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Clarication on rust species potentially infecting pea (Pisum sativum L.) crop
and host range of Uromyces pisi (Pers.) Wint
Eleonora Barilli
a,
*
, Ana Moral
a
, Josena C. Sillero
b
, Diego Rubiales
a
a
Institute for Sustainable Agriculture, CSIC, Apdo. 4084, E-14080 Crdoba, Spain
b
IFAPA, Centro Alameda del Obispo, Apdo. 3092, E-14080 Crdoba, Spain
a r t i c l e i n f o
Article history:
Received 4 July 2011
Received in revised form
4 January 2012
Accepted 8 January 2012
Keywords:
Pea rust
Uromyces pisi
Host range
a b s t r a c t
Rust is a serious disease of pea whose casual agent is not always understood. In this paper we studied
reaction of pea accessions to seven rust species infecting closely related legumes, nding that indeed pea
can be infected mainly by Uromyces pisi, followed by Uromyces viciae-fabae. Other rust species like
Uromyces striatus, Uromyces ciceris-arietini, Uromyces anthyllidis and Uromyces vignae can also infect and
reproduce on pea, although in a minor extent. All U. pisi isolates tested were very virulent on pea
accessions, but isolates UpPt-03 and UpKeS-05 (from Palmar de Troya, Spain and Kafr-El-Sheik, Egypt
respectively) were signicantly most infective. In addition to this we studied in detail the host range of
U. pisi by inoculating multiple accessions of various legumes with urediospores of seven isolates of U. pisi
from different geographical origins. Both experiments were performed under controlled conditions.
Based on the evidence presented here the host range of U. pisi is greater than previously recorded,
including genotypes belonging to Cicer arietinum, Vicia articulada, Vicia ervilia and Vicia faba, which were
not mentioned before.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Pea rust is a serious disease of worldwide distribution causing
yield losses in pea (Pisum sativum L.). In years of epidemics, rust
results in affected leaves drying up and detaching fromthe plant, so
beans remain undeveloped and consequently yield losses can be
more than 30% (EPPO, 2010). The pathogen develops in warm,
humid weather and may cause signicant damage when epidemics
start early in the season and when springs are humid and
temperate (Sillero et al., 2006; Emeran et al., 2008).
Most reports refer to Uromyces viciae-fabae (Pers.) J. Schrt (syn.
Uromyces fabae Pers de Bary) (Xue and Warkentin, 2001; Singh
et al., 2004; Vijayalakshmi et al., 2005; Kushwaha et al., 2006) as
the causal agent of pea rust. However, few observations on
morphology or attempts for classication are provided in these
reports. It was recently suggested that Uromyces pisi (Pers.) Wint. is
the principal agent causing pea rust at least in temperate regions
(Barilli et al., 2009a), what is in agreement with earlier observations
by Gumann (1959). Although pea seedling could be infected by
U. viciae-fabae under controlled conditions, pea accessions were
little infected under eld conditions by isolates from Australia, the
Netherlands, Spain or Syria (Barilli et al., 2009a). Little is known of
pathogenicity on pea by other rusts species.
U. pisi is macroscopically identical to U. viciae-fabae in the ure-
dial stage (being urediospores responsible for disease development
under eld conditions causing a multi-cycling infection), but they
can be distinguished by the morphology of telia and infection
structures (Emeran et al., 2005; Sillero et al., 2006) as well as by
molecular tools (Emeran et al., 2008; Barilli et al., 2011). It seems
that U. pisi is less specialised than other Uromyces species
(Gumann, 1959; Emeran et al., 2008), but its host range has not yet
been claried. Knowledge about host range of a biotrophic fungus
(as Uromyces) is of high agronomic and epidemiologic importance.
In fact, one of the constraints showed by the species belonging to
this genus is that several rust species may infect the same host
plants ex. U. viciae-fabae and U. pisi on pea (Barilli et al., 2009b). In
addition, it is possible that a rust fungus may infect a plant species
that previously was thought to be resistant ex. Medicago spp. that
was recently added to Uromyces ciceris-arietini host range
(Stuteville et al., 2010). These features impede a clear pathogen
characterisation and, as consequence, its control.
The purpose of the research reported here were (i) to determine
the host status of pea against other major legume rusts, (ii) to
* Corresponding author. Tel.: 34 957499211; fax: 34 957499252.
E-mail addresses: ebarilli@ias.csic.es, e.nora@libero.it (E. Barilli).
Contents lists available at SciVerse ScienceDirect
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doi:10.1016/j.cropro.2012.01.019
Crop Protection 37 (2012) 65e70
Author's personal copy
further dene the host range of U. pisi and (iii) to compare the
virulence of U. pisi isolates with different geographical origin.
2. Materials and methods
2.1. Experiment 1: studying susceptibility of pea to different
Uromyces spp.
