JOURNAL

OF INVERTEBRATE

PATHOLOGY

(1990)

s&417-427

Effect of Water Potential, Temperature, and Clay-Coating on Survival

of Beauveria bassiana Conidia in a Loam and Peat Soil
JOHN P. STUDDERT'ANDHARRY
Departments of Entomology and Nematology,

K. KAYA

University of California,

Davis, California

95616

AND
JOHN M. DUNIWAY
Department of Plant Pathology,

University of California,

Davis, California

95616

Received May 10, 1989; accepted September 11, 1989

When Beauveria bassiana conidia were mixed in nonsterile Yolo tine sandy loam (YFSL) or
Staten peaty muck (peat) soil at water potentials ranging from 0.0 bars (saturation) to - 1500 bars,
conidia half-lives were longest at - 15 bars and decreased as the water potential approached either
0.0 or -200 bars. Conidia half-lives increased as the water potential decreased from -200 to
- 1500 bars. Conidia half-lives were longest at a soil temperature of 10C and decreased both at 2C
and as the temperature approached 50C where no conidia were recovered after 2 weeks. The
longest mean half-life value was 44.4 weeks for conidia in YFSL at - 10 bars and 10C; the shortest
half-life value was 0.3 weeks in peat soil at 0 bars and 28C. Clay-coating lengthened conidia
survival in all treatments in a factorial experiment involving the two soil types and several combinations of soil temperature and water potential. When two strains of B. bassiana were compared,
colony counts of strainABG-6178 always decreased relative to an initial baseline count taken soon
after mixing conidia and soil; colony counts of strain IL-l 16 routinely increased after the baseline
count was taken before decreasing later. Conidia survival was often significantly longer in the low
organic YFSL than in the high organic peat. The results suggest that conidia survival is influenced
by the direct effect of physical factors and soil microbial populations. Since B. bassiana conidia
survival is highly variable, its potential as a microbial insecticide is much greater in some soil
environments than in others. 6 1990Academic press, Inc.
KEY WORDS: Beauveria bassiana; conidia survival; conidia half-life; soil water potential; soil
temperature; clay-coating; microbial insecticide.

INTRODUCTION

The entomopathogenic
fungus, Beauveria bassiana, is a potentially important
microbial insecticide in the soil environment achieving partial control of the Colorado potato beetle, Leptinotarsa decemlineata (Watt and LeBrun, 1984), the pecan
weevil, Curculio caryae (Gottwald and
Tedders, 1983), and the curculionid, Sitona
lineatus (Mtiller-Kogler
and Stein, 1970).
To be effective, B. bassiana conidia must
remain viable and infect insect hosts in the
soil under a wide range of physical and biological conditions.
Studies have deter Present address: Cooperative Extension, University of California, 142A Garden Highway, Yuba City,
California 95991.

mined the effect of temperature, relative

humidity, and light on the survival of entomopathogenic Deuteromycetes conidia in a
nonsoil setting (Kawakami,
1960; Steinhaus, 1960; Miiller-Kogler,
1964; Clerk and
Madelin, 1965; Walstad et al., 1970; Daoust
and Roberts, 1983). Soil-based studies with
B. bassiana have been limited to the longevity of conidia (Lingg and Donaldson,
1981; Fargues et al., 1983; Fargues and
Robert, 1985; Miiller-Kogler
and Zimmermann, 1986). However, Lingg and Donaldson (1981) determined the effect of physical
factors (soil moisture content, relative humidity, temperature, and pH) on conidia
survival in the soil using moisture content
as a percentage of the saturation capacity of
the soil. Lingg and Donaldson (1981) and
417
0022-2011&O $1.50
Copyright 0 1990 by Academic Press, Inc.
All rights of reproduction in any form reserved.

