Name: establishment of microbial eukaryotic enrichment cultures from a chemically stratified antarctic lake and assessment of carbon fixation potential
Uploaded: 2012-04-20T13:00:00
Description: Watch this Scientific Journal Video about Establishment of Microbial Eukaryotic Enrichment Cultures from a Chemically Stratified Antarctic Lake and Assessment of Carbon Fixation Potential at JoVE.com

Forfatterne takker J. Priscu, A. Chiuchiolo og McMurdo Filterpressens limnologi team for bistand i innsamling og bevaring av prøvene i Antarktis. Vi takker Ratheon Polar Services og Phi helikoptre for logistisk støtte. Lette mikrografer ble generert i Miami senter for avansert mikroskopi og Imaging Center. Dette arbeidet ble støttet av NSF Office of Polar radioprogrammer Grants 0631659 og 1056396.

1. Eksempel Oppkjøp <ol><li> Velg og forberede målepunktet en dag før prøvetaking vannsøylen. Dette vil tillate stratifiserte lagene i vannsøylen for å reformere etter forstyrrelse på grunn av boring og is-hull smeltende. Identifiser plassering av borestedet ved GPS. </li><li> For å få tilgang vannsøylen, begynne med å bore et hull gjennom isen med en Jiffy is Auger festet til en 4-tommers Jiffy flytur forlengelse og skjæring bit. For å hindre frysing av boret i hullet, prøv å unngå boring i flytende ...

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Nyere molekylære studier har rapportert høy diversitet av encellede eukaryote tvers av en rekke tre miljøer, 19, 20, men på grunn av mangel på isolater over hele spekteret av protist habitater de funksjonelle roller disse individuelle arter i næringskjeder i stor grad ukjent. I denne studien har vi beskrevet metoder for å berike for mikrobielle eukaryote arter stiller metabolsk allsidighet fra en relativt undersampled miljø, en permanent islagte Antarktis innsjø. Kulturer beriket fra ulike prøvetaki...

<table border="1"><tbody><tr><td> Navn på reagensen </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> BBM </td><td> Sigma </td><td> B5282 </td><td></td></tr><tr><td> BG11 </td><td> Sigma </td><td> C3061 </td><td></td></tr><tr><td> F / 2 </td><td> Sigma </td><td> G9903 </td><td></td></tr><tr><td> GF / F filter, 25 mm </td><td> Fisher Scientific </td><td> 09-874-64 </td><td></td></tr><tr><td> GF / F filter, 47 mm </td><td> Fisher Scientific </td><td> 09-874-71 </td><td></td></tr><tr><td> Polyetersulfon filter, 0,45 mikrometer pore, 47 mm </td><td> Pall Life Sciences </td><td> 61854 </td><td></td></tr><tr><td> Steril cellekultur kolbe, 25 cm 2 </td><td> Corning </td><td> 430639 </td><td></td></tr><tr><td> Diurnal vekst kammer </td><td> VWR </td><td> 35960-076 </td><td></td></tr><tr><td> Zirconia / silika perler, 0,1 mm diamter </td><td> BioSpec Produkter </td><td> 11079101z </td><td></td></tr><tr><td> Mini-Bead beater </td><td> BioSpec Produkter </td><td> 3110BX </td><td></td></tr><tr><td> Skrue-cap mikrosentrifuge tube (1,5 mL) </td><td> USA Scientific </td><td> 1415-8700 </td><td></td></tr><tr><td> NaH 14 CO 3 </td><td> ViTrax </td><td> VC 194 </td><td> Hold alikvoter av 400 mL ved -20 ° C </td></tr><tr><td> RuBP </td><td> Sigma </td><td> R0878-100mg </td><td> Løs opp i 10 mM Tris-propionsyre (pH 6,5) </td></tr></tbody></table>

establishment of microbial eukaryotic enrichment cultures from a chemically stratified antarctic lake and assessment of carbon fixation potential

