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用于T6对线虫生物防治潜力的发酵培养基及参数的优化

Optimization of the Fermentation Media and Parameters for the Bio-control Potential of T6 Against Nematodes.

作者信息

Zhang Shuwu, Gan Yantai, Liu Jia, Zhou Jingjiang, Xu Bingliang

机构信息

Gansu Provincial Key Laboratory of Arid Land Crop Science, Gansu Agricultural University, Lanzhou, China.

College of Plant Protection, Gansu Agricultural University, Lanzhou, China.

出版信息

Front Microbiol. 2020 Sep 30;11:574601. doi: 10.3389/fmicb.2020.574601. eCollection 2020.

Abstract

The cereal cyst nematode is one of the important soil-borne pathogens of cereal crops and causes high yield losses worldwide. spp. formulations are applied as commercial bio-control agents against soil-borne plant pathogens such as . However, the relationship between fermentation parameters and its bio-control potential against has not been exclusively established. In the present study, the effect of 10 different fermentation media and conditions on the nematicidal activity of T6 (T6) was evaluated with a single-factor method and a Plackett-Burman design, and the interaction between different fermentation parameters was investigated by a Box-Behnken design. The variables for enhancing the nematicidal activity of T6 culture filtrates were explored and optimized using response surface methodology (RSM). The Minor Medium (MM) plus wheat bran-2 medium was found to be the most effective fermentation medium for T6 culture filtrates against the second stage juveniles (J2s) of . The maximum mortality of the J2s was obtained using the T6 culture filtrates under the following fermentation conditions: initial pH 6, 28°C culture temperature, 180 rpm rotating speed, 60 ml of fermentation media, 7 days of incubation time, and 1 ml of inoculation volumes. Among these parameters, the initial pH, inoculation volume, and incubation day were identified as the most significant parameters and critical independent variables for enhancing the nematicidal activity of the T6 culture filtrates. After further optimizations based on statistical predictions, the highest nematicidal activity (92.42%) was obtained with the T6 culture filtrates fermented under an initial pH of 6.06, an inoculation volume of 1.62 ml, and an incubation time of 7.15 days. The nematicidal activity was increased approximately by as high as 1.07% compared with that before optimization. Bio-control efficacy of T6 culture filtrates was 83.88% at the 70 day after wheat seeds sowing in greenhouse experiments. The results from the validation experiments agreed with the model predictions. Our study has improved the bio-control potential of spp. against the plant-parasitic nematodes and provided a cost-efficient bio-resource in the future development of novel bio-control agents.

摘要

谷物孢囊线虫是谷物作物重要的土传病原菌之一,在全球范围内造成高额产量损失。 spp. 制剂作为商业生物防治剂用于防治诸如 等土传植物病原菌。然而, 的发酵参数与其对 的生物防治潜力之间的关系尚未完全确立。在本研究中,采用单因素法和Plackett-Burman设计评估了10种不同发酵培养基和条件对 T6(T6)杀线虫活性的影响,并通过Box-Behnken设计研究了不同发酵参数之间的相互作用。使用响应面方法(RSM)探索并优化了提高T6培养滤液杀线虫活性的变量。发现Minor培养基(MM)加麦麸-2培养基是T6培养滤液针对 第二阶段幼虫(J2s)最有效的发酵培养基。在以下发酵条件下使用T6培养滤液可使J2s的死亡率达到最高:初始pH 6、培养温度28°C、转速180 rpm、发酵培养基60 ml、培养时间7天以及接种量1 ml。在这些参数中,初始pH、接种量和培养天数被确定为提高T6培养滤液杀线虫活性的最显著参数和关键自变量。基于统计预测进行进一步优化后,在初始pH 6.06、接种量1.62 ml和培养时间7.15天的条件下发酵得到的T6培养滤液具有最高的杀线虫活性(92.42%)。与优化前相比,杀线虫活性提高了约1.07%。在温室试验中,小麦种子播种70天后,T6培养滤液的生物防治效果为83.88%。验证实验结果与模型预测相符。我们的研究提高了 spp. 对植物寄生线虫的生物防治潜力,并为新型生物防治剂的未来开发提供了一种经济高效的生物资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ca/7554348/dc7e9f8d2f5b/fmicb-11-574601-g001.jpg

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