Brachylaena huillensis (Silver Oak) is a multipurpose timber tree species in the family of Asteraceae. There has been a very high demand for B. huillensis wood and its products leading to overexploitation. B. huillensis regenerates through seedlings . However, it produces seeds with poor germination . Seeds are also difficult to be collect ed because of small size . Many are eaten by insects and currently there is a lack of seed bank . The facts that have hindered and rendered the natural regeneration of the tree species were uncertain. The present investigation was carried out to develop sanitation protocol of B. huillensis using leaves as explants. Juvenile leaves from the tips of B. huillensis naturally growing seedlings were collected from Bombo West Forest Reserve in Tanzania. The leaves were washed and immersed in NaOCL containing various concentrations levels and two drops of tween 20. There was significant difference between the concentrations levels employed . However , the best results were obtained when leaf explants w ere immersed in 1.5% v/v NaOCL for ten minutes and later in ethanol for ten seconds and cultured on woody plant media medium containing antifungal (cefotaxime). Genuinely, the protocol is vital and so opens up the way for other subsequent stages for in vitro propagation of B. huillensis .
WCMC, “Provision of Data on Rare and Threatened Tropical Timber Species,” Chapman and Hall Ltd., London, 1991, 58 Pages.
S. K. Chonge, “Study of Economic Aspects of the Wood Carving Industry in Kenya: Implications for Policy Development to Make the Industry More Sustainable,” Thesis for Award of Msc Degree at University of Natal, Natal, 2002, 55 pages. Unpublished.
C. K. Ruffo and S. M. Maliondo, “Forest Plant Genetic Resources in Tanzania,” Proceedings of the 1st National Workshop, Arusha, 16-20 January 1990.
IUCN, “IUCN Red List of Threatened Species,” 2008. http://www.iucnredlist.org
L. P. Mbuya, H. P. Msanga C. K. Ruffo A. Birniel and B. Tengnas, “Useful Trees and Shrubs for Tanzania: Identification, Propagation and Management for Agricultural and Pastoral Communities,” Regional Soil Conservation Unit, Nairobi, 1994, 542 Pages.
B. Cunningham, “Kenya’s Carvings, the Ecological Footprint of the Wooden Rhino Africa,” Wildlife and Environment, Vol. 6, No. 2, 1998, pp. 43-50.
J. M. Bryce and A. W. Chihongo, “The Commercial Timbers of Tanzania,” KAD Publishers, Dar es Salaam, 1999, 293 Pages.
N. T. Marshall and M, Jenkins, “Hard Times for Hardwood: Indigenous Timber and the Timber Trade in Kenya Traffic International,” London, 1994, 53 Pages.
H. N. Daud, J. Shashita and R. Mohamed, “An Improved Surface Sterilization Technique for Introducing Leaf, Nodal and Seed Explants of Aquilaria malaccensis from Field Sources into Tissue Culture,” Asia-Pacific Journal of Molecular Biology and Biotechnology, Vol. 20, No. 2, 2012, pp. 55-58
S. Seeni and P. G. Latha, “In Vitro Multiplication and Ecorehabilitation of the Endangered Blue Vanda,” Plant Cell, Tissue and Organ Culture, Vol. 61, 2000, pp. 1-8. http://dx.doi.org/10.1023/A:1006444614657
H. S. Chawla, “Plant Biotechnology: Laboratory Manual for Plant Biotechnology,” Oxford & IBH Publishing Co. Pvt Ltd., New Delhi, 2003.
A. Badoni and J. S. Chauhan, “In Vitro Sterilization Protocol for Micropropagation of Solanum tuberosum cv ‘Kufri Himalini’,” Academia Arena, Vol. 2, No. 4, 2010, pp. 24-27.
G. R. T. Rout, S. Samantaray and P. Das, “In Vitro Manipulation and Propagation of Medicinal Plants,” Biotechnology Advances, Vol. 18, No. 2, 2000, pp. 91-120. http://dx.doi.org/10.1016/S0734-9750(99)00026-9
J. C. Lovett and T. Pocs, “Assessment of the Condition of the Catchment Forest Reserves: A Botanical Appraisal,” Government Printers, Dar es Salaam, 1993, 300 Pages.
B. H. McCown, “Recalcitrance of Woody and Herbaceous Perennial Plants: Dealing with Genetic Predetermination,” In Vitro Cellular and Developmental Biology Plant, Vol. 36, No. 3, 2000, pp. 149-154. http://dx.doi.org/10.1007/s11627-000-0030-6
R. G. D. Steel and J. H. Torrie, “Principles and Procedures of Statistics,” 2nd Edition, McGraw Hill Book Co. Inc., New York, 1980, pp. 232-249.
I. B. Omamor, A. O. Asemota, C. R. Eke and E. I. Eliashi, “Fungal Contamination of the Oil Palm Tissue Culture in Nigerian Institute for Oil Palm Research (NIFOR),” African Journal of Agriculture Research, Vol. 2, No. 10, 2007, pp. 534-537.
O. I. Odutayo, N. A. Amusa, O. O. Okutade and Y. R. Ogunsanwo, “Sources of Microbial Contamination in Tissue Culture Laboratories in Southwestern Nigeria African,” Journal of Agricultural Research, Vol. 2, No. 3, 2007, pp. 67-72.