418

STUDDERT,

KAYA,

Fargues et al. (1983) also demonstrated the

importance of soil-biotic factors on conidia
survival.
Soil moisture has important effects on
microbial activity. Unfortunately,
much of
the early research evaluating soil moisture
effects on microorganisms cannot be interpreted adequately because soil moisture
content was usually expressed in noncomparable terms (e.g., percentage of waterholding capacity or percentage of soil dry
weight). Furthermore, these soil moisture
measurements are not related to the availability of water to soil microorganisms
(Griffin, 1963). Water potential is a more
meaningful measure biologically because
two soils with the same water potential
make water equally available to soil microorganisms even if their water, measured as
a percentage of soil dry weight, differ (Griffin, 1963; Sommers et al., 1981). Water potential is defined as the chemical potential
of water per unit volume and has the dimensions of pressure. The units are usually bars
or megapascals (Duniway, 1983; Griffin,
1963, 1981). In recent years water potential
has been used extensively to define soil water status in research conducted by plant
pathologists and soil microbiologists (e.g.,
Cook and Duniway, 1981; Duniway, 1976,
1983; Duniway and Gordon, 1986; Griffin,
1981; Sommers et al., 1981).
The objective of this study was to determine relationships between soil water potential and temperature and the survival of
B. bassiuna conidia in two nonsterile soils,
a low organic sandy loam and a high organic
peat. This was done for both clay-coated
and noncoated conidia and for two different
strains of B. bassiana.
MATERIALS

AND METHODS

B. bassiana strains. Two strains of B.

bassiuna conidia, ABG-6178 and IL-l 16,
were obtained from Abbott Laboratories,
North Chicago, Illinois. The conidia were
stored in the dark at 10C and 0% relative
humidity (RH). Unless otherwise stated,

AND

DUNIWAY

only ABG-6178 was used because it was

being developed as a microbial insecticide.
Soil types. Two soil types, Yolo fine
sandy loam (YFSL) (cl% organic matter)
and Staten peaty muck (peat) (62% organic
matter), were used. The YFSL came from
the Botany Department (stored outdoors
and exposed to ambient conditions) at the
University of California at Davis; the peat
came from an abandoned field near Terminous, California. The percentage sand, silt,
andclaywas64.5,23.4,
11.6and12.1,
11.4,
14.4 in YFSL and peat, respectively. Water
release curves showing the relationship between soil matric potential and percentage
water content were determined for both
soils (Fig. 1). Each soil was sieved (2-mm
mesh) to remove stones and large particles
and mixed in a cement mixer to achieve
uniformity throughout the soil mass. Both
soils were stored air-dried at 24C until experimental
matric and water potentials
were established. The soils were not sterilized.
Water and matric potential determination. Saturated soil has a water potential at
nearly 0 bars. As soil becomes drier, the
water potential becomes increasingly negative. Soil water potential has two major
components, matric potential and solute
potential. The capillary and absorption
forces associated with the soil matrix constitute the matric potential. The matric potential is essentially equal to the water potential as long as a soil is low in salt. The
solute potential consists of the osmotic
forces caused by salt in the soil solution. In
our experiments, matric potentials were established between 0 and - 15 bars; water
potentials were established for the -2OOand - 1500-bar soils.
Matric potentials of 0, -0.1, and -0.3
bars were established using Buchner funnels of 9-cm diameter with fritted glass
plates of fine porosity (KIMBLE
284009OF) as tension plates (Duniway, 1976). The
desired matric potential was obtained by
adjusting the height of a water column running between the surface of a water reser-

SURVIVAL

OF Beauveria

IN

419

SOIL

pziq

160
ii?
xi
E

120-

8
ti

EOw

"

I
0

-3

I
- 6

Soil Math

I
- 9

I
- 12

0
I
- 15

I
- 18

Potential (bars)

FIG. 1. Soil water content (percentage calculated as g water/100 g dry soil) of Yolo fine sandy loam
(YFSL) and Staten peaty muck (peat) plotted as functions of decreasing soil matric potential.

