe Mercy Ngone 1, *, Lawrence Monah Ndam 1, Rita Mungfu Njilar 1, Doungous Oumar 2 and Thomas Eku Njock 1

1 Department of Agronomic and Applied Molecular Sciences, University of Buea, PO Box 63 Buea, Cameroon.
2 Jay P. Johnson Biotechnology Laboratory, Ekona Regional Research Centre, Institute of Agricultural Research for Development (IRAD), PMB 25, Buea, Cameroon.
 
Research Article
GSC Biological and Pharmaceutical Sciences, 2021, 16(02), 001–013.
Article DOI: 10.30574/gscbps.2021.16.2.0221
Publication history: 
Received on 30 June 2021; revised on 04 August 2021; accepted on 06 August 2021
 
Abstract: 
Plant tissue culture requires the optimization of growth media. Gnetum, known locally in Cameroon as “Eru” is an indigenous gymnospermous vegetable with diverse medicinal, nutritional, cultural and socio-economic values. This resource is over-exploited and expected to neighboring countries, resulting to increased scarcity in the forest. Preliminary work on the in vitro culture of nodal cuttings was faced by the problem of fungal contamination. It was therefore necessary to isolate and identify the fungal contaminant, optimize the surface sterilization of field material and compose an appropriate medium for sprouting.
Pure cultures of the fungus were obtained and grown on Potato Dextrose Agar (PDA) and Sabouraud Dextrose Agar (SDA). The identification was based on the appearance of the fungal growth on plates and also on the microscopic view. This was affected by the use of keys. Gnetum explants were disinfected with the various concentrations of disinfectants, preceded in some instances by pre-treatments, as well as incorporating fungicides in the culture medium. Two different culture media were employed: the Woody Plant Medium (WPM) and the Murashige and Skoog (MS) based establishment medium (Y-1).
Gnetum was found to live in association with a complex of Microsporum species. The level of contamination of cultures was reduced from 100% to 40% when pre-treated before disinfection and even lower to 10% by incorporating fungicides in the medium. Sprouting was observed in WPM.
This study provides baseline information on the in vitro propagation of Gnetum and thus opens up avenues for more research to be carried out in this field.
 
Keywords: 
Micropropagation; Gnetum; Microsporum species; Sterilisation; Biotechnology
 
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