Research

Is there an association between root architecture and

mycorrhizal growth response?
Hafiz Maherali
Department of Integrative Biology, University of Guelph, Guelph, ON N1G 2W1, Canada

Summary
Author for correspondence:  The symbiosis between arbuscular mycorrhizal (AM) fungi and plants is evolutionarily wide-
Hafiz Maherali spread. The response of plant growth to inoculation by these fungi (mycorrhizal growth
Tel: +1 519 824 4120 ext 52767
response; MGR) is highly variable, ranging from positive to negative. Some of this variation is
Email: maherali@uoguelph.ca
hypothesized to be associated with root structure and function. Specifically, species with a
Received: 5 May 2014 coarse root architecture, and thus a limited intrinsic capacity to absorb soil nutrients, are
Accepted: 7 June 2014
expected to derive the greatest growth benefit from inoculation with AM fungi.
 To test this hypothesis, previously published literature and phylogenetic information were
New Phytologist (2014) 204: 192–200 combined in a meta-analysis to examine the magnitude and direction of relationships among
doi: 10.1111/nph.12927 several root architectural traits and MGR.
 Published studies differed in the magnitude and direction of relationships between root

Key words: arbuscular mycorrhizal (AM)

fungi, mycorrhizal dependence, mycorrhizal
responsiveness, root diameter, root hairs,
root : shoot ratio, symbiosis.
architecture and MGR. However, when combined, the overall relationship between MGR and
allocation to roots, root diameter, root hair length and root hair density did not differ signifi-
cantly from zero.
 These findings indicate that possessing coarse roots is not necessarily a predictor of plant
growth response to AM fungal colonization. Root architecture is therefore unlikely to limit the
evolution of variation in MGR.

species with coarse root architecture, characterized by relatively

Introduction
The symbiosis between plants and arbuscular mycorrhizal (AM)
fungi is one of the most widespread trophic interactions in nature
(Brundrett, 1991; Kiers & van der Heijden, 2006). AM fungi are
an ancient lineage of obligate biotrophs in the phylum Glomer-
omycota that form associations with plants in order to obtain
energy for growth and reproduction. In return, AM fungi can
provide plants with better access to soil nutrients and improve
resistance to abiotic and biotic stressors (Newsham et al., 1995;
Smith & Read, 2008). Although the symbiosis is facultative for
many terrestrial plants, fossil evidence indicates that the relation-
ship is > 400 million yr old and nearly 75% of plant species are
estimated to form associations with AM fungi (Pirozynski &
Malloch, 1975; Remy et al., 1994; Wang & Qiu, 2006;
Brundrett, 2009). Despite the ubiquity of the AM symbiosis,
plant growth responses to AM fungi, defined as the biomass dif-
ference between a mycorrhizal plant and a non-inoculated plant
of the same species (hereafter referred to as the mycorrhizal
growth response (MGR); Janos, 2007), vary widely along a con-
tinuum from positive to negative (Johnson et al., 1997; Klirono-
mos, 2003).
The structure and function of the root system are expected to
influence how plants respond to colonization by AM fungi
(Smith & Gianinazzi-Pearson, 1988; Newsham et al., 1995;
Smith & Read, 2008). A frequently cited hypothesis is that
large diameter roots, low root hair density and short root hairs,
derive the greatest growth benefit from colonization by AM fungi
(Baylis, 1970, 1975; Smith & Gianinazzi-Pearson, 1988;
Hetrick, 1991; Newsham et al., 1995; Brundrett, 2002; Fitter,
2004; Smith & Read, 2008). This is because plants with a coarse
root architecture have limited intrinsic ability to absorb nutrients
(Bates & Lynch, 2001), and are therefore predicted to benefit
from the presence of finely structured AM fungal hyphae, which
increase the surface area available for the absorption of nutrients,
particularly phosphorus (Raven & Edwards, 2001). As originally
formulated (Baylis, 1970, 1975; Fitter, 2004), this hypothesis
proposed that a fine root architecture (specifically greater root
hair length and density) is an alternative to mycotrophy in phos-
phorus-limited soils. Therefore, root fineness is expected to be
negatively correlated with mycorrhizal dependence, defined as
‘the inability of plants without mycorrhizas to grow unless
provided with supplemental phosphorus’ (Janos, 2007, p. 88).
Although the original Baylis hypothesis was formulated for
mycorrhizal dependence, it has been tacitly extended to include
MGR, because the latter property is a general approximation of
the degree to which plants can respond to inoculation by AM
fungi (Menge et al., 1978; Hetrick, 1991; Klironomos, 2003;
Smith & Read, 2008; Hoeksema et al., 2010). The extension of
the Baylis hypothesis to MGR was also facilitated by the inter-
changeable use of mycorrhizal dependence and MGR in the

