Logarithmically rising hh and $gamma h$ cross-sections some features of model versus experi
A Single Species, Micromonas pusilla (Prasinophyceae), Dominates the Eukaryotic Picoplankton in the

A PPLIED AND E NVIRONMENTAL M ICROBIOLOGY,July2004,p.4064–4072Vol.70,No.7 0099-2240/04/$08.00ϩ0DOI:10.1128/AEM.70.7.4064–4072.2004Copyright©2004,American Society for Microbiology.All Rights Reserved.A Single Species,Micromonas pusilla(Prasinophyceae),Dominates theEukaryotic Picoplankton in the Western English ChannelFabrice Not,1Mikel Latasa,2Dominique Marie,1Thierry Cariou,1Daniel Vaulot,1*and Nathalie Simon1Station Biologique,UMR7127CNRS,INSU and Universite´Pierre et Marie Curie,29680Roscoff,France,1and Institut de Cie`ncies del Mar(C.S.I.C.),08003Barcelona,Spain2Received17September2003/Accepted11March2004The class Prasinophyceae(Chlorophyta)contains several photosynthetic picoeukaryotic species describedfrom cultured isolates.The ecology of these organisms and their contributions to the picoeukaryotic communityin aquatic ecosystems have received little consideration.We have designed and tested eight new18S ribosomalDNA oligonucleotide probes specific for different Prasinophyceae clades,genera,and ingfluorescentin situ hybridization associated with tyramide signal amplification,these probes,along with more generalprobes,have been applied to samples from a marine coastal site off Roscoff(France)collected every2weeksbetween July2000and September2001.The abundance of eukaryotic picoplankton remained high(>103cellsml؊1)during the sampling period,with maxima in summer(up to2؋104cells ml؊1),and a single green algalspecies,Micromonas pusilla(Prasinophyceae),dominated the community all year round.Members of the orderPrasinococcales and the species Bathycoccus prasinos(Mamiellales)displayed sporadic occurrences,while theabundances of all other Prasinophyceae groups targeted remained negligible.Several studies have demonstrated the importance of eukaryotic picoplankton(cell size,0.2-to3-m)in terms of biomass and productivity in the euphotic zone of oceanic oligo-trophic waters(15),as well as in coastal waters(10).To date,only ϳ40species belonging to nine algal classes(Chlorophyceae,Pra-sinophyceae,Trebouxiophyceae,Prymnesiophyceae,Bolido-phyceae,Eustigmatophyceae,Pinguiophyceae,Bacillariophyceae, and Pelagophyceae)of photosynthetic picoplanktonic eukaryotes have been formerly described(41).However,phylogenetic anal-yses of sequences retrieved from natural samples in different oceanic regions have demonstrated much higher diversity,since many of these sequences do not correspond to any described taxa (19).The contributions of the different taxonomic groups to the picoplanktonic biomass,diversity,and ecology are poorly known because simple and reliable methods to detect and quantify such organisms in natural samples are lacking.Pigment signatures, scanning electron microscopy,and serial dilution cultures suggest that the classes Prasinophyceae(division Chlorophyta),Pelago-phyceae(division Heterokontophyta),and Prymnesiophyceae are major components of the picoplankton biomass in different ma-rine systems(20,35).Among these,the class Prasinophyceae contains several photosynthetic picoeukaryote species.This class is considered to be the most primitive in the green lineage and to have given rise to all other green algal classes,as well as to the land plants (34).Members are known to be common in temperate and cold regions and can occur as prominent constituents of ma-rine picoplankton(38).Within these organisms,genera such as Ostreococcus,Bathycoccus,and Micromonas have been de-scribed in coastal waters(4b,6).Micromonas pusilla(the only described species in the genus Micromonas)has been identified as a major component of the picoplanktonic community in several oceanic and coastal regions,such as the Mediterranean Sea(39),the Norwegian Sea(37),and central California wa-ters(35).However,the techniques used to establish these facts (microscopic identification of cells presenting few morpholog-ical characteristics or serial dilution cultures)are time-consum-ing and incompatible with extensive ecological studies.In con-sequence,the precise distributions and the seasonal dynamics of an apparently very common picoplankter,such as M.pusilla, are poorly known.The aim of this work was to identify and study the seasonal variations of the dominant taxa in the picoeukaryote commu-nity at a coastal site of the western English Channel in the vicinity of a long-term oceanographic observation site(31). Oligonucleotide probes targeting18S rRNA coupled tofluo-rescent in situ hybridization associated with tyramide signal amplification were used to detect the picoplanktonic taxa(22). Eight new oligonucleotide probes specific for Prasinophyceae were designed and validated on pure cultures.These probes,as well as more general probes targeting the Chlorobionta and the eukaryotes,allowed the study in detail of the dynamics of the dominant taxa along a seasonal time series between July 2000and September2001.MATERIALS AND METHODSCultures.Nineteen unialgal strains of picoeukaryotes belonging to different phylogenetic clades of the Prasinophyceae were selected(Table1).They were grown in Nalgene(Rochester,N.Y.)flasks at20°C in K medium(11).In order to test the new probes designed in this study,cells were harvested during the mid-exponential growth phase.For each culture,4.5ml was harvested andfixed with paraformaldehyde(1%final concentration)for1h.These cultures were filtered on0.2-m-pore-size Anodiscfilters(Whatman International Ltd.,Maid-stone,England).Thefilters were dehydrated in an ethanol series(50,80,and 100%;3min each)and stored atϪ80°C until further hybridization tests were performed.*Corresponding author.Mailing address:Station Biologique,UMR 7127,CNRS et Universite´Pierre et Marie Curie,Place George Teis-sier,BP74,29682Roscoff Cedex,France.Phone:332-98-29-23-70. Fax:332-98-29-23-24.E-mail:vaulot@sb-roscoff.fr.4064Natural samples.Natural samples were collected twice a month between July 2000and September 2001at 0.5-m depth with 5-liter Niskin bottles off Roscoff,France,at the ASTAN station (48°46ЈN,3°57ЈW).The water was pre filtered through a 200-m-pore-size mesh and further processed in the laboratory.Tem-perature,salinity,and concentrations of phosphate,nitrate,and ammonium were measured by standard oceanographic methods.For fluorescent in situ hybridization (FISH),90ml of seawater was pre filtered through 3-m-pore-size Nuclepore filters (Whatman International Ltd.)and fixed with 10ml of 10%paraformaldehyde for 1h.Samples were then filtered onto 0.2-m-pore-size Anodisc filters under a maximum pressure of 200mm Hg and dehydrated in an ethanol series (50,80,and 100%;3min each).The filters were stored at Ϫ80°C.For flow cytometry analyses,1.5ml of pre filtered (3-m pore size)samples was fixed with a mixture of 1%paraformaldehyde and 0.1%glutaraldehyde (final concentrations)and then deep frozen in liquid nitrogen and stored at Ϫ80°C.For high-performance liquid chromatography (HPLC)pigment measure-ments,the Ͻ200-m seawater fraction (1liter)was collected on a GF/F filter (Whatman International Ltd.).The 3-to 200-m fraction was collected on a 3-m-pore-size Nuclepore filter.Finally,the Ͻ3-m fraction was collected on a GF/F filter.The filtrations were conducted under a pressure of 200mm Hg,and all of the filters were immediately deep frozen in liquid nitrogen and stored at Ϫ80°C.Flow cytometry.Total photosynthetic-cell counts were obtained from fixed seawater samples using a FACSsort flow cytometer (Becton Dickinson,San Jose ´,Calif.),as described previously (18).Photosynthetic picoeukaryotes were dis-criminated from cyanobacteria using Cytowin software (available from http://www.sb-roscoff.fr/Phyto/cyto.html).HPLC.Pigment analyses were performed using the method of Zapata et al.