不同饲料饲育的家蚕幼虫肠道细菌的多样性分析

doi:10.16380/j.kcxb.2019.01.007

摘要/Abstract

摘要：

【目的】为研究饲料对不同家蚕Bombyx mori品种肠道微生物菌群的影响。【方法】以筛选到的家蚕广食性品种GS和普通品种1015为研究对象，收集从收蚁开始分别饲育桑叶 (GS.m和C1015.m组)和人工饲料(GS.b组)至4龄盛时期的家蚕肠道样本，采用高通量测序的方法对其肠道微生物16S rDNA的V3-V4区进行测序分析，比较它们之间肠道微生物的差异。【结果】在门水平上，所测家蚕肠道样本的优势菌为厚壁菌门(Firmicutes)和变形杆菌门(Proteobacteria)；在科水平上，所测样本主要优势菌为明串珠菌科(Leuconostocaceae)、乳酸杆菌科(Lactobacillaceae)、肠杆菌科(Enterobacteriaceae)等；在属水平上，所测样本主要的优势菌为魏斯氏属Weissella、乳酸菌属Lactobacillus、布赫纳氏菌属Buchnera、甲基杆菌属Methylobacterium、叶瘤菌属Phyllobacterium、肠球菌属Enterococcus和脆弱拟杆菌属Bacteroides等。家蚕品种GS经桑叶和人工饲料饲育后，甲基杆菌属Methylobacterium、布赫纳氏菌属Buchnera等菌属仅在桑叶饲育的GS肠道内出现，而魏斯氏菌Weissella、短芽孢杆菌属Brevibacillus等菌属只在人工饲料饲育的GS肠道内出现。同是桑叶饲育的家蚕品种GS和1015，其肠道内相同的优势菌有叶瘤菌属Phyllobacterium、脆弱拟杆菌属Bacteroides、不动细菌属Acinetobacter等。相较于广食性蚕品种GS的肠道菌群，肠球菌属Enterococcus、草螺菌属Herbaspirillum、丝硫菌属Thiothrix等菌属仅在普通蚕品种1015肠道中被检测到。GS.b组家蚕肠道细菌的物种多样性低于GS.m和C1015.m。GS.m肠道中丰度差异显著性最高的菌群为变形菌门(Proteobacteria)，GS.b肠道中丰度差异显著性最高的菌群为杆菌纲(Bacilli)和乳杆菌目(Lactobacillales)，而C1015.m肠道中丰度差异显著性最高的菌群为粪肠球菌属Enterococcus和肠球菌科(Enterococcaceae)。【结论】经桑叶饲育的不同蚕品种(GS和1015)的肠道微生物比人工饲料饲育的家蚕肠道微生物更趋于一致；经桑叶饲育的广食性家蚕肠道微生物物种多样性较高于经人工饲料饲育的广食性家蚕。

关键词: 家蚕, 高通量测序, 16S rDNA, 肠道菌群, 物种多样性

Abstract: 【Aim】 This study aims to investigate the effects of feed on intestinal microflora of different silkworm (Bombyx mori) varieties. 【Methods】 The polyphagous silkworm strain GS and common silkworm strain 1015 were fed with mulberry leaves (groups GS.m and C1015.m) and artificial diet (group GS.b), respectively, and the intestinal samples of the 4th instar larvae of B. mori were collected. The V3-V4 region of the 16S rDNA of intestinal microorganisms was sequenced by using the high-throughput sequencing method, and the differences in intestinal microflora between the two strains were compared.【Results】 The results showed that the dominant bacteria in B. mori intestine at the phyla level were Firmicutes and Proteobacteria, those at the family level were Leuconostocaceae, Lactobacillaceae and Enterobacteriaceae, and those at the genus level were Weissella, Lactobacillus, Buchnera, Methylobacterium, Phyllobacterium, Enterococcus, Bacteroides, etc. After GS was fed with mulberry leaves and the artificial diet, Methylobacterium and Buchnera were present only in the intestine of GS fed with mulberry leaves, while Weissella and Brevibacillus appeared only in the intestine of GS fed with the artificial diet. The common dominant intestinal bacteria in silkworm strains GS and 1015 fed with mulberry leaves were Phyllobacterium, Bacteroides, Acinetobacter, etc. Compared with the intestinal microflora of the GS strain, Enterococcus, Herbaspirillum and Thiothrix were detected only in the strain 1015. The species diversity of intestinal bacteria in group GS.b was lower than those of groups GS.m and C1015.m. The intestinal bacteria whose abundance showed the most significant difference was Proteobacteria in GS.m, Bacilli and Lactobacillales in GS.b, and Enterococcus and Enterococcacea in C1015.m. 【Conclusion】 The intestinal microflora in different silkworm strains (GS and 1015) fed with mulberry leaves are more consistent as compared to that in silkworm strains fed with the artificial diet. The species diversity of intestinal microflora in the polyphagous silkworm fed with mulberry leaves is higher than that in the polyphagous silkworm fed with the artificial diet.

Key words: Bombyx mori, high-throughput sequencing, 16S rDNA, intestinal microflora, species diversity

郝长富, 李刚, 孙熙, 唐健, 钱荷英, 赵国栋, 邓祥元, 徐安英. 不同饲料饲育的家蚕幼虫肠道细菌的多样性分析[J]. 昆虫学报, 2019, 62(1): 61-72.

HAO Chang-Fu, LI Gang, SUN Xi, TANG Jian, QIAN He-Ying, ZHAO Guo-Dong, DENG Xiang-Yuan, XU An-Ying. Analysis of intestinal bacterial diversity in Bombyx mori larvae reared on different feeds[J]. Acta Entomologica Sinica, 2019, 62(1): 61-72.

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不同饲料饲育的家蚕幼虫肠道细菌的多样性分析

Analysis of intestinal bacterial diversity in Bombyx mori larvae reared on different feeds

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