192 New Phytologist (2014) 204: 192–200 Ó 2014 The Authors

www.newphytologist.com New Phytologist Ó 2014 New Phytologist Trust
New
Phytologist
literature, even though they are functionally distinct concepts
(Janos, 2007). In addition, MGR is much more frequently
reported than mycorrhizal dependence (Hoeksema et al., 2010;
Veresoglou et al., 2012; Treseder, 2013), probably because its
estimation requires a simpler experimental design (Fitter, 2004;
Janos, 2007).
Regardless of its origin, the frequency with which relationships
between root traits and MGR are evaluated in the literature
(Menge et al., 1978; Graham & Syvertsen, 1985; Hetrick et al.,
1988; Hetrick, 1991; Smith & Read, 2008; Smith & Smith,
2011) has established the expectation that coarse roots should be
positively correlated with plant growth response to mycorrhizal
inoculation (e.g. table 4.6 in Smith & Read, 2008). Nonetheless,
there is no consensus on the direction and magnitude of the rela-
tionship between these two characteristics. Several comparative
studies with wild and agricultural species have reported associa-
tions between coarser roots, shorter root hairs and lower root hair
density and increased MGR (Hetrick et al., 1988; Manjunath &
Habte, 1991; Baon et al., 1994; Declerck et al., 1995; Schweiger
et al., 1995; Jakobsen et al., 2005). By contrast, other compara-
tive studies have indicated that root fineness, root hair density
and root hair length are either not associated or positively associ-
ated with MGR (Graham & Syvertsen, 1985; Peterson &
Farquhar, 1996; Duponnois et al., 2001; Siqueira & Saggin-
unior, 2001; Zangaro et al., 2005, 2007).
J
To determine the magnitude and direction of the overall rela-
tionship between root traits and MGR, a meta-analysis of previ-
ous studies was used to evaluate the correlation between root
traits and MGR. Based on the tacit extension of the Baylis
hypothesis to MGR (Menge et al., 1978; Hetrick, 1991), species
for which non-inoculated individuals have lower allocation to
roots and coarser root systems were expected to have higher
MGR. Specifically, decreased root : shoot ratio, low specific root
length (SRL, or the ratio of root length to root mass), thicker root
diameter, shorter root hairs and lower root hair frequency were
predicted to associate with increased MGR across seed plants.
Because of shared ancestry, species are not statistically indepen-
dent, and this lack of independence could either produce spuri-
ous correlations or obscure legitimate correlations (Felsenstein,
1985). To account for this effect, interspecific correlations
between root traits and MGR in previously published studies
were re-calculated by formally taking into account the phyloge-
netic relationships among species in the sample. These phyloge-
netically corrected correlations and their associated error
variances were used in the meta-analysis.
Materials and Methods
Data collection
To examine the relationships between root traits and the growth
response of plants to AM fungi, Web of Science and Google
Scholar were searched using arbuscular, mycorrhizal, mycorrhiza,
mycorrhizae, fungal, fungi, AM fungal, AM fungi, AMF and
each root trait (specific root length, fine root diameter, root hair
length and root hair density) as terms. The results of these
Ó 2014 The Authors
New Phytologist Ó 2014 New Phytologist Trust
Research 193
searches were screened to determine whether they contained
information on quantitative measurements of both a root trait
under non-mycorrhizal conditions, an assessment of MGR via
direct inoculation with AM fungi, and whether the number of
species in the sample was ≥ 4. This level of replication within a
study was required in order to calculate the correlation coefficient
and its associated error variance for meta-analysis.
From each study, data on plant biomass were obtained for
both the AM fungal treatment and the non-inoculated treatment
or the ratio of biomass from the two treatments. In addition,
information on at least one of SRL, root diameter, root hair
length and root hair density was recorded from plants in the non-
inoculated treatment. In cases in which SRL was not reported
directly, it was calculated as the ratio of root length and root mass
if these values were reported. High SRL values typically indicate
thinner roots with greater total surface area for nutrient absorp-
tion, whereas low SRL values indicate thicker roots with lower
total surface area (Craine et al., 2001). Root : shoot ratios were
recorded when reported, or calculated from biomass data.
Because not all studies reported all root traits, sample sizes for
each relationship differed (Table 1).
Quantifying MGR
To quantify MGR, the effect size of the AM fungal treatment on
dry plant biomass (either shoot biomass or total biomass,
depending on the study) was calculated as the log response ratio,
MGR = loge[Xi/Xn], where Xi is the inoculated biomass and Xn is
the non-inoculated biomass (Hoeksema et al., 2010). Positive
MGR values indicate that plant biomass increased in response to
inoculation, whereas negative values indicate that plant biomass
decreased in response to inoculation. This metric is preferred over
previous formulations of MGR (reviewed in Janos, 2007) because
values are not bounded by zero, and its statistical properties are
appropriate for meta-analyses (Hedges et al., 1999). In situations
in which plants were grown with multiple AM fungal inocula
treatments, the response ratio was calculated by averaging Xi
across treatments, as described by van der Heijden et al. (1998).
In situations in which other calculations of MGR were reported,
they were converted to log response ratios using the formulae
described by Allison & Goldberg (2002). In situations in which
plants were grown across a gradient of soil phosphorus, maxi-
mum MGR values were recorded, and plant traits were recorded
from the corresponding non-mycorrhizal treatment.
Meta-analysis of relationships between root traits and
MGR
A random effects meta-analysis (Borenstein et al., 2009) was used
to examine the overall magnitude and direction of associations
between the root architectural traits of non-inoculated plants and
MGR. A random effects analysis was used because the overall
effect size is calculated assuming that variation among studies can
occur because of differences in experimental conditions. This is
applicable to the present meta-analysis because studies differed in
terms of plant species identity, growth conditions and sources of
New Phytologist (2014) 204: 192–200
www.newphytologist.com
 New
194 Research Phytologist

mycorrhizal inoculum. By assuming that these factors vary

relationship between variation in the regression residuals and phylogeny and, when k = 1, there is complete dependence between residual variation and a Brownian model of evolution along the phy-
Root hair
density

For each relationship, the maximum likelihood estimate of k, or the degree to which residuals were associated with the phylogenetic variance–covariance matrix, are shown. When k = 0, there is no
among studies, a random effects meta-analysis is generalizable to

0.92

0.44
0.5
Table 1 A summary of the growth form, biome and sample sizes of 12 studies that report relationships between at least one of five root architectural traits and the mycorrhizal growth response

the overall hypothesis (Borenstein et al., 2009).

