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Multiplex PCR for the detection of African cassava mosaic virus and East African cassava mosaic Cameroon virus in cassava
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A multiplex PCR was developed for simultaneous detection of African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV) in cassava affected with cassava mosaic disease (CMD). One set of three primers consisting of an upstream primer common for both viruses and two down stream virus-specific primers were designed for simultaneous amplification of 368 base pair (bp) and 650 bp DNA fragments specific to the replicase gene of ACMV and EACMCV, respectively. Similarly, a second set of three primerswere designed for simultaneous amplification of 540 bp and 655 bp fragments specific to the coat protein gene of EACMCV and ACMV, respectively. Primers that can amplify a 171 bp fragment of the large subunit of ribulose bisphosphate carboxylase oxygenase Lwere included as an internal control in these assays to determine the reliability of multiplex PCR. A simplified, cost-effective and rapid sample preparation method was adapted in place of the conventional plant DNA extraction procedure for multiplex PCR detection of ACMV and EACMCV. The method was validated using CMD-infected cassava samples obtained from farmers’ fields in Nigeria. The multiplex PCR is useful for reliable assessment of the prevalence of CMBs in epidemiological studies and for crop improvement and phytosanitary programs in African countries.
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Permanent link to this itemhttps://hdl.handle.net/20.500.12478/2894
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Alternate hosts of African cassava mosaic virus and East African cassava mosaic Cameroon virus in Nigeria Alabi, O.J.; Ogbe, F.O.; Bandyopadhyay, Ranajit; Kumar, P.L.; Dixon, Alfred G.O.; Hughes, Jacqueline d'Arros; Naidu, R.A. (2008)Cassava mosaic disease (CMD) caused by African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV) is the major constraint to cassava production in Nigeria. Sequences of the DNA-A component of ACMV and EACMCV isolates from leguminous plant species (Senna occidentalis, Leucana leucocephala and Glycine max), castor oil plant (Ricinus communis), a weed host (Combretum confertum) and a wild species of cassava (Manihot glaziovii) were determined. All ACMV isolates from ...
The occurrence of African cassava mosaic virus and East African cassava mosaic Cameroon virus in natural hosts other than cassava in Nigeria Alabi, O.J.; Ogbe, F.O.; Bandyopadhyay, Ranajit; Dixon, Alfred G.O.; Hughes, J.; Naidu, R.A. (2007)Cassava mosaic disease (CMD) in Africa is caused by six distinct cassava mosaic viruses (family eminiviridae, genus Begomovirus). Among them, African cassava mosaic virus (ACMV) and East African cassava mosaicCameroon virus (EACMCV) have been reported in CMD-infected cassava plants in Nigeria. Based on PCR tests using virus-specific primers, a leguminous plant Senna occidentalis and the weed Combreturn confertumwere previously identified as natural hosts for ACMV and EACMCV and castor oil plant ...
First report of the occurrence of African cassava mosaic virus in a mosaic disease of soybean in Nigeria Mgbechi-Ezeri, J.U.; Alabi, O.J.; Naidu, R.A.; Kumar, P.L. (2008)African cassava mosaic virus (ACMV; genus Begomovirus, family Geminiviridae) is one of six viruses documented in cassava (Manihot esculenta Crantz.) plants showing cassava mosaic disease in sub-Saharan Africa (SSA). In addition to cassava, the natural host range of ACMV includes a few wild Manihot species, Jatropha multifida, and Ricinus communis L. in Euphorbiaceae, and Hewittia sublobata in Convolvulaceae. The experimental host range of ACMV includes Nicotiana sp. and Datura sp. in the Solanaceae ...