voir and the tension plate. Matric potential

was established in the Buchner funnels in
less than 24 hr. The -0.3-bar soil was removed from the Buchner funnels after matric potential was established and equilibrated for 12 weeks using the procedures
employed for the -2, - 10, and -U-bar
soils (see below). After this time period B.
bassiana
conidia
were mixed in the
-0.3-bar soil, and within l-3 hr, and the
first inoculated soil samples were taken for
the baseline dilution series for the conidia
survival experiments.
It was not possible to follow the same
procedure with 0 and -0. l-bar soils because they were too wet to mix with conidia. For these matric potentials, the conidia
were initially mixed in - IO-bar soil which
was placed in Buchner funnels where 0 and
-0.1 bar matric potentials were established. Immediately thereafter the first soil
samples were taken from the Buchner funnels for a baseline dilution series. Thus, the
0 and - 0. l-bar soils did not undergo a long
equilibration period before the mixing of
soil with conidia as did the soils at the other
matric and water potentials.
A pressure plate apparatus (Soil Moisture Equipment Co., Santa Barbara, Cali-

forma) was used for the determination of

soil water content at -2, - 10, and - 15
bars (Grifftn, 1963). These matric potentials
were subsequently established by first mixing air-dried soil (2000 g) with appropriate
amounts of water to create moist soil. This
moist soil, after equilibrating for 3 weeks,
was mixed with appropriate amounts of airdried soil to adjust the net moisture content
of the soil by weight to give the final matric
potential. For this work, water was added
to the soil with a hand-held mister. All mixing of soil and water or moist soil and airdried soil was done by hand inside polyethylene sacks to prevent moisture loss. The
soil was then allowed to equilibrate for 12
weeks inside polyethylene sacks wrapped
in cotton (to reduce condensation on the
inside of the soil sacks), covered with damp
paper towels (wetted weekly), and kept in
the dark in constant temperature chambers.
Soil samples taken monthly showed water
losses were no more than 5% of the original
soil moisture content in any soil sack over
the course of the experiments. The long
equilibration period was necessary to create a uniform water potential throughout
the soil mass. It may also have allowed distinct microbial populations, associated with

420

STUDDERT,

KAYA,

each soil environment, a long time period to

develop prior to adding conidia to the soil.
Soil water potentials of - 200 and - 1500
bars were established by placing 200-g
quantities of air-dried soil to a depth of 2 cm
in open plastic containers inside desiccators over saturated KC1 and MgCl, solutions, respectively.
These soils were allowed to equilibrate for up to 20 weeks before the soil water content became stable.
The KC1 and MgCl, solutions produce
RH values of 86 and 33%, respectively
(Greenspan,
1977). Because RH established over saturated salt solutions varies
with temperature, the - 200- and - 1500bar values were approximate. For soils between -0.3 and - 1500 bars, conidia and
soil were mixed after the equilibration period .
Soil inoculation
with conidia and soil
storage. All soils were inoculated with dry
conidia by hand, using a tongue depressor
to mix the soil and conidia, inside polyethylene sacks to prevent moisture loss. From
previous experience, 20 min of mixing were
sufficient to evenly distribute conidia in
1000 g of soil.
For the duration of the survival experiments the 0- and - 0. l-bar-soils were left in
the Buchner funnels, -0.3-,
-2-, and
- U-bar soils were left in the sacks where
soil and conidia were mixed, and the - 200and - 1500-bar soils remained in the desiccators where water potential was established. The soil sacks, desiccators, and
Buchner funnels were maintained inside
constant temperature chambers during both
the equilibration period and the survival experiments (at either 16 or 28C). For each
treatment, soil was stored in four containers: either four Buchner funnels, each holding ca. 350 g of soil; or four polyethylene
sacks, each holding ca. 1000 g of soil; or
four open containers inside desiccators,
each holding ca. 200 g of soil.
Dilution series and colony counts. To
test the survival of B. bassiana conidia in
the soil, factorial experiments using different soil water content and temperature val-

