Response of vetches (Vicia spp.) to specialized forms of Uromyces vicia-fabae and to Uromyces pisi

Crop Protection 46 (2013) 38e43

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Response of vetches (Vicia spp.) to specialized forms of Uromyces vicia-fabae and to Uromyces pisi D. Rubiales a, *, J.C. Sillero b, A.A. Emeran c a

Institute of Sustainable Agriculture, CSIC, Apdo. 4084, 14080 Córdoba, Spain IFAPA, Centro Alameda del Obispo, Apdo. 3092, 14080 Córdoba, Spain c Faculty of Agriculture, Kafr El-Sheikh University, 33516 Kafr El-Sheikh, Egypt b

a r t i c l e i n f o

a b s t r a c t s

Article history: Received 18 August 2012 Received in revised form 5 December 2012 Accepted 16 December 2012

The genus Vicia includes many species of agricultural interest, such as faba bean and the various vetches that may be infected by rust, with little understanding of the specificity of these interactions. This work contributes to the clarification of host range of Uromyces viciae-fabae and Uromyces pisi, and confirms host specialization within U. viciae-fabae. The differential response of Vicia faba, Vicia sativa and Lens culinaris checks confirms the existence of specialized isolates of U. viciae-fabae. Host range of these host specialized forms of U. viciae-fabae was not so clear cut when we test other species of Vicia. The most specialized form was U. viciae-fabae ex V. faba that was able to infect profusely only faba bean (V. faba), all other Vicia species being highly resistant. Conversely, susceptibility against U. viciae-fabae ex V. sativa was very common in Vicia spp., particularly in those belonging to the subgenus Vicia, although not in V. faba. Still, susceptibility could be identified in a number of accessions of section Cracca such as Vicia cracca, Vicia monantha, Vicia sicula or Vicia villosa. Most Vicia accessions were highly resistant to U. viciae-fabae ex L. culinaris, but a number of accessions could be severely rusted, particularly in species of the subgenus Vicilla but also in some of section Peregrinae of subgenus Vicia. Similarly, most Vicia accessions were highly resistant to U. pisi but a number of accessions could be severely rusted, particularly in species of the subgenus Vicilla and in sections Hypechusa and Peregrinae of subgenus Vicia. This work also identified sources of resistance to the various rusts that can be used in rust resistance breeding. In spite of the susceptibility against various rusts, interesting levels of resistance were identified in crops and related species being based either on hypersensitive response or on reduced severity in spite of a compatible interaction making resistance breeding feasible. Ó 2012 Elsevier Ltd. All rights reserved.

Keywords: Rust Resistance Vetch Uromyces Vicia

1. Introduction The genus Vicia includes many species of agricultural interest as grain or forage crops, such as faba bean (Vicia faba L.) and vetches, of which the most important are common vetch (Vicia sativa L.) and hairy vetch (Vicia villosa Roth). Many other species such as Vicia ervilia Willd., Vicia narbonensis L., Vicia benghalensis L and Viciaarticulata Hornem are also cultivated although at less extent (Hanelt and Metting, 1989). Both faba bean and vetches can be affected by rust disease incited by Uromyces viciae-fabae (Pers.) J. Schröt (syn. U. fabae Pers de Bary) (Sillero et al., 2006). Chemical control of rust is possible (Emeran et al., 2011), but the use of host plant resistance is the most desired mean of rust control (Rubiales et al., 2011). However,

* Corresponding author. Tel.: þ34 957499215; fax: þ34 957499252. E-mail addresses: [email protected], [email protected] (D. Rubiales). 0261-2194/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.cropro.2012.12.011

little resistance has been described so far in most Vicia species apart from V. faba (Sillero et al., 2000, 2006, 2010). Both for epidemiological and for inheritance studies, more precise knowledge on rust identity and diversity is needed. U. viciae-fabae sensu lato is in fact a species complex with host specialized isolates (Cummins, 1987). Emeran et al. (2005) reported morphological differences in the infection structures of different U. viciae-fabae isolates and suggested subdividing the species into populations with differential pathogenicity against V. faba, V. sativa and Lens culinaris L. This specialization was further supported by the use of RAPD (Emeran et al., 2008) and ITS (Barilli et al., 2011) markers. However, little is known of host range of these rusts in other Vicia species. The purpose of the present study was to characterize the response of a wide range of Vicia species against the above mentioned U. vicia-fabae host specialized isolates and against the close Uromycespisi, in an attempt to clarify the identity of the rusts damaging vetches and to identify sources of resistance.

