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; Lawrence Monah  Ndam; Rita Mungfu  Njilar; Doungous  Oumar; Thomas Eku  Njock

doi:10.30574/gscbps.2021.16.2.0221

Authors

Abwe Mercy Ngone Department of Agronomic and Applied Molecular Sciences, University of Buea, PO Box 63 Buea, Cameroon.
Lawrence Monah Ndam Department of Agronomic and Applied Molecular Sciences, University of Buea, PO Box 63 Buea, Cameroon.
Rita Mungfu Njilar Department of Agronomic and Applied Molecular Sciences, University of Buea, PO Box 63 Buea, Cameroon.
Doungous Oumar Jay P. Johnson Biotechnology Laboratory, Ekona Regional Research Centre, Institute of Agricultural Research for Development (IRAD), PMB 25, Buea, Cameroon.
Thomas Eku Njock Department of Agronomic and Applied Molecular Sciences, University of Buea, PO Box 63 Buea, Cameroon.

DOI:

https://doi.org/10.30574/gscbps.2021.16.2.0221

Keywords:

Micropropagation, Gnetum, Microsporum species, Sterilisation, Biotechnology

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

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References

Oumar D, Minyaka E, Medza-Mve SD, Medueghue AF, Ngone AM, Simo C, Nsimi AM. Improving propagation methods of Gnetum africanum and G. buchholzianum from cuttings for rapid multiplication, domestication and conservation. Agroforestry Systems. 2019; 93: 1557-1565.

Shiembo PN. Domestication of Gnetum spp by vegetative propagation techniques. In: R.R. Schippers and L. Budd (eds.) African Indigenous Vegetables, workshop Proceedings. Limbe, Cameroon. IPGRI and NRI. 1997; 31-35.

Biye EH, Balkwill K, Cron GV. A clarification of Gnetum L. (Gnetaceae) in Africa and the description of two new species. Plant Syst Evol. 2014; 300: 263–272.

Iloh AC, Isu NR, Kuta DD. In vitro callus initiation of a ‘threatened’ Nigerian leafy vegetable, Gnetum africanum (WILW). International Journal of Biodiversity and Conservation. 2009; 1(4): 081-086

Clark LE, Asha S, Ndam N, Blackmore Eru Gnetum africanum and G. buchholziannum. In: Clark LE, Sunderland TC (eds) The key non-timber forest products of central Africa: state of the knowledge. US Agency for International Development, Office of Sustainable Development, Bureau for Africa, Washington, DC. 2004; 19-22.

Ingram V, Ndumbe LN, Ewane, ME. Small Scale, High Value: Gnetum africanum and buchholzianum Value Chains in Cameroon. Small-scale Forestry. 2012; 11: 539–556.

Schippers RR. African Indigenous Vegetables. An Overview of the Cultivated Species. Natural Resources Institute/ACP-EU. Technical Center for Agricultural and Rural Cooperation, Chatham, UK. 2000; 210.

Fadi A, Mafu AA, Carole R. Gnetum africanum: A Wild Food Plant from the African Forest with Many Nutritional and Medicinal Properties. Journal of Medicinal Food. 2011; 14 (11): 289–1297.

Tchoundjeu Z, Mpeck M, Asaah E, Amougou A. The role of vegetative propagation in the domestication of Pausinystalia johimbe (K. Schum), a highly threatened medicinal species of West and Central Africa. Forest Ecology and Management. 2003; 188(1): 175-183.

Nkefor J, Ndam N, Blackmore P, Engange F, Monono C.Transfer of eru (Gnetum africanum Welw. and G. buchholzianum Engl.) domestication model to village-based farmers on and around Mount Cameroon. CARPE, Yaounde. 2000.

Marcio GC, Costa WO, Moore GA. An evaluation of factors affecting the efficiency of Agrobacterium-mediated transformation of Citrus paradisi (Macf.) and production of transgenic plants containing carotenoid biosynthetic genes. Plant Cell Reports. 2002; 21(4): 365-373.

Oliveira M, Marcio GC, Crislene V, Wagner CO. Growth regulators, culture media and antibiotics in the in vitro shoot regeneration from mature tissue of citrus cultivars Pesq. agropec. Bras, Brasília. 2010; 45(7): 654-660.

