Spatial V= ial composition of water can be the difference between life or any organisms. Microbes in lakes can reproduce and spread to of the water cycle. Ice nucleating Pseudomonads are microbes en shown to be in all parts of the water cycle and damage plants, domonads conduct ice nueleation by inducing freezing of molecules. Through evaporation and rainfall, ads can disperse anywhere. When bacterial ice nucleation occurs t tissue, ice crystals form within the tissue and damages the plant. microbial composition, two Oklahoma lakes: the urban lake-- r and the rural lake-Crowder Lake, were examined. In each lake, les were collected from three specific locations in summer and aths. Samples were inoculated on 10% tryptic soy agar to otal bacterial load and King's B agar supplemented with boric neximide, and cephalexin to isolate Pseudomonads. At Lake eastern shore had significantly larger bacterial concentration in s compared to other locations. At Crowder Lake, the center of the nificantly lower bacterial concentration in both seasons compared ations. Seasonally, there was lower bacterial concentration in the ake Hefner compared to the summer, while Crowder Lake had erial concentration year-round. Lake Hefner had higher bacterial North to Weatherford miles water Figure 2. Sampling sites Crowder Lake are mar- stars (Yellow= Near Do- Center of Lake, Green Shore). Pseudomonad conc did not differ betwe sites; however, total load was significanth the center of

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Abstract
Spatial Va
The microbial composition of water can be the difference between life or
death for many organisms. Microbes in lakes can reproduce and spread to
other parts of the water eycle. Ice nucleating Pseudomonads are microbes
that have been shown to be in all parts of the water cycle and damage plants,
These Pseudomonads conduct ice nucleation by inducing freezing of
supercooled
Pseudomonads can disperse anywhere. When bacterial ice nucleation occurs
around plant tissue, ice crystals form within the tissue and damages the plant.
To examine microbial composition, two Oklahoma lakes: the urban lake-
Lake Hefner and the rural lake-Crowder Lake, were examined. In each lake,
water samples were collected from three specific locations in summer and
winter months. Samples were inoculated on 10% tryptic soy agar to
determine total bacterial load and King's B agar supplemented with borie
acid, cycloheximide, and cephalexin to isolate Pseudomonads. At Lake
Hefner, the eastern shore had significantly larger bacterial concentration in
both seasons compared to other locations. At Crowder Lake, the center of the
lake had significantly lower bacterial concentration in both seasons compared
to other locations. Seasonally, there was lower bacterial concentration in the
winter at Lake Hefner compared to the summer, while Crowder Lake had
similar bacterial concentration year-round. Lake Hefner had higher bacterial
concentration compared to Crowder Lake. Pseudomonads isolated from both
seasons and lakes were tested for ice nucleation activity by inoculating in
super cooled water and detecting ice formation. Twenty-two ice nucleators
were detected at Crowder Lake and three ice nucleators were detected at
North to
Weatherford
2 miles
water
molecules. Through evaporation and rainfall,
Figure 2. Sampling sites
Crowder Lake are marl-
stars (Yellow = Near Do-
Center of Lake, Green=
Shore).
• Pseudomonad
conc
did not differ betwe
sites; however, total
load was significant
the
center
of
compared to the doc
(Fig. 3).
Lake Hefner in the summer, and none were detected in the winter. The
seasonal differences in the concentration of ice nucleating bacteria were
surprising because other studies have shown that cooler environments select
for increased ice nucleating bacteria.
Spatial V
Ice Nucleating Pseudomonads
Herintal
yrimpee
yrngan deud
·pringe un
erosos om wid
and cutivaed pants
Lake
snow
waters
Hefner -
Figure 4. Sampling sit
Lake Hefner are marl
stars (Orange = South
Shore, Purple = Near
Blue = Eastern Shore
%3D
%3D
Ennhment and diversficaon
-ayoystem
Figure 1. Movement of Pseudomonas syringae
throughout the water cycle (Morris et al., 2008).
Lake Hefner
Pseudomonads are bacteria that inhabit soils and are pathogens of
plants and animals. Ice nucleating Pseudomonads (hereafter referred to
as Ice+) are bacteria that can freeze supercooled water in their vicinity
at a higher temperature than freezing would occur in their absence.
Icet strains have been detected in all portions of the water cycle and
likely spread via precipitation (Fig. 1) (Morris et al., 2008). Due to the
enrichment of Ice+ strains in clouds, ice nucleation activity is
hypothesized to allow Pseudomonads to move via clouds in association
with aerosolized water. This can be problematic because Ice+ strains
can be transmitted to agricultural plants through the water cycle where
they cause disease (Morris et al., 2008), Furthermore, Ice+ strains can
cause frost injury via ice nucleation of water in plant tissues (Lindow et
al., 1982). Ice+ bacteria have been detected on multiple continents, in
all forms of water, and in all seasons.
