*

6 CHLOROCOCCALES REPRODUCTION 27

Sexual Reproduction. (Figs. XII-XIII) In Chlorococcum, the swarmers some-

times behave as gametes. The same phenomenon takes place also in Chlorochytrium
lemnae, C. limnanthemum and Rhodochytrium. The functioning of swarmers either as
.t zoospores or gametes is considered rather primitive (Fritsch, 1935).
The fusing gametes may be of equal size as in Rhodochytriumor they may be of
a equal or unequal size as in Chlorococcumand ChlorlJchytrium. The zygote formed by the
fusion of the gametes develops into a new individual. Fusion between similar or
dissimilar gametes has also been knowl;l in Trebouxia when the gonidia taken from the

~j .
lichen body of Parmelia are grown in pure. cultures. In Characiositphon,the biciliate
gametes are isogamous or anisogamous (Iyengar, 1936, 1954), theptant being diQe"

f
cious. Isogamy may take place between larger gametes as well as between smal.ler
gametes. In Pediastrum and Hydrodictyon, gametes are produced in large numbers i~
the parent cell and they are smaller than the zoospores and isogamous. Unlike the
g zoospoJes, they are liberated individually through a hole in the parent cell membrane.
Hydrodictyonis monoecious and even gametes from the same coenocyte may copulate.
The zygospores on germination give rise to four swarmers which develop into
polyhedral cells and these may be easily mistaken for Tetraedron cells (Fritsch,
. - 193?, Fig. 50, I, K). These polyhedral cells give rise to swarmers whith combine to

~. 7~
form a new net.
Korshikoviellalimnetica (=Characium limneticum)'...d Rhopalosolensaccatus(=Characium
0, k 8m saccatum)produc~ gametes which are distinctly anisogamous. In the latter, individuals
which are prpbably purely sexual, produce macro- and microgame_tes. In Phyllobium,
the branched coenocytic threads swell up at places to form elongate or globose
resting cells or gametangia. These enlarge due to accumulation of the p.rotoplasmic
contents, and, on the leaves of the host th~y' appear as bright green nodal swellings
(Fritsch, 1935). The gametangia produce a limited number of macrogametes or
a large number of microgametes, both being biciliate. Sexual fusion takes place.
between a micro- and macrogamete, the latter engulfing the former completely with
n
o p
the result that the zygote:shmys only two cilia. .
Oogamous sexu~l reproduction of a fairly high order is known in some members
of-the Micractiniaceae and the Dictyosphaeriaceae._ Ii: was first reported in
FIG. XIII, SEXUAL REPRODUCTION (OOGAMY)IN THE MICRACTINIACEAE,
Golenkiniopsis longispina (=Golenkinia lqngisPina), G. solitaria (=Golenkinia solitaria)
OOCYSTACEAE, AND DICTYOSPHAERIACEAE -and Micractinium pusillum oy Korsliikov in .1~37, and then in Dictyosphaeriumindicum
a-e, Golenkilliopsis minutissima (IYENG. ET BALAKR.) COMB. NOV.; a, MALE
(Iyengar and Ramanathan, 1940). Recently, it has also been reported in Golenkini...
CELL AFTER LIBERATION OF AN ANTHEROZOID; b, FUSION OF THE ANTHERCZCID WITH opsis minutissima (=Golenkinia minutissima Iyengar et Balakrishnan) and in Oocystaenium
THE EGG CELL; C, ZYGOTE ESCAPING OCT OF TilE FEMALE CELL; d-e, ZYGOTES
ATTACHED TO THE OOGONIAL WALL; d, WITH SMOOTH MEMBRANE; e, WITH A SPINY
Gonzalves et Mehra (Oocystaceae). In these genera, the gametangium is an unmodi-
WALL; f-m, Dictyosphcerium indicum IYENG. ET RAMANAT., f, PORTION OF A MALE fied vegetative cell. In Golenkiniopsisand Micractiniumthe single egg formed from each
COLONY WITH FULLY FORMED ANTHERIDIAL CELLS; g, DISCHARGE OF THE ANTHERC- oogonium is retained within the oogonium at the time of fertilization. The anthero-
ZOIDS FROM AN ANTHERIDIAL CELL; h, PORTION OF A FEMALE COLONY SHOWING PAIRS
OF DISCHARGED EGGS; i, TWO EGGS JUST DISCHARGED FROM THE MOTHER CELL (NOTE, zoid, which is probably formed singly from each male cell, as in G. minutissima or in
CONTRACTILE VACUOLES); j, ANTHEROZOIDS SWARMING ROUND AN EGG (PY, PYRE- hirger numbers as in the other species of Golenkiniopsisand Micractinium, is naked or.
NOID; n, NUCLEUS); k, ANTHEROZOID ABOUT TO FUSE WITH AN EGG; I, JUST AFTER
FUSION OF ANTHEROZOID WITH EGG; m, a FULLY FORMED ZYGOTE; n, q; Oocystaenium possesses a cell wall which is discarded just before fertilization. The oospores in.
elegans GONZALVES ET MEHRA; n-o, FORMATION AND UBERATION OF ANTHERO-
OIDS; p, OOSPHERE SURROUNDED BY ANTHEROZOIDS; q, MATURE ZYGOTE.
Golenkiniopsislongispina and G. minutissima are also provided with a firm spinous wall,
as in Micractinium pusillum. In DictyosPhaeriumindicum, which is dioecious, each cell
(a-e, AFTER IYENGAR & BALAKRISHNAN,1956 (AS Go/enkinia minutusima of a female colony produces two eggs which are discharged outside and the cells of male
IYENG. ET BALAKR.); f-m, AFTER IYENGAR ~ RAM"'NATHAN, 1940; n-q, AFTER
GON?;ALVES~ MEHRA, 1959). colonies give rise to 16-32 naked antherozoids, which swarm round the egg. The
 28
CHLOROCOCCALES
zygote is smooth-walled. In Oocystaenium,the male cells produce 16-32 antherozoids
and the female cells a single large egg which protrudes through an opening formed by
the rupture of the cell wall of the female cell. The zygote develops a fairly thick
cell wall, the middle layer of which is verrucose. It is interesting that in the four genera
with oogamous reproduction referred to above, asexual reproduction is azoosporic,
DictyosPhaeriumterrestrebeing the only exception.
CYTOLOGY AND LIFE-HISTORY
The Chlorococcales are characterized by the lack of vegetative cell division, a
feature shared by another order of the Chlorophyceae, the Siphonales. However,
there is one family, the Chlorosphaeraceae (Fritsch; 1935), consisting of some imper-
fectly known algae, which resembles the Chlorococcales in all respects except for the
vegetative cell division. Fritsch (op. c.) appended it to the Chlorococcales, Lemmermann
(1915) included the genera concerned under the Tetrasporales and Herndon (1958)
created a new order, the Chlorosphaeraies, for-the same.
Formerly it was believed (Fritsch, 1935) that all the chromatin substance in the
nucleus of the Chloropbyceae was restricted to the nucleolus (caryosome nucleus)
with little or no chromatin substance-in the outer nucleus. Now, it has been established
that the chromatin present in the outer nucleus during the interphases occurs in a
masked form which does not take up stains easily. In fact, a resting nucleus with a
definite nuclear reticulum is found_ only very rarely in green algae, including the
Chlorococcales. ' ,11'
Even though there is no vegetative reproduction in the Chlorococcales, the
division of the single nucleus takes place before reproduction. Cytokinesis of uni-
nucleate cells is always preceded by a ~itotic division of the nucleus (G. M.' Smith,
1950), Details of nuclear division are more or less as in other green algae. The
, chroIIlOsomes are usually short roas or merely small granules and not very large iIi
number, but in Eremosphaera (Fritsch, 1935), the chromosomes are long and looped.
Cytokinesis in .the Chlorococcales, as in other green algaetis usually 'not by the-
"
formation of a cell plate, but by_the furrowing ofthe plasma..membrane mid-way between
the cell ends. This linear furrow deepens until it cuts through the cell and divides the .
protoplast into two. However, in Eremosphaera cytokinesisprobably takes place by the"
formation of a cell plate. " ~
Two main types' of division 'of tlie~protopl~t to form neW individuals have been
,
recognized in the Chlorococcales (Geitler, 1924; also Fritsch, 1935). In the first type
(e.g., Tetraedron,Characium,Hydrodictyon, Pediastrum,Coelastrum,and probably Scmedesmus),
the number of nuclei in each cell increases without accompanying division of the pro-
toplast so that the mature cell is multinucleate. This is followed by ~'simultaneous
division" of the protoplast by "progressive cleavage" so that a number of uninucleate
protoplasts _are formed (Fig. XIV). These protoplasts, which are angular at first,
get rounded to' form reproductory units. In this type of division the pyrenoid does
not divide, but persists in one of the daughter pro top lasts, only to disintegrate and
disappear sooner or later, new pyrenoids being formed by the daughter cells. In the
second type of division (e.g., Chlorochytrium,Trebouxia, Chlorella,Dictyosphaerium, etc.)
the nucleus divides "successively" and ~.ach division of the nucleus is accompanied.
CYTOLOGY AND LIFE-HISTORY 29
by a division of the protoplast. The pyrenoid also divides during each division of the
protoplast so that each daughter cell receives a pyre.noid. This type of division takes
place in Characiosiphol1also, where the mature cell has a number of separate proto-
, plasmic units, each with a nucleus and a pyrenoid (Fig. IV, f).
However, Fritsch (op.c.) stated that undue emphasis could not be laid on these two
-' types of d.ivision since in some genera both the types ?f division. migh t take \lace. Th~s,
~ Characmm and Sorastrum, zoospores are formed eIther by sImultaneous pr successive
. division of the protoplast. In Sorastrum, division of the nucleus in uninucleate cells is
followed by cytokinesis into uninucleate protoplasts. This is followed by a simultaneous
division of the two 'protoplasts and these in their turn may divide two or three times in
succession(G. M. Smith, 1950). In Chlorococcum,where successive division had been
considered the general rule, some species (e.g., C. multinucleatum Starr) exhibit simul-
taneous division of the protoplast by progressive cleavage of the adult multinucleate
cell (Fig. XIV, a-f).
. In the zoospore formation of Hydrodictyon, blepharoplast granules havo been
observed (G. M. Smith, op.c.).
~. From studies on the pyren91d of Hydrodictyon, Timberlake. (1902) concluded that
the core of the pyrenoid becomes differentiated into two parts, the outer part becoming
a starch layer ultimately. According to this theory, cycle after cycle of starch grains
~y be formed from the pyrenoid (Fig. XIV w). He disagreed with Klebs that starch
can be of two..kinds, viz., pyrenoid starch and stroma starch. According to B~ld
(G. M. Smith, 1950), a somewhat similar process may take place in Chlorococcum. also,
.. ..al!hough he ~ubted genetic continui'ty between-pyrenoid and starch. (see also Bold,
1951). Though a number of workers have supported' TimbeTlake's theory, Fritsch
(1935) held that it was not easy to conceive of the transformation of the protein matter
t.. of the pyrenoid into starch. Further, it would be difficult to explain the production of
starch in some' green algae without pyrenoids. On the whole, he preferred to c::>IIsider
the starch sheaths as being formed from outside.
Electron microscopidal studies (Desikachary, 1959) have revealed photosynthetic
p!'od\!.cts between. the lamellae of the chloroplasts in non-pyrenoid' algae, thereby
confirming Klebs's (1891) findings which distinguish between pyrenoid starch and
stroma starch~ Leyon (1954) also showed presence of chlorophyll in pyrenoids; and
. ,that pyrenoids are permanent organelles which produce starch intensely but still retain
their identity. ' ,
-, As in most Chlorophyceae, the majority of the Chloro<;occaies are haploid, the
diploid condition being confined to the zygote only (Fig. XV). During germination,
the zygote undergoes two successive divisions (Fig. XV d), the first being a reduction
division, and the resulting fOUI cells develop into zoospores or aplanospores. The
meiosis is ~'zygotic" or "initial" (Bold, 1951). However, in a few Chlorococcales like
Chlorochytriumlemnae(Fig. XVI), Apiococcusand probably Phyllobium(Fritsch, 1935)
the first two divisions at the time of swarmer formation in the adult individual are.
meiotic so that the longest stage in their life-histories is a diploid one. In this respect,
Chlorochytriumlemnaeand Apiococcus resemble the Siphonales and the Bacillariophyceae
(Fritsch, 1935). In these instances the zygote germinates directly and the meiosis is
"gametic" (Bold, 1951).
 .~
30 CHLOROCOCCALES ECOLOGY A:-iD PHYSIOLOGY 31

ECOLOGY A1~D PHYSIOLOGY

..
..:.... ,):;.; _.
,:.:--..
,-,' n -;:;..

itnj-.
"

. The majority of the Chlorococcales are free living and planktonic, mostly in shal-
:.:~:::,\:;
~" /~~\
:~}./}..:::: :~:~,)~l.,:.
p

a b
" ...
low confined waters. Some are attached, endophytic, endozoic, parasitic or symbiotic,
whereas a few are found in moist soil or on tree trunks. A few species are also found in
c brackish water or the sea. Yet others form the constituents of snow floras. Some
species can also ~dapt themselves to the high temperatures obtaining in' hot springs.
Quite a number of species are also easily grown in cultures.

"
.: :.:..'.::..
. '"
1.1, J
Planktonic forms. A large number of the Chlorococcales, both unicellular
and colonial, are planktonic, occurring mostly in ponds,. tanks, swamps, small lakes,
.:~
.;,':.' ';':':::1\'"
.:". . ~&..; reservoirs and in rivers with standing water; typical examples being the unicellular
::'::.. ::. :;~".;:..:.:
"-"
";', ~~
':.":' .-.....
," ".:: : . ':':',::: genera, like Schroederia,Golenkinia, Tetraedron, Treubaria and Chodatella, and the colonial
".;::: .:::'; -;:.::::'
genera, Micractin£um, Planktosphaeria,most species of Oocystis and Nephrocytium, most of
'. g. the Selenastraceae, the Dictyosphaeriaceae, the Hydrodictyoideae; the Coelastraceae,
the Scenedesmaceae, the Botryococcaceae and the Radiococcaceae. Most of these forms
are adapted for a planktonic or pelagic habit (Fig. XVII, XVIII). Ih Oocystisand
D
n n II. . . .. .
o
~'-
)...

p
:..::-.:..:.<.:.:<.

;(~fi:;~}'
:..::.;.
.. '.' q
.?~

'.
Nephrocytium the gelatinized parent memorane, and in - Radiococcus, -Kirchneriella,
Gloeoactinium;DictyosPhaeriumand Tetrallal!tos the outer mucilaginous envelopes give
the necessary buoyancy to facilitate a pelagic habit. The flat disc-like or quadrate
colonies of Pediastrum, Scenedesmusand Crucigenia, the irregular colonies of Dimorpho-
coccus, Westella, Selenastrum and Ankistrodesmus, the star-shaped colonies of Adinastrum ,..

