Isolation of Ice-Nucleating Active Bacteria from the Freeze-Tolerant Frog, Rana sylvatica

Lee, Marcia R.; Lee, Richard; Strong‐Gunderson, Janet M.; Minges, Stacey R.

doi:10.1006/cryo.1995.1036

Cited by 50 publications

(26 citation statements)

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“…The strains were grown at 24°C on 1/10 TSA and 1/10 TSA supplemented with 5% glycerol, which has been shown to optimize ice nucleation activity (INA) in many bacteria [23,24]. The strains were grown at 24°C on 1/10 TSA and 1/10 TSA supplemented with 5% glycerol, which has been shown to optimize ice nucleation activity (INA) in many bacteria [23,24].…”

Section: Ice Nucleation Activitymentioning

confidence: 99%

“…Ice nucleation studies were undertaken to determine the effect of temperature on ice nucleation activity in seven permafrost strains. The strains were grown at 24°C on 1/10 TSA and 1/10 TSA supplemented with 5% glycerol, which has been shown to optimize ice nucleation activity (INA) in many bacteria [23,24]. INA was measured by the freezing drop method [25].…”

Section: Ice Nucleation Activitymentioning

confidence: 99%

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Characterization of potential stress responses in ancient Siberian permafrost psychroactive bacteria

Ponder

Gilmour

Bergholz

et al. 2005

FEMS Microbiology Ecology

101

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Past studies of cold-acclimated bacteria have focused primarily on organisms not capable of sub-zero growth. Siberian permafrost isolates Exiguobacterium sp. 255-15 and Psychrobacter sp. 273-4, which grow at subzero temperatures, were used to study cold-acclimated physiology. Changes in membrane composition and exopolysaccharides were defined as a function of growth at 24, 4 and -2.5 degrees C in the presence and absence of 5% NaCl. As expected, there was a decrease in fatty acid saturation and chain length at the colder temperatures and a further decrease in the degree of saturation at higher osmolarity. A shift in carbon source utilization and antibiotic resistance occurred at 4 versus 24 degrees C growth, perhaps due to changes in the membrane transport. Some carbon substrates were used uniquely at 4 degrees C and, in general, increased antibiotic sensitivity was observed at 4 degrees C. All the permafrost strains tested were resistant to long-term freezing (1 year) and were not particularly unique in their UVC tolerance. Most of the tested isolates had moderate ice nucleation activity, and particularly interesting was the fact that the Gram-positive Exiguobacterium showed some soluble ice nucleation activity. In general the features measured suggest that the Siberian organisms have adapted to the conditions of long-term freezing at least for the temperatures of the Kolyma region which are -10 to -12 degrees C where intracellular water is likely not frozen.

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Section: Ice Nucleation Activitymentioning

confidence: 99%

Section: Ice Nucleation Activitymentioning

confidence: 99%

Characterization of potential stress responses in ancient Siberian permafrost psychroactive bacteria

Ponder

Gilmour

Bergholz

et al. 2005

FEMS Microbiology Ecology

101

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“…Icenucleating active bacterial strains capable of initiating freezing of aqueous solutions at temperatures greater than or equal to Ϫ4.0ЊC were chosen for this study (Table 1). Bacterial strains used were frog-derived Pseudomonas fluorescens F26-4C (Lee et al 1995); insect-derived P. fluorescens 88 Ð335, P. putida Hr6 Ð1, and P. syringae Hr6 Ð3B (M.R.L., unpublished data); and epiphytic P. syringae cit 7 (provided courtesy of S. E. Lindow, University of California, Berkeley). Bacteria were identiÞed using API 20E biochemical tests (BioMerieux Vitek, Hazelwood, MO) and conÞrmed by Analytab Products, Plainview, NY.…”

Section: Methodsmentioning

confidence: 99%

Identification of Ice-Nucleating ActivePseudomonas fluorescensStrains for Biological Control of Overwintering Colorado Potato Beetles (Coleoptera: Chrysomelidae)

