Identification of Lactic Acid Bacterial Profile Effective on the Quality of Silages Prepared from Buckwheat (Fagopyrum esculentum Moench) by DNA Fingerprinting Techniques

Abstract

In this study, it was aimed to obtain data about dominant strains and to determine lactic acid bacteria (LAB) profile in the silage samples prepared from buckwheat (Fagopyrum esculentum Moench), grown in fields of the Faculty of Agriculture of Aydın Adnan Menderes University in Turkey's Aegean region, by using PCR-DNA-fingerprinting technique through M13 and (GTG)5 repeating sequence primers. In the study, 15 different silage samples prepared by adding 2 different concentrations (1.5x105-1.5x106 cfu g-1) of inoculant and 2 different concentrations (4-6 L t-1) of formic acid to buckwheat samples with different dry matter (DM) contents were used. First, DM, pH and Fleig points of 15 different silage samples, then total LAB numbers and the variety of LABs at genus level of these samples were determined. 50 isolates which were detected to be LAB from silage samples were obtained and Polymerase Chain Reaction (PCR) fingerprint profiles of these isolates were obtained using primers (GTG) 5 and M13 to detect LAB variety at species level. For the 50 LAB, a total of 52 loci with the M13 primer and 39 loci with the (GTG)5 primer were produced. The markers [M13 and (GTG)5] produced many different fragment profiles. According to the results of cluster and separation analysis, it was determined that both primers [M13 and (GTG)5] should be used for molecular characterization of LAB, isolated from silage samples. When samples with different DM ratio (20%, 30%, 40%) were examined in terms of microbial diversity and quality, 30% DM content came into prominence. The rise of the number of Enterococcus in samples with 30% and 40% DM content and the increase in silage quality in parallel shows that Enterococcus number can be effective on silage quality. Two of the isolates obtained in this study have the potential to become a silage starter. In the light of these data, it can be said that buckwheat silage containing 30% DM will be a quality silage with high LAB diversity and an alternative roughage.

Keywords

• silage
• buckwheat
• dna-fingerprinting
• enterococcus
• formic acid