1. Pigments in Prasinophyceae

2. BLA148/9 BLA78/7 RCC287 BLA82/7 RCC343 (Ostreococcus IV) RCC116 (O. tauri) RCC344 (Ostreococcus IV) RCC371 (Ostreococcus IV) RCC356 (Ostreococcus IV) BLA122 RCC393 (Ostreococcus II) RCC136 (Prasinococcus) RCC137 (Prasinoderma) Chlorodendrales Mamiellales Prasinococcales

3. Final Summary The two types of pigmentation in Prasinophyceae according to Egeland et al (1995) were clearly identified by cluster analysis In addition, the clones could be grouped into the three established orders. However, the pigment data also point to a large discrepancy within the order Chlorodendrales (new order?) There were important non-common features within each of these two groups. Perhaps the most remarkable were the following: Presence of Chl c3 in two clones (BLA122, RCC393) Simultaneous presence of Prasinoxanthin and Lutein in two clones (RCC136, RCC137) Presence of MgDVP in a Chlorophyceae-type prasinophyceae (BLA78/7) Absence of Violaxanthin in one clone (RCC137)

4. Final Summary The two types of pigmentation in Prasinophyceae according to Egeland et al (1995) were clearly identified by the cluster analysis In addition, the clones could be grouped into the three established orders. However, the pigment data also point to a large discrepancy within the order Chlorodendrales (new order?) There were important non-common features within each of these two groups. Perhaps the most remarkable were the following: Presence of Chl c3 in two clones (BLA122, RCC393) Simultaneous presence of Prasinoxanthin and Lutein in two clones (RCC136, RCC137) Presence of MgDVP in a Chlorophyceae-type prasinophyceae (BLA78/7) Absence of Violaxanthin in one clone (RCC137) It's the rule

5. 13 pigments and 13 strains

6. ChlcMgDVPUriolideViollikeNeoxPrasViolMmonalUnid-z1Unid-z2AntheraxZeaxLutDihydroChlbChla  -car-likeUnid M1  -car  -car Tetraselmis sp. RCC5000.0180.0310.0240.0080.0140.1380.6551.0000.0620.0000.191 RCC287 (CCMP1205 = RCC15) NOUM150.0180.0740.1310.0510.0420.3821.3131.0000.0260.053 Pyramimonas parkeae CCMP7240.0010.0210.0770.0020.0020.0700.6771.0000.0110.088 Tetraselmis sp. RCC2340.0220.0390.0330.0570.0060.0050.0960.8121.0000.0090.0200.062 Crustomastix stigmatica0.0230.0080.0560.0550.0170.0650.0380.7861.0000.0340.271 Nephroselmis sp RCC4990.0070.0230.0300.0530.0360.0330.0180.7231.0000.0200.0410.075 Nephroselmis pyriformis CCMP7170.0150.0440.0210.0770.0160.0190.0220.6981.0000.0050.0400.025 RCC5010.0620.0730.0220.0390.1360.0470.0380.0100.0230.0190.0200.0050.0320.6271.0000.0270.0000.044 Ostreococcus tauri RCC1160.0610.0790.0160.0450.1910.0440.0390.0140.0250.0510.0500.0030.0320.6331.0000.0280.0000.098 Ostreococcus sp. RCC3560.0670.0860.0190.0450.1770.1020.0420.0040.0100.0350.0750.0070.0390.6371.0000.0310.0030.042 Ostreococcus sp. RCC3440.0570.0680.0150.0410.1540.1010.0400.0150.0220.0310.0250.0020.0270.6181.0000.0330.039 Ostreococcus sp. RCC3710.0600.0740.0150.0440.1560.1050.0430.0180.0210.0160.0280.0010.0310.6251.0000.0370.039 Ostreococcus sp. RCC3430.0630.0770.0160.0430.1860.0550.0430.0230.0270.0470.0330.0010.0280.6481.0000.0300.036 Bathycoccus prasinos RCC1130.0410.0470.0180.0180.0970.0780.0300.0070.0230.0020.0170.0030.0180.4621.0000.0130.029 Ostreococcus sp. RCC3930.0200.0240.0800.0100.0600.2120.1230.0330.0200.0220.0550.0380.0060.0450.6721.0000.0350.046 Micromonas sp.RCC4180.0800.0850.0160.0360.1300.0890.0380.0110.0200.0200.0600.0100.0070.8121.0000.0280.032 Micromonas pusilla RCC372 (clade CCMP 489)0.0150.0450.0840.0250.0440.1680.1390.0570.0110.0290.0130.0160.0060.0180.8101.0000.0260.031 Micromonas pusilla RCC114 (clade CCMP490)0.0720.0960.0260.0450.1910.1030.0530.0120.0320.0190.0130.0060.0100.7681.0000.0310.0020.056 Micromonas RCC498 (clade CCMP490)0.0670.0860.0330.0370.1600.1490.0470.0150.0360.0140.0120.0030.0050.7971.0000.0220.0030.035 Mamiella sp. RCC3910.0600.0720.0210.0430.1890.0740.0100.0150.0240.0110.0190.0080.0520.8961.0000.0340.0270.055 Mantoniella squamata RCC4170.0680.0750.0290.0370.1500.0680.0450.0130.0330.0060.0100.0030.0210.6441.0000.0260.0200.101 Mantoniella sp. RCC3950.0650.0730.0280.0310.1230.0560.0430.0080.0280.0030.0060.0040.0090.6221.0000.0210.0130.049 Ostreococcus sp. RCC1410.0200.0560.0760.0430.0370.1440.0190.0590.0090.0330.0040.0251.0321.0000.0270.0140.016 Ostreococcus oceanicus RCC1430.0290.0360.0720.0360.0380.1460.0280.0590.0090.0300.0040.0301.0221.0000.0320.0120.018 Prasinococcus cf capsulatus RCC136/CCMP14070.0200.0250.0810.0150.0600.1920.0470.0080.0160.0160.1800.0530.7131.0000.0240.030 Prasinococcus capsulatus CCMP11920.0980.0970.0170.0760.1830.0350.0200.0230.0010.0080.0150.0050.6201.0000.0060.019 Prasinococcus sp RCC3420.0980.1020.0210.0680.2090.0350.0080.0190.0080.0140.1110.9531.0000.0320.011 Prasinoderma RCC137/CCMP12200.1090.0640.0150.0690.1950.0190.0260.0110.0450.0250.0050.8821.0000.0090.027 Pycnococcus provasoli RCC2440.0800.0170.0610.3050.0240.0060.0190.0310.0090.0020.0210.9711.0000.0130.019 Pseudoscoufieldia marina RCC2610.0710.0060.0570.2870.0360.0180.0250.0780.0030.0230.8711.0000.0060.027 22 pigments and 30 strains

