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Marine Biology 37, 377-387 (1976)

9 by Springer-Verlag 1976

The Concept of Light Intensity Adaptation in Marine


Phytoplankton: Some Experiments with Phaeodactylum
tricomutum
J. Beardall I and I. Morris 2

1 Department of Botany and Microbiology, University College London; London, England, and
2 Bigelow Laboratory for Ocean Sciences (A Division of Northeastern Research Foundation);
West Boothbay Harbor, Maine, USA

Abstract

The historical background on adaptation of algae to various light intensities is


analysed. It is argued that there is little evidence to suggest that previous
growth at low light intensities enhances the ability of an alga to utilize these
low light levels. Rather, the published evidence suggests that the most general
response to growth at sub-optimal light intensities is a reduced ability to uti-
lize saturating levels. The present experiments with Phaeodactylum tricornutum Bohlin
have tested this concept of light intensity adaptation. Changing photosynthetic
abilities during batch growth depended on the light intensity used for growth and
these changes affected interpretations of the data. When m e a s u r e m e n t s were m a d e
at a single time towards the end of the growth curve, cells grown at lower light
intensities appeared to p h o t o s y n t h e s i z e (at all intensities) better than did
those grown at higher light levels. When the changes during b a t c h growth were
taken into account, or when the alga was grown in turbidostat cultures, a dif-
ferent picture was obtained. Growth at reduced light intensities was accompanied
by (a) increased chlorophyll content, (b) decreased rate of light-saturated photo-
synthesis expressed on a chlorophyll, cell number or cell protein basis, and (c)
decreased activity of RuDP carboxylase. One result suggested that growth at a sub-
optimal light intensity did enhance the ability to utilize lower light levels. The
l i g h t - s a t u r a t i o n curve of cells grown in batch culture at 0.7 klux showed higher
slopes at the low light intensities than did those grown at 12 klux. This was most
marked w h e n p h o t o s y n t h e s i s was expressed per cell, but was also apparent when it
was put on a per chlorophyll basis.

Introduction tures does not enhance their ability to


utilize such lower temperatures and that
In 1966, Yentsch and Lee cited the re- the earlier evidence indicating that al-
mark of D.M.S. W a t s o n (addressing the gae do gain such an ability resulted
B r i t i s h A s s o c i a t i o n in 1929): "There is from changing p h o t o s y n t h e t i c abilities
no branch of zoology in w h i c h assumption during growth in b a t c h c~Iture. Rather,
has played a greater part or evidence a it appears that the general response of
lesser part than in the study of adapta- algae to growth at lower temperatures
tion." Almost half a century later, the is a loss of ability to utilize the
same sentiment can be expressed over con- higher optimal temperatures.
tinued attempts to define the ways in In this paper, we present studies of
which p h y t o p l a n k t o n are "adapted" to the the response of Phaeodactylum tricornutum
dominant environmental factors such as to growth at various light intensities
light, temperature and nutrient concen- suggesting that "adaptation" to light
tration. intensities be considered as comparable
Recent w o r k from our laboratory (Mor- to the recently m o d i f i e d view of temper-
ris and Glover, 1974) questioned the ature adaptation. That is, the data sug-
generally accepted hypothesis for temper- gest that a major response of P. trieornu-
ature adaptation. Data from a number of tum to growth at sub-optimal light in-
different m a r i n e algae suggested that tensities is a loss of ability to use
previous growth at sub=optimal tempera- saturating levels. However, some evi-
378 J. Beardall and I. Morris: Light Adaptation in Phytoplankton

4
i a from d e e p e r w a t e r are b e t t e r able to uti-
lize low light i n t e n s i t i e s . A c l o s e r ex-

o 2~ ,. a m i n a t i o n of the o r i g i n a l data (Fig. la)


i l l u s t r a t e s the p o i n t that the r e d u c t i o n
in I k r e s u l t s s i m p l y from lower r a t e s of
~, 2 l i g h t - s a t u r a t e d p h o t o s y n t h e s i s (Pmax)
E and not f r o m any i n c r e a s e d a b i l i t y at
lower light i n t e n s i t i e s . C o m p a r a b l e data
have b e e n p r e s e n t e d by I c h i m u r a et al.
c~
E (1962) and A r u g a and I c h i m u r a (1968).
n" I i ~ i I , i J , ) i r i , I Later, Y e n t s c h and Lee (1966) r e - e x -
a m i n e d the c o n c e p t of light i n t e n s i t y
b a d a p t a t i o n a m o n g s t p h y t o p l a n k t o n , and