2.1.1. Fungal isolates and pea accessions
Three monopustular isolates of U. pisi from different geographic
areas and a monopustular isolate each of Uromyces anthyllidis
(Grev.) Schrt., U. ciceris-arietini Jacz. in Boyer & Jacz., Uromyces
striatus J. Schrt., U. viciae-fabae (Pers.) J. Schrt. ex Vicia faba,
U. viciae-fabae (Pers.) J. Schrt. ex Vicia sativa and Uromyces vignae
Barclay (Table 1) were used in the experiment. Isolates were
multiplied on highly susceptible cultivars of their different hosts:
pea (P. sativum cv. Messire), fenugreek (Trigonella foenum-graecum
L. landrace from Tunisia), chickpea (Cicer arietinum L. cv. Fardn),
barrel medic (Medicago truncatula Gaertn. cv. Baraka), faba bean (V.
faba L. cv. Baraca), common vetch (V. sativa L. cv. Mezquita) and
cowpea (Vigna unguiculata (L.) Walp. landrace from Egypt).
Accessions (listed in Table 2) of Pisum sativum ssp. abysinicum,
P. sativumssp. arvense, P. sativumssp. elatius, P. sativumssp. jornadii,
P. sativum ssp. sativum, P. sativum ssp. syriacum, P. sativum ssp.
thebaicum, and P. fulvum were used in the study.
2.1.2. Host plant propagation and inoculation
To ensure experiments with a uniformplant development stage,
seeds were germinated during 48 h on wet lter paper in a Petri
dish at 4

C. The Petri dishes where then transferred to 20

C during
5e7 days. Germinated seeds were planted into plastic pots
(6 6 10 cm) lled with a 1:1 mixture of sand and peat in a rust-
free growth chamber. There were three consecutive replications
per fungal isolate, arranged in a complete randomized design. Each
replicate consisted of 3 pots, each pot containing between 3 and 5
plants of each accession depending on their availability. Pea cv.
Messire was included in each replication as a susceptible check.
Inoculation was done on three-week-old plants, by dusting the
plants with Uromyces urediospores (2 mg spores plant
1
) diluted in
pure talc (1:10, w:w) using a spore settling tower. Plants were
incubated for 24 h at 20

C in complete darkness and 100% relative
humidity, then transferred to a growth chamber at 20

C, under
a photoperiod of 14/10 h day/night regime, with 148 mmol m
2
s
1
irradiance at plant canopy, as previously mentioned. Every exper-
imental set of plants per isolate was maintained apart from the
others in distinct growth chambers.
2.1.3. Disease evaluation and statistical analysis
Infection type (IT), and the disease severity (DS) were
observed 15 days after inoculation, when sporulation occurred
profusely in the susceptible plants. IT was assessed using the 0e4
scale of Stackman et al. (1962), where IT 0 no symptoms,
IT ; necrotic ecks, IT 1 minute pustules barely sporulating, IT
2 necrotic halo surrounding small pustules, IT 3 chlorotic
halo and IT 4 well-formed pustules with no associated chlorosis
and necrosis. DS was visually estimated as the percentage of
symptomatic area of the whole plant, this is, including both
percentage of pustules and percentage of chlorotic or necrotic
tissue.
For statistical analysis, DS values were angular transformed
(180/p arcsine [O(%/100)]) and combined from the three exper-
iments to determine average ratings. Comparisons of DS values
among accessions of the same species and between different
isolates were subjected to an ANOVA and mean values were
separated by LSD test at P < 0.01. Statistical analyses were per-
formed using Statistix (version 8.0; Analytical Software, Talla-
hassee, USA).
2.2. Experiment 2: determining the U. pisi host range
2.2.1. Fungal isolates and legume accessions
Seven monopustular isolates of U. pisi from different
geographic areas were used in the study (Table 1). Spores were
preserved in liquid nitrogen and multiplied on pea cv. Messire
before use. Pea plants were grown in pots (6 by 6 by 10 cm) lled
with a 1:1 mixture of sand and peat. When their third or fourth
leaves were fully expanded, inoculation was carried out by
dusting the plants with rust urediospores. Plants were incubated
for 24 h at 20

C in complete darkness and 100% relative
humidity, and then transferred to a growth chamber at 20

C
under a photoperiod at 14 h light and 10 h dark with light
intensity of 148 mmol m
2
s
1
irradiance. After two weeks,
urediospores were recollected by means of a vacuum spore
collection device, dried during 24 h in Silica Gel supplied by
Merck (Darmstadt, Germany) and stored in refrigerators at 80

C
until required for the experiment. Prior to each experiment,
urediospores were removed from storage and heat shocked at
40

C for 5 min. Each isolate was multiplied apart from the others
to avoid contaminations.