~ <~ and the hollow net-like colonies of Coelastrum proboscideumall appear to serve the
. g.;~~1!-~ same purpose. ~n Pediastrum-and Scenedesmusgelatinized bristles are occasion,!Jly found
(Petersen, 1912, '1921) at the apices of the cells. These auginent the capacity for
. '~;:'r:~~
floating. Some like Golenkinia, Micractinium, Polyedriopsis, Treubaria, Chodatella,
Francezaand some species of Tetraedronand Scenedesmushave long spines, bristles or setae.
Sometimes, as in some species of Treubaria, the setae are even gelatinous.
8:+
It is generally known that" the important genera of the Chlorococcales usually
occur during the Warm months of the year, though high summer temperatures are as
~
v z unsuital.?le as l.ower ones" {Fritsch and Rich, 1913), and th~t they constitute one o~ the
important components of the microflora of warm tropical waters. However, h~rdly any
published data is available on the ecology of the Indian planktonic forms of Chlorococ-
~
4 cales, 1)le only accounts available being in connection with taxonomical and morpho-
. logical.studies.. During 1937-40 the author (1940) studied. the ecology of the 'algae of
a pond at Madras, a number of which belonged to the Chlorococcales. Subsequently,
d I
a number of shallow waters in north-east and south India were studied for short or long
bl C I e I
aI

FIG. XIV. INCREASE IN NUMBER OF NUCLEI, CELL DIVISION, AND -+ MULTINUCLEATE PROTOPLAST; 0, YOUNG AUTOSPORES WITHiN THE PARENT CELL;P-U, Pedias/rum boryanum
STRUCTURE OF PYRENOID (TURP.) MENE'GH.; p, YOUNG CELL \\lTH A SIKGLE Nt:CLEUS; q-s; COEKCCYTES \\lTH 2-16 NUCLEI;
t, MULTINUCLEATE PROT<1I'LASTS FORMED BY FIRST CLEAVAGE FURRC.WS; U, COMPLE.TION OF CLEAVAGE
~ a-f, Chlorococcum multinucleatum STARR; a, YOUNG'. VEG. CELi. MEDIAN
OPTICAL SECTION; b-d, CELLS OF VARIOUS SIZES SHOWING INCREASE IN"NUMBER OF
INTO UNINUCLEATE PROTOPLASTS.
ROUNDING UP. NOTE THE
NOTE THE SINGLE DISINTEGRATING PYRENOID; v, PROTOP4STS
NUCLEUS AND NEW PYRENOID IN EACH PROTOPLASMIC BIT; y-d, Chlorococcum
NUCLEI; e, CELL JUST BEFORE CLEAVAGE OF PRC.TOPLAST; f, q.EAVAGE; echinoz.ygo/um STARR, STAGES IN GERMINATION OF THE ZYGOTE FROM STAINED PREPARATIONS, SHOWIKG
g, Spongiochloris excentrica STARR, COMPLETION CF CLEAVAGE BEFCRE ZOOSPOROGE- NUCLEAR DETAILS; W-X, OPTICAL SECTIONS OF TWO PYRENOIDS WITH ASSOCIATED STARCH GRAINS
NESIS; h-I, Scenedesmus quadricauda (TURP.) BRE'B.; h-k, FOUR SUCCESSIVE S.TAGES IN (s, STARCH; P, PYRENOID) IN Hydrodictyon re/icula/um (I..) LAGERH.
THE FORMATION OF A DAUGHTER COENOBIUM; 1, MATURE COENOBIUld; p, PYRENOID;
n, NUCLEUS; m-o, Tetraedron minimum (~. BR.) HANsG.; m-n, CLEAVAGE OP THE ~ (a-g FROM STARR, 1955; h-1, m-o, AFTER G. M. SMITH; p-v, FROM G. M. SMITH,; y-ei,
AFTER STARR, 1955; w-x, FROM TIMBERLAKE).
 £COLOGY AND PHYSIOLOGY 33
32 CHLOItOCOCCAL'£S

M
--'" EI
/i8
I

I¥
0'-:t. "
... ", "
G I '\,
/\.
W..'
i'g';' b
--
A
N'
<+:.:'-''' C
.II d S
I
e

?() a M

-
,
V
,
;
(

I ,
I
.... :

.,'-,. _ 01 .. . .;..

-e
. : t. .
g
I.
@-
I
I
I .8 \- - I rn
!
:(Y

,"
I
I

b
s
Y- /J
.-
. ,.". . ,
a
+-M
y
FIG. XV. l.IFE-CYCLE OF Hydrodictyol/ reliClt/olulII (1..) LAGERHEIM

a-b, GAMETES; FIG. XVI. LIFE-CYCLE OF Chiorochylrillll/ [el1ll10e COliN

c-ci, ZYGOTE;
d, FOUR SWARMERS FROM ZYGOTE; a, ZYGOTE: b, PENETRATION OF ZYGOTE (s) INTO TilE HOST;
e, POLYHEDRON STAGE;
C, RESTING CELLS (r) IN LEAF OF £ell/I/O; d, suOWING STRUCTURE OF RESTING
f, NET FORMATION WITHIN POLYHEDRON;
CELLS; e-g, DIV1SION STAGES OF SAME; h, LIBERATION OF SWARMERS (g') Ir-:TO
g, SWARMER FORMATION WITHIN SAME;. VESICLE (b), m, OUTLINE OF MEMBRANE OF RESTING CELL; i, SWARMERS.
h, SMALL PORTION OF YOUNG COLONY (FROM TILDEN, 1935).

(d, AFTER BRISTOL; i, REDRAWN FROM KLEBS; THE REST AFTER

KLEBS).
 34 CHLOROCOCCALES ECOLOGY AND PHYSIOLOGY 35

intervals with special reference to the Chlorococcales (Philipose, 1959, and unpub!.).
Gonzalves and Joshi (1946) and V. P. Singh (1959) have also made some observations
on the ecology of some planktonic forms.
(A) Types of waters predom.inated. Generally most of the planktonic
Chlorococcales are observed in shallow ponds and tanks, 30 em. to It m deep.
Swamps with about 30 cm to a metre (or more) of water, paddy fields, and shallow
fisherybunqhs used for the semi-natural spawning of carps also appear to be favourite
haunts .of the Chlorococcales, a large number of them being occasionally noted in a
single collection. Other bodies of waters in which they occasionally predominate are
shallow rainwater ditches and pools, rock pools, small lakes, reservoirs and rivers in
which the water remains almost static from December to May. Concrete cisterns used
for experimental purposes also very often developed large number of Chlorococcales.
Moats and tanks with decaying vegetation also sometimes provide a peculiar type of
r ~ Chlorococcalean flora. W. & G. S. West (1902, 1907) have reported a number of
I~~j
I~~i <"1')..-
Chlorococcales from paddy fields, swamps, tanks, streams and artificial ponds in Ceylon
and Burma. Lemmermann (1907) and Holsinger (1955) recorded a few from the
I!B1~
, '. J
Colombo Lakes, and Cro\v (1923) observed a number of.th~m in rock pools, tanks and
lakes. Carter (1926) collected them in large numbers from ditches connected with
g F.
paddy fields and from streams in N.E. India. Handa (1927) and Skuja (1949) record-
M
ed quite a nu.mber of Chlorococcales from ponds and lakes of Burma.
Planktonic Chlorococcales are us~ally rare in brackish and salt water. However,
Moens (1958) has reported that a number of species like Ankistrodesmits Jalcatus,
Coelastrum microporum, Cr..ucigenia emarg~nata, DictyosPhaerium ehrenbergianum; Kirchrzeriella
. lunaris, Oocystisgiga$, O. submarina;Pediastrumboryanum,P. duplex, P. integrum, Scenedem,us
obliquus, S. quadrfcaudaand Tetrastrum multisetum could tolerate varying degrees of sali-
nity obtaining in the North Sea Canal, which he investigated. Apart from the occa-
sional presence of species of Ankistrodesmus, -Crucigenia, Dictyosphaerium, Oocystis,
Pediastrum,Scenedesmus,Selenastrum, Tetrastrum and Telraedronin tidal rivers (Iyengar and
~~ ~~~ Venkataraman, 1951; Dutta et al., 1954), no really.brackish water 'species have been re-
\q~J(1@Ji .. ported from the Indian r_egio.n. Among the very few marine Chlorococcales kn(\wn,
1- . ..~!.~~..:<;S:'? 9.~ Chlorellaalone has been recorded from the Indian region (Svedelius, 1907).
'P.7,./.:->-.,." :~{".: \:... '.
~~l._-'~~:::::~6 A~ _ ).~. (B) Species obse~ed in bloom.s. Though plankton collections from the--
. '~9'-''''7i )1 <.it;'t'~( I~<J~.? shallow waters in India referred to above invariably contained one or more of the
19:.~( \::.~:.j! >/Qf)",::~~"-~r11
,"
\'. . .,~.\::.~:-~~
. \'IJ ~...~ '\\' - ..~i>£:::_"1:~
./..... ~.,.
Chlorococcales, usually none of them occurred in very large numbers to constitute an
~~C~'T \". M' almost exclusive bloom. However, some species like -Cloenkinia radiata, Micractinium
\ifl'1:~ j~~~~) I
.~~..~::::J ~&~~/ .pusillum, Tetraedron minimum, Selenastrum gracile, Pediastrum simplex val'. duodenarium,
'i!-....
P. duPlex val's. clathratum, reticulatum and gracillimum, Botryococcus spp., Coelastrum
FIG. XVII. ADAPTATIONS FOR PLANKTONIC HABIT microporum,Scenedesmusbijugatus,S. quadricaudaval's. quadrispina,longispinaand westii were
occasionally found in such large numbers that they imparted a distinct green colour to
a, Tetraedronlimneticum BORGE.; b, ScenedesmUJ quadrieaudaVAR. longispina the water. Others, though not so abundant, still occurred in fairly large numbers in
(CH~D.) G. M. SMITH; c, RadioeoeeUJ nimbatus (DE WILD.) SCHMIDLE;. some waters, often mixed with other algae. Instances of these were Lagerheimia
d, Eehinosphaerella limnetiea G. M. SMITH; e, Seenedesmlls aeuminatUJI (LAGERH.)
CHODAT (SHOWING MUCILAGE BRISTLES); r, Franeeia ovalis (FRANCE) LEMM.;
g, Crueigenia tetrapedia (KIRCHN.) WEST; h, Marthea tetras PASCHER;i, Treubaria -+ (a-b, ORIGINAL;c, AFTERDE WILDEMAN;d-k, FROMG. M, SMITH, 1950;
blanetoniea (G. M. SMITH) KORSH.; j, Pediastrum simplex VAR. duodenarium e-j, AFTER PETERSEN (e, AS Pediastrum clathratllm (SCHROEDERLEMM.); r,
(BAILEY) RABENH. (SHOWING MUCILAGEBRISTLES); k, Micractinium pUJillum AFTER FRANCE; g, AFTERSCHMIDLE;i, AFTERG. M. SMITH, 1923 (AS Borgea
}IRES.; I, PeetodiclYon eubicum TAFT. . -+ plantoniea G. M. SMITH); I, FROMTAFT).
36
, though not very abundant, 'very often became dominant during this period.
, FIG. XVIII.
a, Fraru:tia drotsehtri (LEMM.) G. M. SMITH SHOWING MUCILAGINOUS SHEATHS
0' BRlSTUSj b, Tetrachlortlla
C, DtSIII/llraclflm dtlicalissimum
sphDer/l'slIdlllea (LEMM.) CHODAT; g-h, Siderocystris fusea KORSH., SHOWING AUTO-
SPORB 'ORMATION.
ALL AFTER KOR'lHIKOV, 1953).
CHLOROCOCCALES
ECOLOGY AND PHYSIOLOGY 37
chodati, DictyosPhaeriumpulchellum, Pediastrum araneosum var. rugulosum, P. duPlex vars.
coronatumand subgranulatum, P. muticum var. longicorne,Coelastrumreticulatum, C. scabrum,
Crucigeniafinestrata, Hofmania lauterbornei, Scenedesmusopoliensis and S. perforatus.
(C) Seasons of abundance. The majority of the Chlorococcales observed
by the author occurred during the sultry south-west monsoon period (June to
September), when the mean air temperatures usually ranged from 26'50 to 29'5°C
(Table I), the monthly sunshine hours were lowest (120-180) and there was a rain-
fall o~ ~bout 100-175 em. Some of these algae also showed a secondary maximum
during summer (March-May), when the mean air temperatures ranged from 26'5° to
32' 5°C, the sunshine hours from 220-310, and the rain fall from 7' 5 to 30 em, rarely
more. There were, however, a few species which occurred in abundance during the
cool months, particularly from November to February, when the mean air tempera-
'/0 ture usually ranged from 20° to 25' 5°C., the sunshi~e from 120-230 and the rainfall
from 12' 5 to 62' 5 em.
Sometimes; the altitude made a difference in the seasons of occurrence of some of
the species. Thus, in hill stations like Coonoor and Ootacamund (2,000-2,350 m),
:t where the average temperatures obtaining in s,ummer were less than what they were
in the plains during winter, a few species which occurred in abunaance in the plains
during winter attained their maxima only il\ summer.
The Chlorococcales which were usually predominant during the south-west mon-
soon months of Jl!ne to September were Tetraedron minimum, Lagerheimia. chodati,
P~hycladon .umbrinus, Pediastum simPlex var. duodenarium, Codastrum microporum,
DictyosPkaerium'pulckellum, Scenedesmus armatus var. bjcaudatus, & quadricauda var.
longisPinaand, rarely; S. opoliensis and S. tropicus. A number of others like .Vlicractinium
pusillum, Schroederia indica sp" nov., Actinastrum hantzschii, Pediastrum duPlex vars.
" reticulatumand gracillimum,Coelastrumscabrum,C. proboscideum,Crucigenia rectangularis,
C. aPiculata, C" tetrapedia, Westella botryoides, Scenedesmusdimorphus and- S.' denticulatus,
-A number of the algae mentioned above sometimes reappeared in fairly large
numbers 'in early or late' summer. Some like Pediastrum simplex var._ duodenarium,
DictyosPhaeriumpulchellum and Micractinium pusjilum be~ame even abundant occasionally.
A few others like Sorastrum spinulosum,Pediastrum tetras; Coelastrum,cambricum'var. inter-
m!dium, Crucigenia fenestrata, Hofmania laute~borneiand Scenedesmus quadricauda var.,
longisPina were 'also found dominant in sOIJ..lewater~ during summer. Of these,
'" Sorastrumwas rarely observed. during other seasons.
The species which appeared to become dominant during the cool (and usually
dry) months of October to February were Kirchneriellalunaris, NePhrocytiumagardhianum,
'N. lunatum, DimorPhococcuslunatus, Dictyosphaerium ehrenbergianum, Selenaslrum gracile,
ADAPTATIONS FOR PLANKTONIC HABIT
Pediastrumduplex vars. coronatum,reticulatum,clathratum, and gracillimum, P. araneosumvar.
rugulosum, P. muticum var. longicorne,Coelastrumreticulatum and Botryococcusprotuberalls.
ornata KORSH.; d, Micraetinium apptndiculatum ,KORSH.;
KORSH.; e-d, indutum (GEITLER) PASCHER; f, Botryo- oof Scen.edesmusaabundans. Some of these species like Kirchneriellalunaris and Dictyosphaerium
ehrenbergianumwere observed in large numbers in waters at high' altitudes during
NOTE THE MUCILAGINOUS ENVELOPES IN b,c, e, f. Summer instead of winter. .:;;'
(D) Physico-chemical features of the water and the Chlorococcales.
Apart from the differences in temperature, sunligl}t, rainfall and water level brou~nt
...
 CHLOROCOCCALES ECOLOGY AND PHYSIOLOGY 39
38