so
I /;//_x e m p h a s i z e d the fact that r e d u c e d I k val-
ues b r o u g h t a b o u t by d e c r e a s e d v a l u e s of
Pmax d i d not m e a n an e n h a n c e d a b i l i t y
to u t i l i z e lower light i n t e n s i t i e s .
T h e s e w o r k e r s also s u g g e s t e d that such
c h a n g e s in Pmax and I k c o u l d be i n d u c e d
by m e t h o d s o t h e r than g r o w t h at r e d u c e d
light intensity; that they w e r e best
t h o u g h t of as r e s p o n s e s to an " i n f e r i o r
0 , , , , I , , , J I e n v i r o n m e n t " and not as s p e c i f i c a d a p t a -
F 5000 I0,000 15,000 tions to s u b - o p t i m a l light i n t e n s i t i e s .
Lux S t e e m a n N i e l s e n and J ~ r g e n s e n (1968)
c a r r i e d the d i s c u s s i o n f u r t h e r by re-
Fig. I Light saturation curves for photosynthesis p o r t i n g the r e s u l t s of some e x p e r i m e n t s
of Arctic summer plankton from three depths, i: with ~lorella pyrenoidosa. These w o r k e r s
Surface; 2 : 2 7 m; 3 : 5 0 m. (a) Gross photosyn- e m p h a s i z e d the d i f f e r e n c e b e t w e e n ex-
thesis per unit chlorophyll; (b) normalized p r e s s i n g p h o t o s y n t h e t i c r a t e s per cell
curves (Pmax is put at iOO). (From Steeman Niel- and per c h l o r o p h y l l . In this alga, it
sen and Hansen, 1959) was argued, a d a p t a t i o n to r e d u c e d light
i n t e n s i t y was a c h i e v e d m a i n l y t h r o u g h
i n c r e a s i n g the c e l l u l a r c o n t e n t of chlo-
rophyll. As a r e s u l t of this, the rate
d e n c e s u g g e s t s an i m p r o v e d u t i l i z a t i o n of p h o t o s y n t h e s i s per cell was g r e a t e r
of low light i n t e n s i t i e s by cells pre- in cells p r e v i o u s l y g r o w n at lower light
v i o u s l y g r o w n at r e d u c e d light levels. i n t e n s i t i e s . T h a t is, g r o w t h at sub-
An a p p r e c i a t i o n of the p o s s i b l e gen- o p t i m a l light i n t e n s i t i e s did not en-
eral r e l e v a n c e of t h e s e o b s e r v a t i o n s h a n c e the e f f i c i e n c y w i t h w h i c h c h l o r o -
d e p e n d s on an u n d e r s t a n d i n g of the some- p h y l l u t i l i z e d r e d u c e d i n t e n s i t i e s , but
what c o n f u s e d l i t e r a t u r e w h i c h has ap- it did i n c r e a s e the rate of p h o t o s y n -
p e a r e d on light a d a p t a t i o n of p h y t o p l a n k - thesis on a c e l l u l a r basis by i n c r e a s i n g
ton. For this r e a s o n we p r e s e n t the his- the c h l o r o p h y l l content. This p o i n t is
tory of the s u b j e c t in m o r e d e t a i l than s u m m a r i z e d in Fig. 2, taken from Fig. 2
is usual. In 1957, T a l l i n g i n t r o d u c e d of S t e e m a n N i e l s e n and J ~ r g e n s e n (1968).
the c o n c e p t of I k , the light i n t e n s i t y T h e s e o b s e r v a t i o n s of S t e e m a n N i e l s e n
at w h i c h p h o t o s y n t h e s i s r e a c h e s s a t u r a - and J ~ r g e n s e n (1968) w e r e the first ones
tion. In 1959, S t e e m a n N i e l s e n and suggesting that so-called "adaptation"
H a n s e n e m p h a s i z e d this p a r a m e t e r , I k , to r e d u c e d light i n t e n s i t y i n v o l v e d an
in their i n t e r p r e t a t i o n of light inten- i n c r e a s e d a b i l i t y to use such low l i g h t
s i t y / p h o t o s y n t h e s i s c u r v e s for n a t u r a l levels. The same a u t h o r s r e c o g n i z e d ,
p o p u l a t i o n s of p h y t o p l a n k t o n . The cen- however, that this p h e n o m e n o n was not
tral p o i n t w h i c h S t e e m a n N i e l s e n and universal amongst algae pointing partic-
H a n s e n m a d e can be i l l u s t r a t e d by Fig. I u l a r l y to o b s e r v a t i o n s w h i c h s u g g e s t e d
(taken from Figs. I and 2 of S t e e m a n that " a d a p t a t i o n " of C y c l o t e l l a m e n e g h i n i a n a
N i e l s e n and Hansen, 1959). By n o r m a l - to low light i n t e n s i t i e s w a s not c a u s e d
izing the data (Pmax is put at 1OO) from by c h a n g i n g c h l o r o p h y l l content. Rather,
Fig. la, e m p h a s i s is p l a c e d on the in- in this d i a t o m the m a j o r c h a n g e was in
t e n s i t y at w h i c h p h o t o s y n t h e s i s satu- the r a t e of l i g h t - s a t u r a t e d p h o t o s y n -
rates, and this is c o n s i d e r e d to be thesis per cell, s u g g e s t i n g a c h a n g e in
the d o m i n a n t i n f l u e n c e of the p r e v i o u s the level of e n z y m e s i n v o l v e d in the
light h i s t o r y - i.e., the d e p t h from dark r e a c t i o n s of p h o t o s y n t h e s i s .
w h i c h the p h y t o p l a n k t o n s a m p l e is taken. T h e r e are a n u m b e r of u n c e r t a i n t i e s
S u p e r f i c i a l l y , the n o r m a l i z e d curve in the i n t e r p r e t a t i o n s of the data of
(Fig. Ib) s u g g e s t s that p h y t o p l a n k t o n S t e e m a n N i e l s e n and J ~ r g e n s e n (1968).
J. Beardall and I. Morris: Light Adaptation in Phytoplankton 379

,% -~ Klux 0

E ,% I Klux

0.32 ,ux

0 5' I'0 I'5 2i0 25,

I Klux
X ~

X
f
O9
--I
X
/
_1
I,I
x 3 Klux 9 ,
0", 5 /.e--
0
/ * n 0,32 Klux

o 3

//7 K, ...------Z%

r ~...- J I l I I
0 5 10 15 20 25 30
LIGHT INTENSITY

Fig. 2. Chlorel!a pyrenoidosa. Light saturation curves for photosynthesis by algae grown at 4 dif-
ferent light intensities. Photosynthesis expressed on per unit chlorophyll (a) and cell-number (b)
basis. (From Steeman-Nielsen and J~rgensen, 1968)