(43),with minor modi fications as described by Latasa et al.(14).The contribu-tions of different algal groups to the total chlorophyll a (Chl a )was estimated using CHEMTAX (17).While the contributions of the Mamiellales,Prasinococ-cales,and Pseudoscour fieldiales,which possess prasinoxanthin,could be com-puted,those of the other clades (prasinoxanthinless Prasinophyceae)cannot be distinguished from those of other Chlorophyta.FISH associated with tyramide signal ampli fication.In situ hybridization with horseradish peroxidase-labeled probes,signal ampli fication,and target cell de-tection were performed as described previously by Not et al.(22).The only difference was the use of a more viscous antifading reagent,AF1(Citi fluor Ltd.,London,United Kingdom),instead of AF3in order to preserve the hybridized slides longer (up to 2weeks in the dark at 4°C)without signi ficant loss of fluorescence.Epi fluorescence microscopy and image acquisition.The hybridized cells were observed with an Olympus (Tokyo,Japan)BX 51epi fluorescence microscope equipped with a mercury light source and an 100ϫUVFL (Olympus,Tokyo,Japan)objective.Excitation-emission filters were 360/420for DAPI (4Ј,6Ј-dia-midino-2-phenylindole)and 490/515for fluorescein isothiocyanate.For each sample,10randomly chosen microscopic fields were counted by eye.For probes with broad taxonomic speci ficity (e.g.,CHLO02),Ͼ500cells were counted.Because of the large number of hybridizations and the time required for each analysis,it was not possible to count replicates for each sample.However,for 33samples,three replicates (i.e.,three hybridizations with the same probe on three different filters)were analyzed,and the average error was 15%(range,2to 38%).Design of 18S rRNA oligonucleotide probes.The oligonucleotide probes (Ta-ble 2)were designed with ARB software (16)using a small-subunit rRNA database containing Ͼ30,000complete and partial sequences.In addition to published sequences,our database also contained unpublished partial eukaryote sequences retrieved from three coastal sites.Although the probes could have been designed based only on the public sequences,the additional sequencesTABLE 1.Origins and culture conditions of picoplankton strainsRCC a no.Genus and species Strain Light (E m Ϫ2s Ϫ1)Cell diameter (m)Origin116Ostreococcus tauri OTTH 05951000.8Thau Lagoon143Ostreococcus sp.EUM 13BBL 1000.8Tropical Atlantic Ocean 141Ostreococcus sp.EUM 16BBL 1000.8Tropical Atlantic Ocean 114M.pusilla CCMP b 4901002North Atlantic 299M.pusilla NOUM 171002Equatorial Paci fic 372M.pusilla Naples 1002Gulf of Naples 373M.pusillaSkagerrak 1002Skagerrak 417Mantoniella squamata CCMP 4801003–5North Sea113 B.prasinos CCMP 18981002Mediterranean Sea 369Coccoid CCMP 12051002–6Sargasso Sea287Coccoid NOUM 15100 2.5Equatorial Paci fic Ocean261P.marina TAK 9801404Takapoto Atoll (Paci fic Ocean)370P.provasolii CCMP 12031002–4North Atlantic 135P.provasolii CCMP 11991002–4Gulf of Mexico 251Pycnococcus sp.ROS 94011002English Channel 253Pycnococcus sp.ROS 94041002English Channel 136P.capsulatus CCMP 14071003–6Sargasso Sea 134Prasinococcus MP 11941003–5Gulf of Mexico 137Prasinoderma MP 12201003–8Gulf of Mexicoa Roscoff Culture Collection (http://www.sb-roscoff.fr/Phyto/RCC/).bCCMP (Provasoli-Guillard National Center for Culture of Marine Phytoplankton,West Boothbay Harbor,Maine [/]).TABLE 2.Novel oligonucleotide probes targeting prasinophycean taxaProbe nameSequenceTarget groupPosition of 16S rRNA (Escherichia coli )Closest sequence not targetedTaxonomy No.of mismatchesPRAS015Ј-ACG GTC CCG AAG GGT TGG -3ЈPseudoscour fieldiales clade V 193Tilletia caries2PRAS035Ј-GCC ACC AGT GCA CAC CGG -3ЈPrasinococcales620Friedmannia israeliensis 2PRAS045Ј-CGT AAG CCC GCT TTG AAC -3ЈMamiellales (except the genus Dolichomatix )651Choricystis minor 1PRAS055Ј-GCC AGA ACC ACG TCC TCG -3ЈClade VIIA,RCC 287,CCMP 1205651Pyramimonas olivacea 3PRAS065Ј-AAT CAA GAC GGA GCG CGT -3ЈEnvironmental clade VIIB 651Scutopus ventrolineatus 3MICRO015Ј-AAT GGA ACA CCG CCG GCG -3ЈM.pusilla 211Ostreococcus tauri 1BATHY015Ј-ACT CCA TGT CTC AGC GTT -3Ј B.prasinos 651Uncultivated bacteria3OSTREO015Ј-CCTCCTCACCAGGAAGCU -3ЈOstreococcus647Corynebacterium genitalium3V OL .70,2004PRASINOPHYCEAN PICOEUKARYOTES IN THE ENGLISH CHANNEL4065FIG.1.Phylogenetic tree of the Prasinophyceae obtained by the neighbor-joining method and based on analyses of complete 18S rRNA gene sequences.The speci ficities of the different probes designed in this study are presented.The clades were named according to the system of Guillou et al.(8).The numbers on the branches correspond to bootstrap values (done on 1,000replicates).4066NOT ET AL.A PPL .E NVIRON .M ICROBIOL .allowed us to confirm that the targeted regions were conserved on the corre-sponding sites of coastal representatives of taxa belonging to the clades targeted. When the probes were designed with the ARB Probe Design function,care was taken to maximize the number of mismatches to nontarget sequences and to position these mismatches near the center of the probe.The theoretical speci-ficities of the new probes were checked using the Probe Match function of the ARB software.Oligonucleotide probes with a5Јamino link(C6)were purchased from MWG(Courtaboeuf,France).The probes were then labeled with horse-radish peroxidase(Roche Diagnostic Boehringer,Meylan,France)as described previously(40).The optimal formamide concentration was determined empiri-cally to be40%in the hybridization buffer,and specificity tests of the new probes were performed at this concentration.These probes are available in the rRNA probe database for protists and cyanobacteria(http://www.sb-roscoff.fr/Phyto /Databases/RNA_probes_introduction.php).Because probes with broad taxo-nomic specificity usually do not work equally well for all targeted organisms,the three probes EUK1209R,CHLO01,and NCHLO01were used in combination to estimate all eukaryotes(22).Finally,probe CHLO02,specific for the subregnum Chlorobionta(Chlorophyta and Streptophyta),was applied(30).To our knowl-edge,the division Streptophyta has no representative in the pelagic marine systems.Consequently,in this study,cells labeled by the probe CHLO02were considered to belong to the division Chlorophyta.Tree construction.PAUP*version4.0beta10(33)was used to perform phylogenetic analyses of complete18S rRNA sequences using neighbor-joining methods.Cyanophora paradoxa,Pavlova gyrans,Mesostigma viride,and Chara foetida were included in the analyses as an outgroup,and the tree was rooted with C.paradoxa.Bootstrapping(1,000replicates)allowed the evaluation of tree significance.Trees were drawn using TreeView(Roderic Page,University of Glasgow,Glasgow,United Kingdom).RESULTSDesign