基于高通量测序的云南雪茄烟叶微生物群落结构及功能分析

Microbial community structure and function of Yunnan cigar tobacco leaves based on high-throughput sequencing

  • 摘要: 【目的】 探究云南不同产地及用途雪茄烟叶农业发酵后烟叶微生物群落结构和多样性,为雪茄烟叶功能微生物筛选及提高雪茄烟叶发酵品质提供参考。【方法】 以云南德宏、临沧和普洱3个产地不同用途(茄衣、茄芯、茄套)的雪茄烟叶为试验材料,基于高通量测序技术分析微生物群落结构,并分析预测微生物群落代谢功能。【结果】 按照97%相似性进行操作分类单元(OTU)聚类分析,共得到2393个OTUs,各个样品OTUs数在125~442,样品之间OTU数存在较大差异。同一产地不同用途烟叶样品的Chao1指数、ACE指数和Shannon指数有所差异,均表现为茄套烟叶高于茄衣和茄芯烟叶;同一用途不同产地烟叶样品比较,德宏和普洱2个产地烟叶样品的3个指数均高于临沧产地烟叶样品,具有更丰富的微生物种类。不同产地烟叶样品微生物以细菌为主,优势菌门主要为变形菌门(Proteobacteria)和厚壁菌门(Firmicutes)。不同用途烟叶优势菌属差异明显,3个产地茄衣烟叶中克雷伯杆菌属(Klebsiella)相对丰度占比较大,临沧和普洱茄芯烟叶中泛菌属(Pantoea)占主导地位,而3个产地茄套烟叶优势菌属种类更多元化且更丰富。KEEG和COG功能预测分析结果表明,9个烟叶样品微生物功能代谢途径和功能类型基本一致,但相对丰度存在较大差异。KEEG功能预测代谢通路主要有全局和概览地图、碳水化合物代谢、氨基酸代谢等;COG功能种类预测主要分布在未知功能、氨基酸运输和代谢、转录、无机离子运输和代谢、碳水化合物的运输和代谢等。【结论】 烟叶用途是影响云南雪茄烟叶微生物多样性的重要因素,烟叶产地的影响相对较小,茄套烟叶微生物丰富度高于茄衣和茄芯烟叶。

     

    Abstract: 【Objective】 To explore the microbial community structure and diversity of cigar tobacco leaves after agricultural fermentation in different producing areas and uses in Yunnan,which could provide reference for screening functional microorganisms of cigar tobacco leaves and improving the fermentation quality of cigar tobacco leaves. 【Method】 The microbial community structure was analyzed based on high-throughput sequencing technology,and the metabolic functions of the microbial community were analyzed and predicted using cigar tobacco leaves with different uses(cigar wrapper,filler and binder leaves)from 3 origins,namely Dehong,Lincang and Pu’er in Yunnan,as the test materials. 【Result】 Operation taxonomic unit(OTU)cluster analysis was performed according to 97% similarity,and a total of 2393 OTUs were obtained. The number of OTUs in each sample ranged from 125 to 442,and there was a large difference between samples. The Chao1 index,ACE index and Shannon index of tobacco leaf samples of different uses from the same origin were different,all showed binder > wrapper and filler leaves. Comparing tobacco leaf samples for the same use but from different origins,the 3 indexes of tobacco leaf samples from Dehong and Pu’er were higher than those of Lincang tobacco samples,with more abundant microbial species. The microorganisms of cigar tobacco leaves from different producing areas were mainly bacteria,and the dominant phylum were Proteobacteria and Firmicutes. There were great differences in the dominant genera of tobacco leaves with different uses,with Klebsiella occupying a larger proportion of the cigar wrapper leaves from 3 origins,Pantoea dominated in the filler leaves from Lincang and Pu’er;and the dominant genera of the binder leaves from 3 origins were more diverse and abundant. The results of KEEG and COG function prediction analysis showed that the microbial functional metabolic pathways and functional types of 9 tobacco samples were basically the same,but the relative abundance was greatly different. KEEG function prediction metabolic pathways were mainly global and overview maps,carbohydrate metabolism,amino acid metabolism,and the COG function type prediction was mainly distributed in the unknown function,amino acid transport and metabolism,transcription, inorganic ion transport and metabolism,carbohydrate transport and metabolism. 【Conclusion】 The use of tobacco leaves is an important factor affecting the microbial diversity of cigar tobacco leaves in Yunnan,while the influence of producing area is relatively small,and the microbial richness of binder leaves is higher than that of cigar wrapper and filler leaves.

     

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