昆虫学报 ›› 2022, Vol. 65 ›› Issue (2): 176-186.doi: 10.16380/j.kcxb.2022.02.006

• 研究论文 • 上一篇    下一篇

更换饲料对家蚕幼虫肠道细菌组成的影响及关键物种与产茧性能的相关性

张雨丽1,#, 曾珠2,#, 刘艳伟1, 陆俣伽1, 韦伟1鲁成2, 崔为正3, 闭立辉1, 王平阳1, 张桂征1,*   

  1. (1. 广西蚕业科学研究院, 南宁 530007; 2. 西南大学蚕桑纺织与生物质科学学院, 重庆 400715; 3. 山东农业大学林学院, 山东泰安 250100)
  • 出版日期:2022-02-20 发布日期:2022-03-10

Effects of feed change on intestinal bacterial composition in Bombyx mori larvae and the correlation between the key species and cocoon production performance

ZHANG Yu-Li1,#, ZENG Zhu2,#, LIU Yan-Wei1, LU Yu-Jia1, WEI Wei1, LU Cheng2, CUI Wei-Zheng3, BI Li-Hui1, WANG Ping-Yang1, ZHANG Gui-Zheng1,*    

  1. (1. Guangxi Academy of Sericultural Sciences, Nanning 530007, China; 2. College of Sericulture, Textile and Biomass Sciences, Southwest University, Chongqing 400715, China; 3. College of Forestry, Shandong Agricultural University, Tai′an, Shandong 250100, China)
  • Online:2022-02-20 Published:2022-03-10

摘要: 【目的】本研究探讨使用人工饲料部分替代桑叶饲育后家蚕Bombyx mori幼虫肠道细菌群落的变化情况,并分析人工饲料替代桑叶的不同饲育模式下肠道关键细菌与蛹重和茧层量的相关性,为人工饲料部分替代法饲育家蚕的实际应用提供理论依据。【方法】采用全龄桑叶育(Mul1-5)、1-2龄人工饲料育+3-5龄桑叶育(Art1-2)、1-3龄人工饲料育+4-5龄桑叶育(Art1-3)、1-4龄人工饲料育+5龄桑叶育(Art1-4)和全龄人工饲料育(Art1-5) 5种饲养模式饲养家蚕幼虫,统计5种饲养模式下家蚕的全茧量、茧层量及蛹重;收集不同饲养模式下家蚕5龄幼虫肠道样品,通过高通量测序技术分析其肠道菌群的组成和多样性差异,采用Spearman相关性热图分析肠道细菌与蛹重和茧层量的相关性。【结果】人工饲料与桑叶不同搭配的饲育模式对家蚕茧质有显著的影响,Mul1-5和Art1-5的全茧量最高,二者无显著性差异,Art1-2其次; Mul1-5茧层量最高,Art1-2其次,Art1-5茧层量最低。在不同饲育模式下,家蚕肠道细菌多样性和组成显著不同:在门水平上,Art1-2和Mul1-5的细菌组成最相近,优势菌主要是变形菌门(Proteobacteria)和厚壁菌门(Firmicutes),而Art1-3, Art1-4和Art1-5的细菌组成最相近,变形菌门丰度随着人工饲料饲养时间的增加逐渐增加,厚壁菌门的丰度则呈现相反的趋势;在属水平上,家蚕肠道优势菌属有假单胞菌属Pseudomonas、肠球菌属Enterococcus、鞘脂单胞菌科未分类菌属Sphingomonadaceae unclassified及罗尔斯通菌属Ralstonia等。不同饲育模式下家蚕肠道菌属丰度差异最大的是肠球菌属,Art1-2组中肠球菌属丰度最高(40.9%),随着人工饲料饲养时间的延长,丰度逐渐降低,在Art1-5中仅为0.02%。家蚕肠道菌群大部分关键物种均与茧质呈一定的相关性:柯克斯体属Coxiella与蛹重呈显著正相关,与茧层量呈显著负相关;葡萄球菌属Staphylococcus与蛹重呈显著负相关;肠球菌属与茧层量呈显著正相关。【结论】饲育模式Art1-2最接近于Mul1-5,可为人工饲料部分替代桑叶饲育法饲育家蚕提供参考。人工饲料部分替代桑叶后家蚕肠道菌群结构发生了显著变化,肠球菌属丰度的显著降低及假单胞菌属的显著增加可能与全龄人工饲料养家蚕体质弱有关;一些关键细菌与家蚕蛹重和茧层量存在显著相关性,其影响机制需要进一步研究。

