Joanna Marcinkowska1) Lech Boros2), Anna Wawer2)
1)
De part ment of Plant Pa thol ogy of War saw Uni ver sity of Life Sci ences, War saw, 02-870 Nowoursynowska 159, Po land, 2) De part ment of Seed Sci ence and Tech nol ogy of Plant Breed ing and Acclimatisation In sti tute,
Radzików, 05-870 B³onie, Po land
RESPONSE OF PEA (PISUM SATIVUM L.) CULTIVARS AND LINES TO SEED INFECTION BY ASCOCHYTA BLIGHT FUNGI
ABSTRACT
Seeds col lected from 10 cultivars and lines of Pisum sativum of both 'afila' and normal fo liage type, in oc u lated in field by Mycosphaerella pinodes and Phoma pinodella, were eval u ated for in ci dence of fungi re spon -si ble for ascochyta blight. Also seed ger mi na tion and con tam i na tion by other fungi were con -sid ered. Sur face ster il ized seeds were plated on Coon agar me dium. A sam ple con tains 50 seeds. Data were taken af ter 8 days of in cu ba tion. Fre quency of spe cies oc cur rence de pended not only on char ac ters of cultivars and lines, weather con di tions dur ing 1998-2001, but on in oc u la tion treat ment. Clear re sponse be tween cultivars and lines to tested fac tors was noted. Seeds of nor mal leaved line 344/87/3 and cv. Ru bin were the most in hab ited by all fungi, and ger mi nated very poorly. Seeds of eight cultivars and lines, in clud ing cvs: Kwestor, Agra, Miko, were less in fected and better ger mi nated. Al most 40% of nonger mi nated seeds were in hab ited by var i -ous fungi, but cultivars and lines dif fer ences were not clear. Seed in fec tion by ascochyta blight fungi was rather low, reach ing from 2.26 – 11.34 per cent, a mean 4.81, but al ways the high est for M. pinodes. Per cent age of con tam i nated seeds by other fungi, mainly saprobic, was al ways higher, even 5 times, with an av er age 16.73%. The best for seed ger mi na tion was grow ing sea son of 2000, while for seed in fes ta tion, nonger mi -nated and ger mi -nated, 2001.
Key words: ascochyta blight, oc cur rence, Pisum, seeds re ac tion, in ten sity
INTRODUCTION
Ascochyta blight caused by Mycosphaerella pinodes (Berk. et Blox.) Vesterg. (syn. Didymella pinodes (Berk. et Blox.) Petrak) anamorph
Ascochyta pinodes L. K. Jones, Ascochyta pisi Libert and Phoma pinodella
(L.K.Jones) Mor gan-Jones et Burch, is a se ri ous dis ease of Pisum sativum L., es pe cially in wet sea sons, of many coun tries over the world, like Can ada (Wallen 1974, Xue et al. 1997), Aus tra lia (Bretag et al. 1995), France (Roger et al. 1999), Po land (Marcinkowska 2002). The plants may be in -fected dur ing all de vel op ment stages (Law yer 1984, Marcinkowska 1996 b, Xue and Warkentin 2001). When seeds were in fected re duc tion in pea emer gence, and in con se quence grain yield losses oc curred (Bretag et al. Communicated by Ed ward Arseniuk
P L A N T B R E E D I N G A N D S E E D S C I E N C E
Volume 59 2009
1995, Xue et al. 1997). Dif fer ent tri als were per formed for find ing sources of re sis tance to the fungi re spon si ble for Ascochyta blight of pea (Ali et al. 1978, Clulow et al. 1991, Knappe and Hoppe 1995, Boros and Wawer 2007), but no body found sources of high re sis tance to the patho gens. In such sit u a tion re sponse of newly re leased pea lines and cultivars to Ascochyta blight fungi have been tested in some coun tries (Kraft et al. 1998, Xue and Warkentin 2001, Zhang et al. 2006). Also in Po land re cently bred pea cultivars and lines were tested against fungi re spon si ble for
ascochyta blight. The stud ies were done on plants in fields (Boros and
Marcinkowska, 2007 in print) and on seeds since level of their in fec tion can in di cate on ge no type sus cep ti bil ity (Marcinkowska 1996a). The in ves ti ga -tion of occurrence fre quency of fun gal spe cies caus ing Ascochyta blight, es pe cially A. pinodes and P. pinodella, on seeds of new Pol ish cultivars and lines was the main goal of the pre sented work. Abil ity of seed ger mi na tion and their con tam i na tion when nonger mi nated and ger mi nated was also re -ported.
