Diversity of Midgut Microbial Community of ChilosuppressalisExposed to Chlorobenzamide

doi:10.16819/j.1001-7216.2020.0101

Abstract

Abstract:

【Objective】We aim toilluminatethestructural changes in the midgut microbial community of Chilosuppressalis exposed to chlorobenzamide.【Method】Macrogenomic sequencing and in vitro culturewere used to analyze the diversity of midgut microbiotaof Chilosuppressalistreatedwith different concentrations (100, 200, 400µg/mL) of chlorobenzamide.【Result】The results showed that chlorobenzamide treatmentdecreased the abundance anddiversityof the midgut bacteria of Chilosuppressalis,and the OTU number and specific OTU number of the intestinal bacteria of treated population were lower than those of the control;the proportion of Morganella, Provincia and Proteus in the chlorobenzamide treated population increased significantly. Meanwhile, the result of in vitro culture indicated besides enterobacter,Lelliottinimipressuralis, Leclerciadecarboxylata, Pantoeaagglomeranswere isolated from chlorobenzamide treated Chilosuppressalis population. COG classification and KEGG analysis didn’t show significant difference in number and relative abundance of functional genes among control and three chlorobenzamide treated populations.【Conclusion】The study had clarified the diversity of intestinal bacteria and the relationshipbetween gutbacteria of Chilosuppressalis andthe formation of chlorobenzamide resistance, and will lay a scientific basis for the role of intestinal bacteria in resistance of Chilosuppressalistochlorobenzamide.

Key words: Chilosuppressalis, chlorobenzamide, macrogenomic sequencing, midgut bacteria, microbial diversity

摘要： 目的明确氯虫苯甲酰胺处理下二化螟中肠细菌类微生物多样性的变化。方法综合运用宏基因组测序以及传统微生物分离纯化的方法分析不同浓度氯虫苯甲酰胺处理的二化螟种群中肠微生物的多样性。结果不同浓度(100、200、400µg/mL)氯虫苯甲酰胺处理后二化螟中肠细菌类微生物丰度及多样性降低,处理种群中肠细菌类微生物的OTU数值、特有的OTU数值均低于对照;摩根氏菌属(Morganella)、普罗威登斯菌属(Providencia)以及变形菌属(Proteus)在处理种群中的比例明显升高;同时培养结果显示氯虫苯甲酰胺处理种群获得的菌株除肠杆菌属(Enterobacter)外,还存在超压莱略特氏菌(Lelliottianimipressuralis)、非脱羧勒菌(Leclerciaadecarboxylata)、成团泛菌(Pantoeaagglomerans)等其他菌属。COG数据库对比与KEGG分析显示,对照与处理二化螟种群的各功能分类基因数量及相对丰度无明显差异。结论解析了氯虫苯甲酰胺胁迫下二化螟中肠细菌类微生物的多样性。这些结果为进一步研究中肠细菌在二化螟对氯虫苯甲酰胺的抗药性奠定基础。

关键词: 二化螟, 氯虫苯甲酰胺：宏基因组, 中肠细菌, 微生物多样性

CLC Number:

Juefeng ZHANG, Qin ZHANG, Fang LI, Haiying ZHONG, Jianming CHEN. Diversity of Midgut Microbial Community of ChilosuppressalisExposed to Chlorobenzamide[J]. Chinese Journal OF Rice Science, 2020, 34(6): 586-594.

张珏锋, 张琴, 李芳, 钟海英, 陈建明. 氯虫苯甲酰胺胁迫下二化螟中肠细菌类微生物的多样性[J]. 中国水稻科学, 2020, 34(6): 586-594.