and specificity tests of18S ribosomal DNA(rDNA) probes targeting Prasinophyceae.No unique characteristic ex-ists that unites all Prasinophyceae taxa to the exclusion of other Viridiplantae or members of other algal phyla(34).This is also supported by molecular phylogenies that show the paraphyletic structure of the class Prasinophyceae(21).Therefore,it is not possible to identify a single probe targeting all Prasinophyceae species.Based on comparative18S rRNA sequence analyses, we designed eight oligonucleotide probes(Fig.1).The probes PRAS01,PRAS03,PRAS04,PRAS05,and PRAS06are spe-cific,respectively,for the orders Pseudoscourfieldiales(clade V only);Prasinococcales(clade VI);Mamiellales(clade II, with the exception of Dolichomastix);clade VIIA,containing the coccoid strain CCMP1205;and clade VIIB,composed of sequences retrieved from the Pacific Ocean.The probes OSTREO01,BATHY01,and MICRO01are specific for the genus Ostreococcus and the species Bathycoccus prasinos and M.pusilla,respectively(Table2).The specificities of the newly designed probes were evalu-ated by whole-cell hybridization of well-characterized refer-ence strains(Table3).Among the probes targeting Prasino-phyceae orders and clades(PRAS01to-06),only PRAS04 hybridized all,and only,target strains.Since no isolate of clade VIIB has yet been established in culture,positive controls could not be performed for PRAS06,but the probe did not show any nonspecific labeling with the strains tested.PRAS01, PRAS03,and PRAS05hybridized only target strains,but not all of them(Table3).We encountered two different problems. First,PRAS01conferred a goodfluorescence signal when hy-bridized with the two strains RCC261(Pseudoscourfieldia ma-rina)and RCC253(Pycnococcus sp.),but no signal was ob-served with the strain RCC135(Pycnococcus provasolii). Second,RCC369(CCMP1205)and RCC287,targeted by PRAS05,and Prasinoderma strain RCC137,targeted by PRAS03,presented a heterogeneous signal after hybridization: onlyϳ50%of the cells showed a positive signal.The problem encountered with these probes was also observed with the general Chlorophyta probe CHLO02with the same strains. The three genus-and species-specific probes(OSTREO01, BATHY01,and MICRO01)labeled all,and only,the taxa for which they were designed(Table4).Hydrological conditions at ASTAN station.Due to the strong tidal mixing,the coastal waters off Roscoff are perma-TABLE3.Specificity tests of the oligonucleotide probes for Prasinophyceae order level cladesSpecies RCC no.Specificity aPRAS01PRAS03PRAS04PRAS05PRAS06CHLO02M.pusilla114ϪϪ؉ϪϪ؉Ostreococcus tauri116ϪϪ؉ϪϪ؉Mantoniella squamata417ϪϪ؉ϪϪ؉B.prasinos113ϪϪ؉ϪϪ؉Coccoid strain369ϪϪϪ؎Ϫ؎Coccoid strain287ϪϪϪ؎Ϫ؎P.marina261؉ϪϪϪϪ؉P.provasolii135؊ϪϪϪϪ؊Pycnococcus sp.253؉ϪϪϪϪ؉Prasinoderma sp.137Ϫ؎ϪϪϪ؎Prasinococcus sp.134Ϫ؉ϪϪϪ؉P.capsulatus136Ϫ؉ϪϪϪ؉a Boldface indicates the theoretical specificity,whileϩandϪindicate the results of in situ hybridization tests(ϩ,brightfluorescent signal;Ϫ,no detectable fluorescent signal;Ϯ,weakfluorescent signal).TABLE4.Specificities of oligonucleotide probes forMamiellales genera and speciesSpecies RCC no.Specificity aBATHY01OSTREO01MICRO01Ostreococcus tauri RCC116Ϫ؉ϪOstreococcus sp.RCC143Ϫ؉ϪOstreococcus sp.RCC141Ϫ؉ϪB.prasinos RCC113؉ϪϪM.pusilla RCC114ϪϪ؉M.pusilla RCC299ϪϪ؉M.pusilla RCC373ϪϪ؉M.pusilla RCC372ϪϪ؉a See Table3,footnote a,for explanation of symbols.V OL.70,2004PRASINOPHYCEAN PICOEUKARYOTES IN THE ENGLISH CHANNEL4067nently mixed all year round (32).During the sampling period,the temperature varied between 10.2(March 2001)and 16.6°C (August 2001).The nitrate and phosphate concentrations fol-lowed a similar pattern of variation,with minima in summer (0.4and 0.10M,respectively)and maxima in late fall and winter (15.3and 0.66M,respectively)(Fig.2).Photosynthetic pigments.During the period studied,Chl a concentrations in the 0.2-to 200-m fraction (total phyto-plankton)varied according to the classical pattern observed in this area,with minima in winter (0.2g liter Ϫ1)and maxima in summer (2.5g liter Ϫ1),corresponding to the diatom bloom (32).The 2001bloom occurred in late June (Fig.3A).Maxi-mum concentrations of picoplanktonic Chl a occurred later,at the end of July (753ng liter Ϫ1).Large phytoplankton (Ͼ3-m diameter)and picoplankton (Ͻ3-m diameter)contributed approximately equally to the Chl a biomass under nonbloom conditions.During bloom periods (July 2000and July 2001),large phytoplankton dominated the total Chl a biomass.Over-all,picophytoplankton made up 34%of the total Chl a throughout the study period.Within the picoplanktonic Chl a fraction,the contribution of the division Chlorophyta was almost always Ͼ25%(45%,on average)(Fig.3B),except at the end of May,when it plum-meted below 10%,although picoplanktonic Chl a decreased only slightly (Fig.3A).Within the Chlorophyta,the contri-butions of the Mamiellales,Prasinococcales,and some of the Pseudoscour fieldiales (containing prasinoxanthin)to pico-planktonic Chl a ,as estimated by CHEMTAX,were signi ficant all year round and largely dominant (Ͼ80and up to 100%)during spring and summer 2001.Overall composition of the picoeukaryotic community.The abundances of the photosynthetic picoeukaryotes determined by flow cytometry varied between 1ϫ103and 2ϫ104cells ml Ϫ1.Maximum abundances were recorded in May and July 2001(Fig.4A).The abundance of picoeukaryotic cells de-tected by epi fluorescence microscopy after in situ hybridization with a combination of the probes EUK1209R,CHLO01,and NCHLO01varied between 1.3ϫ103and 1.35ϫ104cells ml Ϫ1(Fig.4A).Cells belonging to the division Chlorophyta (tar-geted by CHLO02)dominated the picoeukaryotic community all year long (1ϫ103to 1.5ϫ104cells ml Ϫ1;85%,on average,of the total number of picoeukaryotes)(Fig.4A).Within the Chlorophyta,organisms belonging to the order Mamiellales (Prasinophyceae),targeted by the probe PRAS04,dominated year round (Fig.4B).Within the Mamiellales,M.pu-silla was the dominant species during the sampling period (75%of the cells detected by PRAS04,on average)(Fig.4C)and accounted for 45%of picoplanktonic eukaryotes.The sec-ond most abundant species detected was B.prasinos ,which accounted for 12%(range,1to 67%)of the Mamiellales and 8%of the picoplanktonic eukaryotes (Fig.4C).Cells belonging to the genus Ostreococcus were less abundant,with an average of 3%(range,0to 19%)of the Mamiellales and 1.4%of the eukaryotes (Fig.4C).The second most abundant clade within the division Chlo-rophyta was the order Prasinococcales,targeted by the probe PRAS03and accounting for an average of 3.4%(range,0to 34%)of the Chlorophyta cells (Fig.4B).Cells belonging to the other clades were detected at very low abundances all year long.Probes PRAS01,PRAS05,and PRAS06targeted an av-erage of 0.6,0.4,and 0.6%,respectively,of cells belonging to the division Chlorophyta (Fig.4B).Only 16%,on average,of the cells detected by the probe speci fic for the division Chlo-rophyta were not targeted by the Prasinophyceae probes used in this study.Seasonal variation of picoeukaryotic community.The pico-eukaryotic community exhibited a marked seasonal cycle,with minima of abundance in early winter and maxima (ϳ10times the winter abundances)in midsummer.The patterns of abun-dance variation during the sampling period