Article(id=1276616481303564897, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276616263778562546, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.11.016, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1751040000000, receivedDateStr=2025-06-28, revisedDate=null, revisedDateStr=null, acceptedDate=1754323200000, acceptedDateStr=2025-08-05, onlineDate=1782298687529, onlineDateStr=2026-06-24, pubDate=1764000000000, pubDateStr=2025-11-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782298687529, onlineIssueDateStr=2026-06-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782298687529, creator=13701087609, updateTime=1782298687529, updator=13701087609, issue=Issue{id=1276616263778562546, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='11', pageStart='2549', pageEnd='2815', issueExtLink='null', onlineDate='null', pubDate='1764000000000', pubDateStr='2025-11-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782298635668, creator='13701087609', updateTime=1782299117657, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276618285483426694, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276616263778562546, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276618285487620999, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276616263778562546, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2713, endPage=2721, ext={EN=ArticleExt(id=1276616482834485859, articleId=1276616481303564897, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Effects of Phospholipase Inhibitors on the Growth of Colletotrichum musae and Phospholipase Activity, columnId=1236292524264968282, journalTitle=Chinese Journal of Tropical Crops, columnName=Plant Protection & Bio-safety, runingTitle=null, highlight=null, articleAbstract=

Banana anthracnose, caused by Colletotrichum musae, is a major postharvest disease that threatens fruit quality and banana industry sustainability. Phospholipase (PL) plays dual roles in plant disease resistance and pathogen metabolism, acting as a key enzyme in defense responses and as a virulence factor for pathogenic fungi.. Previous studies have shown that PL inhibitors can help maintaining membrane stability under stress conditions. In this study, the inhibitory effects of three PL inhibitors, i.e. quinacrine dihydrochloride (QU), neomycin sulfate (NE), and n-hexanol (Hex), on C. musae were evaluated through in vitro antifungal assays. Results showed that all three inhibitors significantly suppressed mycelial growth and spore production of C. musae. The half-maximal inhibitory concentrations (IC50) for mycelial growth were 1.00 mmol/L (QU), 5.00 mmol/L (NE), and 8.80 mmol/L (Hex). Enzymatic activity assays revealed that QU significantly inhibited both PLA activity in C. musae and banana peel tissues, NE primarily suppressed PLC activity in banana peel, whereas Hex markedly reduced PLD activity in C. musae. Moreover, the appropriate concentrations of the inhibitors (2.00 mmol/L QU, 3.00 mmol/L NE, and 13.20 mmol/L Hex) effectively controlled the development of banana anthracnose. The findings suggested that QU, NE, and Hex suppressed banana anthracnose through distinct mechanisms involving interference with host-pathogen interactions. This study would provide a theoretical basis for the green control of postharvest banana diseases via the targeted regulation of phospholipase activity.

, authors=null, authorsList=Hao DONG, Chunxia HUANG, Qiuyu CHENG, Lihong XIE, Ping YI, Shupei WANG, Qi WEI, Fang HUANG, Yuhan LONG, Li LI, Jian SUN, authorCompany=null, correspAuthors=Li LI, Jian SUN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1276616484096971377, articleId=1276616481303564897, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=磷脂酶抑制剂对香蕉炭疽菌生长及磷脂酶活性的影响, columnId=1236292524520820846, journalTitle=热带作物学报, columnName=植物保护与生物安全, runingTitle=null, highlight=null, articleAbstract=

由香蕉炭疽菌(Colletotrichum musae)引发的炭疽病是采后香蕉贮运期间的重要病害,影响果实品质与产业可持续发展。磷脂酶(PL)在植物抗病反应和病原菌代谢中具有双重作用,既是植物防御反应的关键酶,也是病原真菌的毒力因子,在逆境胁迫下使用PL抑制剂有助于维持细胞膜稳定性。本研究通过体外抑菌实验测定3种PL抑制剂——喹吖因二盐酸盐(QU)、新霉素硫酸盐(NE)和正己醇(Hex)对香蕉炭疽菌的抑制效果。结果显示:三者对香蕉炭疽菌菌丝生长和孢子生成均有显著抑制效果,半抑制浓度(IC50)分别为1.00、5.00、8.80 mmol/L。PL酶活性抑制试验显示,QU显著抑制香蕉炭疽菌及香蕉果皮PLA活性,NE显著抑制果皮PLC活性,Hex则显著抑制香蕉炭疽菌的PLD活性。此外,适宜浓度的3种PL抑制剂(2.00 mmol/L QU、3.00 mmol/L NE、13.20 mmol/L Hex)均能防控香蕉炭疽病的发展。研究结果表明,QU、NE和Hex通过不同机制干扰病原菌与寄主互作,从而有效控制香蕉炭疽病病害,为香蕉采后病害绿色防控提供理论依据。

