[Note: these are rough minutes taken down during the meeting itself. They are intended for archival purposes; please excuse the rough edges -- we would appreciate getting feedback from anyone about errors or omissions in this document.] [revised Nov. 3, 1995] USDA Green Plant Phylogeny Research Coordination Group Workshop #2 in Association with the Annual Meeting of the Phycological Society of America 11-12 August 1995 Breckenridge, Colorado Attendees: Mark A. Buchheim, PI (MAB), Russell L. Chapman, Co-PI (RLC), William Birky (WB), Charles Delwiche (CD), Marvin Fawley (MF), Thomas Friedl (TF), Paul Fuerst (PF), Alan Gaspar (AG), Thomas S. Kantz (TK), Louise Lewis (LL), Juan M. Lopez-Bautista (JLB), Rick M. McCourt (RMC), Susan T. Meiers (STM), Jeanine Olsen (JO), Karen Phillips (KP), George T. Tziahanas (GT), Wytze Stam (WS), Frederick "Rick" Zechman (RZ) Session 1 - Friday 11 August, 8:30 a.m. The meeting began with a general introduction of the participants and brief comments on business details (i.e., the workshop schedule, the procedures for travel reimbursements, etc.). The possibility of having an evening session was approved with the understanding that we would probably still have Saturday morning session. MAB presented the historical background of the grant (and fact that there was a joint program among the NSF, USDA, and DOE) and brief review of the goals of the current workshop. It was noted that the goals for the grant were: 1) coordination, 2) development of mechanisms for data base management (maintaining a data availability matrices, etc.) 3) dissemination of information of group activities (Brian Speer at UCB has developed the page it will be posted via the Algal List Serve and the PSA List Serve,; our group page could be found via the UCB home page WWW.Berkeley.edu; Michael Sanderson's Tree Base and the Maddison brothers' Tree of Life [the latter will be demonstrated later in this workshop]), 4) stimulation of new approaches (especially data analysis and handling of large data sets), and 5) stimulation of collaboration. In the discussion of the history of the proposal and the grant, it was noted that the focus is on the green algae and lower land plants. MAB reported on the workshop at Berkeley and described the funding mechanisms. The original grant proposal had only 30 or so direct participants, but even the first workshop exceeded that number. The goal is to develop lists of exemplar taxa and develop a list of taxonomic characters that should be included in the "grand analysis" of green plant phylogeny. An ultimate goal of this project is some sort of book with chapters on each major taxon group and a grand analysis on CD ROM. The proceeds from this project would go into an endowment for green plant phylogeny research. Everything included in this book would be published data. There seems to be strong interest in the project from publishers (e.g. from Academic Press) and there is reason to believe that other publishers like Sinauer would be interested. N.B. the minutes from the first workshop (cf. last page of the minutes) cite several possible publishers. The data availability matrix will serve as a mechanism for coordinating activity. RLC commented on the suggestion for a database of DNA samples might be put of the web. CD indicated that there was also consideration of posting a database of primers used. MAB discussed the tentative schedule for the current workshop: the groups of green algae to be covered and the character selection information from the Berkeley meeting. MAB asked if participants wanted to discuss the goals of the group. He noted that there had been some discussion of proprietary and non-proprietary databases. The general consensus at the Berkeley meeting was to create only a non-proprietary database. CD mentioned the interest in having a depository for alignments. MAB asked if Tree Base would meet this need. There was a discussion of Tree Base and it was noted that Michael Sanderson has developed Tree Base with Michael Donoghue. Sanderson is now at University of California, Davis. CD noted that there are some special tools associated with Tree Base that will be helpful. JO commented on the possible uses of the databases etc. WB raised the question of how to handle changes in alignment. There was a general discussion of how such databases would be handled and how the book would deal with conflicting alignments etc. It was noted that the book would have to have an editor or editors who would deal with some of the questions raised. MAB discussed participants. RLC discussed the five levels