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高耐镉菌株PDA-H的筛选及活性研究 摘 要 当下，土壤重金属污染不断加重，在我国，土壤镉（下文简称Cd）污染尤为显著，造成了许多危害。重金属污染土壤的微生物修复作为一种低成本、操作方便、应用前景广泛的绿色修复技术，倍受国内外研究者关注。本文以从苏州地区典型污染场地土壤与高有机质土壤中筛选出的耐Cd菌株PDA-H为研究对象，进行菌株复筛、耐Cd性能测定、菌种鉴定及单一介质中基础条件下其活性及Cd吸附情况研究。实验结果表明，菌株PDA-H具有很强的耐Cd能力，适宜在马铃薯（PDA）培养基中生长，不适宜在葡萄糖-蛋白胨培养基中生长，其在PDA固体培养基中能耐受的Cd浓度最高达1.4 g/L，在PDA液体培养基中能耐受的Cd浓度最高达0.8 g/L。生理生化鉴定及26S rDNA基因序列鉴定表明，菌株PDA-H为皱纹表膜假丝酵母（Candida rugopel -liculosa strain NRRL Y-17079）。对Cd胁迫下PDA-H在菌液中的生物量、蛋白含量、DNA含量、淀粉酶活力、脂肪酶活力及其Cd吸附能力随时间的变化进行考察，发现其分泌的淀粉酶及脂肪酶对Cd污染表现均很敏感，其中淀粉酶较脂肪酶更为敏感，两种酶活性均有望作为生物指标应用于Cd污染的监测中。对菌株Cd吸附能力的考察表明随时间推移，PDA-H对Cd的吸附率总体呈不断增加趋势，最大吸附率达近60%。此外，Cd吸附率与脂肪酶活性呈较好的正相关关系，与淀粉酶活性呈较好的负相关关系。菌株PDA-H有望在Cd污染场地的微生物修复工程中得到应用。 Screening and activity studies of PDA-H with high cadmium resistance Abstract At present, the heavy metal pollution of the soil is increasing. In China, soil cadmium ( Cd ) pollution is particularly remarkable, causing many hazards. Microbial remediation of heavy metal contaminated soil is a green repair technology with low cost, convenient operation and wide application prospect, many researchers all around the world pay more attention to it. This paper use a Cd resistant strain ( named PDA-H ) which was selected from soil in SuZhou typical contaminated and high organic matter site as the study object, carry out the second screening, Cd resistance determination, identification, the activity and adsorption of this strain in the single medium under the basic conditions. From these experiements, we can know PDA-H has strong resistance to Cd. This strain enjoy growing in Potato( PDA ) medium but is not suitable to live in glucose peptone mesium. The highest Cd concentration PDA-H can tolerate in solid PDA medium is 1.4 g/L, in liquid PDA medium the concentration is 0.8 g/L. From the 26S rDNA gene sequence identification, combine with physiological and biochemical identification, we can determine PDA-H is Candida rugopel -liculosa strain NRRL Y-17079. Investigating the biomass, protein content, DNA content, amylase activity, lipase activity and Cd adsorption ability of PDA-H in bacteria solution with Cd stress, it is found that amylase and lipase from PDA-H are all sensitive to Cd pollution, amylase was more sensitive than lipase. The two enzymes are expected to be used as biological indicators for the monitoring of Cd pollution. The adsorption capacity of Cd of this strain showed that the adsorption rate was increasing with the passage of time in general, and the maximum adsorption rate was close to 60%. The adsorption rate of Cd and lipase activity present positive correlation roughly, but it was just the opposite with lipase activity. Strain PDA-H is expected to be applied in microbial remediation of Cd contaminated sites. |
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2楼2015-06-10 23:33:38