Objective To screen for bacteria against Beauveria bassiana from the intestinal bacteria of Musca domestica larvae. Methods The traditional isolation and culture methods were used to isolate intestinal bacteria from M. domestica larvae, and the bacteria were identified by bacterial colony morphology and 16S rDNA sequencing. The spore germination test was used to screen for the intestinal bacteria of M. domestica inhibiting the spore germination of B. bassiana. The plate confrontation test was used to screen for the intestinal bacteria of M. domestica inhibiting the mycelial growth of B. bassiana. Results Thirteen species of bacteria were isolated from the intestines of M. domestica larvae by the traditional isolation and culture methods. The spore germination test showed that only Pseudomonas aeruginosa significantly inhibited the spore germination of B. bassiana, with a germination rate of 17%, while the spore germination rates for other bacteria and control group were above 90%. The plate confrontation test showed that P. aeruginosa significantly inhibited the mycelial growth of B. bassiana, and that the mean diameter of the inhibitory zone on the plate was about 30 mm, while other intestinal bacteria from M. domestica larvae did not inhibit the mycelial growth of B. bassiana. Conclusion Pseudomonas aeruginosa in the intestines of M. domestica larvae can inhibit the spore germination and mycelial growth of B. bassiana, and this may be one reason why M. domestica larvae are immune to B. bassiana.
[1] Bextine BR,Thorvilson HG. Field applications of bait-formulated Beauveria bassiana alginate pellets for biological control of the red imported fire ant (Hymenoptera:Formicidae)[J]. Environ Entomol,2002,31(4):746-752. DOI:10.1603/0046-225X-31.4.746.
[2] 张璐璐,吴圣勇,王帅宇,等. 防治蓟马的球孢白僵菌SDDZ-9菌株液体发酵工艺优化[J]. 中国农业科学,2015,48(15):2985-2994. DOI:10.3864/j.issn.0578-1752.2015.15.008. Zhang LL,Wu SY,Wang SY,et al. Optimization of fermentation process of Beauveria bassiana of SDDZ-9 against Frankliniella occidenta[J]. Sci Agric Sin,2015,48(15):2985-2994. DOI:10.3864/j.issn.0578-1752.2015.15.008.
[3] Ishii M,Kanuka H,Badolo A,et al. Proboscis infection route of Beauveria bassiana triggers early death of Anopheles mosquito[J]. Sci Rep,2017,7(1):3476. DOI:10.1038/s41598-017-03720-x.
[4] 庄桂芬,黄振东,薛志静,等. 球孢白僵菌CF08株在家蝇成虫间的水平传播实验[J]. 中国媒介生物学及控制杂志,2017,28(2):131-134. DOI:10.11853/j.issn.1003.8280.2017.02.008. Zhuang GF,Huang ZD,Xue ZJ,et al. Horizontal transmission of Beauveria bassiana CF08 strain (Hypocreales:Cordycipitaceae) in Musca domestica (Diptera:Muscidae) adults[J]. Chin J Vector Biol Control,2017,28(2):131-134. DOI:10.11853/j.issn.1003.8280.2017.02.008.
[5] 谭清,庞仁乙,高熹,等. 蜡蚧轮枝菌分生孢子在不同虫态棕尾别麻蝇体表的附着能力及与寄主体表结构的关系[J]. 昆虫学报,2014,57(10):1245-1252. Tan Q,Pang RY,Gao X,et al. Adhesion capacity of Lecanicillium lecanii conidia on body surface of Boettcherisca peregrina (Diptera:Sarcophagidae) at different developmental stages and its relationship with surface structures of host body[J]. Acta Entomol Sin,2014,57(10):1245-1252.
[6] 谭清,高熹,庞仁乙,等. 家蝇不同虫态附着蜡蚧轮枝菌分生孢子能力及与其体表结构的关系[J]. 南方农业学报,2015,46(2):241-249. DOI:10.3969/jissn.2095-1191.2015.2.241. Tan Q,Gao X,Pang RY,et al. Adhesion capacities of Lecanicillium lecanii conidia on different insect states of Musca domestica and the relationship with their surface structures[J]. J Southern Agric,2015,46(2):241-249. DOI:10.3969/jissn.2095-1191.2015.2.241.
[7] Lu DD,Geng T,Hou CX,et al. Bombyx mori cecropin A has a high antifungal activity to entomopathogenic fungus Beauveria bassiana[J]. Gene,2016,583(1):29-35. DOI:10.1016/j.gene.2016.02.045.