of participation: the PIs, the proposal participants, the workshops participants, the mailing list members, the web readers. MAB discussed others who were invited but were unable to attend: Charles O'Kelly (Chlorophyta in general), Ĝjvind Moestrup (Prasinophytes), Michael Melkonian (Chlorophyta in general), Jeremy D. Pickett-Heaps (Chlorophyta in general), Mattox and Stewart (Chlorophyta in general), Gary Floyd (Chlorophyta in general), Volker Huss (all green plants), Dabashish (Prasinophytes). Additional potential participants were listed in the project: Bob Price for conifers, YQ for angiosperms, Cliff Morden, Richard Pienaar (Prasinophytes), Stuart Sym (Prasinophytes), Peter Vroom and Cecilia Smith (ulvophytes), Geoff Wolcott, Hanya Hanszka (Coleochaete), Martine Closas (paleo.), WS (marine algae), Erin Kaster, Rhynie (paleo), and Monique Feist (Charales, paleo). CD raised the issue of future meeting plans: (it was noted that it is clear that not all interested parties can participate in all workshops): next February 15th-20th in Baton Rouge, LA; RLC reported on plans for the data analysis workshop; also in 7-13 July 1996 International Charophyte symposium in Madison might be appropriate site for a workshop; also in 1996 global characters and morphology at Seattle meeting of AIBS; JO mentioned various algae- oriented meeting (e.g., International Phycological Congress in 1997); 1997 in Montreal the fern group and the bryophyte groups; 1998 fossil; 1999 whole group meets in St. Louis at the International Botanical Congress. There was some discussion of the need for the "algal-people" to meet on at least two more occasion. Therefore, perhaps an algal meeting in 1998 would be useful and important. There was a discussion of the need for taxon sampling in the chlorophycean sensu lato lineage. WB noted how different the situation is for algae vis-a-vis vertebrates or embryophytes; therefore, taxon sampling is a difficult topic. JO agreed and noted the specific example of Cladophora and the need for many representatives to cover this taxon. WB suggested that the whole question of taxon sampling for green algae will have to be an ongoing, self-correcting process because we cannot know at the start which are the best exemplar taxa. CD asked whether there were ongoing broad surveys of green algal diversity. MF noted that he had submitted a proposal for such a survey. There was a general discussion of this kind of research on algae and protists (e.g., the work of Lisa Campbell). MAB reported on the criteria used at the Berkeley workshop: 1) availability, 2) nomenclatural types, 3) model systems, 4) Basal and/or isolated taxa, 5) "broad" sampling of diverse taxa, and 6) anomalous taxa Break at 12:00 noon Session 2 - 1:30 p.m. MAB discussed the NEXUS format and turned the discussion over to CD who presented the results of the Charophyte subgroup at the Berkeley meeting. CHAROPHYCEAE CD noted that the Trentepohliales, Stichoccocus, and Raphidonema are not charophycean based rbcL analyses, RLC said the same is true in rRNA analyses. Exemplar Taxa: Chara connivens Nitella sp. Tolypella sp. 1 Tolypella sp. 2 Chlorokybus atmophyticus Klebsormidium laccidum Klebsormidium sp. 1706 Coleochaete orbicularis Coleochaete scutata Coleochaete soluta Coleochaete conchata Coleochaete pulvinata Coleochaete rregular Coleochaete nitellarum Coleochaete circularis Chaetotheke sp. Chaetosphaeridium ovalis Closterium sp. Gonatozygon sp. Penium sp. Cosmarium sp. Micrasterias sp. Spirogyra maxima Spirogyra grevilleana Zygnema peliosporum Zygnemopsis sp. Mougeotia sp. Mesotaenium sp. Cylindrocystis sp. Netrium sp. Spirotaenia sp. Cosmocladium sp. Onychonema sp. Entransia sp. 1) Charales: Need a Chara and a Nitella to guard against long branch; no need for Lamprothamnium (probably a Chara); Nitella better than Tolypella (the latter is less abundant world wide); using a specific species of each (one than is clearly and accurately identified); we could get good recommendation from Vernon Proctor for each (incl. ones that are good to grow [e.g. C. connivens], and perennial); N. flexilis, N. axillaris (RLC and Mark Ragan sequenced) and N. sp. (German group), N. opaca 2) Chlorokybus atmophyticus (hard to get DNA from it in Chuck Delwiche's lab; worked OK for Wilcox & Chapman's lab) only collected from nature twice 3) Klebsormidiales: Need to know more about the group. [edit note: Hans Sluiman of the Royal Botanical in Edinburgh is now working on these algae.] Klebsormidium flaccidum or second taxon in the group Re: Stichococcus bacillaris and Raphidonema longiseta