=..

Thomas Fried[, Natatiya Rybalka and Anastasiia Kryvenda

2 Phylogeny and systematics of microalgae:

An overview
2,1 lntroduction
Al.!,ae is a Lcrrl oI convenience thirt refers to a collectiolt 0f hlghly cliverse organ-
isms that citrry out photosyrthcsis aud/or possess plastids (l(eeling 2004). l\4any
authors cven include llre prol(arvotic c]/anoll.rctcria Nith algae becar-lsc thcy cxhibit
a lifestyle rather similar kJ their euharl'otic counterparts ancl oftcn share the same
habitat \,vith them. I{icroal.e,ac suitallle for biotechnological cxploitation sltoul.l be
eits_v to gro\\' ir.i large (lLrantities itnd robust in all stages of prrx:essing. Such algac
may be found in almost all linc.ages of cvanobacteria and erLkarl otcs. Examples of
microailgiae (inclucling cyanobacteria) currc[tly used in biotechnological applica
tions or with high poleniial for biotechnological c,.iploitatiol.l are shown in Figures
2.1 and 2.1. i\rlost suit('d arc uniccllular algae $'ith rell rvalls, i.e. of coccoid orgitni
zation, without arttacl-rmcnl stmctures to prevent biofoulitrg r,vhile pror:essing, ancl
thcy sl.rorlkl not form resting stagcs (c.-e,. through sexual rcproduction) ul.tder
adverse cultLrre conclitions. Desl)ile th-.ir apparcnt morphological sintlllicit!, sLLch
algac ollen belong to (litfercnt unrclatccl phylogcneljc lineages aDd, thctcfore, ntity
also cxhibit cliffcrcnt phvsiologi.al anil lliochcntical properties. For examplc. coc
coid eul(aryotic nticroalgac, rvhich irr.-" from very r'listantly rclatcd cvolutionary line
ilges, lllay look sup-.rlir:ially rather similar at low nagnification (see !ig. 2.2c.d).
hr light of the extrcmc divcrsity at.]d exciting ."\/olutionary historlr oI lnic(ralgae
(Fi!a. 2.1), it is surprising that so far only a I'ery ni1lot [ra.{ion ol the a]gal .livcrsitlr
Iias been explorecl for biotcchnologirai applit:alions (1ah. 2.1). Currcntiy, lhere are
probably nq nore than about l0 spccics and gcncra front no nore than 11 taxo-
nomi. r'lasscs of autotrophic nicroalgae in use. Allhough very prornising It'sulis
alrd applications har,'e a]reacly bccu ucll estalrlisherl with such a small r]umber of
mlcroalgae, a much broacler variely irn(l l;rlger- nLrnbcLs of culture straits of
ruricroalg,ae are available from pLrblic culturc collections (...g. the SAG culture col-
l-"(:lion at (;ijttingcn Uni\,crsit)', Gcrnrany (lriedl ancl Lorcnz 20l2)) whir h. in addi
tion, 1nay be ever. expancled blr ncw isolales. Therefolt,, the vast rlivcrsit_v of
nricroalgac ancl nran,v aclclitionai rnii roalgal lincagcs still ren]ilin almost Lllttappcd
even though they providc vcr\r promising resources to be furthcr exploitecl. Many
obstacles still allparenl in microalgül lliotcchnoiog,y, e.g. those that rlelay thc clcvcl
opmL.nt of inexpensive lrcthods lor lliotnass produ(tion and thc optitlrization ol
compourcl acculnulation Ior using mir:roalgae irs solar drivc,n rellulirr fitctorics or
a source of bioluels may be solvccl lly thc cxtende(l and systernatic cxploitation ol
72 2 Phytogeny and systematics of mi'roalgae: An overview
-

used in or suitable for

Fig.2.1: Examptes of genera and species of microalgae currently
(a-d) cyanoba€teria. (a, b\ Atth tospito Fig. 2.2: Examples of genera ar
li-otecfrnologiial applkations. Fitamentous .platensis icat applications. (a) Vegetative
(u) coil"a fitaments and cells with gas vesictes (b) overview of
t"tr1ru11ro-i tt^r" iec el zq. (Chlorophyta, Trebouxiophycea
Filaments with rounded vegetative
colied filaments. (c) Anaboena catenula strain SAG 1403'l (f otmetly " Mymeci o" i ncisa ot
(arrow head) at the intercalary position (d) Oscillatorio soncto st(ain SAG
cells and heterocyte
Note the cross'walls within th€ fitaments (e) live cells of Eustignotos nagnt
74.79. Straight fiiaments without heterocytes
(Rhodophyta) from terrestrial ceae (Stramenopiles), with smi
norphyridium purpureum strain SAG 1380'1a, a unicellular red alga
appearance ln several cel[s' the centrally coccoid member of Eustigmato
trabitats, Note ttre reddish chloroplast with a star-like
green algat flagellate Hoemotococcus {een alga Chlorella vulgotis sl
located pyrenoid is visibte (f) Resting stages of the
that accumulated the red pigment the genus Crlorello. Note the s
pluvialis svain 192.80 (Chlorophyta, Chlorophyceae)
tetrathele sftain SAG761-2' empty sporangial celt walls (e)
astaxanthin. (g) Resting stages of the green algalflagelLate letroselmis
of lipid droplets scale bars: a' c' e' lariophy.eae, Stramenopiles).
(ciioropt \/tu,lltotoa"ndrophyceae). Note the accumulation
types (in the upper and lower F
f, 10 um; b, 200 Fm; d, 100 um.
SAG 2115 (Bacillariophyceae, S

view (centre), below the formal

SAG 926-1 (Haptophyta, Chron
 ,I rrrr.r LLr L tr ':1
-
- f
a, 1- t_! ',-n
' r]' 'l

t
,/
tI ,
I
| -*- ,

C 6

!ci
(:rt

',,D
S,S, '
..!,
q#,r/
-#, ,'f
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:r!1.2.t: lr,rrt)1r,..,. :r.,fi ,i,r rL.tre.lf! r rr r
i ( ri ) , . I :l , L r , . r i :r ll,i !!r( IIn e-rlIrll. i1' lr ir er lr rir Lr:

i- :r rph,rL,r. lflrrrrlrrlr(rl''l r,l l, .oIh . iLIIII I ,r!i s.rIirII!:r'( Lr) l e ( i'Ll "L,r l . i lr t Irr'(,"1
Llr) .rL,r',r1rlrr.,i. i'J isrr,, f.r,ir'lrrIlrrr. irrrrirr lr if,,rrrl] il!rlr lr'(l ir'rLtr' ll)lVu!.rr
r,,.' rr L ., r i iir. r r:,\li lr.3rr,,r rLr,'(,irl i.rh/-r.r'" rrerrbl .:I
irriririi.ri ,ii,,,t,ri|i ei
i!.:,: ,!. .'_rt.|. ) L€. . r,i1 !rrrll i, r )fi'rL! r.l \,r',i',,, i,1,,r.r!r, ii'ii'ri',1 'lrr r,r\(r I jr.:r irri| rrF
r!(oil |!|r,rL,;Lrt rt |,r rjiri !r :,rr 151r,r f;, !r1,.'rl : i r ( e ! r ' l r ,r I I r . . l ! l f L
, r L L I r , L 1 r , i r L lLil L

:',,r,riirriii,;r,lli,r'ri'r.,if',rlrr Srit. lll llhLl_i,r]]|l,r.rIrt)ii: )rr r!,!,'.r.'/l'r '.Irt,,')l

ll., trrrL,!l;,1(,,.lli []i]li'irIr il:iir':, rl i('lrL,i r,r 1, r.rr:'i, )rrrii I ir i:irrfr, l)!,i ,r ,L I
rftr'tr,rr,ir:,rt ,r r:L'.r,Lil. I ll.(rl l,r .llllrirr)ri,'rili,i'r,iri,.rrrilrirllri',r,r r!'1i;il'11, t a,,,',
.i,i. rhrrir,t.:,,r.:-.rij) .,!1.\(, r, I r,,l rr.nr_,r'rl!ritrlii,r(,reL !, Ie.'L: f! rr,r rtl ),.,ri i, ,ro
r,,!Llf lrL|)".r rr loi,r |,r i.r)l re r,rtr lllir,)rL',,i.:to_I/r'rr,i!!ii,ri,r'rL.r.'jli,ili] !Li,r I

(;\(, )il5i.l,iil ,1.,i ,r,.rf.rL.:irrIri,iLf',1i,."!',, lrr't'i:ir: r:l lLrilLirrrr' ;1lr lr", I'r
ri.r,i1rl r'1. r)e1,,,'; I iL'f,rl )r(,r fi-'...r'nii,r Lr)'Lr i('l.iitl\L'ilr,, i',r. !.r li!irr.' r,rilri"i
.,iLrr l,r'r'LL,L ,IIIL)!''I','Itr'!r,r'1:li: (llrr'
i, .l r I l: I |i,r. ( r i 'r r,,iL rlL" . i!.rl 'tr,.
r ) L I
1r '\ I ;r , )I r

74 2 Phylogeny and systematics of microalgae: An overview
-
Genus, species, taxonomic
position
Cyanobacteria
Aphanizonenon flos aquae
A rth rosp i ra (5 p i ru li n a)
Chlorophyta, Chlorophyceae
Ch Ia myd om on as re i n h a rdt i i
Dunollelld (two species)
H o e m ato coccu s pluvi o Lis
s ce n ed esm us/Desmo d esm u s
Chlorophyta, Trebouxiophyceae
Botryocaccus braunii
chlorella vulgaris
chlorophyta, chlorodendrophyceae
Tet rcse I m is (three spe.ies)
Rhodophyta
Porphytidium ctuentum
Bacitlariophyta (Stramenopiles)
choetoceros (four species)
Cy.lotella cryptico
Hasleo astreorio
Ph o eod a ctyl u m tri co rn
Tab. 2.1: Overview ol several species of rnicroalgae currently used in biotechnology: their
taxonomic position. growth characteristics, conrpound content and application.
-)-
Occurrence. growth Compounds and application Genus, species, taxonomic
position
Bacillariophyta (Strämenopiles
Freshwater Toxins SkclPtonena osrattln
High salinity and Pigments, high protein content
h igh pH
Freshwater Recombinant proteins
Carotenoids (ß-carotene), glycerol
Freshwater Fatty acids, carotenoids (Astaxanthin)
Freshwater Carotenoids (lutein), aquaculture L,laldssloslr, (four specles)
Freshwater oil, carbohydrates
Freshwater Fatty acids, carbohydrates
I\,\arine, brackish PUFA (EPA), d tocopherol, sterolsi Eustigmatophyceae (Stramenol
24 methylenecholesterol: cosmetics: Manodapsis subte onea
anti-wrinkles; aquacu{ture: rotifer feed, Nannochloropsis
shrimp hatchery. oyster and clam feed; (three species)
waste water treatment, cadmium
removal,
Rhaphidophyceae (Stramenopil
Marine, brac kish PU FA (EPA), pigments (phycoerythrin); Olisthadiscus sp.
and freshwater antiviral activitiy, sulfated polysaccha- Haptophyta (chromatveolates)
rides, antioxidants; poultry feed lsochrysis galbana
Marine, even at PUFA (EPA), lipids; p-E-gl!cans
h igh salinities, Povlova lutheri
thermophilic
Freshwater Lipids, B 1,3 glucan, bioacc!mulation
of metals; prophylaxis of myocardial
infarctl
Freshwater PUFA (EPA), linoleic acid; marennine
Dinophyta (Chromatveolates)
(colouring agent)i isoprenoids,
Crypthecodiniun rchnii
tetra-unsaturated Sestertetpenoids
(hastenes)
Freshwater. hetero- PUFA (EPA), lipid s
trophic growth
possible Eugtenoids (Excavata)
PUFA (DHA, EPA), lipids; p-carotene, Euglena grocilis
fucoxanthin;trace e[ements and
vitamins for human diet:
cosra etics
u tu n Freshwater, hetero- PUFA (DHA, EPA), lipids; aquaculture: Tab.2.1: (continued)
trophic growih rotifer feed
possible
the biodiversity of microal
study of growih condition
In parallel, a new appror
 2.1 Introduction _ 15