P. muticum val'. longicorne,Dictyosphaeriumehrenbergianumand Scenedesmus arcuatus val'.

about by seasonal differences, the physico-chemical features of the water appeared to
capitatus under temperatures ranging from 19° to 25°C.
have a great influence on the relative frequency and composition of the Chlorococca-
lean flora. The physico-chemical features studied were, the water temperature, In this connection it may be mentioned that Petersen (1946) has reported species
turbidity, pH value, total alkalinity and the NIP ratio, the nitrate nitrogen alone being of Scenedesmusfrom hot springs of Kamtchatka, where S. dmticulatus occurred at 50°C,
taken into consideration. S. quadricaudaat 29°C, and S. rostrato-spinosus'at 63°C. According to Elenkin .(1914,
The physico-chemical features of the waters in which the Chlorococcales cr., Petersen, l.c.), S. quadricaudahas been found even in waters with a temperature of
up to 53°C.
predominated were by no means uniform. Thus, 12 bodies of water from different
TURBIDITY. High turbidity or even muddiness of the water appears to be
parts of north-east and south India (Table-II), in which the Chlorococcales were do-
minant, showed great divergence in the ,relative frequencies and composition of the favourable for the development in large numbers of many Chlorococcales, the effect
flora and in the physico-chemical features. The maximum number (53 species) of of high turbidity b~ing that sunlight does not penetrate deep into the water thereby
Chlorococcales in a single collection was obtained from a swamp in Puri district and the simulating conditions of low sunshine existing during the sultry monsoon months of
next maximum (45 species) from nursery pond No. 38 at Cuttack. The shallow water the year. The turbidity in most cases depends on the rainfall and water level, shallo,~
of the former was neutral and with very low total alkalinity (32), NIP ratio was less waters often becoming turbid even when there is no rain, but sometimes other algae
than unity (0' 52) and the period of collection was April, when the water was very
also contribute to the turbidity. .
warm. In the latter, the water was more or' less alkaline (ph-8'4, T.A.-86), the NIP Tetraedron caudatum, Dictyosphaeriump~chellum, Pediastrum tetras val'. tetraodon,
ratio was less than unity (0' 06) and the period of collection was August, when the water Crucigeniafenestrata, Hofmania lauterbornei,Coelastrumreticulatum, and Scenedesmusopoliensis
temperature was moderately high. 'The remaining waters (alkaline, neutral or slightly
_ were all collected in large numbers from waters which were very muddy (Iyengar and
Ramanathan (1940) also collected DiclJosphaeriumindicum from muddy rainwater pools
acidic) supported ~arying numbers of Chlorococcales ranging from 13-21 species and
at Madras). A number of other founs like Schroederiaplanctonica, Tetraedron miniTr''''',
varieties, and the difference in the flora appearecl to be mainly due to the varying
T. limneticum, T. muticum,Lagerheimia chodati, Micractinium pusillum, Coelastrummicroporum,
physico-.chemical features, Thus, in a series of nursery ponds situated side by side at
Actinastrumhantz.schii,Ankistrodesmusspp., Westella botryoides,- Pediastrumsimplex val'.
Cuttack' (only five of these are referred to in Table II) those with moderately low al-
duodenarium,Crucigenia apiculata, C. tetrapedia,'Scenedesmusabundans,S. quadricauda vars.
kalinity (36-78) showed a flora somewhat different from those of ponds with higher
longispina' and -westii .also appeared to be favoured by moderately high turbidity
alkalinity (80-140). -No doubt there was a certain measure of'overl'!Pping in the t:ase
(50-250 ppm). However, a few Chlorococcales like Kirchneriellalunaris, Dimorphococcus
of some' species, indicating a certain degree of toleranc~ to varying physico-chemical
lunatus, NePhrocytium lunatum, N. agardhianum, P.' duplex and its varieties, P. boryanum,
features by these species. Dyke's tank, Visakhapatnam, with a high concentration
Coelastrumscabrum, and Scenedesmusartuatus val'. capitatus were usually observed in fairly
of nitrates (10 ppm), high NIP ratio (77'9) and fairly high a~kalinity (135 ppm) clear waters.
showed mostly ScenedesmUs spp. On the other hand, the difference in flora between
pH ANDTOTALALKA~INITY. Distinctly alkaline waters (total alkalinity well over
the swamp in Puri and the bundh in Chandrakona Road; Midnapore, in which the
100 and pH above 7' 7) were found suitable for the occurrence of certain Chlorococcales
physico-chemical features were somewhat similar, was apparently due to the seasonal
in large nUJ!1bers. Schroederiaindi;a, NePhrocytiumagardhianum,Actinastrumhantz.s,hii,
.
I
_ factor alone, the collection from the swamp being taken in summer and the collection,

from the bundh in winter. '

"
-

By comparing the physico-chemical features--of'waters from different parts of

India in which one and the same species of Chlorococcales predominated (Table III),
it has been possible to arrive at certain tentative correlations between some species
and certain physico-chemical features of the water.
WATER TEMPERATURE.A number of species and varieties like Micractinium
pusillum, Tetraedron minimum, Treubaria triappendiculata,Lagerheimiachodati, Ankistrodesmus
falcatus, Kirchneriellacontorta,Pediastrum simPlexval'. duodenarium, P. duPlex var. subgranula-
tum, P. biradiatum val'. longecomutum,Scenedesmusacuminatus,S. opoliensis val'. mononensis,
S. carinatus, S. quadricaudaval's. longispina and westii have been observed in waters with
the temperature ranging from 30° to 37°C; Ankistrodesmusspiralis, Kirchneriella lunaris,
Dimorphococcus lunatus, Pediastrum duplex vars. clathratum, reticulatum and gracillimum,
Crucigenia tetrapedia, Scenedesmustropicus, S. acutiformis and Tetrallantos lagerheimii were
observed-in fairly large numbers in waters with the temperature ranging from
250 to 30°Cj and NePhroCJtiumagardhianum, N. lunatum, Pediastrum duPlex val'. coronatum,
PediastrumsimPlex val'. duodenarium,fi: muticum val'. longicorne,Coelastrummicroporum,Soras-
trum spinulosum,Scenedesmusdimorphus,S. acutiformis, S. abundans, and S. quadricau4aval'.
longispina are some examples. Moderately alkaline waters (total alkalinity 60-100,
pH 7' 7-8' 5) appeared suitable for the development of a number of other forIns like
Micractiniumpusillum, Tetraedron minimum, T. limneticum,Lagerheimia chodati, Botryococcus
braunii, Nephrochlamys subsolitaria, Pediastrum tetras val'. tetraodon, Coelastrum proboscideum,
Crucigeniaapiculata,Scenedesmusdenticulatus, S. opoliensis, and Tetrastrum punctatum. On
the other hand, neutral or slightly acidic waters (total alkalinity 8-48, pH 6'2-7'1)
appeared more favourable for the development of species like Treubaria triappendiculata,
Nephrocytiumlunatum,Ankistrodesmusfalcatus, A. spiralis, Kirchneriellacontorta,DictyosPhaerium
ehrenbergianum, Dimorphococcus lunatus, Pediastrum biradiatum.,' Scenedesmus acuminatu.r,
Tetrastrum heteracanthum,Tetrallantos lagerheimii, and Botryococcusprotuberans.
Mention may be made here of a few species of the Chlorococcales from outside
India reported mostly from acidic waters or even acid bogs. The free-living Eremo-
sphaera viridis and Desmatractum bipJramidatum (Lund, 1942), the epiphyte Octogoniella
40 CHLOROCOCCALES
and the endophyte Phyllobium sphagnicola are typical examples. According to Lund
(l.c.), the substances associated with low pH in nature may also be an important factor
determining the conditions of occurrence of Desmatractum. DimorPhococcuslunatus
(Prescott, 1951) is also known to occcur in somewhat acidic waters.
NIP RATIO. A number of Chlorococcales (Table III) occurred abundantly in
waters with an NIP ratio less than unity whereas a number of others were observed
when the ratio was higher than unity. Thus, Schroederiaindica, Microctiniumspp.,
Tetraedronminimum, T. limneticum,Lagerheimia chodati,Actinastrumhantzschii, Pediastrumsim-
plex var. duodenarium,P. duplex var. subgranulatum,P. araneosumvar. rugulosum,Nephrocytium
lunatum, Coelastrum microporum, Scenedesmus dimorPhus,S. denticulatus, S. opoliensis and
Tetrastrum punctatum occurred in waters with an NIP ratio usually less than 'unity
whereas, species like NePhrocytiumagardhianum, Kirchneriella lunaris, Scenedesmus asmatus
Dimorphococcus lunatus, Pediastrum duplex vars. coronatum, clathratum, reticulatum, and
gracillimum, P. muticum var. longicorne,Botryococcusprotuberans,Coelastrum cambricum var.
intermedium, Crucigeniatetrapedia, Scenedesmusabundans, S. perforatus, S. quadricauda vars.
langispina and westii, and S. acutiformis occurred in waters with the NIP ratio usually
more than unity. A number of others like Selenastrum gracile, Ankistrodesmus spiralis
Dictyosphaeriumpulchellum, Coelastrumscabrum, and Tetsallantos lagerheimii appearc:d to be
more or less indifferent whether the ratio was higher or less than unity.
Golenkinia radiata, Scenedesmusbijugatus yar. flexuosus, S. quadricaudavars. longispina
and quadrispi1}a,and Coelastrumscabrumalso came up in profusion in cement cisterns which
had fairly high concentrations of ammoniacal nitrogen (and presumably with, NIP
ratio higher than unity) followingthe killing of submerged macroflora by the applica-
tion of urea at 50-250 ppm. ~ also. ,The intense sunlight obtaining in shallow waters during bright periods is very
According to Ryther (1954), who studied the ecology of phytoplankton blooms
in a bay enriched by duck farm wastes, by simultaneous field and culture studies,
green algae of fresh and brackish water grow much mor~ rapidly in water containing
three times as much phosphorus per atom of nitrogen as normal sea water, the normal
ratio of total' nitrogen to phosphorus in sea'water being 15: 1._ In the present observa-
tions, in which the Chlorococcales alone' were considered, some algae aRpeared to be
favoured by an NIP ralio (nitrate nitrogen alone taken into considerationf higher Jhan
unity, wh~!eas others appeared to be. favoured by a ratio less than unity. Natarajan'
(1959) reported an NIP value of 22Ll as necessary to obtain maximum grow~h of
Selenastrum westii in cultures, as against 15/1 reported for NePhrochlamys subsolitaria
(~Kirchneriella subsolitaria) by Potash, 1958 (Natarajan, l.c.).
RESIDUAL FACTORS INFLUENCING THE PREDOMINANCE OF THE CHLOROCOCCALES.
A number of other factors not mentioned above are also probably responsible for the
occurrence and abundance of the Chlorococcales.
According to Kolkwitz and Marsson (cf. Brunnthaler, 1915), Chlorococcuminfusio-
num is characteristic of highly organic waters; C. botryoides(now considered as a member
of the Xanthophyceae under t4e name Chlorobotrysregularis), C. hum~colo,Dictyosphaerium
ehrenbergianum,D. pulchellum, Pediastrum ~oryanum,Ankistrodesmusfalcatus val'. acicularis,
Scenedesmusacuminatus,S. obliquus, and S. quadricaudaare characteristic of waters with
medium organic content; and Dimorphococcuslunatus, Ankistrodesmusfalcatus, Westella
botryoides,Pediastrum duPlex, P. kawraiskyi, P. tetras, P. biradiatum, Actinastrum hant;:,schii,
ECOLOGY AND PHYSIOLOGY 41
CoelastrummicropiJrum,C. reticulatum, and Hydrodictyon reticulatum are characteristic of
waters which are fairly pure, but with dissolved mineral substances and a little organic
matter.
Chick (1903) reported that Chlorellapyrenoidosaoccurred in sewage polluted water.
Chlorellaand Scenedesmusare also commonly knO\yn to occur in sewage stabilization ponds
(Raman, 1959).
Though no determinations of the organic matter of most waters studied by the
author were ma<;le,it was observed that some of the Chlorococcales dominated in waters
in which the abundant aquatic weeds present were killed by chemical treatment or in
which the weeds were naturally decaying. Thus, forms like Golenkinia radiata,
Micractinium pusillum, Ankistrodesmus sPiralis, Selenastrum gracile, Pediastrum tetras val'.
tetraodon, Sorastrum spinulosum, Coelastrum cambricum val'. intermedium; C. proboscideum,
C. scabrum,Crucigeniatetrapedia, Scenedesmusbijugatus var..flexuosus, and S. quadricaudaval's.
quadrisPinaand longisPinawere frequently found occurring in fairly large ~umbers in
such waters.
It, thus,' appears that the_composition and, relative frequencies of the Chloro-
coccales occurring in inland fresh-waters c!-ependon a number'of factors which ,operate
severally or collectively. Of these, the rainfall, temRerature, sunshine, water level, and
turbidity; pH, total alkalinity, the nitrogen-phosphorus ratio and, probably, the
dissolved organic matter appear to be important.' It is also fairly clear that a large
number of Chlorococcales show their maxima during the sultry south-west monsoon
E.eriod', l>articularly during the months of July and August, very often with secondary
.
maxima iri summer. A few forms. become preaominant during the winter months
often off-set by the turbidity of the water. 'Neutr~l or slightly acidic waters support
a parjicular Chlorococcalean flora which are usually differen~ from those occurring
in alkaline waters. However, some species are tolerant of acidic as well as alkaline
conditions. In waters richer in nitr!ltes than phosphates some species of Chlorococcales
appear to thrive better and vice versa.
Fritsch (1907), who studied the flora of the tanks, lakes and other inland waters
of Ceylon" observed that the algal ,flora of tropical waters are greatly inft'fIenced by
the illumination, temperature, frequent changes in water level and to some ex:terit by
the chemical composition of the disso1ved matter, degree of movement of water, nature
of the substratum' and the muddiness of the water. According to him, apart' from
""',f
the frequent changes in water level brought about by rains and ,heat and which affect
the concentration of dissolved salts, the daily temperature range of 6°_10°C and the
intense illumination taken along with turbidity (caused by suspended mud particles
or by algal blooms) are the most important factors which determine the flora of stag-
nant waters of the tropics. Though his study was mostly with reference to the blue
green algae and green algae other than the Chlorococcales, his observation.s appear to
hold good in the case of the Chlorococcales as well, which form an important consti-
tuent of tropical fresh-water algal flora. In addition, it appears that the seasonal aspect
and some of the physico-chemical features of the water, which Fritsch could not study
owing to the short duration of his observations, are also equally impOl;tfmt in deter-
mining the type of flora. .
 ECOLOGY ANI) PIIYSIOLOG'" 43
42 CHLOROCOCCALES