AND

DUNIWAY

ues were conducted. Five-gram soil samples were taken, one from each of the four
soil storage containers constituting a treatment each time a dilution series was conducted. Each soil sample was stirred in 250
ml of 0.1% agar water used to keep the soil
in suspension. After 3 hr, 5 ml of the original suspension was pipetted into 200 ml of
0.1% agar water; 2 hr later, 1.25 ml was
taken from the second suspension and pipetted on each of three Petri plates containing a selective medium made with the fungicide dodine (Chase et al., 1986). The Petri
plates were stored in the dark at 24C for
6-7 days prior to counting all B. bassiana
colonies on each plate.
Except for the 0- and -0. l-bar matric
potentials, the baseline dilution series were
started within 1-3 hr of mixing conidia and
soil. For the 0- and -O.l-bar
soils, the
baseline dilution series were begun 24 hr
after mixing soil and conidia, to allow matric potentials to establish in the Buchner
funnels. Later dilution series and colony
counts were conducted 2 and 5 weeks after
the baseline series and at 5-week intervals
thereafter until no conidia could be recovered from a treatment or until 50 weeks had
passed. In the comparison of two fungal
strains, colony counts were also made 1
week after the baseline series. Sufftcient
numbers of conidia were mixed with the
soil so that the baseline colony counts
ranged from 300 to 400 colonies per plate.
Clay-coating of conidia. B. bassiana conidia were washed in distilled water, allowed to dry, and coated by mixing conidia
with a bentonite clay (1:3 by weight) provided by Mycogen Corp., San Diego, California. This mixture was spread out flat on
a plastic surface, sprayed lightly with sterile distilled water from a hand-held mister,
and allowed to dry for 72 hr in the dark. The
dry mixture was used in experiments where
the survival of clay-coated and noncoated
conidia were compared.
Calculations
of conidia half-lives and
statistical analysis. Conidia half-lives were
calculated using the procedures employed

SURVIVAL

OF Beauveria

by Lingg and Donaldson (1981). A half-life

was calculated for each curve generated by
the decrease in B. bassiuna (strain ABG6178) colony counts over time. Four such
curves were generated per treatment, one
curve from the soil samples taken from
each of the four soil containers that constituted a treatment. These half-life values
were subjected to a logarithmic transformation prior to an analysis of variance and
Duncans multiple-range test. The 5% level
of significance was employed for all data.
Terminology.
During the discussion of
experimental procedures, care was taken to
maintain the distinction between matric and
water potential. Since our soils were low in
salt (peat has < 1.0 mmhos/cm and YFSL
~0.2 mmhos/cm), the difference between
water potential and matric potential was
small so the term water potential is used in
the remainder of the paper.
RESULTS

B. bassiana strain and conidia survival.

Two strains of B. bassiana responded very
differently in a dilution series experiment
(Figs. 2A and 2B). The ABG-6178 colony
counts decreased over time in all trkatments. With the IL-116 strain an initial increase in colony counts, relative to the
baseline count, occurred for all combinations of soil type and water potential. This
increase was apparent at 1 week following
the baseline count. At 2 weeks counts were
more than twice the baseline count. From
that point on IL-l 16, colony counts decreased at a more rapid rate than the decrease shown by ABG-6178 counts so that
recovery of the two strains ceased at about
the same time. Since populations of ABG6178 always decreased in a manner approximating first-order
kinetics, survival of
conidia of this strain was expressed in halflives to compare treatments in the experiments discussed below.
Soil water potential

and conidia survival.

When B. bassiana conidia were subjected

to water potentials ranging from 0 to - 15
bars, half-lives were longest at - 15 bars

IN

SOIL

421

and decreased as the soil became wetter,

i.e., as water potential increased to 0 bars
(Table 1). At saturation (0 bars) and -0.1
bars, conidia were recovered in all treatments at 2 weeks, but no recovery occurred
at 5 weeks. All conidia half-lives at these
high water potentials
were less than 1
week. As water potential decreased from
-0.1 to - 15 bars, conidia half-lives increased more rapidly at 16C than at 28C
(for YFSL and peat) and more rapidly for
YFSL than for peat (at 16 and 28C). As a
result, conidia half-lives reached their highest values of 36.3 and 15.2 weeks at 16C
and - 15 bars in YFSL and peat, respectively. When soil water potential was decreased from - 15 to - 200 bars, there were
significant decreases in conidia half-lives in
all treatments. At -200 bars conidia halflives ranged from 1.1 to 3.5 weeks. However, as water potential decreased further
to - 1500 bars, conidia half-lives increased
again and were significantly higher than
were the corresponding
values at -200
bars. At - 1500 bars, conidia half-lives
ranged from 5.3 to 7.9 weeks. At all water
potentials, conidia survival was longer in
YFSL than in peat (at both temperatures)
and longer at 16C than in 28C (for both
soil types). These differences were usually
significant in the middle range of water potentials between -0.3 and - 15 bars and
usually not significant in the wetter and
drier soils.
Soil temperature and conidia survival. B.
bassiuna conidia (strain ABG-6178), mixed