, authors=

董昊(2000—),男,硕士研究生,研究方向:农产品贮藏与加工。

, authorsList=董昊, 黄春霞, 程秋芋, 谢丽红, 易萍, 王淑培, 韦琦, 黄方, 隆宇涵, 李丽, 孙健, authorCompany=null, correspAuthors=李丽, 孙健, authorNote=null, correspAuthorsNote=
* 李丽(1983—),E-mail:
孙健(1978—),E-mail:
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董昊(2000—),男,硕士研究生,研究方向:农产品贮藏与加工。

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董昊(2000—),男,硕士研究生,研究方向:农产品贮藏与加工。

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不同小写字母表示处理间差异显著(P<0.05)。

, figureFileSmall=7RGbSr96dcFFsosHU7pyQA==, figureFileBig=DcXDwEyFdcG+EWgC26UxIg==, tableContent=null), ArticleFig(id=1276616496365310660, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=EN, label=Fig. 2, caption=Effects of different concentrations of QU, NE and Hex on sporulation of C. musae, figureFileSmall=efmLDRuJHl1Aoz2IpG5T6Q==, figureFileBig=JG65eLPASEh60uZU4DASNQ==, tableContent=null), ArticleFig(id=1276616496432419525, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=CN, label=图2, caption=不同浓度的QU、NE和Hex对香蕉炭疽菌产孢量的影响

不同小写字母表示处理间差异显著(P<0.05)。

, figureFileSmall=efmLDRuJHl1Aoz2IpG5T6Q==, figureFileBig=JG65eLPASEh60uZU4DASNQ==, tableContent=null), ArticleFig(id=1276616496499528390, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=EN, label=Fig. 3, caption=Determination of different concentrations of QU, NE, and Hex on mycelial growth inhibition rate and half-maximal inhibitory concentration (IC50) of C. musae, figureFileSmall=au4YErgLp2zfMpwhyUv9wQ==, figureFileBig=4xGK2x7K8GzB88OKeFKM8Q==, tableContent=null), ArticleFig(id=1276616496570831559, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=CN, label=图3, caption=QU、NE和Hex对香蕉炭疽菌菌丝生长抑制率及半数抑制浓度(IC50)的影响, figureFileSmall=au4YErgLp2zfMpwhyUv9wQ==, figureFileBig=4xGK2x7K8GzB88OKeFKM8Q==, tableContent=null), ArticleFig(id=1276616496633746120, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=EN, label=Fig. 4, caption=Effects of different concentrations of QU, NE and Hex on lesion diameter of banana anthracnose, figureFileSmall=cqF6zJJNJCI97w9u1Ut/Rw==, figureFileBig=//nlof2SZ7FTkknCvijeMQ==, tableContent=null), ArticleFig(id=1276616496709243593, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=CN, label=图4, caption=不同浓度QU、NE和Hex处理对香蕉炭疽病病斑直径的影响

不同小写字母表示处理间差异显著(P<0.05)。

, figureFileSmall=cqF6zJJNJCI97w9u1Ut/Rw==, figureFileBig=//nlof2SZ7FTkknCvijeMQ==, tableContent=null), ArticleFig(id=1276616496772158154, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=EN, label=Fig. 5, caption=Effect of inhibitors QU, NE, and Hex on phospholipase activity of C. musae and banana fruit, figureFileSmall=y1DtclNKou3bZNYbC0AlkQ==, figureFileBig=W9hfBhDXAD+4NcweXtOAQw==, tableContent=null), ArticleFig(id=1276616496835072715, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276616481303564897, language=CN, label=图5, caption=QU、NE、Hex对香蕉炭疽菌和香蕉果实PL活性的影响