D. Rubiales et al. / Crop Protection 46 (2013) 38e43

2. Materials and methods A single spore isolate each of U. viciae-fabae ex V. faba, ex L. culinaris and ex V. sativa and of U. pisi were used. Spores stored in liquid nitrogen were multiplied before use on seedlings of the respective host species (Table 1) in isolation in separate growth chambers with filtered ventilation. Seedlings of V. faba (cv. Baraca), L. culinaris (cv. Eston), V. sativa (cv. Mezquita), and Pisum sativum L. (cv. Messire) together with 125 germplasm accessions of Vicia spp. (Table 2) were inoculated in the growth chamber with the above mentioned isolates. Accessions were kindly provided by either IPK, Germany; USDA, USA; Department of Botany of the University of Southampton, UK; or IFAPA, Spain. For each rust test, seed of each accession were planted in three pots (125 ml) with two seedlings per pot. Seedlings of 21day old were inoculated by dusting the plants with a mixture of freshly collected urediniospores in pure talc (1:10). Inoculum concentration was adjusted to 4 mg of urediniospores per seedling. After inoculation, seedlings were incubated at 100% relative humidity at 20  C in darkness for 24 h; and then, they were transferred to a growth chamber at 20  C under a 14 h light: 10 h dark photoperiod with light intensity of 148 mmol/m2/s at the leaf canopy. Twelve days after inoculation infection type (IT) and disease severity (DS) were assessed. IT was recorded following the scale of Stackman et al. (1962) where IT 0 ¼ no symptoms, IT; ¼ necrotic flecks, IT 1 ¼ minute pustules barely sporulating, IT 2 ¼ necrotic halo surrounding small pustules, IT 3 ¼ chlorotic halo and IT 4 ¼ well-formed pustules with no associated chlorosis or necrosis. ITs 0e2 are indicative of resistance, whereas IT 3e4 are of a compatible interaction. Still, within compatible interactions, rate reducing resistance has been reported (Rubiales and Sillero, 2003). DS was recorded as a visual estimation of the percentage of leaf area covered by rust pustules. 3. Results Cross inoculation of the various specialized rusts on their respective hosts confirmed expectations (Emeran et al., 2005), with V. faba cv. Baraca being infected only by U. viciae-fabae ex V. faba; V. sativa cv. Mezquita only by U. viciae-fabae ex V. sativa; L. culinaris cv. Eston only by U. viciae-fabae ex L. culinaris; and P. sativum cv. Messire infected by all tested rusts, but to a major extent by U. pisi and U. viciae-fabae ex L. culinaris. All Vicia accessions, except the V. faba, check were resistant to U. viciae-fabae ex V. faba, showing most of them complete absence of symptoms (IT 0) or up the most only restricted rust development either with reduced IT (accessions 157 of Vicia bithynica, 168, 169, 173, 175 and 176 V. narbonensis, 236 of Vicia pyrenaica, 152 and 155 of V. benghalensis, 35 of Vicia palaestina, 79 of V. ervilia and 147 of V. articulata) or with reduced DS (<15%) in spite of compatible

Table 1 Isolates of Uromyces spp. used to determine their host range on Vicia spp. and species of related genus. Fungal isolate