F i r s t l y , the d i f f e r e n c e b e t w e e n Chlorella c u r r e d at the lowest light i n t e n s i t i e s .


pyrenoidosa and Cyclotella meneghiniana m i g h t Finally, it is not clear w h e t h e r the en-
not be as m a r k e d as the w o r k e r s suggest. h a n c e d c h l o r o p h y l l c o n t e n t w a s an "adap-
For example, cells of Ch!orella pyrenoidosa tation" in the sense that it i n c r e a s e d
s h o w e d m o s t s i g n i f i c a n t i n c r e a s e s in the g r o w t h rate of Chlorella pyrenoidosa at
c h l o r o p h y l l l e v e l s o n l y w h e n g r o w t h oc- low light i n t e n s i t i e s .
c u r r e d at the lowest light i n t e n s i t i e s One f u r t h e r o b s e r v a t i o n of S t e e m a n
(less t h a n 3 klux) w h e r e a s the e x p e r i - N i e l s e n and J ~ r g e n s e n (1968) is w o r t h
m e n t s w i t h Cyclotella menighiniana did not comment. W h e n e x p r e s s e d per u n i t cell
c o n s i d e r light i n t e n s i t i e s less t h a n 3 number, g r o w t h at low i n t e n s i t i e s (ex-
klux. Also, the i n c r e a s e d c h l o r o p h y l l cept the lowest, 0.32 klux) not o n l y in-
c o n t e n t i n d u c e d by g r o w t h of Chlorella c r e a s e d p h o t o s y n t h e s i s at low light lev-
pyrenoidosa at l o w light i n t e n s i t i e s w a s els, it also i n c r e a s e d Pmax (Fig. 2b) .
not the only change; o t h e r c h a n g e s re- In that sense, t h e r e f o r e , the c h a n g e s
s u l t i n g in an i n c r e a s e d cell size oc- b r o u g h t about by g r o w t h at s u b - o p t i m a l
380 J. Beardall and I. Morris: Light Adaptation in Phytoplankton

light i n t e n s i t i e s w e r e not s p e c i f i c to m e d i u m c o n t a i n i n g n i t r a t e as the sole


the light r e a c t i o n of p h o t o s y n t h e s i s source of n i t r o g e n was used.
(i.e., to the u t i l i z a t i o n of low light S t e a d y states w e r e m a i n t a i n e d for at
levels), but w e r e g e n e r a l for the e n t i r e least 3 g e n e r a t i o n times b e f o r e h a r v e s t -
photosynthetic machinery. ing the culture.
In this p a p e r we q u e s t i o n w h e t h e r
g r o w t h of Phaeodactylum tricornutum at re-
d u c e d light i n t e n s i t i e s i m p r o v e s the Measurement of Photosynthetic Carbon Dioxide
p h o t o s y n t h e t i c c a p a c i t y at any or all Assimilation
light i n t e n s i t i e s . In d o i n g this, we
have r e c o g n i z e d the need to c o n s i d e r Cells w e r e h a r v e s t e d by c e n t r i f u g a t i o n
c h a n g e s w h i c h occur d u r i n g g r o w t h of the at 6000 rpm and s u s p e n d e d in fresh "f-1"
alga in b a t c h culture, to u t i l i z e con- m e d i u m at a d e n s i t y of 2 x 105 cells/ml.
t i n u o u s c u l t u r e s o p e r a t i n g as light- For t u r b i d o s t a t g r o w n cultures, m e d i u m
l i m i t e d t u r b i d o s t a t s , and to m e a s u r e ac- c o n t a i n i n g n i t r a t e only was used. P h o t o -
t i v i t i e s of p h o t o s y n t h e t i c e n z y m e s as s y n t h e s i s was then m e a s u r e d by the m e t h -
well as w h o l e - c e l l rates of p h o t o s y n - od d e s c r i b e d by B e a r d a l l and M o r r i s
thesis. (1975) u n d e r an air a t m o s p h e r e . L i g h t
i n t e n s i t y was v a r i e d by a d j u s t i n g the
d i s t a n c e b e t w e e n light s o u r c e and the
Materials and Methods cell s u s p e n s i o n a n d / o r c o v e r i n g the ves-
sel w i t h muslin.

