Weberbauerella 1(12): 1 11 ISSN 2414-8814

THEIMPACTOFRHODODENDRONPONTICUMASAN
INVASIVESPECIESONECOSYSTEMFUNCTIONSOF
LANCASTERUNIVERSITYCAMPUS
Matthew CLARE1
1
Lancaster Environment Centre (LEC), Library Avenue, Lancaster University. Lancaster LA1 4YQ, United King-
dom. E-mail: m.clare@lancaster.ac.uk

ABSTRACT
Rhododendron ponticum is thought of as a Stage V invasive species using the Colautti and
MacIsaac framework. This plant has been sighted on Lancaster University campus, and, as a
result, this paper intends to analyse the effects produced by the growth of this species in the
soil, comparing it to the soil that contains native plants species. A worm count, a pH analysis
and a loss of ignition test to study the organic carbon content were performed, and used a TOC
analyser to study the microbial carbon content. The only experiment that provided statistically
significant results was the worm count, which showed a three-fold decrease in the number of
worms in the soil containing Rhododendron plants. Further analysis is needed to determine the
reason why one of the experiments showed significantly different results, whilst the other tests
showed relatively similar results between the soil types. It is possible that a microbiological
analysis of the soil sampled could have shown that different microbes do exist in the soil con-
taining the Rhododendron and this could have been a potential reason for the similarity in some
of the results obtained.
Keywords: Ericaceae, vermifauna, invasive species, soil ecology, biostatistics.

RESUMEN
ElimpactodeRhododendronponticumcomoespecieinvasoraenlasfuncioneseco
sistmicasdelcampusdelaUniversidaddeLancster:Rhododendron ponticum es con-
siderado una especie invasora de Nivel V, segn el marco de trabajo de Colautti y MacIsaac.
Esta planta ha sido observada en el campus de la Universidad de Lancster y, por tal motivo,
este trabajo pretendi analizar los efectos producidos por el crecimiento de esta especie en el
suelo, comparndolo con suelo que rodea a especies nativas. Se realiz un conteo de gusanos,
anlisis de pH y pruebas de prdida de ignicin para estudiar el contenido de carbono micro-
biano. El nico experimento estadsticamente significativo result ser el conteo de gusanos, el
cual mostr que el suelo que rodea a las plantas de Rhododendron presenta una prdida de dos
tercios de la cantidad total de gusanos. Se propone que son necesarios ms anlisis para deter-
minar la razn de por qu uno de los experimentos dio resultados significativos mientras los
otros arrojaron resultados similares entre los tipos de suelo muestreados. Es posible que un
anlisis microbiolgico de los suelos muestreados demuestre que el suelo donde viven indivi-
duos de Rhododendron tiene una composicin bacteriana diferente a la de otras muestras, lo
que podra dar explicacin a la similitud de los resultados obtenidos.
Palabrasclave: Ericaceae, vermifauna, especies invasoras, ecologa del suelo, bioestadstica.

The author(s), 2016/III/31

 Clare: Rhododendron ponticum as an invasive species

INTRODUCTION
An invasive species is defined as a species that exists outside of its native range and causes
environmental harm (Starr & Harrington, 2004). There have been numerous examples and in-
vasive species that can be found in most environments on earth (Young, et al., 2011). The num-
ber of invasive species appears to be increasing and are a major threat to biodiversity (Alexan-
der, Dick, Weyl, Robinson, & Richardson, 2014). Humans have been responsible for a large
number of these invasions, by both the accidental and the intentional relocating species. Famous
examples include the black rat which was thought to have been transported accidentally by
boats, the house cat which accompanied humans as pets, and the cane toad which was released
into Australia in 1935 to control the population of a native beetle. The effects of an invasive
species can lead to a large change in the structure of the ecosystem and cause a complete change
in the types of species that exist in the habitat.

One of the more damaging invasive species is the earthworm (Lumbricidae) which has had
significant effect on the forest structure of Canada and northern USA. Earthworms aerate soil,
accelerate nutrient cycles and increase water storage. These are vital processes for agriculture;
however, they can be damaging in forest ecosystems (Hale, 2007). Earthworms have been
shown to cause erosion and runoff, decreasing the stability of the land, and to change soil chem-
istry principally by increasing levels of calcium oxalate seven-fold in some regions of North
America (Gorres and Melnichuk, s. d.). This is particularly problematic for low vegetation, such
as shrubs and small plants, which have been removed from many woodlands by the change in
soil make-up and chemistry (Choi, 2012). Whilst the large canopy trees can be relatively unaf-
fected by the presence of earthworms, deer and other herbivores will feed on the saplings of
canopy species after the removal of the herbaceous vegetation (Choi, 2012). In some cases, the
changes in soil chemistry have allowed new invasive plants to become more prevalent in the
forests of North America (Gorres and Melnichuk, s. d.). Therefore, it is important to fully un-
derstand the entire implications of each invasive species.

