African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12484

Full Length Research Paper

Isolation and binary fusion of taro (Colocasia esculeneta (L) Schott.) protoplast: Towards developing somatic hybridization protocol as an alternative to sexual hybridization

Mohamed Chungwa
  • Mohamed Chungwa
  • Crops, Horticulture and Soils Department, Egerton University, P. O. Box 536-20115 Egerton, Njoro, Kenya.
  • Google Scholar
Robert Gesimba
  • Robert Gesimba
  • Crops, Horticulture and Soils Department, Egerton University, P. O. Box 536-20115 Egerton, Njoro, Kenya.
  • Google Scholar
Abwao S. Indieka
  • Abwao S. Indieka
  • Biochemistry and Molecular Biology Department, Egerton University, P. O. Box 536-20115 Egerton, Njoro, Kenya.
  • Google Scholar


  •  Received: 07 November 2023
  •  Accepted: 12 December 2023
  •  Published: 30 April 2024

Abstract

Collocasia esculenta (L) Schott. (Taro) genotypes cultivated in Kenya rarely produce flowers, thus improvement via pollination is hindered. Somatic hybridization is an attractive alternative to circumvent flower pollination constrain and hence explored in this study. The aim of this study was to optimize isolation and binary fusion of protoplast obtained from C. esculenta (Dasheen) and C. antiquorum Eddoe) genotypes. Protoplast was isolated from embryogenic calli and leaf tissues using cellulase R-10 (1.0% w/v) combined with pectinase R-10 (0.15% w/v). Fusion of leaf- and calli-derived protoplast was conducted using PEG 6000 (0-30% w/v), CaCl2 (0-0.15 M) and NaNO3 (0-4% w/v). Overall, 2 to 4 h enzyme incubation and PEG (10 or 20%) treatments for 10 to 20 min, were optimal for isolation of viable protoplast and fusion, respectively. The capacity of binary fusions to form cell colonies was higher when fusion was undertaken using PEG at either 10 or 20% after 10 and 20 min incubation. The study demonstrates that optimized protoplast fusion is a viable alternative for taro improvement that by passes flowering constrain.

 

Key words: Taro, protoplast, binary fusion, somatic hybridization, PEG 6000, heterokarya.