–
–

–
–

–
0
0

0
Before conducting the meta-analysis, the within-study Pearson
correlations between root architectural traits and MGR, and their
Root hair
length

0.86 associated variances, were calculated. The correlation coefficient

0.56

logeny. Study abbreviations match those on the x axes of Figs 1 and 2. Soil extractable phosphorus (P), method of P extraction, growth form and biome information are also listed.
0.5

was used because it represents a standardized effect size, which

1
0

0
–
–

–
–

–
allows the effect sizes among studies to be expressed on the same
scale. To account for the effect of shared ancestry in the estimate
diameter

of correlation coefficients from the original studies, correlation

Root

0.91
0.36

0.34
coefficients within each study were calculated from a phyloge-
–
–

–
–
0
0

1
netic generalized least-squares (PGLS) regression, where the phy-
SRL

logenetic variance–covariance matrix is incorporated directly into

1

0
0
1

1
–
–

–
–

–
–

– the calculation of effect size (Martins & Hansen, 1997; Pagel,

1999). To facilitate PGLS regression, a phylogenetic tree for all
Root : shoot

taxa in the meta-analysis was derived from the Angiosperm

Phylogeny Group seed plant phylogeny reference tree (APG III;
ratio

0.5

Stevens, 2001) which is contained within Phylomatic (http://

1

0
1
0

1
0
–

–
–

–
–
k

phylodiversity.net/phylomatic/; Webb et al., 2008). The resulting

tree (Supporting Information Fig. S1) was assigned branch
17

13

29
22
7

4
8
4
4
5

782
n

lengths using node ages from Wikstrom et al. (2001) and the
‘bladj’ function in Phylomatic.
Subtropical
Temperate
Temperate

Temperate
Temperate

Temperate

To estimate the correlation coefficient in each study, the phy-

Tropical
Tropical

Tropical
Tropical

Tropical

Tropical
Biome

logenetic tree was pruned to create unique trees for each study,
depending on the identity of the species present. To quantify the
degree of dependence on phylogeny for each study, a scaling fac-
tor k was estimated using maximum likelihood. k was unique to
Growth form

Herb, grass
Herb, grass

Herb, grass

Herb, grass
Herb, tree

each study because it depends on the identity of the species sam-

pled, and ranges from zero, indicating complete independence
Herb

Tree

Tree
Tree
Tree

Tree

Tree

between variation in the regression residuals and phylogeny, to

unity, indicating complete dependence between residual variation
and a Brownian model of evolution along the phylogeny
0.02 mg l 1 (method unknown)

(Freckleton et al., 2002). When k = 0, the PGLS regression is

identical to ordinary least-squares (OLS) regression. Estimates of
1.8 mg kg 1 (Mehlich I)

k were performed in R version 3.02 (R Core Team, 2013) and

3 mg kg 1 (Mehlich I)

4.82 mg kg 1 (Bray I)

n = 77 for root diameter and root hair density; n = 72 for root hair length.

the ‘pgls’ command in the package caper, version 0.5.2 (Orme

2 mg kg 1 (Olsen)
Extractable soil P

7 lg g 1 (Olsen)
4.8 ppm (Olsen)

10 ppm (Bray I)

20 ppm (Bray I)

et al., 2013).
To carry out the random effects meta-analysis for each root
Fertilized

Fertilized
Fertilized

Included data from a companion paper: Zangaro et al. (2003).

architectural trait, the correlations between traits and MGR, and

their variances, were combined using the equations and method-
ology described in Borenstein et al. (2009). Effect sizes from indi-
vidual studies were weighted by the inverse of their variances
Abbreviation

before being combined in the meta-analysis. To determine

whether the inclusion of a particular study influenced the overall
MAN
OSO
GRA
DUP

POP

ZAN
DEC

SCH

effect size, a sensitivity analysis was carried out. This entailed the
HET

STA
SIQ
HIL

iterative removal of one study and the re-calculation of the overall

effect size of the meta-analysis with the remaining studies. A two-
unior (2001)

tailed z-test was used to determine whether the overall effect size
Graham & Syvertsen (1985)

Manjunath & Habte (1991)

for each meta-analysis differed from zero (Borenstein et al.,

Duponnois et al. (2001)

2009).
Schweiger et al. (1995)

Zangaro et al. (2005)1

Declerck et al. (1995)

Osonubi et al. (1991)

Siqueira & Saggin-J

Hetrick et al. (1988)

Pope et al. (1983)

Stanescu (2012)
Hill et al. (2010)

Results
The dataset for analysis was compiled from 12 studies (Table 1)
(MGR)

Study

comprising 196 experimental trials (Table S1). Studies were per-

formed on plants ranging from 28 to 262 d old and growing in
1

New Phytologist (2014) 204: 192–200 Ó 2014 The Authors

www.newphytologist.com New Phytologist Ó 2014 New Phytologist Trust
New
Phytologist
pots in glasshouses or other controlled environments. The experi-
mental trials contained representatives of 40 seed plant families.
A plurality of observations came from the Fabaceae, which repre-
sented 73 of the 196 trials. Phylogeny was correlated with regres-
sion residuals in a majority of studies (Table 1). Of the 33
reported correlations between root architectural traits and MGR,
k was > 0 in 18 cases. Only one study (Manjunath & Habte,
1991) lacked phylogenetic influence on the calculation of correla-
tions.
Root allocation and aspects of root fineness were not associated
with MGR in the meta-analysis. The root : shoot ratio was posi-
tively correlated with MGR in the study by Zangaro et al. (2005),
but no other study contained a statistically significant relation-
ship (Fig. 1a) and the overall effect size did not differ significantly
from zero (r = 0.26, P = 0.32). The overall effect size was insensi-
tive to the iterative removal of studies with one exception. When
the study by Stanescu (2012) was removed, the root : shoot ratio
(a)
(b)
Fig. 1 Pearson correlation coefficients for (c)
each study (closed circles), the overall effect
size (open triangles) in the meta-analysis (a–
c) and the sensitivity of the overall effect size
to the iterative removal of single studies (d–
f). Relationships are between root : shoot
ratio (a, d), specific root length (SRL) (b, e)
and root diameter (c, f) of non-inoculated
plants and mycorrhizal growth response
(MGR). Study abbreviation codes correspond
to those listed in Table 1. Vertical lines
around the symbols represent the 95%
confidence intervals. Correlations that
differed significantly from zero are indicated:
*, P < 0.05; **, P < 0.01; ***, P < 0.001.
Ó 2014 The Authors
New Phytologist Ó 2014 New Phytologist Trust
Research 195
was positively correlated with MGR (Fig. 1d; r = 0.44,
P = 0.047). SRL was negatively correlated with MGR in three of
five studies (Pope et al., 1983; Graham & Syvertsen, 1985;
Manjunath & Habte, 1991), but the overall effect size did not
differ from zero (Fig. 1b; r = 0.35, P = 0.22). The overall effect
size for the relationship between SRL and MGR remained non-
significant in the sensitivity analysis (Fig. 1e). Root diameter was
positively correlated with MGR in the studies by Hetrick et al.
(1988) and Manjunath & Habte (1991), but negatively corre-
lated with MGR in the studies by Schweiger et al. (1995) and
Zangaro et al. (2005), but the overall effect size did not differ
from zero (Fig. 1c, r = 0.048, P = 0.81). Similarly, the relation-
ship between root diameter and MGR remained non-significant
in the sensitivity analysis (Fig. 1f).
Root hair dimensions and frequency were also not associated
with MGR in the meta-analysis. Root hair length was negatively
correlated with MGR in the studies by Declerck et al. (1995) and
(d)
(e)
(f)
New Phytologist (2014) 204: 192–200
www.newphytologist.com

