厌氧原生动物的内共生甲烷菌及其氢酶的细胞化学研究

STUDIES ON METHANOGENIC ENDOSYMBIONTS IN ANAEROBIC PROTOZOA, WITH REFERENCE TO THE CYTOCHEMICAL ASPECTS OF HYDROGENASE

  • 摘要: 阿米巴鞭毛虫Psalteriomonas lanterna生活史分为鞭毛虫和阿米巴阶段。其培养液2%(V/V)氧时,细胞失去了内共生甲烷菌,继续培养在1%氧中,则鞭毛消失,小阿米巴出现。在1%氧条件下,阿米巴细胞生长较快,密度约1.2×104cells/ml,氧含量2.5%生长被抑制,超过7.5%细胞则死亡。无氧状态生长缓慢,加入甲酸甲烷杆菌(Methanobacterium formicicum Strain MsL)后,出现细胞密度增加的趋势。外萤光显微镜和电镜观察以及水洗P.lanterna后,培养液中生长的甲烷产物的事实证明;其细胞内含的萤光菌是甲烷杆菌(种类待定)。细胞化学染色法对细胞中氢酶的活性定位证实P.lanterna所含的类似微体的细胞器是氢体。染色结果:低浓度的戊二醛固定细胞,外加H2和苯异噻唑酞肼苯乙烯四唑(BspT)化合物,细胞的氢体膜周围有电子致密点生成。整个过程是在非常严格的厌氧条件下进行的。对照组以N2代替H2的细胞无上述反应。

     

    Abstract: The life cycle of amoebaflagellate Psalteriomonas lanterna includes two stages: flagellate and amoeba. When 2% oxygen was added to the headspace of the culture, the flagellate lost its symbiont metbanogenic bacteria. When flagellates without endosymbionts were cultured with 1% oxygen, small amoeba appeared. When cultured with 1% oxygen, amoeba cells grew quickly, the concentration of cells was about 1.2×10 ceUs/ml, and inhibition occurred at oxygen concentrations over 2.5%. The cells died when the oxygen level exceeded 7.%. Amoebae grew poorly in anaerobic culture. Addition of Methanobacterium formicicum Strain MsL to the amoebae resulted in higher concentrations of cells. The fluorescent endosyrnbionts in the sapropelic amoebaflagellate were proved to be methanogenic bacteria (species not identified). This was indicated by epifluorescence and electron microscopic observations, and the fact that methane was produced in the culture medium when the protozoa were washed in water. Cytochemical staining method revealed the presence of hydrogenase activity in the microbodies of P. lanterna without endosymbionts, suggesting that the microbody-like organelles are hydrogenesomes. After fixation of the cells in low concentrations of glutaraldehyde and incubation in the presence of hydrogen and the tetrazolium compound, 2-(2′-benzothizazolyl)-5-styryl-(-3-4-phthalhydrazidyl)-tetrazolium chloride, an electron-dense deposit was produced in the hydrogenosomes. In the control cells exposed to nitrogen, no deposit was found.

     

/

返回文章
返回