ANNUAL REVIEWS

Further

Quick links to online content

TEMPERATURE EFFECTS ON MICROORGANISMSl.2.3

By
JUDITH FARRELL AND ANTHONY ROSE

Department of Microbiology, University of Newcastle upon Tyne, England

CONTENTS

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

METHODOLOGY............................................... . .... THERMOPHILES . . .. ............................... ............ .." PHYSIOLOGICAL BASIS OF THERMOPHILISM. ....................... ...... PSYCHROPHILES................................................... DEFINITION OF PSYCHROPIIILISM.. . . . . ................................ ECOLOGY AND ISOLATION . ......................... ........... .... . " DISTRIBUTION OF THE PSYCHROPHILIC HABIT AMONG MICROORGANISMS. ... MICROBIAL METABOLISM AT SUBOPTIMUM TEMPERATURES.... PHYSIOLOGICAL ACTIVITIES. . . . ....................................... Pigment production .... .... " .................................... " Flagella production .. .... . ....... ......... ... ...... ...... . " Polysaccharide synthesis ....... .................... . .... ....... "
. . . . . . . . . . .

102 103 104 106 106

107 108 110 110 110 111 111 Fatty acid synthesis. . ............................... ........... . ... 111 Solute uptake ....................................... .. .. ........ " 112 SYNTHESIS AND ACTIVITY OF ENZYMES. .......................... . ..... 114 Total protein synthesis. ... .. ... .... .................. ............ " 114 Synthesis of individual enzymes. ........ ................ ............. 114 TEMPERATURE SENSITIVE MUTANTS ............ . . .......... .. .. 115
.

After accepting this reviewing assignment, we were genuinely surprised to discover what little coverage had been given to the effects of temperature on microorganisms in previous editions of Annual Review

of Microbiology. No

article dealing specifically with this subject has appeared before in the series, despite the fact that temperature is one of the most important environmen tal factors affecting the growth and development of microorganisms, and that the evolution of microbiology as a scientific discipline depended in no small way on the discovery of suitable methods of heat sterilization. These comments should not be construed as criticism of past editorial committees. Rather, they reflect the lack of appreciation by microbiologists-at least until quite recently-of the full implications of the effects of temperature on microorganisms, particularly the manner in which temperature influences the activities of microorganisms in natural environments and, more recently, the ways in which temperature-sensitive mutants can be used in studies on metabolic regulation. We felt that this review would be of greatest value to microbiologists if we confined our attention to a small number of topics rather than attempt to
1

The survey of the literature pertaining to this review was concluded in December

I Experimental work from this laboratory was supported by grants from the Science Research Council, The Nuffield Foundation, and The Royal Society.

1966. I All temperatures quoted in this review are in degrees Centigrade.

101

102

FARRELL &

ROSE

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

cover the field as a whole with its voluminous and scattered literature. Several topics which we have not been able to cover in the present article have been reviewed recently (1-3). The effects of superoptimum as well as near-zero and subzero temperatures on the viability of microorganisms have continued to receive a great deal of attention, both from workers interested in the physiological bases of these stresses (3, 4-19) and from microbiologists more concerned with the applied aspects of the subject [for a review see (2)]. Several papers have appeared recently dealing with the effects of temp erature on differentiation (20-24), rate of mutation, and genome activity in microorganisms (4, 25-30). But the most active area of research continues to be the growth pf spoilage microorganisms on heat-sterilized and on chilled and frozen materials, particularly foodstuffs. Fortunately, the extensive literature on these subjects has been reviewed quite regularly in recent years (31-36). METHODOLOGY The reader might be excused for assuming that the methodology involved in studying the temperature responses of microorganisms is sufficiently straightforward and uncomplicated that it could hardly justify special com ment in a review of this nature. Nevertheless, it is clear that this is not so, and that the widespread lack of detailed information on the effects of temp erature on the growth and activities of microorganisms can be traced in many instances to the fact that few laboratories are able to grow micro organisms under carefully controlled conditions at a large number of different temperatures. If temperature-response curves are to be described accu rately, it is absolutely essential that cultures or reaction mixtures be in cubated at temperatures that vary by not more than O.2. It is generally agreed that anhydric incubators are unsatisfactory for this purpose, and re sort has to be made to constant-temperature water baths, many varieties of which are available commercially. Additional problems arise when cultures need to be aerated or stirred at constant temperature. Ingraham and his colleagues (37, 38) have solved this problem by using glass test tubes, measuring about 300 mm by 30 mm in diameter, and immersing these in water baths at constant temperature; the tube cultures are aerated by bubbling in sterile air. This method has the great advantage of being very economical on water-bath space. Extensive use has been made in our labora tory of 2-1 and 5-1 flat-bottom flasks which are jacketed in Perspex baths through which is circulated water at constant temperature. The flasks con tain a 4 em or 6 em follower magnet, and the contents are stirred magneti cally (39). The problem of procuring a large number of different temperatures has been solved in some laboratories by using temperature-gradient incubators. A number of different designs have been described, but all temperature gradient incubators consist basically of bars of heavy gauge metal at one end of which heat is applied. This heat is dissipated along the length of the bars

TEMPERATURE ON MICROORGANISMS

103

so that a temperature gradient is set up, the magnitude of which can be con trolled by regulating the amount of heat applied, by varying the efficiency of the insulating material, and by controlling the ambient temperature . The culture tubes are usually placed in holes in the metal bar. An interesting development in the design of temperature-gradient incubators has been de scribed by Nakae

(40) who has constructed an apparatus in which a tempera

ture gradient is maintained by heat conduction in a liquid such as water. Tubes containing cultures on solid media are inserted vertically into the temperature-gradient bath, and visible growth of the organism appears in definite sections of the agar column, corresponding to the temperature range for growth . Most temperatue-gradient incubators can be used only with organisms growing on solid media, although modifications have been made to some instruments so that they can be used with liquid cultures of temperature on growth of bacteria

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

(41) .

Temperature-gradient incubators have been used for studying the effects

(41-45), yeasts (46, 47), and algae (48). 0.2, and these instruments

With many temperature-gradient incubators, it is possible to incubate cul tures at temperatures differing by as little as have proved useful in studies on the effect of superoptimum temperatures on growth of microorganisms. For example, Elliott & Heiniger the maximum temperatures for growth of tween growth of a strain of

(44) showed that 34 strains of Salmonella were be

43.2 and 46.2. In another study (46), the maximum temperature for Saccharomyces cerevisiae was shown to be 40.90.4 when the yeast was grown aerobically, and 40.3 0.6 when grown anaerobi callYi the corresponding minimum temperatures for growth were 9.60.9 and 9.7 0.3. Accurate data of this type are urgently required in studies on
the physiological bases of the maximum and minimum temperatures for

growth of microorganisms. THERMOPHILES Microorganisms which are capable of growing at temperatures above

50

are described as thermophilic . Among the earliest thermophiles to be recog nized were algae that are found in geysers and hot springs in Yellowstone N a tional Park, Wyoming, and from similar habitats in New Zealand, Iceland, and Japan. These algae are almost all members of the Cyanophyta (blue green algae), and include species of

Oscillatoria and Synechococcus (49) . The

principal reason for interest in these thermophilic algae has been their re portedly very high maximum temperatures for growth. Some of the values reported in the literature for the maximum growth temperatures of these organisms are undoubtedly exaggerated . It has been claimed, for example, that blue-green algae from geysers in Yellowstone National Park have a maximum temperature for growth as high as

89, but

a recent

reinvestigation

of these algae has shown that the maximum temperature is nearer boils at around 93 .

It is worth noting that, at the elevation of Yellowstone National Park, water

75 (50) .

