LIFE AT THE LIMITS - EXTREME ENVIRONMENTS "HOW CAN THEY SURVIVE?"

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1 COMPARATIVE MICROBIAL GENOMICS ANALYSIS LIFE AT THE LIMITS - EXTREME ENVIRONMENTS "HOW CAN THEY SURVIVE?" Group 5 Kanokwan Inban Lakkhana Khanhayuwa Parwin Tantayapirak

2 Introduction 2

3 Source: 3

4 The Archaea are a group of single-celled microorganisms. They have no cell nucleus or any other organelles within their cells. 4

5 Initially, archaea were seen as extremophiles that lived in harsh environments, such as hot springs and salt lakes, but they have since been found in a broad range of habitats, such as soils, oceans, and marshlands. The archaea exploit a much greater variety of sources of energy than eukaryotes: ranging from familiar organic compounds such as sugars, to using ammonia, metal ions or even hydrogen gas as nutrients. 5

6 Sulfolobus solfataricus 98/2 Sulfolobus islandicus Y.G Metallosphaera sedula DSM 5348 Staphylothermus marinus F1 Sulfolobus islandicus L.S.2.15 Nitrosopumilus maritimus SCM1 Sulfolobus solfataricus P2 Pyrobaculum arsenaticum DSM Thermofilum pendens Hrk 5 Pyrobaculum islandicum DSM 4184 Sulfolobus islandicus L.D.8.5 Sulfolobus islandicus M.16.4 Pyrobaculum aerophilum str. IM2 Hyperthermus butylicus DSM 5456 Crenarchaeota 6

7 Thermoproteus neutrophilus V24Sta Aeropyrum pernix K1 Sulfolobus islandicus M Sulfolobus islandicus M Pyrobaculum calidifontis JCM Sulfolobus tokodaii str. 7 Ignicoccus hospitalis KIN4/I Sulfolobus islandicus Y.N Sulfolobus acidocaldarius DSM 639 Desulfurococcus kamchatkensis 1221n Aciduliprofundum boonei T469 Natronomonas pharaonis DSM 2160 Methanosaeta thermophila PT Halorubrum lacusprofundi ATCC Crenarchaeota Euryarchaeota 7

8 Sulfolobus Archaea Crenarchaeota Thermoprotei Sulfolobales Sulfolobaceae Yellowstone Na9onal Park Ecology: terrestrial volcanic hot springs with op1mum growth occurring at ph temperature of o C + sulfur present acidic environment Genome Structure: circular chromosome that consists of 2,992,245 bp (S. ilandicus) Descrip3on and Significance: - grow either lithoautotrophically or chemoheterotrophically by oxidizing sulfur. - TCA cycle system similar to mitochondria of eukaryotes. Mount St. Helens ** oxidize hydrogen sulfide to sulfate intracellularly used to treat industrial waste water 8

9 Desulfurococcus Archaea Crenarchaeota Thermoprotei Desulfurococcales Desulfurococcaceae Ecology: - deep- sea thermal vents and subterranean hot springs - op1mal temperature for growth is 85ºC. - anaerobic archaea Descrip3on and Significance: - spherical, microns in diameter. - cells are surrounded by an interes1ng protein subunit envelope. - have an interes1ng lasce- like protein structure cell covering - one long flagella 3D image of lasced protein on Desulfurococcus' cell envelope 9

10 Aeropyrum Archaea Crenarchaeota Thermoprotei Desulfurococcales Desulfurococcaceae hyperthermophile ** All species isolated from these environments had been strictly anaerobic, that is, un1l the discovery of Aeropyrum!!! Ecology: solfotaric vent at Kodakara- jima Island in Kyusyu, Japan. - temperatures ranging between 90 to 95 o C, ph 7.0, and a salinity of 3.5% - aerobic archaeon Genome Structure: circular chromosome Descrip3on and Significance: - spherical in shape and are 0.8 to 1.2 microns in diameter - pathways that allow A. to be aerobic gene in the TCA cycle (Krebs' cycle) coding for alpha- ketoglutarate dehydrogenase was not present 10