An example of an invasive species in the UK is Rhododendron ponticum L. Colautti and

MacIsaac (2004) suggested a framework for defining invasive species based on their prevalence
within the ecosystem. On this basis, R. ponticum is likely to be a Stage V invasive species as it
is widespread and numerous reducing environmental stability by interrupting reproduction and
dispersal of native plant species (Colautti and MacIsaac, 2004). This plant species has been
shown to have had negative impacts on the richness of native species in woodland areas, par-
ticularly those containing Quercus petraea (Matt.) Liebl. and Ilex aquifolium L. by inhibiting
native woodland regeneration (Manchester and Bullock, 2000). R. ponticum has also been
shown to negatively impact the vegetation and soil in lowland heath areas (Mitchel, Marrs, Le
Duc, & Auld, 1997).

Lancaster University campus has been identified as a site where individuals of Rhododendron
ponticum are present. It is possible to analyse soil surrounding these introduced plants and soil
surrounding native plant species in order to determine the effects of the invasive plant species
on the soil. Earthworms are an important part of European soil ecology and so any differences
in the distribution of earthworms between the soil types should be recognised. Given the native
earthworms may not be familiar with the effects of the R. ponticum on the surrounding soil, it
is hypothesised that the soil taken from sites near native plant species will, on average, contain
a significantly greater number of earthworms. In the same way that earthworms in the USA are
able to alter soil chemistry, it is reasonable to assume that plants might. R. ponticum may, either
via its own biological functions or a symbiotic relationship, alter the soil pH potentially in order
to improve the suitability of the soil to its own needs. It is predicted that the soil from areas

2
 Clare: Rhododendron ponticum as an invasive species

containing R. ponticum would have a significantly lower pH than soil from near native species.
Finally, an analysis of the organic carbon concentration could explain whether the leaves from
this species are decaying in a similar fashion to native species: its leaves are relatively thick and
waxy and, presumably, would be tougher to digest by microbes. Therefore, it is hypothesised
that the organic carbon concentration of the soil from the sites containing R. ponticum would
be significantly lower than that of the soil containing the native plant species.

MATERIALSANDMETHODS
In order to test the above hypotheses, it was necessary to collect soil samples from Lancaster
University campus. There is an array of species present on the campus and therefore it was
important to know which species were native. Ilex aquifolium was an example of a present
native plant species under which soil was sampled, as well as from under R. ponticum bushes.
The soil samples were taken using a spade obtaining approximately 20 cm3 samples. Further
these large samples, a small soil sample of approximately 100g was collected from top 10 cm
of ground soil was collected from each site.

The large soil samples were used to obtain a worm count. The smaller soil samples were used
to study soil pH, organic carbon content, and microbial biomass. pH was measured using a 5 g
sample of soil to which 25 mL of water was added. The sample was mixed regularly for 20
minutes and then a pH-meter was used. The organic matter content was measured by the loss
on ignition, but the sample was at a much higher temperature in the furnace. Because microbes
are closed systems, the carbon trapped inside is not measured in the same manner as the organic
matter content. Therefore, a 5 g sample of each soil was taken and placed in the desiccator along
with a beaker of amylene-stabilised chloroform and some boiling chips. A water vacuum pump
was used to remove all air from the desiccator and the samples were left in the desiccator at 25
o
C for 24 hours, causing the microbes present to burst. This then transferred to an extraction
bottle and 25 mL, 0.5 M K2SO4 was added and placed on the shaker for 30 minutes. A 5 g
sample of the original soil was taken, transferred into a separate bottle and treated similarly.
After being shaken, the liquid is filtered into a sterile tube for analysis by the TOC analyser.
The difference between the fumigated and non-fumigated sample shows the carbon stored in
microbes. Statistical analyses were performed to show significance and, given the non-para-
metric nature of the data, the Mann-Whitney-Wilcoxon analysis was used to compare the data.

RESULTS
Of the performed experiments, only the worm count was statistically significant. The compari-
son between the statistical descriptive results of every experiment can be seen in Tables A, B,
C and D.