Growth of Phaeodactylum tricornutum in Batch


Culture Measurement of Enzyme Acitivities in Cell-Free
Extracts
Phaeodactylum tricornutum B o h l i n (obtained
from the C a m b r i d g e C u l t u r e C o l l e c t i o n ) Cell e x t r a c t s w e r e p r e p a r e d and RuDP
was g r o w n in the "f-1" m e d i u m of Guil- c a r b o x y l a s e [E.C. 4. 1.1. 39; 3 - p h o s p h o -
lard and R y t h e r (1962) u n d e r c o n t i n u o u s D - g l y c e r a t e c a r b o x y l a s e (dimerising)]
i l l u m i n a t i o n as d e s c r i b e d by G l o v e r et al. m e a s u r e d as d e s c r i b e d by B e a r d a l l and
(1975). L i g h t i n t e n s i t y was v a r i e d by M o r r i s (1975) M a l a t e d e h y d r o g e n a s e (E.C.
c o v e r i n g the c u l t u r e v e s s e l s w i t h layers 1.1.1. 37 C l-malate: N A D o x i d o r e d u c u
of muslin. For all e x p e r i m e n t s , c u l t u r e s tase) was a s s a y e d by the m e t h o d of M o r -
were i n o c u l a t e d from s u s p e n s i o n s m a i n - ris and S y r e t t (1965) in a U n i c a m S P
tained at the a p p r o p r i a t e light inten- 18OO s p e c t r o p h o t o m e t e r u s i n g a s y s t e m
sity for 2 to 3 days. G r o w t h was fol- c o n t a i n i n g : 0.6 ml O . 1 M T r i s - H C l buffer,
lowed by m e a s u r i n g cell n u m b e r s (with a pH 8.6, O.1 ml of 2.0 m M NADH, O.1 ml of
h a e m a c y t o m e t e r ) twice daily. O . 1 M M g C I 2 and O.1 ml of extract. The
blank cuvette contained Tris-HCl buffer
in p l a c e of NADH. O.1 ml of 10 m M o x a l o -
Growth of Phaeodactylum tricornutum in Tur- a c e t a t e was added to each c u v e t t e and
bidostat Culture the rate of c h a n g e in a b s o r b a n c e at 340
nm d e t e r m i n e d .
Turbidostat c o n t i n u o u s - f l o w c u l t u r e s of
Phaeodactylum tricornutum w e r e m a i n t a i n e d in
a 10 1 glass v e s s e l w i t h a w o r k i n g vol- Measurement of Chlorophyl] a and Carotenoids
ume of 8 I, s t i r r e d w i t h a s t a i n l e s s -
steel b a f f l e at 200 rpm. The g r o w t h ves- C h l o r o p h y l l a was d e t e r m i n e d by the m e t h -
sel was i l l u m i n a t e d by two banks of 5 od of L o r e n z e n (1967). B e t w e e n 50 and
"warm white" f l u o r e s c e n t tubes g i v i n g a 1OO ml of cell s u s p e n s i o n w e r e f i l t e r e d
light i n t e n s i t y at the s u r f a c e of 11 t h r o u g h g l a s s - f i b e r discs, m o i s t e n e d
klux. w i t h a drop of s a t u r a t e d m a g n e s i u m car-
A s e l e n i u m p h o t o c e l l a t t a c h e d to the b o n a t e solution. The cells w e r e h o m o g -
s u r f a c e of the g r o w t h v e s s e l m e a s u r e d e n i z e d for I m i n in 3 ml 90% (v/v) ace-
light t r a n s m i t t e d t h r o u g h the c u l t u r e tone. A f t e r w a s h i n g w i t h I ml 90% ace-
from the g r o w t h lights. The v o l t a g e gen- tone, the b r o k e n s u s p e n s i o n was c e n t r i -
e r a t e d was p r o p o r t i o n a l to the cell den- fuged and the a b s o r p t i o n of the super-
sity of the c u l t u r e and the a m p l i f i e d n a t a n t m e a s u r e d at 665 nm w i t h a U n i c a m
signal o p e r a t e d a r e l a y c o n t r o l l i n g a S P 1800, b e f o r e and after a d d i n g a d r o p
p e r i s t a l t i c pump. At p r e - s e l e c t e d cell of I N HCI. The c o n c e n t r a t i o n of c h l o r o -
d e n s i t i e s the pump was o p e r a t e d and p h y l l a was then c a l c u l a t e d u s i n g the
fresh m e d i u m t r a n s f e r r e d from a reser- e q u a t i o n of L o r e n z e n (1967). C a r o t e n o i d
voir to the g r o w t h vessel. C o n s t a n t vol- c o n c e n t r a t i o n s w e r e d e t e r m i n e d u s i n g the
ume was a c h i e v e d by m e a n s of an o v e r f l o w same s u p e r n a t a n t b e f o r e acid t r e a t m e n t .
system. For t u r b i d o s t a t cultures, "f-1" The a b s o r b a n c e at 480 and 750 nm was
J. Beardall and I. Morris: Light Adaptation in Phytoplankton 381

//
measured, and c o n c e n t r a t i o n calculated 0.06
using the equations of Parsons and
Strickland (1963).
0,05
L

=" 0.04
Results kd
F-
n-"

T
f-
0.0~
Effect of Light Intensity on Growth Rate
0
rr
o
It is important that the response of 0.02
photosynthesis to light intensity be
related to growth. That is, it is neces-
.y
sary to specify the relationship be-
tween any "adaptation" identified in
photosynthesis measurements and growth
0.01
f
responses. For this reason we describe 5 4 5 7 8 I0 12
the effects of light intensity on the
LIGHT INTENSITY (Kiux)
growth of Phaeodactylum tricornutum in batch
culture (Fig. 3). Comparable results
(with lower absolute growth rates) were Fig. 3. Phaeodactylum tricornutum. Relationship
observed in our turbidostat cultures. between light intensity and growth rate for
For subsequent p h o t o s y n t h e s i s experi- phytoplankton grown in batch culture
ments w i t h batch cultures the alga was
grown at 12 klux, 5 klux (both giving
m a x i m u m growth rate) and 0.7 klux (29.7%
of maximum growth rate). sons at a single time towards the end of
batch growth give a m i s l e a d i n g view.