Accessions of alfalfa (Medicago sativa L.), barrel medic
(M. truncatula), blister vetch (Vicia ervilia (L.) Willd.), chickpea (C.
arietinumand Cicer reticulatumL.), chickling pea (Lathyrus cicera L.),
common vetch (V. sativa), cowpea (V. unguiculata), faba bean (V.
faba), lentil (Lens culinaris Medik.), narbon bean (Vicia narbonensis
L.), oneower vetch (Vicia articulata Hornem.) and pea (P. sativum)
were used in the study. From 2 to 6 different accessions per species
were tested (Table 3). Pea cv. Messire was used as susceptible
control in each experiment. Seeds were kindly provided from CGN,
Wageningen, Holland (CGN-numbers) and USDA, Washington State
University, USA (PI-numbers).
2.2.2. Host plant propagation and inoculation
To ensure experiment with a uniform plant development
stage, seeds were germinated during 48 h on wet lter paper in
a Petri dish at 4

C, as described in experiment 1. Seeds of V.
ervilia, V. narbonensis and V. unguiculata were previously scaried
by nicking with a razor blade. Replications, experimental design
and inoculation were described in experiment 1.
2.2.3. Disease evaluation and statistical analysis
Infections type (IT) and the disease severity (DS) were observed
as described above. For statistical analysis, data were treated as
Table 1
Codes of reference, geographical origin and specie of the Uromyces isolates used in
growth chamber experiments at Cordoba (Spain).
Fungal
code
Specie Plant specie Collecting site
UaT-01 Uromyces
anthyllidis
T. foenum-graecum Beja, Tunisia
UcaC-01 U. ciceris-arietini C. arietinum Cuenca, Spain
UpCo-01 U. pisi P. sativum Crdoba, Spain
UpKeS-05 U. pisi P. sativum Kafr-El-Sheik, Egypt
UpMo-05 U. pisi P. sativum Morocco, unknown location
UpPt-03 U. pisi P. sativum Palmar de Troya, Spain
UpRa-05 U. pisi P. sativum Rabat, Morocco
UpWi-02 U. pisi P. sativum Winnipeg, Canada
UpZe-04 U. pisi P. sativum

Zidenice, Czech Republic
UsC-01 U. striatus M. sativa Crdoba, Spain
UvfCo-01 U. viciae-fabae V. faba Crdoba, Spain
UvsCo-02 U. viciae-fabae V. sativa Crdoba, Spain
UvC-01 U. vignae V. unguiculata Canada, unknown location
E. Barilli et al. / Crop Protection 37 (2012) 65e70 66
Author's personal copy
explained in experiment 1, using Statistix (version 8.0; Analytical
Software, Tallahassee, USA).
3. Results
3.1. Experiment 1
All Pisum accessions studied displayed a compatible interaction
to the three isolates of U. pisi tested (Table 2) but DS values ranged
from 0 to 45%. Accessions belonging to P. sativum ssp. arvense and
ssp. thebaicumwere the most susceptible to U. pisi isolate UpCo-01,
displaying higher DS values (>32%). Pisum fulvum accessions also
displayed compatible IT but very low DS (<7%) with the exception
of P651. Likewise, U. pisi isolates UpWi-02 and UpPt-03 induced
a compatible response in the whole collection, with DS values
ranging from 3 to 46%. P. sativum ssp. thebaicum and ssp. abyssini-
cum were respectively the most susceptible to each isolate
(Table 2). Considering the whole Pisum collection, no signicant
differences were found between U. pisi isolates, which showed
similar infection values.
U. anthyllidis isolate UaT-01 caused low infection types (IT ; and
2) and reduced disease severity values (DS <5%) in most accessions
studied, causing susceptible reaction in some accessions of
P. sativum ssp. abyssinicum and ssp. elatius (Table 2). Differences
between subspecies were not signicant (P 0.382).
Infection caused by U. ciceris-arietini (UcaC-01) was signicantly
reduced compared to those achieved by pea, faba bean and vetch
rusts (P < 0.05), with 3 accessions being either immune (IT 0) (P27,
Table 2
Macroscopic response of a Pisum spp. collection against inoculation with different Uromyces isolates under controlled conditions. Infection type (IT) and % of disease severity
(DS) are reported. Data followed with different small letters, per column and Pisum subspecies, are signicantly different (LSD P < 0.01). Data followed with different capital
letters compared, per column, averages between Pisum subspecies (LSD P < 0.01).