Attached forms. There are three main categories of attached Chlorococcales:

(a) attached to macroflora and filamentous algae, or epiphytes; (b) attached to
rotifers, copepods, cladocera, mosquito larvae or other animalcules, or epizoic (or
epizootic) forms; and (c) attached to stones, pebbles and rocks, or lithophytes
(Fig. XIX).
Epiphytes: Most. species of Characium and allied genera like Pseudocharacium,
Pseudochlorothecium,Hydrianum and Hyalocharaciumocc~r epiphytically (attached by stalks)
the substratum being usually filamentous algae,' rarely macroflora. Desmatractum
bipyramidatumand Octogoniellasphagnicolaare two good instances of algae belonging to
the C,hlorococcales which occur attached to the leaves of aquatic macroflora, the
c
former by its outer mucilaginous envelope and the latter by its flat side. Ectogeron
elodeaealso spreads itself flat on the leaves of submerged aquatic plants. Pulvinococcus
and BicusPidella, on the other hand, are fixed with the aid of a gelatinous cushioq or
short thick stalk.
" In the Indian region, Characium nasutum has been found on Oedogonium (R.N.
~ .'
Singh, 1939) and on Cladophora(Skuja, 1949); C. ambiguumon PithoPhora or Oedogonium
(Dixit, 1937; G. S. Venkataraman, 1957); C. angustum on decaying algae (Turner,
1892) and C. apiculatum on Chaei()tnorphaand Schizomens (Skuja, 1949). Trochiscia
granulata val'. aeroPhila (Skuja, l.c.) has been reported as occurring on Euglena.
Sometimes certain .pedes of the Chlorococcales are found attached to floating
stems and twigs. Thus ChlorocoCGum vitiosum has been found (Skuja, 1949) ~on
floating stems of bamboo in a pond and Characiosiphon'has been found (Agarkar; 1953)
occasitmally on floating twigs, 'leaves or even pebbles. .

Ep{zoic forms (Fig. XIX d~, I): A number of Chlorococcalean genera like
KorshikoviellaSilva, RhopalosolenFott, GloxidiumKorshikov, HyaloraphidiumKorshikov, and
t.- C~araciumA. Brau~ are ~nown to occur on various,animalcules like Cyclops, Branchypr:s'
s,r;jj
Ii '" Dlaphanosoma,Moma, rotlfers and even on AnoPheleslarvae. Amongst these, the specIes
Korshikoviellagracilipes has been observed by the author on Moina in a nu~ber of ponds
in Orissa. K, limnetica which ha.s been reported as occurring on DiaPhanosoma.01' !n
the' free floating condition (Skuja, 1948) has 'also been observed, but only in the
. detached condition. CharaciuTIJ
debaryanum
has been observed by the author on rmifers,
rarely on Cyclops. Another Indian species is C. anophelesiwhich occu'rs on sev(:ral
species of' AnoPheles ~arvae. Apart from the fact that all these species are attached,
there is probably some sort of symbiotic association between the animalcules and the
algal species concerned, as suggested by the usuJI occurrence of a' particular species' of
alga on a particular type of animalcule. It is also clear that one of the conditions. for
g h k
the predominance of these algae is the presence in large numbers of the animalcules
concerned in the plankton.
FIG. XIX. LITHOPHVTIC, EPIPHVTIC, EPIZOIC, AND ENDOPHYTIC FORMS
LithoPhytes: Characiosiphon and Dendrocystis are both examples of lithophytes.
a, CharaciosiPhon rivularis IVENG.,CLUSTER OF PLANTS ON A PEBBLE; b, Characium
SP. ON Oedogonium; c, C. prinsheimii A. BRAUN ON Tabellaria flocculosaj d, C. anophelesi .T~e former occurs in dusters on stones and pebbles (Fig. XIX a) in shallow streams
!VENG. ET !VENG. ON THE APPENDAGE OF A MOSQUITO LARVA; e, C. debaryanum (REINSCH) (Iyengar, 1936), but; rarely, on other submerged strata as well. The latter remains
DE TONI ON THE ROTIFER Brachionus; f, Octogoniella sphagnicola PASCHER ON FRAGMENT "
OF A SPHAGNUM LEAF; g-k, Chlorochytrium dinobryonis LUND INSIDE Dinobryon divergens;
attached to stones of streams in the initial stages as well as in the dendroid condition.
g, CELL DEVELOPING INSIDE THE LORICA; h, FULL-GROWN CELL (st, STARCH MASSES);
i. FORMATION OF ZOOSPORES: j, EXTRUSION OF PROTOPLAST FOR LmERATION OF ZOOSPORES:
~ (a, AFTER IVENGAR, 1936; b, c, FROM G. S. WEST, 1916; f, AFTER PASCHER;
k, WITH APLANOSPORES; I, Korshikoviella gracilipes (LAMBERT) SILVA ON THE CLADOCERAN g-k, FROM LUND, 1955; m, FROM CUNNINGHAM, 1888 (AS Stomatochytrium limnanthemum
Moina; m, Chlorochytrium limnanthemum (CUNNINGH.) G. S. WEST INSIDE Limnanthemum CUNNING.); e, FROMPHILlPOSE, 1940; 1,X 725). .
indicum LEAF TISSUE AND SHOWING TWO ZOOSPORANOIA. ~

44 CHLOROCOCCALES
Rocky surfaces with dripping water often show a well defined association of algae,
sometimes with Trochiscia and Oocystis (Tiffany, 1951). In India, Oocystis elliptica,
I
O. solitaria (Biswas, 1934) and O. kumaonensis(K. P. Singh, 1960) have been recorded
I from dripping rocks, often embedded in the mucilaginous masses of colonial blue
green algae.
I Cryobionts. It has already been mentioned elsewhere that some of the
Chlorococcales form snow floras. Of these, Scotiella and some species of Trochiscia
are the most prominent. Chionaster,Mycacanthococcusand, rarely, Ankistrodesmus and
Tetraedronalso occur on snow and ice. Species of the foregoing genera are known from
the Antarctic and Arctic regions or from the snow-clad mountains of Europe, North
and SQuth America. They are usually associated with unicellular Volvocales,
Zygnemales and the Myxophyceae. Fritsch (1912) and Kol (1942) have given
good accounts of the snow and ice floras of the south Orkneys and Alaska
respectively.
According to Kol (l.c.), the cryobionts live very close to the surface and, so, the
changes taking place on the surface of the snow exert the greatest influence on these
organisms. Since the source of mineral substances for these organisms is the surrQund-
ing rocks, the nature of the rocks (whether composed of lime-stone or acidic) would
decide the nature of the flora. Kol divided cryobionts into four classes, viz. 'glacialis- '
cryobionts ' or those which occur only on ice, , nivalis-cryobionts ' or those which occur
only on snoW, , mixo-cryobionts ' or those which are adapted to both snow and ice and
, cryoxen ' or those which have their normallocabon on damp cliffs but get transferred
to ice and snow. Trochiscia cryoPhilaf. longispina-Kol and brevispinaKol are examples
of -. glacialis-cryobionts '; Scotiella nivalis, S. antarctica,-Mycacanthococcus cellaris-f.
antarctica, M. ovalis, and Tetraedron valdezii are examples of · nivalis-cryobionts-';
Trochiscia antarctica and T. nivalis .are examples of · mixo-cryobionts' and Scotiella
po/ylJterais an example of a ' cryoxen '. Autospore formation appears to be the best
mode of reproduction in these cryobionts.
Species of Scotiella give an orange red to yellow colour to the snow. S. polyptera
and Mycacanthococcuscellaris f. antarctica Wille ~ave also been observed in green snow
(Kol, l.c.). _ - .. - ., -,' -
No cryobionts have so far been recorded from the Indian region. However, the
long range of the Himalayas with its perpe~ual snow and .ice is bound to show a
number of these cryobiont~ including those pelonging to the Chlorococcales.. - the barks of Anacardium occidentaleand Albizzia labbek in Burma.
Terrestrial or aerophilous .forms. There are also members of the Chloro-
coccales which occur in diverse terrestrial habitats. Thus, several members of the
Chlorococcaceae, including Chlorococcum humicolo,and C. hypnosporum - and of the
Chlorochytriaceae like KentrosPhaerabristolae, are usually found in soils,' cultivated or
otherwise, or, rarely, species of Chlorococcummay be found in stagnant waters also.
These algae seem to be capable of tiding over unfavourable conditions of weather
like drought in the soil, since they come up in cultures of dried soils. Tamiya (1959)
reported that a Chlorella survived in a sample of desiccated soil for over ten years.
Adaptation to desiccation is also seen in a number of Chlorococcales like Myrmecia
globosa, Phaseolaria obliqua and .Chlorococcumvitiosum which are all aerophilous forms
occurring on the barks of trees.
ECOLOGY AND PHYSIOLOGY .l5
Palmellococcusminiatus (=Chlorella miniata) and Chlorella conglomeratahave been
reported (G. S. West, 1916; Skuja, 1949) as' pale green patches on flower pots.
Hansgirg's Mycacanthococcuscellaris, Mycotetraedroncellareand Myurococcusurococcusare all
known from damp walls. Palmellococcus protothecoides (=Chlorella protothecoides),
Palmellococcussaccharophilus(=Chlorella saccharoPhila),Protothecamoriformis and P. zopfii
usually occur in the exudations of sap from trees (Brunnthaler,1915; Printz, 1927),
though Palmellococcussaccharophilus is occasionally observed on flower pots (Sukja,
1949). Petersen (1928) has reported several Chlorococcales like Myrmecia pyriformis,
Trebouxia arboricolaand Chlorella rugosa from wooden beams and other wood works in
Iceland. Trochiscia granulata var. aeroPhila is another aerophilous alga known from
sub-aerial situations in Durban along with Phaseolaria, Chlorococcumvitiosum, etc.
(Printz, 1921), but found by Skuja (1949) in Burma growing on Euglena. Rhodochy-
trium occurs epiphytically (or endophytically) on the land plant Ambrosia.
Apart from Chlorella conglomerata, Palmellococcussaccharophilus and Trochiscia
granulata var. aerophila.reported from terrestrial habitats in Burma by Skuja (l.c.), a
number of Chlorococcales have been reported from cultures of soils from the Indian
,. ;.. region. These include Chlorococcum humicolo(Biswas, 1934; Gonzalves and Gangla, 1949),
C. infusionum (R. N. Singp, 1939), KentrosphfWa bristolae (as Chlorochytriumparadoxum,
,/tt >:-R. N. Singh, 1939), Characium acuminatum (Singh, 1939), C. nasutum (Khan, 1957),
C. terrestris (Kanthamma, 1940) and Trochiscia aspera (Gonzalves and Gangla, 1949).
~.-
Algae like Chlorococcumhumicoloare also found in the rhizospheres of crop. plants
(Gonzalves et Yalavigi, 1959).
An interesting instance of an alga', which, though norma11y found in the plankton,
~ of ponds and rivers, can thrive healthily on moist sand near water, is Scenedesmus
arcuatusvar. capitatus. This alga was found by the author (Philipose, 1959) in the form
of extensive patches on the sandy margins of River Daya in Orissa during November
1952. The same alga was found in a sample (leg. Dr M. P. Motwani) collected during
the same month under identical conditions from River Dehri-on-Sone, Bihar.
Schroederia planctonica(=Characiumplanctonicum),which is a planktonic species, is also
found sometimes on ~et ground (Skuja, 1949). .
Several Chlorococcaleslike Trebouxiaand Chiorellahave also acquired a terrestrial
.- habitat in association with fungi to form lichens. These will be dealt with under the
symbiotic forms. Tribouxia has also been reported (as Cystococcus-Skuja, 1949) from
Endophytic, endozoic and parasitic forlDs. (Fig. XIX g-m, XXI a-d).
Endophytic forms usually occur within the tissues of higher aquatic plants, rarely with
in other algae. According to Fritsch (1935), it is only a short step from epiphytism
to endophytism. Fritsch also stated that it is not clearly established whether genera
like Ch~orochytriumare mere space parasites which damage the tissues of host plants in
their immediate neighbourhood only or whether they derive any nourishment from
them. Since individual species are very often found confined to particular hosts, he
considers some kind of interrelation between the two quite probable. In forms like
Rhodochytriumthere is parasitism accompanied by loss of chlorophyll and, though the
,damage to the host is not appreciable, the phloem is frequently locally damaged,
. Iyengar
. (1951) referred to Chlorochytrium
as a space parasite and to Phyllobium
'..'
46 CHLOROCOCCALES
dimorphum, P. sphagnicola and Rhodochytrium spilanthidis as true parasites (also see
Oltmanns, 1923).
The endophytic genera are all zoosporic and most of them belong to the family
Chlorochytriaceae. Chlorochytriumlemnae is a common species in which the zoospores
or motile zygotes produced by sexual fusion settle down on the leaves of Lemna. A
cell wall is developed and the cell produces a tubular .prolongation into the host
through a stomium or in between two epidermal cells. Once inside, the tube swells
up in the intercellular space, where it forms a large ellipsoidal to lobed cell which
receives the protoplasmic contents from the cell settled outside. Then it forms a resting
cell with thick and stratified wall having local excrescences. When the Lemna dies
during an unfavourable season and falls to the bottom mud, the resting cell of the
alga is also carried with it. When the conditions are favourable again, the dormant
resting cell gives rise to a large number of swarmers which escape by the rupture of the
thick wall and the surrounding tissues of the host, the swarmers being enclosed within
a wide mucilaginous .envelope. The swarmers very often fuse within the envelope
or outside. The swarmers or the quadriflagellate motile zygotes infest a fresh host.
Chlorochytriumiemnaeoccurs as an endophyte, also on other plants like CeratoPhyllum
demersum, Elodea canadensisand some mosses. C. limnanthemum(Cunning.) G. S. 'West
(=Stomatochytrium iimnanthemumCunningham, 1887) occurs insid~ Limnanthemum indicum.
C. cohnii and C. sarcophyci(Fritsch, 1935) occur endophyti::.lly in the marine algae
Enteromorpha and Polysiphonia and within the mucilaginous envelopes of the diatom
Schiz;onema. Chlorochytriumdinobryonisis"another interesting species (Lund, 19.55) which
lives inside the empty lorica of Dinobryon divergens. KentrosphaeraJacciolae Borzi (incl.,
K. gloeophiia (Bohlin) Brunnthaler) sometimes occurs within the -mucilaginous en-
velopes of blue green algae. ApodochlorissimPlicissima(Korsh.) Komarek is found with-
in the mucilaginous envelopes of Microcystisand Aphaniz;omenon.
Phyllobiumdimorphumforms its resting cells along the main veins of Ajuga and
Lysimachia and P. sPhagnicolaon Sphagnum. RhodochytriumsPilanthidisoccurs on the leaves
of Ambrosia artimisiaefolia.
Some .species of CodWlum A. Braun, another endophytic genus, have been
considered as the sporophytes ~f Cladophorales while others are held under suspicion.
The thallus and reproductory structures of most of these algae are more or less
adapted for an endophytic or even parasitic habit. The Jhallus is'usually in the form of
a branching filamentous thread, which is coenocytic and it ramifies the intercellular
spaces or sometimes breaks down a few cells of the tissues of the host plants. Here
and there, resting cells or gametangia, with thick stratified walls are formed and
these give rise to zoospores or gametes.
A few Chlorococcales are aiso known to occur within the tissues of various
animals, but these are all probably symbionts and will be dealt with under that head.
Saprophytic fOnDS and forDls grown in cultures. A number of Chloro-
....
coccales, which are capable of a holophytic existence, exhibit a saprophytic tendency
when grown in artificial cultures or in media rich in organic matter, and in these
media they might even show better development (Fritsch, 1935). Thus, Chiorella
thrives in organic media like sewage, often accompanied by loss of chlorophyll.
Chiorella, Ankistrodesmus, Scenedesmus, and several others also grow very well on agar
ECOLOGY .\;>;U PHYSIOLOGY ,p
supplied with mineral solution and the growth is luxuriant if glucose is also present.
Fermenting cellulose is also useful in the place of glucose.
Some of the Chlorococcales can also be grown successfully in cultures in the dark.
Usually the algal cells become colourless or yellow when grown in darkness, though
they are quite healthy in other respects, the decolorization being attributed to the
inhibiting effect of glucose. When such algal cells are transferred to a medium having
a better balance of glucose and nitrogen, the cells might regain the green colour for a
short time. Some like ScClltdesmus obiiquushave also been grown in darkness for several
years without loss of chlorophyll. These algae are probably capable of producing
chlorophyll in darkness. Forms which live heterotrophically during darkness mayor
may not change to autotrophic nutrition when exposed to sunlight.
Quite a number of members of the Chlorococcales easily lend themselves for
culturing in the laboratory in mineral solutions containing the essential nutrient salts,
including micro-nutrients under suitable pH, temperature, and light conditions.
Chlorella,Selenastrum,and Scenedesmusare typical examples. Hopkins and Wann
(1926, p. 353) demonstrated that in cultures the full effect of changes in pH on
Chiorellais seen only when iron, which gets. precipitated at.pH above 5'7, i.<tkept in
solution by the omission of calcium and the addition of sodium citr~te. Under such
conditions the alga can grow at pH ranging fr!>m3'4to 7'~ with best growth at 'pH 7'5.
Natarajan (1959) found both Fe-EDT A and Fe-tartrate proved to be satisfactory sources
~
of iron for grow~ng Seunastrum westii in inorganic media. Hopkins (1930) 'has also
demonstrated the. importance of manganese in getting better growth of Chiorella .in
cultures. I --.
According: to Witsch (1959), apart from the necessary -nutrition ana- proper
illumination, a regular supply of carbon dioxide and proper circulation of water are
desirable for mass cultures of algae. Ten times. the normal supply of nitrogen in a
nutrient solution is also stated to be favourable for maximum production of green algae
like Chlorellaand Scenedesmus.
Chodat (1999, 1913) carried out pure culture studies of many Chlorococcales
and concluded that many genera like Ankistrodesmus,Dactyiococcus,and Scenedesmusshow
polymorphism (Fig. XX). - Chlorella-likestages have also been shown to occur iQ.certain
cultures of Scene.desmusand Goeiastrum._ According to Ohodat, !here are a number of
elementary species in a large number of genera of the Chlorococcales as shown by
differences between individuals grown in cultures, and these differences are morpho-
. logical: New elementary species, according to him, are formed by small mutatiom.
According to Fritsch (1935), it is very doubtful whether polymorphism exists in nature.
Further, studies in pure cultures alone do not give an idea of form variations in algal
species, since a given alga might very often assume a form in these cultures which
..
might be indistinguishable from another species. A distorted picture of the life-history
may also be obtained owing to abnormalities occurring in cultures. So, Fritsch-
considers pure cultures reliable and useful only in supplementing field observations.
Pure culture work has also been useful in showing that the gonidia of Tdbouxia
extracted from different lichens belong to distinct races. These differences in race
might be due to the influence of the fungal partner, considerable physiological changes
being brought about by the living together ,of an alga and a fungus. The alga ha~
 ECOLOGY AND PHYSIOLOGY 49
48 CHLOROCOCCALES