While the thermophilic algae have attracted attention because of their

104

FARRELL & ROSE

ability to grow at very high temperatures, it is to the thermophilic bacteria that many microbiologists have, perforce, turned because of their commer cial importance, principally as spoilage organisms in heat-treated foods and food materials. Valuable reviews on thermophilic bacteria have come from Gaughran (51) and Allen (52). In recent years, studies with these organisms have been f<,>cused mainly on the physiological basis of thermophilism (which is discussed below) and on the effects of high temperatures on the viability of thermophilic bacilli and their spores (53-56). McDonald & Matney (57) have examined the genetic basis of the ability to grow at 55 in strains of Bacillus subtilis. When strains which lacked the ability to grow above 50 were grown in the presence of DNA extracted from a strain capable of grow ing at 55, the ability to grow at the higher temperature was transformed at a frequency of 10-4 An incubation period of 3 to 4 hr at 37 was necessary for phenotypic expression of the ability to grow at 55. The character conferring ability to grow at 55 appeared to be closely linked to the single-step, high level streptomycin-resistance locus. Waksman & Corke (58) have discussed the taxonomy of the thermophilic actinomycetes. The genus Thermoactinomyces was created by Tsiklinsky in 1899 to accommodate the monosporous actinomycetes that are capable of growing at high temperatures. In addition, there exist thermophilic strains of Streptomyces (59), and some isolates of Nocardia sebivorans can withstand a temperature of 90 for 10 min (60). On the whole, little appears to be known about the thermophilic actinomycetes, and these organisms merit more de tailed consideration from microbiologists. For many years, thermophilic fungi too were blatantly neglected. Al though the first description of a thermophilic fungus was made in 1886 by Lindt (61), as late as 1963 the number of published descriptions of thermo philic fungi was still only six. But, as a result of the publication of a mono graph by Cooney & Emerson (62), a delightfully produced volume the like of which is a rarity in these days of rapid publication, our knowledge of ther mophilic fungi has been considerably embellished. Cooney & Emerson de scribed thermophilic fungi isolated from composting plant materials. They confined the epithet "thermophilic" to fungi that have a maximum tempera ture for growth of 50 or above, and a minimum temperature above 20. Thirteen strains of thermophilic fungi were examined, including four pre viously undescribed species and three new varieties of existing species. They included strains of Chaetomium, Humicola, Malbranchea, Myriococcus, Mucor, Sporotrichum, Talaromyces, Thermoascus, and Torula.
PHYSIOLOGICAL BASIS OF THERMOPHILISM

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

Undoubtedly, the most intriguing aspect of the microbiology of thermo philes is the physiological basis of their ability to grow at high temperatures. Most of the investigations in this area have been carried out using thermo philic bacilli. Earlier work on this topic was reviewed by Gaughran (51) and Allen (52), and later critically analyzed and extended by Koffler (63).

TEMPERATURE ON MICROORGANISMS

lOS

It is now generally agreed that the capacity of thermophilic bacteria to grow at high temperatures is due to their being able to synthesize cell constit uents that have a heat stability that is much greater than that of the cor responding cell components synthesized by other microorganisms. Several enzymes from thermophilic bacteria have been reported to possess excep tional heat stability; they include malate dehydrogenase (64), inorganic pyrophosphatase (65, 66), ATPase (67), and aldolase (68); Campbell and his colleagues have also shown that the hydrogenase (69) and the sulphate adenyJyJ transferase (70) from Desulfotomaculum nigrificans [once called Clostridium nigrificans (71)] are more heat-stable than the corresponding enzymes from the mesophile, Desulfovibrio desulfuricans. Almost all of these experiments were done with unfractionated cell ex tracts, and they do not show whether the heat stability is an intrinsic prop erty of the enzymes, or whether it is brought about through the action of protective factors in the cell or cell extract. To obtain information on these possibilities, Manning & Campbell (72) purified and crystallized the heat stable extracellular a-amylase of Bacillus stearothermophilus. The crystalline enzyme was electrophoretically homogeneous and extremely resistant to heat inactivation. The heat stability of this enzyme appears, therefore, to be an intrinsic property of the protein molecule. Flagella from thermophilic strains of Bacillus show a much greater heat stability than those from a mesophilic strain of E. coli (13), and the additional heat stability of these flagella also appears to reside in the protein (flagellin) monomers (73). Flagella from a strain of B. stearothermophilus were disrupted at pH 2.3 and the monomers thus obtained reassembled at high temperatures (58) to give filaments that resembled native flagella, thereby providing addi tional evidence for the heat stability of the component proteins. Other investigators have examined the heat stability of the components of the protein-synthesizing systems in thermophilic bacilli. The melting tem perature (Tm value) of the DNA from a thermophilic bacillus was comparable to that of DNAs from mesophilic microorganisms (74), which suggests that there is no relationship between the thermal stability of the DNA and the ability of the thermophile to grow at higher temperatures than mesophiles. Increased heat stability has, however, been demonstrated with the ribosomal RNA from a thermophilic bacillus as compared with that from the mesophile E. coli (75,76). Moreover, the heat stability of the mesophile RNA was greater when the nucleic acid was in the ribosome rather than in the isolated state (76). Analyses of the ribosomes from the thermophile and the mesophile re vealed only slight differences in overall composition and would probably not account for the differences in the heat stabilities of the intact organelles (77). The greater heat stability of the thermophile ribosomes may result from a different packing of the protein in the organelle. Alternatively, it may be a reflection of differences in the composition and structure of the ribosomal protein although, since the proteins from the two types of ribosome had very similar amino acid compositions, any such differences must be rather subtle.

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

1 06

FARRELL & ROSE PSYCHROPHILES

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

The existence of microorganisms that are capable of growing well at near zero temperatures, (0 to 5), i .e., psychrophiles, has been recognized for al most a century. Nevertheless, until just over a decade ago, psychrophiles were a relatively neglected group of microorganisms. There have been two main reasons for the recent upsurge of interest in psychrophiles, namely, the increasing realization of the economic importance of these organisms as food spoilage organisms,and the desire to know more about the activities of micro organisms in natural environments such as soil and the oceans whose temp eratures are usually low. The earlier literature on psychrophilic microorga nisms has been summarized in a number of reviews (1-3, 78, 79), and the present review, insofar as it pertains to bacteria, will be confined largely to work that has been published since about 1955, when the excellent literature survey made by Ingraham & Stokes (7S) was completed.
DEFINITION OF PSYCHROPHILISM

Although the term "psychrophile" has been in use for over half a century, microbiologists have until recently been unable to agree on a precise defini tion of the term. Psychrophiles are a subdivision of the microbial kingdom,
along with mesophiles and thermophiles. Since representatives of the last two

groups of microorganisms are defined on the basis of their optimum tem peratures for growth,it seemed appropriate to the earlier microbiologists that psychrophiles be similarly characterized. Indeed, several standard microbio logical texts define psychrophiles as microorganisms with an optimum tem perature for growth below about 15 (SO,81). One of the main proposals made by Ingraham & Stokes (7S) was that psychrophiles should be defined, not on the basis of their optimum temperatures, but on their minimum tempera tures for growth. They suggested that psychrophiles be defined as micro organisms which grow well at 0 within two weeks. As a rider they suggested that good growth be considered as the formation on solid media of colonies visible to the naked eye. Stokes (82) later suggested that the period of incu bation might be shortened to one week at 0, and this is the definition of psychrophilism that will be adopted for the purposes of this review. Ingra ham & Stokes (78) and Stokes (82) deferred to the widespread use of optimum growth temperatures in defining psychrophilic microorganisms by suggest ing that these temperatures should continue to be used for subdividing the group into "obligate" psychrophiles, which have an optimum temperature for growth below 20 (i.e., the textbook definition) and "facultative" psychrophiles which have optimum temperatures above 20. This subdivi sion is useful and is now widely accepted. The vast majority of psychrophiles that have so far been described are facultative psychrophiles, but this may not reflect the true extent of their distribution in nature (see p. 107). A whole range of alternative terms for describing psychrophilic microorganisms have been proposed over the years (2,78, 79,83),but none of these has gained wide acceptance.