TF and RLC agree Raphidonema longiseta is not Charophyceaen 4) Coleochaetales Coleochaete scutata (several cultures available) first choice: Coleochaete orbicularis (not a good name) Linda Graham monoecious these two species are close (corticated zygotes) second choice: Chaetospheridium ovalis hard to grow CD has and will release Chaetospheridium globosa third choice: no data for the other group Coleochaete circularis group (free zygotes loosely covered) difficult grow, CD would release his culture when he published his rubisco paper fourth choice: Coleochaete nitellarum grows well (grows on Nitella) Coleochaete DISC. studying the Coleochaetales would be very useful and could lead to polarization of morphological. characters; CD will be working on this Chaetotheca good to study if CD can find, never cultured, never studied; CD will study Awadhiella indica (named by Indian group) like a Coleochaete circularis 5) Zygnematales (RMC) placcoderms are monophyletic saccoderms etc. confused Marion Mix's four families are good first priority: Micrasterias (model system) second priority: Gonatozygoacae: Gonatozygon or Genicularis third priority: Closteriacae: Closterium acerosum Cosmocladum (colonial) Onchychonema fourth priority: ? fifth priority: Peniaceae: Penium Desmidiaceae: Cosmarium botrytis Michael Melkonian will come out with information on nine taxa in a PNAS paper Mesotaeniaceae (not monophyletic) Barbara Surek first priority: Spirogyra maxima Sirogonium sister taxon to S. maxima Mougeotia scalaris (used for phytochrome work) second priority: Mesotaenium Spirotaenia Zygnema peliosporum? Cylindrocystis third priority: Zygneopsis (like Zygnema) fourth priority: Netrium often basal (in different places) 1.add Tolypella (perhaps T. porteri) RMC has ct. sequence for three other species representing two sections T. intricata T. nifida (these represent the two subgroups) 2. 5-6 Klebsormidium spp. in culture collections? 3.RLC check culture collections for all: species in the Klebsormidiales and Coleochaetales (RMC McCourt has info. on all of the Zygnematales) Strains Available (WFCC - MIRCEM World Data Center on Microorganisms) 23 October 1995 1. Klebsormidium flaccidum 2. Klebsormidium marinum 3. Klebsormidium spp. 4. Klebsormidium sterile 5. Klebsormidium subtilissimum 6. Klebsormidium subtilissium 7. Ulothrix confervicola 8. Ulothrix crenulata 9. Ulothrix fimbriata 10. Ulothrix gigas 11. Ulothrix minuta 12. Ulothrix sp. 13. Ulothrix trentonense 1. Coleochaete nitellarum 2. Coleochaete scutata 4.VH: they sequenced Coleochaete nitellarum on a Chara sample by mistake 5.CD & LG: add Coleochaete pulvinata LG has morphological. data, CD has rbcL, this one is in the orbicularis group), C. soluta and C. conchata are similar; C. irregularis would be good addition. Still need one from circularis group c. circularis (most basal) c. irregularis c. pulvinata + nitellarum (most derived) c. nitellarum (sister to pulvinata) 6.CD: wants to look at Chaetospheridium ovalis (video photomicroscopy) of oogamous reproduction with initial egg release then sperm release and fertilization of free zygote. 7.therefore total Coleochaetales = 9 plus Chaetotheke might be appropriate if observations on sexual reproduction show that it is different (it is a Myriophyllum epiphyte) and rather common) Also, Moestrup may have another Chaetospheridium 8.Zygnematales: 54 genera, 3 families, Zygnemataceae 12 genera Mesotaeniaceae 6 Peniaceae 2 Gonatozygaceae 2 Closteriaceae 3 Desmidiacae 30 +/- Total Zygnematales: 54 sample 16 (incl. Entransia) 9. anymore Chlorokybus sp. or isolates? 10. Klebsormidiales 2 or more Coleochaetales 9 and maybe Chlorotheke Zygnematales 16 Chlorokybales 1 Charales 3 There was a general discussion of the taxa selected and the fact that in the context of a broad survey ca. 34 taxa would give adequate coverage for these algae. WB noted that in some cases one can completely sequence a gene for one taxon and then only sequence a small region of the molecule for related species. There was a discussion of the rationale behind the selection of taxa for Coleochaete as one example of the charophycean exemplar selections. It was noted that the Zygnematales includes much of the diversity of charophycean algae, but the broad survey does not require more taxon sampling. JO raised the question of the geographical origins of the exemplar taxa and there was a general discussion of the amount of taxon collecting from South America, Africa, etc. Charophycean Exemplar Taxa Chara connivens Nitella sp. Tolypella sp. Tolypella sp. Chlorokybus atmophyticus