关键词: 家蚕, 人工饲料, 桑叶, 肠道细菌, 产茧性能

Abstract: 【Aim】This study aims to analyze the changes of intestinal bacterial communities in Bombyx mori larvae and the correlation between the key intestinal bacteria and the pupal weight and cocoon layer weight under different rearing patterns of artificial diet instead of mulberry leaves, so as to provide theoretical basis for the practical application of rearing B. mori by the method of partial artificial diet replacement of mulberry leaves.【Methods】B. mori larvae were reared under five different patterns including reared on mulberry leaves for all instars (Mul1-5), reared on artificial diet for the 1st-2nd instars and mulberry leaves for the 3rd-5th instars (Art1-2), reared on artificial diet for the 1st-3rd instars and mulberry leaves for the 4th-5th instars (Art1-3), reared on artificial diet for the 1st-4th instars and mulberry leaves for the 5th instar (Art1-4), and reared on artificial diet for all instars (Art1-5), and the whole cocoon weight, cocoon layer weight and pupal weight were calculated. The intestinal samples of the 5th instar larvae of B. mori under the five different rearing patterns were collected and their differences in composition and diversity of intestinal bacteria were analyzed by the high-throughput sequencing method. The correlation between the intestinal bacteria and the pupal weight and cocoon layer weight was analyzed by the Spearman correlation heatmap analysis. 【Results】The rearing patterns combining artificial diet with mulberry leaves had significant effects on cocoon quality of B. mori. The whole cocoon weight of Mull1-5 and Art1-5 was the highest, showing no significant difference, followed by that of Art1-2. The cocoon layer weight of Mul1-5 was the highest, followed by that of Art1-2, and that of Art1-5 was the lowest. The diversity and composition of intestinal bacteria in B. mori under the five rearing patterns were significantly different. At the phylum level, the bacterial composition of Art1-2 and Mul1-5 were the most similar, with the dominant bacteria of Proteobacteria and Firmicutes, while the bacterial composition of Art1-3, Art1-4 and Art1-5 was the most similar, of which the abundance of Proteobacteria gradually increased with the prolonged feeding time of artificial diet, and the abundance of Firmicutes showed an opposite trend. At the genus level, Pseudomonas, Enterococcus, Sphingomonadaceae unclassified and Ralstonia were the dominant bacteria in all groups. The abundance of the intestinal bacterial genus Enterococcus showed the largest difference in B. mori under different rearing patterns, being the highest in Art1-2 (40.9%) and decreasing gradually with the prolonged feeding time of artificial diet, and only accounting for 0.02% in Art1-5. Most of the key intestinal bacteria in B. mori were related to cocoon quality. Coxiella showed a significantly positive correlation with pupal weight, and a significantly negative correlation with cocoon layer weight. Staphylococcus was significantly negatively correlated with the pupal weight, and Enterococcus was significantly positively correlated with cocoon layer weight. 【Conclusion】The rearing pattern of Art1-2 is the closest to that of Mul1-5, which can provide reference for rearing B. mori with artificial diet partially replacing mulberry leaves. The structure of intestinal bacteria significantly change when mulberry leaves are partially replaced by artificial diet, and the significant decrease in the abundance of Enterococcus and increase of Pseudomonas may be related to the weak physique of B. mori fed with artificial diet for all instars. Some of the key bacteria are significantly correlated with pupal weight and cocoon layer weight of B. mori, although the mechanism needs further study.

Key words: Bombyx mori, artificial diet, mulberry leaves, intestinal bacteria, cocoon production performance