异辛醇对灰葡萄孢的抑菌作用及转录组分析

doi:10.16409/j.cnki.2095-039x.2023.02.068

摘要/Abstract

摘要： 为探究异辛醇对灰葡萄孢Botrytis cinerea的抑制机理，采用双皿对扣法测定了异辛醇对灰葡萄孢的抑菌效果，利用扫描电镜技术观察了异辛醇处理后灰葡萄孢菌丝的形态变化，测定异辛醇对灰葡萄孢细胞膜胞外电导率和麦角甾醇合成的影响，并基于转录组测序技术（RNA sequencing，RNA-Seq）分析了异辛醇处理对灰葡萄孢的影响。结果表明：异辛醇对灰葡萄孢菌丝的抑菌率为67.92%。经异辛醇处理后，灰葡萄孢菌丝出现大量畸形、不规则变形扭曲，细胞膜通透性增加。异辛醇处理4、5和6 d后，细胞膜麦角甾醇的含量分别降低44.35%、38.82%和42.43%；蛋白质浓度分别降低65.70%、77.93%和58.57%。转录组测序共得到577个差异表达基因（DEGs），其中312个基因表达量上调，265个基因表达量下调。通过同源蛋白簇（COG）、基因本体论（GO）和京都基因与基因组百科全书（KEGG）功能分析，发现参与膜整体组分、麦角甾醇生物合成途径、氨基酸代谢和核糖体生物发生通路中的DEGs均显著下调，表明异辛醇可能通过影响灰葡萄孢的麦角甾醇含量、能量的产生和核糖体结构，从而起到对灰葡萄孢的抑菌作用。

关键词: 异辛醇, 灰葡萄孢, 抑菌作用, 转录组

Abstract: To reveal the inhibitory mechanism of isooctyl alcohol to Botrytis cinerea, the inhibition effect of isooctyl alcohol on B. cinerea was determined using two-sealed-base-plate method. Mycelial morphology treated with isooctyl alcohol was observed by scanning electron microscopy. The influence of isooctyl alcohol on the extracellular conductivity and the ergosterol synthesis were measured. Response of B. cinerea to isooctyl alcohol was analyzed by RNA sequencing (RNA-Seq). The results showed that the inhibition rate of isooctyl alcohol to mycelia was 67.92%. After treatment with isooctyl alcohol, lots of mycelia were deformed and distorted with the increasing of membrane permeability. At 4, 5 and 6 days after isooctyl alcohol treatment, the ergosterol contents in cell membrane decreased to 44.35%, 38.82% and 42.43%, and the protein concentration decreased to 65.70%, 77.93% and 58.57%, respectively. A total of 577 differentially expressed genes (DEGs), including 312 up-regulated genes and 265 down-regulated genes, were obtained by RNA-Seq. Based on functional analysis of clusters of orthologous groups (COG), gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG), DEGs involved in membrane integral components, ergosterol biosynthesis pathway, amino acid metabolism and ribosome biogenesis pathway were significantly down-regulated, indicating that isooctyl alcohol might affect ergosterol content, energy production and ribosome structure, and further play inhibit role in the growth of B. cinerea.

Key words: isooctyl alcohol, Botrytis cinerea, inhibition effect, RNA-Seq

中图分类号:

S436

段天坤, 王燕, 袁佳琪, 苏雅馨, 史璐欣, 王琳, 苏健, 王美琴, 王春伟. 异辛醇对灰葡萄孢的抑菌作用及转录组分析[J]. 中国生物防治学报, 2023, 39(6): 1434-1445.

DUAN Tiankun, WANG Yan, YUAN Jiaqi, SU Yaxin, SHI Luxin, WANG Lin, SU Jian, WANG Meiqin, WANG Chunwei. Inhibition Effect and Transcriptome Analysis of Isooctyl alcohol on Botrytis cinerea[J]. Chinese Journal of Biological Control, 2023, 39(6): 1434-1445.