Klebsormidium flaccidum Klebsormidium sp. 1706 Coleochaete orbicularis Coleochaete scutata Coleochaete soluta Coleochaete conchata Coleochaete pulvinata Coleochaete irregularis Coleochaete nitellarum Coleochaete circularis Chaetotheke sp. Chaetosphaeridium ovalis Closterium sp. Gonatozygon sp. Penium sp. Cosmarium sp. Micrasterias sp. Spirogyra maxima Spirogyra grevilleana Zygnema peliosporum Zygnemopsis sp. Mougeotia sp. Mesotaenium sp. Cylindrocystis sp. Netrium sp. Spirotaenia sp. Cosmocladium sp. Onychonema sp. Entransia sp. MICROMONADS Exemplar taxa for the micromonads were discussed. MF indicated that Tazmanities should be added and that Pterosperma was important. Pyramimonas is important. Mantoniella is important and available. Pedinomonas minor (freshwater and marine) there are three subgenera and some for which much electron microscopy have been done, MF and TK were asked to prepare a list of appropriate taxa, MF noted that the Pseudoscourfieldia in Bob Anderson's collection CMMPSTRAIN strain is NOT Pseudoscourfieldia, the real one is available from Franzen (Oslo), Resultor, Scourfieldia (at SAG), Mamiella gilva (available from Plymouth) is very close to Mantoniella, therefore may not need to have all of them. Bathycoccus (close to Mamiella, Micromonas, and Mantoniella) and Picnococcus (close to Pseudoscourfieldia). There are three coccoid clones unnamed strains from IVE 5G, URI 266G, and IID 15A should be examined and are available. TF noted that there is a collaboration between Michael Melkonian,Volker Huss, and Inoue and they are publishing a paper on these. Mesostigma (available from SAG). Nephroselmis olivacea Micromonas pusilla Mantoniella squamata Mesostigma viridis Pterosperma Tasmanites Pedinomonas sp. 1 Pedinomonas sp. 2 Scourfieldia sp. Pseudoscourfieldia marina Pyramimonas subgenus 1 Pyramimonas subgenus 2 Pyramimonas subgenus3 Mamiella gilva Bathycoccus sp. Pycnococcus sp. IVE5G (coccoid) URI266G (coccoid) IID15A (coccoid Scherffelia dubia Tetraselmis striata Cymbomonas sp. ULVOPHYCEAE JO presented some exemplar taxa for the Ulvophyceae following a discussion with some of the other participant. Ca. 100 taxa are needed. Cladophorales, Siphoncladales ca. 20 genera Acrosiphonales, Ulvales, Ulotrichales ca. 20 genera Dasycladales ca. would like to add two more (which are basal ) ca. 11 genera Caulerpales (Derbesia- Halicystis group, Codium, [Paul Silva trying to get funding for work on this genus which is difficult because other organisms live within the alga; Bryopsis - Inoue group in Japan has one sequence, Erin Caister is planning to work on Bryopsis at Duke, JO and RZ have partial sequences); need good sampling of Caulerpa; Udotian taxa should also be examined and one species per genus is probably OK except for Halimedia which may require heavier taxon sampling; several other genera would require only one exemplar; because these are very ancient tropical taxa they require extensive sampling. JO reviewed a number of miscellaneous taxa in the group. Freshwater forms are needed and the participants did not know too many investigators working on these algae other than Linda and Jim Graham. The Trentepohliales are within this group and only a few exemplars are needed (viz., Cephaleuros virescens and Trentepohlia spp. [perhaps both the lichenized form available via the UTEX collection and a named species.) There was a general discussion of whether or not the total of ca. 100 needed taxa could be reduced. JO indicated that there could be some indication of priority taxa, but the Caulerpales require extensive sampling and the number needed cannot be reduced much. RZ noted that there is tremendous genetic diversity within the Caulerpales and within the Ulvophyceae (which may not be monophyletic). The Caulerpales are the least well known among the chlorophycean greens (they are difficult to grow in culture, they may exhibit a lot of morphological convergence, there are epiphytes and internal endophytes or endozooa, interstitial contaminants. There was a general discussion of the level of sampling needed. The broad picture of chlorophycean phylogeny may require extensive sampling just to get a clear picture of where parts of the Ulvophyceae belong. WB noted the fact that much of our phylogenetic research has been focussed on the "popular" organisms (e.g. birds, vascular plants). LL commented on the fact that the inclusion of numerous ulvophyceae algae would indicate the interesting questions etc. RM commented on the fact that for the streptophytes with 200+ taxa and many, many workers it is not clear that all