The mean number of worms in the soil from around native plant species is greater than that
from under Rhododendron bushes (Fig A). Table E shows the results of the Mann-Whitney-
Wilcoxon Test for statistical significance in the distribution of worms between the two soil
types. The null hypothesis used stated that there is no significant difference in the distribution
of worms between the soil sampled from soil under native plant species and from the soil ob-
tained from under Rhododendron plants. The test assumes that the sample groups are independ-
ent from each other and that the results are ordinal. Both assumptions are met by this data. The
significance calculated by the Mann-Whitney-Wilcoxon Test shows a significance value of
0.024. Therefore, the null hypothesis can be rejected with 95% confidence and it can be stated
that the differences between the distributions of worms across the two sample types are statis-
tically significant.

3
 Clare: Rhododendron ponticum as an invasive species

Number of worms in soil containing Number of worms in soil containing

native plant species rhododendron
Mean 15.5833 Mean 4.5833
Standard Error 4.4525 Standard Error 1.6212
Median 9 Median 1.5
Mode 18 Mode 0
Standard Deviation 15.4241 Standard Deviation 5.6159
Sample Variance 237.9015 Sample Variance 31.5379
Kurtosis 2.6228 Kurtosis -0.4933
Skewness 1.5928 Skewness 0.8965
Range 53 Range 16
Minimum 1 Minimum 0
Maximum 54 Maximum 16
Sum 187 Sum 55
Count 12 Count 12
Confidence Level Confidence Level
9.8 3.5681
(95.0%) (95.0%)
Table A Descriptive statistical analysis of the results from the worm count.

pH of soil containing native plant spe-

pH of soil containing rhododendron
cies
Mean 6.3417 Mean 5.5083
Standard Error 0.4256 Standard Error 0.5082
Median 6.65 Median 4.8
Mode 5.3 Mode 4.1
Standard Deviation 1.4743 Standard Deviation 1.7604
Sample Variance 2.1736 Sample Variance 3.0990
Kurtosis -0.7161 Kurtosis -0.8153
Skewness -0.4956 Skewness 0.7805
Range 4.7 Range 5
Minimum 3.6 Minimum 3.8
Maximum 8.3 Maximum 8.8
Sum 76.1 Sum 66.1
Count 12 Count 12
Confidence Level Confidence Level
0.9367 1.1185
(95.0%) (95.0%)
Table B Descriptive statistical analysis of the results from the study of pH.

4
 Clare: Rhododendron ponticum as an invasive species

Organic matter (%) in soil containing native Organic matter (%) in soil containing
plant species rhododendron

Mean 62.0255 Mean 66.6044

Standard Error 4.9997 Standard Error 7.5330
Median 60.3528 Median 63.6608
Standard Deviation 17.3196 Standard Deviation 26.0949
Sample Variance 299.9668 Sample Variance 680.9445
Kurtosis 1.5861 Kurtosis -1.5110
Skewness 0.4226 Skewness -0.0325
Range 69.9622 Range 74.3291
Minimum 29.7119 Minimum 25.1205
Maximum 99.6741 Maximum 99.4495
Sum 744.3062 Sum 799.2529
Count 12 Count 12
Confidence Level Confidence Level
11.0043 16.5799
(95.0%) (95.0%)
Table C Descriptive statistical analysis of the results from the loss of ignition study for organic matter

Microbial carbon concentration (g/g) Microbial carbon concentration (g/g)

in soil containing native plant species in soil containing rhododendron

Mean 764.3719 Mean 1039.4489

Standard Error 147.5978 Standard Error 240.5207
Median 698.0386 Median 764.5212
Standard Deviation 511.2937 Standard Deviation 833.1882
Sample Variance 261421.2336 Sample Variance 694202.5080
Kurtosis 1.7326 Kurtosis 2.8930
Skewness 1.1375 Skewness 1.5634
Range 1841.2591 Range 2982.3913
Minimum 137.1343 Minimum 164.2768
Maximum 1978.3934 Maximum 3146.6680
Sum 9172.4626 Sum 12473.3866
Count 12 Count 12
Confidence Level Confidence Level
324.8605 529.3825
(95.0%) (95.0%)
Table D Descriptive statistical analysis of the results for microbial carbon content from the TOC analyser.

5
 Clare: Rhododendron ponticum as an invasive species

Fig. A Frequency histogram generated by the Mann-Whitney-Wilcoxon Analysis to distinguish significance in

the distribution of worms between the two soil types. The mean number of worms in the soil from around native
plant species (labelled N) is greater than that from under Rhododendron bushes (labelled I). Each bin represents
five worms.