Genus, species, taxonomic

Occurrence, growth Compounds and apptication
position

Bacillariophyta (Stramenopiles)
Skeletonema castatLln
Marine PUFA tEpA)t antibacteflal r, ttvitV,
B-t,t g rrraf: ro<metic\. antj rg^rrg
af d anti-Lpllulite elfpLts, nqurr
ulrure
rnoltrscs and oyster fee;; ;;;,";";;
treatnrentr detoxification of caclmi!m
and copper, biodegradation of phenolic
cornpound5
Tholassiosirct(fourspecjes) Marjne
PUFA (DHA, EpA), lipiclsj carotenoids,
fucoxanthin; aquacLtlt!re: ieed
Ior
larvae. bivälves, oysters; waste
water
treatment, radmium rernovä1,
biodesra
datiof oI phcnoljc compoLnds
Iustigmatophyceae (Stramenopiles)
Ma n a d o ps i s s u b te rra ne a
Freshwater Most promising algal EpA producer
Nannachloropsis lMarine, brac ki5h PUFA (EpA, DHA), tipids; pismenls,
(lhree species)
.arotenordsr p t,l-qluLrnr pou
ny
fced: dquacrrlt!re: rotrfe ancj brvdlve
feed
Rhaphidophyceae (Stramenopiles)
Olisthadiscus sp. j\,4a
rin! tipjdsi B 1.3 glrr ca n
Haptophyta (Chromatveolates)
Isochtysis golbona Marine PUFA (DHA, EpA); sterols, alkenonei
aq!acult!re: clam ard oyster feed;
wasle waler lreatmenl
PUFA (DHA, EpA)j o tocopherot,
sterols
alkenone; cosmetics: arti-wrinklesj
aquaculturer rotifer, oyster and
clam
leed; shrjmp hatchery; cadmium
ren-iova I
Dinophyta (Chromalveolates)
Ltypthecodin i um cohn ii
Freshwater, hetero- High content of pUFAs (DHA); pigments
trophic s pec ies as colouring agents; extracis used
if
promoling lacti. acid and bilidus,
bacteria growlh: pharmaceuticals;
pet loods; aquaculture feed
Iuglenoids (Excavata)
Eugleno qtacilis
Freshwater, hetero PUFA, lipids; j ,l-g lu.
p- a r (paramylon),
trophic growth cI tocopherol
Poss ible
Tab.2.1: (continued)

the biodiver.sity of microaieae

\,!hich ntust also indude an expanded
study of srowth corditions lo obrair experimental
oprimat ),j"ld, ,rf ,ir;;;;i.;;i ärgat products.
In parallel, a new approach has 9q1u11r,d
wilh the succcsslul den.tonstralion of
 ---
76 2 Phylogeny and systematics of microalgae: An overview
-
genctically transformed microalgae, which may be optimized for the production of chemical path ways (e.g. aromr
Therefore, microalgae may be r
secondary metabolites by genetic engineering. This approach is currently furlher
explored in the dia tom Phaeodactylum (Fig. 2.2e), but also the green alga Chlamyd
omonas may constitute a promising alternativc systcm of gcneticall-Y transformed
microalgae. Genetic manipulation of microalgac in order to use them most effi- 2.2.2 Algal evotution
ciently and with new capabilities in biotechnological applications is sfill a vision
and associated with many problems (for a review, see Le6n (2007))' The mass An extensive reticulation occu
production of genetically modified microa)gae may carry many risks; for example' of endosymbiosis have shapec
the acccptability of protlucts from genctically modified organisms is often per- chine and Bowler 2011; fig..
ceived as problcmatic. ancicnt cyanobacleria lineage
rium by a hcterotrophic eukar'
ing and Palmer 2008; I(eelin8
2.2 Diversity and evolution of microalgae massive gene transfer from the
only a minor fraction of the p
2.2.1 Algal diversity whereas the maiority of plasti(
Palmer 2008; Fig. 2.1). Once
groups of
Thc biodiversity of microalgae is rcmarkable, The numbcr of eukaryotic with its host, thc three lineag,
microalgae is large, including at least fo taxonomic classes assigned to a variety
al. 2005: Tirichine and Borvler
of evolutionary lineages that are interspersed among thc protozoa, fungi, plants
(embryophytes), the ted algae
and anlnals. Eukaryotic life is seen to be distributed on at least five phylogeneti- plastida already represent a gl
(l(eeling
cally distinct "supergroups" as based on genome sequencing evidence cant, they represent only a fra
2004; Keeling et al. 2005); the algae are distributed on at least four of the five
acquired their plastids througl
recognized supergroups of eukaryotes (Keeling 2004; I(eeling and Palmer 2oo8i
rctention of a primary algal ce
Fig.2.1). It is estimated thatabout 3O,O0o 40,oOO algal species have been
2008). In most rases, the prir
described to date, but there is consensus that the number of still undiscovered was a red algal ccll (Fig. 2.1).
species may exceed known specics by a factor of tbur to eight
(Norton et a] 1996)'
for a third partner that has b€
Other authors estimate the number of actually existing algal species at 350'000' toms and presumably other l
with diatoms the most species rich group (Brodie and Zuccarello 2006)' For the grccn alga most likely precede
diatoms, the highest estimate is thai there may be thousands more species than firichine and Bowler 2011). {-
are currently described (11,000; John (1994)). Mlcroalgae are extremely abundant rachniophytes, are derived fic
and diverse in nature. For example, microscopic marinc phytoplankton are the The secondary spread of plas
primary photosynthetic organisms in the oceans, which cover over 70'/o of the
secondary end0symbiosis, th€
Earth's surface; here, their abundance almost always exceeds 10r' cells per liire progressively becn degenerat(
(Worden et al. 2004). Productivity of microalgae is very high' Whereas the biomass
until all that remained in mo
of vascular plants on land accumulates over time, singlc cell phytoplanl(ton in the
Because the endosymbiont \
ocean are quicldy eaten or killed by viruses, and so the slanding crop remains
host, secondary plastids have
more or less constant (Andersen 2O0B). Phytoplankton primary production the
so called complex plastids wi
amount of carbon fixed bV photosynthesis is estimated to be at lcast 40% of
mately connected to the nuc
global primary production (Bolin et al. 1977;MartinetaI 1987)'In other words' every
plants plastid endoplasmatic recticul
second breath we take concs fiom microalgae. Plastids are the olganelles of
process by biont is completely absent, t
and eukaryotic algae that harbour photosynthesis, which is the essential
representing the smallest kt
whichCOrisfixedtobuildupcarbohydratesandothercellularsubstancesusinglight
groups, thc Cryptomonads ($
encrgy. Plastids synthesizc many chemical compounds also important for other bio
 2.2 Diversity and cvolution of microalgae 17
-
chen]ical patlt ways (e.g. aromatic amilto acids. heme. isoprenoicls ancl fatty
acitls).
Thcreforc, microalgac nay bc regarcled as solar rlriven ccll factories.

2.2.2 Aleal evolution

An extensive reticulation occurred in algal evolution, i.e. rlair.rly two major c\,elts
of endosymbiosis have shapecl the algal trec of life (Bhattacharya et al. 2007;
Tiri
chine and Bowler 2011; Fig. 2.1). A so calieci ,,prirnary" endosymbiosis with an
ancielt (yanobacteria lineage, i.c. thc engulfn]ent and retention of a cyanobacte
riun by a lteterotrophic erlkaryotic ccll, nas lhe start of cul{aryolic algal lifc ((eel
ing ancl Palmer 2008; I(eeling tOlO; Iirichine ancl Bo.wler 2O1t). Subsequently, a
nassivc gene transfcr fr.m the endosymbiont to the host nLlcleus oc( urred. so rll.t
oniy a ntinor fraction of lhe plasijd,s proteins was still cncodcd by the organellc,
whereas the majority of plastid genes wcre transferrcd to thc nucleus (l(celine
ancl
i)almcr 2008; lig. 2.j). Once tlte endosymbiont ü,as established and integrateil
$'ith its host, the thrce lineages of Archacplastida (or plantae supcrgroup; (Acll et
a]. 2005; f irichjne ard Bou/ler 2011)) divcrged, i.e. thc grecn algae incl. lancl plants
(embryophytes), the red algae and the glaucophytes (Fi!.t. 2.1).
Although the Archac-
plastida alreadlr represcnt a great diversity anrl collectjvely arc erclogically
signifi-
cant, thcy represent only a fraction of eul<aryoli( phototrophs. Most algal
ljneages
acquired their plastids through secondary crrrlosymbiosis, lr,hich is the uptal<e
and
rclention of a primary algal cell b1r lne1h€,r-..Laryotic lincage (l(celing ancl palmer
20081. ln most cascs, thc primary algal cell involvecl in seconclary cnclosymbiosis
uas a red algal ccll (!'ig. 2.l). Howcver, rccently, phylogenomic evidence cnergcd
for a third partner thai has been i|volvcd iD the secondary enrlosynrbiosis
of dia
toms an{l ]lresumabl}, other hcterokont algae as \\,ell. An endosymbiosis with a
green alga ntosl lil<cly prececled the rcd algal cndosymbiont (Moustafa
ct al. 2009:
'Iirichine and Bolvler 2011). Otrly
two algal lincages, the luglenoicls ancl chlora
raahniophytes, arc derit'ed from a grecn algal seconclary enclosymbiosis (Fig.2.3).
'Ihc secondary spread of plastids
had a maior ilnpact on eukaryotic divcrsity. In
secondary endosymbiosis, tlle retained primary algal cell, lhe cnclosynlliont,
Ilas
progrcssivcly been rlegcneratcd including a massive endosynbiotic genc
transfcr
until all that renrained in n]ost cases was the plastid (l(celing and paim."r 2008).
llccause tlte eldosymbiont was insertcd into tlle clldomenlbral]e systcn of the
host, secondary plastids have one or t\,vo additional mel]lbrancs around
it fornting
so ca)led complex plastirls with three or lour'embranes and are somctimes inti
rnatelJr connected to thc nuclcus through a sharccl enclomembranc system,
thc
plastid endoplas1natic recticulum. I1.l most cases, the nucleus of thc algal
cndosym
biont is compietely abscnl, but a kind of relict nucleus (termecl nucleomorph),
representing tl'te smallest known eukaryotic genoules, persiste{:l in two algal
groups, thc Cryptomonads (wilh the plastid originatcd lrom a recl alnal
cell) ancl
78 2 Phvlogeny and systematics of microalgae: An overview
-
chlorophylls c such as th
e.g. the diatoms and bro\
phyta, as well as two al
qb
complex plastids and th(
Heterotrophic relict plastids (Fig. 2.3).
eukaryote 1
To make the reticulatr
phyte lineages even aban
algal cell and instead rec
)"1;- toms as their plastids (so

Glaucophyta
$o "io endosymbiosis may be fc
Secondary endosl.rnbiosir
at least once derived fron
Archaeplastida
or Plantae twice from unrelated gree
noids in the Excavata sul
ever, whether the Chrom€
group of eukaryotes is s
2010), while others consi
r'. creen alqa il alsa + Green is the Rhizaria may fall ,
g j,-:
Green
Heterotrophic ;; t'< q J'-i references therein). For a
eukaryote 2 "* t'""0",nu
++ i:-.
a robust and significant
already accumulated ovt
ii- - ,n'0
details still have remaine(
clearly demonstrates thal
I I I

Chlorarachniophyta two genomes, one from tl

Chromalveolates Euglenoids
Excavata Rhizaria intermixed through endor
have been so successful i
features, i.e. exhibit such
CrVptophyta Stramenopiles (heterokont algae): e g. be exploited by biotechn(
Haptophyta B a ci I t a ri o p hy ce a e ldialoms)
Xanthophyceae
Alveolates: Eustigmatophyceae
Dinophyta
Apicomplexa
Synurophyceae
Chrysophyceae
2.3 Cyanobacteria:'
Raphidophyceae
Phaeophyceae Cyanobacteria form a larl
Gram-negative bacteria. l
of eukaryotic algae, with which they s
Fig. 2.3: Schematic diagram summarizing endosymbiotic events and the evolution
atgat groups (adapted from Tirichine and Bowler 2011) Eukaryotic supergroups are in grey and broad variety of forms, ftr
bold tetters; algal ctasses and phyla discussed in the text are in bold letters; names of algal ated filamentous types to
are in
classes and phyla for which members are currently used in biotechnoLogical applications multicellularity (Kauff anr
italics.Thetwolevelsofendosymbioticevents,primaryandsecondaryendosymbiosis,arein are the oldest micro-orge
boxes.
to about 2.45 billion yearr
(Keeling and range, which makes ther
the chlorarachniophytes (with plastids arose from a green algal cell)
conditions. From an evolr
Palmer 2008; Keeling 2O1o). The maiority of secondary algal lineages belong to the
have been forced by the
Chromalveolates supergroup, which includes those algae with plastids that exhibit
---

2.3 cyanobacteriar The prokaryotic algae 19

-
chlorophylls c such as the photoautotrophic Stramenopiles (the heterokont algac,
e.g. the diatoms and brown algire (Yang et a1.2012)), the Cryp{ophyta, thc Hapto-
phyta, as well as two algal lineages of the Alveolates, i.e. the dinophytes with
complex lllaslids and the Apicomplexa, a paritsitic group with non'phototrophic
reljct plastids (Fig. 2.3).
To mal<e the reticulate evolution of microalgac cvcD morc complex, some clino
phyte lineages even abandoned their previous plastids that originated fiom a red
algal ce)l and instead recruited cells of eithcr cryptomonads, haptophytes or dia
toms irs their plastids (so called tertiary endosynbiosis ); even a third green algal
cndosymbiosis may be found in the Dinophyta (Gould et al. 2008; Kccling 2010).
Secondary cndosymbiosis is without doubt of several, i.e. at least three, origins:
at least once dcrivcd from a red algal endosymbiont, in the Chromalveolates, and
twice lrom unrclalcd grccn algae il.] chlorarachnio]rhytes in the Rhizaria and Eugle-
noids in thc Excavata supergrolrps (l(eeling 2004, 2010; I(eeling et al.2oo5). How-
cvcr, whether the Chromirlveolates actually represent a single monophyletic supcr
group of eul<aryotes is still being debaled by some auihors (c.g. Baurain et al.
2010), while others consider this supergroup of euharyotes to be even largcr, that
is the Rhizaria malr tall within Chromalveolaies (Tirichine and Borvler 2011, and
rcfcrcnccs thcrein). For all the endosvmbiotic events related lo plastid evolution,
a robust and significant wealth of genome or nultigene sequence cvidcncc has
alreacly accumulated over the pasi 10 years, even though scvcral aspccts and
details still have remained a natler of debatc; for a rcview, see l(eeling (2010). lhis
clearly demonslrates that the eukaryotic algae are of chimeric nature, i.e. at least
two genornes, one from the endosymbiont and thc other from the host, have been
intermixed through endosymbiosis. This may be the major reason why microalgae
have been so successful in evolution of life and are so diverse in their biochemical
features, i.e. exl.rlbit such an extremely broad variety of secondary metabolites to
be exploited by biotechnolo8y.

