Arch. Mikrobiol.

78, 25--41 (1971)

9 by Springer-Verlag 1971

Growth and Photosynthesis in an Extreme Thermophile,

Synechococcus Lividus (Cyanophyta)
Jo~ C. M ~ K S a n d Rm~rARD W . CAST~,~CTrOLZ
Department of Biology, University of Oregon, Eugene, Oregon 97 403, USA

Received February 18, 1971

Summary. A high temperature strain of the blue-green alga, Synechococcu8

lividus has been cultured and cloned in defined medium.
1. S. lividus (Culture OH-68-s, Clone H-X]) is an obligate thermophile with a
temperature range of growth from 54 to 72~C. The optimal conditions for growth
were at 63 to 67~ and a light intensity greater than 700 ft-c, resulting in a
reproducible minimum generation time of 11 hours. Growth was depressed in the
supra-optimal range from 68 to 72~ The temperature characteristic or coeffi-
cient (#) of growth was calculated as 13,750 cal mole-1. This value would not
distinguish this organism from mesophilic and psychrophilic yeasts and bacteria.
2. Clone H-X] photosynthesized from as low as 33 to 75~ during short
exposure times (20 rain) without prior acclimation to the incubation temperatures.
Longer exposures to the higher temperatures indicated that the "stable" upper
limit for photosynthesis was 73~C when cells were grown above 60~C, but was only
70~ for cultures grown at 55 and 57~C.
3. Abrupt shifts of exponential cultures between optimal(65~C) and near minimal
(55~C) growth temperatures showed that lag periods occurred before normal growth
commenced at the new temperatures. However, these lag periods on downward
temperature shifts followed only after a period of residual growth. Similar shifts
from optimal to subminimal (45~C) temperatures indicated that growth continued
for a period of time before entering an ex~nded stationary phase prior to senescence.
Both of the latter types of experiments may indicate that products synthesized at
65~ are consumed by residual growth after the shifts to lower temperatures, but
that these are replaced after a long delay (acclimation) at 55~C and not at all at 45~C.
4. Photoincorporation of ltC-NaHCOs was highly sensitive to subminimal tem-
peratures after the first hour of exposure. The data suggest that the photosynthetic
system could be involved in determining both the upper and lower limits of growth
in this organism.

A n u m b e r of thermophilie blue-green algae h a v e been isolated a n d

c u l t u r e d u n d e r ]aboratory conditions (Castenholz, 1969, 1970). Experi-
m e n t a l use has b e e n m a d e of thermophilie Oscillatoria (Cas~enholz, 1968,
1971), Mastigocladus (Holton, 1962), a n d several strains of Synechococcus
(Berns a n d Scott, 1966; D y e r a n d Gafford, 1961; E d w a r d s et al., 1968;
P e a r y a n d Castenholz, 1964; S h e r i d a n a n d Castenholz, 1968). Dyer a n d
Gafford (1961) first reported the isolation of a s t r a i n of Synechococcus
lividus t h a t grew u p to 55 ~C. However, field investigations b y Castenholz
26 J.C. Meeks and R. W. Castenlaolz:

(1969), B r o c k (1967a), a n d K e m p n c r (1963) h a v e i n d i c a t e d t h a t t h e u p p e r

t e m p e r a t u r e l i m i t of g r o w t h of p h o t o s y n t h e t i c blue-green algae lies
b e t w e e n 73 a n d 75 ~C. P e a r y a n d Castenholz (1964) r e p o r t e d t h e isolation
a n d g r o w t h o f four t e m p e r a t u r e s t r a i n s of S. lividus in enriched h o t spring
w a t e r . One s t r a i n (Strain I V) p r o b a b l y r e p r e s e n t s t h e h i g h e s t t e m p e r a t u r e
p h o t o s y n t h e t i c o r g a n i s m in h o t springs a n y w h e r e in t h e world.
The temperature characteristic or coefficient of growth (#) has been used in
attempts to define psychrophilic or mesophilic tendencies of microorganisms (see:
Baig and Hopton, 1969; Ingraham, 1958). I t is defined by the Arrhenius equation:
log of the rate = /~/2.303 RT + C, where _R is the gas constant, T the absolute
temperature, and C a constant. Thus, # is equivalent to the activation energy of a
chemical equation. When the log of the growth rate of an organism is plotted against
the reciprocal of the absolute temperature in the manner of the Arrhenius equation,
there is often a range of" temperature over which the growth rate increases linearly
with temperature. Generally, # is calculated from the slope of the straight line of best
fit of the experimental points in the middle portion of such a plot where the slope is
essentially straight (Shaw, 1967). The temperature range where # is relatively
constant includes most of the suboptimal temperature range of most micro-
organisms. Throughout the optimal temperature range of growth # values approach
zero, near the upper temperature limit they become negative, and near the minimal
growth temperature # values increase rapidly.
Ng et at. (1962), Shaw (1967), and Shaw and Ingraham (1967) have noted the
importance of the types of # values when examining the responses of heterotrophic
microorganisms to abrupt shifts in environmental temperatures during exponential
growth. In the mesophilic bacterium Escherichia coli M L 30 (Ng et al., 1962; Shaw
and Ingraham, 1967) and mesophilic and psychrophilic yeasts (Shaw, 1967), sudden
temperature shifts within the range of constant/l resulted in an immediate change
of the specific growth rate to that of the new temperature. Shifts between the range
of rapidly increasing # and the range of constant # resulted in lag periods and inter-
mediate growth kinetics.
W e h a v e r e c e n t l y isolated in defined m e d i u m a t 70~ a high t e m p e r -
a t u r e s~rain o f Synechococcu8 lividus, similar t o S t r a i n I V of" P e a r y a n d
Castenholz (1964). This p a p e r r e p o r t s t h e basic g r o w t h c h a r a c t e r i s t i c s of
t h e new isolate, its specific t e m p e r a t u r e c h a r a c t e r i s t i c (#) w i t h r e s p e c t t o
b o t h g r o w t h a n d p h o t o s y n t h e s i s , a n d t h e t e m p e r a t u r e range for b o t h of
these processes. W e h a v e also shown t h a t t h e responses o f this o r g a n i s m
t o a b r u p t t e m p e r a t u r e shifts differ c o n s i d e r a b l y from those of mesophilie
a n d p s y c h r o p h i l i c y e a s t s a n d bacteria, a n d o f t h e r m o p h i h c bacteria.

Materials and Methods

I s o l a t i o n a n d Cloning
Unicellular blue-green algae fitting the description of Synechococcu8 lividus Cope-
land (1936) were collected in 1968 from substrate covered by water of over 70~ C
in one of the hot spring drainways at Hunter's Hot Springs, Lakeview, Oregon
(pH 8.2 to 8.5). These are blunt-ended unicellular rods with a width of i.2 to 1.8 ~m
and a more variable length, seldom less that 5 to 7 ~zm. The algae were enriched at
70~ and 600 ft-e in defined medium D (pI-I adjusted to 8.5 before autoclaving)