7. BLA148/9 BLA78/7 RCC287 BLA82/7 RCC343 (Ostreococcus IV) RCC116 (O. tauri) RCC344 (Ostreococcus IV) RCC371 (Ostreococcus IV) RCC356 (Ostreococcus IV) BLA122 RCC393 (Ostreococcus II) RCC136 (Prasinococcus) RCC137 (Prasinoderma) Chlorodendrales Mamiellales Prasinococcales

8. Clade II Mamiellales Clade V Pseudoscourfieldales (Pycnococcaceae) Clade VI Prasinococcales Clade III. Pseudoscourfieldales (Nephroselmidaceae) Clade I. Pyramimonadales Clade VII Clade IV. Chlorodendrales

9. A simple pigment key to assign Prasinophyceae strains to the 7 clades Other species from Clade III have Siphonaxanthin or Loroxanthin or Loroxanthin derivatives.

10. Final Summary The two types of pigmentation in Prasinophyceae according to Egeland et al (1995) were clearly identified by the cluster analysis In addition, the clones could be grouped into the three established orders. However, the pigment data also point to a large discrepancy within the order Chlorodendrales (new order?) There were important non-common features within each of these two groups. Perhaps the most remarkable were the following: Presence of Chl c3 in two clones (BLA122, RCC393) Simultaneous presence of Prasinoxanthin and Lutein in two clones (RCC136, RCC137) Presence of MgDVP in a Chlorophyceae-type prasinophyceae (BLA78/7) Absence of Violaxanthin in one clone (RCC137) It's the rule

11. Chl c 3 : Chl c CS-170 or MVChl c 3 or both?

13. Summary. Pigments of the prasinoxanthin-containing Prasinophyceae are very useful to assign those strains to the three clades identified by SSUrDNA analysis: clade II, V and VI. Pigments of the prasinoxanthin-less Prasinophyceae provided a much less clear picture of the taxonomic affinities of those strains. The new clade VII contained the pigment composition more similar to Chlorophyceae among the strains studied. A pigment key is proposed to assign strains to the 7 Prasinophyceae clades. However, data from literature imply that Clade III (Pyramimonadales) may confound this classification. A MVChl c 3 -like pigment was present in strains isolated (or related) from the deep euphotic zone. Pigments have been proven a useful but no definitive tool for taxonomic work within the class level.