Lentil (Lens culinaris Medic.) is one of the oldest known protein-rich food legumes. Lentil is the second pulse crop after dry bean in Bulgaria. Diseases such as Ascochyta blight and Lentil wilt play a major role in reducing lentil yield. Thirty two lentil genotypes with different geographical origin were screened for reaction to Fusarium oxysporum f.sp. lentis during 2003-2004 from the Institute for Plant Genetic Resourses, Sadovo under greenhouse conditions. Three of the studied accessions (91-001, 91-028 and 98-001) were susceptible with 45 and 50 % of total wilted plant.Лещата (Lens culinaris Medic.) е една от най-старите зърнено бобови култури богати на протеин. Болести като чернилка /Ascochyta blight/ и фузарийно увяхване /Fusarium oxysporum f.sp. lentis/ играят важна роля в редуциране добива при тази култура. Тридесет и два образци с различен географски произход бяха изпитани срещу причинителя на фузарийното
увяхване. Изследването се проведе през 2003-2004 при условията на изкуствен инфекциозен фон. Три от образците (91-001, 91-028 и 98-001) представляват интерес за включване в селекционния процес

Peter Chavdarov

Tzvetelina Stoilova

English

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ORIGINAL PAPER121Volume 7 (2006) No. 1 (121-126)EVALUATION OF LENTIL GERMPLASM FOR DISEASE RESISTANCE TO FUSARIUM WILT (FUSARIUM OXYSPORUM F.SP. LENTIS)УСТОЙЧИВОСТ НА ОБРАЗЦИ ЛЕЩА КЪМ ФУЗАРИЙНО УВЯХВАНЕ (FUSARIUM OXYSPORUM F.SP. LENTIS)Tzvetelina Stoilova and Peter ChavdarovInstitute for Plant Genetic Resources, 4122 Sadovo, Bulgariae-mails: tzvets@hotmail.com, chavdarovpetar@abv.bgManuscript received: February 15, 2006; Reviewed: February 23, 2006; Accepted for publication: March 13, 2006ABSTRACTLentil (Lens culinaris Medic.) is one of the oldest known protein-rich food legumes. Lentil is the second pulse crop after dry bean in Bulgaria. Diseases such as Ascochyta blight and Lentil wilt play a major role in reducing lentil yield.Thirty two lentil genotypes with different geographical origin were screened for reaction to Fusarium oxysporum f.sp. lentis during  2003-2004 from the Institute for Plant Genetic Resourses, Sadovo under greenhouse conditions.Three of the studied accessions (91-001, 91-028 and 98-001) were susceptible with 45 and 50 % of total wilted plant.KEYWORDS: Lentil, evaluation, disease, Fusarium, germplasmРЕЗЮМЕЛещата (Lens culinaris Medic.) е една от най-старите зърнено бобови култури богати на протеин. Болести като чернилка /Ascochyta blight/ и фузарийно увяхване /Fusarium oxysporum f.sp. lentis/ играят важна роля в редуциране добива при тази култура.Тридесет и два образци с различен географски произход бяха изпитани  срещу причинителя на фузарийното увяхване. Изследването се проведе през 2003-2004 при условията на изкуствен инфекциозен фон. Три от образците (91-001, 91-028 и 98-001)  представляват интерес за включване в селекционния процес. КЛЮЧОВИ ДУМИ: леща, оценка, болести, фузариум, генплазма122 Journal of Central European Agriculture Vol 7 (2006) No 1Tzvetelina Stoilova and Peter ChavdarovINTRODUCTIONLentil (Lens culinaris Medic.) is one of the oldest known protein-rich food legumes. Lentil is also called “poor man’s meat” [2]. Lentil seeds are reach in protein, their mean value is at about 28,5% [14]. Lentil is the second pulse crop after dry bean in Bulgaria. It is grown mainly in North-Eastern part of the country. Owing to biotic and abiotic stresses, the crop yields is below attainable levels. Among the biotic factors diseases are serious threat to lentil production in many parts of the world. Lentil (Lens culinaris Medic.) suffers from a number of diseases which are caused by fungi, bacteria, viruses, nematodes and plant parasites [9]. Diseases such as Ascochyta blight and Lentil wilt play a major role in reducing lentil yield [6]. Lentil wilt, caused by Fusarium oxysporum f.sp lentis is one of the main limiting factors to successful cultivation. The effects of various environmental factors on pathogen growth and disease expression have been studied [13], [4]. At ICARDA, sources of wilt resistance have been identifi ed. These were distributed in the Lentil International Fusarium Wilt Nursery (LIFWN) for testing under fi eld conditions at different locations.  