were similar for total picoplanktonic eukaryotes,photosynthetic picoeukary-otes,Chlorophyta,and Mamiellales and for the speciesM.FIG.2.Variations in temperature and phosphate and nitrate concentrations at the ASTAN station between July 2000and September 2001.4068NOT ET AL.A PPL .E NVIRON .M ICROBIOL .pusilla .On a smaller temporal scale,three major peaks of abundance were observed in 2001,in late spring and summer (mid-May,mid-June,and the end of July)(Fig.4A).For the Mamiellales and for M.pusilla ,an additional peak was ob-served in late August (Fig.4C).The other clades and genera detected in this study showed more sporadic proliferations throughout the year.The species B.prasinos was detected all year long but became more abundant during spring (1.8ϫ103cells ml Ϫ1in March 2001)(Fig.4C).The cells targeted by the probe PRAS03(order Prasinococcales)reached signi ficant abundance in early fall (1.2ϫ103cells ml Ϫ1)and late spring (600cells ml Ϫ1).In fall 2000,they accounted for up to 34%of the cells targeted by the CHLO02probe (Fig.4B).Spectacular decreases in the cell abundances of all the taxa detected by our probes and in flow cytometry counts were observed on May 30,2001.DISCUSSIONNew probes targeting Prasinophyceae.Phylogenetic analy-ses of 18S rDNAs recovered directly from samples collected in different oceanic regions have revealed an unsuspected diver-sity within eukaryotic picoplankton (20).In most gene libraries analyzed,including libraries constructed from samples col-lected in the western English Channel (26),prasinophycean sequences were well represented (42).The set of probes pre-sented in this study covers most known clades (Fig.1)and,compared to the few previously published probes for prasino-phycean taxa,have improved speci ficities.Indeed,PRAS04has no mismatch with any Mamiellales sequence known to date,in contrast to PRAS02(2),which has one mismatch with B.pra-sinos .The probes Micro/Manto1and Micro/Manto2,designed in 1996(12)for Micromonas and Mantoniella ,presentmatchesFIG.3.(A)Variations in Chl a biomass as measured by HPLC (total Ͻ200-m and fraction Ͻ3-m)at the ASTAN station.(B)Contributions of the division Chlorophyta and of the orders Mamiellales,Prasinococcales,and Pseudoscour fieldiales to the picoplankton fraction (Ͻ3-m diameter)according to the CHEMTAX algorithm applied to HPLC data.V OL .70,2004PRASINOPHYCEAN PICOEUKARYOTES IN THE ENGLISH CHANNEL 4069with the genus Ostreococcus and mismatches with some strains of Micromonas .They are therefore not speci fic for their initial targets (data not shown).When tested against pure cultures,probes PRAS04,MICRO01,OSTREO01,and BATHY01showed perfect spec-i ficity for their target strains and delivered a bright fluorescent signal (Tables 3and 4).In contrast,we encountered hybrid-ization problems with some strain-probe combinations,i.e.,either cells showed no fluorescent signal and a strong red fluorescence under blue excitation or the fluorescent signal was heterogeneous.Given that the corresponding sites on the 18S rRNA sequences of the strains tested showed 100%homology with the probes,the most likely hypothesis is that the probes did not penetrate into the cells.In fact,the redchlorophyllFIG.4.Abundances (number of cells ml Ϫ1)of photosynthetic picoplankton off Roscoff (western English Channel)between July 2000and September 2001.The average percentages over the time series of the different groups are represented in pie charts.(A)Picoeukaryotic photo-synthetic cells detected by flow cytometry counts (Flow Cytometry);picoeukaryotic cells targeted by the mix of the general probes EUK1209R,CHLO01,and NCHLO01(EUK1209R ϩCHLO01ϩNCHLO01);and cells belonging to the division Chlorophyta detected by the probe CHLO02(CHLO02).(B)Cells targeted by the probe CHLO02and cells detected by the probes speci fic for the clades PRAS01,PRAS03,PRAS04,PRAS05,and PRAS06.(C)Cells targeted by the probe speci fic for Mamiellales (PRAS04)and cells detected by the probes (MICRO01,BATHY01,and OSTREO01)speci fic for the species and genera.4070NOT ET AL.A PPL .E NVIRON .M ICROBIOL .fluorescence remaining in these strains after ethanol treatmentsuggests that their cell walls are probably very thick and resis-tant,a fact previously established for P.provasolii and Prasi-noderma sp.(9),and therefore prevent probe penetration. Analysis of the picoeukaryotic community.The seasonal variation in microphytoplankton diversity and abundance intemperate sea waters has been well described.Off Roscoff,themicrophytoplankton bloom occurs in late spring(with cell den-sities up to3ϫ104cells literϪ1)and is characterized by thesuccession of a very limited number of diatom species(such asGuinardia delicatula)that occurs at low concentration outside this period(S.Ristori,unpublished data;32).The presentstudy indicates that the structure and dynamics of the pico-planktonic community are totally different from the classicalscheme observed for microphytoplankton.First,picophyto-plankton abundance remains high all year round(1ϫ103to2ϫ104cells mlϪ1).Hence,this size class contributes an average of50%of the total Chl a biomass under nonbloom conditions.Second,both FISH and chemotaxonomic analyses show thatgreen algae,and more precisely Prasinophyceae,dominate thecommunity(70%of the cells and35%of the total picoplank-tonic Chl a).Chlorophytes and Prasinophyceae have beenpreviously identified by chemotaxonomic analyses(i.e.,thepresence of Chl b)as major components of the nano-orpicoplankton size fractions in the English Channel(3),as wellas in other coastal systems,such as the Faroe-Shetland Chan-nel(20%of total Chl a)and the Galician coast(60%of totalChl a in winter)(24,25).Most interestingly,FISH data allowed us to establish unam-biguously that a single species,M.pusilla,is dominant all yearround and shapes the dynamics of the whole picophytoplank-ton ing published values of Chl a content percell for Micromonas(0.025pg cellϪ1)(5)and cell abundancesobtained by FISH,the contribution of Micromonas to the totalChl a can be estimated to range between2and50%(22%,onaverage)of the picoplanktonic Chl a.M.pusilla has alreadybeen detected in different polar and temperate marine systems,including coastal and open-ocean regions,such as the Norwe-gian coast(38),Bay of Biscay(1),Arctic Sea(36),Mediterra-nean Sea(44),and Skagerrak(13),with abundances rangingbetween1ϫ102and9ϫ103cells mlϪ1.The species has also been found to be present year round in the Skagerrak and North Sea(13)and to be abundant during winter-spring in the Mediterranean Sea(39)or spring and summer in the North Atlantic and North Sea(13).The contributions of other Mamiellales genera to the pico-planktonic community were less important.The contribution of the nonmotile picoplanktonic species B.prasinos,described from the Mediterranean Sea(6)and reported from the Atlan-tic and Pacific Oceans(28)and Norwegian coastal waters (6),is much more sporadic:for example,in spring2001,it accounted for30%of picoeukaryote cells targeted.The genus Ostreococcus was detected,but always at low abundance,off Roscoff.Prior to this study,it had been found in abundance only in a somewhat atypical ecosystem,the Thau lagoon,a shallow ecosystem used intensively for oyster culture(4).The second most abundant group within the Prasinophyceae