菌糠水提液对马铃薯致病疫霉的抑制机理

doi:10.13304/j.nykjdb.2023.0159

摘要/Abstract

摘要：

菌糠是菌菇生产后残留的物质，含有丰富的无机盐和有机质等成分。利用热水浸提法制备香菇菌糠水提液（water extract from spent mushroom substrate，WESMS），通过紫外分光光度计和Zeta电位及粒径仪对其紫外吸收情况、表面电荷和水中分散粒径进行表征，采用平板渗透法及十字交叉测量直径法计算WESMS对致病疫霉的抑制率，利用光学显微镜和扫描电镜观察病原菌细胞形态的损伤程度，并采用琼脂糖凝胶电泳法分析WESMS对致病疫霉 DNA的影响。结果表明，WESMS在紫外线A（UVA，315~400 nm）、B（UVB，280~315 nm）和C（UVC，100~280 nm）波段均有吸收，在水中的分散粒径为3 649.27 nm。抑菌试验表明，WESMS对病原菌的生长具有抑制作用，随着提取液体积百分浓度的增加，抑制效果更明显，在高体积百分浓度（6.25% WESMS）作用下对病原菌的抑制率近100%。经WESMS处理后病原菌的菌丝更加扭曲、扁平，且褶皱明显增多，表明WESMS可对细胞造成明显破坏；WESMS处理组的DNA条带亮度暗于对照组，损伤程度与WESMS体积百分浓度呈正相关。研究结果为菌糠的资源化利用及其对马铃薯晚疫病的有效防治提供科学依据和技术支撑。

关键词: 香菇菌糠, 马铃薯晚疫病, 致病疫霉, 抗菌机理

Abstract:

Mushroom substrate is the residue of mushroom production， which is rich in inorganic salts and organic matter. The water extract from spent mushroom substrate（WESMS） of Lentinula edodes was prepared by hot water extraction，the ultraviolet （UV） absorption， surface charge and particle size distribution were characterized using an UV spectrophotometer， Zeta potential and particle size analyzer. The inhibition rate of WESMS against Phytophthora infestans was calculated by plate penetration and cross diameter measurement. Further， both optical microscopy and scanning electron microscopy were used to observe the damage caused by WESMS to the morphology of the pathogen cells. Subsequently， the impact of WESMS on the DNA of Phytophthora infestans wasanalyzed by agarose gel electrophoresis. The results showed that WESMS could absorb UV rays at A （UVA， 315~400 nm）， B （UVB， 280~315 nm） and C （UVC， 100~280 nm） bands. Moreover， the particle size distribution in water was 3 649.27 nm. The antibacterial experiment showed that WESMS had an inhibitory effect on the growth of pathogens. The inhibitory effect became more obvious with the increase of the volume percentage concentration of WESMS. The inhibition rate of WESMS against pathogens was nearly 100% at a high volume concentration of 6.25%. The mycelia of pathogenic cells treated with WESMS exhibited obvious distortion， flatness and wrinkles compared with the control group， which indicated that WESMS could cause severe damage to cells. In addition， the DNA band brightness of the treatment group was darker than that of the control group， and the degree of DNA damage was positively related to the volume percentage concentration of WESMS. Above results provided scientific basis and technical support for the utilization of spent mushroom substrate and its use in the effective control of potato late blight.

Key words: spent mushroom substrate of Lentinula edodes, potato late blight, Phytophthora infestans, antimicrobial mechanism

中图分类号:

S435.32

莫雯婧, 陈洪森, 桂芳泽, 洪慈清, 蔡鑫铠, 关雄, 潘晓鸿. 菌糠水提液对马铃薯致病疫霉的抑制机理[J]. 中国农业科技导报, 2024, 26(5): 129-137.

Wenjing MO, Hongsen CHEN, Fangze GUI, Ciqing HONG, Xinkai CAI, Xiong GUAN, Xiaohong PAN. Inhibition Mechanism of Water Extract from Spent Mushroom Substrateagainst Phytophthora infestans in potatoes[J]. Journal of Agricultural Science and Technology, 2024, 26(5): 129-137.

图/表 7

图1 香菇菌糠提取液的制备及表征A. 香菇菌糠提取液制备流程； B. 香菇菌糠提取液紫外吸收可见光谱

Fig. 1 Preparation and characterization of WESMSA. Preparation process of WESMS； B. UV-vis spectrum of WESMS

图2 马铃薯致病疫霉与不同体积百分浓度的香菇菌糠水提液作用后的平板

Fig. 2 Plate of Phytophthora infestans in potato after treated with different volume percentage concentrations of WESMS

表1 不同体积百分浓度的香菇菌糠水提液对马铃薯致病疫霉菌的抑菌率

Table 1 Inhibitory rate of WESMS with different volume percentage concentrations against Phytophthora infestans in potato

时间 Time/d	抑制率 Inhibition rate/%
时间 Time/d	0.33% WESMS	0.66% WESMS	1.64% WESMS	3.23% WESMS	6.25% WESMS
5	16.67±2.62 d	28.79±4.73 c	56.06±8.61 b	94.70±5.72 a	100.00±0.00 a
7	17.29±2.53 e	23.54±0.36 d	45.42±1.91 c	81.25±3.31 b	99.79±0.36 a
9	4.13±0.00 e	18.32±8.89 d	53.99±6.79 c	82.78±5.48 b	99.17±0.00 a
11	2.00±0.00 d	6.44±2.70 d	51.45±2.55 c	73.33±6.96 b	96.67±0.00 a

图3 光学显微镜下（40×）的菌丝形态A~C：空白对照组菌丝，分别为与平板中心点距离不同处采集的菌丝；D~F：6.25%体积百分浓度WESMS处理的菌丝

Fig. 3 Mycelial morphology under optical microscope （40×）A~C： Mycelium of blank control group， the mycelia are collected at different distances from the center point of the plate； D~F： Mycelium treated with 6.25% volume percentage concentration of WESMS

图4 扫描电镜下菌丝形态A~D：空白对照组菌丝； E~H：6.25%体积百分浓度WESMS处理的菌丝

Fig. 4 Mycelial morphology under scanning electron microscopeA~D： Mycelium of blank control group； E~H： Mycelium treated with 6.25% volume percentage concentration of WESMS

图5 卵菌基因组DNA凝胶电泳

Fig. 5 Genomic DNA gel electrophoresis

表2 WESMS与马铃薯致病疫霉菌的Zeta电位

Table 2 Zeta potential of WESMS and Phytophthora infestans

样品

Sample

Zeta电位 Zeta potential/mV

最小值 Min

最大值 Max

平均值 Mean

菌糠提取液

WESMS

-30.76

-26.86

-29.25

致病疫霉菌

Phytophthora infestans

-38.86

-38.39

-38.57

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