in soil and subjected to a range of temperatures from 2 to 50C survived longest at

10C (Tab!e 2). Conidia half-lives then decreased steadily as temperatures increased
from 10 to 50 until no recovery occurred
at 2 weeks at 50C. Conidia half-lives were
significantly
less for all 2C treatments
when compared to the corresponding 10C
treatment. At 2, lo, and 20C conidia survival was significantly longer in YFSL than
in peat (at -0.3 and - 10 bars) and significantly longer at - 10 bars than at - 0.3 bars
(for YFSL and peat). At 30, 40, and 50 no

422

STUDDERT,

KAYA,

AND DUNIWAY

I YFSL

28 C

,e

-1OBars

-0.3 Ban

IL-116
IL-1 16

-10 Bars

ABG-6176

-0.3 Bars

ABG-6176

100

50

t
.5
z
m
z

;
iti
t
P

0
0

10

15

25

20

250

B
PEAT

200

-o-

150

28 C
-1OBars

IL-116

--)-

-0.3 Bars

IL-116

---(>-

-1OBars

ABG-6176

--f-

-0.3 Bars

ABG-6178

100

50

0
0

10

15

20

25

Time (Weeks)
FIG. 2. Beauveriu bassiana colony counts made over time and expressed as a percentage of the
baseline colony counts taken at Week 0. The graphs show the results of dilution series made periodically after mixing strain ABG-6178 or IL-116 conidia in (A) Yolo tine sandy loam (YFSL) and (B)
Staten peaty muck (peat). The soils were maintained at 28C with a water potential of -0.3 or - 10
bars. Each percentage is based on four dilution series, one of each of four soil containers making up
a treatment, and three colony counts for each dilution series.

SURVIVAL

OF Beauveria
TABLE

SURVIVAL

(HALF-LIVES

IN WEEKS)

OF Beauveria

423

IN SOIL

bassiana

CONIDIA

IN NONSTERILE

SOIL,

EITHER

YOLO

FINE SANDY LOAM (YFSL) OR STATEN PEATY MUCK (PEAT), AT DIFFERENT WATER POTENTIALS
AND TEMPERATURES

Temperature
(Cl

Soil
YFSL

16
28
16
28

Peat

Water potential (bars)

0.0
0.6
0.4
0.5
0.3

-0.1

Aa
Aa
Aa
Aa

0.9
0.5
0.1
0.4

-2.0

-0.3

Aa
Aa
Aa
Aa

15.0
4.9
5.1
2.1

Ad
Bb
Bb
Cbc

26.7
5.1
13.8
2.1

Ae
Cb
Bc
Dbc

- 15.0

-200.0

- 1500.0

36.3 Af
5.4 Cb
15.2 Bc
3.1 Ccd

3.5 Ab
1.2 Ba
1.3 Ba
1.1 Bab

7.9
6.3
6.9
5.3

AC
Ab
Ab
Ad

e Mean of four half-life values in weeks. Each half-life value was calculated from the conidia survival curve
generated from B. bassiana colony counts from a single soil container. There were four containers per treatment.
Means followed by different uppercase letters in a column and by different lowercase letters in a row are
significantly different (P < 0.05) according to Duncans multiple-range test.