Changing Photosynthetic Abilities During Growth


in Batch Culture Characteristics of Photosynthesis in Cultures
Grown at Various Light Intensities
Our earlier experiments (Morris and
Glover, 1974) suggested that the changes Fig. 6 shows light saturation curves for
in photosynthetic rates during batch photosynthesis by cultures of Phaeodactylum
growth were significant; that these tricornutum grown at 3 light intensities.
changes depended on the conditions under These experiments w e r e performed at
which the alga was grown; and that they those times which gave m a x i m u m rates of
should be taken into consideration in light-saturated photosynthesis in each
any attempts to u n d e r s t a n d so-called culture. W h e n expressed on a chlorophyll
"adaptation". Fig. 4 shows the changing basis, the Pmax was greater for cultures
rates of Pmax (light-saturated photosyn- grown at higher light intensities. When
thesis) by cultures grown at 12 and 0.7 expressed per cell, the values for Pmax
klux. The m a x i m u m rate'of photosynthesis were approximately the same. The most
occurred later (4 days) in the cultures interesting observation from these ex-
grown at the lower light intensity. This periments was the response of the vari-
means that, if the comparison is made ous cultures to low light intensities.
after 7 days, as in Steeman N i e l s e n and Cells grown at the lowest intensity were
J~rgensen's work (1968) with Chlorella able to utilize intensities b e l o w satura-
pyrenoidosa, cells grown at low light in- tion levels more efficiently than those
tensities show higher Pmax values than grown at saturating intensities. Only
those grown at higher light intensities. part of the explanation lay in the higher
This is comparable to the results of chlorophyll content, since w h e n ex-
Steeman N i e l s e n and J ~ r g e n s e n (1968) pressed on a chlorophyll basis, cells
presented in Fig. 2b of this paper. How- grown at the lowest light intensity uti-
ever, it can be seen that this conclu- lized low light levels more efficiently
sion is not valid if the earlier m a x i m u m than did the others. The difference be-
rate of photosynthesis in cultures at 12 tween cultures grown at 5 and 12 klux
klux is considered. Fig. 5 shows the was less marked. Although, therefore,
changing chlorophyll content during there appeared to be adaptation of the
batch growth (a) and the changing Pmax photosynthetic m e c h a n i s m to low light
(b) expressed on a chlorophyll basis. intensities, it is w o r t h emphasizing
Again, a comparison b e t w e e n the two cul- that the growth rate at these intensi-
tures is valid only w h e n early peaks of ties was less than at saturating levels
photosynthesis are considered; compari- (Fig. 3).
382 J. Beardall and I. Morris: Light Adaptation in Phytoplankton

6.5

5.5

~ 4.51 i_o
Z / I 0

.u
5.5 I I I I I I I

I00

ro
0
• 75
i

o o

(.0
_o
5O

N 25
E
a_

l I I I I I I
I 2 5 4 5 6 7
DAYS

Fig. 4. Phaeodactylum tricornutum. Changes in cell density (a) and rates of light-saturated photo-
synthesis (Pmax) per cell (b) for phytoplankton grown at 12 klux (closed circles) and 0.7 klux
(open circles) in batch culture, cpm: Counts per min

0 8OO

05 z-
T 70O
,.c
tJ
_J 0.4
w I 600
~o 0.5
o
~ ~ o 0_ ~ 5 0 0
c.)~
0.2 ~ 1 7 6 1 7 6 1 7 6 ~o
,~ '< 4 0 0
0.1 __s
~- :500
I I f i i I I

o
0
200
/
Fig. 5. Phaeodactylum tricornutum. Changes in ,- I00
o.
chlorophyll a content (a) and Pmax expressed on
I } I I ) i I
chlorophyll basis (b) for phytoplankton grown I 2 3 4 5 6 7
in batch culture at 12 klux (closed circles)
and 0.7 klux (open circles) DAYS
J. Beardall and I. Morris: Light Adaptation in Phytoplankton 383

o
• 500,
~

ii ill"
~ 40c

~ 50C

~ 20C

~ IOO ,>/;//
o
t 0
g- 0

120
b
Z~ ~.Z~.~" ' ' ' A ~ 0 0 ,......,~.,-~- 0 - " ~ " ~ ' ~ "
_o I00

% 80 / o,
/Y
o
"-c 60
s

v
~ 40

= 20

o
o
_ _ _ L t J _ _ _ • 1 l __L__
O_
0 2 4 6 8 I0 12 14 24
LIGHT INTENSITY (Klux)

Fig. 6. Phaeodactylum tricornutum. Light saturation curves for photosynthesis of phytoplankton


grown in batch culture at 12 klux (closed circles), 5 klux (open circles) or 0.7 klux (triangles).
Photosynthesis is expressed on per unit chlorophyll (a) and cell-number (b) basis

Experiments with Turbidostat Cultures levels of p h o t o s y n t h e t i c p i g m e n t s and