2.3 Cyanobacteria: The prokaryotic algae

Cyanobacteria form a largc and morphological diverse group of photoautotrophic
Gram-ncgalivc bacteria. Many cyanobacteria have large cells, like 1he eukaryotic
aigac, with which they share oxygenic photosynthesis. Cyanobactcria occLlr in a
broad variety of fbrms, tiom solitary unicells over color]y forming and undifferenti
aterl filamentous types to more complex forms that even display thc hallmarl<s of
multiccllularity (lGulf and llüdel 2011). There is strong evidence that cyanobaclcria
arc the oldest micro organisms to perfbrm oxygenic pholosynthesis, dating back
to about 2.45 billion years, Cyanobacteria exhibit a broad environmcntal tolcrance
range, which mal<es thenl well adapled to living undcr cxtreme and fluctuating
conditions. From an evolutlonary point of view, it appears that cyanobacteria may
have been forced by the "modcrn" cukaryotes to withdraw themselves into such
20 2 Phytogeny and systematics of microalgae: An overview
-
molecular nitrogen gas (N'z) into cyanobacteia can tolerat
habitats. Many cyanobacteria can fix atmospheric cent of an adaption to ea
amino acids and proteins' The
ammonia, which is then further assimilated into fer anoxygenic photosyn
process of nitrogen fixation is most often confined to
specialized cells' heterocytes'
key enzyme of nitro- bacteria are also very su(
which are capable to protect nitrogenase, the oxygen-sensitive their tolerance of desicc;
in the
g"n n"ation. However, nitrogen fixation can also occur in cyanobacteda Probably owing to th
protection are available'
ibsence of heterocytes if other mechanisms of nitrogenase the metabolic acitivities
I',litrogen-fixing cyanobactelia increase soil and
watel fertility and are palticulally
applications such as nut
fungi or being endosymbionts in
attraJtive as symbionts, e.g. to form lichens with ture as biofertilizer and ir
and diatoms (sharma et al' 2011) Cyanobac-
Oiu",t ,o*u.*uzoa, dinoflagellates and flocculants). ln addil
genera also chlorophyll b is present'
teria contain the chlorophylls a and d; in some olites with biological act
aswellascalotenoidsandphycobilinsasaccessolypigmentstocaptulelight tumoral and antiinflamr
inside the thyl-
.onu"y it to the photosynthetic reaction centres located years, cyanobacteda ha!
"n"rgy "nd pigments and are
akoiÄ membranes. phycobilins are blue or red
water-soluble their biomass, ethanol a:
present in large amounts. They are bound to proteins'
which form hemispherical ments, it is also potentiz
outer thylakoid surfaces' Several particles in tion of recombinant com
phycobilisomes, which occur on the
cyanobacteria may represent storage prodücts'.
ie' cyanophycin teria for various uses ha
ilr" .ytopturrn of
acids asparagine and arginine)' cyanophy- Classification of cyat
fu po'fvlnel of the nitrogen-rich amino
lipid droplets' carboxy- tionally been based on c
."ur].tur.t't 1u potyglucan) and polyphosphate' In addition'
somes (polygonal aggregations of nubisco, the
key enzyme in CO7 fixation) and and modes of propagati(
Gas vesicles (assemblies of hollow' bacteria are classilied in
,u. *ri.f", .un be present in the cytoplasm'
delimited by membranes) occur ftequentlv in aquatic
teriological code of nom
ioinr"a .nttnA"ir, but not Oscillatoriales, Nostocal
to confer vertical mobility
.yunob..t"riu and enable them to have buoyancy and botanical code of nomen
inthewatercolumn.ThecellwallofCyanobacteriaisbasicallythesameasthat cation schemes are knov
folGlam.negativebactelia,i.e.athinpeptidoglycanlayer(apolymelcomposed see, e.g. Sharma et al. (
cell membrane (cytoplasm
of sugar dedvatives and amino acids) is outside the not be sufficient to infe
layer is a space surrounded by another
-"rnirun"). Outside the peptidoglycan feature of cyanobacteria
relationships (l(auff and
membrane, termed the outer lnembrane' A charactedstic The cyanobacterial
sheath poly 1