is the order Prasinococcales,which contains species such as Prasino-coccus capsulatus,which has been observed and is suspected to be significant in the western Atlantic and the western Pacific (27).The persistence of high abundances of picophytoplankton year round could be explained by the fact that picophytoplank-ton is controlled by small predators with short generation times (29)that prevent any sudden cell proliferation,even when conditions are optimal.This contrasts with what happens for diatoms controlled by copepods with long generation times and complex life cycles,which are able to reduce biomass only after a delay,allowing blooms to develop.The strong domi-nance of M.pusilla over other species all year in the system studied is very similar to what is observed for picophytoplank-tonic prokaryotes,among which single genera,such as Prochlo-rococcus and Synechococcus,dominate certain types of ecosys-tems(23).Micromonas may have a broad ability to respond to light and nutrient variations and thus,because of this large environmental spectrum,may be little affected by changes in the environment.However,during the seasonal series studied, a sharp decrease in abundance of picoeukaryotes(and M.pu-silla)was recorded at the end of May.This phenomenon was associated with a major peak in the ammonium concentration (data not shown).One explanation for this decrease could be that Micromonas was infected by viruses.Previous studies have suggested that viruses infecting M.pusilla have a profound impact on populations of the species in natural systems(44). Most surprising is the ability of Micromonas populations to recover rapidly to their initial levels only15days after the decrease.This upturn could be due to the proliferation of a different Micromonas genotype,resistant to the virus respon-sible for the decay.Indeed,a recent study based on the analysis of18S rDNA gene sequences demonstrated the presence of three clades within the species M.pusilla(8).Our results suggest that the taxonomic structure of the pi-coplanktonic communities is different from the structure of the microphytoplanktonic community,with one species(M.pu-silla)being dominant all year round off Roscoff.Hence,if the diatoms,dinoflagellates,and coccolithophorids(i.e.,Chl c-con-taining lineages)dominate the large eukaryotic phototrophic assemblages in the contemporary oceans(7),a few species of the green(Chl b-containing)lineage may dominate picoplank-ton and shape the marine microbial food webs.The next step will be to extend these observations to other coastal and oce-anic areas in order to determine how widespread M.pusilla dominance is and to understand the factors that regulate the diversity and abundance of picoeukaryotes.ACKNOWLEDGMENTSWe thank Isabelle Biegala and Carlos Pedro´s-Alio´for constructive discussions,Laure Guillou and Erwan Corre for help with phyloge-netic analyses,the MYSIS crew for efficient assistance with sampling, and Florence Le Gall for help with phytoplankton cultures.This work was funded by the following programs:PICODIV,sup-ported by the European Union(EVK3-CT-1999-00021);PICMANCH, supported by the Re´gion Bretagne;and BIOSOPE,supported by PROOF(CNRS).F.N.was supported by a doctoral fellowship from the French Research Ministry.REFERENCES1.Ansotegui,A.,A.Sarobe,J.M.Trigueros,I.Urrutxurtu,and E.Orive.2003.Size distribution of algal pigments and phytoplankton assemblages in a coastal-estuarine environment:contribution of small eukaryotic algae.J.Plankton Res.25:341–355.V OL.70,2004PRASINOPHYCEAN PICOEUKARYOTES IN THE ENGLISH CHANNEL4071。
关于2024年315事件英语作文

关于2024年315事件英语作文The 2024 315 Incident was a pivotal moment that shook the world and forever changed the course of history. On March 15th, 2024, a series of coordinated cyberattacks targeted the global financial system, causing widespread chaos and disruption that reverberated across every corner of the planet.The attacks began with a sophisticated malware that infiltrated the servers of major banks, stock exchanges, and financial institutions around the world. Within hours, trillions of dollars were wiped off the global markets, sending shockwaves through the international economy. Panicked investors scrambled to salvage what they could, leading to a massive sell-off that only exacerbated the crisis.As the full scale of the disaster became clear, governments and central banks struggled to respond. Attempts to stabilize the markets and restore confidence were met with limited success, as the malware continued to wreak havoc on the financial infrastructure. Transactions were frozen, accounts were drained, and the very foundations of the global financial system seemed to be crumbling.The impact was felt far beyond the world of finance. Businesses large and small were crippled by the economic turmoil, leading to widespread layoffs, bankruptcies, and a sharp decline in consumer spending. Governments were forced to divert resources away from critical public services to address the crisis, further straining already stretched budgets.The human toll was equally devastating. Millions of people around the world saw their life savings vanish overnight, plunging them into financial ruin and despair. The stress and anxiety caused by the crisis took a heavy toll on mental health, with reports of a surge in depression, anxiety, and even suicides.As the world struggled to come to terms with the magnitude of the disaster, the search for the perpetrators began in earnest. Suspicion quickly fell on a group of highly sophisticated hackers with ties to a foreign state, but the investigation proved to be a complex and arduous process.Tensions escalated as nations pointed fingers and accused one another of orchestrating the attacks. Diplomatic relations were strained, and the threat of retaliation loomed large. The world seemed to be on the brink of a global conflict, with the 315 Incident serving as a catalyst for a new era of geopolitical tensions anduncertainty.In the aftermath of the crisis, the world was forced to confront the vulnerabilities of the global financial system and the urgent need for comprehensive cybersecurity measures. Governments and private sector organizations alike scrambled to implement new safeguards and protocols to prevent such a catastrophic event from happening again.The 315 Incident also sparked a renewed debate about the role of technology in modern society and the ethical implications of its use. Questions were raised about the responsibility of tech companies, the need for greater regulation, and the delicate balance between innovation and security.As the world slowly recovered from the 315 Incident, it became clear that the event had forever changed the landscape of global finance and geopolitics. The scars left by the crisis would take years, if not decades, to heal, and the lessons learned would shape the course of history for generations to come.。
The rotation curve and mass-distribution in highly flattened galaxies

˜ is defined in the following way: and the function Φ 1 ˜ ρ, z Φ(˜ ˜) = 2 2π
π ∞
dθ
0 R
ln[(s − ρ ˜ cos θ)2 + z ˜2 ] α(s) ds ,
(3)