significant differences in conidia half-lives,

related to water potential or soil type, were
observed.
Clay-coating and conidia survival. In all
cases, regardless of soil type, water potential, or temperature,
conidia half-lives
(strain ABG-6178) were significantly longer
in treatments
using clay-coated conidia
than in the corresponding treatments with
noneoated conidia (Table 3). For both claycoated and noncoated conidia at 10C survival was significantly longer in YFSL than
in peat, regardless of water potential, and
significantly
longer at - 15 bars than at
- 0.3 bars, regardless of soil type. At 30C
the spread between the shortest mean halflife value (2.0 weeks for noncoated conidia

in peat at -0.3 bars) and the longest mean

half-life value (12.2 weeks for clay-coated
conidia in YFSL at - 15 bars) was much
less than the corresponding spread at 10C
(11.8 to 64.4 weeks). As a result of the relatively low half-life values at 30C most of
the significant differences related to soil
type and water potential at 10C did not occur at 30C. Also for clay-coated and noncoated conidia, half-lives were significantly
longer at 10C than in the corresponding
30C treatment, regardless of soil type or
water potential.
DISCUSSION
B. bassiana strain and conidia survival.
We cannot adequately explain why strain

TABLE

CONIDIA
IN NONSTERILE
SOIL, EITHER YOLO
FINE SANDY LOAM (YFSL) OR STATEN PEATY MUCK (PEAT), AT DIFFERENT TEMPERATURES AND

SURVIVAL

(HALF-LIVES

IN WEEKS~)

OF Beauveria

bassiana

WATER

Soil
YFSL
Peat

Water
potential
(bars)

- 10.0
-0.3
- 10.0
-0.3

21.6
12.2
16.9
8.3

POTENTIALS

Temperature (C)
10
Ad
cc
Bd
DC

44.4
21.8
23.4
13.9

Ae
Bd
Be
Cd

20
20.8
10.9
10.7
6.9

AC
Bc
Bc
Cc

30
4.3
3.9
3.3
3.0

Ab
Ab
Ab
Ab

40
1.8
1.6
1.1
0.8

Aa
Aa
Aa
Aa

50
NRb
NR
NR
NR

Mean of four half-life values in weeks. Each half-life value was calculated from the conidia survival curve
generated from B. bassiana colony counts from a single soil container. There were four containers per treatment.
Means followed by diierent uppercase letters in a column and by different lowercase letters in a row are
significantly different (P < 0.05) according to Duncans multiple-range test.
b NR, no recovery at 2 weeks after conidia were mixed in soil.

424

STUDDERT,

KAYA,

AND DUNIWAY

TABLE 3
SURVIVAL (HALF-LIVES IN WEEKSO) OF CLAY-COATED (CC) AND NONCOATED (NC) Beauveria bassiana
CONIDIA IN NONSTERILE SOIL, EITHER YOLO FINE SANDY LOAM (YFSL) OR STATEN PEATY MUCK (PEAT),
AT DIFFERENT WATER POTENTIALS AND TEMPERATURES
Temperature
cc,
10
30

YFSL
Conidia
cc
NC
cc
NC

-0.3 Bars
31.4
26.1
11.4
4.6

Ab
Bb
Cb
Dab

Peat
- 15.0 Bars
64.4 AC
44.2 Bc
12.2 Cb
6.6 Dbc

-0.3 Bars
20.3
11.8
6.6
2.0

Aa
Ba
Ca
Da

- 15.0 Bars
34.2
26.8
9.7
4.4

Ab
Bb
Cab
Dab

LIMean of four half-life values in weeks. Each half-life value was calculated from the conidia survival curve
generated from the B. bassiana colony counts from a single soil container. There were four containers per
treatment. Means followed by different uppercase letters in a column and by different lowercase letters in a row
are significantly different (P < 0.05) according to Duncans multiple-range test.

