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氧化乐果降解菌ZZY-C13-1-9的筛选、鉴定及降解能力

陈锐 瞿佳 门欣 邓媛 孙晓宇 赵玲侠 沈卫荣

陈锐,瞿佳,门欣,等. 氧化乐果降解菌ZZY-C13-1-9的筛选、鉴定及降解能力 [J]. 福建农业学报,2020,35(8):869−875 doi: 10.19303/j.issn.1008-0384.2020.08.009
引用本文: 陈锐,瞿佳,门欣,等. 氧化乐果降解菌ZZY-C13-1-9的筛选、鉴定及降解能力 [J]. 福建农业学报,2020,35(8):869−875 doi: 10.19303/j.issn.1008-0384.2020.08.009
CHEN R, QU J, MEN X, et al. Isolation, Identification, and Characterization of Omethoate-degrading Pseudomonas abietaniphila ZZY-C13-1-9 [J]. Fujian Journal of Agricultural Sciences,2020,35(8):869−875 doi: 10.19303/j.issn.1008-0384.2020.08.009
Citation: CHEN R, QU J, MEN X, et al. Isolation, Identification, and Characterization of Omethoate-degrading Pseudomonas abietaniphila ZZY-C13-1-9 [J]. Fujian Journal of Agricultural Sciences,2020,35(8):869−875 doi: 10.19303/j.issn.1008-0384.2020.08.009

氧化乐果降解菌ZZY-C13-1-9的筛选、鉴定及降解能力

doi: 10.19303/j.issn.1008-0384.2020.08.009
基金项目: 陕西省科学院科技计划项目(2018K-08、2018NK-01)
详细信息
    作者简介:

    陈锐(1977−),女,博士,副研究员,研究方向:微生物资源、污染环境生物修复(E-mail:email4rui@163.com

  • 中图分类号: Q 939.96

Isolation, Identification, and Characterization of Omethoate-degrading Pseudomonas abietaniphila ZZY-C13-1-9

  • 摘要:   目的  从设施棚室土壤中筛选获得可降解氧化乐果的微生物菌株,研究降解菌对氧化乐果在土壤中残留量的降解效果。  方法  通过唯一碳源筛选法筛选土壤样本中的本源菌株,生理生化及16S rDNA鉴定种属类型,通过钼胺蓝显色法及HPLC法验证菌株对氧化乐果的降解效果。在土壤中添加一定量的氧化乐果,再添加微生物菌剂并保持湿度温度以验证微生物菌剂在土壤中的应用效果。  结果  筛选获得一株可分解利用氧化乐果的微生物菌株ZZY-C13-1-9,该菌株可在含1 000 mg·L−1的氧化乐果无机盐培养基中正常生长。经鉴定确定该菌为嗜松香假单胞菌(Pseudomonas abietaniphila)。经HPLC证实,在含氧化乐果400 mg·L−1发酵培养基中,30 ℃、180 r·min−1摇瓶培养240 h,氧化乐果的降解率达92.3%。ZZY-C13-1-9亦可在敌百虫与敌敌畏中正常生长。土壤室内试验证明:土壤水分含量保持25%~30%,室内温度25~34 ℃,45 d后嗜松香假单胞菌ZZY-C13-1-9可将土壤中200 mg·L−1的氧化乐果降解59.2%。  结论  筛选获得一株对土壤中残留的氧化乐果具有降解作用的菌株。
  • 图  1  氧化乐果残留含量

    注:**表示不同处理间差异达极显著水平(P<0.01),图4同。

    Figure  1.  Omethoate residue in fermentation broth

    Note: **indicates significantly different at 0.01 level. Same for Fig. 4.

    图  2  ZZY-C13-1-9菌株形态鉴定

    注:a.菌落形态;b.革兰氏染色。

    Figure  2.  Morphology of ZZY-C13-1-9

    Note:a. colony morphology; b. gram stain.