questions would be answered. CD noted that for the green algae we may not need or want massive data sets for all the taxa. It was agreed that the ulvophyte group of investigators would work on the master list and attempt to suggest priorities. TREBOUXIOPHYCEAE Trebouxia magna Myrmecia israelensis Myrmecia bisecta Dictyocloropsis reticulata Microthamnion kuetzingianum Fusochloris perforatea Chlorella saccharophila C. luteoviridis C. vulgaris Protheca wickerhamii Cocomyxa sp. Nannochloris sp. Chlorella mirablis C. ellipsoidea Stichococcus bacillaris Apatoccus lobatus Eremosphaera viridis Golenkinia minutissima Pleurastrum terrestre Leptosira There was a general discussion of whether or not this list was adequate and it was felt that it was generally enough. There was a discussion of the fact that there are multiple lineages of Chlorella spp. and some will be in the Chlorophyceae CHLOROPHYCEAE 30 chlamydomonadalean taxa Chlamydomonas reinhardtii Chlamydomonas mexicana Chlamydomonas moewusii Haematococcus lacustris Haematococcus zimbabwiensis Stephanosphaera pluvialis Chlamydomonas monoica Chlamydomonas moewusii Chlorogonium elongatum Dunaliella parva Asteromonas gracilis Brachiomonas submarina Polytoma uvella Polytoma 62-27 Polytomella spp. Carteria eugametos Carteria radiosa Spermatozopsis similis Astrephomene guber. Gonium pectorale Eudorina elegans Pleodorina californica Volvox aureus ___ Sphaeroplea annulina Microspora sp. Radiofilum transversale Cylindrocapsa geminella Geminella minor Radiofilum conjunctivum ___ Pleurastrum insigne Hormotilopsis tetravac. Planophila terrestris Hormotila blennista Chlorella fusca Scenedesmus obliquus Chlorococcum ellipsoid. Hydrodictyon reticulatum Pediastrum duplex Neochloris aquatica Characium hindakii Ankistrodesmus falcatus Monoraphidium griffithii Bracteacoccus minor Interfilum sp. Gloeotila sp. Chlorosarcina sp. ___ Chaetophora crassa Draparnaldia plumosa Uronema belkae Fritschiella tuberosa Stigeoclonium Thamniochaete Chaetonema Aphanochaete Schizomeris leibleinii Pseudoschizomeris caudata ___ Oedogonium cardiacum Oedocladium carolinianum Bulbochaete hiloensis ___ Chaetopeltis Gloeococcus Paulschulzia pseudovolvox Tetraspora lubrica Palmella texensis ___ Gongrosira papillatum Chlorochytrium lemnae Coelastrum microsporum Tetradesmus cumricus Diplosphaera chodatii Dictyosphaerium ehrenbergianum Selenastrum capricornutum Kirchneriella cornata Heterochlamydomonas sp. Dictyochloris sp. Heterotetracystis sp. Fasiculochloris sp. It was noted by TF that the Volvox carteri sequence from Rausch et al. may be wrong (they never looked at the alga). PF said they had a full sequence (SSU rRNA) and it is very different from Volvox carteri. WB said there are probably two clades of Polytoma spp. Tetraspora Binuclearia etc. get list from MAB Cylindrocapsa involuta in misc. ulvophyceae Chlorococcales (including Chlorosarcinales) ca. add: Ankistrodesmus falcatus Monoraphidium braunii Bracteaccocus minor Interfillium, glaeotila, chlorasarana Chaetophorales: add: Schizomeris leibleinii Pseudochizomeris caudata Chaetophora crassa Aphanochaete use type species Oedogoniales Oedo gonium cardiacum, Oedocladium carolinianum and Bulbodialte misc. Gongrosira papuasica etc. get list from MAB RMM gave a demonstration of the Tree of Life page and program developed by the Maddison brothers on the web. He demonstrated on a page could be made for the green algae and land plants. He invited everyone to consider making a contribution to the Tree of Life and send appropriate files to the Maddison brothers. He indicated that participation is welcome. For teaching, people can make and use a local version (i.e., one that is not published on the Web). http://wwwphylogeny.arizona.edu/tree/life.html MAB stated that we needed to discuss some additional miscellaneous taxa, discuss characters, and that if we wanted to cooperate in the 1997 International Phycological Congress we would need to act now since the planning committee is meeting next week. The possibilities for participation include: a symposium or a workshop or both. These activities would let phycologists from around the world know what is going on. We would need the title and co-conveners. There would be a local co-convener (e.g. JO and WS) and at least one of the PIs. There was some discussion of the possible content for a workshop or symposium. It was agreed it must be more than a progress report. The symposium/workshop would have to compete with others for the ca. 16 to slots for symposia. Some update on the WWW pages etc. and Tree of Life would help spread