Table E Results of the Mann-Whitney-Wilcoxon Test for statistical significance in the distribution of worms
between the two soil types (n=12 for both samples).

The soils from both types of location cover a broad range of similar pH values (Fig. B). Alt-
hough the graph does indicate that there are were more sites with a pH under 5.0 in the soil
obtained from under Rhododendron plants, whilst the majority of samples with a pH greater
than 7.0 were obtained from soil surrounding native plant species. Table F shows the results of
the Mann-Whitney-Wilcoxon Test for significant differences in the pH of the soil types. The
null hypothesis used stated that there is no significant difference in the pH of the two soil types
sampled, i.e. soil from under native plant species and from under Rhododendron plants. The
test assumes that the sample groups are independent from each other and that the results are
ordinal. Both assumptions are met by this data. The significance calculated by the Mann-Whit-

6
 Clare: Rhododendron ponticum as an invasive species

ney-Wilcoxon Test shows a significance value of 0.219. Therefore, the null hypothesis is ac-
cepted and it can be assumed that the differences in pH of the different soils are not statistically
significant.

Fig. B Frequency histogram generated by the Mann-Whitney-Wilcoxon Analysis for the pH of soil from under
native plant species (labelled N) and the pH of soil from under Rhododendron plants (labelled I). Each bin repre-
sents 0.5.

Table F Results of the Mann-Whitney-Wilcoxon Test for significant differences in the pH of the soil types
(n=12 for both samples).

The average distribution of organic matter content is reasonably similar between the two soil
types, but does show a negative skew in the distribution of the organic matter content of the
soil collected from soil under Rhododendron plants (Fig. C). The results of the Mann-Whitney-
Wilcoxon for the experiment are shown in Table G. The null hypothesis used stated that there
is no significant difference in the organic matter of the two soil types sampled, i.e. soil from
under native plant species and from under Rhododendron plants. The test assumes that the sam-
ple groups are independent from each other and that the results are ordinal. Both assumptions
are met by this data. The significance calculated by the Mann-Whitney-Wilcoxon Test shows
a significance value of 0.887. Therefore, the null hypothesis is accepted and it can be assumed

7
 Clare: Rhododendron ponticum as an invasive species

that the differences between the organic matter content in the two soil types are not of statistical
significance.

Fig. C Frequency histogram generated by the Mann-Whitney-Wilcoxon Analysis of the organic matter content
of soil from under native plant species (labelled N) and from under Rhododendron plants (labelled I). Each bin
represents 12.5% organic matter.

Table G Results of the Mann-Whitney-Wilcoxon Test for significant differences in the organic matter content in
the two soil types (n=12 for both samples).

Finally, the microbial carbon concentration of soil from under native plant species and from
under Rhododendron plants appear to be similar (Fig. D). The graph shows similar distributions
between the two soil types, although one soil sample from under the Rhododendron plants does
have a higher microbial carbon concentration than any of the samples from the soil from around
native plant species, which appear to be clustered about a lower microbial carbon concentration.
In Table H, the results of the Mann-Whitney-Wilcoxon Test for significant differences in the
Microbial Carbon Concentration between the two soil types are shown. The null hypothesis
used stated that there is no significant difference in the Microbial Carbon Content of the two

8
 Clare: Rhododendron ponticum as an invasive species

soil types sampled, i.e. soil from under native plant species and from under Rhododendron
plants. The test assumes that the sample groups are independent from each other and that the
results are ordinal. Both assumptions are met by this data. The significance calculated by the
Mann-Whitney-Wilcoxon Test shows a significance value of 0.219. Therefore, the null hypoth-
esis is accepted and it can be assumed that the differences in pH of the different soils are not
statistically significant.

Figure D Frequency histogram generated by the Mann-Whitney-Wilcoxon Analysis of the microbial carbon
concentration of soil from under native plant species (labelled N) and from under Rhododendron plants (labelled
I). Each bin represents 12.5% organic matter.

Table H Results of the Mann-Whitney-Wilcoxon Test for significant differences in the microbial carbon con-
centration between the two soil types (n=12 for both samples).

DISCUSSIONANDCONCLUSIONS
The analysis has shown that the distribution of earthworms was significantly different (to a 95%
confidence limit) between the two soil types, with the soil containing native plant species hav-
ing a considerably higher number of worms on average. The analysis of pH showed that, alt-
hough the average pH was lower in soil containing Rhododendron ponticum, the difference is

9
 Clare: Rhododendron ponticum as an invasive species

not statistically significant. Delp (1987) suggested that Rhodondendron can need a soil pH of
between 5.5 and 6.5 for optimum growth. This correlated with the average pH of the soil con-
taining Rhododendron bushes, however there was a relatively large spread of pH in both soil
types. R. ponticum is thought to have a pH lowering effect (Stevens, 2015). This is not obvious
from the results and further testing would be needed.

