African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12269

Full Length Research Paper

Morphological and RAPD-marker characterization of Melia volkensii (Gürke) in vitro plants regenerated via direct and indirect somatic embryogenesis

Eliud Sagwa Mulanda*
  • Eliud Sagwa Mulanda*
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Yeremia Chuhila
  • Yeremia Chuhila
  • Department of Zoology and Wildlife Conservation, College of Natural and Applied Sciences, University of Dar es Salaam, P.O. Box 35064 Dar es Salaam, Tanzania.
  • Google Scholar
Ryan Musumba Awori
  • Ryan Musumba Awori
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Mark Ochieng Adero
  • Mark Ochieng Adero
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Nelson Onzere Amugune
  • Nelson Onzere Amugune
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Elijah Akunda
  • Elijah Akunda
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Jenesio Ikindu Kinyamario
  • Jenesio Ikindu Kinyamario
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Eliud Sagwa Mulanda*
  • Eliud Sagwa Mulanda*
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Yeremia Chuhila
  • Yeremia Chuhila
  • Department of Zoology and Wildlife Conservation, College of Natural and Applied Sciences, University of Dar es Salaam, P.O. Box 35064 Dar es Salaam, Tanzania.
  • Google Scholar
Ryan Musumba Awori
  • Ryan Musumba Awori
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Mark Ochieng Adero
  • Mark Ochieng Adero
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Nelson Onzere Amugune
  • Nelson Onzere Amugune
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Elijah Akunda
  • Elijah Akunda
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar
Jenesio Ikindu Kinyamario
  • Jenesio Ikindu Kinyamario
  • Plant Tissue Culture, Genetics and Molecular Biology Labs, School of Biological Sciences, University of Nairobi, P.O Box 30197-00100 Nairobi, Kenya.
  • Google Scholar


  •  Received: 10 December 2014
  •  Accepted: 30 March 2015
  •  Published: 15 April 2015

Abstract

Somaclonal variation induced in vitro during tissue culture can be a problem in clonal micropropagation of elite plants. This study investigated the extent of morphological and genetic similarity or dissimilarity between Melia volkensii in vitro plants (somatic seedlings) obtained via somatic embryogenesis and normal seedlings. Comparisons were made between in vitro plants regenerated directly from cotyledon explants, indirectly from zygotic embryos and normal seedlings of the same parent trees. Regeneration was achieved using half MS medium supplemented with 0.05 mg/l thidiazuron. Shoots were elongated in half MS with 0.1 mg/l BAP plus 0.01 mg/l IAA then rooted in half MS with 0.1 mg/l IBA and 0.1 mg/l NAA. Six morphometric and five meristic characters were used for the morphological characterization. PCR-RAPD markers were used for assessment of genetic similarity or distance. Multivariate analysis using principal coordinates, cluster analysis, analysis of similarities (Anosim) and similarity percentages analysis (SIMPER) revealed significant dissimilarities (p< 0.0001) in morphometric and meristic characters between the in vitro plants and normal seedlings. However, significant similarity (p<0.01) was observed in the RAPD-genic characters of the in vitro plants and normal seedlings. Out of six morphometric traits, taproot length, internode length and shoot height were the most important sources of dissimilarity, cumulatively accounting for 72.37% of overall morphometric dissimilarity. Number of lateral roots was the single most important source of meristic dissimilarity, with 77.02% contribution. Plants regenerated directly from cotyledons were more similar to the normal seedlings in morphological and RAPD-marker characters than those regenerated indirectly from zygotic embryos. This study paves the way for identification of trait-specific RAPD markers for further characterization through sequence-characterized amplified regions (SCARs). 
 
Key words: Morphometric, meristic, PCR-RAPD, Melia volkensii, tissue culture, somaclonal variation.

Abbreviation

SCARs, Sequence-characterized amplified regions; RAPD, random amplified polymorphic DNA; SSR, simple sequence repeats; PCR-RFLP, polymerase chain reaction-restriction fragment length polymorphism; AFLP, amplified fragment length polymorphism; PCOORDA, principal coordinates analysis; Anosim, analysis of similarity; SIMPER, similarity percentage analysis.