Lake Bonney er en av utallige permanent islagte innsjøer som ligger i McMurdo Dry Valleys i Antarktis. Den flerårige isen opprettholder en kjemisk stratifisert vannsøylen og i motsetning til andre innlandet vannforekomster, hovedsakelig hindrer ekstern tilførsel av karbon og næringsstoffer fra bekker. Biota er utsatt for en rekke miljømessige belastninger, blant annet helårs alvorlig næringsmangel, lave temperaturer, ekstrem skygge, hypersalinity, og 24-timers mørke vinteren en. Disse ekstreme miljøforhold begrense biota i Lake Bonney nesten utelukkende til mikroorganismer to. Encellede mikrobielle eukaryoter (kalt &quot;protister&quot;) er viktige aktører i biogeokjemiske sykler tre og spiller viktige økologiske roller i sykling av karbon i de tørre dalen innsjøer, opptar både grunnskoler og videregående roller i akvatisk næringskjeden. I de tørre dalen akvatiske næringskjeden, protister som løser jegnorganic karbon (autotrophy) er de største produsentene av organisk karbon for organotrophic organismer 4, 2. Phagotrophic eller heterotrofe protister stand til inntak bakterier og mindre protister opptre som toppredatorer i næringskjeden 5. Sist, er en ukjent andel av protist befolkningen i stand til kombinerte mixotrophic metabolisme 6, 7. Mixotrophy i protister innebærer evnen til å kombinere fotosyntetiske evne med phagotrophic inntak av byttedyr mikroorganismer. Denne formen for mixotrophy forskjellig fra mixotrophic metabolisme i bakterielle arter, noe som vanligvis innebærer opptak oppløste karbon molekyler. Det er for tiden svært få protist isolater fra permanent is-capped polare innsjøer, og studier av protist mangfold og økologi i dette ekstreme miljøet har vært åtte begrenset, 4, 9, 10, 5. En bedre forståelse av protist metabolsk allsidighet i det enkle tørre dalen innsjø næringskjeden vil hjelpemiddel i utvikling av modeller for role av protister i den globale karbonsyklusen. Vi ansatt en berikelse kultur tilnærming for å isolere potensielt phototrophic og mixotrophic protister fra Lake Bonney. Prøvetaking dyp i vannsøylen ble valgt basert på plasseringen av primærproduksjonen maxima og protist fylogenetisk fire mangfold, 11, samt variasjon i viktige abiotiske faktorer som påvirker protist trofiske moduser: grunt prøvetaking dyp er begrenset for store næringsstoffer, mens dypere prøvetaking dybder er begrenset av lys tilgjengelighet. I tillegg ble innsjø vannprøver supplert med flere typer vekstmedier for å fremme veksten av en rekke phototrophic organismer. RubisCO katalyserer det hastighetsbegrensende trinnet i Calvin Benson Bassham (CBB) syklus, den viktigste veien der autotrophic organismer fikse uorganisk karbon og gi organisk karbon for høyere trofiske nivåer i akvatiske og terrestriske næringskjeder 12. I denne studien, we brukte et radioisotop analysen modifisert for filtrerte prøver 13 til overvåke maksimalt carboxylase aktivitet som en proxy for karbon fiksering potensial og metabolsk allsidighet i Lake Bonney berikelse kulturer.

Watch this Scientific Journal Video about Establishment of Microbial Eukaryotic Enrichment Cultures from a Chemically Stratified Antarctic Lake and Assessment of Carbon Fixation Potential at JoVE.com

Establishment of Microbial Eukaryotic Enrichment Cultures from a Chemically Stratified Antarctic Lake and Assessment of Carbon Fixation Potential

Mikrobielle eukaryoter er både en kilde til photosynthetically-derived carbon og topp rov arter i permanent islagte Antarktis innsjøer. Denne rapporten beskriver en berikelse kultur tilnærming for å isolere metabolsk allsidige mikrobielle eukaryoter fra Antarktis innsjø, Lake Bonney, og vurderer uorganisk karbon fiksering potensial ved hjelp av en radioisotop analysen for ribulose-bifosfatase-karboksylase/oksygenase-proteinet-1 ,5-bisphophate carboxylase oxygenase (RubisCO) aktivitet.

Etablering av Mikrobielle Eukaryote berikelse kulturer fra en kjemisk Stratifisert Antarktis Lake og vurdering av Carbon Fiksering Potential

Lake Bonney is one of numerous permanently ice-covered lakes located in the McMurdo Dry Valleys, Antarctica. The perennial ice cover maintains a ...

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