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新型生防菌 111A-NL1 菌株所产挥发性化合物对草莓灰霉病的抑菌作用

Antifungal effects of volatile compounds produced by Tetrapisispora sp. strain 111A-NL1 as a new biocontrol agent on the strawberry grey mold disease.

机构信息

Department of Biology and Biotechnology, Faculty of Sciences, University of Kurdistan, Sanandaj, Iran.

Department of Plant Protection, Faculty of Agriculture, University of Kurdistan.

出版信息

Cell Mol Biol (Noisy-le-grand). 2022 Apr 30;68(4):12-23. doi: 10.14715/cmb/2022.68.4.2.

Abstract

An antagonistic yeast strain was isolated from the strawberry fruit cv. Paros, and its antifungal properties against Botrytis cinerea causal agent of strawberry grey mold disease were evaluated under in vitro and in vivo experiments. The isolate was tentatively identified as Tetrapisispora sp. strain 111A-NL1 based on phenotypic characteristics and sequence analysis of D1/D2 domains of the 26S rRNA gene. Volatile organic compounds (VOCs) produced by the 111A-NL1 strain inhibited the mycelial growth of fungal pathogen (75.19%) and conidial germination (63.34%); however, inhibition percentage of mycelial growth of pathogen by dual culture test was less (19.49%). Also, the strain produced pectinase, siderophore, chitinase, IAA, as well as gibberellin, and could solubilize phosphate. Additionally, the disease severity of strawberry grey mold was decreased by employing living cells and volatile metabolites methods by 47.61% and 74.05%, respectively, in comparison with untreated control seven days after inoculation. Therefore, its mode of action might consist of antibiosis and VOCs production by yeast strain 111A-NL1 against B. cinerea. The VOCs released by strain 111A-NL1 were analyzed, and thirty-three chemical compounds were determined by gas chromatography-mass spectroscopy (GC-MS). Out of them, Decane (12.79%), Squalene (9.60%), Undecane (7.98%), Benzene, 1,2,3-trimethyl- (7.67%), Nonane, 2,6-dimethyl- (5.69%), Benzene, 1-ethyl-3-methyl- (5.55%), Mesitylene (4.17%), and Phenylethyl Alcohol (3.33%) were the major components. In addition, the selected strain reduced natural decay incidence and weight loss of fruit, and preserved quality parameters of strawberry fruit including firmness, soluble solids content, and titratable acidity. This research averred, for the first time, that the creation of VOCs by Tetrapisispora sp. strain 111A-NL1 could play an essential role as a biofumigant in the antifungal activity against strawberry grey mold.

摘要

从草莓品种 Paros 中分离出一株拮抗酵母,并在体外和体内实验中评估其对草莓灰霉病病原菌 Botrytis cinerea 的抗真菌特性。根据表型特征和 26S rRNA 基因 D1/D2 区序列分析,该分离物被初步鉴定为 Tetrapisispora sp. 111A-NL1 菌株。111A-NL1 菌株产生的挥发性有机化合物(VOCs)抑制了真菌病原体的菌丝生长(75.19%)和分生孢子萌发(63.34%);然而,双层培养试验中病原体菌丝生长的抑制率较低(19.49%)。此外,该菌株还产生果胶酶、铁载体、几丁质酶、IAA 和赤霉素,并能溶解磷酸盐。此外,与未处理对照相比,接种后 7 天,采用活菌和挥发性代谢物方法,草莓灰霉病的严重程度分别降低了 47.61%和 74.05%。因此,酵母菌株 111A-NL1 对 B. cinerea 的作用模式可能包括抑菌作用和 VOCs 的产生。对 111A-NL1 菌株释放的 VOCs 进行了分析,并通过气相色谱-质谱联用(GC-MS)确定了 33 种化学物质。其中,癸烷(12.79%)、角鲨烯(9.60%)、十一烷(7.98%)、三甲苯(7.67%)、2,6-二甲基壬烷(5.69%)、1-乙基-3-甲基苯(5.55%)、均三甲苯(4.17%)和苯乙醇(3.33%)是主要成分。此外,该选定菌株降低了果实的自然腐烂发生率和失重率,并保持了草莓果实的品质参数,包括硬度、可溶性固形物含量和可滴定酸度。本研究首次证明, Tetrapisispora sp. 111A-NL1 产生的 VOCs 可能在其对草莓灰霉病的抗真菌活性中发挥重要作用,作为一种生物熏蒸剂。

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