Abstract
Fusarium spp. are phytopathogens causing fusarium head blight in wheat. They produce mycotoxins, mainly fumonisins, deoxynivalenol, and zearalenone. The study was conducted during two growing seasons (2020 and 2021) at the experimental field and laboratories of the Hungarian University of Agriculture and Life Sciences (MATE). The aim of the study was to determine the influence of growing season, nitrogen fertilisation, and wheat variety on Fusarium infection and mycotoxin production in wheat kernel. Zearalenone was not detected during the two growing seasons and deoxynivalenol was only detected in 2020. The results indicate that nitrogen fertilisation and wheat variety did not have statistically significant influence on Fusarium infection and mycotoxin production. The growing season had statistically significant influence on Fusarium infection and fumonisins production due to higher rainfall in 2021 compared to 2020 during the flowering period when the wheat spike is the most vulnerable to Fusarium infection.
1 Introduction
Wheat (Triticum aestivum L.) is one of the most cultivated crops around the world. It is grown across a wide range of environments. The primary use of wheat is for bread making. In addition, it is used in the production of bakery and confectionery products, animal feed, and ethanol. The genus Fusarium is a plant pathogen of wheat. It causes fusarium head blight (FHB), a major fungal disease in wheat production (Sifuentes dos Santos et al., 2013). Initial symptoms of FHB appear on the spike and grain. FHB reduces the quality of the grain and might decrease the yield up to 70%. Wheat is particularly susceptible to FHB infection during the period of anthesis and the early stages of grain development. Diseased grains are shrivelled, discoloured, and lightweight (Goswami and Kistler, 2004). Under favourable conditions, Fusarium species can produce mycotoxins, mainly deoxynivalenol (DON), zearalenone (ZEA), and fumonisins (FUM).
The presence of mycotoxins in food and feed can cause chronic or acute mycotoxicosis in animals and humans (Bottalico and Perrone, 2002). Deoxynivalenol (DON), commonly known as vomitoxin, causes food refusal, diarrhea, alimentary haemorrhaging, and contact dermatitis (Bennett and Klich, 2003). Zearalenone (ZEA) has estrogenic effects and reduces the reproductive capability of domestic animals (Biagi, 2009; Stanković et al., 2012). Fumonisins (FUM) are carcinogenic mycotoxins causing hepatotoxicity and apoptosis of the liver. In humans, it is linked with esophageal cancer (Marasas, 2001).
To minimise the risk of FHB and mycotoxins, some preventive measures should be applied to reduce their occurrence. The application of integral wheat protection measures such as cultivation of resistant cultivars, crop rotation, tillage, and application of appropriate fertilisers and fungicides can significantly reduce wheat infection by Fusarium species (Lemmens et al., 2004).
2 Materials and methods
The experiment was conducted during two growing seasons (2020 and 2021) at the experimental field and laboratories of the Hungarian University of Agriculture and Life Sciences (MATE), Crop Production Institute, Gödöllő, Hungary. The experimental site is in a hilly area with a close to average climatic zone of the country (47°35′40.8″N 19°22′08.4″E, 210 m above sea level).
The soil type of the experimental field is brown forest soil (Chromic Luvisol). Prior to sowing, the field was cleared, ploughed, rotor-tilled, and the seedbed was prepared. The plots were sown and harvested with plot machines. The trial design was that of a split-plot with main plots consisting of different wheat varieties and subplots consisting of different nitrogen doses. Main plots and subplots were 50 cm apart horizontally and 30 cm apart vertically, and the area of each subplot was 5 m2. Each treatment had three replications. The wheat varieties used were: Alföld, Mv Kolompos, and Mv Karéj. Nitrogen fertiliser (NH4NO3) was applied twice during the growing season, the first application was done at tillering stage and the second application at heading stage. The doses of nitrogen in the first application were: 40, 80, and 120 kg N ha−1. In the second application 40 kg N ha−1 was added only. Plots without nitrogen topdressing were used as control. Fusarium percentage was calculated by counting the number of colonies that formed on wheat kernels disinfected with a solution of pentachloronitrobenzene (PCNB) and chloramphenicol (100 kernels from each treatment) incubated for 7 days under laboratory conditions on Nash and Snider Fusarium selective medium (distilled water 1 L, peptone 15 g, KH2PO4 1 g, MgSO47H2O 0.5 g, agar 20 g, PCNB 1 g, chloramphenicol 100 ppm). Mycotoxin concentrations of deoxynivalenol (DON), zearalenone (ZEA), and fumonisins (FUM) were analysed using ROSA FAST 5 Quantitative Test by Charm Sciences.
