基于高通量测序分析槲皮素对家蚕生长、产丝量及肠道菌群多样性的影响

Effects of quercetin on the growth,silk yield and gut microbiota diversity in silkworms based on high-throughput sequencing

  • 摘要: 【目的】基于高通量测序分析槲皮素对家蚕生长、产丝量及肠道菌群多样性的影响,探究槲皮素在家蚕肠道微生态调控中的作用,为功能性蚕丝的高质量生产提供理论依据。【方法】选取240头品种为9芙×7湘的家蚕分为3组,对照组(NC组)喂食普通桑叶,低剂量槲皮素组(LQR组)和高剂量槲皮素组(HQR组)分别喂食含有1.25%(w/w)和5.00%(w/w)槲皮素的桑叶,从五龄眠起(五龄第1 d)开始连续添食6 d,测定家蚕平均生长速率、存活率和蚕茧质量指标,利用高通量测序分析家蚕肠道菌群的组成。【结果】家蚕添食槲皮素后,平均生长速率减缓,存活率降低,HQR组的全茧量、茧层量和茧层率均显著低于NC组(P<0.05,下同)。添食槲皮素可增加家蚕肠道菌群的丰富度和多样性,HQR组和NC组的家蚕肠道菌群物种组成差异显著。在门分类水平上,各组的优势菌门均为变形菌门、蓝细菌门和厚壁菌门;与NC组相比,HQR组变形菌门的相对丰度显著降低。在属分类水平上,各组的优势菌属均为无色杆菌属、甲基杆菌属、norank_f__norank_o__Chloroplast、鞘氨醇单胞菌属和不动杆菌属;与NC组相比,HQR组无色杆菌属的相对丰度显著降低。KEGG信号通路分析结果显示,HQR组肠道菌群注释到全局和概述图及能量代谢通路的比例均显著高于NC组,而注释到氨基酸代谢和膜转运通路的比例均显著低于NC组;COG功能注释分析结果表明,HQR组注释到氨基酸转运和代谢、能量生成和转换及无机离子转运和代谢类别的比例均显著低于NC组,与KEGG信号通路分析结果相符,说明HQR组家蚕需要消耗更多的能量用于维持正常生长,同时蛋白质的生物合成受限制,不利于家蚕的生长和丝蛋白的合成。【结论】在桑叶中添加槲皮素会引起家蚕肠道菌群组成的变化,使变形菌门和无色杆菌属的相对丰度降低,导致家蚕能量代谢提高,氨基酸代谢、能量生成和转换及膜转运等方面能力下降,影响家蚕生长和产丝量。

     

    Abstract: 【Objective】This study employed high-throughput sequencing to evaluate the effects of quercetin on the growth,silk yield,and gut microbiota diversity of silkworms,to elucidate quercetin’s regulatory function on the silkworm gut microecology,thereby providing a theoretical foundation for the high-quality production of functional silk. 【Method】A total of 240 silkworms of the 9 Fu×7 Xiang breed were selected and divided into 3 groups:a control group (NC group)fed with ordinary mulberry leaves,a low-dose quercetin group(LQR group)fed with mulberry leaves containing 1.25%(w/w)quercetin,and a high-dose quercetin group(HQR group)fed with mulberry leaves containing 5.00%(w/w)quercetin. Beginning at the fifth instar(day 1 of the fifth instar),the silkworms were fed continuously for 6 d. The average growth rate,survival rate and cocoon quality indexes were measured,the silkworms’gut microbiota composition was analyzed via high-throughput sequencing.【Result】After quercetin was supplemented into the silkworms’diet, their average growth rate slowed down,and survival rate decreased. The cocoon mass,cocoon shell mass,and rate of cocoon shell in the HQR group were significantly lower than those in the NC group(P<0.05,the same below). The addition of quercetin enhanced the richness and diversity of the silkworm gut microbiota,with a significant difference in species composition observed between the HQR and NC groups. At the phylum level,the dominant bacterial phyla in all groups were Proteobacteria,Cyanobacteria,and Firmicutes. Compared with the NC group,the relative abundance of Proteobacteria in the HQR group was significantly reduced. At the genus level,the dominant bacterial genera in all groups were AchromobacterMethylobacteriumnorank_f__norank_o__ChloroplastSphingomonas and Acinetobacter. The relative abundance of Achromobacter in the HQR group was significantly reduced compared with the NC group. The KEGG signal pathway analysis results showed that the proportion of gut microbiota in the HQR group which annotated to global and overview maps as well as energy metabolism pathways was significantly higher than in the NC group. In contrast,the proportion that annotated to amino acid metabolism and membrane transport pathways was significantly lower than in the NC group. Similarly,COG functional annotation analysis revealed that the HQR group had a significantly lower proportion of annotations related to amino acid transport and metabolism,energy production and conversion,and inorganic ion transport and metabolism compared to the NC group. These findings aligned with the KEGG pathway analysis,suggesting that silkworms in the HQR group needed to expend more energy to sustain normal growth,while protein biosynthesis was hindered,negatively affecting both silkworm growth and silk protein synthesis.【Conclusion】The addition of quercetin to mulberry leaves alters the composition of the silkworm gut microbiota,reducing the relative abundance of Proteobacteria and Achromobacter. This shift leads to an increase in energy metabolism,but a reduction in the efficiency of amino acid metabolism,energy production and conversion,and membrane transport,ultimately impairing silkworm growth and reducing silk yield.

     

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