MATERIALS AND METHODS
In the veg e ta tion sea sons of 1998-2001 pea plants of four cultivars: Agra, Kwestor, Miko, Ru bin, and 6 lines: 1528/96, 344/87/3, 1166/96, RAH 796, RAH 997, RAH 897, both fo liage type, grow ing on plots in fields of Plant Breed ing and Ac cli ma ti za tion In sti tute (PBAI) at Radzików (Cen tral-East Po land) were in oc u lated prior to flow er ing stage with conidial sus pen sion (2 ×106 spores in 1ml of wa ter) of A. pinodes, P. pinodella and mix ture of both spe cies (Boros and Marcinkowska 2007). Plots of these three treat ments and wa ter, as con trol, were rep li cated four times. Af ter har vest a sam -ple of 50 seeds was col lected from each plot for eval u a tion. To tally in a year 800 seeds of each ge no type were tested from Jan u ary to March.
Seeds were sur face ster il ized as fol lows: 30 sec. wash ing in wa ter with de ter gent, 60 sec. rins ing un der tap wa ter, 15 sec. keep ing in 96% ethyl al -co hol, and 15 sec. in 15% H2O2, 5sec. rins ing in dis tilled ster il ized wa ter. Ster il ized seeds were placed into 4 Petri plates (Pp) of 10 cm di am e ter, 2 Pp with 12 and 2 with 13 seeds on Coon’s (CN) agar me dium (Ali et al.1978). Num ber of non-ger mi nated and ger mi nated seeds, healthy and in fected by
M. pinodes, A. pisi and P. pinodella, and those trans mit ting all other fungi
was counted af ter (8) eight days from the time the plates were ex posed for three 3 days un der dis persed nat u ral day light and next four (4) days to bulbs with flu o res cent day light for 14 four teen hours a day. Fun gal spe cies iden -ti fi ca -tion pro ce dure was as pre vi ously de scribed (Marcinkowska 1997).
Num ber non-ger mi nated and in fected by fungi seeds was ex pressed in per cent of 50 seeds in a sam ple. For sta tis ti cal eval u a tion percents were trans formed ac cord ing to Bliss. The GLM pro ce dure was used to an a lyze the dif fer ences among a fac tor (SAS 2004).
RESULTS
Dif fer ences in the abil ity of seed ger mi na tion and fun gal oc cur rence on them were found be tween tested cultivars and lines, treat ments and years of the per formed stud ies. When cultivars and lines were com pared over years and treat ments, the high est (23.4) mean per cent of non-ger mi nated seeds was noted for line 344/87/3 and the next (17.4) for cv. Ru bin of the same ho -mog e nous group (Table 1). The other eight cultivars and lines be longed to
the sec ond group, with the low est (8.0) per cent age for RAH 796 and next (9.2) for Miko. It was de pend ing on cultivars and lines but per cent age of healthy nonger mi nated seeds was al ways higher than for seeds trans mit -ting fungi. A mean of non-ger mi nated seedfor all cultivars and lines was 12.3%. Fre quency of fungi oc cur rence on ger mi nated seeds reached on an av er age 16.2 per cent and 4.9 for nonger mi nated Per cent age of nonger mi -nated seeds trans mit ting fungi was cov ered by 4 ho mog e nous groups from 2.2 for RAH 796 to 9.6 for Ru bin. The dif fer ences be tween cultivars and lines for per cent age of ger mi nated seeds on which oc curred fungi were sta -tis ti cally sig nif i cant (3 ho mog e nous groups). The high est (23.5) per cent age of ger mi nated seeds in hab ited by fungi was for line 344/87/3 and the next (21.8) of the same group was cv. Ru bin. The low est (12.5) per cent age was