b u l k cell protein, Fig. 8 c o m p a r e s the
It is a p p a r e n t that c h a n g e s d u r i n g b a t c h rates of l i g h t - s a t u r a t e d p h o t o s y n t h e s i s
g r o w t h can o b s c u r e the r e s p o n s e of a l g a e w i t h the a c t i v i t i e s of RuDP c a r b o x y l a s e
to v a r i o u s l i g h t i n t e n s i t i e s . T h e s e prob- (expressing b o t h r a t e s on a c h l o r o p h y l l
lems can be o v e r c o m e by u s i n g c o n t i n u o u s and cell n u m b e r basis) and Fig. 9 com-
c u l t u r e s o p e r a t i n g as t u r b i d o s t a t s w i t h p a r e s Pmax, RuDP c a r b o x y l a s e a c t i v i t y
g r o w t h l i m i t e d by light i n t e n s i t y . In and m a l a t e d e h y d r o g e n a s e a c t i v i t y , ex-
our t u r b i d o s t a t e x p e r i m e n t s we have em- p r e s s i n g all on a p r o t e i n basis.
p h a s i z e d r a t e s of l i g h t - s a t u r a t e d p h o t o - The m a i n p o i n t s to e m p h a s i z e from
s y n t h e s i s r a t h e r than the r e s p o n s e of Figs. 7, 8 and 9 are the f o l l o w i n g :
the alga to low light levels. Our m a i n (a) G r o w t h at low light i n t e n s i t i e s
aim has b e e n to p u t the c h a n g i n g r a t e s p r o d u c e d h i g h e r c e l l u l a r c o n t e n t s of
of p h o t o s y n t h e s i s on an e n z y m a t i c b a s i s b o t h c h l o r o p h y l l a and c a r o t e n o i d s .
and to d e s c r i b e the w a y in w h i c h the T h e r e was l i t t l e c h a n g e in the p r o t e i n
v a r i o u s c o m p o n e n t s of the p h o t o s y n t h e t i c content. ( M e a s u r e m e n t s w i t h b a t c h cul-
m a c h i n e r y are i n f l u e n c e d by the light tures also s h o w e d a s l i g h t d e c r e a s e in
i n t e n s i t y used for growth. Figs. 7, 8 m e a n cell v o l u m e w i t h g r o w t h at the
and 9 s u m m a r i z e these data: Fig. 7 shows lower light intensity.)
384 J. Beardall and I. Morris: Light A d a p t a t i o n in P h y t o p l a n k t o n

(b) The c h a n g i n g Pmax v a l u e s w e r e


7.5 p a r a l l e l e d by a l t e r e d a c t i v i t i e s of RuDP
carboxylase. This was not part of gen-
eral changes in enzymes since the activ-
0.50 7.0
ity of m a l a t e d e h y d r o g e n a s e did not cor-
relate as c l o s e l y w i t h Pmax as did the
"T carboxylase.
65
(c) B e t w e e n 12 and I klux the m a j o r
change was a 39% i n c r e a s e in the cellu-
V m
6.0
lar c h l o r o p h y l l content. This was the
0.40
m cause of the o b s e r v e d r e d u c t i o n s (at the
o O
lower intensity) of Pmax and RuDP car-
-- E 5.5 b o x y l a s e w h e n e x p r e s s e d on a c h l o r o p h y l l
o
O
basis. At the two lower i n t e n s i t i e s (0.86
Z x, and 0.53 klux), however, there was a
~ 0
0 30 5.0 E
change in Pmax and RuDP c a r b o x y l a s e ac-
E ~
0 tivity per cell. That is, at these low
i n t e n s i t i e s the i n c r e a s e d c o n c e n t r a t i o n
i

\o !45 of c h l o r o p h y l l in the dell was accom-


p a n i e d by d e c r e a s e d levels of the en-
zymatic m a c h i n e r y r e s p o n s i b l e for the
0.20 4.0 dark r e a c t i o n s of p h o t o s y n t h e s i s .
I I I i

0 0.4 0.8 1.2 12


LIGHT INTENSITY ( K l u x )
Time Required for "Adaptive" Changes
Fig. 7. Phaeodactylum tricornutum. Changes in
cellular content of chlorophyll a (closed cir- We agree w i t h S t e e m a n N i e l s e n and J ~ r g e n -
cles), carotenoid (open circles) and protein sen (1968) in e m p h a s i z i n g the d i s t i n c -
(triangles) in cells grown at different light tion b e t w e e n g e n o t y p i c and p h e n o t y p i c
intensities in turbidostat cultures, m-sp-u: adaptation. W h e n o r g a n i s m s are grown
Millispecific plant pigment unit under d i f f e r e n t e n v i r o n m e n t a l c o n d i t i o n s

(0) PHOTOSYNTHESIS (b) RuDP CARBOXYLASE ACTIVITY


560 - 16r 350

480 - 141- 300


"7
/
I

-J

400- ~ 12 t 250
T-- %i

~'
.1:.
3zo- '~ lot
uJ
x I
/ / 200
O
~'
E

:/j/~
x_
u_
24o-
~
81-
,
150 x~

~J
g
~, i 6 o - ~ 6~ I00 ~
o
E
E I

80- 4 50

o /
o- zf I i
9 I I I ~ I I I I I

0.4 0.6 0.8 1.0 12.0 0.4 0,6 0.8 1,0 12.0

Fig. 8. Phaeodactylum tricornutum. Changes in (a) rate of light-saturated p h o t o s y n t h e s i s and (b)


RuDP carboxylase activity in cells grown at different light intensities in t u r b i d o s t a t cultures.
Data are expressed on chlorophyll basis (open circles) and per unit number of cells (closed cir-
cles)
J. Beardall and I. Morris: Light Adaptation in Phytoplankton 385

for several generations, strain selec- Discussion and Conclusions


t i o n -- as o p p o s e d to p h y s i o l o g i c a l
c h a n g e s t o t h e e n t i r e p o p u l a t i o n -- c a n
complicate interpretation of the data. The conclusions t o b e d r a w n f r o m t h e in-
In o u r e x p e r i m e n t s w e h a v e m a d e no c r i t - vestigation presented here fall into 4
i c a l a t t e m p t s to d i s t i n g u i s h between main categories.
t h e s e t w o t y p e s of r e s p o n s e s . However, F i r s t l y , a n a l y s i s of t h e p u b l i s h e d
the observations presented in Fig. 10 literature on light-intensity adaptation
suggest a physiological response. That suggests that too great an emphasis has
is, t h e t i m e t a k e n f o r c h a n g e s in c h l o r o - b e e n p l a c e d o n Ik, t h e l i g h t i n t e n s i t y
p h y l l c o n t e n t a n d in r a t e of l i g h t - s a t u - at w h i c h p h o t o s y n t h e s i s saturates and
rated photosynthesis per chlorophyll oc- t o o l i t t l e o n the c h a n g e s in Pmax. B e -
c u r r e d o v e r a t i m e p e r i o d e q u a l to (or c a u s e of this, it h a s b e e n i m p l i e d t h a t
l e s s then) a g e n e r a t i o n time. adaptation to s u b - o p t i m a l light inten-
sities involves some kind of enhanced

O~

~ 4.0

0
x I
v 3.5 !

