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Multifragment melting analysis of yeast species isolated from spoiled fruits
Z Kesmen1, E Özbekar1, M E Büyükkiraz1
1Department of Food Engineering, Faculty of Engineering, Erciyes University, Kayseri, Turkey.
Aims:
In this study, we developed a novel approach, multifragment melting analysis (MFMA), based on the simultaneous analysis of the melting characteristics of multiple DNA fragments for the differentiation and identification of yeast species isolated from spoiled fruits.
Methods And Results:
In total, 183 yeast isolates recovered from spoiled fruit samples were differentiated and grouped using MFMA. Six different DNA fragments of the 26S rRNA gene showing a high interspecific heterogeneity were amplified, and the PCR products were individually subjected to MFMA. An excellent discrimination and classification were obtained when the melting characteristics of all target DNA fragments for each species were simultaneously assessed. Species-level identification was performed by sequence analysis of representative isolates from each group. In the fruit samples, Hanseniaspora uvarum (Kloeckera apiculata) was found to be the most frequently isolated species followed by members of the Pichia genus, namely P. kluyveri, P. fermentans and P. kudriavzevii (Issatchenkia orientalis).
Conclusions:
Multifragment melting analysis provided a rapid and reliable approach for discrimination and grouping of a large number of yeast isolates recovered from fruit samples prior to sequence analysis-based identification.
Significance And Impact Of The Study:
The MFMA has great potential for fast and accurate investigation of yeast communities associated with food spoilage to determine their sources and routes and to prevent contamination.
Insights
Multifragment melting analysis (MFMA) rapidly differentiates yeast species from spoiled fruits. This method accurately groups 183 isolates, identifying Hanseniaspora uvarum and Pichia species as most common.
Area of Science:
- Microbiology
- Molecular Biology
- Food Science
Background:
- Yeast identification is crucial for understanding food spoilage.
- Traditional methods can be time-consuming and labor-intensive.
- Rapid and accurate yeast differentiation is needed for food safety.
Purpose of the Study:
- To develop and validate a novel method, multifragment melting analysis (MFMA), for yeast species differentiation.
- To apply MFMA for identifying yeast isolates from spoiled fruit samples.
- To assess the efficiency of MFMA in grouping and identifying yeast communities.
Main Methods:
- Developed multifragment melting analysis (MFMA) using six 26S rRNA gene fragments.
- Amplified and analyzed melting characteristics of multiple DNA fragments simultaneously.
- Used sequence analysis for species-level identification of representative isolates.
Main Results:
- MFMA successfully differentiated and grouped 183 yeast isolates from spoiled fruits.
- Simultaneous analysis of multiple DNA fragments yielded excellent discrimination.
- Hanseniaspora uvarum and Pichia species (P. kluyveri, P. fermentans, P. kudriavzevii) were the most prevalent.
Conclusions:
- MFMA offers a rapid, reliable method for yeast isolate discrimination and grouping.
- MFMA is valuable for preliminary analysis before sequence-based identification.
- MFMA has potential for investigating yeast communities in food spoilage and preventing contamination.