Species

Geographic origin

Mulitplied on

UvfCo-01

U. viciae-fabae ex V. faba U. viciae-fabae ex L. culinaris U. viciae-fabae ex V. sativa U. pisi

Córdoba, Spain

V. faba cv. Baraca Lens culinaris cv. Eston V. sativa cv. Mezquita Pisum sativum cv. Messire

UvlM-01 UvsCo-02 UpCo-01

Morocco Córdoba, Spain Córdoba, Spain

39

interaction (IT 3) (accessions 240 of Vicia johannis, 250 of Vicia galilea, 29, 170 and 178 of V. narbonensis, 92 of V. sativa, 30 of Vicia hyrcanica, 156 of V. benghalensis, 61 and 206 of Vicia monantha and 272 of V. palaestina). Most Vicia accessions were highly resistant to U. viciae-fabae ex L. culinaris, either with complete absence of symptoms (IT 0) or with restricted rust development either with reduced IT (accession 108 of Vicia lutea, 16 and 132 of V. villosa, and 52, 56, 216 and 217 of Vicia hirsuta). However, a number of accessions could be severely rusted with high IT and DS, particularly in species of the subgenus Vicilla such as V. benghalensis (average DS 12.5%, range 0e60%), V. monantha (average 47.5%, range 35e60%), V. palaestina (average 27.5%, range 25e30%), V. ervilia (40%) and V. articulata (average 20%, range 10e30%), but also in some of subgenus Vicia such in accession of Vicia grandiflora (20%), V. pyrenaica (average 32.5%, range 20e 45%), accession 30 of V. hyrcanica (60%) or Vicia michauxii (average 25%, range 15e40%). Similarly, most Vicia accessions were highly resistant against U. pisi, either with complete absence of symptoms (IT 0) or with only restricted rust development either with reduced IT (accession 168, 169 and 173 of V. narbonensis, 12 and 28 of V. lutea, 75 of V. benghalensis, 35 of V. palaestina, 16 and 134 of V. villosa and 217 of V. hirsuta) or with reduced DS (<10%) in spite of high IT (253 of Vicia hybrida, 30 of V. hyrcanica, 34 of Vicia pannonica, 220 and 257 of V. michauxii, 155 of V. benghalensis and 52 of V. hirsuta) but a number of accessions could be severely rusted with high IT and DS, particularly in species of the subgenus Vicilla such as V. benghalensis, V. monantha, V. palaestina, V. villosa, V. ervilia, Vicia articulata and V. hirsuta, but also in some of subgenus Vicia such in accession of V. bithynica (50% DS), V. grandiflora (40%), V. pyrenaica (60%), accession 36 of V. pannonica (60%), accession 267 of V. michauxii (25%) or Vicia peregrina (average 30%, range 0e60%). Conversely, susceptibility against U. viciae-fabae ex V. sativa was very common in Vicia spp., particularly in those belonging to the subgenus Vicia, with about half of the accessions being highly susceptible, with an average DS of 26%. Species of subgenus Vicia in which all accessions were resistant to the other isolates, showed accessions susceptible to U. viciae-fabae ex V. sativa such as V. johannis (25% DS), V. galilea (20%), V. narbonensis (average 19.4%, range 5e40%), V. sativa (average 26.7%, range 0e50%), Vicia sepium (40%), Vicia cordata (average 18.7%, range 5e30%), and Vicia striata (20%). Other species of the subgenus Vicia in which some accessions could be infected by other isolates also had some accessions susceptible to U. viciae-fabae ex V. sativa, such as V. pyrenaica, V. hyrcanica, V. hybrida, V. michauxii or V. peregrina. In spite of the common susceptibility to this rust in the subgenus Vicia, interesting levels of resistance were identified in most species. Hypersensitive resistance (low IT) was identified in accession 167 and 176 of V. narbonensis, 39 of V. sativa, 39 and 24 of V. sativa, 162 of V. cordata, or 97 of Vicia melanops. Also resistance based on a reduced DS in spite of a compatible interaction was identified in a number of accessions, such as accessions 29, 169 and 175 of V. narbonensis, 245 of Vicia serratifolia, 141 and 144 of Vicia angustifolia, 23 and 95 of V. sativa, 58 and 253 V. hybrida, 165 of V. cordata, or 105 and 106 of V. peregrina. Conversely, most accessions of species of the subgenus Vicilla were highly resistant to U. viciae-fabae ex V. sativa. Still, susceptibility could be identified in a number of accessions of section Cracca such as Vicia cracca (accession 237, DS 25%), V. monantha (average 15.3%, range 1e25%), Vicia sicula (25%) or V. villosa (average 7.9%, range 0e30%). 4. Discussion The genus Vicia is a member of the legume tribe Vicieae of the Papilionoideae along with Lathyrus, Lens, Pisum and Vavilovia. The

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D. Rubiales et al. / Crop Protection 46 (2013) 38e43

Table 2 Response of Vicia spp. accessions to isolates of Uromyces spp. Accession

Donor’s code

Baraca Mezquita Eston Messire 157 240 250 29 168

VIC793/77 NAR 96/86 NAR 137/84 PI29 4301 NAR 129/79

169

NAR 19/75

167 175 178 170 173 176

NAR NAR NAR NAR NAR NAR

245

NAR124/76

39 40 43 82 83 85 86 18 19 20 21 27 92 94 95 96 89 90 41 266 38 42 141 142 144 23 24 25 26 221

PI 173917 PI 173824 PI170474 VIC942/83 VIC 403/79 VIC 573/80 VIC 105/77 PI 284080 PI 284083 PI284085 PI284402 PI183723 VIC 882/80 VIC 354/75 VIC 151/80 VIC 671/74 VIC 838/78 VIC 297/74 PI 393825 VIC 724/84 PI 393867 PI 206394 VIC 719/74 VIC 682/74 VIC579/74 PI207939 PI108273 PI221965 PI244330 VIC 848/79