TEMPERATURE ON MICROORGANISMS
ECOLOGY AND ISOLATION

107

PsychrophiJes are ubiquitous and are not confined to environments which are normally at near-zero temperatures. It is quite evident, too, that, in con trast to the view held for many years, psychrophiles constitute a large and widespread group of microorganisms, probably larger and more widespread than the mesophiles. Although psychrophilic bacteria, yeasts, moulds, and algae have been isolated from a wide range of environments, information on the quantitative distribution of these organisms in different environments has only recently been reported. In a timely study, Stokes & Redmond (84) examined samples of soil, water and mud, and various foods, for their con tents of psychrophilic bacteria and fungi. Psychrophiles accounted for 0.5 to 86 per cent of the bacterial populations of the soils examined, the proportion being smallest in cultivated soils and greatest in uncultivated soils. Of the fungi isolated from uncultivated soils, 25 per cent were psychrophilic. In stream and river water, psychrophilic bacteria accounted for 16 to 47 per cent of the total bacterial population; in lake water, 41 to 76 per cent; and in lake mud, 11 to 33 per cent. Between 33 per cent and 93 per cent of the bac teria isolated from meats were psychrophilic. The distribution of psychrophilic microorganisms has been studied most intensively with materials from environments that are continually subjected to low temperatures. Freezing and cooling are used very extensively to pre serve perishable materials, particularly foodstuffs. At these temperatures (-10 to S), the growth of pathogenic and potentially pathogenic micro organisms is prevented (2, 3, 31-34, 36). Nevertheless, under these condi tions, psychrophilic microorganisms can grow freely. Not only are near-zero temperatures favourable for growth of psychrophiles, but these organisms are free from the competition of mesophiles. In addition, the food materials on which they grow usually provide a plentiful supply of nutrients. Growth of psychrophiles on chilled and frozen foods renders the food less acceptable for consumption, and can affect its organoleptic qualities, often as a result of the production of proteolytic enzymes by the very widespread psychrophilic pseudomonads (85). Fortunately, the psychrophiles that are found on chilled and frozen foods have not been found to produce toxins or toxic substances. Serious concern has, however, been shown following the report that spores of Clostridium botulinum type E can germinate, and the bacteria grow vegeta tively, around S (86-88). A keen interest has also been shown in the microbiology of the polar re gions. On the whole, the Antarctic continent and the areas surrounding it, have been accorded the greater interest, partly because they are the coldest on earth and therefore might be expected to support microorganisms adapted to extreme cold, but also because of the introduction of carefully planned and controlled programmes of scientific research in this area following the mora torium on territorial claims in 1959. The presence of bacteria, yeasts, and moulds in the polar regions has been known for some years (89-96). The

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

108

FARRELL & ROSE

majority of earlier reports on the microbial flora of the polar regions was con cerned with the isolation of different types of organisms, and little attention was given to the ways in which the organisms had become adapted to the rigours of the Antarctic environment. Some of the bacteria isolated from Antarctic sources were almost certainly introduced as a result of human activity

(97). But it was not until 1960 that Straka & Stokes (98) reported (99,100) reported the

that many of the bacteria isolated from samples of soil and glacier ice from the Antarctic were psychrophilic. This and later papers isolation of obligately psychrophilic yeasts from the Antarctic. The failure of workers to isolate obligately psychrophilic bacteria from this region was

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

emphasized, and led to the suggestion that such bacteria might be very un common

(100). Recent work from our laboratory suggests that this is not so,

and that the failure of workers to isolate obligately psychrophilic bacteria from Antarctic (and, indeed, other) materials may have been due to defi ciencies in technique. Stanley & Rose

(101) examined the bacterial and yeast

flora of five lakes on Deception Island in the Antarctic. During the sampling and isolation, which was done with the membrane-filter technique, special measures were taken to ensure that the microorganisms never experienced a temperature above

10. All of the bacteria isolated were psychrophilic, and

the majority obligately so. It would seem that many workers have paid in sufficient attention to the temperature during isolation, and so might have lost any obligatel y psychrophi lic bacteria in the samples being analyzed. Approximately three-quarters of the world's surface is covered by oceanic water, the bulk of which rarely reaches a temperature of 5 This is probably
50,

(102). One would

therefore expect the majority of marine microorganisms to be psychrophilic. but there is an astonishing dearth of data on the subject. This situation is largely the result of the lack of suitable incubation equip ment aboard ship. In addition, the effect of hydrostatic pressure on the tem perature relationships of marine microorganisms, which could be of para mount importance in the marine environment, has yet to be evaluated. Marine bacteria

(102-105) and yeasts (106) have been described which grow

well at low temperatures, although many of these reports do not indicate whether the organisms are able to grow well at

0. In addition, Bunt & Wood (107) have reported the isolation of diatoms from antarctic sea ice. But, in

reviewing the literature on the temperature relationships of marine micro organisms, one cannot fail to be struck by the paucity of published data. Clearly, this is an area that is in urgent need of study. DISTRIBUTION OF THE PSYCHROPHILIC HABIT AMONG MICROORGANISMS Some genera of microorganisms appear to be far better endowed than others with strains that are able to grow well at 0. Surveys of psychrophilic bacteria in different environments

(1-3, 35, 79, 101) show the predominance,

qualitatively and quantitatively, of Gram-negative, rod-shaped bacteria. The majority of these are pseudomonads; Pseudomonas geniculata is one of the most frequently isolated

(108), together with P. putrefaciens, P. jragi,

TEMPERATURE ON MICROORGANISMS

109

and P. fluorescens (79, 109). Also fairly common are psychrophilic strains of Flavobacterium (110-114), Alcaligenes (113, 115), Achromobacter (116), and Arthrobacter (117, 118). Less frequently, but nevertheless regularly, isolated are psychrophilic strains of Escherichia (119), Aerobacter, Aeromonas (120, 121), Serratia (101, 122), Proteus (123), Chromobacterium (124), and Vibrio (125). Although facultative anaerobic psychrophiles have been recognized for some years (126, 127), obligate anaerobes capable of growing well at 0 were reported only recently (128); these bacteria were judged to be strains of Clostridium. Aerobic, spore-forming psychrophilic bacteria were once thought not to exist, but Larkin & Stokes (129) have reported isolation of psychro philic strains of Bacillus from soil, mud, and water. Spore formation and spore germination by these bacilli also took place at 0. There have been occasional reports of the isolation of psychrophilic strains of Corynebacterium, Lacto bacillus, and Micrococcus (79), but few of these reports include definitive data on the effect of low temperatures on growth. The commonest psychrophilic yeasts are members of the genus Candida (98, 99, 130), Strains of Cryptococcus (131, 132), Rhodotorula (106), and Torulopsis (106) are also frequently isolated; there are reports, too, of psy chrophilic strains of Debaryomyces and Pichia (106). Not so much is known about the distribution of the psychrophilic habit among fungal genera. The most informative account has come from Gunderson (133) who reported on moulds isolated from frozen food products. These organisms were selected on the basis of their ability to grow at 5, although the author states that, in general, fungi which grow at 5 are still able to grow at 0. Among the 113 identified isolates, there were representatives from 21 genera; Aureobasidium (= Pullularia) pullulans was the most frequently isolated species, followed by Botrytis cinerea, Phoma sp., and Geotrichum candidum. Gunderson also noted an association between the appearance of a psychrophilic mould and the na ture of the foodstuff; Chrysosporium pannorum appears to be associated with meat pies, while Phoma spp. are more prevalent on cherry fruit fillings. Even less has been reported on psychrophilic algae. Probably the best known are those that grow on the surface of snow and are a common sight in the polar regions and on mountains. These algae include species of Scotiella and Raphidonema, and Ancyclonema nordenskioldii, A. meridionale, Chlamy domonas nivolis, and C. ./lavo-virens (134). Many marine algae must be psy chrophilic, but few studies have been reported on the effects of low tempera tures on growth of these organisms. However, several strains of diatoms have been isolated from sea ice. including strains of Amphiprora, Biddulphia, Coscinodiscus, Eucampia, Fragilaria, Nitzschia, Pleurosigma, and Rhizoso lenia (107); these algae are almost certainly psychrophilic. The reasons for the uneven distribution of the psychrophilic habit among genera of microorganisms are frankly puzzling. The fact that psychrophilism in bacteria is most often found in Gram-negative organisms suggests a rela tionship between the ability to grow at 0 and those properties of the bacte rium which allow it to lose the crystal violet stain after washing with ethanol

