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Int. J. Environ. Res. Public Health 2015, 12(10), 12356-12370; doi:10.3390/ijerph121012356

Construction of Specific Primers for Rapid Detection of South African Exportable Vegetable Macergens

1
Department of Biological Sciences, Faculty of Agriculture, Science and Technology, North-West University, Private Bag X2046, Mmabatho 2735, South Africa
2
Food Security and Safety Niche Area, Faculty of Agriculture, Science and Technology, North-West University, Private Bag X2046, Mmabatho 2735, South Africa
*
Author to whom correspondence should be addressed.
Academic Editor: Paul B. Tchounwou
Received: 24 August 2015 / Revised: 19 September 2015 / Accepted: 23 September 2015 / Published: 30 September 2015
View Full-Text   |   Download PDF [843 KB, uploaded 30 September 2015]   |  

Abstract

Macergens are bacteria causing great damages to the parenchymatous tissues of vegetable both on the field and in transit. To effectively and rapidly investigate the diversity and distribution of these macergens, four specific primers were designed by retrieving 16S rDNA sequences of pectolytic bacteria from GenBank through the National Center for Biotechnology Information (NCBI). These were aligned using ClusterW via BioEdit and primers were designed using Primer3Plus platform. The size and primer location of each species and PCR product size were accurately defined. For specificity enhancement, DNA template of known macergens (Pectobacterium chrysanthermi) and fresh healthy vegetable were used. These primers yielded expected size of approximately 1100 bp product only when tested with known macergens and no amplicon with fresh healthy vegetable was detected. Rapid detection of macergens in rotten vegetable samples was then carried out using these primers. Nucleotide sequences of macergens identified were deposited into the GenBank and were assigned accession numbers. Hence, with these specific primers, macergens can be identified with minimal quantities of the vegetable tissues using molecular techniques, for future use of the quarantine section of the Agricultural Department of the country for quick and rapid detection of macergens before exportation. View Full-Text
Keywords: ClusterW; macergens; pectolytic; rapid detection; specific primer; vegetable ClusterW; macergens; pectolytic; rapid detection; specific primer; vegetable
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Aremu, B.R.; Babalola, O.O. Construction of Specific Primers for Rapid Detection of South African Exportable Vegetable Macergens. Int. J. Environ. Res. Public Health 2015, 12, 12356-12370.

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Int. J. Environ. Res. Public Health EISSN 1660-4601 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
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