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

doi:10.11983/CBB19044

Abstract

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

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[J]. Chinese Bulletin of Botany, 2019, 54(4): 455-463.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB19044

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

Figures/Tables 7

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).

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).

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).

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

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).

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).

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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