Mihov et al., (1987) [12], reported four cultivars with resistance under fi eld conditions in Bulgaria.   This pathogen is responsible for severe grain losses [5], [10], [1].  The disease appears either in the early stage of crop growth (seedling wilt) or during reproductive growth (adult plant wilt) [11]. Environmental conditions have always been a major constraint for stable lentils yields [16]. Moderately high soil temperature  (20 to 25° C) which favor fungal growth and sunlight, which enhances transpiration, seem to be the key factors determining symptom expression. The objective of this study was to identify the availability of resistance among 32 lentil genotypes to Fusarium oxysporum f.sp lentis under glasshouse conditions.MATERIAL AND METHODSThirty two lentil genotypes with different geographical origin (were screened for reaction to Fusarium oxysporum f.sp. lentis during  2003-2004 from the Institute for Plant Genetic Resourses, Sadovo under greenhouse conditions.Pure culture of the lentil pathogen are maintained in autoclaved soil [15], and then mixed with the planting soil [8]. Seeds from each accession were sown in a dish with size 30-30-10cm. The spore concentration of 2.5 x 102macroconidia ml-1 was used to inoculate sterilized fi eld soil in the greenhouse to screen for wilt. The resistance to Fusarium oxysporum f.sp. lentis was established by method of Dobrev, 1987, [3]. Disease symptoms on each plant were assessed two times per week till fl owering stage. The observations on seedling/plant mortality are converted to percent wilt for use on a 1 to 5 where: 1(I) = 0% infection, 2 (R) = 1 to 15% plants wilted, 3 (MS) = 15 to 25%plants wilted, 4 (S) = 25 to 50%, 5 (HS) =50 % or more plants wilted. In reporting resistance, it is very important to mention the crop growth stage of the screening. More lentil accessions, exhibiting resistance at the seedling stage, lose their resistance at the adult stage as it was reported from ICARDA (1990), [7].RESULTS AND DISCUSSIONSIn their resistance to Fusarium oxysporum f.sp. lentis under glasshouse conditions, 9 accessions, namely  91-009, 91-011, 93-002, 93-040, 95-005, 98-001, 98-013, 2000LEN464, 2000LEN466, were with seedling mortality to 15% during all the experiment, 2003-2004 (Table 1, 2). Accessions numbers: 93-002 (Russia) from 2004, 95-005 (Greece), 91-011(Germany) and 2000LEN466 (Syria) from 2003 were free from disease at seedling stage.  Highly susceptible (HS) with 50% or more at this Fig. 1.  Lentil plants, 1st free of symptoms and 2nd with symptoms of Fus. ox. f. sp. lentis EVALUATION OF LENTIL GERMPLASM FOR DISEASE RESISTANCE TO FUSARIUM WILT (FUSARIUM OXYSPORUM F.SP. LENTIS)123J. Cent. Eur. Agric. (2006) 7:1, 121-126Table. 1  Resistance to Fusarium oxysporum f. sp. Lentis of Lentil genotypes, 2003 �.� Cat.� SownseedsSeedlings Mortality seedl. ns.          % Mortality plants Ns                % Total  mortality  pl. Ns.                      % 1 2 3 4 5 6 7 8 9 101 302 20 10 10 50.0 2 20.0 12 60.0 2 345 20 3 17 85.0 2 66.7 19 95.0 3 82-001 20 3 17 85.0 1 33.3 18 90.0 4 82-003 20 6 14 70.0 4 66.7 18 90.0 5 91-001 20 17 3 15.0 8 47.0 11 55.0 6 91-002 20 16 4 20.0 9 56.2 13 65.0 7 91-006 20 0 20 100.0 - - 20 100.0 8 91-009 20 19 1 5.0 13 68.4 14 70.0 9 91-010 20 11 9 45.0 5 45.5 14 70.0 10 91-011 20 20 0 0.0 18 90.0 18 90.0 11 91-017 20 4 16 80.0 1 25.0 17 85.0 12 91-028 20 13 7 35.0 3 23.1 10 50.0 13 93-002 20 19 1 5.0 12 63.2 13 65.0 14 93-021 20 2 18 90.0 1 50.0 19 95.0 15 93-024 20 6 14 70.0 2 33.3 16 80.0 16 93-040 20 17 3 15.0 11 64.7 14 70.0 17 93-046 20 7 13 65.0 6 85.7 19 95.0 18 93-050 20 3 17 85.0 3 100.0 20 100.0 19 93-051 20 13 7 35.0 10 76.9 17 85.0 20 93-058 20 3 17 85.0 3 100.0 20 100.0 21 95-003 20 8 12 60.0 8 100.0 20 100.0 22 95-005 20 20 0 0.0 13 65.0 13 65.0 23 98-001 20 17 3 15.0 7 41.2 10 50.0 24 98-004 20 11 9 45.0 8 72.7 17 85.0 25 98-005 20 17 3 15.0 10 58.8 13 65.0 26 98-007 20 12 8 40.0 12 100.0 20 100.0 27 98-011 20 14 6 30.0 11 78.6 17 85.0 28 98-013 20 15 5 25.0 8 53.3 13 65.0 29 2000 LEN 463 20 11 9 45.0 6 54.5 15 75.0 30 2000 LEN 464 20 15 5 25.0 8 53.3 13 65.0 31 2000 LEN 465 20 0 20 100.0 - - 20 100.0 32 2000 LEN 466 20 20 0 0.0 12 60.0 12 60.0 33 2000 LEN 468 20 6 14 70.0 3 50.0 17 85.0 34  2000 LEN 470 20 13 7 35.0 5 38.5 12 60.0 35  2000 LEN 471 20 6 14 70.0 4 66.7 18 90.0 36 2000 LEN 472 20 11 9 45.0 5 45.5 14 70.0 stage were 16 of studied accessions with 60, 75, 80 and 90% mortality seedling plants. The genotypes 91-006 (Hungary) and 2000LEN465 (Syria) were classifi ed as highly susceptible with 100% of mortality seedling plants during both years.The accessions 93-050 and 93-058 showed highly susceptible during all the experiments in the next phase (adult plants) before fl owering, the remaining plants were mortality (Table. 1, 2). The accessions 345, 82-003, 91-002, 91-009, 91-011, 93-002, 93-040, 93-046, 93-051, 95-005, 98-004, 98-005, 98-011, 98-013, 2000LEN463, 2000LEN464 and 2000LEN471 were susceptible to Fusarium ox. f.sp. lentis with more than 50% mortality plants during two years. The accessions 302, 82-001, 91-001, 91-010, 91-017, 91-028, 93-024, 98-001, 2000LEN470 and 2000LEN472 were with plants mortality less than 50% during 2003 (Table. 1). Three accessions (302, 91-017, 91-028) were moderately susceptible with  results: 20%, 25% and 23% respectively. The remaining accessions from above group were susceptible with mortality plants between 25 and 50%. During 2004 the accessions 302, 91-001, 91-002, 91-028, 93-024, 98-001, 98-005 were moderately susceptible with results between 31.2% and 50% mortality plants at this stage (Table 2).  The only accessions were susceptible at this stage during all the experiment (2003-2004) 302, 91-001, 91-028, 93-024, 98-001. Four accessions (91-001, 91-028 and 98-001) were identifi ed as susceptible with total mortality plants till 50% during 2003 (Table 1). During the experiment in second year the same three accessions and 2000LEN466 124 Journal of Central European Agriculture Vol 7 (2006) No 1Tzvetelina Stoilova and Peter ChavdarovTable. 2  Resistance to Fusarium oxysporum f. sp. Lentis of Lentil genotypes, 2004 �.� Cat. � Sownseedsseedlings Mortality seedl.      Ns            % Mortality plants     ns.           % Total  mort. Pl.       Ns.                 % 1 2 3 4 5 6 7 8 9 101 302 20 12 8 40.0 5 40.7 13 65.0 2 345 20 2 18 90.0 2 100.0 20 100.0 3 82-001 20 5 15 75.0 4 80.0 19 95.0 4 82-003 20 8 12 60.0 5 62.5 17 85.0 5 91-001 20 15 5 25.0 5 33.3 10 50.0 6 91-002 20 14 6 30.0 7 50.0 13 65.0 7 91-006 20 0 20 100.0 - - 20 100.0 8 91-009 20 17 3 15.0 9 52.9 12 60.0 9 91-010 20 13 7 35.0 8 61.5 15 75.0 10 91-011 20 19 1 5.0 19 100.0 20 100.0 11 91-017 20 5 15 75.0 4 80.0 19 95.0 12 91-028 20 16 4 20.0 5 31.2 9 45.0 13 93-002 20 20 0 0.0 12 60.0 12 60.0 14 93-021 20 4 16 80.0 4 100.0 20 100.0 15 93-024 20 6 14 70.0 2 33.0 16 80.0 16 93-040 20 18 2 10.0 10 55.6 12 60.0 17 93-046 20 5 15 75.0 3 60.0 18 90.0 18 93-050 20 4 16 80.0 4 100.0 20 100.0 19 93-051 20 10 10 50.0 6 60.0 16 80.0 20 93-058 