| 草地贪夜蛾肠道细菌分离鉴定与功能初探 |
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| 引用本文： | 陈亚平,杜鄂巍,李亚红,鲁智慧,李浩,桂富荣.草地贪夜蛾肠道细菌分离鉴定与功能初探[J].南方农业学报,2022,53(3):821-829. |
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| 作者姓名： | 陈亚平 杜鄂巍 李亚红 鲁智慧 李浩 桂富荣 |
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| 作者单位： | 1. 云南农业大学植物保护学院/省部共建云南生物资源保护与利用国家重点实验室, 云南昆明 650201;2云南省植保植检站, 云南昆明 650034 |
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| 摘 要： | 【目的】明确草地贪夜蛾幼虫肠道细菌的种类和功能，以诠释肠道细菌对草地贪夜蛾寄主植物适应性的影响，为揭示草地贪夜蛾的寄主适应机制及进一步预测其寄主谱扩张趋势提供依据。【方法】采用传统的微生物分离纯化方法对草地贪夜蛾肠道细菌进行分离纯化，对分离获得的细菌菌株进行16S rRNA序列同源性分析鉴定；利用筛选培养基对产生纤维素酶、木聚糖酶、果胶酶及对苯酚有代谢能力的菌株进行初筛，并进一步用DNS法测定相关菌株产纤维素酶、木聚糖酶和果胶酶的酶活性，用含苯酚的无机盐培养基培养菌株，检测菌株的苯酚降解效率。【结果】共分离获得45株细菌菌株，经同源序列比对分析，45株细菌菌株分属于3门5属8种，分别是厚壁菌门（Firmicutes）的葡萄球菌属（Staphylococcus）和芽孢杆菌属（Bacillus），变形菌门（Proteobacteria）的克雷伯氏菌属（Klebsiella）和不动杆菌属（Acinetobacter），放线菌门（Acinobacteria）的短杆菌属（Curtobacterium），其中克雷伯氏菌属的丰度最高。45株菌株中有产纤维素酶菌株11株，酶活力最高的是变栖克雷伯氏菌菌株K3，为0.105±0.007 U/mL；产木聚糖酶菌株10株，酶活力最高的是枯草芽孢杆菌菌株B9，为1.090±0.468 U/mL；产果胶酶菌株5株，酶活力最高的是变栖克雷伯氏菌菌株K27，为0.193±0.047 U/mL；降解苯酚的菌株9株，降解速率最高的是沙福芽孢杆菌菌株B8，为（0.347±0.042）%。【结论】草地贪夜蛾幼虫肠道细菌的产酶菌株多样性较高，推测这是导致草地贪夜蛾寄主谱广，对寄主为害严重的原因之一。
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| 关 键 词： | 草地贪夜蛾 肠道细菌 分离鉴定 酶活测定 苯酚降解 |
| 收稿时间： | 2021-11-02 |
Isolation,identification and functional analysis of intestinal bacteria of Spodoptera frugiperda |
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| CHEN Ya-ping,DU E-wei,LI Ya-hong,LU Zhi-hui,LI Hao,GUI Fu-rong.Isolation,identification and functional analysis of intestinal bacteria of Spodoptera frugiperda[J].Journal of Southern Agriculture,2022,53(3):821-829. |
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| Authors: | CHEN Ya-ping DU E-wei LI Ya-hong LU Zhi-hui LI Hao GUI Fu-rong |
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| Institution: | 1. Plant Protection College, Yunnan Agricultural University/State Key Laboratory for Conservation and Utilization of Bioresources in Yunnan, Kunming, Yunnan 650201, China;2. Yunnan Plant Protection and Plant Inspection Station, Kunming, Yunnan 650034, China |
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| Abstract: | 【Objective】To clarify the species and functions of intestinal bacteria of Spodoptera frugiperda and explain the effects of intestinal bacteria on the adaptability of S. frugiperda to host plants,so as to provide a theoretical basis for revealing the host adaptation mechanism of S. frugiperda and predict the expansion trend of host spectrum.【Method】The intestinal bacteria of S. frugiperda were isolated and purified using the microbial isolation and purification methods,and the 16S rRNA gene sequence homology analysis was performed on the isolated bacterial strains. The strains that produced cellulase,xylanase,pectinase and metabolized phenol were preliminarily screened. And the cellulase,xylanase,pectinase production activities and metabolism of phenol were analyzed through DNS method. The strains were cultured in phenolcontaining inorganic salt medium to detect their phenol degradation efficiency.【Result】A total of 45 bacterial strains were isolated by comparative homologous sequence analysis,and they belonged to 5 genera of Firmicutes,Proteobacteria,and Acinobacteria,namely,Staphylococcus,Bacillus,Klebsiella,Acinetobacter and Colletotrichum,among which Klebsiella had the highest abundance. Among the 45 strains,11 strains could produce cellulase and K3 of Klebsiella had the highest enzyme activity of 0.105±0.007 U/mL;10 strains could produce xylanase and B9 of Bacillus had the highest enzyme activity of 1.090±0.468 U/mL;5 strains could produce pectinase K27 of Klebsiella had the highest enzyme activity of 0.193±0.047 U/mL;9 strains could degrade phenol and B8 of Bacillus had the highest degradation rate of(0.347±0.042)%.【Conclusion】 Diversity of enzyme-producing strains of intestinal bacteria in S. frugiperda larvae is speculated to be one of the reasons for the wide host spectrum and serious damage to the host. |
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