生物技术通报 ›› 2022, Vol. 38 ›› Issue (9): 136-146.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0112

• 细菌耐药性专题(专题主编: 刘雅红 教授 孙坚 教授) • 上一篇    下一篇

无色杆菌77的基因组构成及其趋化和耐药特性

李霁虹1,2(), 荆玉玲2, 马桂珍1(), 郭荣君2(), 李世东2   

  1. 1.江苏海洋大学环境与化学工程学院,连云港 222005
    2.中国农业科学院植物保护研究所,北京 100193
  • 收稿日期:2022-01-25 出版日期:2022-09-26 发布日期:2022-10-11
  • 作者简介:李霁虹,女,硕士研究生,研究方向:生物化工;E-mail: 804797175@qq.com
  • 基金资助:
    国家重点研发计划(2019YFD1002000);现代农业产业技术体系项目(CARS-23-C04)

Genome Construction of Achromobacter 77 and Its Characteristics on Chemotaxis and Antibiotic Resistance

LI Ji-hong1,2(), JING Yu-ling2, MA Gui-zhen1(), GUO Rong-jun2(), LI Shi-dong2   

  1. 1. College of Environmental and Chemical Engineering,Jiangsu Ocean University,Lianyungang 222005
    2. Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193
  • Received:2022-01-25 Published:2022-09-26 Online:2022-10-11

摘要:

无色杆菌77(Achromobacter sp. 77)是黄瓜枯萎病菌(Fusarium oxysporum f. sp. cucumerinum,Foc)菌丝际优势细菌,可随Foc菌丝迁移且状态活跃。为明确菌株77在Foc菌丝际的定殖机理,本研究在分析菌株77全基因组构成基础上,研究了其趋化特性和耐药性。Illumina Hiseq+PacBio测序分析表明,菌株77基因组全长5 868 070 bp,仅含1条染色体,GC含量为65.89%。在KEGG、COG和GO数据库分别注释到2 696、4 862和4 212个基因。菌株77基因组中含mcpchemot等重要趋化基因;通过CARD数据库注释,发现含有drremr等抗生素耐药外排泵基因、抗生素耐药基因簇以及多种抗生素的抗性基因。抗生素敏感性实验表明,菌株77对浓度低于300 µg/mL的氯霉素、卡那霉素、氨苄青霉素以及浓度低于50 µg/mL的四环素均有抗性,是一株多重耐药菌株。采用改良的游动平板法测定菌株77的趋化特性,结果表明,菌株77对1 mmol/L对羟基苯乙酸、水杨酸、α-酮戊二酸、延胡索酸、琥珀酸、苹果酸等多种根分泌物和真菌分泌物组分具有明显的趋化响应;0.5 mmol/L的上述化合物和0.5-1 mmol/L的镰刀菌酸可促进菌株77的生长。上述研究结果说明菌株77对根分泌物或真菌分泌物具有趋化和利用能力,是连作条件下无色杆菌属细菌在枯萎病菌菌丝际丰度升高的重要原因。

关键词: 无色杆菌, 黄瓜枯萎病菌, 全基因组分析, 趋化, 耐药

Abstract:

Achromobacter sp. 77,the dominant hyphosphere bacterium of Fusarium oxysporum f. sp. cucumerinumFoc),actively migrates along with Foc hyphae growth. To clarify the colonization mechanism of strain 77 in Foc hyphosphere,the whole genome of strain 77 was sequenced by using Illumina Hiseq+PacBio sequencing method,and its characteristics on chemotaxis and antibiotic resistance were examined. The genome of strain 77 was 5 868 070 bp with one chromosome and GC content of 65.89%. Total of 2 696,4 862 and 4 212 genes were annotated against KEGG,COG and GO database,respectively. Gene annotation demonstrated that strain 77 contained multiple important chemotactic genes such as mcp,che and mot in its genome. Annotation of its genome against CARD database showed that it harbored antibiotic resistance efflux pump genes such as drr,emr and gene clusters resistant to multiple antibiotics. Antibiotic sensitivity tests showed that strain 77 was resistant to chloramphenicol,kanamycin and ampicillin at the concentration < 300 µg/mL,and resistant to tetracycline at the concentration < 50 µg/mL,indicating that strain 77 was a multi-drug resistant strain. The chemotactic test indicated that strain 77 showed obviously chemotactic response to 1 mmol/L p-hydroxyphenylacetic acid,salicylic acid,α-ketoglutaric acid,fumaric acid,succinic acid and malic acid. Moreover,the growth of strain 77 was promoted by the above compounds at the concentration of 0.5 mmol/L and by 0.5-1.0 mmol/L of fusaric acid. These results indicated that strain 77 showed chemotaxis toward root or fungal exudates and utilization them,which might be the reason that the abundance of Achromobacter increased in Foc hyphosphere after continuous cultivation of cucumber in Foc infested soil.

Key words: Achromobacter, Fusarium oxysporum f. sp. cucumerinum, whole genome sequencing, chemotaxis, antibiotic resistance