For the statistical evaluation of the results, analysis of variance (ANOVA) module of the IBM SPSS V.21 software at 5% significance level with subsequent Tukey's test was performed to determine the influence of growing season, nitrogen fertilisation and wheat variety on Fusarium infection and mycotoxin production in wheat kernel.
3 Results and discussion
The study of the influence of growing season, wheat variety, and nitrogen fertilisation on Fusarium infection and subsequent mycotoxin production in wheat kernel was carried out in 2020 and 2021. The growing season significantly affected Fusarium infection (F = 187.31, P = 0.000) and fumonisins concentration (F = 4.7, P = 0.03) but did not significantly affect deoxynivalenol concentration (F = 3.61, P = 0.06) (Figs 1 and 2, Table 2). Fusarium infection was higher in 2021 (93.1%) than in 2020 (46.9%) (Fig. 1). Zearalenone was not detected throughout the two growing seasons. Fumonisins concentration (total mean = 22.2 ppb) was higher than that of deoxynivalenol (total mean = 15.97 ppb) (Table 1). Deoxynivalenol was not detected in 2021, its concentration was 31.9 ppb in 2020 (Fig. 2). Fumonisins concentration was higher in 2021 (30.6 ppb) than in 2020 (13.9 ppb) (Fig. 2). Rainfall (mm) measurements were collected from the Hungarian National Meteorological Service during the flowering period (May) when wheat is most susceptible to Fusarium infection. Rainfall during the flowering period (May) in 2021 was 123.1 mm, higher than in 2020 (39.8 mm), this increase in rainfall could explain the increased Fusarium percentage and fumonisins concentration.
Effect of growing season on Fusarium percentage (%)
Citation: Acta Alimentaria 51, 2; 10.1556/066.2022.00036
Effect of growing season on mycotoxin concentration (ppb)
Citation: Acta Alimentaria 51, 2; 10.1556/066.2022.00036
Descriptive statistics of Fusarium percentage (%), DON and FUM concentration (ppb) affected by growing season, wheat variety, and nitrogen dosage (kg N ha−1)
| Mean | Std. deviation | Std. error | Minimum | Maximum | |||
| Growing season | Fusarium | 2020 | 46.86 | 19.31 | 3.22 | 12 | 90 |
| 2021 | 93.06 | 6.11 | 1.02 | 76 | 100 | ||
| Total | 69.96 | 27.26 | 3.21 | 12 | 100 | ||
| DON | 2020 | 31.94 | 100.82 | 16.8 | 0 | 500 | |
| 2021 | 0 | 0 | 0 | 0 | 0 | ||
| Total | 15.97 | 72.59 | 8.55 | 0 | 500 | ||
| FUM | 2020 | 13.89 | 22.71 | 3.79 | 0 | 50 | |
| 2021 | 30.56 | 40.14 | 6.69 | 0 | 200 | ||
| Total | 22.22 | 33.45 | 3.94 | 0 | 200 | ||
| Wheat variety | Fusarium | Alföld | 64.88 | 30.27 | 6.18 | 22 | 100 |
| Kolompos | 72.00 | 25.19 | 5.14 | 22 | 100 | ||
| Karéj | 73.00 | 26.50 | 5.41 | 12 | 100 | ||
| Total | 69.96 | 27.26 | 3.21 | 12 | 100 | ||
| DON | Alföld | 25.00 | 103.21 | 21.07 | 0 | 500 | |
| Kolompos | 8.33 | 40.82 | 8.33 | 0 | 200 | ||
| Karéj | 14.58 | 61.64 | 12.58 | 0 | 300 | ||
| Total | 15.97 | 72.59 | 8.55 | 0 | 500 | ||
| FUM | Alföld | 22.92 | 25.45 | 5.19 | 0 | 50 | |
| Kolompos | 12.50 | 22.12 | 4.51 | 0 | 50 | ||
| Karéj | 31.25 | 46.19 | 9.43 | 0 | 200 | ||
| Total | 22.22 | 33.45 | 3.94 | 0 | 200 | ||
| Nitrogen dosage | Fusarium | 0 | 69.39 | 28.17 | 6.64 | 12 | 100 |
| 40+40 | 70.33 | 29.21 | 6.89 | 22 | 100 | ||
| 80+40 | 66.44 | 28.54 | 6.73 | 22 | 100 | ||
| 120+40 | 73.67 | 24.79 | 5.84 | 24 | 96 | ||
| Total | 69.96 | 27.26 | 3.21 | 12 | 100 | ||
| DON | 0 | 27.78 | 117.85 | 27.78 | 0 | 500 | |
| 40+40 | 16.67 | 70.71 | 16.67 | 0 | 300 | ||
| 80+40 | 16.67 | 51.45 | 12.13 | 0 | 200 | ||
| 120+40 | 2.78 | 11.79 | 2.78 | 0 | 50 | ||
| Total | 15.97 | 72.59 | 8.55 | 0 | 500 | ||
| FUM | 0 | 16.67 | 29.70 | 7.00 | 0 | 100 | |
| 40+40 | 19.44 | 25.08 | 5.91 | 0 | 50 | ||
| 80+40 | 33.33 | 48.51 | 11.43 | 0 | 200 | ||
| 120+40 | 19.44 | 25.08 | 5.91 | 0 | 50 | ||
| Total | 22.22 | 33.45 | 3.94 | 0 | 200 |
0: no nitrogen application.