196 Research
Manjunath & Habte (1991), but positively correlated with MGR
in the study by Siqueira & Saggin-J unior (2001) (Fig. 2a). Never-
theless, the overall effect size did not differ from zero (r = 0.029,
P = 0.90) and this outcome was consistently observed in the sen-
sitivity analysis (Fig. 2c). Root hair density was negatively corre-
lated with MGR in the studies by Declerck et al. (1995) and
Manjunath & Habte (1991), but positively correlated with MGR
in the studies by Hill et al. (2010) and Zangaro et al. (2005)
(Fig. 2b). However, the overall effect size did not differ from zero
(r = 0.057, P = 0.81), an outcome that was also consistently
observed in the sensitivity analysis (Fig. 2d).
Discussion
Meta-analyses of the relationship between root traits and MGR
did not support the prediction that coarse root architecture is
associated with greater plant growth benefit from AM fungi
(Menge et al., 1978; Hetrick et al., 1988; Hetrick, 1991; Manj-
unath & Habte, 1991; Brundrett, 2002; Smith & Read, 2008).
The overall effect sizes of the relationships between MGR and
root : shoot ratio, SRL, root diameter, root hair length and root
hair density did not differ from zero (Figs 1a–c, 2a,b). The out-
come of the meta-analyses was not generally sensitive to the
inclusion of a particular study (Figs 1d–f, 2c,d). Specifically, the
outcome was not influenced by the exclusion of studies with large
sample sizes, and thus high leverage (Zangaro et al., 2005;
Table 1). The only exception to this pattern occurred for the rela-
tionship between the root : shoot ratio and MGR when the study
by Stanescu (2012) was excluded. However, the overall effect size
(a) (c)
(b) (d)
New Phytologist (2014) 204: 192–200
www.newphytologist.com
New
Phytologist
in this case was the opposite of expectations – increased
root : shoot ratio in non-mycorrhizal plants was positively associ-
ated with MGR. These results suggest that there is no broad sup-
port for the hypothesis that species with a coarse root architecture
are more likely to derive the greatest growth benefit from coloni-
zation by AM fungi.
Although there were no overall associations between root
architectural traits and MGR in the meta-analysis, some of the
contributing studies had significant effect sizes. The pattern of
variation among contributing studies, or lack thereof, suggested
three ways in which a non-significant overall effect size occurred.
First, for the root : shoot ratio (Fig. 1a), there were generally weak
relationships in the contributing studies, which resulted in a non-
significant overall effect size. The single study for which there was
a statistically significant relationship (Zangaro et al., 2005) did
not have sufficient weight to counteract the influence of the
majority of studies which showed no significant relationship. Sec-
ond, for root diameter, root hair length and root hair density
(Figs. 1c, 2a,b), there were approximately equal proportions of
significant positive and negative relationships among the contrib-
uting studies, and these opposite effects averaged out to produce
a non-significant overall effect. The tendency for equal propor-
tions of studies to fall on opposite sides of the weighted mean
effect is not unusual in meta-analyses, and suggests that sampling
error could have been responsible for differences between studies
(Palmer, 2000). Third, for SRL, significant correlations were
consistently negative, yet did not produce an overall negative
effect size (Fig. 1b). Although the overall weighted correlation
was the strongest among the root traits (r = 0.35), this
Fig. 2 Pearson correlation coefficients for
each study (closed circles), the overall effect
size (open triangles) in the meta-analysis (a,
b) and the sensitivity of the overall effect size
to the iterative removal of single studies (c,
d). Relationships are between root hair
length (a, c) and root hair density (b, d) of
non-inoculated plants and mycorrhizal
growth response (MGR). Study abbreviations
correspond to those listed in Table 1. Vertical
lines around the symbols represent the 95%
confidence intervals. Correlations that
differed significantly from zero are indicated:
*, P < 0.05; **, P < 0.01; ***, P < 0.001.
Ó 2014 The Authors
New Phytologist Ó 2014 New Phytologist Trust
New
Phytologist
meta-analysis had the smallest sample size (Table 1), which led to
high analysis-wide variance. Thus, it is likely that the SRL
meta-analysis lacked the necessary power to reach conclusions
about the nature of the relationship between SRL and MGR.
Variation in the magnitude and direction of effect sizes in con-
tributing studies could potentially be associated with covariates,
such as growth form, biome/habitat and soil phosphorus content
(Table 1). Although there were an insufficient number of studies
within each meta-analysis to statistically test for the influence of
covariates, a comparison of the covariates among studies suggests
that it is unlikely that many of these factors were responsible for
effect size variation. This is because similar effect sizes were often
found for studies that differed in growth form and habitat. For
example, there were significant positive relationships between
root diameter and MGR in Hetrick et al. (1988) and Manjunath
& Habte (1991), but the former study was performed with tem-
perate herbs and grasses, whereas the latter was performed with
tropical trees (Fig. 1c). Similarly, there were significant negative
relationships between root diameter and MGR in Schweiger et al.
(1995) and Zangaro et al. (2005), even though the former was
performed with temperate herbs and grasses and the latter with
tropical trees. These types of patterns were also observed for root
hair length and root hair density (Fig. 2a,b). Differences in phos-
phorus availability could have influenced both the morphology
of root systems of non-inoculated plants (Raven & Edwards,
2001) and the magnitude of MGR (Hoeksema et al., 2010), and
potentially contributed to variation in the magnitude and direc-
tion of correlations. Although extractable phosphorus in soil was
reported in a majority of studies, differences in the analysis
method among studies meant that values could not be compared
on an absolute scale (Wolf & Baker, 1985). Therefore, it is not
known whether and by how much phosphorus availability in soil
influenced inter-study variability in relationships between root
architecture and MGR.
Although a relationship between root architecture and MGR
was not found in the present meta-analysis, it should be noted
that there are other biologically meaningful aspects of root system
architecture and plant growth response to AM fungi that were
not captured in the analysis. First, the present study used fre-
quently reported root architectural traits, but other less studied
aspects of the root system, such as branching frequency (Berta
et al., 1995), could be associated with MGR. Second, it is possi-
ble that trade-offs between root traits influenced the outcome.
For example, plants may enhance nutrient absorption by increas-
ing SRL, root hair length or root hair density, but possibly not all
three simultaneously. If alternative strategies for producing fine
root architecture are employed by different species (Siqueira &
Saggin-J unior, 2001), the detection of a correlation between root
architecture and MGR could be highly dependent on the sample
of species used. Similarly, a relationship between root architec-
ture and MGR could be detected if different root architectural
metrics could be combined to reflect whole root system function.
Finally, the growth response of plants to AM fungi depends on
the identity of both the plant and fungal partner (Klironomos,
2003). Therefore, it is possible that a relationship between root
architecture and MGR could exist if studies had matched plant
Ó 2014 The Authors
New Phytologist Ó 2014 New Phytologist Trust
Research 197
species with the AM fungal partners that most positively influ-
enced plant growth.
One explanation for the independence between root architec-
tural traits and MGR is the ability of plants to respond plastically
to fungal colonization. For example, several studies have sug-
gested that inoculation with AM fungi causes plants to produce
coarser root systems, which could be a coordinated response that
facilitates a shift from root-driven to mycorrhizae-driven nutrient
uptake (Hetrick, 1991; Berta et al., 1995; Hooker & Atkinson,
1996; Zangaro et al., 2007). This possibility could not be tested
in the present meta-analysis because only a small number of stud-
ies reported the root architecture of both non-inoculated and
inoculated plants (Osonubi et al., 1991; Zangaro et al., 2005).
Although root plasticity to AM fungal colonization is a potential
explanation for a lack of relationship between root architecture
and MGR, plants do not always produce coarser roots following
inoculation by AM fungi (Schroeder & Janos, 2005; Zangaro
et al., 2005; Wu et al., 2010, 2011). Variability in the outcome
of studies that evaluate the plasticity of root architecture to fungal
colonization may arise from a lack of control for the effects of
allometric scaling between root architecture and plant size. For
example, a common plastic response to AM fungal inoculation is
reduced root : shoot ratio (Veresoglou et al., 2012). However,
root : shoot ratio is also negatively correlated with size and devel-
opment (McConnaughay & Coleman, 1999). Thus, mycorrhizal
plants may have a low root : shoot ratio because they are larger,
and not because of a direct effect of AM fungal inoculation
(Veresoglou et al., 2012). The potential for size dependence as a
complicating factor in the determination of whether fungal
inoculation produces coarse roots has not been explored experi-
mentally (Veresoglou et al., 2012). Therefore, future studies of
the effects of AM fungal inoculation on root architecture should
be performed by comparing inoculated and non-inoculated
plants in an ontologically controlled manner (McConnaughay &
Coleman, 1999). Such data are necessary to test whether root
plasticity to AM fungi is a common feature of plants and
influences the outcome of relationships between root traits and
MGR.
Even though expectations for a relationship between root
architecture and MGR were derived from theories of mycorrhizal
dependence, the lack of a relationship does not preclude the exis-
tence of a correlation between root architecture and mycorrhizal
dependence (Baylis, 1970, 1975; Fitter, 2004). This is because
MGR and mycorrhizal dependence are not quantitatively equiva-
lent (Janos, 2007). For example, species that are dependent on
AM fungi will necessarily have a positive MGR, whereas species
that are not dependent on AM fungi could have MGRs anywhere
along a continuum from negative to positive (Sawers et al.,
2008). Thus, if there is a relationship between root architecture
and mycorrhizal dependence, it was unlikely to be detected in the
present analysis because species with different levels of mycorrhi-
zal dependence could theoretically have the same MGR. As has
been stated elsewhere (Fitter, 2004; Janos, 2007), mycorrhizal
dependence is less frequently quantified than MGR. Moreover,
the lack of overlap in these metrics in published studies not only
prevents broad tests of the hypothesis between root architecture
New Phytologist (2014) 204: 192–200
www.newphytologist.com