参考文献

[1] Van Kan J A L, Stassen J H M, Mosbach A, et al. A gapless genome sequence of the fungus Botrytis cinerea[J]. Molecular Plant Pathology, 2017, 18(1): 75-89.
[2] 梁巧兰, 张娜, 魏列新, 等. 深绿木霉蛋白质TraT2A诱导兰州百合抗灰霉病的作用[J]. 中国生物防治学报, 2017, 33(4): 545-551.
[3] 魏新燕, 黄媛媛, 黄亚丽, 等. 拮抗灰霉菌的海洋细菌甲基营养型芽胞杆菌的筛选、鉴定及其抑菌活性物质的研究[J]. 中国生物防治学报, 2017, 33(5): 667-674.
[4] Cong M L, He S, Ma H J, et al. Hormetic effects of carbendazim on the virulence of Botrytis cinerea[J]. Plant Disease, 2018, 102(5): 886-891.
[5] 张正炜, 陈秀, 石小媛, 等. 我国农作物灰霉病杀菌剂的应用现状[J]. 中国蔬菜, 2021, 1(2): 41-46.
[6] 黄蓉, 胡建坤, 黄瑞荣. 酵母挥发性物质环辛四烯抑制灰霉菌的研究[J]. 中国生物防治学报, 2017, 33(2): 266-272.
[7] 王燕, 王春伟, 王琳, 等. 梨灰霉病菌拮抗细菌Pseudomonas extremorientalis的鉴定、发酵条件优化及防效评价[J]. 中国生物防治学报, 2019, 35(3): 437-448.
[8] 王凌宇, 廖晓兰, 张亚. 草莓灰霉病的防治研究进展[J]. 湖南农业科学, 2015, 357(6): 142-144.
[9] 肖景惠, 逄飞, 倪瑞琪, 等. 植物灰葡萄孢菌生物防治与化学防治机理的研究进展[J]. 中国蔬菜, 2019, 1(9): 18-23.
[10] Romanazzi G, Smilanick L J, Feliziani E, et al. Integrated management of postharvest gray mold on fruit crops[J]. Postharvest Biology and Technology, 2016, 113: 69-76.
[11] 陈宇春, 陈敏, 王其刚, 等. 植物灰霉病及抗性研究进展[J]. 江苏农业科学, 2020, 48(15): 42-51, 63.
[12] Zhou J Y, Li X, Zheng J Y, et al. Volatiles released by endophytic Pseudomonas fluorescens promoting the growth and volatile oil accumulation in Atractylodes lancea[J]. Plant Physiology and Biochemistry, 2016, 101: 132-140.
[13] 钟涛, 王智荣, 杜木英. 微生物源挥发性物质防治采后果蔬病害的研究进展[J]. 微生物学报, 2021, 61(7): 1771-1785.
[14] Liu W W, Zhao L J, Wang C, et al. Bioactive evaluation and application of antifungal volatiles generated by five soil bacteria[J]. Acta Phytophylacica Sinica, 2009, 36(2): 97-105.
[15] Huang R, Che H J, Zhang J, et al. Evaluation of Sporidiobolus pararoseus strain YCXT3 as biocontrol agent of Botrytis cinerea on post-harvest strawberry fruits[J]. Biological Control, 2012, 62(1): 53-63.
[16] Wang E Z, Liu X D, Si Z Y, et al. Volatile organic compounds from rice rhizosphere bacteria inhibit growth of the pathogen Rhizoctonia solani[J]. Agriculture, 2021, 11(4): 368.
[17] 董丽红, 郭庆港, 梁芸, 等. 枯草芽孢杆菌NCD-2挥发物对大丽轮枝菌的影响[C].//植物病理科技创新与绿色防控——中国植物病理学会, 2021年学术年会论文集, 2021: 626.
[18] 余璐, 魏琛, 张凯歌, 等. 异辛醇对稻谷中霉菌及其毒素的抑制作用研究[J]. 粮食与油脂, 2022, 35(5): 65-69, 79.
[19] 余璐, 魏琛, 张凯歌, 等. 枯草芽孢杆菌PW2产挥发性物质对赭曲霉的抑制作用[J]. 食品科学, 2023, 44(14): 125-133.
[20] 雷春妮, 张雅珩, 李经纬, 等. 不同品种与加工工艺对初榨橄榄油挥发性风味成分的影响[J]. 中国油脂, 2019, 44(10): 35-41, 54.
[21] 汪冬冬, 张其圣, 陈功, 等. 不同蔬菜原料发酵泡菜挥发性成分解析[J]. 食品工业科技, 2018, 39(3): 234-242.
[22] 中华人民共和国国家卫生和计划生育委员会. GB 2760-2014食品安全国家标准食品添加剂使用标准[S]. 北京: 中国标准出版社, 2015.