20 6 14 70.0 6 100.0 20 100.0 21 95-003 20 7 13 65.0 4 57.1 17 85.0 22 95-005 20 18 2 10.0 10 55.6 12 60.0 23 98-001 20 19 1 5.0 9 47.4 10 50.5 24 98-004 20 13 7 35.0 9 69.2 16 80.0 25 98-005 20 16 4 20.0 8 50.0 12 60.0 26 98-007 20 9 11 55.0 8 88.9 19 95.0 27 98-011 20 15 5 25.0 13 86.7 18 90.0 28 98-013 20 18 2 10.0 11 61.1 13 65.0 29 2000 LEN 463 20 12 8 40.0 8 66.7 16 80.0 30 2000 LEN 464 20 18 2 10.0 10 55.6 12 60.0 31 2000 LEN 465 20 0 20 100.0 - - 20 100.0 32 2000 LEN 466 20 19 1 5.0 9 47.4 10 50.0 33 2000 LEN 468 20 7 13 65.0 5 71.4 18 90.0 34  2000 LEN 470 20 15 5 25.0 8 53.3 13 65.0 35  2000 LEN 471 20 6 14 70.0 5 83.3 19 95.0 36 2000 LEN 472 20 14 6 30.0 10 71.4 16 80.0 were moderately susceptible. The total wilted plants were between 45% and 50%. The rest of the accessions which gave susceptible to highly susceptible reaction under glasshouse conditions showed variable reaction to Fusarium oxysporum f.sp. lentis during both years of the experiment. None of the studied accessions were immune or resistance to this pathogen under glasshouse conditions. CONCLUSIONS  The tested genotypes with different geographical origin were divided into three groups based on the symptoms observed, accessions. The fi rst group consisted accessions with highly susceptible to Fusarium oxysporum f. sp. lentis. This group is the biggest with reaction to Fusarium more than 50% wilted plants. The second and third were consisted from the accessions which gave MS to S reaction to Fusarium oxysporum f. sp. lentis under glasshouse conditions. They showed variable reaction during all the experimental period.Three of the studied accessions (91-001, 91-028 and 98-001) were susceptible with 45 and 50 % of total wilted plant.These genotypes could be used for developing cultivars especially for late-sown conditions.REFERENCES[1] Bayaa, B., Erskine W. and Khour, L., Arab Journal of Plant Protection, (1986) 4: 18-   119[2]  Bhatty, R.S., Compositionand quality of lentil (Lens culinaris Medic.): A review. Canadian Institute of Food Science and Technology,(1988), 21(2): 144 -160[3] Dobrev, D., Dissertation. (1987)[4 ] Erskine, W.; Bayaa, B. and Dolli, M., Arab Journal of Plant Protection (1990), 8: 34-37EVALUATION OF LENTIL GERMPLASM FOR DISEASE RESISTANCE TO FUSARIUM WILT (FUSARIUM OXYSPORUM F.SP. LENTIS)125J. Cent. Eur. Agric. (2006) 7:1, 121-126[5] Fleischmann, R. Pfl anzenbau, (1937), 14: 49-65[ 6 ] Hamdi A. and A.M.Hassanein, Survey of Fungal Diseases of Lentil in North Egypt. Lens Newsletter, (1996), 1&2, pp. 52-53[7] ICARDA. Food legume Improvement Program, Annual Report for 1990, Aleppo, Syria: ICARDA, (1990), 129 pp.[8] Kannaiyan, J. and Nene, Y.L., Indian Journal of Agricultural Science, (1976), 46: 165- 167[9] Khare, M. N, S.C.Agarwal and A.C. Jain.. Diseases of Lentil and their control.     Technical Bulletin JNKVV, Jabalpur, M.P., India, (1979) [10] Khare, M. N.. Wilt of Lentil,. Jabalpur,  M.P., India: JNKVV ,(1980), pp. 155 [11] Khare, M. N.. In: Lentils, (eds. C. Webb and G. Hawtin). UK: ICARDA/CAB. (1981), pp. 163-172[12] Mihov, M., I. Stoeva and P. Ivanov, Rastenievadni nauki, (1987), 24: 45-51[13] Saxena, D.R.and Khare, M.N., Indian Phytopathology (1988), 41:69-74[14] Stoilova T. and G. Pereira, Morphological characterization of 120 lentil (Lens culinaris Medic.) accessions. Lens Newsletter, (1999), 1&2, pp. 7-9[15] Toussoun,T.A. and Nelson, P.E., Fusarium: A Pictorial Guide to the Identifi cation of Fusarium Species,. University Park, Pennsylvania, USA: Pennsylvania State University press. (1976), pp. 43 [16] Waldia, S.R.; Singh, P.V. and Kharb, K.P.S., Stability of seed yield of some lentil genotypes in relation to seed size. Lens Newsletter, (1988), 15 (1): 17-22

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