不同矿物对氧化亚铁硫杆菌YN-3中磁小体形成的影响
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摘要
趋磁细菌(Magnetotactic bacterium)是对体内含有磁性颗粒并表现出趋磁性的所有细菌的通称。在其体内含有的磁性颗粒被称为—磁小体。磁小体具有粒度小而均匀,不团聚,纯度高,结晶完美,磁性质独特,无细胞毒性等特点,是一种理想的生物纳米材料。
本课题组发现在氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)等浸矿菌中含有生物控制矿化作用所产生的磁小体。本文主要研究不同矿物作用对浸矿细菌中磁小体形成的影响。
本文以氧化亚铁硫杆菌菌株YN-3为材料,研究在黄铁矿、黄铜矿、磁黄铁矿和闪锌矿等矿物作用下,对YN-3的生长特性及趋磁性产生的影响。结果表明,在不同矿物的作用下,细菌的生长特性有很大不同。在以闪锌矿为能源时,菌体生长速率最快,在第11天,菌量达到最大值2.73X107。其次是磁黄铁矿和黄铜矿,而黄铁矿作用时菌体生长速率最慢。半固体滋泳表明,以黄铁矿和磁黄铁矿作为能源培养的细菌会表现出趋磁性,而利用黄铜矿与铁闪锌矿时无此现象。
利用实时定量PCR技术研究YN-3中与磁小体形成相关的mpsA、magA、thy和mamB基因分别在黄铁矿、黄铜矿、磁黄铁矿和闪锌矿的作用下的表达差异。实时定量PCR结果表明,在以磁黄铁矿和黄铁矿为能源时,四个基因表达量均高于其它两种矿。但是以黄铁矿为能源时,由于矿体难以氧化,菌体生长极为缓慢,难以大量培养收集。综合趋磁性和细菌生长特性两方面因素可知,在磁黄铁矿作用下,YN-3形成磁小体的效果最好,磁黄铁矿为最佳矿物能源。
本文利用不同浓度的磁黄铁矿作用于YN-3,研究不同浓度磁黄铁矿对YN-3的生长状况及对其形成磁小体的影响。实验中磁黄铁矿浓度分别1%、2%、3%、4%、5%。生长曲线结果表明,磁黄铁矿浓度为3%时,菌体生长速度最快。浓度为5%时,菌体生长极为缓慢。实时定量PCR结果表明mpsA, magA和mamB基因随着磁黄铁矿浓度的提高,表达量呈增大趋势。磁黄铁矿浓度为1%-3%时,表达量增加明显。超过3%时,增长趋于平缓。Thy基因在各浓度磁黄铁矿作用下,表达量均较低,没有明显变化。综合生长曲线和实时定量PCR结果两个因素表明,3%为磁黄铁矿对YN-3作用的最适浓度。
Magnetotactic bacteria(MTB) is the name of all kinds of prokaryotes with magnetotaxis, which containing Nano-particles--the magnetosomes. Magnetosomes is an ideal kind of bio-nano-magnetic material, because of its specific properties such as superior crystalline, well biocompatible, uniform nano-sized, non-agglomerated and specific magnetic characteristics.
The researchers in this group discovered the fact that the Acidithiobacillus ferrooxidans could synthesize intracellular electron-dense magnetic particles and this study researched the effect of formation of magnetosomes in bioleaching bacteria under different minerals.
Acidithiobacillus ferrooxidans strainYN-3 was used to reach the effect of physiological characteristics and the change of magnetotaxis under different minerals including pyrite, chalcopyrite, pyrrhotite and sphalerite. The results showed that the physiological characteristics of bacteria had become great difference because of the effect of different minerals.The growth rate of YN-3 was fastest when sphalerite was the source of energy, the next was pyrrhotite and chalcopyrite, YN-3 had the lowest growth rate under the effect of pyrite. Semi-solid flat-panel magnetophoresis indicated that the bacteria were able to orient and migrate to the magnet in artificial magnetic field when pyrite and pyrrhotite as the energy, but there was no such phenomenon when sphalerite and chalcopyrite were energy.
Analyze the response of mpsA, magA, thy and mamB genes of YN-3 under pyrite, chalcopyrite, pyrrhotite and sphalerite by using Real-time PCR. The result of Realtime-PCR showed that these four genes had higher expression under the effect of pyrrhotite and pyrite. On the other hand, because of pyrite is hard to oxidize, the bacteria is difficult to use it as energy. As a result, the growth of YN-3 was slow, it was hard to cultivate on a large scale. According to the factors of physiological characteristics and magnetotaxis of bacteria, we can calculated that pyrrhotite was the best energy mineral for the formation of magnetosomes in YN-3.
In this paper, different concentration of pyrrhotite was used to find the effect of concentration of pyrrhotite to YN-3 on the growth and the formation of magnetosomes. In the experiment, pyrrhotite concentration was 1%,2%,3%,4%,5%. The growth curve showed that YN-3 grew fastest when pyrrhotite concentration was 3%, it had the lowest growth rate with the concentration was 5%. Real-time PCR results showed that the expression of mpsA, magA, and mamB gene tended to increase with the increase of concentration of pyrrhotite. When concentration was 1% to 3%, the expression was increased significantly. While,4% to 5%, the increase tended to be gently. The expression of thy gene had no obvious change with the increase of concentration of pyrrhotite. The growth curve and the result of Real-time PCR showed that 3% was the optimum concentration of pyrrhotite for YN-3.
