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Exploring the Sulfur Nutrient Cycle Using the Winogradsky Column

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Exploring the Sulfur Nutrient Cycle Using the Winogradsky Column -4 II 0- H 0 0 Downloaded from http://online.ucpress.edu/abt/article-pdf/67/6/348/52459/4451860.pdf by guest on 23 September 2021 Exploring the Sulfur Nutrient Cycle Using the Winogradsky Column BRIANROGAN MICHAELLEMKE MICHAELLEVANDOWSKY THOMAS GORRELL ETo fullyunderstand the workingsof thebiolog- ogy to help meet the many demands and standards ical world, it is importantthat students have a fun- that are part of today'sscience classes. damentalsense of the naturalcycles that providethe The Winogradsky column is a glass or clear nutrients and energy that power life, as well as a plastic column, filled with enriched soil or sedi- sense of how these systems evolved.Many teachers ment. When developed, it has an anaerobiclower cover carbon cycles and emphasize microbial zone and aerobicupper zone that allow growth of processes when reviewingthe complexitiesof nitro- microorganismsunder conditions similar to those gen cycling,but often the sulfur cycle, if covered,is found in sediments and water rich in nutrients done so Theremay be many reasonsfor this: briefly. (Sylvia et al., 1998). Often teachers simply convey time limitations,the element is less prevalentthan the message that differentmicroorganisms exist in others as a biological constituent, or the topic is differentstrata of the column and that some live in thought to be too complex. However,teaching the the aerobicand some in anaerobiczones. However, sulfur in conjunction with the classic cycle this is reallywhere the discoverybegins ratherthan Winogradskycolumn exercisepresents the opportu- ends! Explainingthe complexitythat lies within the nity to cover severalimportant topics simultaneous- depths of the ecosystem allows deeper insights into ly. The exercise links microbialprocesses, concepts the microbialworld. of biodiversity,inorganic chemistry, biogeochemical cycling,evolution, microbiology, and microbialecol- In the laboratory, the Winogradsky column demonstrates how the metabolic diversity of prokaryotes transforms sulfur, an essential con- stituentof livingmatter and an abundantelement in BRIAN ROGAN teaches at The New Jewish High School, the Earth'scrust (Stanier et al., 1976), to different Waltham, MA 02453. MICHAELLEMKE is in the Biology Departmnent,University of Illinois at Springfield,Springfield, forms with varying redox states, thus supplying IL 62703-5407. MICHAELLEVANDOWSKY and THOMASGORRELL nutrientsand/or energyto the organism.The micro- work at Haskins Laboratory,Pace University,New York,NY bial assemblagethat develops in the column spatial- 10038-1598. ly separates organisms into distinct layers several 348 THEAMERICAN BIOLOGY TEACHER, VOLUME 67, NO.6,AUGUST 2005 centimeters thick even though in the environment 3. discuss how organisms may be isolated and establishment of similar layers of different organisms grown in a homemade anaerobicchamber. would typicallyexist in a few millimetersof sediment. Useful laboratoryand educationalextensions of the The Winogradsky column creates conditions that exerciseare also discussed. expand the volume of natural processes, allowing a clear view of naturally-occurringphenomena. Soil sam- ples are collected fromwetland habitats,amended with Materials & Methods simple inorganicand organicmaterials, then exposed to Materials needed for the construction of the light as an externalenergy source. The results are a mul- Winogradskycolumn are simple and common. They ticoloredcolumn of soil and water,each color linked to include: a chemical or biological process. The defined zones of microbes develop form accordingto concentrationgra- * a clear glass or plastic container(e.g., a smooth- dients of oxygen, sulfur,nutrients, and light. Each func- sided, quartplastic waterbottle at least 15 cm in tional microorganism group is dependent on other height and 5 cm in diameter.Plastic bottles are functionalmicrobial groups for development. flexible and can be manipulated to allow for extractionof species for culturing.Very tall con- The Winogradsky column was developed and tainers requirelonger development periods and Downloaded from http://online.ucpress.edu/abt/article-pdf/67/6/348/52459/4451860.pdf by guest on 23 September 2021 named after Sergei Winogradsky (1856-1953), a the bacteriamay be more difficultto extract.) Russian microbiologist.He studied the complex inter- * actions between environmentalconditions and micro- plastic film and a rubberband bial activitiesusing soil enrichmentto isolate pure bac- * a wooden dowel terial cultures (Madigan et. al, 2000). Louis Pasteur, * a sulfur source (e.g., calcium sulfate,magnesium Robert Koch, and other scientists isolated cultures for sulfate, or egg yolk added at about 1-2%of