40 + 40: the first nitrogen application was 40 kg N ha−1 and the second was 40 kg N ha−1.
80 + 40: the first nitrogen application was 80 kg N ha−1 and the second was 40 kg N ha−1.
120 + 40: the first nitrogen application was 120 kg N ha−1 and the second was 40 kg N ha−1.
The wheat variety did not significantly affect Fusarium infection (F = 0.63, P = 0.54) and subsequent mycotoxin production (DON, F = 0.32, P = 0.73; FUM, F = 1.94 P = 0.15) (Table 2).
Analysis of variance for Fusarium percentage and DON, FUM concentrations affected by growing season, wheat variety, and nitrogen dosage
| Source of variation | Sum of Squares | df | Mean Square | F | Sig. | ||
| Growing season | Fusarium | Between Groups | 38410.68 | 1 | 38410.68 | 187.31 | 0.00 |
| Within Groups | 14354.19 | 70 | 205.06 | ||||
| Total | 52764.88 | 71 | |||||
| DON | Between Groups | 18368.06 | 1 | 18368.06 | 3.61 | 0.06 | |
| Within Groups | 355763.89 | 70 | 5082.34 | ||||
| Total | 374131.94 | 71 | |||||
| FUM | Between Groups | 5000.00 | 1 | 5000.00 | 4.7 | 0.03 | |
| Within Groups | 74444.44 | 70 | 1063.49 | ||||
| Total | 79444.44 | 71 | |||||
| Wheat variety | Fusarium | Between Groups | 942.250 | 2 | 471.13 | 0.63 | 0.54 |
| Within Groups | 51822.625 | 69 | 751.05 | ||||
| Total | 52764.875 | 71 | |||||
| DON | Between Groups | 3402.778 | 2 | 1701.39 | 0.32 | 0.73 | |
| Within Groups | 370729.167 | 69 | 5372.89 | ||||
| Total | 374131.944 | 71 | |||||
| FUM | Between Groups | 4236.111 | 2 | 2118.06 | 1.94 | 0.15 | |
| Within Groups | 75208.333 | 69 | 1089.98 | ||||
| Total | 79444.444 | 71 | |||||
| Nitrogen dosage | Fusarium | Between Groups | 478.15 | 3 | 159.38 | 0.21 | 0.89 |
| Within Groups | 52286.72 | 68 | 768.92 | ||||
| Total | 52764.88 | 71 | |||||
| DON | Between Groups | 5659.72 | 3 | 1886.57 | 0.35 | 0.79 | |
| Within Groups | 368472.22 | 68 | 5418.71 | ||||
| Total | 374131.94 | 71 | |||||
| FUM | Between Groups | 3055.56 | 3 | 1018.52 | 0.91 | 0.44 | |
| Within Groups | 76388.89 | 68 | 1123.37 | ||||
| Total | 79444.44 | 71 |
df: degree of freedom; Sig.: significance; Significance level = P < 0.05.
The nitrogen fertilisation did not significantly affect Fusarium infection (F = 0.21, P = 0.89) and subsequent mycotoxin production (DON, F = 0.35, P = 0.79; FUM, F = 0.91, P = 0.44) (Table 2).
In our study, the different climatic conditions that prevailed during 2020/2021 could be the reason for the increase in Fusarium percentage and fumonisins concentration. According to Brennan et al. (2003), the development of FHB in wheat depends on rainfall. Bryła et al. (2016) also stated that the development, growth, and spread of Fusarium fungi and the degree of infection strongly depend on rainfall. Mesterházy et al. (1999) pointed out that climatic conditions may play an important role in Fusarium infection of wheat. González et al. (2008) suggested that the relatively high level of natural Fusarium contamination could be due to a high rainfall period that occurred during the flowering stage. Risk of FHB in wheat plants depends also on genetically determined resistance of the given wheat cultivar to Fusarium spp. (Zhang et al., 2008). According to these authors, the main factors affecting Fusarium contamination of wheat were weather conditions and susceptibility of wheat cultivars to Fusarium spp.