I00 i,
o 3.0 &
90 7
II $ ~
E 2.5

80 iI/
a.0 II ~ e
._o I
7O
u) ~ 1.5
ILl
o
o

60 .c
a_ 1.0 J i l i I I I I I I I I
I-
,o
0.6
50
uJ
> b
t-
< 0.5
~, 40

--~ 0.4
50

_o
~ 0.3 ~.
20
o
o 0.2

I0 0
I I I I
0.4 0.6 0.8 1,0 120 0.1
LIGHT INTENSITY ( K l u x )

Fig. 9. Phaeodactylum tricornutum. Changing ~) I I I I I I I


0 2 4 6 8 I 12 14 16 18 20 22 24
rates of photosynthesis (closed circles), RuDP
TIME (HOURS)
carboxylase activities (open circles) and malate
dehydrogenase activity (triangles) in cells
Fig. 10. Phaeodactylum tricornutum. Changing
grown at different light intensities in turbido-
rates of light-saturated photosynthesis (a) and
stat cultures. All activities are expressed per
chlorophyll content (b) during 24 h following
unit amount of cell protein and are related to
transfer of cells grown at 0.7 klux to 12 klux
a value of "iOO" given for cultures grown at
(open circles) and following transfer from 12
12 klux
klux to 0.7 klux (closed circles)
386 J. Beardall and I. Morris : Light Adaptation in Phytoplankton

a b i l i t y to use the low light levels, c o n s i d e r e d only in terms of " g a i n i n g an


