In order to thoroughly investigate the diversity of glacier microorganisms, four DNA extraction methods with different lysis patterns were tested and two screened methods (the Bosshard-Bano method and the Zhou method) were optimized for the most effective form of the filter membrane (cut vs. uncut), the DNA extraction method, and the precipitation method. The two optimized methods were then compared with the commercial Mo-Bio DNA extraction kit, and the results showed that the kit was generally suitable for extraction of microorganism DNA from glacier surface snow. Procedurally, it was found that a modified Bosshard-Bano method (i.e., cutting the filter membrane into pieces, using a specific lysis pattern [lysozyme (5 mg/mL)-protease K (1 mg/mL)-CTAB (1%)-SDS (1%)], performing the extraction only once by chloroform-isoamyl alcohol (24:1), and conducting DNA precipitation by pure ethanol) was also an effective and less expensive method for extraction of microorganism DNA from glacier surface snow.
PeiYing YanShuGui HouTuo ChenShuHong ZhangWeiJun Sun
Over the past 15 years there has been much effort invested in microorganisms of glacial snow and ice on the Tibetan Plateau.These studies include:phenotypic characteristics of recovered isolates;factors(dust,temperature,altitude)influencing microbial abundance,diversity and community in one glacier;distribution of bacterial number,diversity,community along ice core depth;similarities and differences of regionally distributed ice core isolates;seasonal variation of bacterial abundance and diversity.The following need further study.Better methods for more information about the diversity,survival mechanism of glacial microorganism;more research about archaea and fungi;microbial resources;relation of glacial microorganisms with biogeochemical cycle and mass balance;research on altitude distribution.
