抗真菌剂三峡肽素抑制指状青霉的转录组学研究  

Transcriptomics Study of Sanxiapeptin against Penicillium digitatum as an Antifungal

作　　者：权雯婧 刘超 李奥 薛艳红 刘士平 QUAN Wenjing;LIU Chao;LI Ao;XUE Yanhong;LIU Shiping(Key Laboratory of Functional Yeast(China National Light Industry),College of Life Science and Pharmacy,Three Gorges University,Yichang 443002,China)

机构地区：[1]三峡大学生物与制药学院,中国轻工业功能酵母重点实验室,湖北宜昌443002

出　　处：《食品工业科技》2022年第6期109-117,共9页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(31270389,31540065)。

摘　　要：为了分析三峡肽素(Sanxiapeptin)对柑橘类水果的主要采后致腐菌指状青霉(Penicillium digitatum)的抑制机理,本研究首先利用显微技术观察了指状青霉随三峡肽素的形态变化,并分析了细胞壁的保护剂山梨醇与细胞膜的保护剂麦角固醇在三峡肽素在抑制真菌过程中的作用,然后通过BGISEQ-500研究平台,对三峡肽素处理后的指状青霉进行了转录组测序,对测序产生的6.38 Gb序列数据进行了无参转录组分析。结果表明,经三峡肽素处理后,指状青霉的细胞表面形态发生了明显变化。与正对照膜靶点杀菌剂百可得不一样,细胞膜的保护剂麦角固醇可以明显缓解三峡肽素的抑制效果,而细胞壁保护剂山梨醇却会促进其抑菌能力。转录组序列相互比对率为93.66%,显示出较高的测序质量。指状青霉经三峡肽素处理后,超过2000个基因发生了表达变化,主要涉及细胞膜、细胞壁组分、自噬和糖代谢等途径。鉴于三峡肽素与百可得的作用途径不同,本研究选取了仅由三峡肽素引起的530个差异表达基因进行分析,发现这些基因主要与疏水蛋白、麦角固醇合成、聚糖合成及GPI(糖基磷脂酰肌醇,Glycosylphosphatidylinositol)的代谢相关,在17个候选基因中,有8个基因的RT-PCR半定量表达检测与转录组测序结果完全一致。上述结果说明三峡肽素可能通过影响细胞壁或细胞膜等细胞表面的功能,从而起到抑制柑橘采后致腐真菌的作用。To analyze the inhibitory mechanism of Sanxiapeptin on the main postharvest spoilage fungus P. digitatum of citrus fruits, the morphology responded to Sanxiapeptin was observed by microscopy. The cell wall protective agent sorbitol and cell membrane protective agent ergosterol were used to analyze the inhibitory effects of Sanxiapeptin on the fungi.Through BGISEQ-500 research platform, the transcriptome of P. digitatum treated by Sanxiapeptin was sequenced, and the6.38 Gb data were analyzed by non-parameter method. The results showed that the cell surface morphology changed significantly after treatment by Sanxiapeptin. Unlike the positive control Bellkute, a membrane-target antifungal, the cell membrane protector of ergosterol significantly alleviated the inhibitory effect of Sanxiapeptin, while the cell wall protector of sorbitol promoted the antifungal ability. The transcriptome alignment rate reached 93.66%, showing high sequencing quality. Sanxiapeptin affected the expression of more than 2000 genes in P. digitatum, which mainly involved in the components of cell membrane and cell wall, autophagy and sugar metabolism. In view of the different pathway from Bellkute, 530 differentially expressed genes caused by Sanxiapeptin alone were analyzed, and these genes were mainly related to the metabolism of hydrophobin, ergosterol, glycan and GPI(glycosylphosphatidylinositol). 17 candidate genes were selected for semi-quantitative expression verification by RT-PCR, among them, 8 genes were completely consistent with the transcriptome sequencing. All the results indicated that Sanxiapeptin could inhibit the postharvest decay fungi in citrus mediated by cell walls or cell membranes.

关 键 词：三峡肽素 指状青霉 抗真菌 转录组 柑橘 采后致腐 

分 类 号：Q939.92[生物学—微生物学]