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

110

FARRELL & ROSE

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

or acetone. Since there are reasons to believe that the ability to grow at 0 may be associated with certain properties of the cytoplasmic membrane, one is tempted to suggest that the possession of a double membrane in the cell envelopes of Gram-negative bacteria may predispose a bacterium to grow at 0. Clearly, however, there are many other factors involved, for not all Gram negative bacteria are psychrophilic, added to which psychrophilic yeasts ap parently do not have differently constructed cell envelopes as compared with mesophilic yeasts. Another property of bacteria which appears to be corre lated, at least to some extent, with the ability to grow at 0, is the presence of a high proportion of G + C residues in their DNA (135); data are not avail able to show whether a similar situation exists in the psychrophilic yeasts. Exactly what bearing such a compositional difference in DNA may haye on the ability to grow at 0 is not known. MICROB IAL METABOLISM AT SUBOPTIMUM TEMPERATURES Large numbers of microorganisms grow and reproduce in environments that are at temperatures many degrees below their optima for growth. The span between the optimum and the minimum temperatures for growth of a microorganism is usually around 20 to 35 (3); the span is invariably greater than that between the optimum and the maximum temperatures for growth. The fundamental question that needs to be answered regarding growth of microorganisms at suboptimum temperatures was stated quite succinctly by Foter & Rahn (136) in 1936 when they observed that, since growth and res piration consist of chemical reactions, they should, in theory, continue, al though at a greatly diminished rate, until the medium in which the micro organisms are suspended freezes solid. This behaviour is found with psy chrophiles but not with mesophiles, and several workers have attempted to discover the basis of the apparently aberrant behaviour of mesophiles. It has, moreover, often been assumed that the rate of individual metabolic pro cesses decreases at the same rate as the growth rate, as the temperature is de creased from the optimum. As shown below, there is now abundant evidence that this is not always a correct assumption, and the implications which these findings have in relation to the activities of microorganisms in natural environments are gradually coming to be appreciated.
PHYSIOLOGICAL ACTIVITIES

Pigment produ ct ion. Synthesis of the red pigment, prodigiosin, by strains of Serratia marcescens is favoured between 20 and 25 although the optimum temperature for growth of the bacterium is nearer 3r. The enzyme that catalyzes the final step in the biosynthesis of prodigiosin, the coupling of a monopyrrole with a bipyrrole to give the linear tripyrrole, prodigiosin, is abnormally temperature-sensitive (137) . Other examples of an increased pro duction of pigment by microorganisms at suboptimum temperatures are known. With some organisms, the effect has been traced to an influence of
-

TEMPERATURE ON MICROORGANISMS

111

temperature on enzyme synthesis rather than on enzyme activity as, for ex ample, with synthesis of the red pigment by the silk worm pathogen, B. cereus var. elesti which takes place at 15 but not at 28 (138). Flagella production.-Production of flagella is often markedly affected by temperatures within the range for growth of a microorganism. It is often favoured at low temperatures in the range and is absent at higher tempera tures; such examples include Bacillus inconstans (139), Bordetella bronchisep tica (140), E. coli (141), Pasteurella pseudo tuberc ulosis (142), Salmonella paratyphi B (143), Salmonella typhi (144), and certain psychrophilic bacteria (145). The opposite effect, namely, synthesis of flagella at high but not at low temperatures in the growth range, has been reported for other psychrophiIic bacteria (145), Salmonella paratyphi C (146) and Phytophthora infestans (147). Roberts & Doetsch (148) studied the effect of temperature on regener ation of flagella in bacteria that had been experimentally deflagellated, and showed that the thermophile, B. stearothermophilus 11330 resynthesized flagella at 20, a temperature below the minimum for the growth of the bacterium. In view of these data, the desirability of retaining flagella pro duction as a taxonomic criterion is clearly in question (149). Polysaccharide synthesis - S ynthesi s of polysaccharides, either capsular or intracellular, is often greater at suboptimum temperatures than at the optimum temperature for growth of a microorganism. This effect has been well documented in connexion with the production of extracellular dextrans by many Leuconostoc species and lactic acid bacteria; certain of these bacteria produce almost no dextran at 37, whereas, at 25, the cultures become ex tremely viscous because of the accumulation of an extracellular dextran. The effect, has been attributed to the formation by leuconostocs of a dextran sucrase that is rapidly inactivated at temperatures above about 30 (150). With some of the lactobacilli, higher temperatures are thought to prevent synthesis of the dextransucrase (151). Cursory observations in our own and in other laboratories have shown that several microorganisms accumulate extracellular polysaccharides when grown at temperatures near their mini mum for growth; unfortunately, no details have been published on these data. Tempest & Hunter (152) reported an increase from 3.3 per cent to 8.9 per cent in the proportion of dry weight accounted for as carbohydrate when the temperature for growth of a continuous culture of A. aerogenes was de creased from 35 to 25. However, these workers did not indicate whether the increase was attributable to the more extensive deposition of intracellular polysaccharide reserves or to an increase in the amount of cell wall poly saccharide in the bacteria. This tendency of certain microorganisms to accumulate greater quantities of polysaccharides at suboptimum temperatures may be of considerable im portance in relation to the survival of organisms in natural environments. The effect merits a more detailed examination by microbial physiologists. Fatty acid synthesis.-It has long been known that the composition of the
.

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

112

FARRELL & ROSE

lipids in poikilothermic organisms varies with the growth temperature [for a review, see (153)]. In general, organisms grown at lower temperatures in the growth range contain an increased proportion of unsaturated fatty acid re sidues in their lipids. This effect of temperature on lipid composition has been described in detail for mesophilic and psychrophilic strains of Candida (154) and Serratia (155, 156) and for E. coli (153). Kates & Baxter (154) reported an approximately 50 per cent increase in the content of double bonds in fatty acids from mesophilic and psychrophilic candidas grown at 10 as compared with 25. The alterations in fatty acid composition caused by the change in incubation temperature are more extensive in bacteria partly because of the greater diversity of fatty acids in these organisms. The main alterations caused by changing the incubation temperature for E. coli from 35 to 10 were increased proportions of hexadecenoic and octadecenoic acids (153). Strains of Candida grown at 25 and 10 had similar phospholipid and neutral lipid compositions; the decrease in temperature caused an increase in the con tents of linoleic acid at the expense of oleic acid in the mesophile C. lipolytica. Linolenic acid was detected in the psychrophiles grown at 10 but not in the mesophile grown at this temperature. This would appear to be a strain dif ference, as Candida utilis NCYC 316 contains linolenic acid even when grown at 30 (157). These reports also show that the composition of the medium can materially affect the fatty acid composition of organisms at any one tempera ture. In addition, E. coli harvested in the stationary, but not in the expo nential, phase of growth contained large amounts of cyclopropane fatty acid (153). C. lipolytica showed marked changes in the proportions of oleic and linoleic acids during the growth cycle at 10 or 25 (154); the fatty acid com position of the psychrophilic candidas, on the other hand, remained fairly constant during the growth cycle. Almost nothing is known of the biochemical basis of this effect of tem perature on the fatty acid composition of microorganisms. The possibility that the changes are caused by purely chemical factors has been discounted (154); nor is the fatty acid composition, at least with E. coli, a peculiar func tion of growth rate (153). It has been suggested that the fatty acid composi tion of an organism at a particular temperature is the result of differences in the rates of fatty acid synthesis and oxidative breakdown at that tempera ture (154). However, the most plausible explanation is that temperature di rectly affects the synthesis or activity, or both, of enzymes that catalyze re actions leading to the synthesis of unsaturated fatty acids. A report by Meyer & Bloch (158) indicates that growth temperature affects synthesis of the enzymes in Torulopsis ( Candida) utilis. Yeasts are suitable organisms for studying this effect, since the unsaturated acids are formed from the corresponding saturated acids. But, because of the effect of medium com position, and in all likelihood oxygen concentration too, these studies will need to be done in an oxystat. Solute uptak e.-Solutes are transported across the microbial cytoplasmic membrane by stereospecific proteins known as permeases. These proteins are
=