198 Research
and mycorrhizal dependence, but also makes it impossible to par-
tition out dependence and non-dependence components from
MGR (Sawers et al., 2008). Thus, one recommendation from the
present study is to re-iterate the call for researchers to quantify
both MGR and mycorrhizal dependence in the same study using
standard methods (Janos, 2007; Sawers et al., 2008).
In conclusion, the findings of this meta-analysis indicate that
there is no broad support for the expectation that species with
a coarse root architecture have higher MGR. Therefore, the
tacit extension of the Baylis hypothesis to MGR is not war-
ranted. More generally, these findings also suggest that root
architecture need not limit the evolution of interspecific vari-
ability in plant growth responses to AM fungal inoculation
(Hoeksema et al., 2010). AM fungi are now known to influ-
ence many aspects of plant growth, structure and function
beyond the acquisition of phosphorus, including the uptake of
other elements and water, protection from pathogens and her-
bivores, and resistance to toxic substances and environmental
stressors (Newsham et al., 1995; Powell et al., 2009; Sikes et al.,
2010; Chagnon et al., 2013). Furthermore, plant growth
responses to AM fungi can be influenced by cost–benefit ratios
of resource exchange between the partners, soil resource avail-
ability, climatic factors and competition with other plants and
fungi (Smith & Gianinazzi-Pearson, 1988; Klironomos, 2003;
Maherali & Klironomos, 2007; Bever et al., 2009; Hoeksema
et al., 2010; Johnson, 2010; Kiers et al., 2011; Reinhart et al.,
2012). Thus, the magnitude and direction of plant growth
responses to AM fungal inoculation could be influenced by
many different ecological factors, all of which could act as
agents of selection on the symbiosis. Because of this, the causes
of variation in the mycorrhizal symbiosis should be tested
experimentally by quantifying the strength and direction of nat-
ural selection on the symbioses, whilst, at the same time,
manipulating putative ecological agents of selection. Such stud-
ies have been carried out for other types of biotic interactions
(Weis & Gorman, 1990; Juenger & Bergelson, 2000; Heath &
Tiffin, 2007) and have great potential to unravel the causes of
variation in the interactions between plants and AM fungi
(Hoeksema, 2010).
Acknowledgements
This work was supported by a Discovery grant from the Natural
Sciences and Engineering Research Council (NSERC), the Uni-
versity of Guelph and sabbatical support from the National Evo-
lutionary Synthesis Center (NESCent), NSF #EF-0423641. I
thank C. M. Caruso, A. H. Fitter and anonymous reviewers for
comments that improved the manuscript. I also thank the schol-
arly community and administrative staff at NESCent for provid-
ing a superb environment for synthesis research.
References
Allison VJ, Goldberg D. 2002. Species-level versus community-level patterns of
mycorrhizal dependence on phosphorus: an example of Simpson’s paradox.
Functional Ecology 16: 346–352.
New Phytologist (2014) 204: 192–200
www.newphytologist.com
New
Phytologist
Baon JB, Smith SE, Alston AM. 1994. Growth response and phosphorus uptake
of rye with long and short root hairs: interactions with mycorrhizal infection.
Plant and Soil 167: 247–254.
Bates TR, Lynch JP. 2001. Root hairs confer a competitive advantage under low
phosphorus availability. Plant and Soil 236: 243–250.
Baylis GTS. 1970. Root hairs and phycomycetous mycorrhizas in phosphorus
deficient soil. Plant and Soil 33: 713–716.
Baylis GTS. 1975. The magnolioid mycorrhiza and mycotrophy in root systems
derived from it. In: Sanders FE, Mosse B, Tinker PB, eds. Endomycorrhizas.
New York, NY, USA: Academic Press, 373–389.
Berta G, Trotta A, Fusconi A, Hooker JE, Munro M, Atkinson D, Giovannetti
M, Morini S, Fortuna P, Tisserant B et al. 1995. Arbuscular mycorrhizal
induced changes to plant growth and root system morphology in Prunus
cerasifera. Tree Physiology 15: 281–293.
Bever JD, Richardson SC, Lawrence BM, Holmes J, Watson M. 2009.
Preferential allocation to beneficial symbiont with spatial structure maintains
mycorrhizal mutualism. Ecology Letters 12: 13–21.
Borenstein M, Hedges LV, Higgins JPT, Rothstein HR. 2009. Introduction to
meta-analysis. Chichester, UK: Wiley.
Brundrett MC. 1991. Mycorrhizas in natural ecosystems. Advances in Ecological
Research 21: 171–313.
Brundrett MC. 2002. Coevolution of roots and mycorrhizas of land plants. New
Phytologist 154: 275–304.
Brundrett MC. 2009. Mycorrhizal associations and other means of nutrition of
vascular plants: understanding the global diversity of host plants by resolving
conflicting information and developing reliable means of diagnosis. Plant and
Soil 320: 37–77.
Chagnon PL, Bradley RL, Maherali H, Klironomos JN. 2013. A trait-based
framework to understand life history of mycorrhizal fungi. Trends in Plant
Science 18: 484–491.
Craine JM, Froehle J, Tilman DG, Wedin DA, Chapin FS III. 2001. The
relationships among root and leaf traits of 76 grassland species and relative
abundance along fertility and disturbance gradients. Oikos 93: 274–285.
Declerck S, Plenchette C, Strullu DG. 1995. Mycorrhizal dependency of banana
(Musa acuminata, AAA group) cultivar. Plant and Soil 176: 183–187.