[23] Wang C W, Wang Y, Wang L, et al. Biocontrol potential of volatile organic compounds from Pseudomonas chlororaphis ZL3 against postharvest gray mold caused by Botrytis cinerea on Chinese cherry[J]. Biological Control, 2021, 159: 104613.
[24] 陈明月, 姜涛, 赵冬梅, 等. 先进的组学技术在植物抗病研究中的应用[J]. 中国农学通报, 2022, 38(24): 86-91.
[25] 吴诗媛. 基于组学技术的枯草芽孢杆菌CF-3 VOCs对炭疽菌和褐腐菌的影响及应用研究[D]. 上海: 上海大学, 2019.
[26] Wang C W, Duan T K, Shi L X, et al. Characterization of volatile organic compounds produced by Bacillus siamensis YJ15 and their antifungal activity against Botrytis cinerea[J]. Plant Disease, 2022, 106(9): 2321-2329.
[27] 曹林青, 詹发强, 高宇洁, 等. 嗜线虫致病杆菌抑制灰葡萄孢的效应[J]. 微生物学通报, 2021, 48(11): 4123-4133.
[28] Tian J, Wang Y Z, Zeng H, et al. Efficacy and possible mechanisms of perillaldehyde in control of Aspergillus niger causing grape decay[J]. International Journal of Food Microbiology, 2015, 202: 27-34.
[29] 刘亚苓, 于营, 鲁海坤, 等. 侧孢短芽孢杆菌S2-31拮抗下细辛叶枯病菌转录组差异表达分析[J]. 生物技术通报, 2021, 37(2): 111-121.
[30] 高雪, 劳广术, 谭峥, 等. 贝莱斯芽孢杆菌HN-2次生代谢产物处理下黄单胞菌（Xoo）的转录组分析[J]. 热带生物学报, 2023, 14(4): 389-398.
[31] 张瑶, 高弢, 马桂珍, 等. 基于转录组测序技术分析愈创木酚对禾谷镰刀菌的抑菌机制[J]. 江苏农业学报, 2022, 38(2): 343-351.
[32] 冯庭跃. 磺酰胺类化合物SYAUP-CN-26对灰葡萄孢菌作用机理研究[D]. 沈阳: 沈阳农业大学, 2020.
[33] Sant D G, Tupe S G, Ramana C V, et al. Fungal cell membrane-promising drug target for antifungal therapy[J]. Journal of Applied Microbiology, 2016, 121(6): 1498-1510.
[34] 张明. 白色念珠菌ARE2基因功能及两种天然产物的抗真菌机制研究[D]. 济南: 山东大学, 2018.
[35] 权雯婧, 刘超, 李奥, 等. 抗真菌剂三峡肽素抑制指状青霉的转录组学研究[J]. 食品工业科技, 2022, 43(6): 109-117.
[36] 严园园, 汪天明, 施高翔, 等. 黄连解毒汤联合氟康唑对耐药白念珠菌麦角甾醇的影响[J]. 中国中药杂志, 2015, 40(4): 727-732.
[37] 渠非. 木瓜海棠内生球毛壳菌MG2对苹果树腐烂病的生防作用研究[D]. 杨凌: 西北农林科技大学, 2022.
[38] Wang J, Chi Z Y, Zhao K, et al. A transcriptome analysis of the antibacterial mechanism of flavonoids from Sedum aizoon L. against Shewanella putrefaciens[J]. World Journal of Microbiology & Biotechnology, 2020, 36(7): 94.
[39] 李素平, 袁玲, 迟少艺, 等. 一种自溶性产酶溶杆菌新菌株LE16对温室番茄灰霉病的抑制作用[J]. 微生物学报, 2022, 62(10): 3871-3885.
[40] Ning Y, Ma M, Zhang Y, et al. Antibacterial mechanism of sucrose laurate against Bacillus cereus by attacking multiple targets and its application in milk beverage[J]. Food Research International, 2022, 154: 111018.
[41] 崔醒, 朱秋劲, 侯瑞, 等. 丁香酚、香芹酚和百里香酚对禾谷镰刀菌的抑菌活性及机制[J]. 食品科学, 2022, 43(23): 10-18.
[42] 何方涛. 茉莉酸甲酯诱导季也蒙毕赤酵母对苹果采后青霉病的控制及其机制研究[D]. 苏州: 江苏大学, 2020.
[43] Xu S Y, Yamamoto N. Anti-infective nitazoxanide disrupts transcription of ribosome biogenesis-related genes in yeast[J]. Genes & genomics, 2020, 42(8): 915-926.