is that they are usually surrounded by a mucilaginous -(extracellular cells lina Turpin ex Gomont;
which protects the
meric substances), mostly composed of polysaccharides' supplement; the Spiruii
from drying out and enables them to attach to substrates' filamentous colonies of
occupy diverse aquatic and
Cyanotlcteria are recognized for their ability to are used in China as a
variety of organic compounds
terresirial habitats, cyanobacteria produce a large cyanobacterium withoul
whichinnatuleareusedbyotherorganisms,andtheyStabilizesedimentsand the fact it has not beel
quantitatively important contri- Bornet & Flahault (as "l
soils. The cyanobactelia metabolic activities make
butionstothecarbon,nitrogen,sulphurandotherbiogeochemicalcycles'Bloom- amino acids, fatty acids
substances' termed cyanotox- to be viewed with caüti
forming cyanobacteria often release significant toxic
known from hot acidic springs that produce strong hel
ins (Sh"arma et al 2011). Cyanobacteria are also well
Many physiological proper- technological applicati(
ürougirort tfte -orld (e.g. Ward and Castenholz 20o2)'
to a large variety of extreme and those of the Andbaend l
ties tiat make the cyanobacteria able to adapt
long evolutionary history' Many Lyngbya C. Agardh ex (
environmental conditions may also indicate their (Fig. 2.1d). Surprisingly
cyanobacteria are able to tolerate low oxygen
conditions and even free sulphide at
most eukaryotic algae Many telrestlial biotechnological exploil
Ievels much higher than those tolerated by
 2.1 Cvärobacteria: The prokarvotic algae 21
-
- cyanobactcria can tolerate high UVB and C radiations, whicll may be also rcminis
cent of an adaption to early Earth's conditiolrs. Some cyal]obaclcria can even con
fcr anoxygenic photosynthcsis where IIrS is utilized as an elcctron donor. Cyano-
bacteria are also verl' successful primary colonizers in terrcstrial h.rbitats owing to
thcir tolerance of desiccation and to lvater stress.
Probably owing to ihcir high cap.rcity to adapt to almost all kjnds of habitats,
the metabolic acilivities of cyänobacteria also providc a broacl range of potenlial
appllcations such as nutrition (fbod supplcments and fine chernicals), in agricul-
turc as biofertilizer and in waste\{ater trcahnent (production of exopoiysaccharicles
and flocculants). In adclition, tl.rcy also produce a wide variety of sccondarl' netab
olitcs with biological activities, e.g. strorg antivjral, antibaclerial, antifungal, anti
tumoral and anti inflanllnatory activities, useful fbr therapeutic pttrposcs. ln recent
years, cyitnobacteria have gaincd interest for the production of biofuels, owing to
their biomass, ethanol and H: production. llecause of their simple growth require-
ments, it is also potentially cost eftedive to exploit cyanobacteria for tlte produc-
tion of recombinant compounds. An excellent rcvicw on the potential of cyanobac
leria for various uses bas been llresentcd by Sharna et al. (2011).
Classification of cyanobactetia above ihe lcvci of species and genera has trarli
tionally been bascd on cell organizalion, ability anil strategies of nitrogen fixation,
and modcs of propagiition (l(auff and Büclcl 2o1i). ln current sr:hemes, the cyano
bactcria are classitied into five subsections (Subsections I V) according to the birc
tcriological code of nomenclaturc, and five orders, Chroococcalcs, Pleurocirpsales,
Oscillatoriales, Nostocalcs and Stigonematales, i{hich havc bccn erected under the
botanical code of nomenclature. Several of the groups dcscdbed by current ciassifi-
cation schemes are knorvn to be polyphyletic in molecular ph1'logenies (for reviervs,
see, e.g. Sharma et al. (2011)), and phylogenies bascd on only a sirgle gene may
not be sufflcient to infcr a clranobacteria classification that reflects phylogenetic
relatlonships (l(auff and 1lüdel 2011).
Thc cyanobacterial genus ,4rrhrospira Sitzenberger ex Comont (formcrllr s'plru
lina Turpin ex Gomont; without hctcrocytes; Fig. 2,1a,b) ls ttscd as hurnan food
supplement; the Spirulind industry is of increasing econonic inportance. Large
filamentous colonies of Nostoc Vaucher ex llornet & Flahault (with heterocytes)
arc used in China as a "hair vcgetable" (Facai). filamcntous freshwater
^nolher
cyanobacterium \a,ithout hetcroclrtes even being used as a food sLLpplement cl-"spite
the fact it has not bccn licencecl as such is Aphanizonenon flos c4uat: Ralfs ex
llornet & Flahault (as "AFA' alga). Although it provides a high content of essential
amino acids, fattl' acids, minerals and vilamins, its use as food supplelt.lcltt needs
to be viewed with caution because the spccies is knolvn fot fotming algal blooms
that producc sirong hepatotoxins and inhibitors of nerve functiolls. In othcr bio
technological applications, additional filamentous ryanobacteria are used, c.g.
those of the Anabaena Bory ex Bornct & Flahault (with heterocylcs; Fig. 2.1c), the
l,yngbyu C., Agardh cx Gomont an.l OsciLlaloria morphotypes wjthout heterocytes
(Fig.2.1d). Surprisingly almost no uniccllulat cy.rnobacteria have been uscd for
biotcchnological exploitation, even though they also can be easily grou,n.
 ---
22 2 Phylogeny and systematics of microalgaer An overview
-
2.4 Plantae or Archaeplastida supergroup: Green algae, The green algae have
red algae and glaucophytes Wodniok et al. 2011; Fde(
phylum Chlorophyta, cor
The green algae and the embryophytes (multicellular land plants) constitute the algae in a more narrorv sr
Viridiplantae, which, together with the red algae (Rhodophyta) and the glauco green algae, with a bewi
phytes (Glaucophyta), form the "Plantae" or' "Archaeplastida" supergroup of eul<ar- different morphological c
yotes (Adl et al. 2OO5; Keeling 2010). They are the only photosynthetic eukaryotes eage, the phylum Strepti
with double membrane bound plastids; they are derived from primary endosymbi cally rather simple line:
osis. The phylum Rhodophyta is often regarded as a sister group to Viridiplantae, organization (Becker and
and the Glaucophyta is considered as the most basal lineage. 1he monophyletic Lauterborn and the sarcir
origin of has been supported by nuclear and chloroplast multigene ing lineage of the Strept(
analyses ^rchaeplastida
(Rodriguez-Ezpeleta et al. 2005; Archibald 2OO9). est living relatives of emt
recent analyses of nucle
however. revealed that ei
2.4.1 Viridiptantae: The green atgae distributed over two phyta or a group unitins the Z
group to embryophytes (\
Features that set apart the green algae (as well as embryophytes) from other earlier divergence and no
Archaeplastida are that their double membrane bound chloroplasts exhibit stacked of streptophyte green algr
thylakoids, and that they contain the accessory pigment chlorophyll D. The other likely owing to ihe fact th
accessory pigments are the caroienoids beta carotene and xanthophylls, The main compounds in these alga
reserve polysaccharide is starch, which is deposited inside the plastid (Lewis and closely related to land pli
Mccourt 2OO4; Friedl and Rybalka 2012). A few genera of green algae contain even compounds otherwise sir
non photosynthetic plastids without chlorophylls, e.g. Prototheca W lCüger and plants. The polyunsaturi
the parasitic Helicosporidium I(eilin and, therefore, grow heteroirophically in the found to be more frequi
dark. Most of these characters are shared with embryophytes (land plants), and group, with relatively hi!
therefore, apart from being mostly not of true multicellular organization (l-eliaert genus Closfe,"ium (Lang e
et aI.2012) and often confined to aqueous habitats, it is difficult to delimit the ceans, members ol the ot
green algae from embryophytes without considering their very diverse gross mor extracellular polysacchar
phologies (Friedl and Rybalka 2012). Green algae vary morphologically from the The Chlorophyta con
smallest known eukaryote (the prasinophyte Ostteococcus Courties & Chret. Dinet) lineages, i.e. the classer
and tiny flagellates to giant unicells with multiple nuclei or multicellular forms Chlorodendrophyceae, fo
reminiscent of bryophyte gametophytes (Coleochaete Brebisson). The majority of early offspring, an asser
green algae thrive in freshwater or terrestrial habitats, but some microscopic forms prasinophyte algae (Fried
(prasinophytes) are abundant in marine phytoplankton. It is typical for green algae Members of Chloropl
to live in various terrestrial habitats (Holzinger 2009). Among green algae, there and there are only very fe.
are many "land plants," i.e. transitions to the land happened many times in the of unicellular or colonial
evolution of the green algae (Lewis and Lewis 2005; Lewis 2007). Examples of naeus) and many membe
terrestrial green algal life include the biofilms of building facades, biological soil or branched (e.g. Str.qeor
crusts (Büdei et al. 2009) or algal crusts on trees. Excellent reviews on the peculiar dividcd into five clades t.
ities of terrestrial green algae have been presented by L6pez Ilautista et al. (2007) Two of these clades, Spha
and Rindi (2011). Green algae, especially a variety of members of Trebouxiophy ydomonadales), are closr
ceae, are frequently involved in symbioses with ciliates and metazoans. widely used for biotechnr
 2.4 Plantae or Archaeplastida supergroup 23
-
-_ The green a)gae have evolved in two major lineages (Lewis and Mccourt 2004;
Wodniok et al. 2011i Friedl and Rybalka 2012; Leliaert et al. 2012). One lineage, the
phylurn Chlorophyta, comprises the majority of green algal diversity, i.e. the green
algae in a more narrow sense or the majority of what have been traditionally called
green algae, with a bewildering array of morphologies attributablc to at least five
different morphological organizatjons (t'röschold and Leliaert 2007). The other lin-
eage, the phylum Streptophyta, comprises no more than four or fivc morpholttgi
cally rather simple lineages and two advanced lineages with true multicellular
organization (Becker and Marin 2009). A lineage uniting thc flagellate Mesostigma
Lauterborn and thc sarcinoid Chlorol<ybus Geitler may represent the earliest divcrg
ing lineage of the Streptophyta. Which streptophyte algal Sroup may bc the clos
est living relatives of embryophytes has been heavily debated in recent years. Most
recent analyses of nuclear genes as well as chloroplast phylogenomic ana)yses,
however, revealed that either the Zygnematophyceae (synonym Conjugatophyceae)
or a group uniting the Zygnematophyceae lvith Coleochaetophyceae is the sister
group to embryophytes (Wodniok et aI.2011). The Charophyceae is now seen as an
earlier divcrgence and not as the closest relatives of land plants. No single member
of streptophyte green algae is currently used in biotechnological applications, most
likelv owing to the faci that no study has been carried out yet to screen for valuablc
compounds in these algae. However, because the streptophyte green algae are so
closelv related lo land plants, they may be particularly interesting for prcctlrsor or
compounds otherwise similar to those so far only known from multicellular land
plants. The polyunsaturated fatty acid (PUFA) y linolenic acid (GLA) has been
found to be more frequent in streptophyte green algae than in any other algal
group, with relatively high percentages in some strains of the zygnematophycean
genus Clost€riurn (Lang et ai.2011). other rather small unicellular zygnematophy
ceans, members of the order Desmidiales, are known to grow fast and to produce
extracellular polysaccharides that may be interesting to explore further.
The Chlorophyta comprises the vast majority of green algae, with four major
lineages, i.e, the classes Chlorophyceae, Trebouxiophyceae, Ulvophyceae, and
Cblorodendrophyceae, forming the "crown group" of Chlorophyta and, as their
early offspring, an assemblage of various monophyletic lineages of unicellular
prasinophyte algae (Fdedl and Rybalka 2012).
Members of Chlorophyceae thrive mostly il.l freshwater or terrestrial habitats,
and there are only very few symbiotic genera. The Chlorophyccac includes a variety
of unicellular or colonial flagellates (e.g. Chlamydomonas Ehrenberg, voLvox Lin'
naeus) and many membcrs with firm unbranchcd (e.g. Oedogonium Link ex Hirn)
or branched (c.g. Sligeo.lonium l(ützing) filaments. lhe Chlorophyceac may be
divided into five clades that arc robustiy resolved in most phylogenetic analyses.
Two of these clades, Sphaeropleales and Volvocales (sometimes also called Chlam'
ydomonadales), are close relatives to each other; they contain green algae most
widcly used lbr biotechnological exploitation: the coccoid colony foLming genera
 t,-
24 2 Phylogeny and systematics of microalgae: An overview
-
Chlorella sPecies has been
Scenedesmus Meyen and Desmodesmus (Chodat) (R Choda0 S S An' T Friedl &
Hae Several former " Chlorella"
E. Hegewald. (Sphaeropleales), the green flageltated geneta Chlamydomonas'
(Volvocales) IJoth genera' roidium Nadson, Parachlc
matococcus l"lotow (Fig. 2.1f) and' Dunaliella Teodoresco'
to the Sphaeropleales and are clearly Huss, T. Rohr & M. Wolf'
Scenedesmus and Desmodesmus, belong
Silva. C. pyrenoidosd H. c
separated from each other by their cell wall features ln Desmodesmus'
certain
cell wall vulgaris. C. zofrngiensis D
submicroscopic structures (ornamentations) are present on the outermost
are in heen found to be member
layer and one or several spines are on each cell Those structures absent
of Dunaliella chloris and Graesiella (Gt
s.encdesmu.s. and onlv rib-like structures may be formed' Species
W. Itüger, whlch exhibits
a llneage of their own within the Volvocales (Nakada et al 2008)'
The astax-
form
relative of the "true" Chlo
anthin-producing Haematococcus (Fig. 2.1f) is within a distinct tineage of
Volvo'
dilferent hom Hae minute and fast growing
cales. which also includes a variety of algac morphologically
(with a high relative freshwatel PhYtoPlanktor
matococcus, e.g. the coccoid green Chlorococcum Meneghini
logical Potential. Two mc
DHA content (Lang et al. 2011)) and Efrli( Komärek (Nakada et al 2o0B)
The latter
which was found to exhibit an nological Potential are m
genus also comprises "Neochioris" oleoaburuLans,
are species of ryococcus br.lunii Kitzin
extraordinarily high oil content. Close relatives of Haematococcus
a red pig hydrocarbon fuels (Weis
the flagcllate Chlorogonium Ehrenbcrg, which were found to accumulate (Reisigl) IGrsten, Friedl
well, probably astaxanthin (J' Ftedersdorf and M
ment in lag phase cultures as
are distributed on sevetal iinea- incisa Reisigl ar]d Pariett
Lorenz, pers. comm.). Species of Chlamydomonas
genera (Nakada et as the richest "Plant" so
ges of the Volvocales, which have to be attributed to different
2002b).
ut. ZOO8). The type species C. reinhardtii P A. Dangeard is, together with
morpho'
its own (Nakada et al' The UlvoPhYceae is
logically different flagellated green algae, in a lineage of
(formerly Chlorophyta (Leliaert et
200B). Other green flagellates, species of Tetraselmis F Stein Pla4lmonas
and marine: it includes all m
G. S. West; Fig.2.1g), which are used in waste water treatment' aquaculture
to the those with multicellular
cosmetics production, are not members of the Chlorophyceäe but belong
of the four maior Chloro- tubular forms as UiYa L.
class Chlorodendrophyceae, which is the most basal
les is another unique lil
phytan lineages (Friedl and Rybalka 2012)'
forming filaments of un
For the class Trebouxiophyceae, tbe majority of presently known
members are
occur' and a few lineages mulate P-carotene when
coccoid unicells; in some lineages also colonial coccoids
may also form filaments. Flageilated vegetative forms are not known for the class'
Members of Trebouxiophyceae are mostLy found in drier habitats' e
g in soil' or
phytoplankton (e g 2.4.2 Rhodophyta: Red
are aerophytic atgae. Many lineages include minute freshwater
Chlorellales). There are numerous examples for symbioses in ciliates' metazoan
Red algae are a very iar
(Pröschold ei al. 2011) and lichens (Friedl and Büdel 20Os)' Very few members of
the most (Brodie & Zuccarello 20
Trebouxiophyceae are currently used for biotechnological exploitation;
(Fig 2'2d) and a few more Chlorella' importance. Though tht
prominent may be Chlorella vulgaris Beiierinck
ity of red algal genera t
Iik" gru"n algue. The genus Chio,"elia in its traditional circumscription is of multiple
the play important ecologi
origins and distributed over at least two classes of green algae' An overview of
of the type species' C vuigaris' (e'g C' importance are species
"ftle" Chlorella species, i.e. close relatives are grown in maricuitu
of
sorokiniana Shihira & R. W Iftauss) and other members of the Chlorella-clade
genera are cultivated I

theorderChlorellales,hasbeenplesentedbyLuoetal.(2010).WithinC.vulgaris'
such as agarose and ca
a remarkable genetic diversjty has been found among va ous isolafes
of the same
history of chlorophYll b and c' br
species (Müller et al.2OO5). An ovetview of the confusing taxonomic
 2.4 Plantae or Archaeplastida supergroup 25
- -
Ch lorella species has been presented by Huss et al. (1999) and l(rienitz et al. (2004).
Several former "Chlorella" species have been transferred to other gencra, e.g. Chlo,
rotdium Nadson, Parachlorellct L. I(rieniz, E. H. Hegewald, D. Hepperle, V A. R.
Huss, T. Rohr & l\4. \ /olf, a\d Watanabea N. Hanagata, I. I(arube, Chihara & p C.
Sjlva. C. pyr"cnoidosd ll. Chick is regarded as a synonym with the type species, C.
vulgaris. C. zofingiensis Dönz and C. fitsca var. vacuolata Shihira & Krauss have
been found to be members of Chlorophyceae and transferred to the g enera Chromo-
chloris and Graesiella (cuiry and cuiry 2012). The heterotrophic protothccq zopfii
\{,/. Krüger, which exhibits a high relative DHA content (Lang et al.2O1t), is
a closc
relative of the "true" Chlorella species. The order Chlorcllales comprises many more
minutc and fäst growing coccoid green algae (e.g. Nannochloris Naumann) from
frcshwater phytoplankton, for rvhich it would be interesting to test their bjotechno-
logical potential. Two morc coccoid green algae well kno.wn for their high biotech
nological potential aIe members of two distinct lineages of Trebouxiophyceae: ttot,
ryococctLs braunü Kützing which is regarcled as a promising source of renervable
hydrocarbon fuels (Weiss et al. 2{J10; Tanoi et al. 2011) ancl Lobosphaera incisa
(Reisigl) I(arsten, Friedl, Schumannn, Hoyer & Lembcke (synonyms Myrmecia
ii?cis.r Reisigl and Parietoc,iiloris incisa (lt. Reisigl) S. Watanabe; Fig.2.2a) known
as the richest "plant" source of the PUFA arachidonic acid (Bigogno et al. 2002a,
2002b).
The Ulvophyceae is probably morphologically the most diverse group of the
Chlorophyta (l.eliaert et al.2012). The class may be regarded as predominantly
marine; it includes all macroscopic marine representatives of the Chlorophyta, e.g.
those with multicellular bodies composed of uninucleate ceils, forming blades or
tubular forms as Lllva L,, which is known to form ,,!ireen tidcs,'. l.he Trentepohlia
Les is another unique lineage of Ulvophyceae with exclusively terrcstrial
members
forming filanents of uninucleate cells. They are known for their capacity to accu_
mu'arr p-r aro,en(. irhcn cxpnsed ro air.