Castrillo

Lee

et al. 2000

J Econ Entomol

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Laboratory studies were conducted to identify ice-nucleating active bacterial strains able to elevate the supercooling point, the temperature at which freezing is initiated in body fluids, of Colorado potato beetles, Leptinotarsa decemlineata (Say), and to persist in their gut. Adult beetles fed ice-nucleating active strains of Pseudomonas fluorescens, P. putida, or P. syringae at 10(6) or 10(3) bacterial cells per beetle had significantly elevated supercooling points, from -4.5 to -5.7 degrees C and from -5.2 to -6.6 degrees C, respectively, immediately after ingestion. In contrast, mean supercooling point of untreated control beetles was -9.2 degrees C. When sampled at 2 and 12 wk after ingestion, only beetles fed P. fluorescens F26-4C and 88-335 still had significantly elevated supercooling points, indicating that these strains of bacteria were retained. Furthermore, beetle supercooling points were comparable to those observed immediately after ingestion, suggesting that beetle gut conditions were favorable not only for colonization but also for expression of ice-nucleating activity by these two strains. The results obtained from exposure to a single, low dose of either bacterial strain also show that a minimum amount of inoculum is sufficient for establishment of the bacterium in the gut. Persistence of these bacteria in Colorado potato beetles long after ingestion was also confirmed using a polymerase chain reaction technique that detected ice-nucleating active bacteria by virtue of their ina genes. Application of these ice-nucleating active bacteria to elevate the supercooling point of this freeze-intolerant insect pest could significantly reduce their winter survival, thereby reducing local populations and, consequently, crop damage.

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“…Most of ice-nucleating-active (INA) bacteria are associated with plants (Lindow et al 1978;Lindermann et al 1982;Loper and Lindow 1994;Waturangi et al 2008) or animals (Lee et al 1995;Worland and Block 1999). Other microorganisms, including several genera of fungi (Pouleur et al 1992) and lichens (Kieft 1988) have been reported in their ability to induce ice formation.…”

mentioning

confidence: 99%

“…Isolate J83, which is closest related to P. fluorescens strain IAM12022, belonging to the gammasubdivison. P. fluorescens has been reported to belong to the bacteria which are able to catalyze ice formation (Lee et al 1995) while isolate J43 falls into the â-subdivison. This isolate is closely related to P. mephitica ATCC 33665T or J. lividum DSM 1522T.…”

mentioning

confidence: 99%

Free-Living Ice-Nucleating Active Bacteria from High Mountain Lake Habitats

Yuhana¹,

Hanselmann²

2008

Microbiol Indones

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We collected the culturable heterotrophic bacteria from oligotrophic high mountain lake habitats and tested their capability to induce ice formation. Direct plating was carried out using low-nutrient medium at a temperature of between 3 and 4°C. As many as 84 isolates were recovered from glacial ice and natural biofilm growing on granite rocks surface. Six out of 84 isolates were capable of expressing the ice-nucleation phenotype. After autoclaving the cell suspension at 121°C for 15 min, isolate J78 was still able to retain the ability for ice formation. Heat-stable ice nuclei produced by ice-nucleating active bacteria have potential applications in biotechnology. Characterization of INA bacteria was performed employing live-dead Gram staining and molecular methods. Universal primers for Bacteria (S-D-Bact-0008-b-S-20 and S-D-Bact-1524-a-A-18) were used for PCR to amplify almost the full length of the 16S rRNA genes of selected INA isolates. Restriction fragment length polymorphism analysis resulted in 2 unique patterns, as represented by J43 and J83, respectively. Based on DNA sequencing of 16S rRNA gene, isolate J43 (GeneBank accession no. AJ864852) was closely related to Pseudomonas mephitica (99.2% sequence similarity) and Janthinobacterium lividum (99% similarity), whereas isolate J83 (GeneBank accession no. AJ864859) showed 100% sequence identity to Pseudomonas fluorescens.

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Isolation of Ice-Nucleating Active Bacteria from the Freeze-Tolerant Frog, Rana sylvatica

Cited by 50 publications

References 0 publications

Characterization of potential stress responses in ancient Siberian permafrost psychroactive bacteria

Characterization of potential stress responses in ancient Siberian permafrost psychroactive bacteria

Identification of Ice-Nucleating ActivePseudomonas fluorescensStrains for Biological Control of Overwintering Colorado Potato Beetles (Coleoptera: Chrysomelidae)

Free-Living Ice-Nucleating Active Bacteria from High Mountain Lake Habitats

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