Ta ble 1 Abil ity of ger mi na tion and fungi oc cur rence on seeds of tested cultivars and lines
(mean from treat ments and years)
Genotype
Non-germinated seeds Seeds transmitting fungi Non-germinated Germinated Percent* HG Percent* HG Percent* HG
1528/96 10.5 b 4.7 b, c, d 12.8 b, c Miko 9.2 b 3.8 d, c 15.1 b, c 344/87/3 23.4 a 6.0 a, b, c, 23.5 a 1166/96 10.3 b 7.3 a, b, 16.4 b Agra 11.7 b 4.3 b, c, d 16.1 b RAH 796 8.0 b 2.2 d 12.5 c RAH 997 11.3 b 3.6 a, b 16.6 b Rubin 17.4 a 9.6 a 21.8 a Kwestor 11.0 b 3.7 b, c, d 14.3 b, c RAH 897 10.2 b 4.0 b, c, d 13.1 b, c Mean 12.3 4.9 16.2
per cent of a sam ple;
for RAH 796 and in the same group were also lines 1528/96, RAH 897 and cvs Kwestor and Miko.
When treat ments were con sid ered, sta tis ti cally sig nif i cant dif fer ences ex isted for all non-ger mi nated seeds, and also those trans mit ting fungi (Table 2). The high est, re spec tively 16.2 and 6.9 per cent age was for seed sam ples de rived from con trol treat ment. Over dou ble more non-ger mi nated seeds were healthy com par ing with non-ger mi nated but trans mit ting fungi. Per cent age of ger mi nated seeds trans mit ting fungi did not dif fer sig nif i -cantly be tween treat ments and was from 2 to 4 times higher than for non-ger mi nated.
The dif fer ences in seed ger mi na tion and fungi oc cur rence on them were sta tis ti cally sig nif i cant be tween years (Table 3). Two ho mog e nous groups were dis tin guished for nonger mi nated seeds, with the high est (15.7) per cent in 1998 and the low est (9.8) in 2000. Each year oc curred less nonger
-Ta ble 2 In flu ence of field treat ments on de rived seed ger mi na tion and trans mis sion of fungi
in de pend ently on ge no types and years
Treatments
Non-germinated seeds
Seeds transmitting fungi Non-germinated Germinated
Percent* HG Percent* HG Percent* HG
Control 16.2 a 6.9 a 14.4 a
Ascochyta 9.5 b 4.2 b 15.6 a
Phoma 11.3 b 4.1 b 19.1 a
Asc.+Ph. 12.1 ab 4.7 b 15.9 a
ex pla na tion — see Table1; HG – ho mog e nous groups
Treat ments: Ascochyta – in oc u lated with A. pinodes; Phoma – in oc u lated with P. pinodella; Asc.+Ph. – in oc u -lated with mix ture of A. pinodes and P. pinodella
Ta ble 3 In flu ence of year on seed ger mi na tion and fungi oc cur rence in de pend ently on ge no types and treat ments
Year
Non-germinated seeds
Seeds transmitting fungi Non-germinated Germinated
Percent* HG Percent* HG Percent* HG
1998 15.7 a 5.9 b 12.3 b
1999 11.4 a, b 3.1 c 6.6 c
2000 9.8 b 1.8 c 13.1 b
2001 12.3 a, b 9.2 a 32.9 a
ex pla na tion — see Ta ble 1; HG – ho mog e nous groups
mi nated seeds trans mit ting fungi than healthy and ger mi nated with fungi, reach ing the high est (9.2) per cent age in 2001 and the low est (1.8) in 2000. Data con cern fun gal oc cur rence on ger mi nated seeds rep re sented also 3 ho -mog e nous groups, with the high est (32.9) per cent found as well in 2001 and the low est (6.6) in 1999.