294 162 163 164 165

93 e Vse4 VIC469/77 VIC470/75 VIC700/74 VIC813/80

282 222 236

26 e VGd3 VIC 48/85 VIC69/86

297 10 30

100 e Vst1 PI 268321 PI 250797

8 57 58

PI 533742 PI 289482 PI 294292

146/84 138/84 131/87 33/77 2/84 43/80

Species

V. faba L. V. sativa L. L. culinaris L. P. sativum L. V. bithynica L. V. johannis var. procumbens H. Schäfer V. galilaea Plitmann & Zohary V. narbonensis L. V. narbonensis var. aegyptiaca Korn. ex Asc. & S. V. narbonensis var. aegyptiaca Korn. ex Asc. & S. V. narbonensis var. affinis Korn. ex Asc. & S. V. narbonensis var. affinis Korn. ex Asc. & S. V. narbonensis var. affinis Korn. ex Asc. & S. V. narbonensis var. narbonensis V. narbonensis var. narbonensis V. narbonensis var. salmonea (Mout.) H. Schäfer Average V. serratifolia Jacq. Average section Narbonensis V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. V. sativa L. var. cosentini (Guss.) Arcang. V. sativa L. var. cosentini (Guss.) Arcang. V. sativa L. var. cosentini (Guss.) Arcang. V. sativa L. var. cosentini (Guss.) Arcang. V. sativa L. var. platysperma V. sativa L. var. platysperma V. sativa L. var. amphicarpa (L.) Boiss. V. sativa L. var. amphicarpa (L.) Boiss. V. sativa L. var. cordata (Hoppe) Arcang. V. sativa L. ssp. nigra (L.) Ehrh. V. sativa L. ssp. nigra (L.) Ehrh. V. sativa L. ssp. segetalis (Thuill.) Arcang. V. sativa L. ssp. segetalis(Thuill.) Arcang. V. sativa L. ssp. nigra (L.) Ehrh. V. sativa L. ssp. nigra (L.) Ehrh. V. sativa L. ssp. nigra (L.) Ehrh. V. sativa L. ssp. nigra (L.) Ehrh. V. sativa L. ssp. macrocarpa (Moris) Arcang. Average V. sepium L. V. cordata Hoppe in Sturm V. cordata Hoppe in Sturm V. cordata Hoppe in Sturm V. cordata Hoppe in Sturm Average V. grandiflora Scop. V. pyrenaica Pourr. V. pyrenaica Pourr. Average Average section Vicia V. striata M. Bieb. V. hyrcanica Fisch. & C. A. Mey. V. hyrcanica Fisch. & C. A. Mey. Average V. hybrida L. V. hybrida L. V. hybrida L.

Subgenus/section

U. viciae-fabae ex V. faba

U. viciae-fabae ex L. culinaris

U. pisi

U. viciae-fabae ex V. sativa

ITa

DSa

IT

DS

IT

DS

IT

DS

Vicia/Bithynicae Vicia/Narbonensis Vicia/Narbonensis Vicia/Narbonensis Vicia/Narbonensis

4 0 ; 4 ; 4 3 3 ;

30 0 0 5 0 1 10 10 0

0 0 4 4 0 nd nd 0 0

0 0 35 20 0 nd nd 0 0

2 0 3 4 4 nd nd 0 ;

1 0 5 30 50 nd nd 0 0

0 4 1 3 0 4 4 4 3

0 20 1 20 0 25 20 15 20

Vicia/Narbonensis

;

0

0

0

1

5

3þ

10

Vicia/Narbonensis Vicia/Narbonensis Vicia/Narbonensis Vicia/Narbonensis Vicia/Narbonensis Vicia/Narbonensis

0 2þ 4 3 1 ;

0 5 1 5 5 0

nd 0 nd 0 0 nd

nd 0 nd 0 0 nd

nd 0 nd 0 1 nd

nd 0 nd 0 5 nd

1þ 4 4 4 4 1þ

5 15 40 25 25 10

Vicia/Narbonensis

nd

Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia

nd 0 0 0 0 0 0 0 0 0 0 0 4 0 0 nd 0 0 0 0 0 0 nd 0 0 nd 0 0 0 0

Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia Vicia/Vicia

0 0 0 0 0

Vicia/Vicia Vicia/Vicia Vicia/Vicia

0 0 2þ

Vicia/Hypechusa Vicia/Hypechusa Vicia/Hypechusa

0 0 4

Vicia/Hypechusa Vicia/Hypechusa Vicia/Hypechusa

0 nd nd

4.0 nd 4.6 nd 0 0 0 0 0 0 0 0 0 0 0 1 0 0 nd 0 0 0 0.1 0 0 nd 0 0 nd 0 0 0 0 0.1 0 0 0 0 0 0 0 0 1 0.5 0 0 10 5 0 nd nd

nd nd 0 0 0 0 0 0 0 0 0 0 nd 0 0 0 0 0 0 0 0 0 0 nd nd nd nd nd 0 0 0 0 0 0 0 0 4 4 4