IL-116 colony counts increased to about

200% of their baseline values over a 2-week
period before declining. Baseline colony
counts, using ABG-6178 conidia, were usually within 15% of values expected when
extrapolating from the number of conidia
per gram in the storage containers. For the
IL-116 strain, the baseline colony counts
were about 25% of what would be expected
when extrapolating from the storage container counts. Possibly, at the time of the
baseline dilution series, the IL-116 conidia
were clumped in the soil, each clump giving
rise to a single B. bassiuna colony. Later,
less clumping occurred and colony counts
increased. If such artifacts are operating,
care must be taken in interpreting colony
count numbers. Fargues and Robert (1985)
speculated that the survival of Metarhizium
unisopliue
is influenced by microcyclic
conidiation in the soil, but we have no evidence that microcyclic conidiation could
account for the survival pattern of strain
IL- 116. Miiller-Kiigler
and Zimmermann
(1986) also obtained an unexplained increase in B. bussiunu colony counts from
soil dilution series over time.
Soil water potential

and conidiu survival.

In nonsoil experiments, Clerk and Madelin

(1965) and Daoust and Roberts (1983)
showed that the viability of M. unisopliue
conidia first decreased and then later increased as equilibrium RH decreased from
98 to 92% (- 25 to - 110 bars water poten-

tial) to 0% (0 bars). The reason for this phenomenon is unknown. The survival of B.
bussiunu conidia in the soil follows a similar
pattern. In our study, B. bussiunu conidia
half-life values decreased from a high point
at - 15 bars (98.9% RH) to significantly
lower values at - 200 bars (86% RH). However, as the water potential decreased further to - 1500 bars (33% RH), conidia halflives increased again. Since there is little
microbial activity in soils drier than -50
bars (Wilson and Griffin, 1975), the explanation for this phenomenon
is probably
physiological in nature.
Lingg and Donaldson (1981) pointed out
that soil RH often is not drier than 98.9%
( - 15 bars). This is certainly true in agricultural soils where moisture levels are kept
high enough to support crop growth. Since
many plant species reach their permanent
wilting point at ca. - 15 bars, RH values
below 98.9% usually do not occur in these
soils except at the soil-air interface. However, in soil environments where there are
long periods without precipitation,
water
potential can reach values at least as low as
- 100 bars (92.9% RH) (e.g., Cook and
Duniway, 1981). The survival of B. bussiunu conidia will be influenced to the extent
that natural soils in dry areas reach these
low water potentials.
A review by Sommers et al. (1981)
showed that the most extensive microbial
(mostly bacterial) decomposition of organic

SURVIVAL

OF Beauveria

material occurs in soils at water potentials

ranging from -0.1 to -0.3 bars. Extensive
bacterial decomposition can also occur in
wetter soils at or approaching saturation (0
bars). Bacterial movement, the ability to
lyse fungal mycelium, and bacterial respiration become negligible in all soils by the
time water potential decreases to - 15 bars
Wilson and Griffin, 1975; Grifftn, 1981).
Wilson and GrifIin (1975) also observed a
rapid decline in total soil microbial respiration between -3 and -8 bars, mostly due
to reduced bacterial activity. Between -8
and -30 bars total microbial respiration
(mostly fungal respiration) declined slowly
before decreasing rapidly and becoming
negligible at - 50 bars. Thus, it seems likely
that the low conidia half-lives in our study
at 0 and -0.1 bars were at least partly due
to the destruction of B. bassiana conidia by
bacteria. The increase in conidia viability
as water potential decreased from - 0.1 to
- 15 bars may represent decreased destruction of conidia by soil microorganisms as
moisture conditions became drier and less
favorable to their activity. However, because the direct effects of high water potentials on B. bassiana conidia are unknown,
there may also be physiological limitations
on conidia survival in very wet soil.
Fargues et al. (1983) demonstrated that
clay-coating increases B. bassiana blastospore survival by protecting
against
bacterial lysis in soils at 80% of moisture-holding capacity. Unless clay-coating
changes the water potential experienced by
blastospores, this shows that they can survive the direct effect of water potentials
close to saturation for longer time periods
when the effect of bacterial lysis is reduced.
We do not have similar data for B. bassiana
conidia, but clay-coating significantly enhanced B. bassiuna conidia survival in soil
at -0.3 and - 15 bars (Table 3).
Since B. bassiana conidia did not survive
well in our soil at 0 and -0.1 bars, the effectiveness of B. bassiana, as a microbial
insecticide, will likely be reduced in very
wet soils. On the other hand, since conidia

IN

425

SOIL

survival significantly increased at water potentials between -0.3 bars (field capacity
for many soils) and - 15 bars (permanent
wilting point for many plants), it appears
that B. bassiuna conidia survive relatively
well over the range of water potentials occurring most frequently
in agricultural
soils. Also, because clay-coating of B. bassiuna conidia increases half-lives over a
wide range of water potentials, we conclude that conidia survival will often be sufficient to use B. bassiuna as an insecticide
in the soil.
Soil temperature

and conidia

survival.

