Culture and Antibiotic Resistance Genes of Endophytic Bacteria in <i>Azolla</i>

CHEN Jian; ZHENG Yiping; CHEN Bin; ZHENG Siping; ZHENG Weiwen

doi:10.19303/j.issn.1008-0384.2021.06.013

Volume 36 Issue 6

Jun. 2021

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(6): 706-712

CHEN J, ZHENG Y P, CHEN B, et al. Culture and Antibiotic Resistance Genes of Endophytic Bacteria in Azolla [J]. Fujian Journal of Agricultural Sciences，2021，36（6）：706−712 doi: 10.19303/j.issn.1008-0384.2021.06.013

Citation:

CHEN J, ZHENG Y P, CHEN B, et al. Culture and Antibiotic Resistance Genes of Endophytic Bacteria in Azolla [J]. Fujian Journal of Agricultural Sciences，2021，36（6）：706−712 doi: 10.19303/j.issn.1008-0384.2021.06.013

Citation:

PDF( 606 KB)

Culture and Antibiotic Resistance Genes of Endophytic Bacteria in Azolla

doi: 10.19303/j.issn.1008-0384.2021.06.013

Biotechnology Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350003, China

Received Date: 2021-03-09
Rev Recd Date: 2021-04-27

Available Online: 2021-06-06

Publish Date: 2021-06-28

Abstract

Abstract

Objective Culture methods for and detection of the antibiotic resistance genes (ARGs) in the endophytic bacteria of Azolla were established to monitor and find means to curtail the drug pollution in aquatic environment. Method Three selected media were used to culture the endophytic bacteria in Azolla microphylla for metagenome sequencing and bioinformatical analysis on ARGs. Result A total of 48.66G clean data was obtained to yield 54 180 predicted ORFs. Through the taxonomic information database NR, 1 712 microbial species were annotated belonging to 38 phyla. On species abundance of the bacteria from the 3 cultures, Proteobacteria accounted for more than 99.14% of all at the phyla level, while at genus level, Herbaspirillum approximately 91% and Burkholderia 1.2%. Blasted with the ARDB database, 3 samples were annotated with 10 genes associated with the resistance to bacitracin and chloramphenicol. Compared to CARD, there were 212 antibiotic related genes (AROs) found in the samples. The genes resistant to antibiotics (AR) accounted for more than 88% of total AROs, while those sensitive to antibiotics (AS) approximately 7% and those resistant to target antibiotics (AT) 4-6%. Conclusion The community structure and ARG types of the endophytic bacteria in Azolla cultured in the 3 different media were basically same.
- Azolla,
- endophytic bacteria,
- antibiotic resistance genes,
- metagenome

FullText(HTML)

References(29)