2.4.2 Rhodophyta: Red algae

Red algae are a very large and diverse group of nicroscopic algac and macroalgac
(Brodie & Zuccarello 2OO6). They are best kno$/n for their economic and ecological
inportance. Though ihey are present in freshwater with scveral genera, thc major,
ity of red algal genera occur on tropical and temperate marine shorcs, where they
play important ecological roles (Graharr et al. 2009). Of rvell-known economrc
importance arc species of Porphyra C.Agardh and other rccl algal species, rvhich
are grorrvn in mariculture operations for use as human food. Several other marine
genera are cultivated or harvested for the extraction of gelling polysaccharides
such as agarose and carrageen (Grahan et al. 2OO9). The plastids of red algae lack
chlorophyil b ancl c, but contain phycobilins (allophycocyanin, phycocyanin, and
 26 2 Phylogeny and systematics o[ microalgae: An overview 2.5 Chromalveol
-
the red pigment phycoerythrin) as accessory pjgments, which are located in phy pigments, fucoxant
cobilisones on the outer surface of the thylakoids. Red algae produce floridean (except in eustigma
starch, which is depositcd in the cytoplasm. Ihe rhodophytes lack flagella and endoplasmatic reti(
centrioles in all stages of the lifc history (craham et al. 2oO9). The Rhodophyta connected. The pho
conprises only few microscopic forms. The unicellular species of porphyridium cal diversity: a largr
Nägeli (Fig. 2.1e) are well known for their high content of polysaccharides as well with silica fiustule:
as rich sources of pigments and some pUFAs, e.g. EpA. High relative LpA contents phytes) as wcli as t
have also been detected in the multicellular, but more simpJy organized Conpso- at sea shores. Base
pcrgonopsis leptctcladus (Montagne) V l(rishnam. arLd Actochaetium virgatulum and molecular phyl
(Flarvey) Batters. Stramenopiles have
recent rcview of ph,
opiles (heterokont a
2.4.3 Glaucophytes opiles that are currr
rescnt the diatoms,
Glaucophytes (or glaucocystophytes) form a small group of microscopic algae
exclusively found in freshwater environments; only about ll species of glauco_
phytes have been described so far. Glaucophyte plastitls are unique among plastids
2.5.1 Diatoms (Baci
in that they are retaining the prol(aryotic peptidoglycan layer between ther'r two
membranes. Molecular phylogenetic analyses shorv thc glaucophytes to be the ear_
Diatoms are unicelli
liest divergence within the plant supergroup (KcelinS 2010). Glaucophyte plastids
recognizable algal g
share the accessory photosynthctic pigments, phycobilins, which are otganized in
cell walls, termed ft
phycobilisomes (small particles on the outer surfäce of thylakoid membranes) with
pattcrns of ornamer
cyanobacteda and rhodophytes. So far, glaucophytes have not been used for bio
sist of two parts, D
technological exploitation. However, a high frequency of Ep containing strains
brown carotenoid fr
has been found among claucophytes (Lang et al.2011), and some species, e.g.
biosis. Othcr chara
Cyanophora paradoxa I(orshikov, may also be easily grown in mass culture.
There are two mäjo
radial symmetry in
2.5 Chromalveolate algae: The photosynthetic Stramenopiles i.e. their gametes all
(heterokont algae) rliaioms have elongr
with a raphe exhib
Stramenopiles are one major group of the Chromalveolates supergroup of eukary_ isogamous, i.e. thei
otes (l(eeling et al. 2005; Tirichine & Bowler 2011). l hey form a monophyletic group diatoms are often r€
of photoautotrophic as \,vell as heterotrophic organisms, which are characterized phyceae; e.g. Yang e

by swinming cells posscssing at least onc flagellum with distinct tripartite tubular the diatoms the ranl
hairs (Andersen 2004; Grahan et al.2009). Most Stramenopiles have two distinctly three classes: the tr
different flagella and, therefore, are also known as heterokonts (heierokont algae; diatoms, whereas th
Heterokontophyta). I'hey have a long f'lagcllum with tripariite hairs (tinsel or imma_ I{aczmarska 2004). '
ture flagellum) to pull the cells through the water and a shorter smooth onc (whip- bers in the tieshwa
lash or mature flagellum) that lacks tripartite hairs, often with a light sensing fla dominantly in the n
gellar swelling at its base (Andersen 2004j craham et al.2009). photosynthetic Diatoms are pro
Strameüopiles usually appcar brown or golden brown (and are thus sometimes mately /+0 % of the
referred to as chromophytic algae owing to thc presence of charactedstic accessory and these are respc
 2.5 ChromaLveolate algae: The photosynthetic Stramenopiles (heterokont
algae) _ 27

pigments, fucoxanthin or vaucheriaxanthin,

and at ieast one form of chlorophyll c
(except in eustigmatophytes). In most photosynthetic
stramenopiles, a chioroplast
endoplasmatic reticuium is present, i.e. the plastid
and nuclcus are strllcturally
co[nected. The photosynthetic Stramenopiles comprise
a tremendous morpho]ogi
cal diversity: a large variety of unicellular or
colonial algae, including the diatoms
with silica frustules as walls, multicellular simple filaments (e.g.
in the xantho_
phytes) as weli as the macroscopic
complex brown atgae iorm;ng a brown canopy
at sea shores. Based on detailed analyses of pigment
compositi"on, ultrastructure
and molecurar phyrogenies, more than a dozen
differeDt classes of photosynthetic
Stramenopiles have been described (Andersen
2004; Graham et al.2009). For a
recent review of phylogenetic relationships of
lineages of photosynthetic stramen_
opiles (heterokont algae) see Yang et at. (2o12).
elasses of phorosynthetic Stramen
opiles that arc currently Llsed in or explored for
biotechnological applications rep
resent the diatoms, the Eustigmatophyceae and
Raphidophyceae.

2.5.1 Diatoms (Baci[ariophyta; photosynthetic

Stramenopiles)

Diatoms are unicellular or colonial eukaryotic algae

and are one of the most casily
recognizable algal groups because of their unique
architecturaily comilrex siliceous
cell walls, termed frustules. They are composed of
amorphous silica with regular
pattems of ornamentation. extensivcly used
as taxonomic ,.fingerprints,,, and con_
sjst of two parts, Diatoms contäin brown plastids
owing to t"he presence of the
brown carotenoid fucoxanthin and are derived from
red Jgal ,".oi,luru endosym-
biosis. Other characteristic accessory pigments are
'l'hele ale two major
the ihtorophylls c1 and c2.
types, centric and pennate diatoms. centric äiatom,
hau" u
radial symmetry in valve view, contain severai small plastids
and are ooeamous.
i.e. their gametes are an egg and motile spermatozoids
with one flagerum. pennate
diatoms have elongated cell walls with bilateral
symmetry. those ierrnatu ,tioto_.
witlr a räphe exhibit gliding motility. They contain one
or few plastids and are
isogamous, i.e. their gametes are of equal shape
and without flqqella. While the
diatoms are often regarded as a class of photosynthetic
.t.",nonoiil., (Bacillario,
phyceae; e.g. Yang et al.20r2), a more
recent taxonomic afiangen]ent even assigns
the diatoms the rank of a phylum, Bacirlariophyta.
The latter is further divided into
threc classes: the Mediophyceae anrl Coscinodiscophyceae
comprise the ccntric
diatoms, whereas the Bacillariophyceae encompass
all pennate forms (Medlin anrl
IGczmarska 2004). While peonate diatoms ur" r"pr"r"ntod
in roughly eqLral num
bers in the freshwater and marine habitats, the
centric diatoms pr"r"r,, pro_
dominantly in the marine environnlent (Lee 2O0S). "r"
Diatoms are probably thc most species rich group
of eukaryotic algae. Approxi_
matcly 40 % of the global annual marine bionass production
is due to diatoms,
and these arc responsible for about 25% of total terrestrial
primary productjon,
28 2 Phytogeny and systematics of microalgae: An overview 2.5 Chromalveolate a
-
making them the most dominant group of organisms sequesteting carbon from the eign DNA into diatom (
atmosphere (Bozarth et al. 2009). They may be the most important group of eukary- ments of the diatom cel
otic phytoplankton in marine and freshwater environments but are also abundant genome which would a
as benthic forms, growing on sediments or attached to submersed substrates, and (Bozarth et al. 2009).
can also be found in soils. Diatoms form important symbioses with nitrogen-fixing expression factors has
cyanobacteda in tropical seas. Diatom frustules predominate in the sediments of naturally produced by
the ocean floor, and the analysis of diatom frustule containing sediments can pro, successfully introduced
vide information on past environmental conditions. Diatoms produced significant granules in the cytosol
fossil records, and their fossil deposits are even mined and used commercially. used successfully to pro
Diatoms are considered to provide enormous economic potential (Kroth 2OOZ atitis B surface proteil
Bozadh et al. 2009). They are a valuable source for the acquisition of food supple- C02-neutral compared\a
ments (e.g. because of their high content of vitamins and amino acids) as well as or mammalian cells (He
substitutes for synthetic substances (e.g. cosmetic chemicals) and produce antibi-
otics and pharmaceutically active substances. Diatoms are a rich source of pollun-
saturated fatty acids (PUFA), especially eicosapentaenoic acid (EpA), arachidonic 2.5.2 Eustigmatophy(ei
acid and docosahexaenoic acid. Other industrial applications considered and initi-
ated for commercial use of diatoms include renewable energy, e.g. ethanol produc- Despite being only disti
tion via fermentation, and methane production as well as fluid fuel production. heterokont algae, Xanth
Their unusual cell wall offers prospects for applications in nanotechnology and similar, and they are al
computer chips. Therefore, a number of diatom species have already been exploited classes lack fucoxanthil
for biotechnological applications, also due to their easy growth. There are even yellow- greenish coloratir
species that can grow heterotrophically (Perez-Garcia et al. 2011) and even without to certain green algae,
the addition of silica to the growth medium. A centric diatom (Coscinodiscophy- levels occur within the r

ceae) used for the production öf PUFAS is the fteshwater species Odontella aurita previously been descrit
(Lyngbye) C. Agardh (Pulz and cross 2OO4). Other Coscinodiscophyceae are used Eustigmatophyceae, onl.
in aquaculture, e.g. as a food source for shrimp lawae and bivalve mollusk postlar- Eustigmatophyceae even
vae, i.e. the marine Skeletonema cosfatum (Greville) Cleve, marine species of Chae- heterokont algae), and tl
toceros and, Thalassiosira (Fig. ).2f). Cyclotella ctyptica Reimann, Lewin & Guillard ence of which is a chari
(Coscinodiscophyceae) has been used for silica production (Csögör et al. 1999) and of unicellular coccoid E
as a source of PUFA for aquaculture; the species can also be grown heterötrophi- applications. Species of
cally (Pahl et al. 2010). Further examples of diatoms capable of heterotrophic sqltna D. I. Hibberd; Fig
growth are the pennate Phaeodactylum tficomutum Bohlin (Bacillariophyceae; Fig. (N. Iimnetica lftienitz, t
2.2e), Nitzschia aiba J. C. Lewin & R. A. Lewin and Nitzschia laevis Hustedt (Wen subterrqneus J. B. Petersr
and Chen 2003; Pahl et al. 2010; Perez,Garcia et al. 2011) When grown autotrophi- PUFA EPA (Cohen 1999),
cally, P. tricornutum can grow even independent of silica. The pennate diatom later was found to be a
Haslea osfteatia (Gaillon) Simonsen contains the hydrosoluble pigment marininne transfeffed to a new gel
and, therefore, is used as a food source in aquaculture for oyster greening (Lebeau The position of M. surfel"
and Robert 2003a, 20O3b). matophyceae has been p
Genome projects are regarded as promising milestones for the effective genetic terranea ls closely relatr
engineering of diatoms. Complete genomes are already available for the pennate oculata, N. limnetica) whj
species P. tricornutum (Fig.2.2e) and lhalassiosira pseudonana. Transformation logical exploitation. Oth
protocols have already been developed which lead to the stable integration of for- described as members o
 2.5 Chromalveolate algae: The photosynthetic Stramenopjles (heterokont algae) 29
-
eign DNA into diatom cluomosomes. Future goals are to target differe[t compart
nlents of the cliatom cell to jntroduce new capacities and also transform lhe piastid
genone which would allow tlte expressiol.l of foreign genes of prokaryotic origin
(Bozarth et al. 2009). Thc great potenlial of diatoms to serve as solar_powcrerl
expressiot] factors has recently becn demonstrated. The patltway of a polyestcr
naturally produced by certain bacteria, poly lhydroxybutyrate (pHt3), lias bcen
succcssfully introducecl jn p, cornutum, which has produced bioplastic as small
granules in the cytosol (Hempel et al. 2011a). l.urtltelmore, p cornutum
has been
used successfully to producc a recombinant protein (human antibody against Ilep-
atitis B surface protein) through photosynthesjs with the ailvantage of being
COr-neutral compared with cxlllession systems used so far, such as bacteria, yeasts
or mammalian cells (Flempel et al.20tib).

