柴喜存, 周天宇, 马海霞, 王世鹏, 刘玉涛. 缺氮处理禾谷镰刀菌在氮素恢复后的致病力变化[J]. 农业工程学报, 2021, 37(4): 175-180. DOI: 10.11975/j.issn.1002-6819.2021.4.021
    引用本文: 柴喜存, 周天宇, 马海霞, 王世鹏, 刘玉涛. 缺氮处理禾谷镰刀菌在氮素恢复后的致病力变化[J]. 农业工程学报, 2021, 37(4): 175-180. DOI: 10.11975/j.issn.1002-6819.2021.4.021
    Chai Xicun, Zhou Tianyu, Ma Haixia, Wang Shipeng, Liu Yutao. Pathogenicity variation of Fusarium graminearum suffered nitrogen deficiency after nitrogen recovery[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2021, 37(4): 175-180. DOI: 10.11975/j.issn.1002-6819.2021.4.021
    Citation: Chai Xicun, Zhou Tianyu, Ma Haixia, Wang Shipeng, Liu Yutao. Pathogenicity variation of Fusarium graminearum suffered nitrogen deficiency after nitrogen recovery[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2021, 37(4): 175-180. DOI: 10.11975/j.issn.1002-6819.2021.4.021

    缺氮处理禾谷镰刀菌在氮素恢复后的致病力变化

    Pathogenicity variation of Fusarium graminearum suffered nitrogen deficiency after nitrogen recovery

    • 摘要: 病害高发是限制秸秆还田技术实施的因素之一。为探讨因秸秆还田导致的土壤中氮素水平的变化对植物病原菌致病力的影响,该研究选择常见的随麦秸秆还田携入土中的禾谷镰刀菌为研究对象,通过室内模拟分析5种不同缺氮处理(氮素占比分别为0、25%、50%、75%和100%,其中100% 表示不缺氮处理,培养基中氯化铵浓度为1.0 g/L)下禾谷镰刀菌进行氮素恢复后生长量、孢子萌发率、毒素产量和侵染长度等的变化。结果发现,不同处理禾谷镰刀菌的生长速率具有明显差别,完全缺氮下(氮素占比为0)禾谷镰刀菌在氮素恢复24 h时,单位体积菌丝质量达到0.354 g/mL,明显高于其他处理(P<0.05),氮素恢复168 h后其孢子产量仅为8.2×105个/mL,显著低于其他处理(P<0.05),但产生的分生孢子在氮素恢复后具有最高的萌发率,达到46.00%,显著高于其他处理(P<0.05)。在氮素占比仅为25%处理中,当氮素恢复24 h禾谷镰刀菌的毒素产量会显著高于其他处理(P<0.05),达到了1.361 μg/g;在不缺氮及氮素占比为50%和75%时,氮素恢复72 h禾谷镰刀菌的毒素产量才显著高于其他处理(P<0.05);在氮素恢复培养过程中,禾谷镰刀菌的侵染能力呈现出先下降后上升的趋势,氮素恢复72 h时,完全缺氮处理小麦苗病变长度为7.73 mm,显著低于氮素占比25%、50%和100%处理(P<0.05)。结果表明,短期缺氮(168 h)后补给氮素能够消除氮缺乏对禾谷镰刀菌生长造成的影响;在氮素恢复初期(2 h)禾谷镰刀菌致病能力会有所下降,随着氮素的消耗,禾谷镰刀菌处于部分缺氮环境时的产毒和侵染能力又大幅度提升,这将为秸秆还田后合理施肥策略的制定提供理论参考。

       

      Abstract: Abstract: To clarify the effects of nitrogen recovery on the pathogenicity variation of Fusarium graminearum (F. graminearum) suffered nitrogen starvation, an off-line simulation method was used to test the growth, spore germination, toxicity and infectivity of F. graminearum. The MM-N media containing 0, 25%, 50%, 75% and 100% nitrogen were set as five treatments, among which 100% nitrogen level treatment contained 1.0 g/L ammonium chloride. Hyphae growing in Potato Dextrose Agar liquid media (PDA) for 72 h were then shifted to MM-N media for the nitrogen starvation. After cultured in different nitrogen levels media for 168 h, hyphae were shifted to MM total nitrogen media for the nitrogen recovery. Cultures were sampled at 0, 2, 24, 48 and 72 h for further analysis. Five millilitres of culture were sampled and centrifuged at 10 000 rpm for 10 min to separate the mycelium from the media; then the weights were recorded for charactering the growth of F. graminearum. The spores of F. graminearum were calculated and then transferred to MM total nitrogen media; after cultured for 48 h, the germination of spores in 40 μL cultures was observed under a microscope and the germination rate was calculated. Toxin Deoxynivalenol (DON) was extracted using specific immunoaffinity columns; the samples were evaporated under a low nitrogen stream at 50°C and resuspended in the mobile phase (methanol: water=18:82, v/v). The determination of DON was performed using high-performance liquid chromatography with Shimadzu SPD-M20A Infinity II Diode Array Detector (DAD); A 150 mm×4.6 mm C18 column with a particle size of 0.45 μm from Phenomenex was used; The mobile phase comprised methanol and water (18:82, v/v) and was set at a flow rate of 0.8 mL/min; The column temperature was 35°C, the injection volume was 20 μL, and the total run time was 15 min for mycotoxin analyses. The infection lengths of wheat seedlings were used to test the infectivity of F. graminearum, and the lesion length was measured with a digital calliper. Significant differences were not found for the growth rate of F. graminearum treated by different nitrogen levels deficiency after nitrogen recovery for 24 h (P<0.05). The biomass of total nitrogen deficiency treatment was significantly higher than the other treatments (P<0.05), reaching 0.354 g/mL liquid media. The biomass of each treatment was not significantly different after 48 h culture. However, after 72 h of nitrogen recovery, the biomass of nitrogen treatment (25%) was significantly higher than that of the other treatments (P<0.05). The treatment of total nitrogen deficiency only produced 8.2×105 spores/mL, but it had the highest spore germination rate, reaching 46.00%. The DON production of F. graminearum of total nitrogen deficiency treatment was always low after the nitrogen recovery. The nitrogen treatment (25%) for 24 h and the nitrogen proportion 50%, 75% and 100% treatments for 72 h had the higher toxin production compared to the other treatments at the same time (P<0.05), the toxin content were 1.361, 0.515,1.409 and 0.540 μg/g. After the nitrogen recovery, the pathogenicity of F. graminearum showed a trend of decreasing firstly and then increasing. At the initial stage of nitrogen recovery, the infection length of nitrogen proportion 100% treatment reached 13.28 mm. Then, the infection length was decreased significantly from 2 h to 48 h, and increased to 11.96 mm at 72 h. The infection lengths of the three treatments (nitrogen proportion 25%, 50% and 75%), reached the lowest at 24 h, and then increased to the maximum at 72 h after nitrogen recovery. Cultured for 72 h, the infection length of the treatment with total nitrogen starvation was 7.73 mm, significantly lower than the treatments of nitrogen proportion of 25%, 50% and 100% (P<0.05). The results would provide valuable information for establishing rational strategy of fertilizer application after straw incorporation into soils.

       

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