Code Species Subsp. UpCo-01 UpWi-02 UpPt-03 UaT-01 UcaC-01 UsC-01 UvfCo-01 UvsCo-02 UvC-01
IT
a
DS
b
IT DS IT DS IT DS IT DS IT DS IT DS IT DS IT DS
P1 P. sativum abyssinicum 4 30ab 4 6.7b 4 32.5ab ; e 2 5.0b 4 0.5a 4 26.0bc 4 6.3bc 4 12.0a 4 1.9a
P2 P. sativum abyssinicum 4 34.2a 4 46.0a 4 31.0ab 4 15.0a 4 0.5a 4 41.0a 4 17.0a 4 10.0a 3 0.6a
P7 P. sativum abyssinicum 4 20.2b 4 11.7b 4 34.0a 4 4.4b 4 3.2a 4 33.0ab 4 20.0a 4 7.0a 4 0.3a
P9 P. sativum abyssinicum 4 39.2a 4 41.7a 4 24.0b 2 0.9b 4 0.1a 4 22.0bc 4 4.2c 4 12.0a 4 0.5a
P10 P. sativum abyssinicum 4 22.5b 4 10.0b 4 35.0a 2 0.6b 4 4.6a 4 16.3c 4 12.5ab 4 6.3a 0a
Average 29.2AB 23.2A 31.3A 5.2A 1.7A 27.7A 11.9A 9.5A 0.7A
P13 P. sativum elatius 4 28.3a 4 18.0b 4 27.5a 4 41.0a 4 0.8a 4 17.5a 4 41.0a 4 4.2c 4 1.8a
P14 P. sativum elatius e e 4 26.0a 4 38.8a ; 1a ; 1.0c 4 38.8a 4 10.0b 4 0.6a
P18 P. sativum elatius 4 22.5ab 4 15.0b 4 26.3a 4 3.7b e e 4 3.7b ; 3.0c 4 0.3a
P19 P. sativum elatius 4 12.1b 4 28.0a 4 25.0a 4 9.0b 4 0.2b 4 9.0b 4 9.0b 4 18.0a 4 0.6a
P20 P. sativum elatius e 4 5.0c 4 25.0a ; 2.3b e ; 5.2bc ; 2.3b ; 2.3c 0a
Average 20.9AB 16.5A 25.9AB 18.9A 0.4A 5.1B 18.9A 7.5A 0.6A
P26 P. sativum sativum 4 16.7b 4 30.0a 4 26.6a ; 1.6a 4 0.1a 4 21.7a 4 13.3ab 4 0.5b 0b
P27 P. sativum sativum 4 28.0ab e e ; 4.2a 0a e e ; 3.7a 4 0.5ab
P28 P. sativum sativum 4 45a 4 23.3a 4 25.0a 4 2.6a 4 0.5a 2 12.5a 4 3.6b 0b 3 0.1b
P30 P. sativum sativum 4 19.2b 4 27.0a 4 33.0a e 4 2.7a 4 22.0a 4 18.0a ; 5.0a 4 0.7a
P31 P. sativum sativum e e e ; 1.0a e e 4 10.0ab e 4 0.5ab
Average 27.2AB 26.8A 28.2AB 2.4A 0.8A 18.7AB 11.2A 2.3A 0.4A
P41 P. sativum arvense e 4 26.0b e e e e e e e
P42 P. sativum arvense 4 36.0a e e e e e e e e
P316 P. sativum arvense 4 32.5a 4 40.0a 4 22.0a ; e 3 4.0a 4 1.8a 4 10.0a 4 3.4a 2 7.0a 4 0.4a
P626 P. sativum arvense 4 40.0a 4 28.0b 4 25.0a ; 0.4b ; 0.4a 4 11.3a 4 10.0a 2 3.4a 4 0.7a
P627 P. sativum arvense e 4 21.7b 4 23.0a ; 0.4b 4 0.4a ; 5.3a 4 6.7a 2 5.0a 4 0.4a
Average 36.3A 28.9A 23.3AB 1.6A 0.9A 8.8B 6.7A 5.1A 0.5A
P617 P. sativum thebaicum 4 34.2 4 31.7 4 37.0 ; 2.6 4 0.5 ; 23.0 4 7.0 4 1.6 3 0.5
Average 34.2A 31.7A 37.0A 2.6A 0.5A 23.0AB 7.0A 1.6A 0.5A
P311 P. sativum syriacum 4 14.2a 4 12.0b 4 26.3a ; 1.3a 0b 4 17.5a 4 0.8b 4 7.5a 0b
P665 P. sativum syriacum 4 12.0a 4 35.0a 4 34.0a ; e 2 0.5a 4 5a 4 15.0a 4 28.0a 4 6.3a 4 0.9a
Average 13.1BC 23.5A 30.1AB 0.9A 2.5A 16.2AB 14.4A 6.9A 0.5A
P51 P. sativum jornadii e e e e e e e e e
P621 P. sativum jornadii 4 25.8a 4 31.3a 4 13.8a ; 2.5a 4 7.0a ; e 2 16.7a 4 19.0a ; 7.0a 4 0.4a
P649 P. sativum jornadii 4 6.7a 4 8.3b 4 20.0a ; e 2 3.7a 4 5.5a ; e 2 17.5a e ; 3.7b 4 0.1a
P650 P. sativum jornadii 4 18.7a 4 23.3ab 4 26.7a e 4 2.6a 2 16.0a 4 5.3b ; 13.3a 4 0.5a
Average 17.1BC 20.9A 20.1AB 3.1A 5.0A 16.7AB 12.2A 8.0A 0.3A
P651 P. fulvum 4 22.5a 4 17.5a e e 0b ; 4.0a 4 8.0ab 4 23.0a 4 0.5a
P658 P. fulvum 4 2.6b 4 23.0a 4 20.0a e 4 0.3ab ; 2.0a 4 13.7a 4 2.3b 4 0.5a
P660 P. fulvum 0 0b 4 6.2b 4 3.0b ; e 2 1.0a ; 0.9a ; e 2 3.0a 4 7.0ab ; 0.8a 3 0.5a
P661 P. fulvum 4 1b 4 22.0a 4 16.4a e ; 0.5ab ; e 2 5.0a 4 17.8a 4 3.7b e
P663 P. fulvum 4 7b 4 18.0a 4 21.3a 4 e ; 0.4a 4 0.1b 4 1.0a 4 4.2b 4 24.0a 3 0.4a
Average 6.6C 17.3A 15.2B 0.7A 0.4A 3.0B 10.1A 10.8A 0.5A
LSD 0.009 0.792 0.0085 0.382 0.239 0.003 0.967 0.401 0.997
e Non determined.