0 0 4 0 nd nd

0 nd 0 nd 0 0 0 0 0 0 0 0 0 0 nd 0 0 0 0 0 0 0 0 0 0 nd nd nd nd nd 0 0 0 0 0 0 0 0 0 0 20 20 45 32.5 2.7 0 0 60 30 0 nd nd

nd nd 0 0 0 0 0 0 0 0 0 0 nd 0 0 0 0 0 0 0 0 0 nd nd nd nd nd 0 0 0 0 0 0 0 0 4 nd 4

0 0 4 nd nd nd

1.7 nd 1.7 nd 0 0 0 0 0 0 0 0 0 0 nd 0 0 0 0 0 0 0 0 0 nd nd nd nd nd 0 0 0 0 0 0 0 0 0 0 40 nd 60 60 3.1 0 0 10 5 nd nd nd

4 ;(2) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 0 3 4 ; 4 4 4 4 2 4 4 4 0 nd 4

4 0 4 0 3 4

19.4 5 18.7 10 30 45 35 30 30 20 50 50 30 30 30 30 45 15 30 20 40 30 25 25 5 0 10 5 0 30 50 30 26.7 40 20 30 20 5 18.7 0 nd 35 35 25.8 20 0 30 15 0 15 15

D. Rubiales et al. / Crop Protection 46 (2013) 38e43

41

Table 2 (continued ) Accession

Donor’s code

Species

Subgenus/section

U. viciae-fabae ex V. faba

U. viciae-fabae ex L. culinaris

U. pisi

ITa

DSa

IT

DS

IT

DS

IT

DS

253 254 284

VIC 771/82 VIC 309/89 34 e VHy10

Vicia/Hypechusa Vicia/Hypechusa Vicia/Hypechusa

0 nd 0 0 0 0 0

97 98

VIC 475/89 VIC 474/78

Vicia/Hypechusa Vicia/Hypechusa

0 0

10 nd 0 5.0 nd 0 0 0 0 nd nd

0 0 0

Vicia/Hypechusa Vicia/Hypechusa Vicia/Hypechusa Vicia/Hypechusa

0 nd 0 0 nd 0 5 0 1,7 nd nd

4 nd 0

PI 201994 PI 249880 VIC 311/79 VIC 312/77

34 36

PI 220887 PI 220888

Vicia/Hypechusa Vicia/Hypechusa

0 0

Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae

0 0 0

33 37 44 45 104 105 106 107

PI 227472 PI 393824 PI 420425 PI 420424 VIC 315/79 VIC 657/79 VIC 765/79 VIC 796/79

Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae Vicia/Peregrinae

0 nd 0 0 0 0 0 0

233 237 77

VIC 66/86 VIC 71/88 PI 206780

Vicilla/Pedunculatae Vicilla/Cracca

0 0 nd

73 75 76 150 151 152 153 155 156

PI 298002 PI 298016 PI 449330 VIC 735/77 VIC 359/90 VIC 868/90 VIC 839/80 VIC 301/89 VIC 302/89

Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca

0 0 0 0 0 2 0 1 3

61 206 261

867891 VIC 654/79 VIC 782/80

Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca

4 4 0

32 35 272

PI 388742 PI388825 VIC 785/88

Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca

0 1þ 4

22 16 17 132 133 134 4 5

PI 308120 PI 206492 PI206493 VIC 843/79 VIC 637/74 VIC 633/74 PI 210431 PI 212044

Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca Vicilla/Cracca

0 0 0 0 0 0 0 nd

202 79 99 102

VIC 43/86 PI 212333 ERV 75/89 ERV 3/89

Cracca/Cassubicae Vicilla/Ervilia Vicilla/Ervilia Vicilla/Ervilia

0 2 nd 0

69 71 147 149

PI 244922 PI 449361 VIC 299/79 VIC 887/80

Vicilla/Ervoides Vicilla/Ervoides Vicilla/Ervoides Vicilla/Ervoides

0 0 ; 0

10 60 35.0 8.2 5 10 25 13.3 40 30 0 25 20 30 60 35 30 25.4 0 0 nd 0 nd 0 35 nd 50 40 20 10 50 30.7 25 nd 50 37.5 nd 10 40 25.0 nd 10 0 50 30 10 40 nd 23.3 26.7 nd 20 30 20 23.3 25 60 90 20