2.5.2 Eustigmatophyceae and Xanthophyceae (photosynthetic Stramenopites)

Despitc being only distantly related within the Stramenopiles, the two classcs of
hetcrokont algac, Xanthophlrceae ar.rd Lustigmatophyceae, may be phenotypically
similar, and tltey are also linked by thcir taxonomic llistories. Members of bolh
classcs lack futlxanthin, and because chlorophyll b js also absent, they have
a
yello',v grcenish cololation. XaDthopl.iyceae are r.ather sinilar ill their gror{th
lorms
to certain grecn algac, i.e. coccoid, filamentous ancl siphonous organizational
levels occur within the class as in Chlorophyceae. ln fact, sonc green algac have
prcviously been described as mcmbers of XanLhophyceae and vice vcrsa.
For
Eustigmatophlrcqne, only coccoid members are l<nown so far (piibyl ct al. 2012).
llustigmatophyccae even lack chlorophyll c (the only known exception within
the
hcterokont algae), and their chloroplasts do not contain a girdle lamelJa, the prcs-
ence of which is a characteristic feijtLlre of most stramenopile algae. Trvo gcnera
of unicellular coccoid Eustigmatophyceae are curently useci in bioiechnological
applications. Spccies of Nnlnochloropsis D.J. Ilibberd occur in both marine (N.
salina D. I. Hibberd; Fig. 2)c: N. o.ulatd (Droop) D. J. Hibberd) and freshwater
(N. Lintnetica lftienitz, D. llcpperle, H. B. Stich & W Weiler) habitats.
suhterrant:us J. lJ, Pctersen, regardecl as the most promjsing algal producer ^4oi?odus
of the
PUFA FIPA (Cohen 1999), has been dcscribecl as a membcr of Xanthophyceae,
but
later was found to be a mcmber of Lustignratopltyceae and, thercfore, has becn
transfeued to a new gcnus as Monodr;p sis subterrunea (J. B. petersen) Hibberd.
The position of M. suhterranea as lvell as another species of,l4onodus in the
lLLStig
matophyceae has becn provcn b)'nolecular phylogeny (I,iibyl et al. )O12). M. sub-
tetranea ls closely related to species of Nannoc:hlorctpsis (N. salina; fig. 2.2c; N.
oculata, N. limnetica) \vhi(:h are among the most pollular microalgae for biotcchno_
logical exploitation. Othcr uniccllular coccoid yellow grcerrish algae, currcntly
described as mcntbers of XaDthophyceae, ntay actually be founcl as rrembers of
 t,-
30 2 Phylogeny and systematics of microalgae: Ar overview 2.6 Chromalve

-
Eustigmatophyceae after a closer pigment analysis and molecular phylogenetic ment. In contrast to most di
investigaiion (Piibyl et ai. 2012). Their biochemical properlies may even be similar tion also in the absence of
to M, subterranea or species of Nannocftlolopsis, if phylogenetic relatedness may most cells will recover a con
also indicate phenotypic similarity. Thus, it would be interesting to test additional depleted cells. Synurophyte
unicellular members of both classes, Eustignatophyceae (e.g yischeria Pascher, Some species are regarded
Pseudostaurastrum Chodat, Eusligmaros l) J Hibberd; l'ig.2.2b, Trr{c,4ydi.scus Ettl) characterisiic of eutrophic 1

and Xanthophyceae (e.g. Chlorellidium Vischer, Misc,hococcus Nägeli), for thcir bio Chrysophytes arc goldel
technological potential. Even in recent literature, there is somc confusion, and nial and occur as flagellat(
certain coccoid yellow-greenish algal species have sometimes been wrongiy interesting morphologies (C

assigned to one of the two classes (e.g. Lang et al. 2011). covering the cell surface ar(
slightl;r acid fteshwaters o
species richness increase w

2.5.3 Other photosynthetic Stramenopites ecological impacts becaus(

metabolize djssolved organ
2.5.3.1 Raphidophyceae thesize (Graham et a].2001
Membcrs of this class of stramcnopiles, also called "chloromonads" are unicellular
tion of undesirable blooms
fatty acids that affect fish r
flagellate algae with about 50 known species that occur in marine, brackish or
can give it an unpleasant t
freshwater habitats. lhe freshwater spccies of the Raphidophyceae are green'
whereas the marinc forms are yellowish and contain the carotenoid fucoxanthin.
The chloroplasts of Raphidophyceae are usually small and numerous per cell. What
2.5.3.3 PhaeoPhYceae
makes Raphidophyceae interesting for biotechnological applications is thai their
photosynthetic assimilation products are oils that are stored as droplets in the This class comPrises the
includes the most conspi
cytosol. Some marine Raphidophyceae, e.g. Chatonella antiqua aud lleterosigmQ
Northern Hemisphere. Mo
carteJ"ae, produce strong neurotoxins and cause toxic red tides. A species of Olistho-
several metres long (e g. I
riiscus N. Carter has been used in biotechnological appllcations because of its
intertidal belt and the uP
rather high content of the fatty acid EPA, and it also produces p-1'3 glucan (Mar
haryested and are of imPo
shall et al. 2002).
and are important edible
algae, they are l]ot treale
(2009).
2.5.3.2 Synurophyceae and Chrysophyceae
Members of both classes of Stramenopiles are close relatives in molecular phyloge-
nies (Yang et al.2012); synurophytes and chrysophytes are able to switch between 2.6 Chromalveolate
autotrophy, heterotrophy and even phagotrophy, which puts them at a distinct algae
advantage in aquatic ecosystems that are low in nutient concentrations and have
rcduced light penetration. This may be advantageous in certain biotechnological Haptophyte algae are a m(
applications as well where the provision of sufficient light and nutrienfs can be isms with a haptonema'
an expensive cost factor. However, neither group has been used in any biotechno between two approximate
logical applications until now. in great abundance, even
Cell surfaces of synurophytes are covered by overlapping sjlica scales, some- share many featu(es, e.g l
times with spiny bristles, and are perforated (Graham et al. 2009) The scales' matic reticulum and Plast
perforation patterns are used as taxononic features to dclimitate species of synuro' the photosYnthetic Stram€
phytes. A number of adhesive polysaccharides are involved in the scale attach' ular evidence, the haptop
 - 2.6 Chromalveolate älgae: coccoLithophorids and haptophyte algae 31
-
tic ment. ln conlrast to most diatoms, synurophytcs can continue to divicle and futtc
AI tion also in the absence of an external silica covering. In cullures of naked cells,
ay most cells will rccover a compiete cell covering when silicate is rcsupplied to silica
al depletcd cells. Synurophytes are abundant in neutral to slighlly acidic fieshu'atcr.
3r, Some spccies are regarded as indicators of low levcls of pollution, but others are
tl) characteristic of eutrophic lal<es (Craham ct al. 2009).
o Chrysophytes are golden brown micro algae that are mostly unicellular or colo-
'rd
nial and occur as tlagcllates or non-motile cells; thcy include many unique and
lly interesting morphologics (Grallam et a].2009). In contr.tst to synurophytes, scales
covering lhe ccll surface are absent in chrysophytes. Chrysophytes iypically favour
slightly acid frcshlvaters of moderate to low productivity; lheir abundancc ancl
species richness increase u/ith lake eutrollhic status. Chrysophytes may have strong
ecological impacts becausc many are mixoirophs thal are able to take up aDd
nletabolizc clissolved organic compounds and Particulate food as well as photosyn'
thesize (Graham et al.2009). Several chtysophytes are associated lvith the torma-
AI tion of undesirablc blooms because living cells of certain species can produce toxic
OI fatty acids that affect fish or excrete aldehydes and ketones into tltc lvater' rvhich
nJ can givc it an unpleasant taste and odour.
n.
at
:ir 2.5.3.3 Phaeophyceae
ile This class (lnlprises the brown algae, an almost entirely marinc group which
1A includes the most conspicuotls seaweeds of coicler u/aters, particularly in the
0 Northern Hemisphere. Most brown algae form large thalli, it'hich can be up lo
its several metres long (e.g. the Laminariales or "kclps"). lhey nostly gtow in thc
lt- intcrtidal belt and the upper liltoral region oI sea coasts. These macloalgae arc
harvested and are of important industrial usc, e.g. for the production of alginates,
and are important eclible scar,veeds. Becausc this review focust's on nlicroscopi,
algae, lhey are not treatcd further llere. For an introduction, see Graham et al.
(2009).

ln
2.6 Chromalveolate algae: coccolithophorids and haptophyte
Lct
algae
\ie
:al Haptophyte algae are a nonophylctic group that includes all pllolosynthetic organ-
b.. isms with a haplonema, which is a microtubule-supported appcnd.rge thai lies
0 lletwecn two approximately equal flagella. Most haptophytes are marinL', utcurring
in grcat abundance, evcn itt greatel water depths (up to 200 m). Haptophyte algae
LC
sharc many fealures, c.g. pigmenl composition, prescnce of a chloroplast endoplas-
ts' matic reticulum and plastitls dcriveil tlom red algal secondary enclosymbiosis with
'o- the photosynthetic Str.rmenopilcs (Andersen 2004). llased on shuctural and molec-
h ulaL evidence, thc haptoplrytcs form a monophyletic lineage, treated as the phylum
t2 2 Phytogeny and systematics of microalgae: An overview -
-
group' cells a rnore rigid, infle:
Haptophyta, which is divided into two classes (Andersen 2004)' The larger
patterns, which are ext
class Coccolithophyceae, compdses the coccolithophorids, which are haptophytes
2004). The success of
WithacellcoatofminelalizedScales(coccoliths)andtwoequalflagellaorlack
unique behavioü patte
flagella altogether. Coccoliths are largely composed of calcium carbonate crystals
Iive phagotrophically (f
in ihe form of calcite. Coccolithophorids remove large quantities of atmospheric
are an important in addition to photoaut
CO, through their photosynthesis and calcification and, therefore,
(McConnaughey et a1 1994)' accounting produce toxins that ale
component of the global carbon cycle
other components of th
for a substantial part of the ocean-floor limestone sediments' Coccolithophodds
light; some function as
contribute at least 25 % of the total annual vertical transport of inorganic carbon
of their nutrition. Maty
to the deep ocean (Rost and Riebesell 2OO4). Coccolithophorids have an excellent
reef-building corals. In c
fossil record, e.g. extensive chalk deposits that were laid down' coccoliths
are
past climate and foraminifera, radiolada
widely used as bioindicators in the oil industry and as indexes of lusks (Hackett et al. 200
ocean chemistry conditions (Young 1994). Haptophltes are ecologically
significant
Haptophyte algae are erable number live hete
in terms of both biotic interactions and biogeochemistry'
contain nutri- is used in commercial a
considered as high-quality foods for zooplankton; several species igo) Javornicky. lt is kr
valuable for the
tionally important PUFAs, which makes them also commercially applications and aquac
produce
produition of fish in aquaculture systems. Other coccolithophorids may
toxins that destroy cell membranes or produce copious amounts of organic
slime

and foam (Graham et al. 2OO9). Many coccolithophodds form blooms in ocean
di-
2.8 Euglenoids (Et
waters and produce large amounts of a volatile sulphur-containing molecule'
methyl suffiäe, that enhances cloud formation and increases acid rain' Another Euglenoids are a divers(
cooling effect on the climate comes from the coccoliths, which readily reflect light' half of which contain a
thereby increasing the reflectance of the ocean's surface So far' only two
species that is bounded by thr€
ations' Isochrysis galbana
of Haptoph ta have been used in biotechnological applic ment. The remainder t
part" ana Pavlova luthei (Droop) J. C. Green (Fig 29) are both unicellular flagel' bacteria or other eukz

