纳他霉素对芒果采后胶孢炭疽菌的抑菌效果及机理

doi:10.11983/CBB19044

植物学报 ›› 2019, Vol. 54 ›› Issue (4): 455-463.DOI: 10.11983/CBB19044

所属专题：逆境生物学专辑 (2019年54卷2期)

纳他霉素对芒果采后胶孢炭疽菌的抑菌效果及机理

刘佳怡¹,王嘉欣¹,宋海超²,张正科¹,徐祥彬¹,吉训聪^3,^*(),史学群^1,^*()

1 海南大学食品学院, 海口 570228
2 海南大学热带农林学院, 海口 570228
3 海南省农业科学研究院植物保护研究所, 海口 571100

收稿日期:2019-03-08 接受日期:2019-05-06 出版日期:2019-07-10 发布日期:2020-01-08
通讯作者: 吉训聪,史学群
基金资助:
国家重点研发计划(2016YFD0400904)

Antifungal Activity and Mechanisms of Natamycin Against Colletotrichum gloeosporioides in Postharvest Mango Fruit

Jiayi Liu¹,Jiaxin Wang¹,Haichao Song²,Zhengke Zhang¹,Xiangbin Xu¹,Xuncong Ji^3,^*(),Xuequn Shi^1,^*()

1 College of Food Science and Technology, Hainan University, Haikou 570228, China
2 Institute of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China
3 Institute of Plant Protection, Hainan Academy of Agricultural Sciences, Haikou 571100, China

Received:2019-03-08 Accepted:2019-05-06 Online:2019-07-10 Published:2020-01-08
Contact: Xuncong Ji,Xuequn Shi

摘要/Abstract

摘要： 以纳他霉素为抑菌剂, 实验测定了离体条件下不同浓度纳他霉素对胶孢炭疽菌(Colletotrichum gloeosporioides)的孢子萌发及菌丝生长的抑制效果, 以及活体损伤接种炭疽病菌后, 纳他霉素对芒果(Mangifera indica)果实炭疽病的防治效果。通过测定纳他霉素处理后胶孢炭疽菌的细胞膜相对渗透率、可溶性蛋白含量、细胞膜完整性、孢子内活性氧水平和线粒体分布情况, 初步探明其抑菌机理。结果表明, 3 mg∙L ^-1纳他霉素可显著抑制胶孢炭疽菌孢子萌发、芽管伸长和菌落生长, 80 mg∙L ^-1纳他霉素可有效抑制芒果贮存过程中果实炭疽病斑的扩展。纳他霉素处理后胶孢炭疽菌细胞膜相对渗透率和可溶性蛋白含量增加; 2 mg∙L ^-1纳他霉素处理8小时, 处理组胶孢炭疽菌孢子细胞膜损伤染色率为33.6%, 对照组染色率为13.9%; 处理组胞内活性氧产生染色率达46.9%, 比对照组高39.7%; 同时观察到纳他霉素使胞内线粒体分布不均且荧光信号微弱。以上结果表明, 纳他霉素可以破坏胶孢炭疽病菌细胞膜, 诱导活性氧大量积累, 并降低线粒体活性, 从而干扰菌体正常生理活性, 使其代谢活动受影响, 从而达到抑菌目的。

关键词: 纳他霉素, 芒果, 胶孢炭疽病菌, 抑菌机理

Abstract: In this study, we examined the inhibitory effects of natamycin at different concentrations on the conidial germination and mycelial growth of Colletotrichum gloeosporioides in vitro as well as the controlled effect of natamycin on postharvest anthracnose of mango (Mangifera indica) fruit inoculated with C. gloeosporioides. To further explore the underlying antifungal mechanism, we analyzed the membrane permeability, soluble protein content, changes in cell membrane integrity, intracellular reactive oxygen species (ROS) level and mitochondrial distribution in C. gloeosporioides after natamycin treatment. Natamycin at 3 mg∙L ^-1 effectively suppressed the conidial germination, germ tube elongation and mycelial growth of C. gloeosporioides. Also, 80 mg∙L ^-1natamycin significantly inhibited the expansion of anthracnose lesions in mango fruit during storage. Furthermore, natamycin treatment increased the relative permeability and soluble protein content in the cell membrane of C. gloeosporioides. After 8h treatment with natamycin 2 mg∙L ^-1, the staining rate of damaged cell membranes in C. gloeosporioides was 33.6% and 13.9% in the control. The staining rate of intracellular ROS reached 46.9% in treated conidia, which was 39.7% higher than that of the control. Natamycin treatment caused heterogeneous distribution of intracellular mitochondria along with weaker fluorescence as compared with the control. In summary, natamycin can destroy the cell membrane of C. gloeosporioides, induce ROS accumulation and reduce mitochondrial activity, thus interfering in the normal physiological activity of C. gloeosporioides and affecting its metabolic activities.