0 0

VIC 831/80 VIC 746/79 VIC 730/79

0 0 0 5.9 15 40 20 25 0 0 nd 0 0 nd 0 0 0 8.3 0 0 nd 0 0 0 60 nd 5 nd 0 10 nd 12.5 60 nd 35 47.5 nd 30 25 27.5 nd 0 0 0 0 0 0 nd 0 13.2 nd 40 nd 40 40 10 20 30 nd

4 4

220 257 267

0 nd 0 0 0 0 0 0 0 0 0 0 0 0 0 0.7 0 0 0

0 nd 0

11 12 108 109

V. hybrida L. V. hybrida L. V. hybrida L. Average V. lutea L. V. lutea L. V. lutea L. V. lutea L. Average V. melanops Sibth. & Sm. var. loiseani V. melanops Sibth. & Sm. var. melanops Average V. pannonica Crantz V. pannonica Crantz Average Average Section Hypechusa V. michauxii Spreng. V. michauxii Spreng. V. michauxii Spreng. Average V. peregrina L. V. peregrina L. V. peregrina L. V. peregrina L. V. peregrina L. V. peregrina L. V. peregrina L. V. peregrina L. Average Average Section Peregrinae V. onobrychioides L. V. cracca L. var. cracca V. cracca L. ssp. stenophyll Average V. benghalensis L. V. benghalensis L. V. benghalensis L. V. benghalensis L. V. benghalensis L. V. benghalensis L. V. benghalensis L. V. benghalensis L. V. benghalensis L. Average V. monantha Retz. V. monantha Retz. V. monantha Retz. Average V. palaestina Boiss. V. palaestina Boiss. V. palaestina Boiss. Average V. sicula (Raf.) Guss. V. villosa Roth V. villosa Roth V. villosa ssp. eriocarpa (Hausskn.) P.W. Ball V. villosa ssp. eriocarpa (Hausskn.) P.W. Ball V. villosa ssp. eriocarpa (Hausskn.) P.W. Ball V. villosa ssp. varia (Host) Corb. V. villosa ssp. varia(Host) Corb. Average Average Section Cracca V. cassubica L. V. ervilia (L.) Willd. V. ervilia (L.) Willd. V. ervilia (L.) Willd. Average Vicia articulata Hornem. Vicia articulata Hornem. Vicia articulata Hornem. Vicia articulata Hornem.

0 0 0 5.0 0 0 0 0 0 15 0 7.5 0 0 0 5.2 0 0 25 8.3 0 0 0 0 30 15 5 0 6.2 6.8 35 25 0 12.5 0 0 0 0 0 nd 0 0 nd 0 25 20 1 15.3 5 0 0 1.7 25 0 0 0 0 0 25 30 7.9 5.7 0 0 0 0 0 0 0 15 0

0 nd 0 0 0 0 0 0 0 0 0 0 nd 0 0 0 0 0 0 5 0 1 1 0.2 15 5 0 6.7 0 1 10 3.7 0 0 0 0 0 0 0 nd 0 1.6 0 5 nd 0 2.5 0 0 1 0

nd 0 2þ 0 nd nd 0 0

4 4 4 0 0 nd 0 0 nd 0 0

0 0 nd 0 0 4 nd 4 nd 0 4 nd 4 nd 4 nd 4 4 nd ; 0 ; 0 0 0 nd

nd 4 nd 4 4 4 4 nd

nd ; 0 0 nd nd

4 4 4 4 4 0 4 4 4 4 4

0 0 nd nd ; 4 nd 4 4 3 34 4 nd 4 nd 1þ 4 nd 1 0 4 4 2 4 nd

nd 4 4 4 4 4 4 4

U. viciae-fabae ex V. sativa

0 0 0 0 2þ 0

0 0 4 0 0 0 0 4 4 4 0

4 4 0 0 0 0 0 0 nd 0 0 nd 4 4 4 1 0 0 4 0 0 0 0 0 4 4

0 0 0 0 0 0 3 0

(continued on next page)

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D. Rubiales et al. / Crop Protection 46 (2013) 38e43

Table 2 (continued ) Accession

198 262 235 276 52 55 56 214 215 216 217 a

Donor’s code

Species

VIC 36/79 VIC 78/87 VIC 68/83 VIC 643/80 PI 183100 PI 420171 PI 422499 VIC 784/74 VIC 727/79 VIC 308/75 VIC 307/76

Average V. pisiformis L. V. unijuga A. Braun V. megalotropis Ledeb. V. loiseleurii (M. Bieb.) Litv. (sin V. meyeri) V. hirsuta (L.) Gray V. hirsuta (L.) Gray V. hirsuta (L.) Gray V. hirsuta (L.) Gray V. hirsuta (L.) Gray V. hirsuta (L.) Gray V. hirsuta (L.) Gray Average