lates ftom marine envitonments. While I' galbana is naked, the cell surface
of euglenoids may have a
P. Iutheri is covered with cellulose scales. Both species are rich sources of PUFAS evolution, even though
(EPA, DHA) as well as some sterols. They are often used in aquaculture' e'g' to closely related to the p
feed clams and oYsters. nemids, making up thr
euglenoids are that the'
amylon) in their cytoPl
2.7 Chromalveolate algae: Dinoflagetlates (Dinophyta) posed of parallel dbbo
Leedale and Vickermal
The dinoflagellates are an important group of phytoplankton in marine
and fresh-
attracts interest for bi(
is reflected by a tremen-
waters. The adaptation to a wide vadety of environments tent of paramylon whr
dous diversity in form and nutdtion and an extensive fossil record dating back
cally, it is a rich sourct
cells have tlvo
several hundred million years (Graham et al. 2009)' Dinoflagellate highest concentration.
swim'
structurally distinct flagella whose motion causes the cells to rotate as they
but also non flagellate unicells and filamentous forms of dinophytes are known'
covering
Two different cell types can be distinguished on the basis of the cell-wall Acknowledgemenl
or theca, The "naked" or unarmored forms have an outer plasmalemma surround-
dino- We would like to thantr
ing a single layer of flattened vesicles (membrane sacs or alveoli)' Armoured
gMng the sions and suggestions.
nagellates have cellulose or other polysaccharides within each vesicle'
 Acknowledgements _ 33
up,
cclls a morc rigicl, inflcxibio lvall.
These cellulose plates are auanged
tes patterns, which are extensivol),u_sed in.listinct
as raxononic ::i,"*"r0,,",r-
rck (iiacke ct
2004). the success of alinoflaspllarcs
prryt"pi"rri"ilrlr' ," n"" in parr al.
als
unique bchaviour pirtterns' incruding ", to
ric ai"r'r*ti."i'rö"trontano
';^.;':";i. tteir abiljty to
live phagotrophicat! (f.eecline on p",ri.t",
nt ;:;;,
in addjrion to phoroaurotrophir:aIy (mixotnrphic ;, other organis,,)
1g produce toxins that are ditngel_olls h'r;;,";.';" dinoflagclates
to tnan, marine _"nln ufs' i,rn, seabjrcls,
ls othcr conrponenrs of the marine ancl
food chain. Orh"r. ;;" ;i;i";inescent
)n light; some functioD as parasites anct enit
o_r syntbjonts that rellr
1t of their nurrition. Many photosynthetir:
on ho,st organisms for part
'e
clinofragellates rr" u, .'',,n
reef-building corats. L
other svmrrturt. .unonug"liui" "rioru.no,on,.
I ;";;;, *"
d fotaminifera, raciiolarians, flatrror,ns, hosrs inctucle
anemones, jeliyfish ancl even
lt lushs (fiackett ct al.2004). While bivaive mol-
rrost ciino agcllate.lr"
e erable nu nber live hererotro phjcally. ,,r,,,"*.,"*aic, a consitl
s" f",, ,r;u .,r;i; ,p".io"ror linun og"t
is used in conlnrercial applications, " tut",
hcterotiophit: 7 ,pri".rii,Urn^cohnii
the.
(Scl
Jgo) /avornicky. It is known f.r irs high.on,"nr'oiPuiar,'.ä
applications and in unriuu, toua
aquaculturc, and to procluce
oh"r.";;;;.";-
2.8 Euglenoids (Excavata supergroup)
Errqlcnoids are a djrerse qrouo
ofcommon marjne ancl freshwater
hait of which tontain a pJastid derived flagellates, about
,r"nr;"J;,;, .;;.inniru o,nr,.;
that is bounded by three nrembr
and contains chlorophyll ö as "nnorun
accessory
r.'cnt. The remainder o, ,n" nr.onut pr.g

,,"' ;l:,:';:;::;.:il'J;*,_:' ffj;

",,'
",' ",
eugienoids
i",":;,.;:,
may have acquired Lhejr.plar,i
l;:ir;r j[:, ;:lJ ;:
evolution, e\r,n thoueh rhe cuglenoids
tr'f;'; ;;;";,;;;ffi;;,:," :;
closely telJteLl to thc p,rra5jtic
rrroy rr" o ,"ti"",
uia Jrirp uugrnnoia.
trypanosomes (l(inetoplasicls), "ro
n.mids. together with tliplo
nr.rtirq up t'e rnononhylg,. lugt"nnrou.
cuglettoirls are rhrr rh*r pruclrr, ii;;;;il;,"* fearures oI
*a stoj.rgF(Jlbohvrlld,.. p ,.,.i,ri",f qlrr,.rrn
anyton) in thcir cytopiasm, and th' " r1r.11
they dirpi.; .
pr'.ed crl p;rallcl rihbon likp protFina.eou\ ;;iq;J;o.i r,.,.aur" .o,,'.
srripcj. l-or ,*i*,ir'lf
Leedaie and Vjckeflnan (20oOl
or "r*f"n,,iar, r."
terest r", I"::rlli:I, i;l ilT
:-rlTc:s
in
tent of paramylon when
bi
","J;;i";i,
grolvn hetcrotrophicalli,. Wt
::, ; i,f n:nl:;tH:
g.o;u,, piotoautotlophi
cally, it is a rich sourcc oI the antioxid.rnr
a."on"r"i,"n o..urc in l"
any algae "(]_ani ancl Tsumura
highest concenh.ation l<nown for "irnrl'tr,lir
l9g9).

Ac kn owled ge m en ts
l4le would like to thank
Clenens I,i and carola Griehl' for their helpful
sions and sLlllgestions. *" u.Lno*uttnn discus
ledge thc support available lhrough
cbooks dis
74 l Phylogeny and systematics of microalgae: An overview
-
-
Friedl, T. and M. lorenz.2
tributed by "Alginet", which was a thematic network project funded by the Euro- a biological resource
pean Commission (proiect no. QLK3-CT-2002-02132). Parts of this review were \atenaes 7t. LLU- rLt,
(contract
financially supported by the State of Lower Saxony' Hannover, Germany Friedl,t and N. Rybatka.:
no.2N2727). Cunent Status. ln: Pr
and Dennis trancis, e
Gould, s.B., R. F. Waller at

491-517.
References Graham, l. E., L. w. Wilcol
Beniamin Cummings,
Adl, 5. ivl., A. G. B. Simpson, M. A. Farmer' R. A. Andersen, O R' Anderson' l' R' Barta' S' 5' Guiry, M. D. and G. M. Gu
Bowser, G. Brugerolle, R. A. Fensome, S. Fredericq, T. Y James, S Karpov' P Kugrens' University of lreland,
R M' ivlccourt' L' Mendoza'
,. Krug, C, E. Lane, L. A. Lewis, J. Lodge, D. H. Lynn, D G Mann, Hackett, J. 0., D. lvl. Ander
0. Moestrup, S. E. I
Mozley'standridge, A. Nerad, C' A Shearer, A' V' Smirnov' t' W Spiegel
remarkable evotution
and M. F. R. Taylor. 2oo5' The new higher level classification of eukaryotes with emphasis
J. Hempel, F., A. Bozarth, N.
on the taxonomy of prolisls- J. Eukotyot, Mictobiol' 52' 399-457' Maier.20l1a. Microa
Bot
Andersen, R. A. 2004. Biotogy and systematics of heterokont and haptophyte algae
Äm' '/ 81.
91,7508-7522. Hempel, F., J. Lau, A. Klinl
Andersen, R. A. 2008. Moon Boad. Limnol- Oceonogt Bull' 17' 8-9' Human Antibody and
Archibald, ,. M. 2009. Green Evotution, Green Revotution Sc'etce J24' 191 792' One 6t e28424.
Baurain, 0., H. Brinkmann, l. Petersen, N. Rodriguez-Ezpeleta, A Stechmann' V Demoulin'
A J'
Holzinger, A. 2009. Desic
Roger, G. Burger, B. F. Lang and H. Philippe.2olo. Phytogenomic Evidence for Separate
structural requiremer
Actuisition of Plastids in Cryptophytes, Haptophytes, and Stramenopiles /vlol' Biol' Evol' 27' engergy sources. No\
769a-1709. Huss, V A. R., C. Frank, E
Becker, B. and B. Marin.2009. Streptophyte algae and the origin of embryophytes
Ä'n Bot-
E. Kessler, 1999. Bio
London 103,999-7oo4. sensu lato (ChloroPh
Bhattacharya, D., i. M, Archibald, A. P. M. Weber and A. Reyes-Prieto 2007 How
do
John, D. i\4. 1994. Biodive
endosymbionts become organetles? Understanding early events in ptastid evolution' t
Kauff, and B. Büdel.20
Bi oessoys 2 9, 1239'1246. Progress in Botany 72.ln
Bigogno, C., l. Khozin-Goldberg, D. Adterstein and Z Cohen 2o02a Biosynthesis
of arachidonic Francis, eds), SPring
acid in the oteaginous milJoalga Porietochloris inciso (Chlorophy'eae), Radiolabeling Keeting, P J.2004. Diver!
studies. Lipids 37, 209-276. L487-1493.
Bigogno, c., l. Khozin-Goldberg, S. Boussiba, A. Vonshak and z cohen 2002b Lipid
and fatty Keeling, P J.2010. The e
grein oleaginous alga Porielochloris inciso, the richest plant source Tronsactions of the I
acid composition of the
of arachidonic acid. Phytochehistty 60, 497-5o3. Keeling, P,., G. Burger, I
(B Bolin'
Bolin. 8., E. T. Degens, P. Duvigneaud and 5. Kempe. 1977 The Global Carbon Cycte' /a: M. W. Gray. 2005. Tt
T. Degens, S. Kempe and P Ketner, eds) The GlobalCarbon Cycte Wiley' New York' pp 1 Keeling, P.l. and I, D. Pa
E.
53. Genet, 9,605-618.
Bozarth, A., U.-G. Maier and S Zauner. 2009. Diatoms in biotechnologyr modern
tools and Krienitz, 1., E. Hegewald,
apptications. /ppl. Mictobiol. Biotechnol. 82, 795-201' relationship of Chlor
the Trce Phycologia 43,529-
Brodie, J. and G. Zuc.arello. 2006. Systematics of the Species Rich Atgae ' Reconstructing
of Life. CRC Press. PP. 323-336. Kroth, P. 2007. Molecular

Büdet, 8,,f Darienko, K. Deutschewitz, S. Doiani, T. Friedt, K' Mohr, M Satisch' w- Reisser and Microalqae as Grcet
Springer New York
B,Weber'2oog.southernAfricanbiologicalsoilcrustsareUbiquitousandhighlydiversein
Lang, 1., L. Hodac, T. Frie
drylands, being restricted by rainfatl frequency Mictob Ecol 5T' 229'247'
patterns in microalg
Cohen,2.7999. Chemicols fron nicrcolgoe. Taylor & Francis, London'
of cutture cotlection. S
Csögör,2., D. Melgar, K. Schmidt and C. Posten. 1999' Production and particte characterization
the frustules of Cyclote llo cryptica in comparison with siliceous earth' /' Biotechnol' 70' 77-
Lebeau, I and l.'M. Robr

Part li Cultivation at
75.
Edition' Lebeau, T, L. and l. Nl. R
Friedl, T. and B. Büdel. 2008. Photobionts. ,r: (T. lll Nash, ed ) Lichen Biology 2nd
products. Part ll: C!
Cambridge University Press, cambridge. pp. 9-26.
--
References 35
-
Friedl. T. and tu1. Lorenz. 2012. The Culture collection of Atgae at Göttingen Llniversity (SAG)l
a biological resource for biofechnological and biodiversity teseatch Procedio Environmental
sricn.es 15. 71O 777.
Friedl, T. ancl N.Rybalka.2012 Systematics ofthe Green Algae: A Brief lntroduction lo the
Current Status. ht Progress in Botany /3 (Ulrich Lüttge. Wolfram Beyschlag, Brrr(hard Büdet
and Dernis FIarcis. eds). Springer Berlin Heidelberg. pp 259-280
Gou1d. S.8., R. F. Waller and G. L I!4cFadden 20O8. Plastid Evolulion. Än,u. Rev Plont
Biol 59,
491 517.
Graham, J. E., L. W. WiLcox and L. E. Graham. 2OO9. Algae (2nd Edition).01 Dec 2008 Edilion-
Benjamin Cumm jngs, 720.
Guiry, Nl. D. and G. t\4. Guiry.2012. AlgaeBase World_wide electronic publication, NTiional
University of lreland, Galway.
Hackett, l. D., D. J\'1. Anderson, D. L. Erdner and D Bhattacharya. 2004 Difoilagetlales: A
remarkable evolutionary experiment Äm. gat 91"1523 1534.
/
Hernpe[, F., A. Bozarth. N. Lindenkamp, A. Ktingl, S. Zauner, U. Linne' A. SteinbucheL and U'
j!4aier. 2011a. I\'licroalgae as bioreactors for bioplastic production. l\,11icrobj01 CPll Fo'totics l0
81.
Hempel, F., I. Lau, A. Klingt and U- G. l\laier. 2011b. Algae as Protein Factories: Expression of a
Human Antibody and the Respectlve Antigen in the Dlatom Phoeodoctylum tri'arnutun Plos
One 6, e2a424.
HoLzinger. A. 2009. Desiccation toterant gleen algaer inrplications of physiological adaptation and
structuraI requirements. /rr (Krislian N Hagen, ed.) ,4/9oe: ,'r utrition' pollution 'ontrol and
engergy saurces. Nova Science, New York. pp 41 56.
Huss, V A. R., C. Frank. E. C. Hartmann, /\,t. Hirmer, A Kloboucek, B. M. Seidel, P Wenzeler and
E. Kessler. 1999. Biochemical taxonomy and rnolecular phylogeny of the genus Chlarello
sersu lalo (Chlorophyta). J. Phycal. 35,587 598
lohn, D. Nl. 1994. Biodiverslty and Conselvation: An Algal Perspectlve The Phycoloqist, 38,3
15'

