Bt杀虫蛋白处理后二化螟幼虫中肠细菌群落的变化

doi:10.16380/j.kcxb.2016.03.005

昆虫学报 ›› 2016, Vol. 59 ›› Issue (3): 292-300.doi: 10.16380/j.kcxb.2016.03.005

Bt杀虫蛋白处理后二化螟幼虫中肠细菌群落的变化

李振^1,2, 韩兰芝¹, 刘玉娣^1,*, 侯茂林¹

（1. 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193; 2. 福建农林大学植物保护学院, 福州 350002）

出版日期:2016-03-20 发布日期:2016-03-20
作者简介:李振, 女, 1992年1月生, 河南太康人, 硕士研究生, 研究方向为昆虫生化与分子生物学, E-mail: lz19920110@126.com

Change in the diversity of bacterial community in larval midguts of the striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae), after treatment with Bacillus thuringiensis insecticidal proteins

LI Zhen^1,2, HAN Lan-Zhi¹, LIU Yu-Di^1,*, HOU Mao-Lin¹

(1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2. College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, China)

Online:2016-03-20 Published:2016-03-20

摘要/Abstract

摘要： 【目的】为探讨经Bt杀虫蛋白处理后二化螟 Chilo suppressalis (Walker)幼虫中肠细菌群落的差异。【方法】本研究对采自北京（BJ）和福州（FZ）2个地区在室内分别经过未用Bt杀虫蛋白和使用Bt杀虫蛋白（Cry1Ab, Cry1Ac和Cry1Ca）多代汰选条件下的二化螟幼虫中肠进行解剖，采用变性梯度凝胶电泳（DGGE）和Illumina MiSeq 技术测序平台对3个处理组（ BJCry1Ab, BJCry1Ac和FZ1Ca）和2个对照组(BJCK和FZCK)中肠细菌的16S rDNA V3可变区进行电泳检测和高通量测序。【结果】DGGE图谱显示，5个不同处理组的细菌不仅丰富度存在差异，同时同种细菌在不同处理组中的比例也存在差异。高通量测序结果表明，优势菌为厚壁菌门（Firmicutes）的肠球菌属 Enterococcus，其次为乳杆菌属 Lactobacillus 和芽孢杆菌属 Bacillus , 以及变形菌门（Proteobacteria）、绿弯菌门（Chloroflexi）和拟杆菌门（Bacteroidetes）。同一地区的二化螟种群，Bt杀虫蛋白处理组（BJCry1Ab, BJCry1Ac和FZ1Ca）的二化螟幼虫与对照组（BJCK和FZCK）相比，优势菌肠球菌属 Enterococcus 比重均有所增加，而乳杆菌属 Lactobacilluss 所占比重均有所降低。北京和福州这2个地区未用Bt杀虫蛋白处理的对照组之间肠道菌落的结构也存在一定差异。【结论】经Bt杀虫蛋白处理后二化螟幼虫中肠细菌群落的丰富度出现变化，推测可能与二化螟取食不同Bt杀虫蛋白、地理位置差异以及饲养代数不同有关。

关键词: 二化螟, 中肠, 变性梯度凝胶电泳, 16S rDNA文库, 细菌群落多样性, 苏云金芽孢杆菌

Abstract: 【Aim】 To clarify the change in the bacterial community in larval midguts of the striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae) after treatment with Bacillus thuringiensis insecticidal proteins. 【Methods】 Midguts were dissected from C. suppressalis larvae that were collected from Beijing (BJ) and Fuzhou (FZ) of China and successively reared with artificial diets without Bt insecticidal proteins and with Bt insecticidal proteins (Cry1Ab, Cry1Ac and Cry1Ca), respectively. Degeneration gradient gel electrophoresis (DGGE) and Illumina MiSeq platform were employed to detect the bacterial community in these midguts and to compare its difference among different treatments (BJCK, BJCry1Ab,BJCry1Ac, FZCK and FZ1Ca) of the two populations. 【Results】 The DGGE profiles demonstrated that there was difference in bacterial abundance in larval midguts of C. suppressalis among different treatments. Analyses on DNA sequence of 16S rDNA libraries indicated that among the midgut bacteria Enterococcus bacteria of Firmicutes was the most dominant, followed by bacteria of Lactobacillus and Bacillus (Firmicutes), Proteobacteria, Chloroflexi, and Bacteroidetes. The proportions of the dominant Enterococcus bacteria in the larvae treated with Bt insecticidal proteins (BJCry1Ab, BJCry1Ac and FZ1Ca) were higher than those in the larvae without treatment with Bt (BJCK and FZCK), but the proportions of Lactobacillus bacteriawere lower in the larvae treated with Bt insecticidal proteins. The bacterial abundance also differed between BJCK and FZCK. 【Conclusion】 The results suggest that the bacterial abundance in larval midguts of C. suppressalis changes after treatment with Bt insecticidal proteins, which may be related to the difference in Bt insecticidal proteins, geographical sites and successive rearing time.

Key words: Chilo suppressalis, midgut, PCR-DGGE, 16S rDNA libraries, bacterial, Bacillus thuringiensis

李振, 韩兰芝, 刘玉娣, 侯茂林. Bt杀虫蛋白处理后二化螟幼虫中肠细菌群落的变化[J]. 昆虫学报, 2016, 59(3): 292-300.

LI Zhen, HAN Lan-Zhi, LIU Yu-Di, HOU Mao-Lin. Change in the diversity of bacterial community in larval midguts of the striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae), after treatment with Bacillus thuringiensis insecticidal proteins[J]. Acta Entomologica Sinica, 2016, 59(3): 292-300.

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http://www.insect.org.cn/CN/Y2016/V59/I3/292

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Bt杀虫蛋白处理后二化螟幼虫中肠细菌群落的变化

Change in the diversity of bacterial community in larval midguts of the striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae), after treatment with Bacillus thuringiensis insecticidal proteins

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