the word. One paper could describe how the algae are being handled in the Green Plant Phylogeny project. WB asked whether a title based on International Communications etc. would be appropriate. MF suggested that a presentation on the project combined with a workshop would be good. The possibility of a plenary lecture was mentioned. There was a discussion of the types of symposia that would be considered by the International Phycological Congress Organizing Committee. There was some agreement that a symposium that dealt various groups of algae (greens, reds, etc.) would be good, but the possibility of focussing just on the international green algae part of the Green Plant Phylogeny Project. Of these two possibilities, there was general support in favor of not limiting a symposium to just the greens, but perhaps focussed on the processes and the different groups. There were comments on the various ways theproject grant could support participation of faculty and graduate students at the International Phycological Congress (10-17 August 1997). The 1998 PSA Meeting will be in Baltimore (with AIBS) or Flagstaff. TF indicated that in Germany that there was a loose organization of several labs looking at green algae and chromophytes, as well as land plants. Break at 5:00 p.m. Session 3 - Evening Session 7:00 p.m. INCERTAE SEDIS Prasiola crispa Prasiola stipitata Ostreobium quecketti (does have chls a & b) Hafniomonas Chlorella spp. Diplochaete solitaria (cf. Michael Wynne and comments about diatoms: seems to be a diatom!) Session 3 - 7:00 p.m. OUTGROUPS The selection of the outgroup depends in part on the specific data being analyzed (e.g., if you are looking at a chloroplast gene like rbcL, the selection of the outgroup might be different from that chosen in an 18s rRNA analysis. The chlorophycean lineage would probably serve as an outgroup to the streptophytes. How far out should the outgroup be? Are reds (Rhodophyta) and browns (Phaeophyta) too far out? Are some of the amoeba groups and the Cryptomonads appropriate? How many different outgroups should be used. If a variety of very different outgroup taxa yield the same topology for the ingroup, the outgroup selection should not matter. This would be an appropriate topic for the data analysis workshop. CHARACTERS Genes used for phylogenetic studies in higher plants: rbcL: Highest priority for higher plants and also for algae cp 16S; problems with bacterial contamination cp introns co 23S: MB good gene to start working, Monique Feist. cp ndhF: high priority cp atpB: high priority cp matK nu 18S: highest priority for algae nu 26S nu phytochr nu heat sho nu centrin nu VATP "leafy": totally unacceptable cp/nu tufA: carries the problem with bacterial contamination. cp ribosom cp psA cytokinetic Post Zygoti nu introns MAB presented some of the comments from the Berkeley meeting. There was general agreement that rbcL and nuclear- encoded SSU rRNA were the highest priority molecular data to be gathered. LL noted that rbcL sequences are almost too divergent for work with the bryophytes. The second priority for a chloroplast gene was discussed. The possibilities of prokaryote contamination makes 16S rRNA genes less useful than rbcL. CD said that there are some problems with using tuf A genes for such a study. There was a discussion of the utility of the rbcL gene and the report by Jim Manhart et al. on the results. There was a question about how many rbcL sequences were available for non-charophycean algae. RMM has developed some ndhf gene sequences and would share the primer information. As potentially useful characters ct atpB and ndhf were ranked next. JO asked if any total ct genome sequencing projects were underway for a cross-section of taxa. No one was familiar with a project. MAB asked if anyone knew what Monique Turmel and Claude Lemieux planned for future sequencing. RLC asked if any of the molecular characters listed by the Berkeley workshop were totally unacceptable. JO asked CD about his opinion on tuf A. For both nuclear-encoded and ct-encoded LSU there is not much data available. WB noted that the different domains with different degrees of variability in LSU are useful, but the lack of data is a good reason for not focussing on LSU. PF noted that the presence introns (that can make sequencing difficult) in the LSU makes it questionable to suggest sequencing this large molecule. JO said that complete data sets for two or three genes would be better than a scattered incomplete set of different genes. There was general agreement that three complete molecular data sets would