On seeds of tested cultivars and lines oc curred 18 dif fer ent spe cies of fungi. Eight of them have been patho genic for plants, with M. pinodes, A.
pisi, P. pinodella and Fusarium solani (Mart.) Sacc., spe cific patho gens for
pea, while Bo try tis cinerea Pers.: Fr., Stemphylium botryosum Wallr.,
Sclerotinia sclerotiorum (Lib.) de Bary and Rhizoctonia solani Kühn have
been polyhagous spe cies. In the group of patho genic spe cies dom i nated fungi re spon si ble for ascochyta blight of pea, the other five spe cies in fected seeds only spo rad i cally. Ma jor ity of seeds in the group of other fungi were con tam i nated by saprobic fungi and only a few in fected by patho genic spe -cies, but never by re spon si ble for ascochyta blight, so the other fungi mainly con sisted of saprobes. Per cent age of saprobic fungi was much higher to ascochyta blight in fected seeds (Fig. 1) for each cultivar, and for a mean (16.73) independly on ge no type, was over three times more (4.81).
Among the saprobic spe cies Alternaria alternata (Fr.) Keissler oc curred on seeds as the most com mon and out of Fusarium spp. most of ten F. poae
(Peck) Woll. was iso lated. F. avenaceum (Corda: Fr.) Sacc.,
F. sporotrichoides and not iden ti fied Fusarium iso lates were ob tained, be
-sides Ulocladium chartarum, Chaetomium spp., Cladosporium spp.,
Penicillium spp., Aspergillus spp., Mucor spp. Out of ascochyta blight
fungi A. pinodes was iso lated most of ten, since a mean for cultivars and
Fig. 1. Occurance of Ascochyta blight fungi (A.pisi, M. pinodes and P. pinodella) and other spe cies on seeds of tested cultivars and lines (Mean over treat ments and a years)
lines, in de pend ently on treat ments and years, was 3.44 per cent of a sam ple, the next was P. pinodella with 1.16 per cent and A. pisi oc curred only spo -rad i cally as on 0.21 per cent of a seed sam ple (Ta ble 4).
There were not sig nif i cant dif fer ences be tween per cent age of in fected seeds by the last fun gus in a sam ple, nei ther for nonger mi nat ing nor ger mi -nat ing in fected seed. In fec tion of seeds by A. pinodes dif fered sta tis ti cally and for a whole sam ple 2 ho mog e nous groups were dis tin guished like for in fected ger mi nat ing seeds, but for non-ger mi nat ing 3 groups cov ered the
Ta ble 4 Oc cur rence of fungi on seeds of the tested cultivars and lines (mean from treat ments and years)
Treatments
Percent of sample
Homogenous groups for Percent of sample
Homogenous groups for Sample Non-germ. Germ.seeds Sample Non-germ. Germ.seeds
Ascochyta pisi Ascochyta pinodes
1528/96 0.22 a a a 1.67 b c b Miko 0.22 a a a 4.06 b a, b, c b 344/87/3 0.37 a a a 8.15 a a, b a 1166/96 0.24 a a a 3.18 b a, b, c b Agra 0.24 a a a 2.68 b c b RAH 796 0.19 a a a 1.81 b c b RAH 997 0.03 a a a 1.61 b b, c b Rubin 0.31 a a a 7.31 a a a Kwestor 0.06 a a a 1.89 b b, c b RAH 897 0.21 a a a 2.05 b b, c b Mean 0.21 3.44
Phoma pinodella Other fungi
1528/96 0.77 b a, b c 15.35 b, c, d a, b b, c Miko 0.15 b a, b c 13.52 d, c b b, c 344/87/3 2.82 a a, b a 18.97 a, b a, b a 1166/96 1.39 b a, b b, c 19.18 a, b a a, b Agra 1.11 b a, b c 17.57 a, b, c a, b a, b, c RAH 796 0.37 b b c 12.95 d a, b c RAH 997 0.62 b b c 17.95 a, b, c a, b a, b Rubin 3.12 a a a, b 21.38 a a, b a Kwestor 0.71 b a, b c 15.66 b, c, d a, b a, b, c RAH 897 0.55 b b c 14.72 b, c, d a, b b, c Mean 1.16 16.73