不同小写字母表示处理间差异显著(P<0.05)。

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磷脂酶抑制剂对香蕉炭疽菌生长及磷脂酶活性的影响
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董昊 1 , 黄春霞 2, 3 , 程秋芋 2 , 谢丽红 2 , 易萍 2, 3 , 王淑培 2, 3 , 韦琦 1 , 黄方 1 , 隆宇涵 2 , 李丽 1, 2, * , 孙健 2, 3, *
热带作物学报 | 植物保护与生物安全 2025,46(11): 2713-2721
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热带作物学报 |植物保护与生物安全 2025 , 46 (11) : 2713 -2721
磷脂酶抑制剂对香蕉炭疽菌生长及磷脂酶活性的影响
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董昊1, 黄春霞2, 3, 程秋芋2, 谢丽红2, 易萍2, 3, 王淑培2, 3, 韦琦1, 黄方1, 隆宇涵2, 李丽1, 2, * , 孙健2, 3, *
作者信息
  • 1.广西大学,广西南宁 530004
  • 2.广西壮族自治区农业科学院,广西南宁 530007
  • 3.广西果蔬贮藏与加工新技术重点实验室,广西南宁 530007
通讯作者:
* 李丽(1983—),E-mail:
孙健(1978—),E-mail:
Effects of Phospholipase Inhibitors on the Growth of Colletotrichum musae and Phospholipase Activity
Hao DONG1, Chunxia HUANG2, 3, Qiuyu CHENG2, Lihong XIE2, Ping YI2, 3, Shupei WANG2, 3, Qi WEI1, Fang HUANG1, Yuhan LONG2, Li LI1, 2, * , Jian SUN2, 3, *
Affiliations
  • 1.Guangxi University, Nanning, Guangxi 530004, China
  • 2.Guangxi Academy of Agricultural Sciences, Nanning, Guangxi 530007, China
  • 3.Guangxi Key Laboratory of Fruit and Vegetable Storage-processing Technology, Nanning, Guangxi 530007, China
出版时间: 2025-11-25 doi: 10.3969/j.issn.1000-2561.2025.11.016
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由香蕉炭疽菌(Colletotrichum musae)引发的炭疽病是采后香蕉贮运期间的重要病害,影响果实品质与产业可持续发展。磷脂酶(PL)在植物抗病反应和病原菌代谢中具有双重作用,既是植物防御反应的关键酶,也是病原真菌的毒力因子,在逆境胁迫下使用PL抑制剂有助于维持细胞膜稳定性。本研究通过体外抑菌实验测定3种PL抑制剂——喹吖因二盐酸盐(QU)、新霉素硫酸盐(NE)和正己醇(Hex)对香蕉炭疽菌的抑制效果。结果显示:三者对香蕉炭疽菌菌丝生长和孢子生成均有显著抑制效果,半抑制浓度(IC50)分别为1.00、5.00、8.80 mmol/L。PL酶活性抑制试验显示,QU显著抑制香蕉炭疽菌及香蕉果皮PLA活性,NE显著抑制果皮PLC活性,Hex则显著抑制香蕉炭疽菌的PLD活性。此外,适宜浓度的3种PL抑制剂(2.00 mmol/L QU、3.00 mmol/L NE、13.20 mmol/L Hex)均能防控香蕉炭疽病的发展。研究结果表明,QU、NE和Hex通过不同机制干扰病原菌与寄主互作,从而有效控制香蕉炭疽病病害,为香蕉采后病害绿色防控提供理论依据。