Duponnois R, Plenchette C, Ba AM. 2001. Growth stimulation of seventeen
fallow leguminous plants inoculated with Glomus aggregatum in Senegal.
European Journal of Soil Biology 37: 181–186.
Felsenstein J. 1985. Phylogenies and the comparative method. American
Naturalist 125: 1–15.
Fitter AH. 2004. Magnolioid roots – hairs, architecture and mycorrhizal
dependency. New Phytologist 164: 15–16.
Freckleton RP, Harvey PH, Pagel M. 2002. Phylogenetic analysis and comparative
data: a test and review of evidence. American Naturalist 160: 712–726.
Graham JH, Syvertsen J. 1985. Host determinants of mycorrhizal dependency of
citrus rootstock seedlings. New Phytologist 101: 667–676.
Heath KD, Tiffin P. 2007. Context-dependence in the coevolution of plant and
rhizobium mutualists. Proceedings of the Royal Society B: Biological Sciences 274:
1905–1912.
Hedges LV, Gurevitch J, Curtis PS. 1999. The meta-analysis of response ratios
in experimental ecology. Ecology 80: 1150–1156.
van der Heijden MGA, Boller T, Wiemken A, Sanders IR. 1998. Different
arbuscular mycorrhizal fungal species are potential determinants of plant
community structure. Ecology 79: 2082–2091.
Hetrick BAD. 1991. Mycorrhizas and root architecture. Experentia 47: 355–362.
Hetrick BAD, Kitt DG, Wilson GT. 1988. Mycorrhizal dependence and growth
habit of warm-season and cool-season tallgrass prairie plants. Canadian Journal
of Botany 66: 1376–1380.
Hill JO, Simpson RJ, Ryan MH. 2010. Root hair morphology and mycorrhizal
colonisation of pasture species in response to phosphorus and nitrogen
nutrition. Crop and Pasture Science 61: 122–131.
Hoeksema JD. 2010. Ongoing coevolution in mycorrhizal interactions. New
Phytologist 187: 286–300.
Hoeksema JD, Chaudhary VB, Gehring CA, Johnson NC, Karst J, Koide RT,
Pringle A, Zabinski C, Bever JD, Moore JC et al. 2010. A meta-analysis of
context-dependency in plant response to inoculation with mycorrhizal fungi.
Ecology Letters 13: 394–407.
Ó 2014 The Authors
New Phytologist Ó 2014 New Phytologist Trust
New
Phytologist
Hooker JE, Atkinson D. 1996. Arbuscular mycorrhizal fungi-induced alteration
to tree-root architecture and longevity. Zeitschrift fur Pflanzenernahrung und
Bodenkunde 234: 229–234.
Jakobsen I, Chen B, Munkvold L, Lundsgaard T, Zhu Y-G. 2005. Contrasting
phosphate acquisition of mycorrhizal fungi with that of root hairs using the
root hairless barley mutant. Plant, Cell & Environment 28: 928–938.
Janos DP. 2007. Plant responsiveness to mycorrhizas differs from dependence
upon mycorrhizas. Mycorrhiza 17: 75–91.
Johnson NC. 2010. Resource stoichiometry elucidates the structure and function
of arbuscular mycorrhizas across scales. New Phytologist 185: 631–647.
Johnson NC, Graham JH, Smith FA. 1997. Functioning of mycorrhizal
associations along the mutualism–parasitism continuum. New Phytologist 135:
575–586.
Juenger T, Bergelson J. 2000. The evolution of compensation to herbivory in
scarlet gilia, Ipomopsis aggregata; herbivore-imposed natural selection and the
quantitative genetics of tolerance. Evolution 54: 764–777.
Kiers ET, Duhamel M, Beesetty Y, Mensah J, Franken O, Verbruggen E,
Fellbaum CR, Kowalchuk G, Hart MM, Bago A et al. 2011. Reciprocal rewards
stabilize cooperation in the mycorrhizal symbiosis. Science 333: 880–882.
Kiers ET, van der Heijden M. 2006. Mutualistic stability in the arbuscular
mycorrhizal symbiosis: exploring hypotheses of evolutionary cooperation.
Ecology 87: 1627–1636.
Klironomos JN. 2003. Variation in plant response to native and exotic arbuscular
mycorrhizal fungi. Ecology 84: 2292–2301.
Maherali H, Klironomos JN. 2007. Influence of phylogeny on fungal
community assembly and ecosystem functioning. Science 316: 1746–1748.
Manjunath A, Habte M. 1991. Root morphological characteristics of host species
having distinct mycorrhizal dependency. Canadian Journal of Botany 69:
671–676.
Martins EP, Hansen TF. 1997. Phylogenies and the comparative method: a
general approach to incorporating phylogenetic information into the analysis of
inter-specific data. American Naturalist 149: 646–667.
McConnaughay KDM, Coleman JS. 1999. Biomass allocation in plants:
ontogeny or optimality? A test along three resource gradients. Ecology 80:
2581–2593.
Menge JA, Johnson ELV, Platt RG. 1978. Mycorrhizal dependency of several
citrus cultivars under three nutrient regimes. New Phytologist 81: 553–559.
Newsham KK, Fitter AH, Watkinson AR. 1995. Multi-functionality and
biodiversity in arbuscular mycorrhizas. Trends in Ecology & Evolution 10:
407–411.
Orme D, Freckleton R, Thomas G, Petzoldt T, Fritz S, Isaac N, Pearse W.
2013. caper: Comparative Analyses of Phylogenetics and Evolution in R. R package
version 0.5.2. [WWW document] URL http://CRAN.R-project.org/
package=caper [accessed 6 March 2014].
Osonubi O, Mulongoy K, Awotoye OO, Atayese MO, Okali DUU. 1991. Effects
of ectomycorrhizal and vesicular–arbuscular mycorrhizal fungi on drought
tolerance of four leguminous woody seedlings. Plant and Soil 136: 131–143.
Pagel M. 1999. Inferring the historical patterns of biological evolution. Nature
401: 877–884.
Palmer AR. 2000. Quasireplication and the contract of error: lessons from sex
ratios, heritabilities and fluctuating asymmetry. Annual Review of Ecology and
Systematics 31: 441–480.