be fantastic and that SSU rRNA (nuclear-encoded) and rbcL (ct-encoded) would be the first two. There was a discussion of the what the third gene should be. TK suggested that a nuclear-encoded gene should be preferred over a chloroplast gene. This suggestion was generally supported, but the problem of parallogy in some nuclear-encoded genes was discussed. Heat shock proteins and actin proteins were mentioned and briefly discussed. The parallogy of actin genes in land plants would not be a problem for the algae. TK noted that for rooting the green plant tree a nuclear-encoded gene would be better than a plastid gene. PF questioned whether or not, mitochondrial genes would be useful in the rooting. WB noted some additional problems with mitochondrial genes; therefore, using a nuclear-coded gene would be better as the third gene. CD stated that it appears that there is a need to determine a nuclear-encoded protein gene that would be useful. RMM said that some may be working on this and we have an answer. Liz Waters (formerly at University of Arizona) was looking into this. JO posed the question of gathering a set of DNAs from a variety of green algae and have a commercial firm sequence several of the potentially useful genes. The discussion returned to heat shock proteins and vacuolar ATPases. Heat shock proteins may be a poor choice because of parallogy. Carol Bult at TIGR might be able to provide information. RMM doing a survey of ct genes: ndhf , mat K, and atp B, etc. for the Charales. He is using Olmstead's primers. TF noted that they are looking at the actin gene. They have looked at some Trebouxia spp. and are working on primers for other algae. He thinks they will have some more done within six months. WB asked why the Berkeley group came up with so many genes? CD the idea was not to do all of the genes, but the objective was to get about 10,000 kB of sequence to get robust resolution. WB noted that are fewer phycologists and less money for this research and therefore we should avoid diluting our resources by going after too many genes. LL raised the question the question of which actin algal genes had been sequenced and TF noted there were two charophytes. The potential problem of actin parallogous genes was mentioned again. The point at which duplication of the actin gene occurred would itself be interesting and useful. RZ noted that we need to look at the amount of sequence divergence between very distantly related green algae. The final consensus was that there is a good possibility that there are several candidates for the needed third gene (among nuclear-encoded protein genes), but additional research clearly is needed. There was a brief discussion of the topics to be considered tomorrow and the preparation of the ulvophyte list. Adjourned 8:50 p.m. Session 4 - Saturday 12 August 1995, 9:00 a.m. NON-MOLECULAR DATA Major classes of characters: cytokinetic apparatus; post zygotic events; mitotic apparatus; flagellar apparatus; pyrenoid; life history; gross morphology; ultrastructural features; vegetative and reproductive cell surface structures; wall structure and composition; pigments; plastid number and morphology; biochemistry/physiology; cytoskeleton; mitochondrial morphology RLC noted the need to assign "homework" to generate specific lists of characters and character states for each character suite. WB will review mitochondria and chloroplast inheritance (post zygotic events) RMM and CD Charophytes TF Trebouxiophyceae JO, WS, and RZ Ulvophyceae LL Chlorophyceae MF and TK Micromonads Pigments MF Flagellar apparatus PF and Gary Floyd Pyrenoid: TK Life History: skip Gross Morphology: skip cells surface structure: TK chloroplast: skip biochemistry and physiology: skip cytoskeleton: skip Melkonian (JO will ask him) mitochondria morphology: skip MAB noted that the MoBot paper might be helpful. RMM noted that with this information in the minutes, people could contact one another about specific questions. RLC noted that participants should feel free to contact other phycologists and solicit their help. DEADLINE: 30 September 1995 - Verify Confirm Exemplar taxa list and list of characters and character states clearly defined, also indicate availability of molecular data for those taxa. As soon as possible, MAB and RLC will send out a consensus character and character state list. Then everyone will review the draft. It was agreed that we should begin developing the data bases of: Cultures available, primers available, sequencing in progress, research plans. It was agreed that everyone would email the information email