本课题组发现在氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)等浸矿菌中含有生物控制矿化作用所产生的磁小体。本文主要研究不同矿物作用对浸矿细菌中磁小体形成的影响。
本文以氧化亚铁硫杆菌菌株YN-3为材料,研究在黄铁矿、黄铜矿、磁黄铁矿和闪锌矿等矿物作用下,对YN-3的生长特性及趋磁性产生的影响。结果表明,在不同矿物的作用下,细菌的生长特性有很大不同。在以闪锌矿为能源时,菌体生长速率最快,在第11天,菌量达到最大值2.73X107。其次是磁黄铁矿和黄铜矿,而黄铁矿作用时菌体生长速率最慢。半固体滋泳表明,以黄铁矿和磁黄铁矿作为能源培养的细菌会表现出趋磁性,而利用黄铜矿与铁闪锌矿时无此现象。
利用实时定量PCR技术研究YN-3中与磁小体形成相关的mpsA、magA、thy和mamB基因分别在黄铁矿、黄铜矿、磁黄铁矿和闪锌矿的作用下的表达差异。实时定量PCR结果表明,在以磁黄铁矿和黄铁矿为能源时,四个基因表达量均高于其它两种矿。但是以黄铁矿为能源时,由于矿体难以氧化,菌体生长极为缓慢,难以大量培养收集。综合趋磁性和细菌生长特性两方面因素可知,在磁黄铁矿作用下,YN-3形成磁小体的效果最好,磁黄铁矿为最佳矿物能源。
本文利用不同浓度的磁黄铁矿作用于YN-3,研究不同浓度磁黄铁矿对YN-3的生长状况及对其形成磁小体的影响。实验中磁黄铁矿浓度分别1%、2%、3%、4%、5%。生长曲线结果表明,磁黄铁矿浓度为3%时,菌体生长速度最快。浓度为5%时,菌体生长极为缓慢。实时定量PCR结果表明mpsA, magA和mamB基因随着磁黄铁矿浓度的提高,表达量呈增大趋势。磁黄铁矿浓度为1%-3%时,表达量增加明显。超过3%时,增长趋于平缓。Thy基因在各浓度磁黄铁矿作用下,表达量均较低,没有明显变化。综合生长曲线和实时定量PCR结果两个因素表明,3%为磁黄铁矿对YN-3作用的最适浓度。
Magnetotactic bacteria(MTB) is the name of all kinds of prokaryotes with magnetotaxis, which containing Nano-particles--the magnetosomes. Magnetosomes is an ideal kind of bio-nano-magnetic material, because of its specific properties such as superior crystalline, well biocompatible, uniform nano-sized, non-agglomerated and specific magnetic characteristics.
The researchers in this group discovered the fact that the Acidithiobacillus ferrooxidans could synthesize intracellular electron-dense magnetic particles and this study researched the effect of formation of magnetosomes in bioleaching bacteria under different minerals.
Acidithiobacillus ferrooxidans strainYN-3 was used to reach the effect of physiological characteristics and the change of magnetotaxis under different minerals including pyrite, chalcopyrite, pyrrhotite and sphalerite. The results showed that the physiological characteristics of bacteria had become great difference because of the effect of different minerals.The growth rate of YN-3 was fastest when sphalerite was the source of energy, the next was pyrrhotite and chalcopyrite, YN-3 had the lowest growth rate under the effect of pyrite. Semi-solid flat-panel magnetophoresis indicated that the bacteria were able to orient and migrate to the magnet in artificial magnetic field when pyrite and pyrrhotite as the energy, but there was no such phenomenon when sphalerite and chalcopyrite were energy.
Analyze the response of mpsA, magA, thy and mamB genes of YN-3 under pyrite, chalcopyrite, pyrrhotite and sphalerite by using Real-time PCR. The result of Realtime-PCR showed that these four genes had higher expression under the effect of pyrrhotite and pyrite. On the other hand, because of pyrite is hard to oxidize, the bacteria is difficult to use it as energy. As a result, the growth of YN-3 was slow, it was hard to cultivate on a large scale. According to the factors of physiological characteristics and magnetotaxis of bacteria, we can calculated that pyrrhotite was the best energy mineral for the formation of magnetosomes in YN-3.
In this paper, different concentration of pyrrhotite was used to find the effect of concentration of pyrrhotite to YN-3 on the growth and the formation of magnetosomes. In the experiment, pyrrhotite concentration was 1%,2%,3%,4%,5%. The growth curve showed that YN-3 grew fastest when pyrrhotite concentration was 3%, it had the lowest growth rate with the concentration was 5%. Real-time PCR results showed that the expression of mpsA, magA, and mamB gene tended to increase with the increase of concentration of pyrrhotite. When concentration was 1% to 3%, the expression was increased significantly. While,4% to 5%, the increase tended to be gently. The expression of thy gene had no obvious change with the increase of concentration of pyrrhotite. The growth curve and the result of Real-time PCR showed that 3% was the optimum concentration of pyrrhotite for YN-3.
引文
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