(A) 8000
b
6000
4000
2000
Bacterial Loac
Figure 6. Seasonal b-
Hefner (A) and Crowe
Fig. 3. Values indicat
value <0.05).
Encmant nd
gcuttural
CFU/ml E
Transcribed Image Text:Abstract Spatial Va The microbial composition of water can be the difference between life or death for many organisms. Microbes in lakes can reproduce and spread to other parts of the water eycle. Ice nucleating Pseudomonads are microbes that have been shown to be in all parts of the water cycle and damage plants, These Pseudomonads conduct ice nucleation by inducing freezing of supercooled Pseudomonads can disperse anywhere. When bacterial ice nucleation occurs around plant tissue, ice crystals form within the tissue and damages the plant. To examine microbial composition, two Oklahoma lakes: the urban lake- Lake Hefner and the rural lake-Crowder Lake, were examined. In each lake, water samples were collected from three specific locations in summer and winter months. Samples were inoculated on 10% tryptic soy agar to determine total bacterial load and King's B agar supplemented with borie acid, cycloheximide, and cephalexin to isolate Pseudomonads. At Lake Hefner, the eastern shore had significantly larger bacterial concentration in both seasons compared to other locations. At Crowder Lake, the center of the lake had significantly lower bacterial concentration in both seasons compared to other locations. Seasonally, there was lower bacterial concentration in the winter at Lake Hefner compared to the summer, while Crowder Lake had similar bacterial concentration year-round. Lake Hefner had higher bacterial concentration compared to Crowder Lake. Pseudomonads isolated from both seasons and lakes were tested for ice nucleation activity by inoculating in super cooled water and detecting ice formation. Twenty-two ice nucleators were detected at Crowder Lake and three ice nucleators were detected at North to Weatherford 2 miles water molecules. Through evaporation and rainfall, Figure 2. Sampling sites Crowder Lake are marl- stars (Yellow = Near Do- Center of Lake, Green= Shore). • Pseudomonad conc did not differ betwe sites; however, total load was significant the center of compared to the doc (Fig. 3). Lake Hefner in the summer, and none were detected in the winter. The seasonal differences in the concentration of ice nucleating bacteria were surprising because other studies have shown that cooler environments select for increased ice nucleating bacteria. Spatial V Ice Nucleating Pseudomonads Herintal yrimpee yrngan deud ·pringe un erosos om wid and cutivaed pants Lake snow waters Hefner - Figure 4. Sampling sit Lake Hefner are marl stars (Orange = South Shore, Purple = Near Blue = Eastern Shore %3D %3D Ennhment and diversficaon -ayoystem Figure 1. Movement of Pseudomonas syringae throughout the water cycle (Morris et al., 2008). Lake Hefner Pseudomonads are bacteria that inhabit soils and are pathogens of plants and animals. Ice nucleating Pseudomonads (hereafter referred to as Ice+) are bacteria that can freeze supercooled water in their vicinity at a higher temperature than freezing would occur in their absence. Icet strains have been detected in all portions of the water cycle and likely spread via precipitation (Fig. 1) (Morris et al., 2008). Due to the enrichment of Ice+ strains in clouds, ice nucleation activity is hypothesized to allow Pseudomonads to move via clouds in association with aerosolized water. This can be problematic because Ice+ strains can be transmitted to agricultural plants through the water cycle where they cause disease (Morris et al., 2008), Furthermore, Ice+ strains can cause frost injury via ice nucleation of water in plant tissues (Lindow et al., 1982). Ice+ bacteria have been detected on multiple continents, in all forms of water, and in all seasons. (A) 8000 b 6000 4000 2000 Bacterial Loac Figure 6. Seasonal b- Hefner (A) and Crowe Fig. 3. Values indicat value <0.05). Encmant nd gcuttural CFU/ml E
Spatial Variations in Crowder Lake
Novel Ice+ Strains Detected
2000
North to
Weatherford
a
Table 1. Number of Ice+ strains detected in Lake
Hefner and Crowder Lake in winter and summer.
2 miles
1000
1 mile
Summer Ice+Winter Ice+
-1000 Bacterial Load Pseudomonas
Lake Hefner
Crowder Lake
P
22
O Near Dock
IOpposite Shore
Figure 3. Average bacterial
Pseudomonad concentrations at each
Crowder Lake location. Samples were
ICenter of Lake
3
Figure 2. Sampling sites at
Crowder Lake are marked by
stars (Yellow = Near Dock, Red =
Center of Lake, Green Opposite evaluated by determining CFU/ml on
Shore).