香蕉  /  炭疽病菌  /  磷脂酶抑制剂  /  磷脂酶活性

Banana anthracnose, caused by Colletotrichum musae, is a major postharvest disease that threatens fruit quality and banana industry sustainability. Phospholipase (PL) plays dual roles in plant disease resistance and pathogen metabolism, acting as a key enzyme in defense responses and as a virulence factor for pathogenic fungi.. Previous studies have shown that PL inhibitors can help maintaining membrane stability under stress conditions. In this study, the inhibitory effects of three PL inhibitors, i.e. quinacrine dihydrochloride (QU), neomycin sulfate (NE), and n-hexanol (Hex), on C. musae were evaluated through in vitro antifungal assays. Results showed that all three inhibitors significantly suppressed mycelial growth and spore production of C. musae. The half-maximal inhibitory concentrations (IC50) for mycelial growth were 1.00 mmol/L (QU), 5.00 mmol/L (NE), and 8.80 mmol/L (Hex). Enzymatic activity assays revealed that QU significantly inhibited both PLA activity in C. musae and banana peel tissues, NE primarily suppressed PLC activity in banana peel, whereas Hex markedly reduced PLD activity in C. musae. Moreover, the appropriate concentrations of the inhibitors (2.00 mmol/L QU, 3.00 mmol/L NE, and 13.20 mmol/L Hex) effectively controlled the development of banana anthracnose. The findings suggested that QU, NE, and Hex suppressed banana anthracnose through distinct mechanisms involving interference with host-pathogen interactions. This study would provide a theoretical basis for the green control of postharvest banana diseases via the targeted regulation of phospholipase activity.