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

TEMPERATURE ON MICROORGANISMS

113

di5tinct from, but have many properties in common with, metabolic enzyme5 including temperature-dependence (159). Baxter & Gibbon5 (160) compared the temperature-dependence of a glucose permease, using glucosamine hydro chloride as a non-metabolizable analogue, and of growth and respiratory ac tivity in the mesophile, C. lipolytica and a psychrophilic candida. There was scarcely any solute uptake by C. lipolytica at 0 and 10, although the psy chrophile rapidly transported glucosamine even at 0. Baxter & Gibbons con cluded that the minimum temperature for growth of the mesophile was set by the temperature at which the solute-transport mechanisms were inactivated. This was the first suggestion for a physiological basis for the minimum tem perature for growth of a microorganism. Work in our own laboratory, with strains of Arthrobacter and C. utilis, gave further evidence for permease inac tivation in determining the minimum temperature for growth (17). There are several other reports showing the marked decrease in the rate of solute up take by microorganisms as the temperature is decreased below the optimum for growth. These include uric acid and xanthine uptake by C. utilis (161), monosaccharide uptake by Saccharomyces cerevisiae (162), uptake of inor ganic sulphate by Penicillium chrysogenum (163), and uptake of adenosine and inosine by E. coli (164). However, none of these reports has related these effects to the minimum temperature for growth of the particular mesophile. Possible explanations for the molecular basis of the effect of low tempera ture5 on 50lute uptake by microorgani5ms have been discussed at length by us in another review (3). Stated briefly, there are three basic mechanisms by which low temperature could affect solute uptake, and any one, or combina tion of these, might be instrumental in determining the minimum tempera ture for growth: these are: (a) inactivation of individual permease proteins at low temperatures as a result of low-temperature induced conformational changes which have been shown to occur in some proteins (165); (b) changes in the molecular architecture of the cytoplasmic membrane which prevent permease action; (c) a shortage of energy required for the active transport of solutes. As yet there is insufficient evidence to make a decision as to whether any of these mechanisms can be discounted. However, the possibility of the minimum temperature for growth being determined by the inactivation of individual permeases does not seem likely, mainly because this would mean that the minimum temperature of a microorganism would vary with the chemical composition of the medium, an observation which has not been re ported. A shortage of ATP for the active uptake of solutes also seems un likely, since endogenous respiration proceeds at temperatures below the minimum for growth of some microorganisms (17, 160); there is, nevertheless, the possibility that respiration at these temperatures is not coupled to ATP synthesis (166). This leaves (b) which, at the moment, seems the most likely explanation. However, the nature of the changes in membrane architecture that may be caused by low temperatures is unknown. It has been suggested that, at near-zero temperatures, the fatty acid side chains in membrane phospholipids solidify (167), and this could restrict the mobility of the per-

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

114

FARRELL & ROSE

mease proteins. Changes in membrane architecture may also be caused by a positional uncoupling of proteins and phospholipids, conceivably as a result of a change in the association between protein sulphydryl groups and fatty acid double bonds (168).

SYNTHESIS AND ACTIVITY

OF

ENZYMES

Total protein synthesis.-The rate of protein synthesis, in common with many other metabolic processes, decreases as the temperature is decreased below the optimum for growth of a microorganism. Goldstein and his col leagues (169) showed that protein synthesis takes place in E. coli at 0, and that the rate of synthesis at this temperature is about 350 times slower than that at 37. The amount of DNA in each microorganism, expressed as a per centage of the dry weight, does not alter significantly as the temperature is lowered (152). Data on the effect of temperature on RNA synthesis are meagre. Schaechter and his co-workers (170), using batch cultures of Sal monella typhimurium, showed that RNA content was not a function of growth rate when this rate was altered by changing the temperature over the range 37 to 25. However, Tempest & Hunter (152) reported that the RNA content of A. aerogenes, grown in a chemostat at a fixed rate under conditions of glucose limitation, increased as the temperature was decreased from 40 to 25. They assumed that the rate of protein synthesis, per unit of ribosome, was constant at a particular temperature. Lowering the temperature de creased ribosomal activity so that, in order to maintain a constant growth rate, the bacteria compensated by synthesizing additional ribosomal RNA. Synthesis of individual enzymes.- Rather more is known about the effects of temperature on the synthesis of individual enzymes [see Knox (171) for a review of the earlier literature]. More recently, there have been reports that synthesis of glutamate decarboxylase in a strain of E. coli is inducible at 37 but partly constitutive at 30 (172) while, in another strain of this bacterium, tryptophan induces synthesis of tryptophanase at 30 but not at tempera tures below 15 (173). Horiuchi & Novick (174) isolated a mutant strain of E. coli which synthesized ,a-galactosidase constitutively at 43.5 but needed the presence of an inducer to synthesize the enzyme at 14. Similar studies related to the effect of temperature on repression of enzyme synthesis have been reported (175, 176). Detailed studies on the effect of temperature on the kinetics of .8-galactosidase synthesis by E. coli ML 30 have come from Ingra ham's laboratory (37, 38). This group studied synthesis of the enzyme during exponential growth in a succinate-minimal medium over the range 10 to 43 for a strain with a constitutive ability to synthesize the enzyme, and in an inducible cryptic strain, induced maximally or submaximally with isopropyl thio-.8-galactoside. The differential rates of enzyme synthesis were identical for the constitutive strain and for the fully induced strain. The rates were constant from 20 to 43, and decreased proportionately with a decrease in temperature below 20. It was concluded that, in the absence of glucose re pression, the ability of E. coli to synthesize ,a-galactosidase decreases as the

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

TEMPERATURE ON MICROORGANISMS

115

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

temperature is decreased. These effects of temperature on enzyme synthesis have been explained by postulating that temperature affects synthesis of the repressor protein (38), that the repressor protein itself is thermolabile (176), or that the affinity of the repressor for the co-repressor is markedly influ enced by temperature (166). Reports by Szer & Ochoa (177) and Friedman & Weinstein (178) have drawn attention to another mechanism by which temperature may affect synthesis of individual enzymes, namely, by influencing the fidelity of the translation of mRNA during protein synthesis. Using a cell-free, amino acid incorporating system from the thermophile, B. stearothermophilus, it was found that the leucine-phenylalanine ambiguity, which is manifested when poly U is used as an artificial messenger, is greatly increased at low tempera tures in the growth range (178). It was suggested that this ability of tempera ture to affect the translation of mRNA may explain the behaviour of tem perature-sensitive mutants of phages and bacteria which synthesize enzymes at low but not at high temperatures in the growth range, since the low tem peratures, by inducing an ambiguity in the translation, may partially correct the effects of the mutated codon. TEMPERATURE-SENSITIVE MUTANTS There has been an increasing number of reports in recent years concerning the isolation of mutant strains of microorganisms, particularly bacteria, that have temperature responses different from those of the parent organism; these mutants are generally referred to as temperature-sensitive (ts) mu tants. There are two classes of ts mutants. The smaller class comprises mu tants that are able to grow at temperatures outside the range of the parent organism. For example, psychrophilic mutants have been obtained from mesophilic psuedomonads by the use of ultraviolet irradiation (179). How ever, the majority of ts mutants differ from their parent organisms in having a shorter temperature span for growth. Some of these mutants differ from the parents in that, at certain temperatures usually at the upper end of the growth range, they are auxotrophic for low-molecular weight compounds, synthesis of which is prevented at these temperatures in the mutant. Refer ence has already been made to some of these mutants and the use to which they have been put in studies on the regulation of enzyme synthesis (171174), and additional examples are given in the reviews by Ingraham (1) and Farrell & Rose (2), and in original papers (180, 181). There have been very few reports of mutants that are auxotrophic at low temperatures. However, Ingraham and his colleagues (182) isolated mutants of E. coli, all of which were cold-sensitive (cs) and had higher minimum temperatures for growth compared with the parent; certain of these mutants (K-II) had additional nutritional requirements at temperatures below 20. One of these mutants (K-II-27) required histidine at low temperatures, and was shown to syn thesize a phosphoribosyl-ATP pyrophosphorylase (the enzyme that catalyses the first reaction on the pathway to histidine synthesis), which is much more