the study, but Winogradsky'swork was one of the first to soil weight) study microorganismsin mixed enrichment cultures. The fact that the exercise works under a wide range of * an inorganic carbon source (calcium carbonate circumstancesis a testamentto the near ubiquityof cer- [e.g., chalk or limestone] or baking soda may be tain functionalgroups of microbes. added to 1-2%of soil weight) * hydricsoil (e.g., pond mud or shallow riversedi- The Winogradsky column may also be used to ment collected near the surface) demonstrate aspects of the earliest, sulfur-basedlife forms found on Earth. In an article in Nature,Nisbet * cellulose (e.g., shreddedpaper towels) (2000) paints a picture of an environment of early * a 60-75 watt light source organismsin the Archeanperiod 2.5 to 4 billion years * from the same source as the sediment ago that are analogous to those found in hydrothermal water vents. Hydrothermalvents were first observed by the Breakup soil clumps and sieve out largerdebris so submersiblevehicle Alvin that exploredthe Mid-Atlantic the column can be packed evenly.The muddy mixture spreading ridge where the North American and should be stirred to gain a uniform consistency and European plates are inexorably moving apart. This should include the sulfurand inorganiccarbon sources. observationmarked the discoveryof a system that may Place a 2-3 cm layerof the mud mixturein the column, have remainedintact since its formationas an ancient add the source of cellulose, and stir and pack with the biotic system utilizingsimple nutrientcycles as an ener- dowel.Add as much of the mixtureas needed, 2-3 cm at gy source. In a sense, by creatinga Winogradskycol- a time, with gentle tampingwith the dowel to force out umn, we are modeling ancient environments,though trappedair, until the tower of mud is about 5 cm from perhaps at lower temperatures. the top of the container.Your last layer should be 2-3 cm of water. Cover the opening with plastic film and Constructinga Winogradskycolumn from simple Place the column next to a homemade materialsis only one of the exercises dis- secure with a rubberband. low heat, moderate intensity light cussed here. We also present a technique that uses the continuously-lit, sure the column does not overheat. respirationof seed germinationto allow the reciprocal source, making process, anaerobic metabolism, to occur in a simple Examinethe columns weekly for at least a month, glass baking dish. The objectivesof this articleare to: recording changes in color as they occur. For the time 1. describe the microbial ecosystem of the Winogradsky column to be successful, enough Winogradskycolumn as a tool for studying the must be allowed for the cultures to develop. The columns may show growth in a week, as indicated by cycling of sulfur formationof a black color near the bottom and disinte- 2. explainhow use of the columnmay illustrate some gration of the cellulose (paper), but will probablynot featuresof developmentof earlylife on Earth fully form and stabilize for four weeks or more. The SULFURNUTRIENT CYCLE 349 ideal situation would be for students to investigatethe Isolation & Culturing of column over the course of the year studying ecology, microbiology,biodiversity, evolution, and other biologi- Organisms from the cal themes. Winogradsky Column Variations When pigmented patches are visible in the column (Figure 1), one can attemptto isolate some of the organ- Constructionand developmentof the Winogradsky isms. Samplingcan occur at weekly intervals to check column incorporatesseveral variables. With just a few succession or can be done at the end of the project to changes, differentcolumns can be created to compare see the final flora of bacteria that develops. Sampling growth rates, microbial populations, and ecological may be done using a standardbacteriological nichrome diversity. wire loop or hypodermic needle (pierced through the * Sulfur Source side of the plastic container), however distinction between the microbes and mud is often difficult.Look To illustratethe importanceof the type of sulfur for Beggiatoa(Figure 2A) or Thiobacillusin the water- as a substrate,sodium sulfide or elementalsulfur sediment interface.Flagellated organisms from the col- can be added in place of a sulfate. This should umn, such as Rhodopseudomonas(Figure 2B) or those Downloaded from http://online.ucpress.edu/abt/article-pdf/67/6/348/52459/4451860.pdf by guest on 23 September 2021 reduce the growth of the sulfate-reducingbacte- with sulfur inclusion bodies, like Chromatiumand ria and alter the composition of the microbial Thiospirillium(Figure 2D) are more easily identified if community. you carefully adjust the contrast on a standard light * Acidity compound microscope or have a phase-contrastmicro- scope. The green sulfur (Figure 2C) and sulfate-reduc- Acid affectsthe biotic component of our environ-
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