banana  /  Colletotrichum musae  /  phospholipase inhibitors  /  phospholipase activity
董昊, 黄春霞, 程秋芋, 谢丽红, 易萍, 王淑培, 韦琦, 黄方, 隆宇涵, 李丽, 孙健. 磷脂酶抑制剂对香蕉炭疽菌生长及磷脂酶活性的影响. 热带作物学报, 2025 , 46 (11) : 2713 -2721 . DOI: 10.3969/j.issn.1000-2561.2025.11.016
Hao DONG, Chunxia HUANG, Qiuyu CHENG, Lihong XIE, Ping YI, Shupei WANG, Qi WEI, Fang HUANG, Yuhan LONG, Li LI, Jian SUN. Effects of Phospholipase Inhibitors on the Growth of Colletotrichum musae and Phospholipase Activity[J]. Chinese Journal of Tropical Crops, 2025 , 46 (11) : 2713 -2721 . DOI: 10.3969/j.issn.1000-2561.2025.11.016
香蕉(Musa acuminate L.)是芭蕉科芭蕉属多年生草本植物的果实,广泛分布于热带和亚热带地区,是全球最重要的水果和经济作物之一[1]。香蕉炭疽病是由香蕉炭疽菌(Colletotrichum musae)引起的一种主要病害,炭疽菌在香蕉青果期通过附着胞完成初侵染,随后以潜伏状态长期隐匿于果皮组织中,直至果实进入黄熟阶段才显症爆发,且成熟度与病情呈正相关,其潜伏特性导致传统采后防腐措施难以奏效,增加了贮运期病害防控难度[1-3]。在香蕉应对病原侵染的免疫响应中,细胞膜作为原生质界面的初级防御屏障,通过膜系统信号感知机制快速启动抗病应答。其中磷脂酶(PL)通过特异性水解膜磷脂(如磷脂酰肌醇)的甘油酯键,生成次级信号分子(如肌醇磷酸、二酰甘油等),激活钙信号级联和蛋白激酶网络,进而调控抗病相关基因表达及活性氧爆发等系统性防御反应[4-5]。PL介导的防御机制在保护植物的同时,可能引发一系列细胞损伤,其水解作用不仅导致细胞膜磷脂的降解,更会通过激活脂质过氧化反应链,引发氧化应激效应。这种级联反应会进一步加剧膜脂代谢紊乱,破坏膜系统稳态,最终造成细胞膜结构损伤和功能性障碍[6]
在采后逆境胁迫条件下,外源PL抑制剂可通过抑制膜脂降解相关酶活性,有效维持细胞膜结构的稳定,进而延缓果蔬采后品质劣变并延长其贮藏保鲜期[7-9]。喹吖因二盐酸盐(quinacrine dihydrochloride,QU),作为非特异性磷脂酶A2(PLA2)抑制剂,通过与PLA2活性位点结合,抑制其酶活性,减少磷脂水解,保护细胞膜完整性[10-11]。此外,QU还具有抑制烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶活性的作用,这对于减轻氧化应激、保护细胞免受损伤具有重要意义。新霉素硫酸盐(neomycin sulfate,NE)是一种磷脂酶C(PLC)的抑制剂,可与膜上的磷脂酰肌醇4,5-二磷酸(PIP2)竞争性结合,通过静电屏蔽作用,阻止PLC对PIP2的水解,进而抑制PLC的活性[12-14]。正己醇(hexanol,Hex)是一种饱和脂肪醇,具有显著的疏水亲脂特性,广泛应用于植物和动物的生理生化研究中,尤其在磷脂酶D(PLD)信号通路的研究中备受关注[10]。研究表明,PLD在催化磷脂生成磷脂酸的过程中,醇类化合物能够与磷脂的头部基团竞争性结合,进而抑制PLD的活性[15-16]
近年来,PL抑制剂在植物抗病研究中备受关注。但目前关于PLA、PLC和PLD抑制剂在采后果实上的作用效果尚缺乏系统全面的研究。本研究以香蕉炭疽菌和香蕉果实为材料,选用喹吖因二盐酸盐(QU)、新霉素硫酸盐(NE)和正己醇(Hex)作为PL的抑制剂,分析不同浓度磷脂酶抑制剂对菌丝生长和产孢量的影响,明确半抑制浓度(IC50);同时验证PL抑制剂对香蕉果实以及香蕉炭疽菌的PL活性的影响,进一步探究香蕉炭疽菌胁迫下不同浓度PL抑制剂对香蕉果实炭疽病的防控效果,以期为研究香蕉采后炭疽病的绿色防治提供理论依据。
(1)病原菌:香蕉炭疽菌(Colletotrichum musae 31249)为广西果蔬贮藏与加工新技术重点实验室保存菌株。将香蕉炭疽菌接种于马铃薯葡萄糖琼脂培养基(PDA)中28 ℃培养5 d进行活化。
(2)果实:实验选用的香蕉品种为桂蕉1号(Musa acuminata AAA cv. Guijiao No.1)。选取广西南宁市坛洛镇果园中7~8成熟的香蕉果实进行采收,采收后立即运至广西果蔬贮藏与加工新技术重点实验室。挑选大小一致、成熟度均匀、无机械损伤和病虫害的香蕉果实,清洗干净表面污物后用0.5%次氯酸钠溶液浸泡30 s进行表面消毒,再次用自来水冲洗干净后室温晾干备用。
(3)主要试剂:喹吖因二盐酸(QU)及新霉素硫酸盐(NE),生物技术级;正己醇(Hex),分析纯,上海麦克林生化科技股份有限公司。磷脂酶PLA、PLC及PLD酶联免疫分析试剂盒,上海恒远生物科技有限公司。
(4)主要仪器设备:SQ510C高压灭菌锅,重庆雅马拓科技有限公司;J-301高速冷冻离心机,德国Beckman公司;Synergy H1多功能酶标仪,美国Bio Tek公司;OSE-Y30/50组织研磨器,北京天根生化科技有限公司;CX31光学显微镜,日本OLYMPUS公司。