Figures/Tables 6

References 31

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样本 Sample	OUT (97%)	Ace指数 Ace index	Chao指数 Chao index	测序深度指数 Sequencing depth	香农指数 Shannon index	辛普森指数 Simpson index
100_24h	333	442(407,494)	405(377,452)	0.99	2.19(2.18, 2.21)	0.20(0.20, 0.20)
100_48h	432	758(693,838)	640(571,742)	0.99	2.39(2.37, 2.40)	0.17(0.17, 0.17)
100_72h	580	718(680,771)	745(690,827)	0.99	2.98(2.96, 2.99)	0.13(0.13, 0.13)
200_24h	412	512(481,557)	500(467,553)	0.99	2.02(2.00, 2.04)	0.32(0.32, 0.32)
200_48h	357	415(394,448)	407(385,447)	0.99	2.49(2.47, 2.50)	0.16(0.16, 0.16)
200_72h	639	742(713,782)	730(698,778)	0.99	3.02(3.01, 3.04)	0.13(0.12, 0.13)
400_24h	474	578(547,623)	569(535,623)	0.99	2.70(2.68, 2.71)	0.12(0.12, 0.12)
400_48h	751	901(863,952)	893(849,956)	0.99	3.31(3.29, 3.33)	0.09(0.09, 0.09)
400_72h	321	539(473,634)	557(473,690)	0.99	2.22(2.20, 2.23)	0.16(0.15, 0.16)
CK_24h	1003	1127(1096,1169)	1123(1085,1177)	0.99	3.91(3.89, 3.93)	0.07(0.07, 0.07)
CK_48h	718	782(763,811)	780(757,817)	0.99	3.38(3.36, 3.40)	0.08(0.09, 0.09)
CK_72h	650	791(754,842)	769(730,827)	0.99	3.38(3.36, 3.39)	0.08(0.08, 0.08)

样本 Sample	OUT (97%)	Ace指数 Ace index	Chao指数 Chao index	测序深度指数 Sequencing depth	香农指数 Shannon index	辛普森指数 Simpson index
100_24h	333	442(407,494)	405(377,452)	0.99	2.19(2.18, 2.21)	0.20(0.20, 0.20)
100_48h	432	758(693,838)	640(571,742)	0.99	2.39(2.37, 2.40)	0.17(0.17, 0.17)
100_72h	580	718(680,771)	745(690,827)	0.99	2.98(2.96, 2.99)	0.13(0.13, 0.13)
200_24h	412	512(481,557)	500(467,553)	0.99	2.02(2.00, 2.04)	0.32(0.32, 0.32)
200_48h	357	415(394,448)	407(385,447)	0.99	2.49(2.47, 2.50)	0.16(0.16, 0.16)
200_72h	639	742(713,782)	730(698,778)	0.99	3.02(3.01, 3.04)	0.13(0.12, 0.13)
400_24h	474	578(547,623)	569(535,623)	0.99	2.70(2.68, 2.71)	0.12(0.12, 0.12)
400_48h	751	901(863,952)	893(849,956)	0.99	3.31(3.29, 3.33)	0.09(0.09, 0.09)
400_72h	321	539(473,634)	557(473,690)	0.99	2.22(2.20, 2.23)	0.16(0.15, 0.16)
CK_24h	1003	1127(1096,1169)	1123(1085,1177)	0.99	3.91(3.89, 3.93)	0.07(0.07, 0.07)
CK_48h	718	782(763,811)	780(757,817)	0.99	3.38(3.36, 3.40)	0.08(0.09, 0.09)
CK_72h	650	791(754,842)	769(730,827)	0.99	3.38(3.36, 3.39)	0.08(0.08, 0.08)

分类 Classification	对照CK		100µg/mL		200µg/mL		400µg/mL
分类 Classification	菌株名称 Strain	数量 Quantity	菌株名称 Strain	数量 Quantity	菌株名称 Strain	数量 Quantity	菌株名称 Strain	数量 Quantity
肠杆菌属 Enterobacter	霍氏肠杆菌 Enterobacter hormaechei	4					霍氏肠杆菌 Enterobacter hormaechei	3
	阿氏肠杆菌YT Enterobacter asburiae strain YT	1					阿氏肠杆菌YT Enterobacter asburiaestrain YT	2
	肠杆菌属 CZBSY4Enterobacter sp. CZBSY4	4					肠杆菌属 CZBSY4 Enterobacter sp. CZBSY4	1
	肠杆菌属 09-M1 Enterobacter sp. 09-M1	2
	肠杆菌属 XM Enterobacter sp. strain XM	3
	阴沟肠杆菌 Enterobacter cloacaether	1
	路氏肠杆菌 Enterobacter ludwigii	2
			肠杆菌属 S4(2015) Enterobacter sp. S4(2015)	4			肠杆菌属 S4(2015) Enterobacter sp. S4(2015)	1
			产气肠杆菌	2	产气肠杆菌	1	产气肠杆菌	1
			Enterobacter roggenkampii Enterobacter aerogenes	1	Enterobacter roggenkampii Enterobacter aerogenes	3	Enterobacter roggenkampii Enterobacter aerogenes	1
			森肠杆菌 Enterobacter mori	1	森肠杆菌 Enterobacter mori	2	森肠杆菌 Enterobacter mori	1
			河生肠杆菌 Enterobacter amnigenus	3	河生肠杆菌 Enterobacter amnigenus	4
			神户肠杆菌 Enterobacter kobei	1	神户肠杆菌 Enterobacter kobei	1
			阿氏肠杆菌 Enterobacter asburiae	3
其他 Others	非脱羧埃希氏菌 Leclerciaadecarboxylata	2	非脱羧埃希氏菌 Leclerciaadecarboxylata	1	非脱羧埃希氏菌 Leclerciaadecarboxylata	2	非脱羧埃希氏菌 Leclerciaadecarboxylata	5
			成团泛菌 Pantoeaagglomerans	1	成团泛菌 Pantoeaagglomerans	3	成团泛菌 Pantoeaagglomerans	2
			泛菌属DAP16 Pantoea sp. DAP16	1	泛菌属DAP16 Pantoea sp. DAP16	2	泛菌属DAP16 Pantoea sp. DAP16	2
			克雷伯氏菌属 Klebsiella sp.	1	克雷伯氏菌属 Klebsiella sp.	1
			超压莱略特氏菌 Lelliottianimipressuralis	1	超压莱略特氏菌 Lelliottianimipressuralis	1
			ɤ-变形菌门细菌 Gammaproteobacteria bacterium strain	3	ɤ-变形菌门细菌 Gammaproteobacteria bacterium strain	1
			不可培养细菌 Uncultured bacterium	3	不可培养细菌 Uncultured bacterium	3	不可培养细菌 Uncultured bacterium	1