In our study, nitrogen dosage did not influence Fusarium contamination and mycotoxin production. Krnjaja et al. (2015) found that nitrogen fertilisation did not increase FHB intensity. Kuzdraliński et al. (2014) reported that the rate of autumn N fertilisation did not affect the number of Fusarium detections.
Bernhoft et al. (2012) concluded that farming system (organic versus conventional) impacted Fusarium infestation, and that organic management tended to reduce Fusarium contamination and mycotoxins. However, Fusarium infestation and mycotoxin concentrations may be influenced by a range of factors such as local topography and local climate. Oldenburg et al. (2007) concluded that nitrogen rates of up to 240 kg N ha−1 did not influence Fusarium growth and their production of mycotoxins in wheat grains. According to Parry et al. (1995), the impact of nitrogen fertilisation on Fusarium infestation remains unclear. Moreover, Aufhammer et al. (2000) concluded that nitrogen fertilisation did not stimulate Fusarium infection and mycotoxin production. In addition, Martin et al. (1991) observed that nitrogen rates increasing from 70 to 170 kg N ha−1 significantly increased the occurrence of Fusarium infected grains in wheat. According to Lemmens et al. (2004), increasing nitrogen fertilisation rates up to 80 kg N ha−1 significantly affected Fusarium infection and subsequent mycotoxin contamination in wheat. However, Lemmens et al. (2004) concluded that the occurrence of Fusarium spp. could not be solely based on the nitrogen input in crop production. All these results suggest that the effect of nitrogen fertilisation can only partially influence the creation of favourable conditions for the occurrence of Fusarium spp.
4 Conclusions
Growing season, nitrogen fertilisation and wheat variety were studied to evaluate Fusarium infection and mycotoxin production in wheat kernel. The results indicate that nitrogen fertilisation and wheat variety did not show statistically significant influence on Fusarium infection and mycotoxin production. The growing season showed statistically significant influence on Fusarium infection and fumonisins production due to higher rainfall in 2021 compared to 2020 during the flowering period when the wheat spike is the most vulnerable to Fusarium infection.
Acknowledgment
This research was supported by the Doctoral School of Plant Science of the Hungarian University of Agriculture and Life Sciences. The PhD students involved were sponsored by the Stipendium Hungaricum scholarship. The authors would like to express thanks to all colleagues and technical staff on-site and in laboratories for their assistance and valuable contribution to the implementation of this study.
References
Aufhammer, W. , Kübler, E. , Kaul, H.P. , Hermann, W. , Hohn, D. , and Yi, C. (2000). Infection with head blight (F. graminearum, F. culmorum) and deoxynivalenol concentration in winter wheat as influenced by N fertilization. Pflanzenbauwissenschaften, 4: 72–78.
Bennett, J.W. and Klich, M. (2003). Mycotoxins. Clinical Microbiology Reviews, 16: 497–516. https://doi.org/10.1128/CMR.16.3.497-516.2003.
Bernhoft, A. , Torp, M. , Clasen, P.E. , Løes, A.K. , and Kristoffersen, A.B. (2012). Influence of agronomic and climatic factors on Fusarium infestation and mycotoxin contamination of cereals in Norway. Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 29: 1129–1140. https://doi.org/10.1080/19440049.2012.672476.
Biagi, G. (2009). Dietary supplements for the reduction of mycotoxin intestinal absorption in pigs. Biotechnology in Animal Husbandry, 25: 539–546. https://doi.org/10.2298/BAH0906539B.
Bottalico, A. and Perrone, G. (2002). Toxigenic Fusarium species and mycotoxins associated with head blight in small-grain cereals in Europe. European Journal of Plant Pathology, 108: 611–624. https://doi.org/10.1023/A:1020635214971.
Brennan, J.M. , Fagan, B. , van Maanen, A. , Cooke, B.M. , and Doohan, F.M. (2003). Studies on in vitro growth and pathogenicity of European Fusarium fungi. European Journal of Plant Pathology, 109: 577–587. https://doi.org/10.1023/A:1024712415326.