α(s) being a real function which has the meaning of the density of sources distribution. Due to the specific analytical properties of the logarithmic function which turn out to be most advantageous for fulfilling the Laplace equation in the region exterior to disk’s sources, hereafter we shall be able to relate the function α(s) to the surface density σ (ρ) of the disk and to the distribution of the angular velocity ω (ρ) via simple integral formulas. ˜ by The potential Φ(ρ, z ) which, as can be easily seen from (1), is connected with Φ Thomson’s transform, satisfies Laplace’s equation everywhere except on the disk. On the circle z = 0, ρ ≤ R the normal derivative ∂ Φ/∂z experiences a jump. To show this, let us tend z to zero. Then, by virtue of (2), we have ˜ ∂Φ ρ ˜3 ∂Φ = 2 lim , z →+0 ∂z ˜→+0 ∂ z R z ˜ lim while from (3) follows that ˜ ∂Φ 1 lim = lim z ˜→+0 ∂ z ˜→+0 2π 2 ˜ z
Revealing Lives from the Pharaohs Era

Revealing Lives from the Pharaohs Era The Pharaohs era in ancient Egypt is a fascinating period of history that continues to captivate our imagination. Through the study of archaeological findings, we are able to catch a glimpse into the lives of these powerful rulers and the society they governed. The discovery of tombs, temples, and artifacts has provided valuable insights into the daily lives, beliefs, and customs of the Pharaohs and their subjects. One of the most iconic aspects of the Pharaohs era is the elaborate burial practices and rituals that were performed to ensure a smooth transition into the afterlife. The construction of monumental tombs, such as the pyramids of Giza, and the intricate burial chambers filled with treasures and offerings, reflect the belief in the afterlife and the importance of preserving the Pharaoh's legacy for eternity. The meticulous care taken in the mummification process further demonstrates the reverence and awe in which the Pharaohs were held. Beyond the grandeur of their tombs, the Pharaohs were also responsible for governing and maintaining order in ancient Egypt. They held absolute power and were considered living gods by their subjects. The Pharaohs were not only political leaders but also religious figures, responsible for maintaining the balance of Ma'at, the cosmic order of the universe. They oversaw the construction of temples, monuments, and irrigation systems, ensuring the prosperity and well-being of their people. Despite their divine status, the Pharaohs were also human beings who faced challenges and struggles like any other ruler. Succession disputes, foreign invasions, and internal unrest were common occurrences during the Pharaohs era. The reign of certain Pharaohs, such as Akhenaten, who attempted to introduce monotheism to Egypt, or Hatshepsut, one of the few female Pharaohs, were marked by controversy and intrigue. These personal and political struggles humanize the Pharaohs, reminding us that they were not infallible beings but individuals with their own ambitions and flaws. The lives of the Pharaohs were intricately intertwined with those of their subjects, who played a crucial role in supporting and upholding the authority of the Pharaoh. The social hierarchy of ancient Egypt was rigidly structured, with the Pharaoh at the top, followed by nobles, priests, scribes, artisans, and farmers. Each segment of society had its own responsibilities and duties, contributing to the overallfunctioning of the kingdom. The Pharaohs relied on the loyalty and obedience of their subjects to maintain their power and ensure the stability of the kingdom. In conclusion, the Pharaohs era offers a rich tapestry of history, culture, and mythology that continues to intrigue and inspire us today. By delving into the lives of these ancient rulers, we gain a deeper understanding of the complexities of ancient Egyptian society and the enduring legacy of the Pharaohs. Through the study of archaeological evidence and historical records, we are able to piece together the puzzle of the Pharaohs era, revealing the triumphs and tribulations of these iconic figures who shaped the course of history.。
我的偶像c罗英语作文

Cristiano Ronaldo, a name that resonates with passion, dedication, and excellence in the world of football. Born on February 5, 1985, in Funchal, Madeira, Portugal, Ronaldo has become a living legend, not just in his home country, but across the globe. As my idol, he has inspired me in more ways than one can imagine.From a young age, Ronaldo displayed an unmatched zeal for football. His journey from a humble beginning to becoming a global icon is nothing short of extraordinary. He started his professional career with Sporting Lisbon, where his talent was quickly recognized, leading to a move to Manchester United. Under the guidance of Sir Alex Ferguson, Ronaldo honed his skills and became a key player, winning numerous titles, including three Premier Leagues and the UEFA Champions League.However, it was his move to Real Madrid in 2009 that catapulted him to superstardom. With his incredible speed, agility, and goalscoring prowess, Ronaldo broke record after record, becoming Real Madrids alltime top scorer and helping the team secure multiple UEFA Champions League titles. His rivalry with Barcelonas Lionel Messi was intense, pushing both players to greater heights and captivating football fans worldwide.Ronaldos dedication to his craft is aweinspiring. His rigorous training regimen, commitment to fitness, and relentless pursuit of perfection have set a new standard for athletes. He is known to arrive at training hours before his teammates, often staying behind to practice free kicks and other skills. This level of dedication is a testament to his character and work ethic.Off the field, Ronaldo is a philanthropist, using his fame and wealth to give back to society. He has donated millions to various charitable causes, including disaster relief efforts and childrens hospitals. His generosity and kindness have touched the lives of many and serve as a reminder that true greatness extends beyond the football pitch.In 2018, Ronaldo made a bold move to Juventus, an Italian club with a rich history. His arrival in Turin was met with great anticipation, and he has continued to dazzle fans with his performances. Despite facing new challenges and adapting to a different style of play, Ronaldo remains a force to be reckoned with, consistently delivering goals and leadership to his team.As my idol, Cristiano Ronaldo has taught me the importance of hard work, perseverance, and humility. His journey from a young boy in Madeira to a global sports icon is a story of determination and resilience. He has shown that with passion and dedication, one can overcome obstacles and achieve greatness.In conclusion, Cristiano Ronaldo is more than just a football player he is a symbol of inspiration, a beacon of hope for those who dare to dream big. His accomplishments on and off the field have made him a role model for millions, and I am proud to call him my idol. As I continue on my own journey, I strive to embody the qualities that make Ronaldo such an extraordinary individual: unwavering commitment, relentless pursuit of excellence, and a genuine desire to make a positive impact on the world.。
英语报纸2024高一31期作文