S. K. Webster, J.A. Seymour, S.A. Mitchell and M. H. Ahmad, “A Novel Surface Sterilization Method for Reducing Microbial Contamination of Field Grown Medicinal Explants Intended for In Vitro Culture,” Biotechnology Centre, Kingston, 2003.
P. J. Mathias, P. G. Alderson and R. R. B. Leakey, “Bacterial Contamination in Tropical Hardwood Cultures,” Acta Horticulture, Vol. 212, 1987, pp. 43-48
N. Brassard, C. Richer, D. Tousignant and J. A. Rioux, “Multiplication végé Tative de L’acer Saccharum: Contribution,” Micropropagation, Vol. 33, 2003, pp. 682-690.
H. R. Kerns and M. M. Meyer, “Tissue Culture Propagation of Acer X Freemanii Using Thidiazuron to Stimulate Shoot Tip Proliferation,” Hortisciences, Vol. 21, No. 5, 1986, pp. 1209-1210.
M. G. Padilla and C. L. Encina, “Micropropagation of Adult Cherimoya (Annona cherimola Mill ) cv Fino de Jete,” In Vitro Cellular & Developmental Biology—Plant, Vol. 40, No. 2, 2004, pp. 210-214. http://dx.doi.org/10.1079/IVP2003521
J. E. Preece, “Practical Regulation of Woody Plant Growth and Development Using Biotechnology,” Acta Horticulturae, Vol. 300, 1991, pp. 23-33.
M. Kane, “Bacterial and Fungal Indexing of Tissue Cultures,” 2003
O. B. Oyebanji, O. Nweke, O. Odebunmi, N. B. Galadima, M. S. Idris, U. N. Nnodi, A. S. Afolabi and G. H. Ogbadu, “Simple, Effective and Economical Explant-Surface Sterilization Protocol for Cowpea, Rice and sorghum seeds,” African Journal of Biotechnology, Vol. 8, No. 20, 2009, pp. 5395-5399.
N. Srivastava, B. Kamal V. Sharma, Y. K. Negi, A. K. Dobriyal, S. Gupta and V. S. Jadon, “Standardization of Sterilization Protocol for Micropropagation of Aconitum Heterophyllum—An Endangered Medicinal Herb,” Academia Arena, Vol. 2, No. 6, 2010, 37-42.
M. S. Greenwood, “Rejuvenation in Forest Trees,” Journal of Plant Growth Regulation, Vol. 6, 1987, pp. 1-12. http://dx.doi.org/10.1007/BF00043947
H. Colgecen, U. Koca and G. Toker, “Influence of Different Sterilization Methods on Callus Initiation and Production of Pigmented Callus in Arnebia Densifl ora Ledeb,” Turkish Journal of Biology, Vol. 35, No. 4, 2011, pp. 513-520.
A. N. Ahmed, “In Vitro Production of Somatic Embryos from Nucellus of Mango (Mangifera indica L.),” Life Science Journal, Vol. 10, No. 2, 2013, pp. 1164-1174.
A. S. Oluwaseun and A. B. Erhinmeyoma of Callus and Somatic Embryogenesis from Cotyledon Explants of Parkia biglobosa (Jacq ) Benth,” African Journal of Biotechnology, Vol. 4, No. 1, 2005, pp. 68-71.
T. C. Siang, S. T. Soong, A. Yien and T. Su, “Plant Regeneration Studies of Jatropha curcas Using Induced Embryogenic Callus from Cotyledon Explants,” African Journal of Biotechnology, Vol. 11, No. 31, 2012, pp 8022-8031
R. H. Smith, “Plant Tissue Culture Techniques and Experiments,” 2nd Edition, Academic Press, San Diego, 2000, 231 Pages.
S. S. Bhojwani and M. K. Rozdan, “Plant Tissue Cultue: Theory and Practice: Development in Crop Science,” Elsevier, Amsterdam, 1983.
M. N. Amin and V. S. Jaiswal, “In Vitro Response of Apical Bud Explants from Immature Trees of Jackfruit (Artocarpus heterophyllus),” Plant Cell, Tissue and Organ Culture, Vol. 33, 1993, pp. 59-65. http://dx.doi.org/10.1007/BF01997599
V. S. Jaiswal and M. N. Amin, “In Vitro Shoot Proliferation and Plantlet Formation from Somatic Tissues of Guava (Psidium guajava L.),” 6th International Congress of Plant Tissue and Cell Culture, University of Minnesota, Minneapolis (Abstract), 1986, p. 279.
B. M. Reed and P. Tanprasert, “Detection and Control of Bacterial Contaminants of Plant Tissue Cultures. A Review of Recent Literature,” Plant Tissue Culture and Biotechnology, Vol. 1, No. 3, 1995, pp. 137-142.
S. M. Woo and H. Y. Wetzstein, “Morphological and Histological Evaluations of in Vitro Regeneration in Elliottia racemosa Leaf Explants Induced on Media with Thidiazuron,” Journal of the American Society for Horticultural Science, 2008 (in Press).
R. Mohamed, P. L. Jong and M. S. Zali, “Fungal Diversity in Wounded Stems of Aquilaria Mala,” Fungal Diversity, Vol. 43, No. 1, 2010, pp. 67-74. http://dx.doi.org/10.1007/s13225-010-0039-z
K. Welsh, K. C. Sink and H. Davidson, “Progress on in Vitro Propagation of Red Maple,” Combined Proceedings—International Plant Propagators’ Society, Vol. 29, 1979, pp. 382-386.