Giaucocystisis an interesting colourless member of the Oocystaceae (Fritsch, 1935;

.. :. .. Tiffany, 1951) inhabited by a symbiotic rod-shaped member of the Chroococcaceae

i9jJ
0~
::~ .~
~~.:: (Myxophyceae). Originally the curved radiating bodies inside the cells were taken
for the chloroplasts (Brunn thaler, 1915), but now they are definitely established as
the blue green component of an association between two algae (Fritsch, op.c.; G. M.
Smith, 1950). Since the blue green component does not appear to lead independent
existence, Fritsch included Giaucocystis under the Oocystaceae, whereas Smith, and
Prescott (1951) restricted, the generic name to the blue green component and included
the alga'tinder the Myxophyceae. Skuja (1949) placed it under a separate class, the
Glaucophyta, between the Cyanophyta and the Chlorophyta. The formation of
autospores within the cells and the possible formation of swarmers in Giaucocystis
(Fritsch, op.c.) have been advanced as reasons for not ignoring the importance of the
Chlorophycean component. .

,- a
c
b

".

FIG. XX. Ankislrodesmus falcalus (CORDA) RALFS GROWN ON AGAR-

..-. ...'8"
GLUCOSE, SHOWING POLYMORPHISM (FROM CHODAT, 1913).
a
been considered by some authors to lead a saprophytic existence, being supplied by
organic nutrient by the fungus, while the modern view is that it is an instance of
symbiosis.
The saprophytic. tendency has 'probably ied to the. origin of permanently
colourless forms like-- Hyaiocharacium, Prototheca, Hyaioraphidium, and MyciJtetraedron
(Fritsch~ 1935). . . ..
The endophytism of space parasites like Chiorochytrium and the occurrence of
green cells within animals have also been sometimes considered as an expression of the b g
saprophytic .tendencies of the Chlorococcales. . FIG. XXI. ENDOZOIC AND SYMBIOTIC FORMS
i.
S}'IDbiODtS. There are also Chlorococales which occur in symbiotic rela- a, LONGITUDINAL SECTION OF THE HYPOSTOME OF THE HYDROID Myrionema
tionship (Fig. XXI) with other plants or animals (Oltmanns, 1923). TrJbouxia and amboinensis, WITH CELLS OF Chlorella IN THE ENDODERM CELLS (en); b, Chlorella
FROM THE SAME; c-d, Chlorella IN Hydra viridis; c, CELL OF ENDODERM WITH
Chlorella are examples of genera which live symbiotically with various fungi to form a .-. ~ LIVING AND DISINTEGRATING GREEN CELLS; d, SECTION OF ENDODERM WITH PART
definite lichen thallus. Trebouxia occurring in the Parmeliaceae, Usneaceae and OF AN EGG BELOW INTO WHICH THE GREEN CELLS ARE SPREADING; e-f, GJauco-
cysiis nostochinearum ITZlGs.; . e, SINGLE CELL; f, FOUR-CELLED COLONY;
~oniaceae, and Chiarella (?) recorded in species of Ciadonia, all form lichen gonidia g, Cladoniafurcala BORNET, CROSS SECTION OF THALLUS.
(Fntsch, 1935). There is also probably a symbiotic relationship between Chiorella a, e, GREEN ALGAL .CELLS; b, DISINTEGRATING ALGAL CELLS; e,
ECTODERM; en, ENDODERM; N, NUCLEUS); e-f, (b, BLUE-GREENSYMBIOl<TS); G (u,
and the.ni~gen fixing:bacterium Azotobacter chroococcum(Lipman and Teakle, 1925). UPPER SURFACE 1, LOWER SURFACE).
According to Lazo (1961), Chiorellaenters into full association with Myxomycete (E-b, AFTER SVEDEUUS, 1907; c-d, AFTER BEIJERINCK (BOTH AS
plasmodia of several species which have been freed from bacterial contamination and "ZOOCHLORELLA") ; e,f, AFMER GEITLER and HlERONYMOUS RESPECTIVELY;
g, AFTER OLTMANNS, 1923).
grown on agar.
;')0 CHLOROCOCC,\L£S
Several members of the Chlorococcales live in association with animals and they
appear as green cells within the tissues of these animals (Oltmanns, 1923; Fritsch, 1935).
Freshwater animals like the Infusoria Stentor, Paramoecium, Ophyridium, some Foramini-
fera, the Coelenterate Hydra, the sponges Anodontaand Unio, mussels and snails, and some
Turbellarians have all been shown to contain such green cells, and in most instances
the alga concerned appears to be species of Chlorella,though other green algae might
also be found occasionally in some other animals. Scenedesmusquadricauda is a rare
instance of a colonial alga living symbiotically with the animal Carterius stepanowi.
Chlorella occurring within animals has been termed by Brandt (1882) as :(,oochlorella
with two common species, viz. :(,. conductrix and :(,. parasitica. Beijerinck (1890)
treated them as the respective species of Chlorella. Some authors consider these as
identical with C. vulgaris. However, Fritsch (op.c.) suspected different elementary species
in different hosts. - often found in the guts of various species of fresh and brackish water fish and they
The relationship between the animals and the plants appears to be a symbiotic
one in most instances. The food manufactured by.the alga is utilized by the animal
and the alga in turn gets a plentiful supply of carbon dioxide. Sometimes, a certain
num.ber of algaL cells are also digested by the animal. This symbiotic relationship
is not obligatory, since all the animals may not contain the algal cells and colourless
animals can thrive independently.
HmNever, in the case of the green algae occurring in the mantle and gills of
freshwater mussels and which are exposed to sunlight, the relationship between the
-alga and the mussel might very often 'range from true-symbiosis to parasitism (Fritsch,
1935). Link (1911) has described a parasitic green alga occurri~g within the skin of
carp. According to Fritsch (1935), it could probably be a species of Chlorochytrium.
How exactly the alga finds entry into the animal is not clearly established. A margins of shallow waters and occasionally in the plankton are sometimes encountered
number of algae are taken as food by these animals, but the special symbiotic alga in
all probability escapes digestion and establishes itself in _the tissues of the animal..
Sometimes, the reproductory cells like the gemmulae of Euspongilla and the ova of
H;'dra are infected, and the former is usually green. Motile juvenile stages of many
animals are also probably infected easily. Someti!iles, as in Stentor, the Chlorella cells
liberated from the dead anim_al-during' autumn remains in the plankton and reinfect
new animals during the following season..
Trochiscia zachariasii constantly occurs, in the mucus coveriI!-g the embryos and
larvae of the frog Rana, agilis. . OoPhila.amblystoml!tisoccurs witpin the:: envelopes sur-
rounding the eggs of salamander (G. M. Smith; 1950). These, and the species of
Characium, Korshikoviella, and Rhopalosolen occurring on plankton animalcules and on
insect larvae may also be instances of symbiosis, but not of a marked nature" (Fritsch,
1935). Protothecaportoricensishas been isolated from the stools of tropical sprue
patients (Mariani, 1942).
Most of the instances of associations of algae with animals mentioned above are
reported froIll Europe, but there are also one or two instances known from the Indian
region. Thus, in Myrionema amboinensisfound by Svedelius (1907) near Galle, Ceylon,
the entire body of the polyps were infested with" :(,oochlorella",the alga being found
in large numbers particularly in the hypostome. The algal cells found in the
endoderm frequently divide into four or eight. Venkataraman (1957) refers to
ECONOMIC IMPORTANCE 51
Chlorellavulgaris as occurring within the freshwater sponge, but he does not refer to the
relationship between the two. Strictly speaking, this species should be considered as
Chlorellaparasitica (Brandt) Beijerinck and that of Svedelius as C. conductrix (Brandt)
Beijerinck.
ECONOMIC IMPORTANCE OF CHLOROCOCCALES
CHLOROCOCCALES AS FOOD OF FISH
It is generally known that a number of aquatic algae form the food offish either
directly or indirectly. Diatoms, filamentous and some planktonic green algae and a
..1. number of blue green algae, exclusive of species forming noxious blooms, are very
appear to be directly utilized as fish food. Apparently, there is also a certain degree of
.selective feeding by some fish' as indicated by the occasional presence of these algae in
good quantity in their guts even when the algae concerned are not dominant in the
water. The reserve food materials i!l these algae, viz. fats and volution in the diatoms,
starch, often accompanied by oil, in ihe green algae; sugars and glycogen in the blue
green algae, and polysaccharides in the Euglenineae, may have some bearing on the
digestibility of the cell contents of these algae since the diatoms (except their shells)'
are the most easily digested and the Euglenineae the least.
Among the green algae other than th~ filamentous forms, t!Ie most important'
~
'" ~an.~ ubiquitous groups in the Indian region are the Volvocales, the Chlorococcales,
and tIu' Desmids. Of these, the Volvocales and some of the desmids found in the
in the guts of fish, but it is the Chlorococcales, which are more often found in fairly
large numbers in ~the guts orfreshwater fish. In the case of the Siamese fish, Tilapia
mossambica,which lias been experimentally introduced at Cutt~ck, and which is
known to be voracious feeder of filamentous algae and some tender macroflora, the
Chlorococcales have been occasionally found in their guts in such large riumbers, that
the_gut ~ontents of this fish have been sometimes quite sufficient to obtain selected
gatherings of various species belonging to the order. According to Prowse (1957),
though the gut contents of Tilapia mossambicastudied at' Malacca were very rich in
diverse Chlorococcales, 'none of them were digested in contrast to the scattered diatoms
which were fully digested. *
However, the Chlorococcales appear to be more important as an indirect source
of food to many fish than as a direct source.. Thus, the enteric cavities of rotifers, co-
pepods, and cladocera are very often filled with various forms of algae, particularly the
Chlorococcales. There is also some experimental evidence to show th~i: some of the
Chlorococcales are utilized as food by these animalcules. In the Limnological
Laboratory of Lund University, Sweden, a culture of Scenedesmus was maintained over
\-a long period, the alga being supplied exclusively with a daily dose of fish meai.,
Another culture exclusively of Daphnia was maintained in the same laboratory by
" feeding the animalcules daily on a certain quantity of Scenedesmus taken from the
~.- .
*The author has also observed faecal pellets of the mullet Mugit corsuta,conSistingalmost exclusively
of undigested
ailla Botryococcusbraunii floating in experimental filter beds at Barrackpore containing both the
Anitth.. /;.h
52 CHLOROCOCCALES ECONOMIC IMPORTANCE 53