data. The most in fected (8.15 and 7.31 per cent) by this fun gus were line 344/87/3 and cv. Ru bin, rep re sent ing the same group for a sam ple and sep a -rately on non-ger mi nated and ger mi nated seeds. The low est (1.61, 1.67, 1.81, 1.89) per cent age of in fected seeds in a sam ple were found re spec -tively for lines: RAH 997, 1528/96, RAH 796 and cv. Kwestor. The high est (3.12 and 2.82) per cent of in fec tion in a seed sam ple of cv. Ru bin and line 344/87/3 was also noted for P. pinodella and both cultivars and lines rep re -sented the same ho mog e nous group. The sec ond group con tained eight other cultivars and lines with the low est per cent age 0.15, 0.37, 0.55, 0.62, re spec tively for Miko, RAH 796, RAH 897 and RAH 997. Per cent age of in fected nonger mi nated seeds by the last patho gen was also cov ered by 2 ho -mog e nous groups, but in case of ger mi nated in fected seeds 3 groups were dis tin guished. Apart from the num ber of groups, Ru bin and 344/87/3 were al ways the most in fected by P. pinodella. Dif fer ences in other spe cies oc -cur rence on seeds of tested cultivars and lines were not very clear but also sta tis ti cally doc u mented. The clear est data were for ger mi nat ing seeds for which cv. Ru bin and line 344/87/3 were con tam i nated by the high est per -cent age of dif fer ent fungi, but lines: RAH 796, 1528/96 and cv. Miko by the
low est.
When seed in fec tion of tested cultivars and lines was con sid ered to gether for three fun gal spe cies re spon si ble for ascochyta blight in de pend ently on treat -ments and years, line 344/87/3 and cv. Ru bin were the most in fected, reach ing 11.34 and 10.74 per cent of such seeds in a sam ple (Fig. 1). Also the per cent of oc cur rence of other fungi spe cies was the high est for both these cultivars and
Ta ble 5 Oc cur rence of fungi on seeds of dif fer ent treat ments (Means for cultivars/lines and years)
Treatments
Sample total Non-germinated Germin ted Sample total Non-germinated Germinated
Percent* HG HG HG Percent* HG HG HG
Ascochyta pisi Ascochyta pinodes
Control 0.26 a a a 2.70 b b b
Ascochyta 0.34 a a a 4.97 a a a
Phoma 0.12 a a a 2.78 b b b
Asc.+Ph. 0.17 a a a 3.26 a, b a, b a, b
Phoma pinodella Other species
Control 0.72 c b c 18.19 a a a
Ascochyta 0.48 c b c 14.66 a a a
Phoma 2.50 a a a 18.02 a a a
Asc.+Ph. 1.46 b a, b b 16.03 a a a
* Ex pla na tion — see ta ble 1; HG — ho mog e nous groups
Treat ments: Ascochyta – in oc u lated with A. pinodes; Phoma – in oc u lated with P. pinodella; Asc.+Ph. – in oc u -lated with mix ture of A. pinodes and P. pinodella
lines. Lines RAH 997, RAH 796, 1528/96 and cv. Kwestor showed the low est per cent age of in fec tion, from 2.26 to 2.66. Oc cur rence of other spe cies was the low est for line RAH 796 and cv. Miko.
Oc cur rence of fun gal spe cies dif fered be tween treat ments the seed de -rived from, but sig nif i cant dif fer ences were only noted for A. pinodes and
P. pinodella (Table 5). For the first spe cies 2 ho mog e nous groups were dis
tin guished for a whole seed sam ple, in fected nonger mi nated as well as ger -mi nated ones. The high est per cent of in fected seeds was found for Ascochyta treat ment. The data ob tained for mix ture of Ascochyta and
Phoma were lower, but cov ered by both, Ascochyta and Phoma treat ment
groups. The low est val ues showed con trol treat ment, but a lit tle higher seed in fec tion by A. pinodes was for Phoma treat ment. Dif fer ences in per cent of seed in fected by P. pinodella were cov ered by 2 groups for nonger mi nat -ing seeds and 3 for a whole sam ple and ger mi nated seeds. Treat ment with Phoma dis tin guished it self from oth ers by the high est seed in fec tion with P.
pinodella for a whole sam ple and sep a rately nonger mi nat ing and ger mi
nat ing seeds. The next less in fected seeds with P. pinodella were for treat -ment of Ascochyta and Phoma mix ture. The low est per cent age was found for Ascochyta, but data of this treat ment did not sig nif i cantly dif fer from
those of con trol one.