a
IT: Infection type following Stackman et al. (1962) scale.
b
DS: nal disease severity (%) measured under controlled conditions.
E. Barilli et al. / Crop Protection 37 (2012) 65e70 67
Author's personal copy
P311 and P651), 4 showing hypersensitive response (IT ;) (P14,
P626, P660 and P661) or displaying a compatible interaction but
very low DS (<7.0%) (Table 2).
U. striatus (UsC-01) was virulent on all P. sativum ssp. abyssini-
cum, ssp. sativum (except on accession P28) and ssp. syriacum
accessions (IT 4, DS >16%) (Table 2). Accessions from P. sativum ssp.
elatius, ssp. arvense and P. fulvum showed either compatible or
incompatible interaction, with signicantly reduced DS values
(P 0.003). All the accessions from P. sativum ssp. thebaicum and
ssp. jornadii showed hypersensitive reaction.
Table 3
List of legume hosts that were inoculated with isolates of U. pisi under controlled conditions, infection type (IT) and % of disease severity (DS) for each species and isolates. Data
followed with different letters, per column and host species, are signicantly different (P < 0.01).
Host specie Code UpCo-01 UpKeS-05 UpMo-05 UpPt-03 UpRa-05 UpWi-02 UpZe-04
IT
a
DS
b
% IT DS% IT DS% IT DS% IT DS% IT DS% IT DS%
Pisum sativum Messire
c
4 47a 4 71b 4 47a 4 80a 4 44a 4 39ab 4 30ab
P. sativum Ballet
c
4 44a 4 49c 4 16b 4 60a 4 15c 4 30b 4 34ab
P. sativum Athos
c
4 31ab 4 22d 4 21b 4 58a 4 27b 4 52a 4 19b
P. sativum PI347347 4 10b 4 80a 4 13b 4 66a 4 22b 4 37ab 4 40a
P. sativum PI343965 4 12b 4 72b 4 17b 4 60a 4 20b 4 29b 4 38ab
Lathyrus cicera BG1043 4 38a 4 30a 4 37a e 4 40a 4 26b 4 41a
L. cicera BG23558 2 30a 2 31a 2 28a e 2 40a 2 37a 2 32a
Vicia articulata BGE001098 e e 4 11a 4 22a 4 11b 4 23a e e 4 12a
V. articulata BGE001149 e e 4 11a 4 18ab 4 11b 4 27a e e 4 5b
V. articulata BGE001169 e e 4 14a 4 13b 4 18a 4 16b e e 4 2b
V. articulata BGE001384 e e 4 10a 4 19ab 4 22a 4 14b e e 4 4b
Lens culinaris L1 1 12 4 5 0 0b e ; 5a 4 4 4 3b
L. culinaris L2 e e e e 2 4a e 3 7a e e ; e 4 15a
Cicer arietinum Fardn
c
; 11ab 4 18a 2 4a 2 3bc 3 15a e e 3 7b
C. arietinum JG62 2(4) 15a 4 13a 4 5a 4 7b 3 11a 4 14ab 3 4b
C. arietinum ILC3279 ; 9b 2 14a 1 3a 4 5bc 2 11a 2 19a e e
C. arietinum WR315 ; 11ab 4 20a 1 7a ; 13a ; e 3 12a 4 25a ; e 3 18a
C. arietinum CA2156 2 12ab 3 15a 4 2a ; 3bc 3 12a 4 17ab 0 0b
C. reticulatum Cr5-10 1 22a 3 12a 2 3a 0 0c e e 4 23a 3 9b
Vicia ervilia PI212333 ; 20ab 2 29ab ; 10b e e ; 35a 2 10b ; 34a
V. ervilia PI220884 ; 12b 1 14c ; 5c e e ; 16bc ; 13ab ; 4b
V. ervilia PI253998 4 33a 4 37a 4 34a 4 21a ; 32ab 4 30a 4 38a
V. ervilia PI284321 ; 21ab ; 22bc ; e 2 6c ; 10b ; 22ab 2 18ab ; 23a
V. ervilia PI518455 ; 21ab ; 17c ; e 2 7bc e e ; 16c 2 14ab ; 24a
Vicia faba Baraca
c
; (2) 9b 2 9b 2 3ab 4 6a 2 6a 2 6a 2 6a
V. faba Brocal
c
2 4bc 2 11b 2 1b ; 1ab 2 2b 2 4a 3 8a
V. faba BPL261 ; e 2 13ab 3 19a 4 2ab 0 0b 3 10a 2 17a 2 8a
