Research Article
Determination of Cyanogenic Compounds Content in Transgenic Acyanogenic Kenyan Cassava (Manihot esculenta Crantz) Genotypes: Linking Molecular Analysis to Biochemical Analysis
Ngugi M Piero1*, Murugi N Joan2, Oduor O Richard1, Mgutu A Jalemba1, Ombori R Omwoyo3 and Cheruiyot R Chelule3
1Department of Biochemistry and Biotechnology, Kenyatta University, Nairobi, Kenya
2Department of Environmental and Population Health, Kenyatta University, Nairobi, Kenya
3Department of Plant Sciences, Kenyatta University, Nairobi, Kenya
- *Corresponding Author:
- Mathew Piero Ngugi
School of Pure and Applied Sciences
Department of Biochemistry and Biotechnology
Kenyatta University, P.O Box 43844-00100
Nairobi, Kenya
Tel: +254 710 349 485/+254 732 264 557
E-mail: piero.mathew@ku.ac.ke/matpiero@gmail.com
Received date: July 17, 2015; Accepted date: August 05, 2015; Published date: August 12, 2015
Citation: Piero NM, Joan MN, Richard OO, Jalemba MA, Omwoyo OR, et al. (2015) Determination of Cyanogenic Compounds Content in Transgenic Acyanogenic Kenyan Cassava (Manihot esculenta Crantz) Genotypes: Linking Molecular Analysis to Biochemical Analysis. J Anal Bioanal Tech 6:264. doi:10.4172/2155-9872.1000264
Copyright: © 2015 Piero NM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Being the fourth most important crop in the developing countries surpassed only by maize, rice and sugarcane as a source of calories, cassava (Manihot esculenta Crantz) is doubtlessly a famine reserve crop due to its drought tolerance, ability to grow on infertile soils and its ability to recover from disease and pest attacks. However, this important tuber crop has a fair share of demerits, among which is the fact that all parts of the plant contain toxic levels of cyanogenic glycosides, which have to be removed by laborious processing before cassava can be safely consumed. Conventional methods for removal of cyanogenic glycosides in cassava have seldom been successful for decades. Genetic engineering holds the key to overcoming majority of these limitations in order to produce cassava plants in which desirable traits are optimized and undesirable traits downregulated. The objective of this study was to determine the levels of cyanogenic compounds in three Kenyan cassava genotypes along with an exotic model cultivar in which cyanogenic glycosides had been downregulated via RNA interference approach in an earlier study by the authors. Cassava roots from transgenic and wild type genotypes were harvested, peeled, cut to pieces and washed three times with cold water, following cyanogenic compounds were extraction by homogenization in acid extraction medium. The supernatant obtained after centrifugation of the homogenate was analyzed for cyanogenic compounds content by spectrophotometric procedures. From this study, transgenic cassava lines with cyanide content three folds less than the cyanide content in the wild type relatives were produced. This confirmed RNAimediated downregulation of expression of cytochrome P450 genes responsible for biosynthesis of cyano-glycosides previously undertaken by the authors in an earlier study.