cultures. Both Scenedesmusand Daphnia remained perfectly healthy as in nature and
went on multiplying rapidly. * It appears that many other plankton animalcules
could be cultured in a similar manner. When it is remembered that plankton
animalcules are very useful items of food of fish, particularly in the early stages of the
life history of fish (Alikunhi et at., 1955), the potentialities of artificial feeding of fish
and increasing the natural food supply in nursery and rearing ponds could be realized.
Though the utility of phytoplankton as an indirect source of food is very often not
appreciated by many fishery biologists, particularly}n India, on account of the fact that'
the food cpa ins of many of our species of fish haye not been worked out, there is no
doubt that the phytoplankton, of which the Chlorococcales form a very prominent group
in the tropics, playa very important role. It is also known (Schwimmer and Schwim-
Chlorellathan in many other plants; and (6) it is also considered a rich source of vitamins
A, C, and K. B, is also piesent in fairly large proportion, though it might vary with
, the age of cultures. Of these, only vitamin C is likely to be lost considerably in dried
or frozen Chlorella. Witsch (1959) statea that vitamin B, values of young cultures of
, . algae like Chlorella equals that of lemon juice.
Feeding experiments with dry Chlorella powder have shown that animals and
human beings fed on it show a general improvement in health, though it has not been
possible to determine whether it is due to its high protein content or high vitamin
content. However, though the powder has been considered as "food-like" and
" food-satisfying ", its unpleasantly strong smell, general appearance, mildly unpleasant
...

mer, 1955) that several vitamins found in fish' can be ultimately traced t~.the phyto-
plankton on which they feed.
been considered disadvantages.
- -
after.;taste, and a noticeable tightening at 'the ~ck of the throat (" gag-factor ") have
This has been. generally
~ overcome by mixing it with
other items of food like chicken soup so that any unpleasant taste, smell or other dis-
Mention has already b.een made that many Chlorococcales have a cell wall Ii!; advantages are maske<;:l. In Japan, powdered' Chlorella ellipsoidea has been used
which is orqamented with spines, bristles or setae. Apart from the fact that they help successfully after mixing it with green tea or in noodles made of wheat flour, even
these algae to lead a pelagic life, it is also possible that'these armatures offer very
tJ:1ough large qua~tities 'of th~ powder could not be' used o~""a5~~unt of theCiiso'
often useful self-protection against the voraci?us plankton .animalcules. . , -agreeable colour (also-.see Tamlya, 1959). i r~,
,... II" .',.. .. .1;
CHLOROCOCCALESAS A SOURCE OF FOOD AND FEED AND. IN INDUSTRY
. -. [~.o;
Thus, Chlorella has been used more as a food stabilizer (Randhawa;, 1956) than
as a ..c~mplete item of food by itself. * It is possible to raise 20 to 30 time~iIiote
. ..
dried
, ~ ._
\ Investigations on the use of algae as a source of food and feed and of industrial . Chlorellafrom an, acre of ~ater one metre deep than maize from an acr.e ~~ lanci... )
raw materials have been receiving lot of attention all over the world since some tiIl}e. ;c'.
Because of these potentialities, in India the food, value of Chlorella i$<~entl}run~
Many sea weeds, brown and red, and -a few green algae like Ulva- lactuca and .
~nvestig~tion at the Central.Food T?cQnological. Research I~stitq,te at ~so~e. 'T~~~
"u. fasciata, hav€,been used for a long time as a source'of food and feed -or as raw : .
mate;ials for industry, particularly in Japan and China. It is only recently that
fresh-water members of the Chlorococcales, particularly Chlorella, and sometimes
'
~.t "

Scenedumus and Ankistrodesmus, have been thought of for this purpose. Considerable
work has been carried out at the Carnegie Institute of Washington (Burlew, 1953)
and Germany (Witsch, 1959) on mass cultures ofChlorella.and its suitability ~ an alter-
native source of anilI].al feed and of human vegetable food. (Quite recently Russian
scientists are repor~ed to have made Use of Chlorella in space research, .the' effect of
radio-a.ctivity being studied on the alga). , are high (<;omp-arable to that of yeast, it is not, sufficient for extraction as concentrates.
Chlorelfapyrenoidosaand C. vulgaris are.~the two species' st~died in' detail. The
reasons why Chlorella was selected for studies on its utility as food and feed were :
(I) the alga is easily cultured on a mass scale under artificial conditions on account of
its rapid grow1h in mineral solutions; (2) ash analysis of Chlorellashows that its inorganic
contents are more or less on the same level as that of com; (3) the protein, carbotiydrate,
" ,<.
\
IS also a scheme at the Indian AgriculturaL Research InstItute, ~ew Delhi, to study ~,'
the genetic variability of useful algae, including Chlorella.
Regarding the potentialities of the use of Chlorella as raw material'for various
"": .
. ..'.,~""~ indu;tries, *e only one which shows promise" is jts use as a potential source of chlorophyll ~
. -,' for deodOl:ant purposes. This pigment is present in the alga to the ix~nt of 4-6 per
..I. ~ent compared.to O'~ per c<,:ntin alfalfa, whi~~ is now commercially used. for its ~xtrac-
tion (A. W. Fisher In Burlew, 1953). Residual by-products after the extractIOn of
- the pigment canaIso Qt used as)inimal feed: Though the vitamin contents ofChlorella.--
Similarly, ~xtraction of proteins, amino-acids an.d alginic acids also lI].ay not be
economical. Chlorellais also ~nowii to 'have some antibiotic properties since it controls
bacteria to soine 'extent, but work along these lines has yet to be uidertaken in detail.
According to substance.
Gupta and Srivastava (1963) Hydrodictyon reticulatum also produces an
,'J' antibacterial .

and lipide contents and the R-value (which corresponds more or less to the energy
content) in Chlorella.show a great range of variation depending on the environment,
and this range of variation is not exceeded by any other plant. matetial. Further,
it is' possible to induce production of a fairly high content of these under controlled
conditions of cultures; (4) all essential amino-acids are present in Chlorellato the extent
of 42 per cent of its protein content. The amino-acid index for Chlorella is about
62 which is more or less the same as in white flour, pea nut meal, etc.; (5) the pigments
Chlorophyll A, B, Carotene, and ~anthophyll are found in a higher proportion in
*Seen by the author by courtesy of Prof. Sven Thumnark.
- Chlorella has also come in very handy for keeping the air in space vehicles
pure on long interplanetary flights. Recent reports by the New York Times News
~ervice indicate that three members of the life Sciences staff of Aerojet-General
"Corporation, Azusa, California, have developed an 'apparatus and pro<;ess for. the
purpose. The stale air in which the carbon dioxide has been concentfated i~ fed into
}a flood-lit container containihg a mixture of water and nutrient' chemicals and
Chlf!rella. The alga restores oxygen into the space vehicle by its photosynthesis.
It isthe
for alsoastronaut.
expected that the additional alga grown in the process would serve as food
~
· Also, see j\ :ldendum.
 .
CLASSIFICATION AND PHYLOGENY 55

(Protosiphon and Botrydium), (8) Oocystaceae [Eremosphaereae, Chlorelleae-incl.

Tetracoccus (= Wesfella), Micractineae, Oocysteae, Nephrocytieae, Tetraedreae,
Protothecaceae, and the unassigned gen~ra Desmatractum, Acanthosphaera,and Echino-
sphaeridiumJ, (9) Hydrodictyaceae, and (10) Coelastraceae (Scenedesmeae, Sorastreae-
incl. Coelastrum,SorastTum,Dimorphococcus,etc., Selenastreae, and the unassigned genus
CLASSIFICATION AND PHYLOGENY Closteriococcus). The phylogeny of the different families was represented by him as
shown below.
HISTORICAL
SIPHONOCLADIALES' SIPHONALES CHAETOPHORALES
(The classification of -the order Chlorococcales (= Protococcales) has been a very PROTOCOCCALES
difficult one.) A glance at the classifications suggested by various authors over the past
seven decades is sufficient to show the divergent views held from"time to time. This
has been partly due to the advancement in our knowledge of the order with the passage Hyarogastraceae Coelastraceae

~
of time. In 1935, Fritsch pointed out that the classification could not remain natural - Ophiocytiaceae
with the then state of our knowledge. Since that date the order has been in the HYdrO~ictYaceae .

melting pot and even today taxonomists are not agreed on ~fication which is - I
acceptable to all. The classification givenpere is based on views expressed by some of "" '" Oocystaceae
the more important taxono~ists of the order.- '., . ....
Historical resume of the progressivechangesin'.the classification: The order Pr~tococ- - "" protococcacea! (proto!hecaCeaC)
cales, as conceived by Wille (1897), consisted of six families, viz. the Volvoca~eae, I
Tetrasporaceae, Chlorosphaerlfceae, Pleurococcaceae, Protococcaceae, and the Hydro-
dictyaceae. The last family consisted of such genera .as Hydrodictyon, Pediastrum,
Sorastrum, and Coelastrum. The evolu~ionary tendencies ~f these families in relation -
to other orders of the Chlorophyceae were depicted by him as follows:
(RhodOChytria,ceae)
-
\ / -
-
Pleurococcaceae

I (MYl!:~occace~e),
Botryococcaceae

/
C~NJUGATA~
CONFERVOlDEAE
Volvocaceae
_ Hydrodictyaceae Te'nupo,.ocae
~ (H yalovolvocaceae)

~
','
~- _ ~~: Ch!orosphaeraceae .
~ _ -/ ~ PleuroCoccaceae _ Flagellata !'

P-Totoco~caceae
""
_ _ ~-
Tetrasporaceae' _
It was Brunnthaler (1915) who gi\ve the Protococcales the mo!e defined demarca-
tion which has come down to the pr:esent day. His classification of th; Protococcales
was as follows :
""~ >. Ser. I-Zoosporinae Brunnth., 1915,p 59
Fam. Protococcaceae (Protococceae-incl., Ken/rosphaera;Endosphaereae),
Conjugatae---V olvocaceae
PROTOCOCCOlDEAE
- Fam. Characiaceae
Fam. Protosiphonaceae
(incl. Ac/idesmium)
Fam. Hydrodictyaceae
Ser. II~A~tosporinae Brunnth., 1915, p 107
By 1909, he recognized under the order ten families, viz. (1) 'Volvocaceae, Fam. Eremosphaeraceae
Fam. Chlorellaceae (Chlorelleae-inc1. Te/racoccus;-Micractineae)
(2) Tetrasporaceae (inc!. Dictydsphaerium, Dictyocystis, and Chlorosphaera),(3) Botryo- Fam.Oocystaceae Oocysteaej --
coccaceae, (4) Pleurococcaceae (inc!. Gloeotaenium,Acanthococcus,and the Myurococca- Lagerheimieae;
Nephrocytieae (incl. D'sma/rac/um);Tetraedreae;
ceae), (5) Protococcaceae (Endosphaereae, Characieae, Halosphaereae, Botrydio- Faro. Scenedesmaceae Scenedesmeae;
pseae, Chlorothc;cieae, and Rhodochytrieae),. (6) Ophiocytiaceae, (7) Hydrogastraceae ~.,. Selenastreae (incl., Dictyosphaerium and Dimorphococcus)
fl\m. Coe1astracel\e (incl., Soras/rum)
'.
54
 CLASSIFICATION AND PHYLOGENY 57
56 CHLOROCOCCALES

The main features of Brunnthaler's classification was the recognition of two broad Oltmanns (1922) adopted a simple classification recognizing only four families
in his Protococcales, viz. the Protococcaceae (Protococceae, Characieae, Endosphae-
series, the Zoosporinae (reproducing by zoospores) and the Autosporinae (reproducing
reae), the Protosiphonaceae, the Scenedesmaceae (Chlorelleae, Eremosphaereae,
by autospores). Apart from excluding the Volvocaceae, Tetrasporaceae, Botryoco-
Scenedesmeae), and the Hydrodictyaceae. Sorastrumin which zoospores were recorded
ccaceae, Pleurococcaceae, and Ophiocytiaceae of Wille (1909) from the Protococcales
by Probst (1916) was again placed along with Pediastrum, Hydrodictyoll,and Euastropsis
and reassigning some of the genera included in the Tetrasporaceae and Pleurococca-
in its natural place in the Hydrodictyaceae. However, Coelastrum, DictyosPhaerium,
ceae to truly Protococcalean families, he also raised the status of the subfamilies
Raphidium, and allied genera were all placed by him under the Scenedesmeae, and
Characieae, Eremosphaereae, Chlorelleae, and the Scenedesmeae to th~ of families.
Oocystisand allied genera under the Chlorelleae. According to him, the evolutionary
Botr)'dium was excluded from vVille's Hydrogastraceae and this family itself was
tendencies of"the genera under the Protococcaceae are as'follows :
suppressed in favour of the monogeneric family' Protosiphonaceae.
The families Chlorangiaceae, Palmellaceae, Tetraspor~ceae, and Chlorosphae- .. Phyllobium
raceae were separately dealt with by Lemmermann (1915) in th~ same volume under
I
a separate order, the Tetrasporales. Eremosphaera
Till 1915, the accepted name of the order' was Protococcales. Since Protococcus Characium I
(=Pleurococcus) was no longer regarded as a member of the Protococcales (Pascher, Chlorocystis
1915) the term Chlorococcales suggested first by Marchand (1895) and later emended I
Codiolum Chlorochyt rium . .
D lcranoch aete
,

by Pascher (1915) was considered the more appropriate name for the order. However,
it was only from 1927 (West and "Fritsch, 1927) that the name was generally adopted.
According to Papenfuss (1955), the name of the order should read as Chlorococcales
~ - 1,,/
Sykidion
Marchand orth. mut. et emend. Pascher, 1915. . . I
G. S. '::est (1916) adopted the classifications of Wille (1909) and Brunnthaler Chlorococcum (incl. Cystococcum' USW)
(1915) with certain modifications. He recognized under his Protococcales three sub-
orders, viz. the Volvocineae (with three families), the Tetrasporineae (with five Geidel' (1924) suggested the classification of the order into two major groups
on the basis whether the contents of the cell divide gradually or simultaneously at the
families), and the - q,hlorococcineae (with two families). I( the Palmellaceae and .the
Chaetopeltidaceae are excluded from his Tetrasporineae, it corresponds: more or less to time of reprodu~tion. - Howev,er, this has not been considered a satisfactory classifica-
Brunnthaler's Autosporinae, and the Chlorococcineae to the Zoosporinae. Other tion by most authors (G. M. Smith, 1933; Fritsch, 1935).
Korshikov (1926) classified the coccoid algae into two groups, the Vacuolatae
significant changes were the removal of theProtosiphonaceae to the Siphonales and the
(those with' contractile vacuoles) and the Protococ<,:ales (those without contractile
recognition for the first time of a family for the genera Dictyosphaerium,Dictyocystis,- :$
Dimorphococcus,and Westella. He als-o suggested the following evolutionary tendencies
in the order :
.i
. vacuoles), and he depicted the phylogenetic connection among some of the unicellular
forms as follows :
.
:t
SIPHONALES .HIGHER ISOKONTAE
- -------- .~ ~

+
I rT
?