References

[1]	MARTÍNEZ J L. Antibiotics and antibiotic resistance genes in natural environments [J]. Science, 2008, 321(5887): 365−367. doi: 10.1126/science.1159483
[2]	RAZAVI M, MARATHE N P, GILLINGS M R, et al. Discovery of the fourth mobile sulfonamide resistance gene [J]. Microbiome, 2017, 5(1): 160. doi: 10.1186/s40168-017-0379-y
[3]	高盼盼, 罗义, 周启星, 等. 水产养殖环境中抗生素抗性基因(ARGs)的研究及进展 [J]. 生态毒理学报, 2009, 4(6):770−779. GAO P P, LUO Y, ZHOU Q X, et al. Research advances of antibiotic resistance genes (ARGs) in aquaculture environment [J]. Journal of Ecotoxicology, 2009, 4(6): 770−779.(in Chinese)
[4]	张骞月, 赵婉婉, 吴伟. 水产养殖环境中抗生素抗性基因污染及其研究进展 [J]. 中国农业科技导报, 2015, 17(6):125−134. ZHANG Q Y, ZHAO W W, WU W. Antibiotic resistance gene contamination in aquaculture environment and its research advances [J]. China Agricultural Science and Technology Review, 2015, 17(6): 125−134.(in Chinese)
[5]	胡莹莹, 王菊英, 马德毅. 近岸养殖区抗生素的海洋环境效应研究进展 [J]. 海洋环境科学, 2004, 23(4):76−80. doi: 10.3969/j.issn.1007-6336.2004.04.020 HU Y Y, WANG J Y, MA D Y. Advances on marine environmental effects of antibiotics in inshore aquaculture areas [J]. Marine Environmental Science, 2004, 23(4): 76−80.(in Chinese) doi: 10.3969/j.issn.1007-6336.2004.04.020
[6]	何基兵, 胡安谊, 陈猛, 等. 九龙江河口及厦门污水处理设施抗生素抗性基因污染分析 [J]. 微生物学通报, 2012, 39(5):683−695. HE J B, HU A Y, CHEN M, et al. Studies on the pollution levels of antibiotic resistance genes in Jiulong River estuary and wastewater treatment plants in Xiamen [J]. Microbial Bulletin, 2012, 39(5): 683−695.(in Chinese)
[7]	ZHU Y G, ZHAO Y, LI B, et al. Continental-scale pollution of estuaries with antibiotic resistance genes [J]. Nature Microbiology, 2017(2): 16270.
[8]	刘中柱, 郑伟文. 中国满江红[M]. 北京: 农业出版社, 1989.
[9]	BECKING J H. Endophyte transmission and activity in the Anabaena-Azolla association [J]. Plant and Soil, 1987, 100(1/2/3): 183−212.
[10]	PETERS G A, MEEKS J C. The Azolla-Anabaena symbiosis: Basic biology [J]. Annual Review Plant Physiology and Plant Molecular Biology, 1989, 40: 193−210. doi: 10.1146/annurev.pp.40.060189.001205
[11]	ITO O, WATANABE T. The relationship between combined nitrogen uptaken and nitrogen fixation in Azolla-Anabaena symbioses [J]. New Phytologist, 1983, 95: 647−654. doi: 10.1111/j.1469-8137.1983.tb03528.x
[12]	GATES J E, FISHER R W, CANDLER R A. The occurrence of coryneform bacteria in the leaf cavity of Azolla [J]. Archives of Microbiology, 1980, 127(2): 163−165. doi: 10.1007/BF00428020
[13]	SERRANO R, CARRAPICO F, VIDAL R. The presence of lectins in bacteria associated with Azolla-Anabaena symbiosis [J]. Symbiosis, 1999, 15: 169−178.
[14]	LECHNO Y S, NIERZWICKI-BAUER S A, RAI A N, et al. Cyanobacteria in symbiosis[M]. Dordrechr. The Netherlands: Kluwer Academic Publishers, 2002: 153-178.
[15]	ZHENG S P, CHEN B, GUAN X, et al. Diversity analysis of endophytic bacteria within Azolla microphyllausing PCRGGE and electron microscopy [J]. Chinese Journal of Agricultural Biotechnology, 2008, 5(3): 269−276. doi: 10.1017/S1479236208002441