英语报纸2024高一31期作文2024高一31期英语报纸The Rising Stars: How High School Students in 2024 Are Making a DifferenceIn today's issue, we are proud to showcase the amazing achievements of high school students in 2024 who are making a difference in their communities and beyond. From advocating for climate change action to creating impactful social campaigns, these young individuals are proving that age is just a number when it comes to making a positive impact on the world.One shining example is Emily, a 16-year-old student from Green Valley High School who has been leading a campaign to raise awareness about the dangers of plastic pollution in the oceans. With the support of her classmates, Emily has organized beach clean-up events, lobbied local businesses to reduce their plastic usage, and even started a petition to ban single-use plastics in her city. Her dedication and passion have inspired her peers to take action and make a difference in their own communities.Another inspiring story comes from Alex, a 17-year-old student from Oakwood High School who has been using hisplatform as a social media influencer to raise awareness about mental health issues among teenagers. Through his engaging videos and posts, Alex has started important conversations about the stigma surrounding mental illness and the importance of seeking help. He has also organized fundraisers for mental health organizations and connected with other students who are struggling with their mental health, offering them support and encouragement.These are just a few examples of the incredible work that high school students are doing in 2024. From volunteering at local charities to organizing fundraisers for important causes, these young individuals are proving that they are not just the leaders of tomorrow, but the leaders of today.As we celebrate the achievements of these rising stars, we are reminded of the power of youth and the potential for positive change that lies within each and every one of us. Let's continue to support and encourage the young leaders of today as they work towards creating a brighter future for all.Stay tuned for more inspiring stories from high school students in our next issue. Until then, keep shining bright and making a difference in your own unique way. The world is waiting for your light to shine!。
Gamma函数的相关函数及其q化的完全单调性

Gamma函数的相关函数及其q化的完全单调性李晶晶;孙梅【摘要】A investigation to derive the complete monotonicities of functions involving the Gam-ma function and their q-analogues is presented.First of all,using the integral representations and the monotonicity theory,it is proved that the complete monotonicities of functions Fα(x)=ψ'(x)+α/x-1/(x+1),fα(x)=lnx-ψ(x)-α/x and f(x)=lnx-ψ(x)+1/x2 on(0,∞);Second,based on the properties of q-analogues,the complete monotonicities of q-analogues about these three functions are respectively discussed by the series expansion of functions,the monotonicity theory and induction,and two inequalities are obtained because of the property.%为研究Gamma函数的相关函数及其q化的完全单调性,首先利用函数的积分表达式以及单调性理论证明函数Fα(x)=ψ'(x)+α/x-1/(x+1),fα(x)= lnx-ψ(x)-α/x以及f(x)=lnx-ψ(x)+1/x2在(0,∞)上的完全单调性;其次基于q化的性质,利用函数的级数展开、单调性理论以及归纳法讨论上述相应函数的q化的完全单调性,并利用该性质得到2个不等式。
西方神话英语作文

Western mythology is a rich tapestry of stories,characters,and themes that have been passed down through generations.It encompasses a variety of cultures,including Greek,Roman,Norse,and Celtic,each with its own unique set of myths and legends. Here,we will explore some of the key elements of Western mythology and how they have influenced literature,art,and culture.1.Greek Mythology:Greek mythology is perhaps the most wellknown of the Western mythologies.It features a pantheon of gods and goddesses,each with their own distinct personalities and powers.Zeus,the king of the gods,is known for his thunderbolts,while Aphrodite,the goddess of love,is celebrated for her beauty.The stories of Greek mythology often revolve around the struggles and triumphs of both gods and mortals, such as the labors of Hercules or the tragic tale of Orpheus and Eurydice.2.Roman Mythology:Roman mythology was heavily influenced by Greek myths,with many of the same gods and goddesses,but with different names.For example,Zeus is known as Jupiter in Roman mythology.Romans also added their own unique myths,such as the story of Romulus and Remus,the twin brothers who were said to have founded Rome after being raised by a shewolf.3.Norse Mythology:Norse mythology,originating from the Scandinavian region,is known for its epic tales of gods,heroes,and the struggle between good and evil.The Norse gods,including Odin,Thor,and Loki,are central figures in these stories.The concept of Ragnarok,the end of the world and the final battle between the gods and the giants,is a significant theme in Norse mythology.4.Celtic Mythology:Celtic mythology is characterized by its rich oral tradition and stories of heroes,magical creatures,and the Otherworld.The Celts believed in a world intertwined with the natural and supernatural.Stories often involve quests,magical objects,and the Tuatha DéDanann,a race of supernatural beings.5.Heroes and Heroines:A common theme in Western mythology is the heros journey. Heroes like Perseus,Theseus,and Beowulf are celebrated for their bravery and strength. They often face seemingly insurmountable challenges,such as battling monsters or overcoming personal flaws,which they must overcome to achieve greatness.6.Creation Myths:Many Western mythologies include stories about the creation of the world.In Greek mythology,for example,the world was created from Chaos,and the first gods emerged from this primordial state.Similarly,in Norse mythology,the world was formed from the body of the giant Ymir.7.Moral Lessons:Western myths often contain moral lessons or cautionary tales.For instance,the story of Icarus,who flew too close to the sun and fell to his death,serves as a warning against hubris and overconfidence.8.Cultural Impact:The stories of Western mythology have had a profound impact on Western culture.They have influenced literature,with many authors drawing on mythological themes and characters.They have also inspired art,music,and even modernday movies and television shows.9.Modern Interpretations:Today,Western mythology continues to be reinterpreted and adapted in various forms.From the Percy Jackson series to the Marvel Cinematic Universes Thor,these ancient stories are given new life and continue to captivate audiences.10.Study and Research:The study of Western mythology is not just limited to literature and history it also extends to fields like psychology,where scholars explore the psychological significance of myths and their characters,such as the work of Carl Jung on archetypes.In conclusion,Western mythology is a treasure trove of stories that have shaped and continue to influence our understanding of the world,human nature,and the divine. Whether its through the epic tales of gods and heroes or the moral lessons embedded within these narratives,the myths of the West continue to resonate with people across cultures and time periods.。
- 1、下载文档前请自行甄别文档内容的完整性,平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益,请联系客服反馈,我们会尽快为您处理(人工客服工作时间:9:00-18:30)。