    图  3  ZZY-C13-1-9 与模式菌株系统进化树

    Figure  3.  Phylogenetic tree of ZZY-C13-1-9

    图  4  氧化乐果残留量

    Figure  4.  Quantity of omethoate residues

    表  1  土壤样本信息

    Table  1.   Information on soil sample

    样本编号
    Sample ID
    样本来源
    Source
    经度
    Longitude(E)
    纬度
    Latitude(N)
    海拔
    Altitude/m
    土壤类型
    Soil type
    植被
    Plant
    TY21陕西省西安市周至县田峪108.20.38°34.3.20°660壤土 Loam核桃树 Walnut
    TY 44陕西省西安市周至县田峪108.20.38°34.3.20°660壤土 Loam柿子树 Persimmon
    QL107陕西省宝鸡市太白县太白山107.47.52°33.59.18°3510壤土 Loam猕猴桃 Kiwi
    QL106陕西省宝鸡市太白县太白山107.47.53°33.59.25°3495壤土 Loam猕猴桃 Kiwi
    BF19陕西省西安市周至县厚畛子镇花耳坪村107.49.79°33.49.75°1317壤土 Loam细辛 Asarum
    BF12陕西省西安市周至县厚畛子镇黑河口107.49.72°33.49.75°1312壤土 Loam七叶一枝花 Paris polyphylla
    ZZ34陕西省西安市周至县尚村镇涧里村107.31°34.05°1543壤土 Loam枣树 Jujube
    ZZ37陕西省西安市周至县终南镇毓兴村107.31°34.05°1543壤土 Loam枣树 Jujube
    WN1-2陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN1-1陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN1-3陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN1-4陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN2-1陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN2-2陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN2-3陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    WN2-4陕西省渭南市大荔县埝桥乡东埝村109.87°34.82°354壤土 Loam黄瓜 Cucumber
    T-13陕西省西安市周至县楼观镇羊坡村107.31°34.05°1546壤土 Loam枣树 Jujube
    T-20陕西省西安市周至县马召镇郭家寨107.31°34.05°1527壤土 Loam枣树 Jujube
    T-24陕西省西安市周至县竹峪镇兰塬村107.31°34.05°1532壤土 Loam枣树 Jujube
    T-45陕西省西安市周至县楼观镇下三清寸107.31°34.05°1555壤土 Loam枣树 Jujube
    T-46陕西省西安市周至县107.31°34.05°1528壤土 Loam枣树 Jujube
    T-51陕西省西安市周至县集贤镇大曲村107.31°34.05°1574壤土 Loam枣树 Jujube
    T-64陕西省西安市周至县107.31°34.05°1544壤土 Loam枣树 Jujube
    T-67陕西省西安市周至县107.31°34.05°1543壤土 Loam枣树 Jujube
    下载: 导出CSV

    表  2  ZZY-C13-1-9对其他有机磷农药的耐受

    Table  2.   Tolerance of ZZY-C13-1-9 to organophosphorus pesticides

    测定值 Values农药 Pesticide质量浓度 Concentration/(mg·L−1
    1000 500100 0
    OD600nm敌百虫 Dipterex0.45±0.050.40±0.010.50±0.050.45±0.01
    敌敌畏 DDVP0.73±0.400.59±0.090.75±0.03-
    下载: 导出CSV

    表  3  土壤室内试验氧化乐果残留量及降解率(t-test,P=0.016)

    Table  3.   Residues and degradation rate of omethoate in laboratory soil (t-test, P=0.016)

    处理
    Treatment
    残留量
    Residual quantity/(mg·L−1
    降解率
    Degradation rate/%
    0 d45 d
    对照 CK 224.10 169.58 24.33 B
    ZZY-C13-1-9 213.42 87.07 59.20 A
    注:不同大写字母表示差异极显著(P<0.01)。
    Note: Data with different uppercase letters indicate extremely significant difference (P<0.01).
    下载: 导出CSV
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  • 收稿日期:  2020-02-12
  • 修回日期:  2020-04-28
  • 刊出日期:  2020-08-19

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