116

FARRELL & ROSE

sensitive to feedback inhibition at 37 by histidine than is the enzyme from the parent. At temperatures below 37, the affinity for histidine increases with the enzymes from the mutant and the parent, but only with the mutant enzyme is it sufficient to cause a cessation of growth in the absence of exog enous histidine. But, probably the most interesting application of ts mutants has been in studies on the biosynthesis of macromolecules. Some ts mutants are unable to synthesize a macromolecule at temperatures at which the parent can and, since these macromolecules cannot be supplied exogenously, such mutations are conditionally lethal. There have been several reports of E. coli mutants that are unable to synthesize DNA at the restrictive temperature (183-187). Certain of the ts mutants described by Kohiyama and his colleagues (184, 186) were unable to initiate DNA replication at 42, although the activity of the DNA polymerase in these mutants was unaffected at the high tempera ture. It is hoped that data from experiments on these mutants will provide further information about the mechanisms that regulate DNA synthesis in bacteria. Information about the mechanisms that regulate RNA synthesis has also been obtained from studies on ts mutants (188-190). Eidlic & Neidhardt and their colleagues (188, 189) obtained several ts mutants of E. coli, using ethyl methane sulphonate as the mutagen. One of their mutants was derived from a strain with a stringent amino acid control of RNA synthesis. It did not synthesize RNA at the restrictive temperature (37), and was shown to possess a temperature-sensitive valyl-sRNA synthetase. The second mutant was derived from a strain with a relaxed control of RNA synthesis; this mu tant synthesized RNA at 37 and possessed a temperature-sensitive phenyl alanine-sRNA synthetase. The behaviour of these mutants suggests that amino acids permit RNA synthesis in stringent strains only after attachment or activation to sRNA, that relaxed strains can overproduce RNA without a complete complement of functioning aminoacyl-sRNA synthetases, and that the activity of these enzymes is obligatory for the biosynthesis of proteins. Yaniv and his co-workers (190) induced mutations in E. coli with N-methyl N' -nitro-N-nitrosoguanidine, and among the mutants unable to grow at 41 they obtained five with a temperature-sensitive, valine-activating enzyme. This preponderance of 'valyl' mutants is remarkable, but as yet entirely un explained.
LITERATURE CITED 1 . Ingraham, J. L., in The Bacteria, IV, 265-96. (Gunsalus, ' I. C., Stanier, R. Y., Eds., Academic Press, New York, 459 pp., 1962) 2. Farrell, J., Rose, A. H., A dflan. Appl. Microbial., 7, 335-78 (1965) 3. Farrell, J., Rose, A. H., in Thermobiol ogy, 147-218. (Rose, A. H., Ed., Academic Press, London, 653 pp., 1967) 4. Ryan, F. J., Kiritani, K, J. Gen. Microbial., 20, 644-53 (1959) 5. Arpai, J., Appl. Microbiol., 10, 297301 (1962) 6. Nakamura, M., Dawson, D. A., Appl. Microbiol., 10, 40-43 (1962)

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

TEMPERATURE ON MICROORGANISMS
7. Hansen, N.-H., Riemann, H., J. Bacteriol., 26, 314-33 (1963) Lindeberg, G., Lode, A., Can. J. MiCTobiol., 9, 523-30 (1963) Postgate, J. R., Hunter, J. R, J. Appl. Bacteriol., 26, 405-14 (1963) Hagen, P.-O., Kushner, D. J., Gib bons, N. E., Can. J. Microbiol., 10, 813-22 (1964) Strange, R. E., Shon, M., J. Gen. Microbiol., 34, 99- 1 14 (1964) Ashwood-Smith, M. J., Bridges, B. A., Munson, R 1., Science, 149, 1 103-5 (1965) Dimmick, R L., J. Bacterial. , 89, 79198 (1965) Evison, L. M., Rose, A. H., J. Gen. Microbiol., 40, 349-64 (1965) M azur, P., Ann. N. Y. A cad. Sci., 125, 658-74 (1965) Packer, E. L., I ngraham , J. L., Scher, S., J. Bacteriol., 89, 7 1 8-24 (1965) Rose, A. H., Evison, L. M., J. Gen. Microbial., 38, 1 3 1 -4 1 (1 9 65 ) MacLeod, R A.. Smith, L. D. H., Gelinas, R, Can. J. Microbial., 12, 61-71 (1966) Mazur, P., in Cryobiology, 214-315. (M eryman, H. T., Ed . , Academic Press, New York, 775 pp., 1966) Long , S. K., Williams, O. B., J. Bac teriol., 79, 629-37 (1960) Curran, H. R, Pallansch, M. J., J. Bacterial., 86, 91 1-18 (1963) Knaysi , G., J. Bacteriol., 87, 619-22 (1965) Keynan, A., Issahary-Brand, G., Evenchik, Z., in Spores, 111, 180-87. (Campbell, L. L., Halvorson, H. 0., Eds., Am. Soc. Microbiol., Ann Arbor, Mich., 1965) Murrell, W. G., Warth , A. D., in Spores, III, 1-24. (Campbell, L. L., Halvorson, H. 0., Eds., Am. Soc. Microbiol., Ann Arbor, Mich., 1965) Yeas, N., Exptl. Cell Res., 10, 746 (1956) Gutz, H., Naturwissenschaften, 44, 545
Appl.

117

8.
9.

33. Borgstrom, G. A., in Low Temperature

Agr., Albany, Cal., 40-61 (1960)

Microbiology, 197-251. (Campbell Soup Co., Camden, New Jersey,

322 pp., 1962)

10. 11. 12.

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

13. 14. 15.

1 6.

34. Peterson, A. C., in Low Temperature Microbiology, 1 5 7-95. (Campbell Soup Co., Camden, New Jersey, 322 pp., 1962) 35. Michener, H. D., Elliott, R P., A dvan. Food Res., IS, 349-96 ( 1 9 65) 36. Silverman, G. 1., Goldblith, S. A., A dvan. Appl. Microbiol., 7, 305-34 (1965) 37. Ng, H., Ingraham, J. L., Marr, A. G., J. Bacteriol., 84, 331-39 (1962) 38. Marr, A. G., Ingraham, J. L., Squires, C. L., J. Bacterial., 87, 356-62 (1964) 39. Stanley, S. 0., Rose, A. H., J. Gen.
Microbial. (In press)

1 7. 18. 19. 20. 21. 22.

23.

40. Nakae, T., J. Bacterial., 91, 1 730-35

(1966)

24.

25. 26.

(1957)

27. Ogur, M., Ogur, S., St. John, R., Genetics, 45, 1 89-98 (1960) 28. Zamenhof, S., Proc. Natl. A cad. Sci. U.S., 46, 101-5 (1960) 29. Chiasson, L. P., Zamenhof, S., Can. J. Microbiol., 12, 43-46 (1966) 30. McNelly-Ingle, C. A., Lamb, B. C., Frost, L. C., Genet. Res., Cambridge, 7, 1 69-83 (1966) 3 1 . Borgstrom, G. A., A dvan. Food Res., 6, 1 63-230 (1955) 32. Elliott, R P., M ichener, H. D., Conf. Frozen Food Qual ity, U.S. Dept.

41. Landman, O. E., Bausum, H. T., Matney, T. S., J. Bacterial. , 83, 463-69 (1962) 42. Oppenheimer, C. H., Drost-Hansen, W., J. Bacterial. , 80, 2 1-24 (1960) 43. Cannefax, G. R., J. Bacteriol. , 83, 70810 (1962) 44. Elliott, R P., Heiniger, P. K., Appl. Microbiol., 13, 73-76 (1965) 45. Dimmick, R. L., Appl. Microbiol., 1 3, 846-50 (1965) 46. Battley, E. H., Antonie van Leeuwen hoek, J. Microbiol. Serol., 30, 81-96 (1964) 47. Fries, N., Physiol. Plantarum, 16, 41522 (1963) 48. Halldal, P., French, C. S., Plant Physiol ., 33, 249-52 (1958) 49. Fogg, G. E., Bacterial. Rev., 20, 148-65 (1961) 50. Kempner, E. S., Science, 142, 1 3 1 8 (1963) 51. Gaughran, E. R L., Bacteriol. Rev., 1 1, 189-225 (1947) 52. Allen, M . B., Bacterial. Rev., 17, 12573 (1953) 53. Fields, M. L., Appl. Microbial., 1 1 , 100-4 (1963) 54. Edwards, J. L., Jr., Busta, F. F., Speck, M. L., Appl. Microbiol. , 1 3, 851-57 (1965) 55. Edwards, J. L., Jr., Busta, F. F., Speck, M. L., Appl. Microbial., 13, 858-64 (1965) 56. Yokoya, F., York, G. K., Appl. Micro bial., 13, 993-99 (1965) 57. McDonald, W. C., Matney, T. S., J. Bacteriol. , 85, 2 1 8-20 (1963)

