Beruflich Dokumente
Kultur Dokumente
(1990), 115,
BY T . P. M C G O N I G L E S M . H.
Previously described methods to quantify the proportion of root length colonized by vesicular-arbuscular (VA)
mycorrhizal fungi are reviewed. It is argued that these methods give observer-dependent measures of colonization
which cannot be used to compare, quantitatively, roots examined by different researchers. A modified method is
described here to estimate VA mycorrhizal colonization on an objective scale of measurement, involving inspection
of intersections between the microscope eyepiece crosshair and roots at magnification x 200; it is referred to as
the magnified intersections method. Whether the vertical eyepiece crosshair crosses one or more arbuscules is
noted at each intersection. The estimate of colonization is the proportion of root length containing arbuscules,
called the arbuscular colonization (AC). The magnified intersections method also determines the proportion of
root length containing vesicles, the vesicular colonization (VC), and the proportion of root length containing
hyphae, the hyphal colonization (HC). However, VC and HC should be interpreted with caution because vesicles
and hyphae, unlike arbuscules, can be produced in roots by non-mycorrhizal fungi.
Key words: Magnified intersections, percent infection, fractional infection, method, arbuscules.
INTRODUCTION
error of between 2 and 5 % colonization for subjective visual estimation. However, no investigation
has yet been undertaken to assess the accuracy of this
method. In addition, this method has not yet been
considered in terms of the relationship between
perceived magnitude (^) and stimulus magnitude
{(j)), known as Stevens' power law (Teghtsoonian &
Teghtsoonian, 1971):
^ = hij/,
(1)
496
497
Root
Direction
of
travel
across
slide
Horizontal
crosshair
Perimeter
of field
of view
Vertical
crosshair
ib)
Position to which
vertical crosshair
is moved
A N P 115
498
Table 1. A summary of the variables in the investigation into subjective visual estimation using test diagrams {a)
and {b)
Variable
Score for {a)
Score for {b)
Score for (6)
score for {a)
Score for {b)
-r-score for (a)
True
value
Subject
mean
Standard
deviation
Coefficient of
variation (%)
0-45
0-63
0-18
0-49
0-66
0-17
0-11
0-11
0-09
22
17
53
1-40
1-37
0-23
17
0-8
07
0-6
o
CO
0-5
0-4
0-3
0-2
04
0-6
Score for (b)
08
10
RESULTS
The mean scores for diagrams {a) and {b) were close Two examples of the data and of the calculation of
to their true values, although there was considerable AC, VC, and HC are given in Table 2. For the 30
variation across the subjects (Table 1). Despite this maize field samples AC ranged from 010 to 0-70, but
499
Table 2. Examples of data obtained using the magnified intersections method. The subsamples shown are for
separate maize root samples
Number of intersections
Root
subsample
1
Microscope
slide
A
B
C
D
Sum
A
B
C
D
Sum
Negative"
59
54
46
43
Arbuscules*
Vesicles"
Hyphae
only"*
Total'
73
0
0
0
6
6
67
56
4
20
6
52
248
62
44
51
43
200
202
33
25
5
25
23
13
21
23
102
16
14
66
1
0
1
1
13
30
Calculation of arbuscular colonization (AC), vesicular colonization (VC), and hyphal colonization (HC) proceeds as
follows. T h e respective values for subsamples 1 and 2 for AC are 25/248 = 0 1 0 and 66/200 = 0-33; for VC are
1/248 = 0-004 and 2/200 = 0-010; and for H C are ( 2 4 8 - 2 0 2 ) / 2 4 8 = 0-19 and ( 2 0 0 - 1 0 2 ) / 2 0 0 = 0-49.
"Negative: crosshair did not cut through any hyphae nor arbuscules nor vesicles.
"Arbuscules: crosshair cut through at least one arbuscule.
''Vesicles: crosshair cut through at least one vesicle.
''Hyphae only: crosshair cut through at least one hypha, but no arbuscules nor vesicles.
''Total: total number of intersections examined. This must be recorded separately and not calculated as a sum of
categories a, b, c and d, because when examining a given intersection a score of 1 can sometimes be added to both
categories b and c
Pot
AC
VC
HC
10
11
12
0-31 (0-06)
0-27 (0-05)
0-38 (0-03)
0-31 (0-07)
0-23 (0-08)
0-34 (0-05)
0-66 (0-04)
0-66 (0-10)
0-71 (0-07)
500
Table 4. Means and standard deviations fmr arbuscular colonization {AC) and hyphal colonization {HC) in the
study to assess the variability within a subsample, between subsamples, and between observers. A separate mai
root sample was used for each of the four cases {n = sample size)
AC
HC
Variation
Mean
SD
Within a subsample
Between subsamples
10
10
10
8
0-51
0-13
0-28
0-34
0-039
0-027
0-050
0-020
Between observers
Mean
0-66
0-22
0-39
0-52
SD
0-065
0-055
0-057
0-081
< 060
o
c 040
Runnii
1 020
Observer A
Observer B
~ Observer C
0
20
40
60
80 100 120 140 160
Number of intersections
180
c
o
(0
E
_o
o
u
04-
._
02
Rank
REFERENCES
& ROBSON, A. D. (1978). Growth of subterranean
clover in relation to the formation of endomycorrhizas by
introduced and indigenous fungi in a field soil. New Phytologist
81, 575-585.
AMBLER, J. R. & YOUNG, J. L. (1977). Techniques for determining
root length infected by vesicular-arbuscular mycorrhizae. Soil
Science Society of America Journal 41, 551-556.
BAYLIS, G . T . S. (1967). Experiments on the ecological
significance of phycomycetous mycorrhizas. New Phytologist
66, 231-243.
BETHLENFALVAY, G . J., PACOVSKY, R. S. & BROWN, M . S. (1981).
Measurement of mycorrhizal infection in soybeans. Soil Science
Society of America Journal 45, 871-875.
BiERMANN, B. & LINDERMAN, R. G (1981). Quantifying vesiculararbuscular mycorrhizae: a proposed method toward
standardization. New Phytologist 87, 63-67.
BRUNDRETT, M . C , PICHE, Y . & PETERSON, R. L . (1984). A new
method for observing the morphology of vesicular-arbuscular
mycorrhizae. Canadian Journal of Botany 62, 2128-2134.
BRUNDRETT, M . C , PICHE, Y . & PETERSON, R. L . (1985). A
developmental study of the early stages in vesicular-arbuscular
mycorrhiza formation. Canadian Journal of Botany bZ, 184194.
BuwALDA, J. G. & GoH, K. M. (1982). Host-fungus competition
for carbon as a cause of growth depressions in vesicular-arbuscular
mycorrhizal ryegrass. Soil Biology and Biochemistry 14,
103-106.
DAFT, M . J. & NICOLSON, T . H . (1972). Effect of Endogone
mycorrhiza on plant growth. IV. Quantitative relationships
between the growth of the host and the development of the
endophyte in tomato and maize. New Phytologist 71, 287-295.
DASILVA, J. A., RUIZ, E . M . & MARQUES, S. L . (1987). Individual
difTerences in magnitude estimates of inferred, remembered,
and perceived geographical distance. Bulletin of the Psychonomic
Society 25, 240-243.
DoDD, J. C. & JEFFRIES, P. (1986). Early development of
vesicular-arbuscular mycorrhizas in autumn-sown cereals. Soil
Biology and Biochemistry 18, 149-154.
ABBOTT, L . K .
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M.