PCR-based Specific Detection of Bacillus in Liquid Organic Fertilizer

Iman Hidayat(1*), Nur Laili(2), Dwi Agustiyani(3), Sarjiya Antonius(4)

(1) Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences-LIPI, Jalan Raya Jakarta-Bogor Km 46, Cibinong 16911, Indonesia
(2) Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences-LIPI, Jalan Raya Jakarta-Bogor Km 46, Cibinong 16911, Indonesia
(3) Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences-LIPI, Jalan Raya Jakarta-Bogor Km 46, Cibinong 16911, Indonesia
(4) Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences-LIPI, Jalan Raya Jakarta-Bogor Km 46, Cibinong 16911, Indonesia
(*) Corresponding Author

Abstract


Rapid molecular PCR-based detection method for Bacillus species used in the production of Beyonic® liquid organic fertilizer was carried out based on nucleotide sequence data from the 16S rRNA gene. The method involved sequencing the 16S rRNA gene of several Bacillus species and identifying around 16-22 specific nucleotide bases from 5' and 3' ends in the Bacillus 16S rRNA gene sequences. One specific primer pair for Bacillus detection was determined as follow: 5' - CAT AAG ACT GGG ATA ACT CCG GG - 3' (forward) from positions of 85-107 bp, and 5’ - CCA GGC GGA GTG CTT AAT GC - 3’ (reverse) from positions of 836-854 bp. PCR assay and gel electrophoresis analysis showed that the primer pair was specific to the genus Bacillus.


Keywords


bacteria, detection, molecular marker, organic fertilizer, 16S rRNA

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References


Genaid. 2017. Instruction manual - Presto™ Mini gDNA Bacteria Kit. Geneaid Biotech Ltd., Taiwan.

Hidayat I. 2016. Pengembangan Metoda Deteksi Bakteri Agens POH Startmik-Beyonic® dan Uji Simbosis DSE Terhadap Cabai dan Chinese Cabbage. Laporan Teknik. Pusat Penelitian Biologi LIPI. (In Indonesian language)

Promega. 2016. GoTaq® Green Master Mix. Promega Corporation




DOI: https://doi.org/10.37604/jmsb.v1i1.21

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