Diversity and Mercury-resistance of Actinomycetes in Tongren, Guizhou
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摘要: 为探明贵州喀斯特重金属污染区域土壤放线菌多样性特征以及为其污染修复应用奠定基础，从贵州铜仁地区的25份土壤样品中，采用梯度稀释涂布法，利用3种处理方式、7种培养基分离放线菌；通过形态特征、16S rDNA基因序列分析结合生理生化特征鉴定放线菌；同时筛选重金属汞抗性菌株。结果从土样中共分离出56株典型放线菌菌株，经初步鉴定，分属于链霉菌属、孢囊链霉菌属、高温单孢菌属、线杆菌属、间孢囊菌属、诺卡氏菌属、小单孢菌属；其中链霉菌属放线菌占68%。筛选出了1株对汞有较高抗性（75 mg·L-1）的放线菌菌株，优良耐受菌株的最大抗性约85 mg·L-1，菌株经鉴定为枝链霉菌Streptomyces rameus。共分离出56株，分属于7个属的放线菌菌株；筛选出1株重金属汞高抗菌株，经鉴定为枝链霉菌。Abstract: Diversity and mercury-resistance of Actinomycetes in the soil polluted by heavymetals at Tongren karst regeions in Guizhou were studied to gain information for environmental recovery and bioremedies. Twenty-five soil samples were collected from the areas. Microorganisms were screened on 7 different culture media with gradient concentrations of mercury under 3 different treatments to isolate the resistant strains. Subsequently, the isolates were examined for their morphological, physiological and biological characteristics, and subjected to a 16S rDNA phylogenetic an alysis for further identification.As a result, 56 typical Actinomycetes belonging to Streptomyces, Streptosporangium, Thermomonospora, Actinobacillus, Intrasporangium, Nocardia, and Micromonospora were found to be resistant to mercury.Among them, 68% were Streptomyces with one particular strain showing a high mercury-resistance to survive under a 75 mg·L-1 up to 85 mg·L-1 mercury stress, which was identified to be Streptomyces rameus.
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Key words:
- Actinomycetes /
- diversity /
- karst /
- 16S rDNA /
- mercury /
- Streptomyces rameus
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图 3 菌株WS1605的PCR产物电泳
注：M为Marker；1为WS1605。
Figure 3. Electrophoresis of PCR amplification of WS1605
图 4 基于WS1605 16S rDNA构建的系统发育树
Figure 4. Phylogenetic trees based on 16S rDNA of WS1605 and related strains
表 1 重铬酸钾对放线菌分离的影响
Table 1. Effect of potassium dichromate on separating strains of Actinomycetes
培养基 重铬酸钾 对照组 总菌落数/×105 放线菌数/×105 杂菌率/% 总菌落数/×105 放线菌数/×105 杂菌率/% 高氏1号 1.2 1.12 6.8 0.7 0.51 27.5 LNMS 3.1 2.84 8.5 3.5 1.23 65 
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表 2 不同分离培养基对放线菌影响
Table 2. Effect of culture media on separating strains of Actinomycetes
培养基种类 放线菌数量/×105 放线菌种类 种类数 高氏1号培养基 1.35 孢囊链霉菌属；灰红紫类群；灰褐类群；粉红孢类群；黄色类群；烬灰类群 6 LNMS培养基 5.2 高温单孢菌属；烬灰类群；粉红孢类群；金色类群 4 海藻糖-脯氨酸培养基 0.95 线杆菌属；粉红孢类群；金色类群；烬灰类群 4 改良高氏二号培养基 1.1 线杆菌属；间孢囊菌属；灰褐类群；粉红孢类群 4 改良脯氨酸培养基 2.7 粉红孢类群；小单孢菌属；金色类群 3 ZSSE培养基 2.1 诺卡氏菌属；小单孢菌属；金色类群 3 NA浸出液培养基 0.76 黄色类群；灰红紫类群；灰褐类群；金色类群；烬灰类群；小单孢菌属 6
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表 3 菌株WS1605生理生化特征
Table 3. Physiological and biochemical characteristics of WS1605
项目 结果 柠檬酸盐试验 - 硝酸盐还原 - 淀粉水解 + 牛奶分解 + 明胶液化 - 蔗糖 + 果糖 + 肌醇 + 鼠李糖 + 山梨醇 - 柠檬酸钠 - 酪氨酸酶 + L-阿拉伯糖 - 半乳糖 + 注：“+”表示阳性；“－”表示阴性。
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