Bryła, M. , Waśkiewicz, A. , Podolska, G. , Szymczyk, K. , Jędrzejczak, R. , Damaziak, K. , and Sułek, A. (2016). Occurrence of 26 mycotoxins in the grain of cereals cultivated in Poland. Toxins, 8(6): 160. https://doi.org/10.3390/toxins8060160.
González, H.H.L. , Moltó, G.A. , Pacin, A. , Resnik, S.L. , Zelaya, M.J. , Masana, M. , and Martínez, E.J. (2008). Trichothecenes and mycoflora in wheat harvested in nine locations in Buenos Aires province, Argentina. Mycopathologia, 165: 105–114. https://doi.org/10.1007/S11046-007-9084-X.
Goswami, R.S. and Kistler, H.C. (2004). Heading for disaster: Fusarium graminearum on cereal crops. Molecular Plant Pathology, 5(6): 515–525. https://doi.org/10.1111/J.1364-3703.2004.00252.X.
Krnjaja, V. , Mandić, V. , Lević, J. , Stanković, S. , Petrović, T. , Vasić, T. , and Obradović, A. (2015). Influence of N-fertilization on Fusarium head blight and mycotoxin levels in winter wheat. Crop Protection, 67: 251–256. https://doi.org/10.1016/J.CROPRO.2014.11.001.
Kuzdraliński, A. , Szczerba, H. , Tofil, K. , Filipiak, A. , Garbarczyk, E. , Dziadko, P. , Muszyńska, M. , and Solarska, E. (2014). Early PCR-based detection of Fusarium culmorum, F. graminearum, F. sporotrichioides and F. poae on stem bases of winter wheat throughout Poland. European Journal of Plant Pathology, 140: 491–502. https://doi.org/10.1007/s10658-014-0483-9.
Lemmens, M. , Haim, K. , Lew, H. , and Ruckenbauer, P. (2004). The effect of nitrogen fertilization on Fusarium head blight development and deoxynivalenol contamination in wheat. Journal of Phytopathology, 152(1): 1–8. https://doi.org/10.1046/j.1439-0434.2003.00791.x.
Marasas, W.F.O. , Miller, J.D. , Riley, R.T. , and Visconti, A. (2001). Fumonisins occurrence, toxicology, metabolism and risk assessment, In: Summerell, B.A. , Leslie, J.F. , Backhouse, D. , Bryden, W.L. , and Burgess, L.W. (Eds.), Fusarium. Paul E. Nelson memorial symposium. APS Press, St. Paul, Minn. pp. 332–359.
Martin, R.A. , MacLeod, J.A. , and Caldwell, C. (1991). Influences of production inputs on incidence of infection by Fusarium species on cereal seed. Plant Discipline, 75: 784–788.
Mesterházy, Á. , Bartók, T. , Mirocha, C.G. , and Komoróczy, R. (1999). Nature of wheat resistance to Fusarium head blight and the role of deoxynivalenol for breeding. Plant Breeding, 118: 97–110. https://doi.org/10.1046/J.1439-0523.1999.118002097.X.
Oldenburg, E. , Bramm, A. , and Valenta, H. (2007). Influence of nitrogen fertilization on deoxynivalenol contamination of winter wheat - experimental field trials and evaluation of analytical methods. Mycotoxin research, 23: 7–12. https://doi.org/10.1007/BF02946018.
Parry, D.W. , Jenkinson, P. , and McLeod, L. (1995). Fusarium spike blight (scab) in small grain cereals review. Plant Pathology, 44(2): 207–238. https://doi.org/10.1111/J.1365-3059.1995.TB02773.X.
Sifuentes dos Santos, J. , Souza, T.M. , Ono, E.Y.S. , Hashimoto, E.H. , Bassoi, M.C. , Miranda, M.Z. de , Itano, E.N. , Kawamura, O. , and Hirooka, E.Y. (2013). Natural occurrence of deoxynivalenol in wheat from Paraná State, Brazil and estimated daily intake by wheat products. Food Chemistry, 138: 90–95. https://doi.org/10.1016/j.foodchem.2012.09.100.
Stanković, S. , Lević, J. , Ivanović, D. , Krnjaja, V. , Stanković, G. , and Tančić, S. (2012). Fumonisin B1 and its co-occurrence with other fusariotoxins in naturally contaminated wheat grain. Food Control, 23: 384–388. https://doi.org/10.1016/J.FOODCONT.2011.08.003.
Zhang, J.X. , Jin, Y. , Rudd, J.C. , and Bockelman, H.E. (2008). New fusarium head blight resistant spring wheat germplasm identified in the USDA National Small Grains Collection. Crop Science, 48: 223–235. https://doi.org/10.2135/cropsci2007.02.0116.
