白僵菌和乙酰甲胺磷对水稻抗氧化状态及土壤氮循环的影响

Effects of Beauveria bassiana and acephate on rice antioxidant status and soil nitrogen circulation

  • 摘要: 为了考察微生物农药白僵菌施用后在水稻系统中的生态安全性及其环境行为, 本文通过盆栽试验探讨了施加不同浓度白僵菌孢子悬液7.5×104 (孢子)·mL-1, 7.5×105 (孢子)·mL-1, 7.5×106 (孢子)·mL-1, 7.5×107 (孢子)·mL-1和化学农药乙酰甲胺磷对接种二化螟的水稻氧化还原状态及土壤氮循环相关的生物化学过程强度的影响。结果表明, 白僵菌处理对土壤氨化作用和硝化作用具有一定刺激作用, 最大提高幅度分别为12.4%和36.8%; 对反硝化作用表现为低浓度促进、高浓度抑制。乙酰甲胺磷处理在第10 d时对土壤氨化作用、硝化作用和反硝化作用的抑制率分别达到18.6%、45.3%和27.5%。乙酰甲胺磷处理后水稻AsA︰DHA(还原态抗坏血酸︰氧化态抗坏血酸)比值和GSH︰GSSG(还原态谷胱甘肽︰氧化态谷胱甘肽)比值降低, 并伴随抗氧化能力的降低。与此相反, 白僵菌处理能够提高AsA︰DHA和GSH︰GSSG。H2O2含量变化的结果表明二化螟接种和乙酰甲胺磷处理会导致过氧化程度升高, 而白僵菌的施加缓解了二化螟引起的过氧化状态。本试验使用荧光定量PCR对白僵菌的残留进行研究, 结果表明稻谷中并未有白僵菌残留, 白僵菌施用后对人类健康无害。上述结果表明, 与化学农药乙酰甲胺磷相比, 白僵菌是一种环境友好型的微生物农药, 其中7.5×104 (孢子)·mL-1的使用浓度效果较好。

     

    Abstract: Beauveria bassiana is a common entomopathogenic fungus with a global distribution. Isolates of B. bassiana can antagonize a variety of soil and plant pathogens. At present, studies have focused on B. bassiana insecticidal mechanism and symbiosis with plants. In order to investigate the ecological safety of B. bassiana and its environmental behavior in paddy systems, we conducted a potted trial to evaluate the effects of different concentrations of B. bassiana spore suspensions and chemical pesticide (acephate) on the redox state of rice and the intensity of biochemical processes related to soil nitrogen circulation. B. bassiana were expressed with the egfp gene (green fluorescent protein gene) through transformation with a vector, and real-time polymerase chain reaction (PCR) was used to detect B. bassiana in rice grains. To accomplish this, B. bassiana specific DNA primers (eGPF-F1/ eGFP-R1) were designed after the green fluorescent protein sequence marked B. bassiana. Amplification of B. bassiana DNA yielded a single 289 bp-long product using the eGPF-F1/eGFP-R1 primers. The detection limit was 10 fg·μL-1 of B. bassiana genomic DNA. The pot experiments consisted of seven treatments - CK (blank control group), CS (Chilo. suppressalis larvae), B1 (B. bassiana at 7.5×104 spores·mL-1 + CS), B10 (7.5×105 spores·mL-1 + CS), B100 (7.5×106 spores·mL-1 + CS), B1000 (7.5×107 spores·mL-1 + CS) and AE (acephate + CS). The results showed that B. bassiana stimulated the ammonification and nitrification of soils with maximum increases of 12.4% and 36.8%, respectively. Low concentration of B. bassiana facilitated denitrification, while high concentration inhibited denitrification. On the 10th day after treatment, acephate decreased ammonification, nitrification and denitrification rates by 18.6%, 45.3% and 27.5%, respectively. After inoculation with C. suppressalis and acephate application, ratios of AsA︰DHA and GSH︰GSSG reduced with decreasing antioxidant capacity. In contrast, B. bassiana treatment increased AsA︰DHA and GSH︰GSSG ratios. Changes in H2O2 content showed that C. suppressalis and acephate induced peroxidation, while B. bassiana reduced peroxidation intensity induced by C. suppressalis. Finally, B. Bassiana was not found in grains after RT-qPCR analysis. Overall, it was concluded that compared with chemical pesticide (acephate), B. bassiana was an effective and environment-friendly microbial pesticide.

     

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