Peterson RL, Farquhar ML. 1996. Root hairs: specialized tubular cells extending
root surfaces. Botanical Review 62: 1–40.
Pirozynski KA, Malloch DW. 1975. The origin of land plants: a matter of
mycotropism. Biosystems 6: 153–164.
Pope PE, Chaney WR, Rhodes JD. 1983. The mycorrhizal dependency of four
hardwood tree species. Canadian Journal of Botany 61: 412–417.
Powell JR, Parrent JL, Klironomos JN, Hart MM, Rillig MC, Maherali H.
2009. Phylogenetic trait conservatism and the evolution of functional tradeoffs
in arbuscular mycorrhizal fungi. Proceedings of the Royal Society B: Biological
Sciences 276: 4237–4245.
R Core Team. 2013. R: A language and environment for statistical computing. R
Foundation for Statistical Computing, Vienna, Austria. [WWW document]
URL http://www.R-project.org/ [accessed 6 March 2014].
Raven JA, Edwards D. 2001. Roots: evolutionary origins and biogeochemical
significance. Journal of Experimental Botany 52: 381–401.
Ó 2014 The Authors
New Phytologist Ó 2014 New Phytologist Trust
Research 199
Reinhart KO, Wilson GWT, Rinella MJ. 2012. Predicting plant responses to
mycorrhizae: integrating evolutionary history and plant traits. Ecology Letters
15: 689–695.
Remy W, Taylor TN, Hass H. 1994. Four hundred-million-year-old vesicular
arbuscular mycorrhizae. Proceedings of the National Academy of Sciences, USA
91: 11841–11843.
Sawers RJH, Gutjahr C, Paszkowski U. 2008. Cereal mycorrhiza: an ancient
symbiosis in modern agriculture. Trends in Plant Sciences 13: 93–97.
Schroeder MS, Janos DP. 2005. Plant growth, phosphorus nutrition, and root
morphological responses to arbuscular mycorrhizas, phosphorus fertilization,
and intraspecific density. Mycorrhiza 15: 203–216.
Schweiger PF, Robson AD, Barrow NJ. 1995. Root hair length determines
beneficial effect of a Glomus species on shoot growth of some pasture species.
New Phytologist 131: 247–254.
Sikes BA, Powell JR, Rillig MC. 2010. Deciphering the relative contribution of
multiple functions within plant–microbe symbioses. Ecology 91: 1591–1597.
Siqueira J, Saggin-J unior O. 2001. Dependency on arbuscular mycorrhizal fungi
and responsiveness of some Brazilian native woody species. Mycorrhiza 11:
245–255.
Smith SE, Gianinazzi-Pearson V. 1988. Physiological interactions between
symbionts in vesicular–arbuscular mycorrhizal plants. Annual Review of Plant
Physiology and Plant Molecular Biology 39: 221–244.
Smith SE, Read DJ. 2008. Mycorrhizal symbiosis. New York, NY, USA: Academic
Press.
Smith SE, Smith FA. 2011. Roles of arbuscular mycorrhizas in plant nutrition
and growth: new paradigms from cellular to ecosystem scales. Annual Review of
Plant Biology 62: 227–250.
Stanescu S. 2012. Effects of mycorrhizae and soil biota feedback on the outcome of
plant competition. MSc thesis, University of Guelph, Guelph, ON, Canada.
Stevens PF. 2001. Angiosperm phylogeny website. [WWW document] URL http://
www.mobot.org/MOBOT/research/APweb/ [accessed 3 March 2014].
Treseder KK. 2013. Marschner Review: the extent of mycorrhizal colonization of
roots and its influence on plant growth and phosphorus content. Plant and Soil
371: 1–13.
Veresoglou SD, Menexes G, Rillig MC. 2012. Do arbuscular mycorrhizal fungi
affect the allometric partition of host plant biomass to shoots and roots? A
meta-analysis of studies from 1990 to 2010. Mycorrhiza 22: 227–235.
Wang B, Qiu Y-L. 2006. Phylogenetic distribution and evolution of mycorrhizas
in land plants. Mycorrhiza 16: 299–363.
Webb CO, Ackerly DD, Kembel SW. 2008. Phylocom: software for the analysis
of phylogenetic community structure and trait evolution. Bioinformatics 24:
2098–2100.
Weis AE, Gorman WL. 1990. Measuring selection on reaction norms:
explorations of the Solidago–Eurosta system. Evolution 44: 820–831.
Wikstrom N, Savolainen V, Chase MW. 2001. Evolution of angiosperms:
calibrating the family tree. Proceedings of the Royal Society B: Biological Sciences
268: 2211–2220.
Wolf AM, Baker DE. 1985. Comparisons of soil test phosphorus by Olsen, Bray
P1, Mehlich-I, and Mehlich-3 Soil Test. Communications in Soil Science &
Plant Analysis 16: 467–484.
Wu Q-S, Zou Y-N, He X-H. 2010. Contributions of arbuscular mycorrhizal
fungi to growth, photosynthesis, root morphology and ionic balance of citrus
seedlings under salt stress. Acta Physiologiae Plantarum 32: 297–304.
Wu Q-S, Zou Y-N, He X-H, Luo P. 2011. Arbuscular mycorrhizal fungi can
alter some root characters and physiological status in trifoliate orange (Poncirus
trifoliata L. Raf.) seedlings. Plant Growth Regulation 65: 273–278.
Zangaro W, Nishidate FR, Camargo FRS, Romagnoli GG, Vandressen J. 2005.
Relationships among arbuscular mycorrhizas, root morphology and seedling
growth of tropical native woody species in southern Brazil. Journal of Tropical
Ecology 21: 529–540.
Zangaro W, Nishidate FR, Vandresen J, Andrade G, Nogueira MA. 2007. Root
mycorrhizal colonization and plant responsiveness are related to root plasticity,
soil fertility and successional status of native woody species in southern Brazil.
Journal of Tropical Ecology 23: 53–62.
Zangaro W, Nisizaki SMA, Domingos JCB, Nakano EM. 2003. Mycorrhizal
response and successional status in 80 woody species from south Brazil. Journal
of Tropical Ecology 19: 315–324.
New Phytologist (2014) 204: 192–200
www.newphytologist.com
 New
200 Research Phytologist