11 Hyperthermus Archaea Crenarchaeota Thermoprotei Desulfurococcales Pyrodic9aceae hyperthermophile Ecology: - sulfur reducing archaeon that grows between 95 and 106 C and at ph of 7.0. forms H 2 S - sea floor of a hot, solfataric habitat on the coast of São Miguel Island in Azores, Portugal Descrip3on and Significance: - one of the most thermophilic archaea isolated so far (106 C) 11

12 Thermoproteus Archaea Crenarchaeota Thermoprotei Thermoproteales Thermofilaceae hyperthermophile Ecology: acidic hot springs and water holes op1mal growth temperature is 85C. Anaerobes; autotrophic sulfur reduc1on. Genome Structure: Total genome length is nt, and the DNA is double- stranded and circular Descrip3on and Significance: - rod- shaped and reproduce by developing branches on the end of the cell which grow into individual cells. - mo1le by flagella Acidic hot spring, typical environment of Thermoproteus 12

13 Natronomonas Archaea Euryarchaeota Halobacteria Halobacteriales Halobacteriaceae Ecology: aerobic, extremely haloalkaliphilic archaeon. - grows op1mally in 3.5M NaCl and at ph 8.5, - sensi1ve to high magnesium concentra1ons Genome Structure: genome of Natronomonas pharaonis consists of three circular replicons. - chromosome which is 2,595,221 bp in length, - a typical haloarchaeal 131- kb plasmid, and - a unique mul1copy 23- kb plasmid Its choromosome has a high G + C content (63.4%) ** a high propor1on of acidic amino acids (average 19.3%) is found in the proteins of N. pharaonis which results in low isoelectric points (average pi 4.6) adap1ve features of haloarchaea to survive in their hypersaline environment 13

14 Methodology 14

15 1. Downloading Genomes and Prediction of Genes 2. Finding RNA genes and building a 16s rrna tree 3. Genome Atlases and BLAST matrices 4. Pan- Core- genome plots and BLAST atlases 15

16 1. Downloading Genomes and Prediction of Genes 2. Finding RNA genes and building a 16s rrna tree 3. Genome Atlases and BLAST matrices 4. Pan- Core- genome plots and BLAST atlases 16

17 Genome GC content Aciduliprofundum boonei T % Aeropyrum pernix K % Desulfurococcus kamchatkensis 1221n 45.34% %GC Content Halorubrum lacusprofundi ATCC % Hyperthermus butylicus DSM % Ignicoccus hospitalis KIN4/I 56.52% Metallosphaera sedula DSM % Methanosaeta thermophila PT 53.55% Natronomonas pharaonis DSM % Nitrosopumilus maritimus SCM % Pyrobaculum aerophilum str. IM % Pyrobaculum arsenaticum DSM % Pyrobaculum calidifontis JCM % Pyrobaculum islandicum DSM % Staphylothermus marinus F % Sulfolobus acidocaldarius DSM % Sulfolobus islandicus L.D % Sulfolobus islandicus L.S % Sulfolobus islandicus M % Sulfolobus islandicus M % Sulfolobus islandicus M % Sulfolobus islandicus Y.G % Sulfolobus islandicus Y.N % Sulfolobus solfataricus 98/ % Sulfolobus solfataricus P % Sulfolobus tokodaii str % Thermofilum pendens Hrk % Thermoproteus neutrophilus V24Sta 59.91% 17

