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科技导报 |研究论文 2012 , 30 (27) : 20 -23
一株拮抗芽孢杆菌的分离鉴定及其β-1,4-葡聚糖酶基因的克隆
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吴慧玲, 刘伟成, 卢彩鸽, 董丹, 张殿鹏, 张涛涛
作者信息
    北京市农林科学院植物保护环境保护研究所,北京 100097
通讯作者:
刘伟成,研究员,研究方向为病害生物防治,电子信箱:liuwc@126.com
Isolation and Identification of an Antagonistic Bacillus strain and Its β-1,4-Glucanase Gene Cloning
  • WU Huiling, LIU Weicheng, LU Caige, DONG Dan, ZHANG Dianpeng, ZHANG Taotao
  • Affiliations
      Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
    出版时间: 2012-09-28 doi: 10.3981/j.issn.1000-7857.2012.27.001
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    植物真菌病害是影响农业生产的主要因素之一,连作栽培导致病害逐年加重,为获得能够抑制病原真菌的生防菌以解决真菌病害防治难题,通过平板对峙实验及分子鉴定、抑菌活性物质分析、葡聚糖酶基因克隆及分析,获得一株对多种植物病原真菌具有显著抑制作用的芽孢杆菌,菌株编号为L103。16S rDNA及看家基因序列分析表明,该菌株为巨大芽孢杆菌,羧甲基纤维素钠平板筛选及刚果红染色实验发现该菌具有很强的产纤维素酶(CMC酶)的能力,根据GenBank中巨大芽孢杆菌内切葡聚糖酶基因(HM130670.1)序列设计引物,将扩增得到的1482bp大小的片段克隆到载体pMD18-T上,测序并通过GenBank数据库BLAST分析显示该序列与一株Bacillus sp. HB102的内切β-1,4-葡聚糖酶基因序列一致性达到95%,而与Bacillus subtilisβ-1,4-内切葡聚糖酶基因一致性为85%,进一步分析该菌株产生的抗病活性物质并对其进行改良将为该菌在今后的真菌病害防治应用中提供理论基础。
    巨大芽孢杆菌  /  16S rDNA  /  内切β-1  /  4-葡聚糖酶
    Plant fungal diseases are the main factor that affects agricultural production. In order to inhibit fungus diseases, the flat confrontation experiment and the molecular detection were employed for the isolation of biocontrol bacteria. Bacillus L103 was isolated from a vegetable field in suburb of Beijing. The antimicrobial spectrum suggests that this strain shows a high level of inhibitory activity against the tested plant pathogenic fungi. The analysis of 16S rDNA and housekeeping gene sequence shows that it is a kind of Bacillus megaterium. It could produce β-1,4-glucanase, that was detected on CMC-Na plates and congo red staining. We amplified the endo-β-1,4-glucanase gene and sequenced it according to GenBank No.HM130670.1. Its full length was found to be 1482bp and its amino acid number is about 494. Its endo-β-1,4-glucanase gene sequence has 92% homology with that of the endo-β-1,4-glucanase gene of Bacillus sp. HB102, and 85% homology with that of Bacillus subtilis.
    Bacillus megaterium  /  16S rDNA  /  β-1  /  4-glucanase
    吴慧玲;刘伟成;卢彩鸽;董丹;张殿鹏;张涛涛. 一株拮抗芽孢杆菌的分离鉴定及其β-1,4-葡聚糖酶基因的克隆. 科技导报, 2012 , 30 (27) : 20 -23 . DOI: 10.3981/j.issn.1000-7857.2012.27.001
    WU Huiling;LIU Weicheng;LU Caige;DONG Dan;ZHANG Dianpeng;ZHANG Taotao. Isolation and Identification of an Antagonistic Bacillus strain and Its β-1,4-Glucanase Gene Cloning[J]. Science & Technology Review, 2012 , 30 (27) : 20 -23 . DOI: 10.3981/j.issn.1000-7857.2012.27.001

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    doi: 10.3981/j.issn.1000-7857.2012.27.001
    • 接收时间:2012-06-28
    • 首发时间:2012-09-28
    • 出版时间:2012-09-28
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    • 收稿日期:2012-06-28
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    刘伟成,研究员,研究方向为病害生物防治,电子信箱:liuwc@126.com
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    2种不同金属材料的力学参数

    Family
    属数
    Number of
    genus
    种数
    Number of
    species
    占总种数比例
    Percentage of
    total species (%)

    Genus
    种数
    Number of
    species
    占总种数比例
    Percentage of total
    species (%)
    鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
    小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
    多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
    红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
    小菇属 Mycena 11 5.26
    光柄菇属 Pluteus 5 2.39
    红菇属 Russula 17 8.13
    栓菌属 Trametes 5 2.39
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