w h e r e a s it a p p e a r s that a r e d u c e d abil- ability". A r e g u l a t e d d i s a p p e a r a n c e of
ity to u t i l i z e s a t u r a t i n g light inten- specific enzymes under growth conditions
sities is the d o m i n a n t c h a r a c t e r i s t i c not u t i l i z i n g m a x i m u m a c t i v i t i e s is a
of algae g r o w n at low i n t e n s i t i e s . physiological adaptation which enables
Secondly, it is a r g u e d t h a t the data the c e l l u l a r m e t a b o l i s m of an o r g a n i s m
of S t e e m a n N i e l s e n and J J r g e n s e n (1968) to r e s p o n d to c h a n g e s in the e n v i r o n m e n t .
for Chlorella pyrenoidosa m i g h t be d i f f i - Our p r e v i o u s p a p e r (Morris and Glover,
cult to i n t e r p r e t b e c a u s e of c h a n g i n g 1974) on t e m p e r a t u r e a d a p t a t i o n and the
photosynthetic abilities during growth p r e s e n t data argue for this m o d i f i e d
in b a t c h culture. T h e s e w o r k e r s o b s e r v e d v i e w of a d a p t a t i o n in p h y t o p l a n k t o n .
i n c r e a s e d c h l o r o p h y l l levels in cells H i t h e r t o , the a p p r o a c h has g e n e r a l l y been
grown at low light i n t e n s i t i e s and re- c o n c e r n e d w i t h the way in w h i c h o r g a n i s m s
p o r t e d e n h a n c e d rates of p h o t o s y n t h e s i s a d a p t to s u b - o p t i m a l c o n d i t i o n s . A shift
at all light i n t e n s i t i e s per cell (but of e m p h a s i s such as is s u g g e s t e d here
not per chlorophyll) by such a l g a e from m a k e s one ask q u e s t i o n s a b o u t the bio-
low i n t e n s i t i e s . We o b t a i n e d c o m p a r a b l e c h e m i c a l c h a n g e s r e q u i r e d of p h y t o p l a n k -
r e s u l t s w h e n the m e a s u r e m e n t s w e r e m a d e ton w h e n they m o v e into c o n d i t i o n s al-
at a single time t o w a r d s the end of the lowing m a x i m u m growth; s a t u r a t i n g light
g r o w t h curve. However, w h e n the c h a n g i n g i n t e n s i t i e s , o p t i m u m t e m p e r a t u r e s , and
photosynthetic abilities during batch possibly saturating nutrient concentra-
g r o w t h w e r e taken into account, a dif- tions. It is i n t e r e s t i n g that D u g d a l e
f e r e n t r e s u l t was obtained. (personal c o m m u n i c a t i o n ) is a s k i n g com-
Thirdly, w h e n p h o t o s y n t h e s i s m e a s u r e - p a r a b l e q u e s t i o n s c o n c e r n i n g the "shift-
m e n t s are m a d e at the time of m a x i m u m up" p r o c e s s e s w h i c h m a y occur in p h y t o -
photosynthetic ability during batch p l a n k t o n in r e s p o n s e to h i g h e r n u t r i e n t
growth, a d a p t a t i o n to light i n t e n s i t y levels and w h i c h m a y be b a s i c to an un-
t h r o u g h e n h a n c e d u t i l i z a t i o n of low d e r s t a n d i n g of the i n i t i a t i o n of p h y t o -
light levels can be observed. This is p l a n k t o n blooms.
m o s t m a r k e d w h e n p h o t o s y n t h e s i s is ex-
p r e s s e d per cell number, but is also
Acknowledgements. This work was supported in
a p p a r e n t w h e n e x p r e s s e d on a c h l o r o p h y l l
part by NSF Grant No. DFS 75-15104 and the Maine
basis. In our view, this is the first
Department of Marine Resources Contract No. 4-74.
p u b l i s h e d e v i d e n c e s u g g e s t i n g that g r o w t h
One of us (J.B.) acknowledges receipt of a re-
of an alga at r e d u c e d light levels en-
search studentship of the Science Research Coun-
h a n c e s its a b i l i t y to p h o t o s y n t h e s i z e at
cil.
s u b - o p t i m a l i n t e n s i t i e s and r e d u c e s its
a b i l i t y to u t i l i z e s a t u r a t i n g levels. It
is w o r t h e m p h a s i z i n g , however, that this Literature Cited
e n h a n c e d p h o t o s y n t h e t i c a b i l i t y does not
r e s u l t in i n c r e a s e d g r o w t h rates; g r o w t h Aruga, Y. and S. Ichimura: Characteristics of
at 0.7 k l u x b e i n g 29.7% of the m a x i m u m photosynthesis of phytoplankton and primary
rate. production in the Kuroshio. Bull. Misaki
F o u r t h l y , it has b e e n p o s s i b l e to mar. biol. Inst. Kyoto Univ. 12, 3-20 (1968)
relate changing photosynthetic abilities Beardall, J. and I. Morris: The effects of en-
to c h a n g i n g a c t i v i t i e s of e n z y m e s r e s p o n - vironmental factors on the patterns of photo-
sible for the dark r e a c t i o n s of p h o t o - synthesis in the marine diatom Phaeodactylum
synthesis. Over a w i d e i n t e n s i t y r a n g e tricornutum. II. The effect of oxygen. J.
(12 to I klux) the RuDP c a r b o x y l a s e lev- Phycol. 11, 430-434 (1975)
el per cell r e m a i n e d m o r e or less con- Bjorkmann, O.: Carboxydismutase activity in
stant. However, over this same r a n g e the shade-adapted and sun-adapted species of
ratio of c a r b o x y l a s e : c h l o r o p h y l l de- higher plants. Physiologia PI. 21, i-iO
c r e a s e d b e c a u s e of an i n c r e a s e in c h l o r o - (1968)
p h y l l content. At lower i n t e n s i t i e s , Glover, H.E., J. Beardall and I. Morris: The
there was a r e d u c e d a c t i v i t y of RuDP car- effects of environmental factors on the pat-
b o x y l a s e per cell and per unit a m o u n t terns of photosynthesis in the marine diatom
of cell protein. C o m p a r a b l e r e d u c t i o n s Phaeodactylum tricornutum. I. The effect of
in RuDP c a r b o x y l a s e levels have b e e n ob- nitrogen deficiency and light intensity J.
served in s h a d e - a d a p t e d s p e c i e s of h i g h e r Phycol 11, 424-429 (1975)
p l a n t s (Bjorkmann, 1968). Guillard, R.R.L. and J.H. Ryther: Studies of
This d e c r e a s e in the level of the en- marine planktonic diatoms. I. Cyclotella nana
zymatic m a c h i n e r y r e q u i r e d for m a x i m u m Hustedt and Detonula confervacea (Cleve) Gran.
u t i l i z a t i o n of s a t u r a t i n g light inten- Can. J. Microbiol. 8, 229-239 (1962)
sities can be seen as an a d a p t i v e re- Ichimura, S., Y. Saijo and Y. Aruga: Photosyn-
sponse to g r o w t h at l i m i t i n g light lev- thetic characteristics of marine phytoplank-
els, i.e., " a d a p t a t i o n " s h o u l d not b e ton and their ecological meaning in the chlo-
J. Beardall and I. Morris: Light Adaptation in Phytoplankton $87

rophyll method. Bot. Mag., Tokyo 75, 212-220 - and E.G. J~rgensen: The adaptation of plank-
(1962) ton algae. I. General part. Physiologia PI.
Lorenzeni C.J.: Determination of chlorophyll 21, 401-413 (1968)
and phaeopigments: speetrophotometric equa- Talling, J.F.: Photosynthetic characteristics
tions. Limnol. Oeeanogr. 12, 343-346 (1967) of some freshwater plankton diatoms in rela-
Morris, I. and H.E. Glover: Questions on the tion to underwater radiation. New Phytol. 56,
mechanism of temperature adaptation in marine 1-132 (1957)
phytoplankton Mar. Biol. 24, 147-156 (1974) Yentsch, C.S. and R.W. Lee: A study of photosyn-
- and P.J. Syrett: The effect of nitrogen star- thetic light reactions and a new interpreta-
vation on the activity of nitrate reductase tion of sun and shade phytoplankton. J. mar.
and other enzymes in Chlorella. J. gen. Res. 24, 319-337 (1966)
Microbiol. 38, 21-28 (1965)
Parsons, T.R. and J.D.H. Strickland: Discussion Dr. I. Morris
of spectrophotometric determination of marine Bigelow Laboratory for Ocean
plant pigments with revised equations for Sciences
ascertaining chlorophylls and carotenoids. Division of Northeastern
J. mar. Res. 21, 155-163 (1963) Research Foundation
Steeman Nielsen, E. and V.K. Hansen: Light adap- McKown Point
tation in marine phytoplankton populations West Boothbay Harbor, Maine
and its interrelation with temperature. 04575
Physiologia PI. 12, 353-370 (1959) USA

Date of final manuscript acceptance: May 28, 1976. Communicated by M.R. Tripp, Newark