Key words: natamycin, mango, Colletotrichum gloeosporioides, antifungal mechanism

刘佳怡,王嘉欣,宋海超,张正科,徐祥彬,吉训聪,史学群. 纳他霉素对芒果采后胶孢炭疽菌的抑菌效果及机理. 植物学报, 2019, 54(4): 455-463.

Jiayi Liu,Jiaxin Wang,Haichao Song,Zhengke Zhang,Xiangbin Xu,Xuncong Ji,Xuequn Shi. Antifungal Activity and Mechanisms of Natamycin Against Colletotrichum gloeosporioides in Postharvest Mango Fruit. Chinese Bulletin of Botany, 2019, 54(4): 455-463.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB19044

https://www.chinbullbotany.com/CN/Y2019/V54/I4/455

图/表 7

图1 纳他霉素对胶孢炭疽菌孢子萌发和芽管伸长的影响 (A) 孢子萌发显微图(Bars=50 μm); (B) 孢子萌发率; (C) 芽管长度。不同小写字母表示各处理组间差异显著(P<0.05)。

Figure 1 Effect of natamycin on conidial germination and germ tube elongation of Colletotrichum gloeosporioides (A) Micrograph of conidial germination (Bars=50 μm); (B) Conidial germination rate; (C) Germ tube length. Treatments followed by different lowercase letters are statistically different by the Duncan’s multiple range test (P<0.05).

图2 纳他霉素对胶孢炭疽菌菌丝生长的影响 (A) 菌落生长图(Bar=15 mm); (B) 菌落直径。不同小写字母表示各处理组间差异显著(P<0.05)。

Figure 2 Effect of natamycin on colony growth of Colletotrichum gloeosporioides (A) Colony morphology (Bar=15 mm); (B) Colony diameter. Treatments followed by different lowercase letters are statistically different by the Duncan’s multiple range test (P<0.05).

图3 纳他霉素对芒果采后炭疽病的影响 (A) 炭疽病发病情况(Bar=8 mm); (B) 炭疽病斑直径。不同小写字母表示各处理组间差异显著(P<0.05)。

Figure 3 Effect of natamycin on anthracnose in mango fruit (A) Symptoms of anthracnose in fruit (Bar=8 mm); (B) Lesion diameter of anthracnose. Treatments followed by different lowercase letters are statistically different by the Duncan’s multiple range test (P<0.05).

图4 纳他霉素对胶孢炭疽菌细胞膜透性和可溶性蛋白含量的影响 (A) 细胞膜相对渗透率; (B) 可溶性蛋白含量

Figure 4 Effect of natamycin on cell membrane permeability and soluble protein content of Colletotrichum gloeosporioides (A) Relative permeability of cell membrane; (B) Soluble protein content

图5 纳他霉素对胶孢炭疽菌孢子细胞膜完整性的影响 (A) 荧光显微镜下碘化丙啶(PI)染色情况, 细胞膜受损的孢子呈红色荧光(Bars=50 μm); (B) PI染色率。不同小写字母表示各处理组间差异显著(P<0.05)。

Figure 5 Effect of natamycin on membrane integrity of Colletotrichum gloeosporioides conidia (A) Propidium iodide (PI) staining under fluorescence microscope, conidia with damaged plasma membranes showed red fluorescence (Bars=50 μm); (B) PI staining rate. Treatments followed by different lowercase letters are statistically different by the Duncan’s multiple range test (P<0.05).

图6 纳他霉素对胶孢炭疽菌活性氧(ROS)含量的影响 (A) 荧光显微镜下DCFH-DA染色情况, 诱导活性氧产生的孢子呈绿色荧光(Bars=50 μm); (B) DCFH-DA染色率。不同小写字母表示各处理组间差异显著(P<0.05)。

Figure 6 Effect of natamycin on reactive oxygen species (ROS) content of Colletotrichum gloeosporioides (A) DCFH-DA staining under fluorescence microscope, conidia with intracellular ROS induction exhibit green fluorescence (Bars=50 μm); (B) DCFH-DA staining rate. Treatments followed by different lowercase letters are statistically different by the Duncan’s multiple range test (P<0.05).

图7 纳他霉素对胶孢炭疽菌孢子内线粒体分布的影响红色荧光处为线粒体聚集区(Bars=5 μm)。

Figure 7 Effect of natamycin on the distribution of mitochondria of Colletotrichum gloeosporioides conidia The red fluorescence represented the mitochondrial aggregation area (Bars=5 μm).

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纳他霉素对芒果采后胶孢炭疽菌的抑菌效果及机理

Antifungal Activity and Mechanisms of Natamycin Against Colletotrichum gloeosporioides in Postharvest Mango Fruit

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