分类 Classification	对照CK		100µg/mL		200µg/mL		400µg/mL
分类 Classification	菌株名称 Strain	数量 Quantity	菌株名称 Strain	数量 Quantity	菌株名称 Strain	数量 Quantity	菌株名称 Strain	数量 Quantity
肠杆菌属 Enterobacter	霍氏肠杆菌 Enterobacter hormaechei	4					霍氏肠杆菌 Enterobacter hormaechei	3
	阿氏肠杆菌YT Enterobacter asburiae strain YT	1					阿氏肠杆菌YT Enterobacter asburiaestrain YT	2
	肠杆菌属 CZBSY4Enterobacter sp. CZBSY4	4					肠杆菌属 CZBSY4 Enterobacter sp. CZBSY4	1
	肠杆菌属 09-M1 Enterobacter sp. 09-M1	2
	肠杆菌属 XM Enterobacter sp. strain XM	3
	阴沟肠杆菌 Enterobacter cloacaether	1
	路氏肠杆菌 Enterobacter ludwigii	2
			肠杆菌属 S4(2015) Enterobacter sp. S4(2015)	4			肠杆菌属 S4(2015) Enterobacter sp. S4(2015)	1
			产气肠杆菌	2	产气肠杆菌	1	产气肠杆菌	1
			Enterobacter roggenkampii Enterobacter aerogenes	1	Enterobacter roggenkampii Enterobacter aerogenes	3	Enterobacter roggenkampii Enterobacter aerogenes	1
			森肠杆菌 Enterobacter mori	1	森肠杆菌 Enterobacter mori	2	森肠杆菌 Enterobacter mori	1
			河生肠杆菌 Enterobacter amnigenus	3	河生肠杆菌 Enterobacter amnigenus	4
			神户肠杆菌 Enterobacter kobei	1	神户肠杆菌 Enterobacter kobei	1
			阿氏肠杆菌 Enterobacter asburiae	3
其他 Others	非脱羧埃希氏菌 Leclerciaadecarboxylata	2	非脱羧埃希氏菌 Leclerciaadecarboxylata	1	非脱羧埃希氏菌 Leclerciaadecarboxylata	2	非脱羧埃希氏菌 Leclerciaadecarboxylata	5
			成团泛菌 Pantoeaagglomerans	1	成团泛菌 Pantoeaagglomerans	3	成团泛菌 Pantoeaagglomerans	2
			泛菌属DAP16 Pantoea sp. DAP16	1	泛菌属DAP16 Pantoea sp. DAP16	2	泛菌属DAP16 Pantoea sp. DAP16	2
			克雷伯氏菌属 Klebsiella sp.	1	克雷伯氏菌属 Klebsiella sp.	1
			超压莱略特氏菌 Lelliottianimipressuralis	1	超压莱略特氏菌 Lelliottianimipressuralis	1
			ɤ-变形菌门细菌 Gammaproteobacteria bacterium strain	3	ɤ-变形菌门细菌 Gammaproteobacteria bacterium strain	1
			不可培养细菌 Uncultured bacterium	3	不可培养细菌 Uncultured bacterium	3	不可培养细菌 Uncultured bacterium	1