V. faba V1273 ; (2) 13ab 2 10b 2 6a 2 5a 2 4a 2 9a 2 16a
V. faba 2N52 ; e 2 11ab 4 (2) 7b 4 2ab ; 1b 2 (4) 6a 2 8a e e
Vicia narbonensis IFVN563 sel2471 ; 22ab ; 28ab ; 3a 0 0b ; 21a 0 0b e e
V. narbonensis IFVN565 sel2473 ; 23a ; 38a ; 2a ; 5a ; 23a ; 16a ; 3b
V. narbonensis IFVN573 sel2487 ; 19ab ; 37a ; 4a ; 7a ; 15a ; 20a ; 18a
V. narbonensis IFVN576 sel2390 ; 8b ; 12c ; 5a 0 0b ; 18a ; 4b e e
V. narbonensis IFVN560 sel2468 ; 6b ; 23bc ; 3a ; 3a ; 18a e e e e
Vicia sativa Mezquita
c
; 10a ; 9a 0 0 e e ; 2a ; 11a ; 1a
V. sativa PI284080 0 0b 0 0b 0 0 0 0 0 0b 0 0b 0 0a
Medicago sativa Baraka
c
0 0 0 0b 0 0 0 0 0 0 0 0 0 0
M. sativa Ampurdan
c
0 0 0 0b 0 0 0 0 0 0 0 0 0 0
M. sativa Arthur
c
0 0 0 0b 0 0 0 0 0 0 0 0 0 0
M. sativa Victoria
c
0 0 ; 4a 0 0 0 0 0 0 0 0 0 0
M. sativa Paraggio
c
0 0 0 0b 0 0 0 0 0 0 0 0 0 0
Medicago truncatula Parabinga
c
0 0 0 0 0 0 0 0 0 0 0 0 0 0
M. truncatula SA1316 0 0 0 0 0 0 0 0 0 0 0 0 0 0
M. truncatula SA1326 0 0 0 0 0 0 0 0 0 0 0 0 0 0
M. truncatula SA9710 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Vigna unguiculata L1 0 0 0 0 e e ; 75 0 0 ; 3a 0 0
V. unguiculata L2 0 0 e e e e e e 0 0 0 0b 0 0
e Non determined.
a
Infection type following Stackman et al. (1962): IT 0 no symptoms, IT ; = necrotic ecks, IT 1 minute pustules barely sporulating, IT 2 necrotic halo surrounding small
pustules, IT 3 chlorotic halo and IT 4 well-formed pustules with no associated chlorosis and necrosis.
b
DS: nal disease severity (%) measured under controlled conditions.
c
Commercial varieties are named.
E. Barilli et al. / Crop Protection 37 (2012) 65e70 68
Author's personal copy
U. viciae-fabae isolate UvfCo-01 (collected on faba bean)
successfully infected most Pisum spp. accessions studied, but
causing lower DS than U. pisi isolates (Table 2). DS values were not
signicantly different within Pisum spp. (P 0.97). P. sativum ssp.
elatius accession P20 was the only displaying hypersensitive
reaction.
U. viciae-fabae isolate UvsCo-02 (collected on vetch) caused
a compatible interaction (IT 4) on all P. sativum ssp. abyssinicum,
ssp. thebaicum and ssp. syriacum accessions but with low severity
(DS < 12%) (Table 2). All P. sativum ssp. arvense and ssp. jornadii
accessions were resistant displaying incompatible IT (; e 2).
P. sativum ssp. sativum, ssp. elatius and P. fulvum showed both
susceptible and hypersensitive reaction, with DS values less than
24%. Generally, infection caused by vetch rust was signicantly
reduced than pea rust disease (P < 0.05).
U. vignae isolate UvC-01 caused a compatible interaction (IT 3
and 4) but with very low infection levels (DS < 1.9%) on all Pisum
spp. tested (Table 2).
3.2. Experiment 2
U. pisi showed, on the whole leguminous plants collection,
signicant difference in pathogenicity depending on the isolate
tested.