1 18

FARRELL & ROSE

84. Stokes, J. L., Redmond, M. L., Appl. Microbio!., 14, 74-78 (1966) 85. Peterson, A. C., Gunderson, M . F., Appl. Microbiol., 8, 98-104 (1960) 86. Ohye, D . F., Scott, W. J., A ustralian J. Bioi. Sci., 10, 85-94 (1957) 87. Segner, W. P., Schmidt, C. F., Boltz, J. K., Appl. Microbiol., 14, 49-54 (1966) 88. Cann, D. C., Wilson, B. B., Hobbs, G., Shewan, J. M., J. Appl. Bacteriol., 28, 431-36 (1965) 89. Ekelof, E., W. Erg. der Schwed. Sud polar Expedition 1901-1 903, 4. (Stockholm, 1908) 90. Tsiklinsky, M., Flore Microbienne, ExpUition A ntarctic Francaise, 1903-1905. (Masson et Cie., Paris, 1908) 9 1 . Gazert, H., Deut. Sudpolar-Expedition, 1901-1903, 7, 268-96. (1912) 92. Pirie, J. R R, Notes on A ntarctic Microbiology. (Edinburgh, 1912) 93. McLean, A. L., Nature, 102, 35-39 (1918) 94. Darling, C. A., Siple, P. A., J. Bac teriol., 42, 83-98 (1941) 95. Bunt, J. S., A ustralian J. Marine Freshwater Res., 6, 60-65 (1955) 96. Bunt, J. S., Rovira, A. D., J. Soil Sci., 6, 1 1 9-26 (1955) 97. Boyd, W. L., Boyd, J. W., J. Bacteriol., 85, 1 1 2 1-23 (1963) 98. Straka, R. P., Stokes, J. L., J. Bac terial., 80, 622-25 (1960) 99. Di Menna, M . E., J. Gen. Microbiol. , 2 3 , 295-300 (1960) 100. Sinclair, N. A., Stokes, J. L., Can. J. Microbial. 1 1, 259-69 (1965) 101. Stanley, S. 0., Rose, A. H., Proc. Roy. Soc., Ser. B. (In press) 102. Zobell, C. E., in Law Temperature Microbiology, 107-32. (Campbell Soup Co., Camden, New Jersey, 322 pp., 1962) 103. Morita, R. V., Oceanogr. Marine Bioi. Ann. Rev., 4, 105-21 (1966) 104. Kriss, A. E., Marine Microbiology (Deep Sea). (Trans!. by Shewan, J.

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

58. Waksman, S. A., Corke, C. T., J. Bacteriol., 66, 377-78 (1953) 59. Tendler, M. D., Burkholder, P. R., Appl. Microbiol., 9, 394-99 (1961) 60. Erikson, D., J. Gen. Microbio!., 13, 127-35 (1955) 61. Lindt, W., Palhol. Pharmakol., 21, 269-98 (1 886) 62. Cooney, D. G., Emerson, R., Thermo philic Fungi. (W. H. Freeman & Co., San Francisco, 188 pp., 1964) 63. Koffler, H., Bacteriol. Rev., 21, 227-40 (1957) 64. Marsh, C., Militzer, W., A rch. Bio chem. Biaphys., 36, 269-75 (1952) 65. Marsh, c., M ili tzer, W., A rch. Bio chem. Biophys., 60, 439-51 (1956) 66. Mathemeier, P. F., Morita, R. Y., J. Bacteriol., 88, 1661-66 (1964) 67. Militzer, W., Tuttle, L. C., Georgi, C. E., Arch. Biochem. Biaphys., 31, 416-23 (1951) 68. Thompson, P. J., Thompson, T. S., J. Bacteriol., 84, 694-700 (1962) 69. Akagi, J. M., Campbell, L. L., J. Bacterial., 82, 927-32 (1961) 70. Akagi, J. M . , Campbell, L. L., J. Bacteriol., 84, 1 194-1201 (1962) 7 1 . Campbell , L. L., Postgate, J. R., Bacterial. Rev., 29, 359-98 (1965) 72. Manning, G. B., Campbell, L. L., J. Bioi. Chem., 236, 2952-57 (1961) 73. Abram, D., Koffler, H., Backrial. Proc., 30 (1965) 74. Marmur, J., Biochim. BioPhys. A cta 38, 342-43 (1960) 75. Teece, G., Toschi, G., Nuovo Cimento, Suppl., 18, 207- 1 1 (1960) 76. Mangiatini, M. T., Teece, G., Toschi, G., Nuovo Cimento, Suppl., 25, 4550 (1962) 77. Saunders, G. F., Campbell, L. L., J. Bacteriol., 91, 332-39 (1966) 78. Ingraham, J. L., Stokes, J. L., Bac terial. Rev., 23, 97-108 (1959) 79. Witter, L. D., J. Dairy Sci., 44, 9831015 (1961)

80. Jacobs, M. B., Gerstein, M. J., Walter, W. G., Eds., Dictionary of Microbiology. (Van Nostrand, New
York, 276 pp., 1957) 81. Burrows, W., Textbook o Microbiology. f 954 pp., 1959)

CW. B. Saunders Co., Philadelphia,

82. Stokes, ]. L., in Recent Progress in Microbiology, VIII, 1 87-92 (Gib bons, N. E., Ed., University of Toronto Press, Toronto, 721 pp., 1 963) 83. Eddy, B. P., J. Appl. Bacteriol ., 23, 189-90 (1960)

M . , Kabata, Z., Oliver & Boyd, Edinburgh, 536 pp., 1963) 105. Wood, E. J. F., Marine Microbial Ecology. (Chapman & Hall, Lon don, 243 pp., 1965) 106. Norkrans, B., A rch. Mikrobiol., 54,
374-92 (1966) 107. Bunt, J. S., Wood, E. J. F., Nature, 199, 1 254-55 (1963) 108. Frank, R A., J. Bacteriol., 84, 68-71 (1962) 109. Stanier, R. Y., Palleroni, N. J.,

TEMPERATURE ON MICROORGANISMS
Doudoroff, M., J. Gen. Microbial.,
43, 159-2 7 1 (1966)

119
J.
Bacterial.,

136. Fater, M. J"

Rahn, 0., 32, 485-97 (1936)

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

1 10. Hucker, G. J., Food Technol. , 8, 79-84 (1954) 1 1 1 . Schultze, W. D., Olson, J. C., Jr., J. Dairy Sci., 40, 602 (1957) 1 12 . . Schultze, W. D., Olson, J. C., Jr., J. Dairy Sci., 43, 346-50 (1960) 1 13. Liston, J., J. Gen. Microbiol., 16, 20516 (1957) 1 14. Jones, G. E., Thomas, S. B., Proc. Soc. Appl. Bacteriol., 13 (1950) 1 15. Davis, P. A., Babel, F. ]., J. Dairy Sci., 37, 1 76-84 (1954) 1 1 6. Abd-el-Malek, Y., Gibson, T., Dairy Sci., 19, 294-302 (1952) 117. Andrey, J., Frazier, W. C., J. Dairy Sci., 42, 1 7 8 1-84 (1959) 1 1 8 . Roth, N. G., Wheaton, R. B., J. Bacterial., 83, 551-55 (1962) 1 19. Haines, R. B., J. Hyg., 34, 2 7 7-82 (1934) 120. Brown, A. D., Weidemann, J. F., J. Appl. Bacteriol ., 2 1 , 1 1- 1 7 (1958) 1 2 1 . Nagel, C. W., Simpson, K. L., Ng, H.,
Food Technol., 14, 2 1-32 (1960)