Subgenus/section

Vicilla/Vicilla Vicilla/Vicilla Vicilla/Variegatae Vicilla/Ervum Vicilla/Ervum Vicilla/Ervum Vicilla/Ervum Vicilla/Ervum Vicilla/Ervum Vicilla/Ervum Vicilla/Ervum

U. viciae-fabae ex V. faba

U. viciae-fabae ex L. culinaris

U. pisi

U. viciae-fabae ex V. sativa

ITa

DSa

IT

DS

IT

DS

nd nd 0 0 0 0 0 nd 0 0 0

0.2 nd nd 0 0 0 0 0 nd 0 0 0 0

0 nd nd 0 ; nd ; 0 nd 1 ;

20 0 nd nd 0 0 nd 0 0 nd 5 0 0.8

0 nd nd nd 4 nd 4 4 nd 4 1

50 0 nd nd nd 10 nd 25 20 nd 20 5 16

IT

DS

nd 0 0 0 0 0 0 0 0 ; 0

4 nd 0 0 0 0 0 0 0 0 2 0 0.2

IT, infection type; DS, disease severity.

precise generic boundaries between these genera have been much debated which has led to an abundant and complex synonymy. The genus Vicia include many species of agricultural interest, such as faba bean and the various vetches that may be infected by rust (Sillero et al., 2006). The present study contributes to the clarification of host range of U. viciae-fabae and U. pisi, and confirms host specialization within U. viciae-fabae (Emeran et al., 2005; Barilli et al., 2009, 2012). It also identified sources of resistance to the various rusts that can be used in rust resistance breeding. The differential response of V. faba, V. sativa and L. culinaris checks confirms the existence of specialized isolates of U. viciaefabae as suggested by Emeran et al. (2005). Pea was more infected by U. pisi, but could also be infected by U. viciae-fabae isolates as already reported (Gäumann, 1959; Cummins, 1987; Barilli et al., 2012). Host range of these host specialized forms of U. viciaefabae was not so clear cut when we test other species of Vicia, however a tendency was observed that U. viciae-fabae ex V. faba infected only V. faba; that U. viciae-fabae ex V. sativa infected most species of sections Vicia and Narbonensis of subgenus Vicia, although also species of section Hypechusa and Peregrinae as well as others in subgenus Vicilla, mainly of sections Pedunculatae and Cracca. U. viciae-fabae ex L. culinaris infected species particularly of the sections Cracca, Ervilia and Ervoides of subgenus Vicilla, but also some of section Peregrinae of subgenus Vicia and to minor extent sections Vicia and Hypechusa. Similarly, U. pisi was less specialized; infecting species of the two subgenera, but in common with U. viciae-fabae ex L. culinaris, U. pisi infected mainly species of sections Cracca, Ervilia, Ervoides, and Ervum of subgenus Vicilla, as well as Peregrinae and Hypechusa of subgenus Vicia. The relevant difference was that U. pisi was the only of the studied rusts able to infect V. bithynica, V. pannonica and V. hirsuta. Knowledge of the host range is important to determine whether other crops could be affected. Furthermore, it is possible that alternative hosts could provide a means of overwintering of the pathogen, providing inoculums to initiate epidemics in future years. Conversely, different plant species could be infected by more than one rust species. In particular some accessions of V. hyrcanica, V. pyrenaica, V. michauxii, V. monantha, V. palaestina or V. articulata could be severely infected by up to three of the rusts studied. Similarly, grasses have been reported to be receptive to a wide range of rusts and this was considered common in arid regions where rust development is weak (Anikster and Wahl, 1979). The lack of strong preferential selection pressure would be responsible for the absence of differential reactions in the host to the various rusts. Such hosts might be suitable sites for somatic hybridization between rust, as already reported for varieties of Puccinia coronata (Anikster and Wahl, 1979).

