Rapid identification of transgenic black pepper using loop-mediated isothermal amplification (LAMP) and real-time LAMP assays

A loop-mediated isothermal amplification (LAMP) and real-time LAMP based assays were developed for quick and sensitive detection of transgenic black pepper plants. Primers (six each) were designed based on the nucleotide sequence of two target regions [kanamycin and Cauliflower mosaic virus (CaMV) 3...

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Bibliographic Details
Published in:Journal of plant biochemistry and biotechnology 2015-10, Vol.24 (4), p.466-469
Main Authors: Sasi, Shina, Revathy, K. A., Bhat, A. I.
Format: Article
Language:English
Subjects:
Biomedical and Life Sciences
Cell Biology
Life Sciences
Plant Biochemistry
Protein Science
Receptors
Short Communication
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Summary:A loop-mediated isothermal amplification (LAMP) and real-time LAMP based assays were developed for quick and sensitive detection of transgenic black pepper plants. Primers (six each) were designed based on the nucleotide sequence of two target regions [kanamycin and Cauliflower mosaic virus (CaMV) 35S promoter] integrated into the genome of transgenic black pepper. Both assays successfully detected the transgenic plants and no cross-reaction was recorded with non-transgenic plants. The sensitivity of LAMP was up to 10 4 times that of conventional PCR while real-time LAMP was up to 10 3 times that of LAMP and 10 7 times to that of PCR. The addition of 6 mM magnesium sulphate and 0.4 M betaine with 1 h reaction time proved optimal for amplification through LAMP assay. The assays were validated by testing putative transformants of black pepper. The present study clearly established that LAMP and real-time LAMP assays can provide a rapid and simple approach for screening transgenic black pepper and other plants transformed by using the above target gene sequences.
ISSN:0971-7811
0974-1275
DOI:10.1007/s13562-015-0302-1