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

Aremu BR, Babalola OO - Int J Environ Res Public Health (2015)

Bottom Line: 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 Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Faculty of Agriculture, Science and Technology, North-West University, Private Bag X2046, Mmabatho 2735, South Africa. 23925523@nwu.ac.za.

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.

No MeSH data available.


Related in: MedlinePlus

Selective frequencies of the primer with respect to the macergens in the vegetables.
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ijerph-12-12356-f003: Selective frequencies of the primer with respect to the macergens in the vegetables.

Mentions: With the use of these designed primers, fourteen macergens were detected in sixteen vegetables out of twenty-six samples examined. Enterobacter sp., Lelliottia sp and Klebsiella sp. were detected by all the primer sets. The most abundant out of all the macergens detected is Citrobacter sp. detected by Primer Sets 1, 2, and 4 (Figure 3).


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

Aremu BR, Babalola OO - Int J Environ Res Public Health (2015)

Selective frequencies of the primer with respect to the macergens in the vegetables.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4626973&req=5

ijerph-12-12356-f003: Selective frequencies of the primer with respect to the macergens in the vegetables.
Mentions: With the use of these designed primers, fourteen macergens were detected in sixteen vegetables out of twenty-six samples examined. Enterobacter sp., Lelliottia sp and Klebsiella sp. were detected by all the primer sets. The most abundant out of all the macergens detected is Citrobacter sp. detected by Primer Sets 1, 2, and 4 (Figure 3).

Bottom Line: 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 Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Faculty of Agriculture, Science and Technology, North-West University, Private Bag X2046, Mmabatho 2735, South Africa. 23925523@nwu.ac.za.

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.

No MeSH data available.


Related in: MedlinePlus