All pea (P. sativum) accessions showed a compatible interaction
(IT 4) against all the isolates of U. pisi tested (Table 3), but level of
infection varied greatly (DS from10 to 80%). Both UpPt-03 (DS
58e80%) and UpKeS-04 (DS 22e80%) isolates were especially
virulent (P < 0.01). Remaining isolates gave a similar level of
severity (range average DS 22e38%). P. sativumaccessions PI347347
and PI343965, which were previously selected for their resistance
against UpCo-01 (Barilli et al., 2009b,c), showed the lowest DS
values against isolates UpCo-01, UpRa-05 and UpMo-05.
One of the L. cicera accessions (BG1043) displayed a fully
compatible interaction (IT 4) to all the isolates with high DS,
whereas the other accession (BG235558) displayed incompatible
reaction (IT 2).
All U. pisi isolates tested caused a compatible interaction (IT 4)
on all V. articulata accessions, although gave only low to moderate
DS values (<27.1%). In general, differences among accessions were
signicant (P < 0.01) (Table 3). Isolate UpRa-05 was the most
virulent.
Reaction of L. culinaris accessions varied with the isolate tested,
but always displaying low DS. Accession L1 displayed incompatible
interaction (IT 0e1) to isolates UpCo-01, UpMo-05 and UpRa-05,
whereas it showed a compatible interaction (IT 4) although with
low DS (<5%) to UpKeS-05 and UpWi-02 infection. Accession L2
showed incompatible interaction (IT 2) against isolate UpMo-05
and compatible interaction (IT 3e4) against UpRa-05 and UpZe-
04, despite low DS values. Reaction of C. arietinum and
C. reticulatum accessions varied greatly with the U. pisi isolate
employed, as well as the accession tested (Table 3). These legumi-
nous did not showed, in general, any clear pattern of response
displaying fromno symptoms (IT 0) to compatible interaction (IT 4),
although DS values displayed were low to moderate (<25%).
Isolates UpWi-02 and UpKeS-04 were more virulent while all
accessions were resistant to isolate UpCo-01.
On the contrary, most V. ervilia accessions displayed an incom-
patible interaction (IT ; to 2) to all U. pisi isolates, with the exception
of accession PI253998 that was susceptible to 6 out of the 7 isolates
tested. Most V. faba accessions showed low IT in response to pea
rust isolates, although some accessions displayed a compatible
reaction (IT 3 or 4) against UpKeS-05, UpMo-05 and UpPt-03, but
always with low DS. All V. narbonensis accessions were highly
resistant to all isolates with no sporulation (IT 0) or hypersensitive
reaction (IT ;). However, a substantial amount of necrotic ecks
were formed in some accessions reaching till 38%. Differences were
signicant among accessions and isolates (P < 0.01), being UpKeS-
04 the isolate which caused the highest damage.
All accessions studied of V. sativa, M. sativa, M. truncatula and
V. unguiculata were very resistant to all fungal isolates, with IT
ranging from 0 and ;.
Isolate UpCo-01 was most infective in Pisum and Lathyrus
accessions, showing DS values >35%. Accessions of V. ervilia,
V. articulata, V. narbonensis and Cicer spp., showed moderate
infection levels (DS 10e23%), whereas accessions of V. unguiculata,
V. faba, L. culinaris and V. sativa displayed lower susceptibility to the
pathogen (DS < 8%). M. truncatula and M. sativa were completely
resistant.
4. Discussion
Knowledge of the host range of U. pisi is important to determine
whether it could affect crops other than pea. Furthermore, it is
possible that alternative hosts could provide a means of over-
wintering of the pathogen, providing inoculums to initiate
epidemics in future years. Conversely, pea crops could also be
infected by other rust species.
U. pisi is largely a European pathogen but has also been reported
from Africa, Asia and South America (Erdo gdu et al., 2010). U. pisi is
reported as having a complex life cycle with spermagonia and aecia
developing on Euphorbia spp., and uredinia and telia on some
Fabaceae hosts (Wilson and Henderson, 1966). However, only
limited studies on pea rusts host range have been developed
(Gumann, 1959; Wilson and Henderson, 1966; Conner and Bernier,
1982). Furthermore, most of them are not recent so, as conse-
quence, little is known about its actual virulence.
The reaction of pea against different rust species was studied
here. As expected, isolates from U. pisi successfully infected all the
Pisum spp. tested, being DS values not signicantly different
between isolates UpCo-01, UpWi-02 and UpPt-03. P. fulvum acces-
sion P660 was immune to isolate UpCo-01, as well as showed the
lowest DS values against the others pea rusts. This accession was
identied as an important source of resistance against pea powdery
mildew (Erysiphe pisi DC) and ascochyta blight (Mycosphaerella
pinodes (Berk & Blox) Vesterg) (Fondevilla et al., 2005, 2006) that is
included in our department plant breeding programme. Besides
U. pisi, peas are also infected by U. striatus and, although slightly by
U. viciae-fabae, U. ciceris-arietini, U. anthyllidis and U. vignae.
U. striatus has awidehost rangeanda possible infectiononpeawas
already described (Skinner and Stuteville, 1995). In our study, acces-
sions belonging to P. sativum ssp. abyssinicum, ssp. syriacum, ssp. sat-
ivumand ssp. arvense were the most susceptible to UsC-01 infection.