Liu G, Mai B. Adventitious shoot regeneration from in vitro cultured leaves of London plane tree (Platanus acerifolia Willd.). Plant Cell Reports . 2003. 21(7): 640-4.

Asseng ZA. Contribution à l’étude de la filière Gnetum sp (Gnetacées). Mémoire à l’Université de Yaoundé I. 2006.

Jiofack T, Fokunang C, Kemeuze V, Fongnzossie E, Tsabang N, Nkuinkeu R, Mapongmetsem PM, Nkongmeneck BA. Ethnobotany and phytopharmacopoea of the South-West ethnoecological region of Cameroon. J Med Plants Res. 2008; 2(8): 197–206.

Arvind Kumar GOYAL. Why MS media is most frequently used for plant tissue culture (internet). Bodoland University. 28th July 2016.

Philomena G, Jerold M. Low Cost Tissue Culture Technology for the Regeneration of Some Economically Important Plants for Developing Countries. International Journal of Agriculture, Environment & Biotechnology. 2013; 6: 703-711.

Remi NW, Chris O, Mathew MA, Mel OO.Good Agricultural Practices for African Indigenous Vegetables. Proceedings of a Technical Consultation Workshop Held in Arusha, Tanzania. 2009; 7‐ 8.

Murashige T, Skoog F. A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiol Plant. 1962; 15: 473–497.

Lloyd G, McCown B. Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip culture. Int. Plant Propagators’ Soc. Proc. 1980; 30.

Nitish k, ReddyIn M In vitro Plant Propagation: A Review. Journal of Forest Science. 2011; 27(2): 61-72.

Katalin MT, Nóra M, Alexandra H, László Z, Judit D. Phytotoxicity and Other Adverse Effects on the In vitro Shoot Cultures Caused by Virus Elimination Treatments: Reasons and Solutions. Plants (Basel). 2021; 10(4): 670.

Jinhui F, Yiyong Z, Cuiqing M, Xiaofeng C, Quan Z, Mingyou T, Bo Y, Ping X . The Surfactant Tween 80 Enhances Biodesulfurization. Appl Environ Microbiol. 2006; 72(11): 7390–7393.

Feng Z, Chong C, Lidong Cao, Fengmin Li, Fengpei Du, Qiliang H. Wetting Behavior and Maximum Retention of Aqueous Surfactant Solutions on Tea Leaves. Molecules. Jun 2019; 24(11): 2094.

George EF. Plant propagation by tissue culture. Part 1. The technology. Exegetics Ltd, Edington, England. 1993; 574.

Zok S, Sama AE, Nyochembeng L, Tambong JT, Ndzana X, Wutoh JG. Rapid multiplication of roots and tuber crops through tissue culture in Cameroon. In; Cahiers Agricultures. 1998; 7: 63-66.

Pei-Fen S, Chin-Jin L, Chih-Chien H, Spencer B, Sheng-Yang W, Kandan A, Sunney IC, Shih-Hsiung W, Feng-Ling Y, Wen-Ching H, Lie-Fen S, Ning-Sun Y. Dioscorea Phytocompounds Enhance Murine Splenocyte Proliferation Ex Vivo and Improve Regeneration of Bone Marrow Cells in Vivo. Evid Based Complement Alternat Med. 2011; 731308.

Wegayehu F, Firew M, Belayneh A. Optimization of explants surface sterilization condition for field grown peach (Prunus persica L. Batsch. Cv. Garnem) intended for in vitro culture. African Journal of Biotechnology. 2015; 14(8): 657-660.

Ninad VS, Shailendra SG. Cynodon Dactylon: A Systemic Review of Pharmacognosy, Phytochemistry and Pharmacology. International Journal Pharm Pharm Science. 2014; 6(8): 07-12.

Edwin FG, Michael AH, Geert-Jan De K. Plant Growth Regulators III: Gibberellins, Ethylene, Abscisic Acid, their Analogues and Inhibitors; Miscellaneous Compounds. George (eds.), Plant Propagation by Tissue Culture 3rd Edition. Plant Propagation by Tissue Culture. 2007; 227-281.

Cheesbrought M. Medical Laboratory Manual for Tropical countries. Vol. II: Microbiology. Tropical Health Technology / Butterworth - Heinemam. 1984; 479.

Thomas E. Biotechnology Applications of Plant Callus Culture. Engineering. 2018; 5(1): 50–59.