Per cent age of pea seed in fected by spe cies re spon si ble for Ascochyta blight and also oc cur rence of other spe cies of fungi on tested seeds dif fered sta tis ti cally for grow ing sea sons of 1998-2001 (Table 6). The high est (0.53) per cent of in fected seeds with A. pisi was ob served in 1998, but for
Ta ble 6 Oc cur rence of fungi on seeds dur ing 1998 – 2001 in de pend ently on cultivars and lines and treat ments
Years
Sample total Non-germinated Germinated Sample total Non-germinated Germinated
Percent* HG HG HG Percent* HG HG HG
Ascochyta pisi Ascochyta pinodes
1998 0.53 a a a 2.48 b a, b b
1999 0.11 b b, c b 1.13 c c c
2000 0.03 b c b 1.99 c b, c c
2001 0.21 b b a, b 6.62 a a a
Phoma pinodella Other species
1998 0.93 b b b 12.89 b b b
1999 0.61 b b b 7.89 c b, c c
2000 0.73 b b b 12.44 b c b
2001 2.88 a a a 33.7 a a a
A. pinodes and P. pinodella in 2001, reach ing re spec tively 6.62 and 2.88. In
the same sea son oc cur rence of other fungi on seeds was also the high est (33.7). All fun gal spe cies oc curred less fre quently in 1999 and 2000, when tem per a tures, es pe cially in June, were higher to an av er age of the pe riod from 1955 – 1996 (Fig. 2). In June 1999 to tal rain fall was over dou ble to those of 1955 – 1999, on the con trary to 2000, which was par tic u larly dry, only with 14.4 cm3 of rain falls. Spring of 2000 was the dri est and the hot test of the four grow ing sea sons, reach ing higher air tem per a tures and lower,
even ex tremely low, rain falls in April and June. DISCUSSION
Oc cur rence fre quency of ascochyta blight fungi on seeds ob tained at this study was in flu enced not only by ge no type and weather of the grow ing sea -son (Marcinkowska 1996a, b; Fougereux et al.2006), but also a way of pea plant in oc u la tion (treat ment) done on plots in field (Boros and Marcinkowska, in press). On seeds col lected from plants of plots in oc u lated with A. pinodes dom i nated just this spe cies, and on in oc u lated with P.
pinodella also the same last spe cies. Sec ond place in seed in fec tion in ten
-sity took re sults of in oc u la tion with mix ture of A. pinodes and P. pinodella, and the third place of A. pinodes. These re sults looked log i cally since A.
pinodes or P. pinodella in tro duced on plants dur ing in oc u la tion gave a par
-tic u lar spe cies pri or ity for de vel op ment in the veg e ta tion sea son, on all parts of pea so also on pods and seeds. How ever, re sults of in oc u la tion done with mix ture of both spe cies in di cated on in ter ac tion of A. pinodes and P.
pinodella, the phe nom e non re cently stud ied by Le May et al. (2006). On the
other hand, the ob tained re sults could also be partly in flu enced by spores of these spe cies liv ing in na ture. Like it hap pened for A. pisi. Even plants were not in oc u lated by A. pisi seed in fec tion by this spe cies was noted, for sure, by its conidia ex ist ing in an air. This fun gus oc curred on seeds, but only spo rad i cally, since it was not in tro duced by in oc u la tion and be cause grow -ing sea sons, es pe cially of 1999 and 2000, were in gen eral dry and hot, so were harm ful for plant in fec tion by dif fer ent patho gens.