[16]	郑斯平, 陈彬, 王瑾, 等. 小叶满江红(Azolla microphylla)内生细菌多样性的T-RFLP分析 [J]. 安徽农业科学, 2012, 40(29):14185−14187, 14270. doi: 10.3969/j.issn.0517-6611.2012.29.014 ZHENG S P, CHEN B, WANG J, et al. T-RFLP Analysis on diversity of endophytic bacteria in Azolla microphylla [J]. Journal of Anhui Agriculture Sciences, 2012, 40(29): 14185−14187, 14270.(in Chinese) doi: 10.3969/j.issn.0517-6611.2012.29.014
[17]	SU J Q, AN X L, LI B, et al. Metagenomics of urban sewage identifies an extensively shared antibiotic resistome in China [J]. Microbiome, 2017, 5(1): 1−15. doi: 10.1186/s40168-016-0209-7
[18]	彭司华, 吴智超, 孙丹, 等. 宏基因组学在抗生素抗性基因鉴定中的应用 [J]. 基因组学与应用生物学, 2019, 38(9):4102−4109. PENG S H, WU Z C, SUN D, et al. Application of Metagenomics in the Identification of Antibiotic Resistance Genes [J]. Genomics and Applied Biology, 2019, 38(9): 4102−4109.(in Chinese)
[19]	白克智, 于赛玲, 陈维纶, 等. 无藻满江红和满江红鱼腥藻的分离与培养 [J]. 科学通报, 1979, 14:664−666. BAI K Z, YU S L, CHEN W L, et al. Isolation and pure culture of algae-free Azollaand Anabaena azollae [J]. Chinese Sciences Bulletin, 1979, 14: 664−666.(in Chinese)
[20]	BÜNGER W, JIANG X, MÜLLER J, et al. Novel cultivated endophytic Verrucomicrobia reveal deep-rooting traits of bacteria to associate with plants [J]. Scientific Reports, 2020, 10(1): 8692. doi: 10.1038/s41598-020-65277-6
[21]	LIU B, POP M. ARDB: antibiotic resistance genes database [J]. Nucleic Acids Research, 2009, 37(Database issue): D443−D447.
[22]	JIA B F, RAPHENYA A R, ALCOCK B, et al. CARD 2017: expansion and model-centric curation of the comprehensive antibiotic resistance database [J]. Nucleic Acids Research, 2016: gkw1004.
[23]	曾秀丽, 王志, 罗利, 等. 茶树内生草螺菌ZXN111生长素合成及其对云抗-10号植物的促生功能 [J]. 微生物学报, 2020, 60(10):2198−2210. ZENG X L, WANG Z, LUO L, et al. Auxin synthesis in tea plant endophytic Herbaspirillum sp. and the plant growth promotion on Yunkang-10 [J]. Acta Microbiologica Sinica, 2020, 60(10): 2198−2210.(in Chinese)
[24]	沈应博, 史晓敏, 沈建忠, 等. 全基因组测序与生物信息学分析在细菌耐药性研究中的应用 [J]. 生物工程学报, 2019, 35(4):541−557. SHEN Y B, SHI X M, SHEN J Z, et al. Application of whole genome sequencing technology and bioinformatics analysis in antimicrobial resistance researches [J]. Chinese Journal of Biotechnology, 2019, 35(4): 541−557.(in Chinese)
[25]	杨兵, 梁晶, 刘林梦, 等. 耐药基因数据库概述 [J]. 生物工程学报, 2020, 36(12):2582−2597. YANG B, LIANG J, LIU L M, et al. Overview of antibiotic resistance genes database [J]. Chinese Journal of Biotechnology, 2020, 36(12): 2582−2597.(in Chinese)
[26]	MODI S R, LEE H H, SPINA C S, et al. Antibiotic treatment expands the resistance reservoir and ecological network of the phage metagenome [J]. Nature, 2013, 499(7457): 219−222. doi: 10.1038/nature12212
[27]	NESME J, CECILLON S, DELMONT T O, et al. Large-scale metagenomic-based study of antibiotic resistance in the environment [J]. Current Biology, 2014, 24(10): 1096−1100. doi: 10.1016/j.cub.2014.03.036
[28]	谭文彬. 细菌耐药的基因机理研究进展 [J]. 中国病原生物学杂志, 2009, 4(7):543−544, 555. TAN W B. Research advances in genetic mechanisms of bacterial antibiotic resistance [J]. Journal of Pathogen Biology, 2009, 4(7): 543−544, 555.(in Chinese)
[29]	朱阵, 曹明泽, 张吉丽, 等. 细菌耐药性研究进展 [J]. 中国畜牧兽医, 2015, 42(12):3371−3376. ZHU Z, CAO M Z, ZHANG J L, et al. Research progress on bacterial resistance [J]. China Animal Husbandry and Veterinary Medicine, 2015, 42(12): 3371−3376.(in Chinese)