137. Williams, R. P., Goldschmidt, M. E., Gott, C. L., Biochem. Biaphys. Res. Commun., 19, 1 77-81 (1965) 138. Uffen, R. L., Canale-Parola, E., Can. J. Microbial., 12, 590-93 (1966) 139. Braun, H., Lowenstein, P., Z . Bak terial. Parasitenk., 91, 1-56 (1923) 140. Lacey, B. W., J. Hyg., 58, 5 7-93 (1953)

141. Morrison, R. B., McCapra, J., Na ture, 192, 774-76 (1961) 142. Preston, N. W., Maitland, H. B., J. Gen. Microbiol . , 7, 1 1 7-28 (1952) 143. Jordon, E. 0., Caldwell, M. E., Reiter, D., J. Bacteriol. , 27, 1 65-74

Vaughn, R. H., Stewart, G. E.,

122. Thomas, S. B., Griffiths, ]. M., Foulkes, J. B.o Dairy Eng., 77, 438-40 (1960) 123. Campbell, J. J, R., Reed, G. B., Can. J. Res., 25, 280-83 (1947) 124. Sneath, P. H. A., J. Gen. Microbiol., IS, 70-98 (1956) 125. Colwell, R. R., Morita, R. Y., J. Bacteriol., 86, 831,...3 7 (1964) 126. Muller, M., Arch. Hyg., 47, 1 2 7-93 (1903) 1 2 7. Upadhyay, J., Stokes, J. L., J. Bac teriol. , 83, 2 70-75 (1962) 128. Sinclair, N. A., Stokes, J. L., J. Bac teriol., 87, 562-65 (1964) 129. Larkin, J. M., Stokes, J. L., J. Bac teriol., 91, 1667-71 (1966) 130. Lawrence, N. L., Wilson, D. C.,
7 , 7-1 1 (1959)

Pederson, C. S., Appl. Microbiol. ,

1 3 1 . Hagen, P.-O., Rose, A. H., Can. J. Microbiol., 7, 287-94 (1961) 132. Hagen, P.-O., Rose, A. H., J. Gen. Microbiol., 27, 89-99 (1962) 133. Gunderson, M. F., in Low Temperature Microbiology, 299-3 12. (Campbell

(1934) 144. Felix, A., Bhatnagar, S. S., Pitt, R. M., Brit. J. Exptl. Pathol. , 15, 346-54 (1934) 145. Schubert, 0., Schubert, R., Z. Hyg. Inf ektionskrank., 136, 639-42 (1953) 146. Quadling, C., J. Gen. Microbiol., 18, 22 7-37 (1958) 147. Ferris, V. R., Science, 120, 71-73 ( 1954) 148. Roberts, F. F., Doetsch, R. N., J. Bacterial., 91. 414,-21 (1966) 149. Orr, D. A., Taylor, M. M . , Nature, 2 08, 1017 (1965) 150. Neely, W. B., Advan. Carbohydrate Chem., 15, 341-69 (1960) 1 5 1 . Dunican, L. K., Seeley, H. W., J. Bacteriol., 86, 1079-83 (1963) 152. Tempest, D. W., Hunter, J. R., J. Gen. Microbial ., 41, 267-73 (1965) 153. Marr, A. G., Ingraham, J. L., J. Bac terial., 84, 1 260-67 (1962) 154. Kates, M., Baxter, R. M., Can. J. Biochem. Physiol., 40, 1 2 1 3-27 (1962) 155. Kates, M., Hagen, P.-O., Can. J. Biochem., 42, 481-88 (1964) 156. Bishop, D. G., Still, J. L., J. Lipid Res., 4, 87-93 (1963) 157. McMurrough, I . (Personal communi 158. Meyer, F., Bloch, K., Biochim. Bio phys. A cta, 77, 671-73 (1963) 159. Kepes, A., Cohen, G. N., in The Bac teria, IV, 1 79-221 . (Gunsalus, 1. C.,

cation)

Soup Co., Camden, New Jersey, 322 pp., 1962) 134. Marre, E., in Physiology and Biochem istry of A lgae, 541-50. (Lewin, R. A., Ed., Academic Press, New York, 840 pp., 1962) 1 3 5 . Hill, L. R., J. Gen. Microbiol. 44, 419,

Stanier, R. Y., Eds., Academic Press, New York, 459 pp., 1962) 160. Baxter, R. M., Gibbons, N. E., Can. J. Microbiol., 8, 5 1 1- 1 7 (1962) 1 6 1 . Quetsch, M. F., Danforth, W. F., J.
(1964) 162. van Steveninck, J., Biochim. Biophys., Acta, 126, 154-62 (1966)
Cellular Compo Physiol. , 64, 1 23-29

37 (1966)

1 20

FARRELL & ROSE

1 7 7 . Szer, W., Ochoa, ,5., J. Mol. Bioi., 8, 823-34 (1964) 178. Friedman, S. M . , Weinstein, I. B.,
Proc. Natl. A cad. Sci. U.S., 52,

163. Yamamoto, L., Segel, I. H., Arch. Bio chem. Biophys., 1 14, 523-29 (1966) 164. Peterson, R. N., Koch, A. L., Bio
chim. Biophys. A cta, 126, 129-40

165.

166. 167. 168.

Annu. Rev. Microbiol. 1967.21:101-120. Downloaded from www.annualreviews.org by University of Saskatchewan on 11/10/11. For personal use only.

169.

1 70.

1 71.

1 72. 1 73 . 1 74.

175. 176.

(1966) Brandts, J. F., in Thermobiology, 2572. (Rose, A. H., Ed., Academic Press, London, 653 pp., 1967) Ingraham, ]. L. (Personal communica tion) Byrne, P., Chapman, D., Nature, 202, 987-88 (1964) Robinson, J. D., Nature, 212, 199-200 (1966) Goldstein, A., Goldstein, D. B., Lowney, L. I., J. Mol. Bioi., 9, 2 1 335 (1964) Schaechter, M . , Maalj6e, 0., Kjeld gaard, N. 0., J. Gen. Microbial , 19, 592-606 (1958) Knox, R., in Adaptation in Micro organisms, Symp. Soc. Gen. Micro bial., 184200 (1953) Halpern, Y. S., B iochem. Biophys. Res. Commun., 6, 33-37 (1961) Ng, H., Gartner, T. K., J. Bacteriol., 85, 245-46 (1963) Horiuchi, T., Novick, A., Cold Spring Harbor Symp. Quant. Bioi., 26, 247-48 (1961) Gallant, J., Stapleton, R., Pro. Nail. A cad. Sci. U.S., 50, 348 : (1963) Udaka, M., Horiuchi, T Eiochem. BioPhys. Res. Commun., 19, 156-60 (1965)
. ..

179.

180.

181. 182.

183.

988-95 (1964) Azuma, Y., Newton, S. B., Witter, L. D., J. Dairy Sci., 45, 1529-30 (1962) Maas, W. K., Davis, B. D., ProG. Natl. A cad. Sci. U.S., 38, 785-97 (1952) Horowitz, N. H., Fling, M., Genetics, 38, 360-74 (1953) O'Donovan, G. A., Kearney, C. L., Ingraham, J. L., J. Bacteriol., 90, 6 1 1 -1 6 (1965) Jacob, F., Brenner, S., Cuzin, F.,
Cold Spring Harbor Symp. Quant. Bioi., 28, 329-36 (1963)

184. Kohiyama, M., Lanfrom, H., Brenner, S., Jacob, F., Campt. Rend., 257, 1979-81 (1963) 185. Bonhoeffer, F., Schaller, H., Biochem. Biophys. Res. Commun., 20, 93-97 (1965) 186. Kohiyama, M., Cousin, D., Ryter, A., Jacob, F., Ann. Inst. Pasteur, 1 10, 465-68 (1966) 187. Kjeldgaard, N. 0., A dvan. Microbial Physiol., 1, 39-95 (1967) 1 88. Eidlic, L., Neidhardt, F. C., J. Bac teriol. , 89, 706-11 (1965) 1 89. Bock, A., Faiman, L. E., Neidhardt, F. C., J. Bacterial., 92, 1076-82 (1966) 190. Yaniv, M., Jacob, F., Gros, F., Bull. S'ilc. Chim. Bioi., 47, 1609-13 (1965)