杜仲梦尼夜蛾幼虫肠道可培养细菌的组成及其功能

doi:10.11707/j.1001-7488.20201113

Abstract

Abstract:

Objective: Gut microbiota can help host insects to degrade indigestible ingredients and harmful substance in food. The ability to digest and utilize the leaves of Eucomnia ulmoides in Orthosia songi (Lepidoptera: Noctuidae) larvae may be related with gut microbiota. Therefore,this study aims to investigate the gut bacterial composition of larvae in O. songi,so as to reveal the effects of intestinal bacteria on the growth and development,food digestion,nutrition utilization of host insects. Method: The traditional methods of microbial isolation and culture and the sequencing of 16S rRNA gene were used to isolate and identify the gut bacteria from the larvae. The dinitrosalicylic acid (DNS) and HPLC methods were used to verify the degradation of biomacromolecules (cellulose,pectin and starch) and chemical substances in leaves of Eucommia ulmoides by gut bacteria. Then the effects of gut bacteria on the growth and nutritional utilization of larvae were investigated by feeding 1 μg·mL^-1 chloramphenicol,an antibiotic. Result: Ten different bacterial strains were isolated from the larval guts by traditional microbial isolation and culture methods,and their DNA sequences showed that they belonged to ten genera in Proteobacteria,Firmicutes and Actinobacteria,including Brevundimonas,Enterococcus,Pseudomonas,Sphingomonas,Kluyvera,Acinetobacter,Moraxinella,Agrobacterium,Microbacterium, and Arthrobacter. These strains were able to cause degradation to biomacromolecules (cellulose,pectin and starch) in varying degrees,and affect the changes of some substances in Eucommia ulmoides leaves. The feeding test with 1μg/mL chloramphenicol affected the growth and development of larvae,and significantly reduced the pupation rate of larvae and the feeding and digestive ability of the younger larvae. Conclusion: Ten bacterial strains were isolated from the larval guts of O. songi,and they play important roles in the growth and development,food digestion and nutrient utilization of O. songi larvae.

Key words: Orthosia songi, gut bacteria of larvae, isolation of culturable microbe, Lepidoptera

CLC Number:

S718.83

Tongtong Guo,Mingqiang Zhu,Jinpeng Li,Xinrong Guo,Hong He. Composition and Functions of Cultural Bacteria in the Larval Guts of Orthosia songi(Lepidoptera: Noctuidae)[J]. Scientia Silvae Sinicae, 2020, 56(11): 124-133.

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菌株编号 Strain code	Blast结果中一致性最高的菌株(GenBank登录号) (一致性) BLAST with the highest identity (GenBank accession No.) (Identity)	序列登录号 Accession No.
LT1	肠球菌属Enterococcus sp. (KJ124182.1)(99%)	MK571186
NT2	土壤杆菌属Agrobacterium sp. (KT230833.1)(99%)	MK571187
NA3	不动杆菌属Acinetobacter sp. (KY910080.1)(99%)	MK571188
NT4	莫拉菌属Moraxella sp.(FJ380954.1)(99%)	MK571189
NA5	假单胞菌属Pseudomonas sp. (JX675235.1)(99%)	MK571190
NA6	克吕沃尔菌属Kluyvera sp. (GQ915083.1)(99%)	MK571191
NT7	短波单胞菌属Brevundimonas sp. (KF735808.1)(99%)	MK571192
NA8	鞘氨醇单胞菌属Sphingomonas sp. (EU130999.1)(99%)	MK571193
NA9	节杆菌属Arthrobacter sp.(KU060821.1)(99%)	MK571194
NT10	微杆菌属Microbacterium sp. (EU558969.1)(99%)	MK571195

幼虫龄期 Larval instars	发育历期 Developmental duration/d		头宽 Head width/mm		体长 Body length/mm		体质量 Body mass/mg
幼虫龄期 Larval instars	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK
1龄 1st instar	4.10±0.84a	3.93±0.87a	0.4±0.11a	0.42±0.12a	2.94±0.44a	3.52±0.37a	0.48±0.13a	0.58±0.18a
2龄 2nd instar	2.87±0.94a	2.73±0.74a	0.57±0.12a	0.66±0.29a	4.88±1.08a	5.74±1.05a	1.28±0.40a	2.24±0.44a
3龄 3rd instar	2.70±0.70a	2.57±0.73a	0.56±0.06a	0.87±0.06a	6.87±0.97a	7.48±0.66a	8.10±4.41a	9.07±3.03a
4龄 4th instar	3.80±0.76a	3.57±0.78a	1.03±0.08a	1.13±0.09a	11.00±1.35a	11.05±1.18a	24.85±3.55a	25.94±5.38a
5龄 5th instar	3.80±0.85a	3.47±0.71a	1.70±0.14a	1.74±0.16a	14.93±2.28a	15.09±1.83a	78.79±35.36a	83.07±31.95a
6龄 6th instar	5.60±0.86a	5.43±1.22a	2.47±0.05a	2.48±0.05a	21.92±2.46a	22.24±1.76a	180.02±44.06a	190.9±35.16a

幼虫龄期 Larval instars	幼虫相对生长率RGR		食物利用率ECI		食物转化率ECD		消耗指数CI		近似消耗率AD
幼虫龄期 Larval instars	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK	抗生素处理 Antibiotics treatment	CK
1龄 1st instar	0.34±0.05b	0.49±0.07a	0.08±0.05b	0.21±0.02a	0.08±0.02b	0.42±0.01a	4.45±2.29a	4.4±2.23a	0.94±0.25a	0.91±0.27a
2龄 2nd instar	0.17±0.07b	0.57±0.07a	0.07±0.07b	0.31±0.01a	0.12±0.04b	0.58±0.07a	2.44±1.25a	1.86±0.65a	0.58±0.12a	0.53±0.13a
3龄 3rd instar	0.15±0.04b	0.48±0.02a	0.15±0.00b	0.47±0.04a	0.25±0.02b	0.83±0.25a	1.03±0.45a	1.02±0.47a	0.59±0.25a	0.57±0.25a
4龄 4th instar	0.07±0.01b	0.19±0.02a	0.08±0.01b	0.2±0.01a	0.19±0.00b	0.49±0.03a	0.88±0.40a	0.86±0.38a	0.44±0.00a	0.42±0.00a
5龄 5th instar	0.11±0.01a	0.13±0.01a	0.1±0.01a	0.16±0.00a	0.18±0.02b	0.29±0.05a	1.06±0.55a	0.8±0.34a	0.59±0.00a	0.57±0.03a
6龄 6th instar	0.04±0.01a	0.09±0.01a	0.04±0.00a	0.09±0.01a	0.26±0.01a	0.43±0.01a	1.18±0.35a	0.97±0.44a	0.24±0.00a	0.21±0.01a

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Composition and Functions of Cultural Bacteria in the Larval Guts of Orthosia songi(Lepidoptera: Noctuidae)

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