Ice+ strains were detected by inoculating isolates in supercooled
water and observing ice crystallization.
The identity of Ice+ strains was determined by 16s FRNA sequencing
(Table 2).
and
10% tryptic soy agar (bacterial load)
Table 2. Identification, previously described Icet status, and known
• Pseudomonad concentrations and King's B agar supplemented with
did not differ between sample
sites; however, total bacterial cephalexin
load was significantly lower in indicated by different letters differ
the
habitats of cultured Icet strains.
boric
acid,
cycloheximide,
and
(Pseudomonas). Values
ID via 16s FRNA Sequencing
Ice+ Status
Habitats
Aeromonadaceae bacterium NRII13
Pseudomonas syringae pv. syringae strain
PS-13
No
Animal/Human/Soil
lake significantly (single factor ANOVA, p-
value <0.05).
center of the
Yes
Animal/Plant Pathogen
compared to the dock or shore
(Fig. 3).
Acinetobacter johnsonii strain P212
Pseudomonas syringae pv. syringae strain
PS-13
Pseudomonas moraviensis strain
HFBPROI
No
Soil
Yes
Animal/Plant Pathogen
Animal/Plant Pathogen
Human wounds
Animal/Plant/Human
Animal/Plant/Human
Yes
Spatial Variations in Lake Hefner
Meromonas veronii B565
Pantoca dispersa strain MLTBY
Pantoea dispersa strain UBI
No
No
10000
No
No
Soil
Acinetobacter calcoaceticus strain HK24
Pseudomonas syringae pv. theae strain
KIWI-ZZ
b b
E 5000
Animal/Plant Pathogen
Plant/Human
Yes
Mixta gaviniae strain DSM 2275
Pseudomonas syringae pv. theae strain
KIWI-ZZ
No
Bacterial Load Pseudomonas
Animal/Plant Pathogen
Yes
-5000
I Southern Shore I Near Dock
I Eastern Shore
Discussion
Lake
Hefner
Figure 5. Average bacterial and
Pseudomonad concentrations in Lake
Figure 4. Sampling sites at
Lake Hefner are marked by
stars (Orange = Southern
Shore, Purple = Near Dock,
Blue = Eastern Shore).
Spatial variations were detected in Crowder Lake (Fig. 3) and
Lake Hefner (Fig. 5).
• Seasonal analysis demonstrates that there is lower overall bacterial
concentration in the winter season for Lake Hefner but not
Crowder Lake (Fig. 6). Neither site differed in the concentration of
Pseudomonads between seasons.
Hefner. Samples were processed and
statistics performed as described in Fig.
3.
• There were no detectable Ice+ strains in these lakes in the winter
months (Table 1). This was a surprise, as previous studies in
multiple locations showed evidence of higher Ice+ activity in
winter months compared to summer months.
• Seven of the twelve Ice+ strains identified do not have previously
published Icet activity, suggesting that their role in ice nucleation
has not been previously characterized (Table 2).
• Four Ice+ isolates identified were plant pathogen P. syringae (Table
2).
Lake Hefner Exhibits Seasonal Variations
(B) 2000
Summer Average
(A) 8000
a
b.
1500
Winter Average
6000
1000
4000
500
2000
Bacterial LoadPseudomonas
Bacterial Load Pseudomonas
References
Figure 6. Seasonal bacterial and Pseudomonad concentrations at Lake
Hefner (A) and Crowder Lake (B). Samples were processed as described in
Fig. 3. Values indicated by different letters differ significantly (T-test, p-
value <0.05).