to MAB. CD noted that we could set up a moderated list serve for this group. TF suggested the preparation of a form and MAB agreed to prepare a format and will email it out. It was noted that making cultures available could be a problem if there are too many requests. RLC noted that there is no reason to put a culture on this list if it will not be released, and that if an alga is not readily available, one should reconsider whether that taxon should be an exemplar. CD said that availability was only one criterion and not necessarily the most important criterion. RLC stated LEIDEN:"Global Consortium in Algal Evolution: Partnerships, Process, and Progress" Co-conveners: MAB and Volker Huss Participants: MAB, (RLC), Volker Huss, Gary Saunders, Wilson Freshwater, Mark Ragan, Suzzane Frederique, Hiroshi Kawai (Ian Tam), Akira? Peters, for the chromophytes: introductory talk on the Partnerships, Process, and Progress; 1 talk from the red project; brown group; (Huss or Inoue, Friedl: coccoid green group German/Japan) Goals: to filling the data matrix to create an international group of green algal phycologists to reunite red, green, chromophyte, and brown algal research. Progress reports Logistic for the group It was noted that the International Phycology Congress would provide a great opportunity for presenting information on what we are doing and how we are doing (collaborative research). JO and WB noted that collaborative research is an important issue in Europe and the US. JO described some of the possibilities for a symposium and the mechanisms for the handling the schedule etc. WB discussed the various aspects of this project that could be presented at a symposium (3 talks on the process, politics, and progress plus a fourth talk on other initiatives). MF agreed that this structure would be good. There was a general agreement that a workshop would be desirable with or without the symposium, and that one full day is needed. The workshop would be an update on the Green Plant Phylogeny Project - the Algal Component (with ca. 20 participants by invitation). The organizing committee will be meeting next Friday the 18th of August. PF commented on the possibility of submitting a proposal for funds to help. MISC. MAB will send the exemplar taxon list; the shell of a information on cultures available, primers available, planned research, and sequencing in progress; and the email address list. Participants will send their list of characters and character states (with definitions of character states as necessary). TF Reisser at ASAG collection working on it; Linnaeus program phytoplankton ETI; TF and LL will send list of who is doing what (TF, LL, SW, Takishi Yakanama (Inoue), PF and Gary Floyd, Volker Huss; "Schwerpunkt" grant for studies on the Molecular Evolution in Plants (incl. green plants and chromophytes; there are many people involved, incl, Linda Medlin and other diatomists; organizers: Uwe Jensen and Hurka) Rüdiger Cerff more or less in charge of the algae activities and is involved in the International PHycologicalCongr ess. There may a Japanese brown algae group with group funding. JO said there maybe a large diatom consortium but there was no other information available. TF will send RLC a short report on this workshop and RLC will share it with RMM and then send a revised version to MAB and BDM, then the approved version will be sent to all participants (incl. the land plant people). RLC raised the question of other activities this Project should support to meet its goals. JO suggested aggressive "advertising" of the Project. RZ asked bout Public Radio. RMM noted that the UCB public relations office is supposed to publicize the Project. CD suggested that a List Serve or Moderated News Group for this group (i.e., the algae people). RMM commented on the Tree of Life and the fact that he is preparing a letter to invite people to look at what he has done and to contribute to the algal page in the Tree of Life. JO asked about plans to report on this group to the PSA. TK noted that RMM would put a note in the Newsletter. It was noted that we would have some informal gathering at the PSA meeting in 1996 and that we have plans for the International Phycological Congress in 1997. CD noted that 1998 would be a good year to do something formally in the PSA meeting (perhaps a symposium followed by a workshop). JO noted that there is some time between the PSA and the EPS, so maybe something could be planned. It was agreed that there would be an informal meeting of this group at the PSA meeting in Santa Cruz. Adjourned 12:05 p.m.