Department of Health Sciences, Universidade Estadual de Santa Cruz, Ilhéus 45662-900, BA, Brazil.
Department of Biological Science, Center of Biotechnology and Genetics, Universidade Estadual de Santa Cruz, Ilhéus 45662-900, BA, Brazil.
Viruses. 2024 Jul 31;16(8):1226. doi: 10.3390/v16081226.
plantations are of significant economic importance worldwide, primarily for chocolate production. During the harvest and processing of cocoa beans, they are subjected to fermentation either by microorganisms present in the environment (spontaneous fermentation) or the addition of starter cultures, with different strains directly contributing distinct flavor and color characteristics to the beans. In addition to fungi and bacteria, viruses are ubiquitous and can affect the quality of the fermentation process by infecting fermenting organisms, destabilizing microbial diversity, and consequently affecting fermentation quality. Therefore, in this study, we explored publicly available metatranscriptomic libraries of cocoa bean fermentation in Limon Province, Costa Rica, looking for viruses associated with fermenting microorganisms. Libraries were derived from the same sample at different time points: 7, 20, and 68 h of fermentation, corresponding to yeast- and lactic acid bacteria-driven phases. Using a comprehensive pipeline, we identified 68 viral sequences that could be assigned to 62 new viral species and 6 known viruses distributed among at least nine families, with particular abundance of elements from the phylum. Interestingly, 44 of these sequences were specifically associated with ssRNA phages () and mostly fungi-infecting viral families (, , and ). Of note, viruses from those families show a complex evolutionary relationship, transitioning from infecting bacteria to infecting fungi. We also identified 10 and 3 viruses classified within the and families, respectively. The quantification of the virus-derived RNAs shows a general pattern of decline, similar to the dynamic profile of some microorganism genera during the fermentation process. Unexpectedly, we identified narnavirus-related elements that showed similarity to segmented viral species. By exploring the molecular characteristics of these viral sequences and applying Hidden Markov Models, we were capable of associating these additional segments with a specific taxon. In summary, our study elucidates the complex virome associated with the microbial consortia engaged in cocoa bean fermentation that could contribute to organism/strain selection, altering metabolite production and, consequently, affecting the sensory characteristics of cocoa beans.
种植园在全球范围内具有重要的经济意义,主要用于巧克力生产。在可可豆的收获和加工过程中,它们要么通过环境中存在的微生物(自然发酵)进行发酵,要么添加起始培养物进行发酵,不同的菌株直接为豆子带来独特的风味和颜色特征。除了真菌和细菌外,病毒也无处不在,它们可以通过感染发酵生物、破坏微生物多样性,从而影响发酵质量,对发酵过程的质量产生影响。因此,在这项研究中,我们探索了哥斯达黎加利蒙省公开可用的可可豆发酵宏转录组文库,寻找与发酵微生物相关的病毒。这些文库来自于同一发酵样本的不同时间点:发酵 7、20 和 68 小时,对应于酵母和乳酸菌驱动的阶段。我们使用一个综合的流程,鉴定出了 68 条可能与发酵微生物相关的病毒序列,它们可以分配到 62 个新的病毒种和 6 个已知病毒中,这些病毒分布在至少 9 个科中,其中以 phylum 的元素特别丰富。有趣的是,这些序列中有 44 条与 ssRNA 噬菌体()和主要感染真菌的病毒家族(,和)特异性相关。值得注意的是,这些家族的病毒显示出复杂的进化关系,从感染细菌到感染真菌。我们还分别鉴定出了 10 条和 3 条归类于 和 家族的病毒。病毒衍生 RNA 的定量显示出与一些微生物属在发酵过程中的动态特征相似的下降趋势。出乎意料的是,我们鉴定出了与 narnavirus 相关的元素,它们与分段病毒物种具有相似性。通过探索这些病毒序列的分子特征,并应用隐马尔可夫模型,我们能够将这些额外的片段与特定的分类群相关联。总之,我们的研究阐明了与参与可可豆发酵的微生物联合体相关的复杂病毒组,这可能有助于选择生物体/菌株,改变代谢产物的产生,从而影响可可豆的感官特征。
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