a r X i v :h e p -p h /0011085v 1 7 N o v 20001Logarithmically rising hh and γh cross-sections :some features of model versus experimental data.E.MartynovaaBogoliubov Institute for Theoretical Physics,National Academy of Sciences of Ukraine,03143,Kiev-143,Metrologicheskaja 14b,Ukraine e-mail:martynov@bitp.kiev.uaA Regge model of the Pomeron with intercept equal to one,leading to rising cross-sections is considered.Analysis of the experimental data on hadron and photon induced interactions is performed within the model.It is shown that the available pure hadronic data as well as data on DIS are compatible with a high energy logarithmic behaviour of cross-sections and do not require a Pomeron intercept above one.The very important rˆo le of the preasymptotic contributions is especially emphasized.In this talk I would like to discuss the impor-tance of preasymptotic terms when the available data are analyzed and described in the frame-work of Regge approach.These terms strongly influence the conclusions about properties of the asymptotic term,the Pomeron.I shall explicit as an example the soft Pomeron model,not vio-lating the Froissart-Martin bound,in which the contribution of secondary Reggeons,in particu-lar f Reggeon,can not be neglected,even at the highest accelerator energies.The model describes the hadron elastic scattering as well as the proton structure function.The elastic scattering amplitude in Regge ap-proach is given by the sum of Pomeron and sec-ondary ReggeonsA (s,t )=P (s,t )+R (s,t )≡P (s,t )+f (s,t )+[±ω±ρ±a 2,...]where the signs of terms in square brackets de-pend on the process under consideration.I concentrate here firstly on two aspects :the exchange degeneracy of various Reggeons and the phenomenological separation between the Pomeron and the f Reggeon.•Exchange degeneracy.The hypothesis is based on the fact that f,ω,ρ,a 2trajectories seem to coincide on a Chew-Frautschi plot.The linear parameterizationαe −d (m 2)=αe −d (0)+α′e −d m2of the single exchange-degenerate trajectory,fit-ted to the data,gives [1]αe −d (0)=0.449,α′e −d =0.901GeV−2and an extremely high χ2,namely χ2/dof ≈118.Fitting in isolation each of the trajectories,we obtain the result shown in Fig.1.Numerically we haveαf (0)=0.697±0.041,α′f =(0.801±0.002)GeV−2,χ2/dof =6.01,αω(0)=0.436,α′ω=0.923GeV−2,(not fitted),αρ(0)=0.478±0.001,α′ρ=(0.880±0.002)GeV−2,χ2/dof =3.31,αa 2(0)=0.512±0.041,α′a 2=(0.857±0.023)GeV −2,χ2/dof =0.42.The bad χ2for the f trajectory is due to its evi-dent nonlinearity (see below).Thus the first conclusion is the following :the available data on mesons lying on the f ,ω,ρand a 2Regge trajectories contradict the exchange degeneracy assumption.It should be noted that,in accordance with the conclusion derived in [1,2]1,the hypothesis of exchange degenerate trajectories is not supported also by the forward scattering data.2trajectories taken separately assuming linearity (the figure below is an enlargement for small masses).•f trajectory.The nonlinearity of the f trajectory is illustrated unambiguously in Fig.2,where the above mentioned linear fit is shown to-gether with the parabola passing exactly through the three known resonances.Fig.2Real part of the f trajectory versus thesquared mass of the resonance,m 2.Solid line is the parabola passed through the known resonances.Dashed straight line is the result of a linear fit.Certainly,trajectories must rise slower than first power of t at t →∞.A more realistic f tra-jectory chosen for example in the formαf (t )=αf (0)+β1(4m 2π−t )+β2(√t 1−t )leads to 0.77<αf (0)<0.87.Thus,we obtain a very interesting and impor-tant phenomenological consequence of the non-linearity :a higher intercept of the f ing to the next conclusion :the intercept of the f trajectory is ≥0.7,but most probably,the lower bound is larger than this value.•Separation of Pomeron and f Reggeon.Generally,there is an evident correlation between the intercept of the f Reggeon and the model for the Pomeron.This is due to the fact that in all known processes Pomeron and f Reggeon con-tribute additively.As a rule,a higher f intercept is associated with a slower growth with energy due to the Pomeron contribution [3].In Fig.3we illustrate this observation and show how αf (0)is correlated with a power of ℓns in the behaviour of the total cross-section,if the forward pp (¯p p )scat-tering amplitudes are parametrized in the formA (hh )(s,0)=i C 1+C 2ℓn γ(−is/s 0)+f (s,0)±ω(s,0),with the standard form of secondary Reggeons contribution.Fig.3Intercept of the f trajectory correlated withthe power γof ℓns in σtot (s ).Intervals for intercept of some trajectory parameterizations are shown.Taking into account the restriction αf (0)>0.7,revealed from spectroscopy data,one can con-clude that γ<2or that total cross-section rises slower than ℓn 2s .3On the other hand it is known [1–5]from a comparison of various Pomeron models that the ”best”description (or χ2)is obtained if the Pomeron contribution to the total cross-section has the form σP =C 1+C 2ℓn (s/s 0).The dif-ference in χ2is quite small (∼a few percents)if √s min =9GeV.An additional advan-tage of the logarithmic model is its stability (ofthe parameters as well as the χ2)when√s 0)∆is generalized for two com-ponents i [˜C1+˜C 2(−is/s 0)∆](provided the Regge trajectories are non exchange degenerate)then a fit to the total cross-sections leads to a very small value of ∆(≈0.001).Hence,one can approximate ˜C1+˜C 2(−is/s 0)∆≈C 1+C 2ℓn (−is/s 0),C 1=˜C 1+˜C 2,C 2=∆˜C 2with all parameters (C 1,C 2as well as couplingsand intercepts of the secondary Reggeons)co-inciding with those obtained in the logarithmic Pomeron model.This observation is valid not only for pp and ¯p p cross-sections but also for all hadronic,γp and γγcross-sections.It is necessary to emphasize that in this model the preasymptotic f term gives quite a large part of the total cross-section even at the Tevatron en-ergy,σf (√s ≥5GeV for t =0[5]and√Q 2(1−x )(1+4m 2p x 2/Q 2)F 2(x,Q 2),where σγ∗pL =0is assumed.The forward scatter-ing at W 2=Q 2(1/x −1)+m 2p being far from the threshold W th =m p is dominated by the4Pomeron and the f Reggeon2A (W 2,Q 2;t =0)=P +f,P =P 1+P 2,P 1=iG 1(Q 2)ln(−iW 2m 2p)αf (0)−1(1−x )B f .It evidently follows from the experimental datathat Q 2σγ∗p (W,Q 2)decreases with Q 2,at least at high Q 2.We chooseG i (Q 2)=g i2Weignored an a 2Reggeon considering the f term as an ”effective”one at W ≥3GeV.3An attempt to extract such a local slope as a function of x and Q 2is given in [8].intervals of x .The investigated Dipole Pomeron has an intercept exactly equal to one.Neverthe-less,due to interferences Pomeron-f Reggeon,the model is able to describe well the data on aver-aged x -slope as it is shown in Fig.4.Fig.4The effective intercept calculated in theDipole Pomeron model [7](triangles and solid line)compared with the experimental data.We have shown that a careful account of the preasymptotic contributions allows to describe the available data within the model that does not violate the asymptotic bounds on the cross-sections.REFERENCES1.P.Desgrolard,M.Giffon,E.Martynov,E.Pre-dazzi,hep-ph/00062442.J.R.Cudell et al.,Phys.Rev.D 61(2000)0340193.P.Desgrolard,M.Giffon,E.Martynov,NuovoCim.,110A (1997)5374.P.Desgrolard,M.Giffon,A.Lengyel,E.S.Mar-tynov,Nuovo Cim.107A (1994)6375.P.Desgrolard,M.Giffon,E.Martynov,E.Pre-dazzi,Eur.Phys.J.C 9(1999)6236.P.Desgrolard,M.Giffon, E.Martynov,hep-ph/0004150,to be published in Eur.Phys.J.7.P.Desgrolard,A.Lengyel, E.Martynov,Eur.Phys.J.C 7(1999)6558.P.Desgrolard,A.Lengyel,E.Martynov,theseProceedings.。