The expansion of the host range of rust fungi is also important to the understanding of their phylogenetic history and sometimes sheds light on the evolution of the hosts. According to Dietel’s (1904) theory of host-rust coevolution, the transition to new hosts proceeds in the direction of the progressive evolution of the vascular plant or to plants that originated at the same time as the source host, but not to plants that had evolved in earlier periods. Evolution of varieties or formae speciales has frequently been acknowledged in rust complexes such as stem rust, leaf rust and crown rust complexes infecting cereals and grasses (Puccina graminis, Puccina triticina and P. coronata senso lato, respectively) as summarized by Anikster and Wahl (1979). Subgenus Vicia is smaller, less variable morphologically and comprises almost exclusively annuals including the most agriculturally important species as V. faba, V. sativa and V. narbonensis. V. faba is quite distinct and isolated from other Vicia taxa. It was earlier even considered a separate genus (Tournefort, 1694). Based on the taxa currently known, there remains no close ally of V. faba. It is relevant that U. viciae-fabae ex. V. faba was very specialized on faba bean only and that U. viciae-fabae ex V. sativa was specialized mainly on sections Narbonensis and Vicia of the most evolved subgenus Vicia. In contrast, species of subgenus Vicilla that is considered more primitive and diverse (Maxted, 1995) could be infected by less specialized rusts such as U. pisi and U. viciae-fabae ex L. culinaris. Similar trend was observed in species of section Hypechusa that is considered the most variable and less evolved group of the subgenus Vicia (Kupicha, 1976; Caputo et al., 2006) that was infected by most rusts. This trend suggests a possible cospeciation of rusts with host. Such co-speciation of rusts with the diversification of the host lineage has widely been acknowledged (Leppik, 1972; Anikster and Wahl, 1979; Savile, 1979) and has already been suggested with Uromyces rust in Fabaceae (Van der Merwe et al., 2008). Association of telial hosts of species of Uromyces on Fabaceae has pointed that Uromyces is a monophyletic group, which radiated on Fabaceae (Van der Merwe et al., 2007). However, there are many examples of jumps, suggesting that host transfer has probably been at least as frequent as co-speciation (Hart, 1988; Van der Merwe et al., 2008). Uromyces appendiculatus showed relationship with host phylogeny, whereas Uromyces vignae did not (Eckenwalder and Heath, 2001). Our results confirm the wide host range of U. pisi that besides pea can infect lentil and a number of Vicia species particularly of the Vicilla subgenus. The facts that U. pisi is the less specialized species and the one with the broadest host range (Barilli et al., 2012) suggest that other Uromyces species might have evolved from U. pisi. U. viciae-fabae ex V. faba is very specialized on V. faba only. There is a good reservoir of resistance in Vicia spp. both of hypersensitive

D. Rubiales et al. / Crop Protection 46 (2013) 38e43

and non-hypersensitive type available for faba bean resistance breeding once the barriers for crossability are surpassed. Conversely, U. viciae-fabae ex V. sativa showed a broad host range, infecting many species of subgenus Vicia not infected by any of the previous rusts (V. johannis, V. galilea, V. sativa, V. sepium, V. cordata, V. narbonensis or V. striata). In spite of the common susceptibility against this rust in the subgenus Vicia, interesting levels of resistance were identified in most species. Both hypersensitive resistance (low IT) and resistance based on a reduced DS in spite of a compatible interaction was identified. Acknowledgments This work was supported by the Spanish Ministry of Science and Innovation [AGL2008-01239 and 2011-22524, co-financed by FEDER], and Spanish Agency for International Cooperation (AECI project D/030364/10). References Anikster, Y., Wahl, I., 1979. Coevolution of the rust fungi on Gramineae and Liliaceae and their hosts. Annu. Rev. Phytopathol. 17, 367e403. Barilli, E., Moral, A., Sillero, J.C., Rubiales, D., 2012. Clarification on rust species potentially infecting pea (Pisum sativum L.) crop and host range of Uromyces pisi (Pers.) Wint. Crop Prot. 37, 65e70. Barilli, E., Satovic, Z., Sillero, J.C., Rubiales, D., Torres, A.M., 2011. Phylogenetic analysis of Uromyces species infecting grain and forage legumes by sequence analysis of nuclear ribosomal internal transcribed spacer region. J. Phytopathol. 159, 137e145. Barilli, E., Sillero, J.C., Serrano, A., Rubiales, D., 2009. Differential response of pea (Pisum sativum) to rusts incited by Uromyces viciae-fabae and U. pisi. Crop Prot. 28, 980e986. Caputo, P., Frediani, M., Venora, G., Ravalli, M., Ambrosio, M., Cremonini, R., 2006. Nuclear DNA contents, rDNAs, and karyotype evolution in subgenus Vicia: III. The heterogeneous section Hypechusa. Protoplasma 228, 167e177. Cummins, G.B., 1987. Rust Fungi on Legumes and Composites in North America. The University of Arizona Press, USA. Dietel, P., 1904. Betrachtungen über die verteilung der Uredineen auf ihren Nährpflanzen. Centralbl. Bakteriol. Parasitenkd. Infektionskr. Abt. 2 Allg. landwirtsch. Technol. Bakteriol. Gärungsphysiol. Pflanzenpathol. 12, 218e234. Emeran, A.A., Román, B., Sillero, J.C., Satovic, Z., Rubiales, D., 2008. Genetic variation among and within Uromyces species infecting legumes. J. Phytopathol.156, 419e424.

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