| 李一平，沈汉国，喻国辉，王 燕，李 驰，叶志文，周东辉，吴绪波.一种芽胞杆菌生物被膜形成缺陷培养基的研究[J].广东农业科学,2016,43(9):98-104 |
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一种芽胞杆菌生物被膜形成缺陷培养基的研究 |
| Study on a bacillus biofilm growth defective culture medium |
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| DOI：10.16968/j.issn.1004-894X.2016.09.015 |
| 中文关键词: 枯草芽胞杆菌 生物被膜 培养基 菌落 薄皮 |
| 英文关键词: Bacillus subtilis biofilm culture medium colony pecille |
| 基金项目:广东省科技计划项目（2014A020208015） |
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| 中文摘要: |
| 为寻找一种不支持芽胞杆菌正常形成生物被膜的培养基，比较枯草芽胞杆菌R31 和168 菌株
在MSgg、NB、LB、BGM 固体和液体培养基中形成的生物被膜形态差异，确定R31 和168 菌株生物被膜形
成能力差异；根据R31 在MSgg、NB、BGM 培养基中振荡培养的生长方式设计出BGM1 和BGDM 培养基，
测定R31 在BGM1 和BGDM 培养基上的生长曲线并比较R31 菌株和168 菌株在BGM1 和BGDM 上生物
被膜形成的差异，然后测定枯草芽胞杆菌TR21 菌株、解淀粉芽胞杆菌R21g9 菌株、短芽胞杆菌L1 菌株、
胶质芽胞杆菌L2 菌株在BGM1 和BGDM 培养基上生物被膜形成情况。结果显示，R31 在MSgg、NB、LB、
BGM 液体培养基表面形成薄皮，在MSgg、LB 固体培养基和NA 上形成具有褶皱的菌落，在BGM 上形成光
滑的菌落，R31 在这些培养基中形成的生物被膜比168 菌株厚实。R31 在MSgg 和BGM 中早期（2 h）以游
离状态生长，对数生长期（4 h）菌体黏连出现链状，对数生长中后期（8 h）菌体呈现网状结构，但在NB
培养基中主要以游离状态生长，显示营养而非培养条件影响R31 的生长方式。于是将BGM 培养基中的酵
母提取物换为牛肉膏获得BGM1 培养基，将BGM1 培养基的胰蛋白胨改为细菌学蛋白胨得到BGDM 培养
基，并发现R31 和168 菌株可以在BGM1 上形成薄皮和菌落，不能在BGDM 培养基上形成完整的薄皮和具
有褶皱的菌落。待测菌株均能够在BGM1 培养基上形成厚薄程度有差异的薄皮和有褶皱的菌落，均不能在
BGDM 培养基上形成薄皮和具有褶皱的菌落。在BGDM 中添加甘油或Mn 可以恢复R31 形成薄皮和扩散的
褶皱菌落。芽胞杆菌在BGDM 培养基上不能正常形成薄皮和具有褶皱的菌落，BGDM 可以用于筛选促进生
物被膜形成的物质。 |
| 英文摘要: |
| Abstract：To find a culture medium could not support bacillus biofilm formation，the colony and pellicle
morphological differences of Bacillus subtilis stain R31 and 168 in solid and liquid MSgg，NB，LB and BGM medium
were compared to make sure the biofilm formation difference of the two strains. Based on the different growth process
of R31 in liquid MSgg，NB and BGM on shaking cultivation，two culture mediums were designed and named BGM1
and BGDM. The growth curve and process of R31 in BGM1 and BGDM were checked and the colony and pellicle
morphological differences of R31 and 168 developed in BGM1 and BGDM were compared. Finally，the biofilm
morphological differences of B. subtilis strain TR21，B. amyloliquefaciens strain R21g9，Brevibacillus sp. strain L1
and B. mucilaginosus strain L2 in BGM1 and BGDM were studied. Results showed that R31 could develop integrated
pellicle in liquid MSgg，NB，LB and BGM medium，and develop wrinkle colony in solid MSgg，LB and NA medium，but develop smooth colony in solid BGM medium. The biofilm of R31 in these mediums were more robust
than that of 168. The cells of R31 grew freely in MSgg and BGM medium in the early stage (2 h) of cultivation，the
cells attached to each other and linked like a chain in logarithmic phase (4 h)，and the cell morphology looked like a
net in the end of logarithmic phase (8 h)，but R31 cells grew freely in NB culture medium at the whole growth process.
These results showed that nutrition but not cultural condition affected the growth process of R31. So the yeast extract
in BGM was changed as beef extract and the medium named BGM1，and tryptone of BGM1 was changed as peptone
and the medium was named BGDM. R31 and 168 could develop pellicle and colony in BGM，but all couldn’t
develop pellicle and winkle colony in BGDM medium. All the tested bacillus strains could develop integrated pellicle
with different thickness and wrinkle colony in BGM1，but they all could not develop integrated pellicle and wrinkle
colony in BGDM medium. Glycerol or Manganese were added in BGDM could recover the integrated pellicle and
wrinkle spread colony formation of R31. Bacillus strains could not develop normal pellicle and winkle colony in
BGDM medium，and this medium could be used to select the materials. |
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