PhysOrg
 
   
In subglacial lake, surprising life goes on: Team identifies  species in 
most inhospitable realm
Jul 05, 2013 

 
Lake Vostok, buried under a glacier in Antarctica, is so dark, deep and 
cold  that scientists had considered it a possible model for other planets, a 
place  where nothing could live. 
 
However, work by Dr. Scott Rogers, a Bowling Green State University 
professor  of biological sciences, and his colleagues has revealed a surprising 
variety of  life forms living and reproducing in this most extreme of 
environments. A paper  published June 26 in PLOS ONE details the thousands of 
species 
they  identified through DNA and RNA sequencing. 
"The bounds on what is habitable and what is not are changing," Rogers  
said. 
This is the fourth article the group has published about its Lake Vostok  
investigations. The team included Dr. Paul Morris, biology, who with Scott 
and  doctoral student Yury Shtarkman conducted most of the _genetic analyses_ 
(http://phys.org/tags/genetic+analyses/) ;  former doctoral students Zeynep 
Koçer, now with the Department of Infectious  Diseases, Division of 
Virology, at St. Jude's Research Hospital in Memphis,  performed most of the 
laboratory work; Ram Veerapaneni, now at BGSU Firelands,  Tom D'Elia, now at 
Indian 
River State College in Florida, and undergraduate  student Robyn Edgar, 
computer science. 
Their work was supported by several grants, including two from the National 
 Science Foundation, one from U.S. Department of Agriculture and one from 
the  BGSU Faculty Research Committee. Together, the amount dedicated to the 
project  was more than $250,000. 
When thinking about Lake Vostok, you have to think big. The fourth-deepest 
_lake_ (http://phys.org/tags/lake/)  on Earth, it is  also the largest of 
the 400-some subglacial lakes known in Antarctica. The ice  that has covered 
it for the past 15 million years is now more than two miles  deep, creating 
tremendous pressure in the lake. Few nutrients are available. The  lake lies 
far below sea level in a depression that formed 60 million years ago  when 
the _continental plates_ (http://phys.org/tags/continental+plates/)  shifted 
and cracked. The weather there is so  harsh and unpredictable that 
scientists visiting must have special gear and take  survival training. 
Not only had most scientists believed Lake Vostok completely inhospitable 
to  life, some thought it might even be sterile. 
 
Far from it, Rogers found. Working with core sections removed from the deep 
 layer of ice that accreted from lake water that froze onto the bottom of 
the  glacier where it meets the lake, Rogers examined ice as clear as 
diamonds that  formed in the great pressure and relatively warm temperatures 
found 
at that  depth. The team sampled cores from two areas of the lake, the 
southern main  basin and near an embayment on the southwestern end of the lake. 
"We found much more complexity than anyone thought," Rogers said. "It 
really  shows the tenacity of life, and how organisms can survive in places 
where 
a  couple dozen years ago we thought nothing could survive." 
By sequencing the DNA and RNA from the accretion ice samples, the team  
identified thousands of bacteria, including some that are commonly found in the 
 digestive systems of fish, crustaceans and annelid worms, in addition to 
fungi  and two species of archaea, or single-celled organisms that tend to 
live in  extreme environments. Other species they identified are associated 
with habitats  of lake or ocean sediments. Psychrophiles, or organisms that 
live in extreme  cold, were found, along with heat-loving thermophiles, which 
suggests the  presence of hydrothermal vents deep in the lake. Rogers said 
the presence of  marine and freshwater species supports the hypothesis that 
the lake once was  connected to the ocean, and that the freshwater was 
deposited in the lake by the  overriding glacier. 
The largest number of species overall was found in the area near the  
embayment, including many that are common to freshwater environments, as well 
as  
marine species, psychrophiles and thermophiles. Numerous others were found 
that  remain unidentified. The embayment appears to contain much of the 
biological  activity in the lake. 
"Many of the species we sequenced are what we would expect to find in a  
lake," Rogers said. "Most of the organisms appear to be aquatic (freshwater),  
and many are species that usually live in ocean or lake sediments." 
For Shtarkman, who came to BGSU from St. Petersburg, Russia, the project 
has  proven so engrossing that he foresees a possible lifetime of study around 
it.  "It's a very challenging project and the more you study, the more you 
want to  know," he said. "Every day you are discovering something new and 
that leads to  more questions to be answered. In studying the environmental 
DNA and RNA, we ask  how similar are these sequences to those of sequences 
from organisms already  identified in national databases. We are tracing the 
evolution and the ecology  of the lake itself. 
Before 35 million years ago, Antarctica had a temperate climate and was  
inhabited by a diverse assemblage of plants and animals. About 34 million 
years  ago, Rogers said, a "huge drop in temperature occurred" and ice covered 
the  lake, when it was probably still connected to the Southern Ocean. This 
lowered  sea level by about 300 feet, which could have cut off Lake Vostok 
from the  ocean. The ice cover was intermittent until a second big plunge in 
temperature  took place 14 million years ago, and sea level dropped even 
farther. 
As the ice crept across the lake, it plunged the lake into total darkness 
and  isolated it from the atmosphere, and led to increasing pressure in the 
lake from  the weight of the glacier. While many species probably disappeared 
from the  lake, many seem to have survived, as indicated by Rogers' 
results. 
Rogers's group had worked for several years on identifying and studying  
organisms in the Vostok accretion ice using a procedure involving culturing  
colonies of bacteria and fungi, but the process was slow, especially for  
graduate students who needed results for their theses. 
"We started thinking of doing it a different way," Rogers said. 
Instead of culturing living organisms, they concentrated on sequencing DNA  
and RNA in the ice. These methods, called metagenomics and 
metatranscriptomics,  produced thousands of sequences at a time that were then 
analyzed 
using  computers—procedures referred to collectively as "Big Data" methods. In  
contrast, it usually took years to generate enough cultured organisms for a 
few  dozen sequences. 
The problem changed from having too few sequences to having too many  
sequences to analyze, Rogers said. After two years of computer analysis, the  
final results showed that Lake Vostok contains a diverse set of microbes, as  
well as some multicellular organisms. 
Long before he began using metagenomics and metatranscriptomics to study 
the  ice, Rogers and his team had developed a method to ensure purity. 
Sections of  core ice were immersed in a sodium hypochlorite (bleach) solution, 
then rinsed  three times with sterile water, removing an outer layer. Under 
strict sterile  conditions, the remaining core ice was then melted, filtered 
and refrozen. 
"Using this method, we can assure its reliability almost to 100 percent,"  
Rogers said. 
Eventually, the process rendered pellets of nucleic acids containing both 
DNA  and RNA, able to be sequenced. 
Rogers said the team erred strongly on the conservative side in reporting 
its  results, including only those sequences of which it could be absolutely 
certain  were from the accretion ice, but there are a multitude of others he 
feels are  probably from the lake, opening the door to additional 
investigation. 
The DNA sequences they produced have been deposited in the National Center  
for Biotechnology GenBank database, and will be available to other 
researchers  for further study.

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