Harm ful ef fect of weather on fun gal dis ease de vel op ment was also proved by re sults of seed in fec tion by A. pinodes and P. pinodella, the spe -cies in tro duced on plants. The in oc u lated plants af ter in cu ba tion pe riod were mainly ex posed to con di tions of ex ist ing weather and as the con se -quence per cent age of seeds in fected by spe cies used for in oc u la tion were quite low. The re sults de pended on years be ing the low est for 1999 and 2000, the years of too high tem per a tures and low rain falls by the end of veg -e ta tion p-e riod, th-e tim-e wh-er-e pods and so s-e-eds hav-e b-e-en in th-e stag-e of rip en ing (Fig. 2). Ac cord ing to Roger et al. (1999) tem per a ture over 20°C de creased de vel op ment of M. pinodes, while the dis ease was able to de -velop within a few days un der fa vor able mois ture and over a wide range of tem per a tures (15-25°C). On the other hand such weather in creased abil ity of seeds for ger mi na tion. Even the oc cur rence fre quency of A. pinodes and
P. pinodella on seeds were not high, the re sults in di cated on dif fer ent re
-sponse of ten stud ied cultivars and lines to A. pinodes and P. pinodella, the two spe cies mainly re spon si ble for pea ascochyta blight. The most in fected seeds by all three spe cies were for cv. Ru bin and line 344/87/3 and also their ger mi na tion was the poor est. Seeds of these cultivars and lines were also in -hab ited by the high est per cent of all other fungi.
Plants of line 344/87/3 had long veg e ta tion pe riod, high growth so could eas ily lodged and those char ac ters caused they were much stron ger in -fected. Also Wang et al. (2006) stated that lodg ing in creased se ver ity of
ascochyta blight caused by M. pinodes on field pea. The eight other
cultivars and lines be longed to the same group of low seed in fec tion and their abil ity for ger mi na tion was much better to noted for line 344/87/3 and Ru bin. Among them were lines: RAH 997, 796, cvs: Kwestor and Miko. For the last cultivar Xue and Warkentin (2001) found par tial re sis tance to stem and pod in fec tion by M. pinodes. Low seed in fec tion by ascochyta blight fungi noted in this study for Miko and al ready stated re sis tance of this cultivar to M. pinodes (Xue and Warkentin 2001) might be a prove that seed re acted to patho gens sim i larly like other parts of plant. Li Yonghao et al. (1992) found highly sig nif i cant re la tion ship be tween re sis tance of bean pods and leaves to Colletotrichum lindemuthianum (Sacc. Et Magn.) Bri. Et Cav. while Marcinkowska and Borucka (2001) re ported sig nif i cant re la tion ship be tween bean in fec tion by C. lindemuthianum in fields and the fun -gus in ci dence in seeds. Pos i tive cor re la tion be tween seed in fec tion by
ascochyta blight fungi and in ten sity of the dis ease on leaves, stems and
pods dur ing veg e ta tion re ported Marcinkowska (1996a) only when in ten -sity of blight in field was at least slight to mod er ate. But Bretag et al. (1995b) stated no cor re la tion be tween the level of seed in fes ta tion by M.
pinodes and the se ver ity of the dis ease. These au thors re ported only sig nif i
-cant re duc tion in pea emer gence when the level of seed in fec tion was over 11%. How ever, Bretag et al. (1995a) noted close cor re la tion be tween dis -ease se ver ity and re duc tion in grain yield.
In the per formed stud ies even plants were in oc u lated with both fungi, seed in fec tion of all cultivars and lines by M. pinodes oc curred more of ten than by P. pinodella. Those re sults have been in agree ment with ear lier (Marcinkowska 1996 b) re ported prev a lence of M. pinodes on seeds, leaves, stems and pods but the low est seed in ci dence of the sec ond spe cies. More over, oc cur rence fre quency of both spe cies in the pre sented stud ies, though they were in tro duced on plants, was lower com pared to per cent age of in fected seeds in years of early ninties (Marcinkowska 1996a,b). The dif -fer ences might be caused not only by dif -fer ent re ac tion of an other cultivars tested but also weather con di tions, very im por tant for fun gal dis eases de -vel op ment.
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