~~ ~r~ ~:;;~~e~~ . ' . ~

~
Macrochloris

«
'

Planosporaceae . Protococcaceae 'p ROT 0 C 0 C C ALE S

- - - - -- --- -
___r , II g
Hydrodictyaceae~.r.:: Palmellaceae 0
~ 0
"'C"°c "';'- 0 Apiococcus
~<% 0c .S <J (, Eo<
5
\
C"? c:.. <J ~...~ 0
').
,
~C"
-g "0
c ~.,
~o
c.;\
"" (,'<,; 0.(,~
CI::
,:I..
VACUOLATAE
(,~
-,;
Hypnomonas

Chlamydomonas
I
Chlamy.domona~ "
Of these, West considered the Tetrasporine tendency the most important
 58 CHLOROCOCCALES

CLASSIFICATION AND PHYLOGENY 59

Printz (1927) followed a classification which was essentially that of Wille (1897,
1909) with some modifications. Apart from the reassignment of a few genera, impor-
Chlorellaceae, Oocystaceae, Selenastraceae, Dictyosphaeriaceae, and the Coelastraceae.
tant changes from Wille's classification were the exclusion of the Botryococcaceae
:1:1I The Selen.astreae was raised to the status of a family for the first time and the Coelas-
(following in this respect Pascher, 1925, who tra~ferred it to the Heterokontae),
Ophiocytiaceae, and the Hydrogastraceae, and the retention of the Protosiphonaceae.
traceae was restricted to Coelastrum,Crueigenia,Scenedesmus, and allied genera.
In 1935, Fritsch adopted a slightly modified classification in which the series
The Chlorosphaeraceae suppressed by Wille in 1909 was also revived. Printz's Tetra-
- Zoosporinaeand Autosporinae were not recognized. According to him, though
poraceae included Myuroeoeeusetc., and the Pleurococcaceae, Desmatraetumand,
Brunnthaler's two series were considered convenient at the time and continued to be
Elakatothrix. The Eremosphaereae, Chlorelleae, Micractinieae, Oocysteae, Gloeo-
followed for about two decades, it was not a natural grouping showing the true affi-
taeniae (inc!. Tetraedron), and the Protothecaceae together formed the' Oocystaceae.
nities of the various families. Fritsch had also done away with all subfamilies (see
The Dictyosphaerieae, Quatematae (= Westella), and Selenastreae were accommodated
along with the Scenedesmeae, CIi1cigenieae, and Coelastreae under the CoelaStraceae. alsoG. M. Smith, 1933). Like West (1916), the Protosiphonaceae was transferred to
the Siphonales. The Chlorosphaeraceae was appended to the order Chlorococcales
The evolutionary tendencies and inter-relationships of his nine families were
depicted by him as follows : , as a special family. The Tetraed.reae included by West and Fritsch (1927) under
'fig:'
'J~r;:..f
'the Ooc y staceae was also transferred to the Chlorellaceae.
!~;: In his classification of the order into eight families Fritsch takes into consideration
~PHONOCLADIALES SIPHONALES " a number of characters, viz. the habitat (free-living, epiphytic, endophytic, etc.);
CHAETOPHORALES l'I ~
. ',habit (solitary or colonial), when colonial, the presence or absence of gelatinized parent
i' '"cell.membranes, connecting threads or mucilaginous pads; the nature and number-of
PROTOCOCCALES I ~~-:;:s. ":!Chloroplasts and the chief mode of reproduction (by autospores, autocolonies or by
I ",
"J 'zooSpores). Though Fritsch did not consider this classification as completely satisfac-
Protosiphonaceae Coelastraceae Pleurococcaceae ~/
\ ~ I' - I ,/'.
/','"
" ,.tory, he was of the opinion that undue emphasis should not be laid on a single
"\' character like autospore or zoospore formation. For, the' same reason he did not agree
"with Korshikov's (1926) classification into the Vacuolatae and the Protococcales, the
Oocystaceae
~ Hydrodictyaceae .
I
Chlorosphaeraceae
,
" :fo~e~ being placed between the Volvocales and the p!otococcales-to accommodate

~
-
'~oid forms which have apparently given up reproduction by swarmei-s, but still

(prth""' )
Chlorococcaceae // "
.:etain contractile vacuoles and sometimes eye-spots.
~; As regards phylogeny, Fritsch (op.e.)stated that, apart from the close affinity
,:to'the Volvocales, the diploid condition in some of the sexually reproducing Chloro-
I ~",," ;f()Ccaleslike Chloroehytnumand probably Apioeoeeus, with tendency for direct germination
(Rhodochytriaceae) ".of~ zygote, is significant in the hypothesis of a possible origin of the Siphonales .
~"
HETEROCONTAE Tetrasporaceae " I ~fromthis order. Also, the coenocytic tendency <.>f
som~ of th~ zoosporic Chlorococcales
CONJUGATAE ~y have led to the evolution of forIns like ProJosiPhon,
which serves as a proto~ypefor
I ~ ,the rest of the Siphonales.
I i,:\-~ Tilden (i935) transferred not only the Protosiphonaceae but also the Hydro-
, (Myurococcaceae) I
Volvocaceae ~ }Iictyaceae and the entire subfamily Chlorochytrieae to the Siphonales, an order unde..
Botryococcaceae I 'i~e subclass" Siphonae" (plant body a single coenocyte), the Chlorochytrieae being
(H yalovolvocaceae) ',accommodated in the family Phyllosiphonaceae. Her Chlorococcales, an order under
1>--'-
------------ :.; ~ the subclass," Uninucleatae ", consisted of three sub-orders, the Volvocineae,
~etrasporineae, and the Chlorococcineae. The Coelastraceae, Oocystaceae, and
FLAGELLATA §c~edesmaceae were placed in the Chlorococcirieae along with some non-Chloro-
~calean families.
West and Fritsch (1921,) adopted, for the first time, the name Chlorococcales for'
the order. Their classification was essentially that of Brunnthaler, the series Zoospor-
_ Smith (1933, 1950) followed a classification which Jas only a variation of the
fystexiufollowed on the continent. In his earlier classification (1933) all subfamilies
inae and Autosporinae being retained. The former consisted of the Chlorococcaceae
~ere dispensed with for the first time, raising the status of some to that of families and
(Chlorococceae-inc!., Charaeium and ChlorosPhaeraand Chlorochytrieae), the Proto-
~ppressing others. He recognized eight families, viz. the Chlorococcaceae, Endos-
siphonaceae, and the Hydrodictyaceae and the latter of the Eremosphaeraceae, eraceae, Characiaceae, Protosiphonaceae, Hydrodictyaceae, Coelastraceae,
taceae, and the Scenedesmaceae. Of these, the prot,: "~ the

