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Prochlorococcus : probes; distribution; Chile dot blots; Indian Ocean cruise Synechococcus cultures : status; multilocus sequencing approach 13 C/ 14 C.

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Presentation on theme: "Prochlorococcus : probes; distribution; Chile dot blots; Indian Ocean cruise Synechococcus cultures : status; multilocus sequencing approach 13 C/ 14 C."— Presentation transcript:

1 Prochlorococcus : probes; distribution; Chile dot blots; Indian Ocean cruise Synechococcus cultures : status; multilocus sequencing approach 13 C/ 14 C activity expts Erythrosphaera marina, new name Crocosphaera watsonii WH8501 2-5 um size; fixes N 2 (maximally at night) temp. range for growth 26°C -32°C Cyanothece sp strains WH 8902 and 8904; benthic & fix N 2 Prochlorococcus and Synechocoocus cultures

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4 Strain Clonal Colour Motility ntcA RFLP 16S sequence type WH7803 + Red - VII + WH8018 + Red - VIII + WH8103 + Orange + IX + Dim + Red - VII + Bright + Green - N/A + Minos1 (RCC45) - Red - + Minos2 (RCC47) - Orange - + Minos11 (RCC61) - Red - + Minos12(RCC62) - Orange - + Almo3 (RCC43) - Red - + Oli31 (RCC44) - Orange/Red - + Eum14 (RCC37) - Orange - + Max42 (RCC30) - Orange - + RCC30 + Orange - RCC37 - Orange - RCC43 - Red - RCC44 - Orange/Red XII - RCC45 + Red - RCC47 - Orange - RCC61 + Red - RCC307* + Red X + RCC308 + Red X - RCC309 + Red X - RCC310** + Orange IX + RCC311** + Orange IX + RCC312** + Orange IX + RCC313** + Orange IX + Analysis of Synechococcus strains I * identical 16S rRNA sequence to Minos 1 over 550 bp ** identical 16S rRNA sequence to Minos 2 over 550 bp

5 Analysis of Synechococcus strains II Blanes Bay Strain Clonal ntcA RFLP type BL1 -VIII BL5 -VIII BL6 -VIII BL7 -VII/VIII BL8 - XI (similar to VIII) BL10 -VII BL11 -VIII BL29 -VIII BL30 -VIII BL31 -VIII BL32 -VII BL5/4 -VIII BL6/5 -VIII BL7/4 -VII/VIII BL8/4 -VII BL17/4 -VIII BL18/4 -VIII BL19/5 -VIII BL20/4 -VIII

6 Analysis of Synechococcus strains IV Prosope Cruise Strain two rounds of growing clonally plating achieved in liquid medium 77-2 (RCC329)   153-3   53-4   135-6 lost - 53-19  possibly 157-13   117-1  possibly 32-1  x 97-4 lost - 157-21   25-2  x 41-3   37-2 lost - 85-6  possibly

7 Strain depth oftwo rounds of growth in liquid? isolationplating achieved RCC316 (37-2)110 lost - RCC317 (85-6) 90  possibly RCC318 (41-3) 65   RCC319 (157-21) 15   RCC320 (157-13) 15   RCC321 (32-1) 5  x RCC322 (117-1) 5  possibly RCC323 (97-4) 5 lost - RCC324 (53-4) 5   RCC325 (153-3) 25   RCC326 (53-19) 5  possibly RCC327 (25-2) 25  x RCC328 (135-5) 30 lost - RCC329 (77-2) 5   Analysis of Synechococcus strains IV Prosope Cruise

8 RS9902con.seq RS9911con.seq RS9908con.seq BL6pErrev.SEQ BL1pErrev.SEQ RS9901con.seq He24con.seq RS9906con.seq RS9918con.seq Syn8101.SEQ 8103cont.SEQ Min2cor.SEQ Min12cor.SEQ Almo3cor.SEQ CC9311cor.SEQ 80187cor.SEQ SynWH7805.SEQ 78035cont.SEQ Oli31cont.SEQ BL8pErrev.seq ProNATL1.SEQ ProNATL2.SEQ ProPCC9511.SEQ ProMED4.SEQ BL3pErrev.SEQ BL7pErrev.SEQ ProMIT9202.SEQ ProMIT9215.SEQ ProTATL2.SEQ DYF33con.seq MIO31con.seq ProPAC1.SEQ MIO43con.seq DYF13con.seq ProMIT9211.SEQ MIO60con.seq ProMIT9313.SEQ Min11cont.SEQ BL5pErrev.SEQ BL2pErrev.SEQ Min1cor.SEQ Brightcont.SEQ CA Syns Pros

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11 ntcA sequence variation Between Syns and Pros 22-33 % variation Within Pros: 5-10% variation within clusters; 17-26% variation between clusters Within Syns: 2-3% variation within clusters; 20-23% variation between clusters

12 Workpackage 2 : Clones libraries Solve problem of cyanobacteria/ plastid clone libraries - potential use of a 23S rRNA primer Should we construct other plastid/cyano clone libraries? Where? Low oxygen zone - Chile Arabian Gulf Screening of samples prior to clone libraries by DGGE Should we do more eukaryotic clone libraries? Where ? Bloom evolution (Roscoff) Oslo fjord in conjunction with TEM PROSOPE cruise after screening of samples Red Sea from 2000 workshop Antarctic Should we probe clone libraries from underrepresented groups (cf Haptophytes etc...) Which clones should we completely sequence?

13 Workpackages 3-4 : Probes List of groups to be targeted Suggestion: for FISH it would be best to only have Class level probes while for DNA chips we could have both class level and species specific probes knowing that we have no limitation in terms of the number of probes that can be used. See below for rough list based on what we found in clone libs. List of probes to be designed Classes Genus, species What will be the focus of each partner in terms of probe validation? Probe detection: FISH by microscopy and flow cytometry Quantitative PCR Dot blot DNA chips

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