18 Gene Count Genome Count number of genes genbank refseq prodigal Different Aciduliprofundum_boonei_T469.proteins.fsa Aeropyrum_pernix_K1.proteins.fsa Desulfurococcus_kamchatkensis_1221n.proteins.fsa Halorubrum_lacusprofundi_ATCC_49239.proteins.fsa Hyperthermus_butylicus_DSM_5456.proteins.fsa Ignicoccus_hospitalis_KIN4_I.proteins.fsa Metallosphaera_sedula_DSM_5348.proteins.fsa Methanosaeta_thermophila_PT.proteins.fsa Natronomonas_pharaonis_DSM_2160.proteins.fsa Nitrosopumilus_maritimus_SCM1.proteins.fsa Pyrobaculum_aerophilum_str_IM2.proteins.fsa Pyrobaculum_arsenaticum_DSM_13514.proteins.fsa Pyrobaculum_calidifontis_JCM_11548.proteins.fsa Pyrobaculum_islandicum_DSM_4184.proteins.fsa Staphylothermus_marinus_F1.proteins.fsa Sulfolobus_acidocaldarius_DSM_639.proteins.fsa Sulfolobus_islandicus_LD85.proteins.fsa Sulfolobus_islandicus_LS215.proteins.fsa Sulfolobus_islandicus_M1425.proteins.fsa Sulfolobus_islandicus_M1627.proteins.fsa Sulfolobus_islandicus_M164.proteins.fsa Sulfolobus_islandicus_YG5714.proteins.fsa Sulfolobus_islandicus_YN1551.proteins.fsa Sulfolobus_solfataricus_98_2.proteins.fsa Sulfolobus_solfataricus_P2.proteins.fsa Sulfolobus_tokodaii_str_7.proteins.fsa Thermofilum_pendens_Hrk_5.proteins.fsa Thermoproteus_neutrophilus_V24Sta.proteins.fsa refseq-prodigal 18

19 Gene Length Genome genbank refseq prodigal Mean StdDev Min Max Mean StdDev Min Max Mean StdDev Min Max Aciduliprofundum_boonei_T469.proteins.fsa Aeropyrum_pernix_K1.proteins.fsa Desulfurococcus_kamchatkensis_1221n.proteins.fsa Halorubrum_lacusprofundi_ATCC_49239.proteins.fsa Hyperthermus_butylicus_DSM_5456.proteins.fsa Ignicoccus_hospitalis_KIN4_I.proteins.fsa Metallosphaera_sedula_DSM_5348.proteins.fsa Methanosaeta_thermophila_PT.proteins.fsa Natronomonas_pharaonis_DSM_2160.proteins.fsa Nitrosopumilus_maritimus_SCM1.proteins.fsa Pyrobaculum_aerophilum_str_IM2.proteins.fsa Pyrobaculum_arsenaticum_DSM_13514.proteins.fsa Pyrobaculum_calidifontis_JCM_11548.proteins.fsa Pyrobaculum_islandicum_DSM_4184.proteins.fsa Staphylothermus_marinus_F1.proteins.fsa Sulfolobus_acidocaldarius_DSM_639.proteins.fsa Sulfolobus_islandicus_LD85.proteins.fsa Sulfolobus_islandicus_LS215.proteins.fsa Sulfolobus_islandicus_M1425.proteins.fsa Sulfolobus_islandicus_M1627.proteins.fsa Sulfolobus_islandicus_M164.proteins.fsa Sulfolobus_islandicus_YG5714.proteins.fsa Sulfolobus_islandicus_YN1551.proteins.fsa Sulfolobus_solfataricus_98_2.proteins.fsa Sulfolobus_solfataricus_P2.proteins.fsa Sulfolobus_tokodaii_str_7.proteins.fsa Thermofilum_pendens_Hrk_5.proteins.fsa Thermoproteus_neutrophilus_V24Sta.proteins.fsa

20 1. Downloading Genomes and Prediction of Genes 2. Finding RNA genes and building a 16s rrna tree 3. Genome Atlases and BLAST matrices 4. Pan- Core- genome plots and BLAST atlases 20

21 Sulfolobus sp. 16s rrna tree Pyrobaculum sp. 21

22 1. Downloading Genomes and Prediction of Genes 2. Finding RNA genes and building a 16s rrna tree 3. Genome Atlases and BLAST matrices 4. Pan- Core- genome plots and BLAST atlases 22

23 Thermoproteus neutrophilus V24Sta Sulfolobus sp. Pyrobaculum sp. BLAST Matrices 23

24 1. Downloading Genomes and Prediction of Genes 2. Finding RNA genes and building a 16s rrna tree 3. Genome Atlases and BLAST matrices 4. Pan- Core- genome plots and BLAST atlases 24

25 Pan- Core genome plots 25

26 Pan- Core genome plots 26

27 THANK YOU 27

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