60 CHLOROCOqCALEs
Coelastraceae were mono generic. In 1950, he added two more families, the Micracti-
niaceae and the Dictyosphaeriaceae (excl., Westella), the genera belonging to which
were previously accommodated in the Chlorococcaceae and Oocystaceae respectively
Smith (1950) admitted that the Hydrodictyaceae and Scenedesmaceae alone constituted
very natural families, whereas families like the Chlorococcaceae a.nd Oocystaceae were
more or less artificial, the genera under them being grouped together on the basis of
the method of reproduction only.
Prescott (1951) followed the same classification as that of Smith (1933) with
the difference that the Protosiphon'aceae was excluded from the order and the
Botryococcaceae was brought back from the Xanthophyceae.
During the Second World \Var, Korshikov gave a completely new classification
in his Protococcineae, which was posthumously published in 1953. He divided
this large order into three series, viz. Vacuolales, Protococcales, and genera of doubt-
ful systematic position. The series Vacuolales consisted of eight families, viz. the
Hypnomonadaceae, Actinochloridaceae, Palmellopsidaceae, Chlorangiopsidaceae,
Chlorangiaceae, Gloeodendraceae,' Chaetochloridaceae, and the T etrasporaceae ; the
series Protococcales consisted of fourteen families, viz. the. Chlorococcaceae, Characia-
ceae, Chlorosphaeraceae, Borodinellaceae, .Protosiphonaceae, Hormotilaceae, Pal-
mellaceae, Hydrodictyaceae, Oocystaceae, Ankistrodesmaceae, Protococcaceae, Dic-
tyosphaeriaceae, :6otryoc~ccaceae, and the Coelastraceae, and the third series comprised
the genera Elakatothrix, Raphidonema,and Glaucosphaera. ,-
. .This classifiption is jn reality an extension and elabDration of the classification
given by the same author in 1926. In his Protococcineae of 1953",Korshikov not only
included the Vacuolatae, but also a number of palmelloid (both zoosporic and non-
zoosporic) and dendroid genera traditionally included under the Volvocales or Tetras-
porales. Other salient ,features of his classification were the inclusion .of the
Chlorosphaeraceae, Chaetochloridaceae, Botryococcaceae, and the Protococcaceae
within the order; the Protococcaceae being revived to include genera like Protoc()CCUS,
Rl}diococcus,and Coenoc..occU4; the revision of the genus Characium and allied genera;
the creation of a number of new families including the Hypnomonadaceae, Actinochlori-
daceae Palmellopsidaceae; Borodinellaceae, and .the Ankistroaesmaceae; and the
inclusion of the .subfamilies Golenkinioideae and Treubarioideae under the Chloro-
cocc~ceae and the Micractinioideae' under the Coeh\straceae. Apart from reviving the
subfamilies themselves, he revived also their endings in "oideae" which is now
generally accepted as more correct than the endings " ieae " or " eae ".
While recording zoospores in Tet~aedronbitridens Beck-Mannegetta, Starr (1954)
expressed the opinion that Tetraedron be transferred to the family Hydrodictyaceae, as
suggested earlier by Probst (1926). He (1955) also revised the genus Chlorococcum
include Hypnomonas Korshikov, accepting the views of Fritsch and John '(1942) that
their separation into two genera based on the presence or absence of contractile vacuoles
is questionable. 'His comparative study of nine allied genera in the same work has
contributed a great deal to a better understanding of the spherical Chlorococcaceae:
Herndon (1958), after making a detailed study of the Chlorosphaeracean algae
of soil, considered the vegetative division found in the Chlorosphaeraceae of funda-
mental importance.to warrant the creation of a separate order, th~ Chlorosphaerales.
CLASSIFICATION ANI) PHYLOGENY 61
Under this order he recognized two families, viz. the Chlorosphaeraceae (zoosporic)
and the Coccomyxaceae (non-zoosporic). He also suggested the abandoning of the
order Tetrasporales " inasmuch as, in the majority of its genera, cell multiplication
involves the formation of individual cells with completely new walls, the daughter cells
being quite independent of the parent cell membrane". Further, he suggested the
inclusion of all unicellular and colonial Chlorophyceae which are non~motile during
the vegetative phases of their life cycle and which have no capacity for vegetative cell
division ul}der the Chlorococcales and the delimi ting of the Volvocales to' those unicellular
a.nd colonial green algae which are motile during the vegetative phases of theirlife history.
In his Algenkunde,Fott (1959) recognized 12 families under the order Chloroco-
ccales, viz. the Chlorococcaceae, Chlorochytriaceae, Characiaceae, Hydrodictyaceae,
Micractiniaceae, Gloeocystidaceae, Radiococcaceae, Botryococcaceae, Dictyosphaeria-
ceae, Oocystaceae, Scenedesmaceae, and the Protosiphonaceae. Of these, the
Gloeocystidaceae consist of palmelloid zoosporic genera whereas the Radiococcaceae
comprise palmelloid non-zoosporic genera. 'He also recognized the order Tetrasporales
including in it the families Hypnomonadaceae, Chlorangiaceae, Chaetochloridaceae,
Tetrasporaceae, and the Characiosiphonaceae. He did not refer to the Chlorosphaera-
-ceae at all. In 1960, he raised Korshikov's subfamily Treubarioideae to the status
of a family, the Treubariaceae, within the order Chlotococcales.
Bourrelly (1958 a, 1958 b, 1959) gave accounts of members of the order Telra-
sporales under four families, viz. the Chlorangiaceae, Hypnomonadaceae, Palmellaceae,
and Tetrasporaceae. His Chlorangiaceae included CharaciosiphonIyengar and
Characiochloris.Pascher. In 1961(a), he proposed a new classification qf the Chloro-
coccales wherein he' recognized only seven families, viz. the Chlorococcaceae,
iiI
Oocystaceae, Micractiniaceae, Dictyosphaeriaceae, Coelastraceae, Hydrodictyaceae,
and the Coccomyxaceae. Of these, detailed account of only the Chlorococcaceae has
'.;
been published *. Besides Characium and allied genera and Chlorochytriumand allied
genera, this large family of 45 genera include such genera as Tetraedron,Polyedriopsis,
Desmatractum, and Disporopsis. . ,
Thus, the classifications suggested from 1915 onwards, when the order assumed
a definite shape, fall under five broad categories': (I) that of Brunnt~aler (1915) in
which the broad basis-of classification was the mode of repr:.oduc.tion, viz. by- zoospores
or by autospores, (2) that of Geitler (1924) in which the mode of division ofthe cell _
(gradual or simultaneous) at the time of reproduction was alone taken into account,
(3) that of Fritsch (1935) in which a number of characters like habitat, habit, chloro-
plast, mode of reproduction, etc. were all considered together, (4) that of Tilden (1935)
in, which the uninucleate or coenocytic character of the cell was the main criterion,
and (5) that of Korshikov (1926,1953) wherein the presence or absence of contractile
to vacuoles was used for separating.a large, order, the Protococcineae, into two major
series, the Vacuolales and the Protococcales. Other authors, though mainly foJlowing
modifications of Wille (1897, 1909), Brunnthaler (1915), Fritsch (1935) or Korshikov
. *In the other families subsequently published (Bull. d. Microsc. Appl. (2), 13 (5): 113-43, 1963;
.bid, 13 (6):155-86, 1963), interesting features are the inclusion of Botryococcus,Botryosphaera, Actinodesmium
.:.(? Actidesmium), Dictyochlorella, etc., under the Dictyosphaeriaceae, Paradoxia, Dicellula, Radiococcus,
Actinastrum, Gloeoactinium, etc., under the Coelastraceae and Elakato/hrix under the Coccomyxaceae.
 M CJtLOROCOCCALE~
(1953) have also contributed much, in their own way, in explaining the phylogeny of
the different families and genera under the order.
Within the past two and a half decades some important additions to our knowledge
of the order have also been made. Iyengar (1936) discovered the unique genus
Characiosiphonwith a protocoenocytic thallus, for which he proposed a new family, the
Characiosiphonaceae. According to him, it has been probably derived from some
unicellular ancestor resembling Characiochloris. Korshikov (1937) recorded oogamous
reproduction in Golenkinia longispina, G. solitaria, and Micractinium pusillum, and on this
basis he (1953) separated these species of Golenkinia into a new genus Golenkiniopsis.
Oogamy has since been recorded in Dictyosphaeriumindicum Iyengar et Ramanathan
(1940), in Golenkiniopsis minutissima (Iyengar et Balakr.) (=Golenkinia minutissima
Iyeng. et Balakr.) and in a new genus OocystaeniumGonzalves et Mehra (Oocystaceae),
having numerous chloroplasts. Iyengar (1962) has also recorded a new tlendroid
attached genus, Dendrocystis, belonging to the Lagerheimioideae.
Zoospore formation has been recorded in a number of genera and species which
were known to reproduce either by autospores or by autocolonies or in which re-
production was not known at all. Thus, zoospores' were recorded iri AcanthosPhaera
(G. M. Smith, 1950), DictyosPhaerium ierrestre (Fritsch and John, 1942), Tetraedron
bitridens (Starr, 1954), PlanktosPhaetia spp. (Starr, 1954 a; Herndon, 1958 a) and Poly-
edriopsis sPinulosa (Korshikov, 1953). It .is on the basis of zoospore formation in
AeanthosPhqeraand possibly in allied genera that Smith (1950) has also raised the status of
the Micractineae to that of a family_ KorshiKov (1953) has also recorded eye-spots
in the daughter cells, of Trochiscia reticltlaris (Fig. 22e), which genus he considers as
allied to Golenkinia. According to Starr (1954 a), the discovery of zoospores in Plank-
tosphaerlamakes it necessary to transfer this genus from the Oocystaceae, where Smith
originally placed it, to the spherical Chlorococcaceae. The record of zoospOres in
Schroederia(see Korshikov, 1953) also makes it possible t() keep this genus by the side of
Characium. Schroederia-like algae with a bifurcate stalk and reproducing by zoospores
a
have been 'placed in new genus Ankyra'by Fott (1957);
, K. P. Singh (1960) recorded macro- and miCro-autospores in a new spec!es of_.
. Oocystzs from Kumaop Hills, India. - -
01]._the _taxonomic side; though ,a number of new genera (particularly by Pascher,
e- Korshikoy and others) have-been added to the Chlorococcales during the past 25,years,
life history studies have definitely proved that a' few genera which were originally
included under the Chlorococcales do not belong to this order at-all or sometimes even
to the algae. Thus, Placosphaera Dangeard has been shown (Thompson, 1956) to
be a stage in the life history of Schizochlamys (Volvocales or Tetrasporales). Codiolum
gregarium A. Braun and C. petrocelidis Kuckuck have been shown (Jorde, 1933; Fan,
1959) to be the sporophytes of Urospora mirabilis Arechoug and SpongomorPhacoalita
(Ruprecht) Collins respectively. Some other species are also under suspicion.
Instances of wrong references {)f other plant material to the Chlorococcales are
"Phytoniorula" Kofoid (19i4), "Thamniastrum" Reinsch (1867) and some speci~
of" Cerasterias" (Copeland, 1937; Taft, 1945: G. M. Smith, 1950). These" genera"
or "species" become naturally excluded from the Chlorococcales or suppressed
altogether as the case may be;.
CLASSIFICATION AND PHYLOGENY b3
As in the case of a number of species of Characium (Printz, 1927) which have been
found to belong to Characiopsis (Xanthophyceae) a number of species of Tetraedron
are under suspicion because of the existence of " Tetraedron-like" stages in the life
cycles of Pediastrum, Hydrodictyon, and Oocystis (West and Fritsch, 1927 and Fritsch,
1935). Some of the described forms could also belong to the resting stages of other
algae. Similarly," Trochiscia-,stages" are known to occur in the life history of
Chlorococcum (seeFritsch and John, 1942). According to West and Fritsch (op. cit.),
" the recording of such forms, even as new species. " . . . . . ,admits of their temporary
classification until they are assigned elsewhere".
Since pyrenoids and starch have not been demonstrated in a number of species
of Tetraedronand since the chromatophores are also several, Skuja (1948, 1949) refers
such species to new genera under the Xanthophyceae. Bourrelly (1951), Korshikov
(1953), Fott (1959), and Fott and Komarek (1960) also do not recognize all the known
species of Tetraedron (also see under Tetraedron on plIO). More detailed observa-
tions on these doubtful species appear to be warranted to make sure whether there are
parallel forms of a}most identical structure belonging to the Chlorococcales as well
~ the Xanthophyceae. - ',._
DichotomococcusKorshikov, formerly regarded -as one of the Dictyosphaeriaceae,
is also now considered as a member of the Xanthophyceae (Fott and Komarek, 1960).
Exactly the reverse has been the case with Botryococcus. Considered as a member of
the Chlorophyceae till 1925 and then transferred by Pascher to .the Heterokontae, the
genus is now 'more or less definitely broug~t back to the Chlorococcales on evidence
pUt forward' by Blackburn (1936), Belcher andFogg (1955) and several others (see
under Botryococcus).
-' The advent of the electron microscopehas also contributed to a better knowledge
of the internal Structure of a number of algae including the Chloro.coccales (Desikachary,
1957, [959). This has been particularly the case with regard to the cell wall, chloro-
. plast, and pyrenoid. ,
, Though these additions to our knowledge during the past quarter of-a century
,.have made it necessary for a number of authors t!' revise the classification of the
Chlorococcales,- even today it cannot be said that a completely satisfactory classification
is available because the affinities of many genera are still imperfectly known. Several
- aUthors still find it convenient _ to separate predqminantly zoosporic families from
non-zoosporic ones in spite of the fact that it is becoming more and more evident- that
'a number of genera which were formel-ly supposed to be autosporic do reproduce
by zoospores under certain favourable conditions. It is, therefore, quite possible
that the two types of reproduction known to occur in genera like Desmatractum, Octo-
gonfella,PlanktosPhaeria,and Tetraedronmight exist in a number of other genera, but one
or other of the methods has apparently become more suitable'in the particular environ-
.Dlentin which the alga lives. Sometimes, as in Actidesmiurnand Marthea,a zoospore-
;"'..like stage is passed through before the autospore or auto-colony is formed. The
,: :Occasional formation of autospores in ,Characium terrestris (Kanthamma, 1940) and the
if..formation of zoospores in DictyosPhaeriumterrestreare also revealing since the formation
,of ZOOsporesand auto-colonies are more common in the respective genera. Though
~the aforesaid instances might help in tracing the evolutionary tendencies among the
 CLASSIFICATION AND PHYLOGENY 65
6"~ CHLOROCOCCALES

various families and genera, it does not necessarily follow that all gencra showing
(see also,
reproduction by zoospores (or by autospores) should be grouped together
Fri tsch, 1935).
The very recent discovery of Trainor (1963) that Scelledesmus obliquusproduces
biflagellate swarmers with a parietal chloroplast, without any pyrenoid and apparently
without a cell wall, in basal media from which ammonium nitrate has been excluded
and his subsequent finding (Trainor, ill press, from personal communication) that these
swarmers are re'ally gametes which are produced by individuals of different strains
(Heterothallic) and which elump in pairs at low temperatures (15°C) in media low in
nitrogen to form quadriflagellatt:: zygotes, are extremely interesting since it changes 1. Unicellular or in loose aggregates, rarely colonial; free-living, rarely in association with other plant.
or animals; cells usually spherical and with or without contractile vacuoles; chloroplasts onc to
our present concept of the genus and necessitates a reconsideration of its relationships several; reproduction by zoospores or gametes Fam. Chlorococwceae
and classification of this genus. Equally interesting is the record of motility in Coelastrum
2. Macroscopic clusters of club-shaped coenocytes without Cro.5 wal," between protoplasts; cells
(Trainor and Burg, 1964-not scenby the author). According to Trainor (l.c.), the with several contractile vacuoles; chloroplast stel1ate; reproduction by zoospores or gametes.. .. ..
feasibility of maintaining two families of colonial or coenobial forms like the Hydro- Fam. Characiolipho1!aceae
dictyaceae and Coelastraceae (the latter including Scenedesmus and its allies) is q~eg... 3. Unicellular (rarely colonial); usually attached, rarely free-living; cells usually elongate and
tionablc, as earlier hinted by Fritsch (1935). Since the above information was sometimes with contractile vacuoles; chloroplast parietal and laminate; reproduction by zoospores
recei,,~ed by the ~uthor after sending-the present work to the press, no changes in the or gametes.. . . . . Fam. Characiaceae
classification is attempted. It is sufficient to mention here that in three of the pre- 4-. Unicellular; usually endophytic, rarely epiphytic or free-living; cells large and irregularly
dominantly autosporic families, viz. the Oocystaceae, Dictyosphaeriaceae, and the thickened; chloro\-:...ls axial and massive; reproduction usually by numerous zoospores
orgametes.
..... . Fam. Chlorochylriaceae
(Endosphaeraceae)
Scenedesmaceae and possibly in the Coelastraceae, motile sexual cells have now been
established at least in some typical genera. Unicellular or colonial; free-living; cells more or less spherical; ccll wall with ridges, warts,
spines, bristles or setae; chloroplast single, parietal and cup-shaped or several and disc-shaped;
According to Bendix (1964), who studied the" phenotypic variability in certain reprod~ction usually oy autospores, rarely by zoospores or oogamous gametes.. . . . .. .
Fam. Micra£lilliaceae
ChlOrellapyre/loidosQstrains", his observation that Chl;rellaproduces motile cells which
might be gametes, as observed earlier by Pearsall and Loose (1937), and the record of 6. Usually unicellular; free-living or rarely attached; cells spherical, ellipsoid, fusiform or tetrahedric
biflagellate swarmers in Scenedesmus obliquusby Trainor (t.c.) support the possibility that 'with projecting ridges at margins; often with setae from angles or ends; cell membrane differentiated
into two layers, an inner thin firm one and an outer layer made up of two or more parts; reproduc-
Chlorella,an alga presently considered to be asexual, is in fact sexual~ Whether Chlorella tion by zoospores (or aplanospores) or autospor~s.. Fam. Treubariaceae
reproduces sexually or not, he states that our present concept of its iife cycle must be
7. Unicellular or in regular free-living colonies; cells tetrahedral or polygonal, sometimcs nearly
revised after obtaining more information. spnerical; chloroplast parietal-and laminate; reproduction by zoospores (which usually form auto-
The classification adopted in this account is, one in which the vacu~late coccoid colonies) or by autos~ores or gametes Fam. Hydrodic£vaceae
forms are ;etained within the order, whereas Chlorosphaera and allied g:enera which (a) In regular free-living colonies (net-like, flat discs or spherical); celis-polygonal, sometimes
posSess definite vegetative cell division and which are placed by Herndon (1958) nearly spherical; reproduction by zoospores which form auto-colonies or by gametes......
, Subfam. Hydrodictyoideae
under the Chlorosphaerales are excluded from the order. - This- is due- to the fact
that_ in genera like ChlorococcllTII,Actinochloris,Spongiococcus,and Bicuspidella there are (b) Usually unicellular; free-living; cells usually tetrahedric, often with spines from angles,
spe~es with or without contractile vacuoles. As regards Chlorosphaeraand its allies, rarely semilunar; reproduction by autospores, rarely by zoospores.. -
Subfam. Telraedrolloideae
not only they an: still imperfectly known but also no member has so far been reported
8. Unicellular or colonial; when colonial, usually within a gelatinous envelope; free-living, rarely
from the Indian region. Since the inclusion of truly palmelloid forms within the symbiotic; cells of different shapes; chloroplasts one to numerous; reproduction by autospores,
Chlorococcales is a llluch larger question involving discussion of a part of the Volvo- rarely by oogamousgametes.. . . . . Fam:Oocyslaceae
calc:s, they are also left out of the order as defined here. .The proposal by Starr (1954) (a) Usually solitary and planktonic, rarely in attached dendroid"colonies; cells spherical to elli-
to mclude Tetraedron under the Hydrodictyaceae (as originally suggested by 'Probst psoid; cell wall with bristles (rarely verrucose); chloroplasts one or more and parietal; repro-
duction by autospores Subfam. Lagerheimioideae
(1926) on the basis of his observation of zoospores in T. minimum)appears to be further
streng~en.ed by the record of zoospores by Korshikov (1953) in the allied genus (b) Usually solitary; free-iiving or symbiotic; cells usually spherical or ellipsoid; cell wa1l5mooth
or sometimes ornamented; chloroplasts one or rarely more; reproduction by autospores......
Poge~su. Tetraedron and allied genera are, therefore, included here in the Subfam. Chlorelloideae
Hydrodictyac~e as a separate subfamily. The families Radiococcaceae (Fott, 1959)
(c) Unicellular; free-living; cells usually large, spherical, ellipsoia..'br naviculoid; with many
and Tr~u~ceae (¥ott, .1960) are retained along with twelve other more well disc-shaped chloroplasts; reproduction by autospores or aplanoSpores, rarely by oogamous
known families to make a ~tal of. 14 families within the order. Following Korshikov gametes Subfam. Eremosphaerdideae