[8] 张慧,吴圣勇,王帅宇,等. 葱蝇成虫防御类酶和血细胞对球孢白僵菌的防御作用[J]. 中国生物防治学报,2017,33(2):198-205. DOI:10.16409/j.cnki.2095-039x.2017.02.009. Zhang H,Wu SY,Wang SY,et al. Effect of Beauveria bassiana on the activity of defense enzymes and cellular defense response of adult of Delia antiqua (Meigen)[J]. Chin J Biol Control,2017,33(2):198-205. DOI:10.16409/j.cnki.2095-039x.2017. 02.009.
[9] 汤历,廉婕,陆小军,等. 德国小蠊肠道细菌抗真菌的初步研究[J]. 昆虫天敌,2005,27(3):140-144. DOI:10.3969/j.issn. 1674-0858.2005.03.007. Tang L,Lian J,Lu XJ,et al. Antifungal activity of intestinal bacteria of Blattella germanica[J]. Nat Enemies Insects,2005,27(3):140-144. DOI:10.3969/j.issn.1674-0858.2005.03.007.
[10] 黄艳红,张凡,付荣恕,等. 德国小蠊肠道菌群分离及抗真菌活性分析[J]. 中国公共卫生,2011,27(1):78-79. DOI:10.11847/zgggws2011-27-01-37. Huang YH,Zhang F,Fu RS,et al. Isolation of gut Microbiotas of Blattella germanica and analysis of antifungal activity[J]. Chin J Public Health,2011,27(1):78-79. DOI:10.11847/zgggws2011-27-01-37.
[11] Zhang F,Huang YH,Liu SZ,et al. Pseudomonas reactans,a bacterial strain isolated from the intestinal flora of Blattella germanica with anti-Beauveria bassiana activity[J]. Environ Entomol,2013,42(3):453-459. DOI:10.1603/EN12347.
[12] 黄振东,万晴,庄桂芬,等. 球孢白僵菌CF08株对丽蝇蛹集金小蜂的安全性评价[J]. 中国媒介生物学及控制杂志,2018,29(3):250-253. DOI:10.11853/j.issn.1003.8280.2018.03.008. Huang ZD,Wan Q,Zhuang GF,et al. Safety evaluation of Beauveria bassiana CF08 strain on Nasonia vitripennis[J]. Chin J Vector Biol Control,2018,29(3):250-253. DOI:10.11853/j.issn.1003.8280.2018.03.008.
[13] Wang CS,Duan ZB,St. Leger RJ. MOS1 osmosensor of Metarhizium anisopliae is required for adaptation to insect host hemolymph[J]. Eukaryot Cell,2008,7(2):302-309. DOI:10.1128/EC.00310-07.
[14] 魏蕾. 采用孢子萌发法检测蔬菜灰霉病菌的抗药性[J]. 北京农业科学,2000,18(5):19-21. Wei L. The resistance of Botrytis cinerea of vegetables was used for detections by spore germination[J]. Beijing Agric Sci,2000,18(5):19-21.
[15] 杨小军,倪汉文,杨立军,等. 采用孢子萌发法测定化合物对黄瓜白粉病菌的生物活性[J]. 植物保护,2007,33(1):75-77. DOI:10.3969/j.issn.0529-1542.2007.01.022. Yang XJ,Ni HW,Yang LJ,et al. A novel bioassay method-determining the bioactivity of chemical compounds to cucumber powdery mildew by spore germination[J]. Plant Prot,2007,33(1):75-77. DOI:10.3969/j.issn.0529-1542.2007.01.022.
[16] Kostylev M,Kim DY,Smalley NE,et al. Evolution of the Pseudomonas aeruginosa quorum-sensing hierarchy[J]. Proc Natl Acad Sci USA,2019,116(14):7027-7032. DOI:10.1073/pnas.1819796116.
[17] 李苓苓. 铜绿假单胞菌中影响吩嗪合成基因操纵子phzA1的调节基因的研究[D]. 西安:西北大学,2005. Li LL. The expression and regulation of phzAl operon in P. aeruginosa[D]. Xi'an:Northwestern University,2005.
[18] 陈明明. 家蝇抗真菌肽MAF-1A的分子改良及抗白色念珠菌活性的研究[D]. 贵阳:贵州医科大学,2016. Chen MM. Study on the molecular modification and anti-Canidia albicans activity of Musca domestica antifungal peptide-1A (MAF-1A)[D]. Guiyang:Guizhou Medical University,2016.