Table S1 A complete list of data used in the meta-analysis,

Supporting Information
Additional supporting information may be found in the online
version of this article.
Fig. S1 A phylogeny of the taxa used in the meta-analysis, based
on the Angiosperm Phylogeny Group III phylogeny (http://
www.mobot.org/mobot/research/APweb/).
including taxonomic status of taxa and growth forms, grouped by
study identity
Please note: Wiley Blackwell are not responsible for the content
or functionality of any supporting information supplied by the
authors. Any queries (other than missing material) should be
directed to the New Phytologist Central Office.

New Phytologist is an electronic (online-only) journal owned by the New Phytologist Trust, a not-for-profit organization dedicated
to the promotion of plant science, facilitating projects from symposia to free access for our Tansley reviews.

Regular papers, Letters, Research reviews, Rapid reports and both Modelling/Theory and Methods papers are encouraged.
We are committed to rapid processing, from online submission through to publication ‘as ready’ via Early View – our average time
to decision is <25 days. There are no page or colour charges and a PDF version will be provided for each article.

The journal is available online at Wiley Online Library. Visit www.newphytologist.com to search the articles and register for table
of contents email alerts.

If you have any questions, do get in touch with Central Office (np-centraloffice@lancaster.ac.uk) or, if it is more convenient,
our USA Office (np-usaoffice@lancaster.ac.uk)

For submission instructions, subscription and all the latest information visit www.newphytologist.com

New Phytologist (2014) 204: 192–200 Ó 2014 The Authors

www.newphytologist.com New Phytologist Ó 2014 New Phytologist Trust