Lindow, S. E, Arny, D. C, & Upper, C.D. (1982). Bacterial ice nucleation: A factor la frost injury te plants
Plant Physiolegy, 704), 1084-1089, https:/idol.org/10.1104'pp.70.4.184
Morrik, C. E, Sands, D. C. Vinatzer, B. A Glaus, C. Guibaud, C, Buffere, A. Yan, S. Dominguer, H,&
Thompson, H M. (2008). The life history of the plant pathogen pendemenar yringae is laked te the water
cycle. The SME Jeurnal, 2), 321-334. httpst/dol.org/0.1038/smej.2007.13
CFU/ml
CFU/ml
CFU/ml
CFU/ml
Transcribed Image Text:Spatial Variations in Crowder Lake Novel Ice+ Strains Detected 2000 North to Weatherford a Table 1. Number of Ice+ strains detected in Lake Hefner and Crowder Lake in winter and summer. 2 miles 1000 1 mile Summer Ice+Winter Ice+ -1000 Bacterial Load Pseudomonas Lake Hefner Crowder Lake P 22 O Near Dock IOpposite Shore Figure 3. Average bacterial Pseudomonad concentrations at each Crowder Lake location. Samples were ICenter of Lake 3 Figure 2. Sampling sites at Crowder Lake are marked by stars (Yellow = Near Dock, Red = Center of Lake, Green Opposite evaluated by determining CFU/ml on Shore). Ice+ strains were detected by inoculating isolates in supercooled water and observing ice crystallization. The identity of Ice+ strains was determined by 16s FRNA sequencing (Table 2). and 10% tryptic soy agar (bacterial load) Table 2. Identification, previously described Icet status, and known • Pseudomonad concentrations and King's B agar supplemented with did not differ between sample sites; however, total bacterial cephalexin load was significantly lower in indicated by different letters differ the habitats of cultured Icet strains. boric acid, cycloheximide, and (Pseudomonas). Values ID via 16s FRNA Sequencing Ice+ Status Habitats Aeromonadaceae bacterium NRII13 Pseudomonas syringae pv. syringae strain PS-13 No Animal/Human/Soil lake significantly (single factor ANOVA, p- value <0.05). center of the Yes Animal/Plant Pathogen compared to the dock or shore (Fig. 3). Acinetobacter johnsonii strain P212 Pseudomonas syringae pv. syringae strain PS-13 Pseudomonas moraviensis strain HFBPROI No Soil Yes Animal/Plant Pathogen Animal/Plant Pathogen Human wounds Animal/Plant/Human Animal/Plant/Human Yes Spatial Variations in Lake Hefner Meromonas veronii B565 Pantoca dispersa strain MLTBY Pantoea dispersa strain UBI No No 10000 No No Soil Acinetobacter calcoaceticus strain HK24 Pseudomonas syringae pv. theae strain KIWI-ZZ b b E 5000 Animal/Plant Pathogen Plant/Human Yes Mixta gaviniae strain DSM 2275 Pseudomonas syringae pv. theae strain KIWI-ZZ No Bacterial Load Pseudomonas Animal/Plant Pathogen Yes -5000 I Southern Shore I Near Dock I Eastern Shore Discussion Lake Hefner Figure 5. Average bacterial and Pseudomonad concentrations in Lake Figure 4. Sampling sites at Lake Hefner are marked by stars (Orange = Southern Shore, Purple = Near Dock, Blue = Eastern Shore). Spatial variations were detected in Crowder Lake (Fig. 3) and Lake Hefner (Fig. 5). • Seasonal analysis demonstrates that there is lower overall bacterial concentration in the winter season for Lake Hefner but not Crowder Lake (Fig. 6). Neither site differed in the concentration of Pseudomonads between seasons. Hefner. Samples were processed and statistics performed as described in Fig. 3. • There were no detectable Ice+ strains in these lakes in the winter months (Table 1). This was a surprise, as previous studies in multiple locations showed evidence of higher Ice+ activity in winter months compared to summer months. • Seven of the twelve Ice+ strains identified do not have previously published Icet activity, suggesting that their role in ice nucleation has not been previously characterized (Table 2). • Four Ice+ isolates identified were plant pathogen P. syringae (Table 2). Lake Hefner Exhibits Seasonal Variations (B) 2000 Summer Average (A) 8000 a b. 1500 Winter Average 6000 1000 4000 500 2000 Bacterial LoadPseudomonas Bacterial Load Pseudomonas References Figure 6. Seasonal bacterial and Pseudomonad concentrations at Lake Hefner (A) and Crowder Lake (B). Samples were processed as described in Fig. 3. Values indicated by different letters differ significantly (T-test, p- value <0.05). Lindow, S. E, Arny, D. C, & Upper, C.D. (1982). Bacterial ice nucleation: A factor la frost injury te plants Plant Physiolegy, 704), 1084-1089, https:/idol.org/10.1104'pp.70.4.184 Morrik, C. E, Sands, D. C. Vinatzer, B. A Glaus, C. Guibaud, C, Buffere, A. Yan, S. Dominguer, H,& Thompson, H M. (2008). The life history of the plant pathogen pendemenar yringae is laked te the water cycle. The SME Jeurnal, 2), 321-334. httpst/dol.org/0.1038/smej.2007.13 CFU/ml CFU/ml CFU/ml CFU/ml
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