U. viciae-fabae ex. V. faba and ex. V. sativa were able to infect most
of the pea accession but with a reduced severity compared to U. pisi
isolates. These results conrm previous studies which consider the
pathogen U. pisi, as the principal agent causing pea rust in temperate
regions. In fact, although U. viciae-fabae can infect pea under certain
conditions, it is little adapted to infect pea as high levels of pre-
haustorial resistance mechanisms operated in pea reducing path-
ogen multiplication (Barilli et al., 2009a,b,c). On the contrary, in
tropical andsubtropical regions as India, U. fabae is reportedto be the
pathogen causing rust in peas with yield losses up to 50%
(Vijayalakshmi et al., 2005; Kushwahaet al., 2006, 2007; EPPO, 2010).
Concerning the response caused by U. anthyllidis, U. ciceris-
arietini and U. vignae, data recording showed a great variability on
the species tested, proving the weak specialisation of these species
on pea.
The study on U. pisi host range revealed a great variability
within the leguminous accessions studied, ranging from complete
E. Barilli et al. / Crop Protection 37 (2012) 65e70 69
Author's personal copy
resistance with absence of symptoms to full susceptibility with
compatible interaction and abundant sporulation. All U. pisi isolates
successfully infected all P. sativum accessions, but also all
V. articulata ones. V. faba, L. culinaris, V. ervilia, L. cicera and
C. arietinum accessions showed different degrees of susceptibility
depending on the accession and the isolate tested. Susceptibility of
pea, lentil and chickling pea against U. pisi was previously reported
(Sidenko, 1966), but susceptibility in C. arietinum, V. articulata,
V. ervilia and V. faba is described here for the rst time, expanding
the host range of U. pisi.
Complete resistance was expressed by no symptoms or charac-
teristic symptoms of hypersensitive response (IT ; or IT 2). This
hypersensitive reaction is common in biotrophic pathogeneplant
interactions and was described in cereal and legume plant
response to rust (Tiburzy and Reisener, 1990; Niks and Dekens, 1991;
Sillero and Rubiales, 2002). Beside complete resistance, all the
accessions of V. narbonensis, V. sativa, M. truncatula, M. sativa and
V. unguiculata, showed no symptoms or hypersensitive response
against all the isolates tested, where diversity is greatest.
In terms of pathogenicity, results on peas showed that the local
isolate UpCo-01 was not always the most severe. In fact, disease
severity measured on the primary host plants showed that isolates
UpPt-03 and UpKeS-05 (from Palmar de Troya, Spain and Kafr-El-
Sheik, Egypt respectively) were signicantly more virulent, hence
dangerous if introduced in other elds. The similarity between host
range pattern conrms our previous study (Barilli et al., 2011) con-
cerning the low genetic variability seen on the U. pisi isolates used in
thestudy, despitetheir geographical distance(Canada, CzechRepublic,
Egypt, MoroccoandSpain), as well as thefrequent sexual reproduction
in their alternate hosts Euphorbia spp. (Pfunder and Roy, 2000). In
other rust species, reportedgeographic differentiationof isolates were
attributed not only to geographical barriers restricting movement of
urediospores between the regions (Kolmer and Ordoez, 2007) but
also to differing host selection pressures in the regions (Kolmer and
Liu, 2000; Enjalbert et al., 2005; Mebrate et al., 2006). Furthermore,
the sexual reproduction that is frequent in Euphorbia spp. alternate
hosts (Pfunder and Roy, 2006) it seems that did not contribute to
a wide genetic or pathogenetic distance between isolates.
Chemical control is possible (Emeran et al., 2011) but the use of
host plant resistance is the best mean of rust control (Rubiales et al.,
2011), considering that management practices are not always
effective enough and fungicides are economically and environ-
mentally costly (Stoddard et al., 2010). Previous studies showed
that no complete resistance was found in pea against U. pisi, and
that varieties with partial resistance are not yet commercially
available (Barilli et al., 2009b,c).
Although laboratory infection experiments may overestimate
host status for reasons previously discussed, our data suggest that
numerous plant species can be somehow infected by U. pisi. This
pathogen appeared to be quite aggressive in those species and,
depending on the isolate, could become a serious agricultural
problem. However, further experiments will be required to deter-
mine if U. pisi should be a signicant pathogen on these hosts under
eld conditions.
Acknowledgements
Financial support by the Spanish Projects AGL2008-01239 and
P07-AGR02883 is acknowledged.
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