The average organic content was shown to be 62% 5% (1.S.E.) in the soil containing native
plants and 67% 7% (1.S.E.) in the soil containing Rhododendron ponticum. The microbial
carbon content was shown to be roughly 800 g/g in soil containing native plants and over
1000g/g in soil containing R. ponticum; however, due to the large standard error of data from
the soil containing R. ponticum, the differences in the data sets were deemed statistically insig-
nificant by the Mann-Witney-Wilcoxon analysis.

Only the worm count provided statistically significant data with the number of earthworms in
the soil containing native plant species being considerably higher, as predicted. This is likely
because the Rhododendron ponticum does not provide a familiar environment for native earth-
worm species. There was limited evidence for a significantly lowered pH and therefore it has
not been possible to say that the R. ponticum has had any effect on the pH of the soil. The
carbon contents of the soil types, both free carbon and microbial carbon, were similar and so
the difference in the leaves has not resulted in a decrease in carbon recycling as was expected.
This could have been because there were significant numbers of native plant species surround-
ing the Rhododendron bushes which provided the majority of the leaf litter, or this could have
resulted via the promotion of different microbial species by R. ponticum. Further analysis is
required and should account for this.

ACKNOWLEDGMENTS
I would like to thank Dr. Carly Stevens, Dr. Annette Ryan, and Isabel Rogers of the Lancaster
Environment Centre for help using the TOC analyser, the fumigation equipment, and for in-
struction and aid during the experimental phase. I would also like to thank Dr. Ruth Mitchell of
the James Hudson Institute for conversations regarding the effects of Rhododendron ponticum
on soils in specific environments.

REFERENCES
Alexander, M. E., Dick, J. T., Weyl, O. L., Robinson, T. B., & Richardson, D. M. (2014). Ex-
isting and emerging high impact invasive species are characterized by higher func-
tional responses than natives. Biology Letters, 10, 20130946.
Choi, A. (2012). Invasive Earthworm (Oligochaeta: Lumbricidae) Populations in varying Veg-
etation Types on a Landscape- and Local-scale. Toronto: University of Toronto.
Colautti, R. I., & MacIsaac, H. J. (2004). A neutral terminology to define invasive species.
Diversity and Distributions, 10, 135-141.
Delp, W. E. (1987). Soil pH and Rhododendrons. Journal American Rhododendron Society,
41(4), 225.
Gorres, J., & Melnichuk, R. (s. d.). Invasive Earthworms in Vermont. Retrieved from Vermont
Invasives: http://www.vtinvasives.org/sites/default/files/gorres_invasiveearth-
worms_vermont_0.pdf
Hale, C. (2007). Earthworms of the Great Lakes. Duluth: Kollath-Stensaas Publishing.
Manchester, S. J., & Bullock, J. S. (2001). The impacts of non-native species on UK biodiver-
sity and the effectiveness of control. Journal of Applied Ecology, 37(5), 845-864.

10
 Clare: Rhododendron ponticum as an invasive species

Mitchel, R. J., Marrs, R. H., Le Duc, M. G., & Auld, M. H. (1997). A study of succession on
lowland heaths in Dorset, southern England: changes in vegetation and soil chemical
properties. Journal of Applied Ecology, 34(6), 1426-1444.
Starr, L., & Harrington, E. (Eds.). (2004). National Strategy and Implementation Plan. Wash-
ington, DC: United States Department of Agriculture.
Stevens, P. (2015). Asterid I. Retrieved August 6, 2015, from The Angiosperm Phylogeny Web-
site: http://www.mobot.org/mobot/research/apweb/orders/gentianalesweb.htm#Euas-
teridI
Young, S., Corbin, J., Eviner, V., Dionigi, C., Allen, C., Brooks, C., & Williams, L. (2011).
Ecosystem services the significance of contributions by invasive plant species. Re-
trieved December 20, 2015, from North Central Regional Center for Rural Develop-
ment: http://ncrcrd.msu.edu/uploads/files/133/Ecosystems%20webinar%201-19-
11.pdf

11

The author has requested enhancement of the downloaded file. All in-text references underlined in blue are linked to publications on ResearchGate.