Kauff. F. and B. Büdel.2011. Phvlogery of Cyanobacteriai An Overview

Progress in Botany 72. /t: (U{rich E. Lüttge, Wolfram Beyschlag, Burl(hard Btidel and Dennis
Francis, eds). Springer Berlin Heidelberg pp.2o9 224
/
l(eeling, P. J. 2004. Diversity and evolutionary history of plastids and their hosts Ä'r' 80t 91'
14a1 1497-
Keellng, P. J.2010. The endosymbiotic origin, diversificatior and fate ol plastids. Philosaphical
Tronsactions ofthe Royal Society B: EioLogical Sciences365'729 748
Keeling, P J., G. Burger, D. G. Durnford, B. F Lang, R. W. Lee, R. E. Pearlman, A l
Roger ard
l\il. W. Gray. 2005. The treeof eukaryotes. frends Ecol. Eval 2A,670-676
K€eling, A J. and J. D. Palnrer. 2008. Horizontal gene transfer in eukaryotic evolution /Vdt Rev
Genet. 9. 605 618.
Krienitz, 1., E. Hegewald, D. Hepperle, V A H. Huss' T. Rohr and M. Wolf 2004 Phylogenetic
relaiionship af Chloretlo and Parochlarella gen nov (Chlorophyta, TrebouxiophyceaeJ
Phycolaqia 43,529 542.
Kroth. P. 2007. Molecular BloLogV and the Biotechnologlcal Potential of Diatoms. ln: Tronsgenic
Microalgae as 6reen Cell Factoies. (Rosa Le6n, Aurora Galvän and Emitio Fernändez, eds)'
Springer New York. pp. 23-13.
Lang, 1., L. Hodac, T. Friedl and l. Feussner' 2011. Fatty acid proflles and their distribution
patterns in microalgae: a comprehensive analysis of more than 2OO0 strains frorn the SAG
culture collection. BMC Plont Biol. 11,724
Lebcau, T. and J. M. Robert. 2003a. Diatom cultivation and biotechnotogicälly relevanl products
Part l: Cultivation at various scales. Äppl. Alicrobiol. Biotechnol. 6A,612-623
Lebeau, T. L. and I. tV\. R. Robert.2O03b. Diatom cuLtivation and biotechnologically relevant
products. Part ll: Current and putative products Äpp1. Microbial Biotechnol 60,624-632'
t6 2 Phylogeny and systematics of microalgae: An overview
-
pp 560 pp' Pröschold, T. and F. Leliaen
Lee, R. E. 2oo8. Phycology (4th edition). Cambridge lJniversity Press, Cambridge:
t approaches. /t: 0 Bro(
Leedale, G. F. and K. Vickerman. 2o0O. Phytum Euglenozoa /n: (J lLee, G' Leedate and P'
future of algol systemt
Bradbury, eds) A, illustrated guide to the Wotozoa, 2nd ed Society of Protozootogists'
Pulz, O. and W Gross.200,
Lawrence. Kansas, USA. pp. 1135-1185.
Leliaert, F., D. R. Smith, H. Moreau, M. Herron, H. Verbruggen, C F Delwiche and
o De Clerck' Microbiol. Biote.hnol.
green algae' Ctit' Rev' Plant Sci 31'7-46 Rindi, F. 2011. Terrestriol 9t
2012. Phytogeny and molecular evolution of the
as Gteen Cell Foctories climote chonge.ln Clit
Leön, R., A. Galvän and E. Fernändez. 2ooT Tronsgenic Mictoalgae
Michael B. Jones, Step
Springer New York.
in Press.
Lewis, L. A. 2007. Chtorophyta on land. /r: (,oseph Seckbach' ed\ Algae and cyanobocteria
Rodriguez-Ezpeleta, N., H.
extrcne e nvi rcn m ents. Springer, Dordrecht. pp, 569-582'
soil green Bohnert, H. Philippe a
Lewis, L. A. and B A. Lewis. 2005. unearthing the molecular phylodiversity of desert
Green Plants, Red Algi
algae (Chlorophyta). syst. Biol 54' 936-947.
/
plants Am Bof 91' Rost, B. and U. Riebesell. i
Lewis, L. A. and R. M. Mccourt. 2004. Green atgae and the origin of land
to environmental chan
1535-1556.
Moleculat Processess
L6pez"Bautista, J. M., F. Rindi and D. casamatta. 2007' The systematics of subaerial atgae'
Sharma, N., S. Tiwari, K. Tr
/n: (ioseph Seckbach, ed.) Algae ond cyanobocte o in extreme envircnments' Sptinget'
algae): facts and chatl
Dordrecht, pp. 601-617.
Tani, Y and H. Tsumura. 19
Luo, W., t Pröschold, C. Bock and L. Krienitz. 2olo Generic concept in Chlorelta'related coccoid
tocopheroI production
green algae (Chtorophyta, Trebouxiophyceae). Plont Biology 12' 545-553'
Tanoi, T., lvl. Kawachi and,
lvlarshall,,.-A., P D. Nichols and G. lvl. Hatlegraeff. 2OO2 Chemotaxonomic survey of sterols and morphology of Eotryo,
fatty acids in six marine raphidophyle algae. J. Appl' Phycol 14' 255-265'
in the Tirichine, L. and C. Bowler.
Martin, J. H., G. A. Knauer, D. M. Karl and W. W Broenkow lgSZ VERTEX: carbon cycling photosynthetic life on
northeast Pacific. Deep Sea Reseorch Part A. Oceanogrophic Research Papets 34' 267-285'
photosynthesis' and Ward, D. and R. Castenholz
Mcconnaughey, I A., D. traneois, A. Denis and T. Anne. lgg4 Catcification, Cyanobacteria. /nr (Bri
global carbon .ycles. Bulletin de l'lnstitut oceonographique (Monoco)t 737-767'
Weiss, T.1., J. Spencer Johr
MedLin,L.andl.Kaczmarska.2004.EVolutionofthediatoms:VMolphologicalandcytological Devarenne.2010. Phy
support for the major clades and a taxonomic rcvision' Phycologia 43,245-270' hydrocarbon-producin
lvlilanowski,R.,s.Kosmata,B.zakrySandJKwiatowski'2006Phylogenyofphotosynthetic (Chlorophyta). /. P/,yc(
euglenophytes based on combined chloroplast and cytoplasmic SSU rDNA sequence
Wen, Z.-Y and E Chen.20(
analysis. J. Phvcol. 42,721-730.
Biotechnol. Adv. 21,2
lvloustafa,A.,B.Beszteri,U.G.Maier,c.Bowler,K.ValentinandD'Bhattacharya-2009Genomic Wodniok, 5., H. Brinkmann
Footprints of a Cryptic Ptästid Endosymbiosis in Diatoms S'ietce 324' 7724 1726' 2011. origin of land p
lvlüller, 1., T. Friedl. D. Hepperle, lil. Lorenz and l. G. Day. 2OO5 Distinction between mutliple Worden. A.2.. l. K. Nolan a
isolales of chlorclla vulgdrrs (Chlorophyta, Trebouxiophyceae) and testing for conspecificity Picophvtoplankton: Tl
using Amplified Fragment Length Polyinorphism and ITS rDNA sequences' J Phycol'
41'
76A 779.
7236-1247. Yang, E. C., G. H. Boo, H.l
Nakada,l,K.MisawaandH.Nozaki.2o08.MolecularsystematicsofVolvocales(chlorophyceae, Supermatrix Data Higl
chlorophyta) based on exhaustive 18S rRNA phytogenetic analyses Moi Phylogenet' Evol' Protist 163,277-237.
48,2A1-297. Young, I. R. 1994. The fun{
Norton,lA.,lvl.MelkonianandR.AAnd€rsen.1996AtgalbiodiversityPhycologia3S'30a-326 coccolithophorcs. Cat
proximate biochemical
Pahl, S., D. Lewis, F. Chen and K. King. 2010. Growth dynamics and the
composition and fatty acid profite of the heterotrophically grown diarorl. Cyclotello cryptico'
l. Appl. Phvcol. 22,165-777.
Perez-Garcia, O., F.M.E. Escalante, L. E. de_Bashan and Y Bashan 2011' Heterotrophic cultures of
microalgae: Metabolism and potential prcducls' Watet Res 45,71-36
Piibyl, P, M. Eliä!, V Cepäk, J. Lukavski and P. Kaitänek' 2012' Zoosporogenesis' motphologv'
ultrastructure, pigment composition, and phylogenetic posilion ol Trachydis'us minutus
(Eustigmatophyceae, Heterokontophyta) 1. /. Phycol 4S' 231-242'
Pröschotd, T., T. Darienko, P. C. Silva, W. Reisser and L Krienitz' 2011 The systematics
of
ZoochLorella revisited employing an integrative approach Envircn' Mictobiol' 13'350-364'
---

Relerences 37
-
Pröschold, T. and F. Leliaert. 2007. Systematlcs oithe green algae: conflict of classic afd modern
approaches. /r: (l Brodie and I Lewis, eds) Unravelling the alqoe: the past, present, and
future of algal syslenati.s. CRC Press, Taylor & Francis, Eoca Raton. pp. 123 153.
Pulz, O. and W. Gross. 2004. Valuable prodLrcts from biotechnology ot microalgae. App1.
l\4icrabiol. Biotechnol. 65, 635 64A.
Rindi, F. 2011. ferrestrial green algae: systemotics, biogeagraphy and expected respanses to
climote change.lr Cllmate change, ecology and svstematlcs, eds. Trevor R. Hodkifson.
IVllchael B. lones, Stephen Waldren and lohn A. N. Parnell. Canibridge: Cambridgc UniversitV

Rodriguez-Ezpeleta, N.. H. Brinkmann, S- C. Burey, B. Roure, c. Burger, W Lötfelhardt, H. J.

Bohnert, H. Philippe and B. F. Larg. 2005. l\4ofophyly of Primary photosynthetic Eukaryotes:
Green Plants, Red Algae, and claucophytes. Cuft. Biol. 15,1i25-133O-
Rost, B. and U. Riebesell.2004. Coccolithophore calcification and the biological pump: Response
to environmental changes. /n: (H.R. Thierstein and J. R. Young, eds) totrclithophares - Fram
Moleculor Processess ta Glabal lnpact. Springer Verlag, New York. pp.99-126.
Sharma, N., S. Tiwari, K. Tripathi and A. Rai.2011. Sustainability and cyanobäcteria (blue,green
aLgae): facts and challenges. /. Appl. Phytol.2J. 1059-1081.
Tani, Y and H. Tsumura. 1989. Screening for tocopherol prodLrcing microorganisrns and alpha-
tocopherol production by Euglena gratilis. Z. Agricultural ond Biol- Chen. 53, 3OS 312.
Tanoi.l, [4. Kawachi ärd 1\4. Watanabe. 2011. Effects of carbon source on growth and
morphology of Botryococcus braunii. I. Appl. Phycol. 23,25 33.
Tlrichine, L. and C. Bowler. 2011. Decoding algal genomes: tracing back the history of
photosynthetic life on Earth. The Plant J. 66. 45 57.
Ward, D. afd R. Castenholz- 2002. Cyanobacteria in Geothermal Habitats. The Ecolosv of
Cyanobacteria. /r: (Brlan Whitton and Malcolm Potts, eds). Springer Netherlands. pp. 37 59.
Weiss, T.1., J. Spencer lohnsion, K. Fuiisawa, K. Sumimoto, S. O (ada, J. Chappctt andI P
Devarenne. 2010. Phylogenetic placement, genorne size, and GC content of the liquid.
hydrocarbof-producing green microalga Botryacoccus bnunii a.Üalr Ber(eley (Showa)
(Chtorophyta). /. Phycal. 46,5i4 54A.
Wen, Z. Y and E Chen. 2003. Heterotrophic production of eicosapentaenoic acid by microalgae.
Biotechnol. Adv. 21, )73 294.
Wodniol(, S-, H. Brinkmann, c. cloc (ner, A. Heidel. H. Philippe, M. Melkonian and B. Becker.
2011. Origin ol lard plaft5: Do conjugating green algae hold the key? Bnc Evol Bial 11,1A4.
Worden, A-2., l. K. Notan and B. Palenik. 2004. Assessing the Dynamics ard Ecology of Marine
Picophytoplan(ton: The lnrportance of the Eukaryotic Component. Linnol. Oceonogr.49,
164 1/9.
Yang, E. C., G. H. Boo, H. l. Kim. S. M. Cho, S. [4. Boo, R. A. Ar]dersen and H. S. Yoon. 2012.
Supernratrix Data Highlight the Phylogenetic Relationships of Photosynthetic Stramenoplles.
Protist 163,217 231.
Young, j. R. 1994. The iunctions ol coccoLiths. ir: (A. Winter and W. c. Siesser, eds)
Coccolithophores. Cambridge lJniversity Pr€ss, Cambridge pp. 6l 82.