The importance of temperature to the survival of B. bassiana conidia outside of soil

has been demonstrated in several studies
showing sharp decreases in conidia survival
as temperature increased from ca. 8C to
the thermal death point at 50C (Clerk and
Madelin, 1965; Steinhaus, 1960; Walstad et
al., 1970).
Soil temperature probably also had an
important indirect effect on conidia survival in our experiments. Microbial decomposition of soil organic matter is greatest at
temperatures from ca. 18 to 35C when water potential is between saturation and - 15
bars (Nyhan, 1976; Wildung et al., 1975).
Accordingly, the decrease in B. bassiana
conidia survival associated with increasing
temperature in our experiments, especially
at temperatures within the optimum range
for microbial activity, may have been due
to increased microbial degradation of these
conidia.
To the best of our knowledge, the survival of B. bassiana conidia has not been
tested previously at 2C. Our finding that
conidia half-lives were less at 2C than at
10C is contrary to several nonsoil studies
where conidia survival has been shown to
increase as the temperature
approaches
freezing (Muller-Kogler,
1964). For M. anisopliue, at most temperatures, conidia survival was longest at 0% and 97-98% RH
and shorter at intermediate RH values; but,
at 4C viability was less at 98% than at
some intermediate RH values (Daoust and

426

STUDDERT,

KAYA,

Roberts, 1983).As a result, conidia survival

at 98% RH was shorterat 4C than at higher
temperatures.If the same relationship betweenmoisturelevel and temperatureholds
for B. bassianaconidia, it could explain the
observed decreasein conidia half-lives in
our experiment at 99.9% RH (-0.3 bars)
and 99.3% RH (- 10bars) as the temperature approachedfreezing.
Conidia half-lives were less than 1 month
at temperaturesof 30C or higher for all
combinationsof soil type and water potential. This rapid loss of conidia viability at
higher temperatureswould make it difficult
to use B. bassiana as a microbial insecticide in the soil in warm climates. This problem may be partly overcomeby using claycoated conidia since our results demonstrate the usefulnessof this technique for
prolongingthe life of B. bassiana conidia in
the soil.
Soil type and conidia survival. Because
we measured the moisture levels in our
soils in terms of water potential, it was possible to compareconidia survival in two different soils over a wide range of moisture
and temperature conditions. In all of our
experiments,conidia half-lives were significantly longer in YFSL than in the correspondingpeattreatmentat the middle range
of water potentials (-0.3 to - 15bars)and
at temperaturesup to and including 20C.
At the more extreme water potentials and
at the higher temperatures, these differenceswere no longer significant. The peat
soil had a much higherorganiccontentthan
the YFSL and high organicsoils often have
high populations of bacteria, fungi, and
actinomycetes which show significant antagonisticactivity againstsomefungal plant
pathogens (Weste and Vithanage, 1978;
Malajczuk, 1983), and an entomopathogenie fungus(Lingg and Donaldson, 1981).
Thus, high populations of microorganisms
may account, in part, for the relatively
short conidiahalf-lives observedin our peat
soil.
Our results indicate that B. bassiana conidia will survive better in some soil types

AND

DUNIWAY

than in others. For this reason, it may be

necessary to initially determine conidia
half-lives in soils where long-term survival
is important, so that the proper quantities
of conidia can be added to soil where B.
bassiana is to be used as an insecticide.
ACKNOWLEDGMENTS
We thank Dr. M. Stimmann and Dr. E. Butler for
reviewing initial drafts of the manuscript. The senior
author was supported by grants from the IR-4 program
and scholarships from the California Nurserymens
Association.

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