(1)将配置好的QU、NE和Hex母液与PDA培养基混匀,使QU的终浓度为0.01、0.10、0.50、1.00、1.50、2.00 mmol/L;NE的终浓度为0.50、1.00、1.50、2.00、2.50、3.00 mmol/L;Hex的终浓度为1.10、2.20、4.40、8.80、11.00、13.20 mmol/L。以不含任何PL抑制剂的PDA为对照组培养基(CK),将直径为5 mm的香蕉炭疽菌菌饼接入各处理组PDA培养基的中央,置于25 ℃恒温培养箱中培养,观察并测定第5天的菌落直径。
(2)将配置好的QU、NE和Hex母液与马铃薯葡萄糖肉汤(PDB)培养基混匀,使QU终浓度为0.01、0.10、0.50、1.00、1.50、2.00 mmol/L;NE终浓度为0.50、1.00、1.50、2.00、2.50、3.00 mmol/L;Hex终浓度为1.10、2.20、4.40、8.80、11.00、13.20 mmol/L。以不含任何PL抑制剂的PDB为对照组,将孢子悬浮液和PDB培养基以体积比(V/V)为1∶1000混匀后置于28 ℃、200 r/min摇床振荡培养,光学显微镜下测定第5天的孢子数量。
(3)计算QU、NE和Hex对香蕉炭疽菌菌丝生长的抑制率,以PL抑制剂浓度为横坐标、抑菌率为纵坐标,绘制量效关系曲线;并计算QU、NE和Hex对香蕉炭疽菌的抑制效应的IC50值,计算公式[17]为:
(1)将灭菌后的PDA培养基倒入培养皿,待凝固后在培养基上铺无菌玻璃纸,再在中央接入香蕉炭疽菌菌饼[18],28 ℃恒温培养5 d后,收集菌丝备用。随后迅速将菌丝在液氮中冻透,用高温灭菌的研钵磨碎。称取0.01 g菌丝放入1.50 mL无酶离心管,加100 μL pH 7.8的PBS缓冲液,研磨,在4 ℃、4500 r/min条件下离心15 min,取上清液(含PL的粗酶液)备用。分别取3份20 μL粗酶液置于1.5 mL离心管中,分别加入QU、NE和Hex母液,使抑制剂终浓度为PL抑制剂对香蕉炭疽菌作用下的IC50,室温静置5 min。根据试剂盒的操作说明,采用ELISA法测定香蕉炭疽菌的PLA、PLC和PLD活性。
(2)从未经处理的香蕉果皮鲜样上,用刀片切取2 cm×2 cm的果皮样本,用液氮快速冷冻后,置于-80 ℃保存备用。称取0.10 g研磨成粉的香蕉皮样品,加入1.0 mL pH 7.8的PBS缓冲液,立即混匀。随后在4 ℃、4500 r/min条件下离心15 min,收集上清液(含PL的粗酶液)备用。分别取3份20 μL粗酶液置于1.5 mL离心管中,分别向每份粗酶液中加入QU、NE和Hex母液,使体系中抑制剂的终浓度达到PL抑制剂对香蕉炭疽菌作用下的5倍IC50,室温静置5 min。最后,采用ELISA法测定香蕉果皮中的PLA、PLC和PLD活性。
将香蕉果实随机分为15组,分别为:QU处理组(0.00、1.00、2.00、3.00、4.00 mmol/L),NE处理组(0.00、2.00、3.00、4.00、5.00 mmol/L),Hex处理组(0.00、11.00、13.20、26.40、52.80 mmol/L)。每组12个香蕉果实。
对香蕉果实进行针刺接菌处理:使用无菌打孔器在香蕉中部开始,每隔3 cm打3个孔,每个孔深2 mm,直径2 mm。每个伤口接入20 μL经0.22 µm Millipore过滤器过滤的PL抑制剂溶液。待PL抑制剂完全被香蕉果实吸收后,再接入10 μL香蕉炭疽菌孢子悬液(1×106 spores/mL)。待香蕉果实完全吸收孢子悬液后,用厚度为0.03 mm的聚乙烯薄膜袋密封,置于28 ℃培养箱中贮藏,相对湿度保持在85%~90%之间。在第0、1、3、5天,分别拍照观察并测量病斑直径。
每个试验重复3次。使用Excel 2019软件进行数据分析处理,使用Origin 2022(Origin Lab,Northampton,MA,USA)和Grapad Prism 9.5.1(GraphPad,San Diego,California,USA)软件制图。使用SPSS 26(IBM Corp.,Armonk,New York,USA)软件进行方差分析,并用Duncan’s法检验组间差异显著性(P<0.05)。
为探究PL抑制剂对香蕉炭疽菌生长的抑制效果,本研究测定了不同浓度PL抑制剂处理后香蕉炭疽菌在PDA上的生长情况。如图1所示,培养5 d后,随着QU、NE、Hex浓度的升高,香蕉炭疽菌菌落逐渐变小,当QU、NE、Hex的浓度分别为2.00、3.00、13.20 mmol/L时,菌落生长受到最大程度的抑制,与对照组相比,菌落直径分别减小72.17%、38.26%、74.96%。
通过对添加了PL抑制剂的PDB培养基中香蕉炭疽菌孢子生成量的检测,发现施加抑制剂后的香蕉炭疽菌的孢子生成量显著减少,不同浓度的QU、NE、Hex均可显著抑制孢子产量,其中0.50 mmol/L QU、0.50 mmol/L NE、13.20 mmol/L Hex分别使孢子产量下降99.51%、88.76%、97.94%,说明达到一定浓度时,QU、NE、Hex均可抑制香蕉炭疽菌的产孢过程(图2)。
通过计算并统计不同浓度下QU、NE、Hex对香蕉炭疽菌菌落抑制率的变化,绘制抑菌率与浓度的计量关系图(图3)。QU的IC50为1.00 mmol/L,NE的IC50为5.00 mmol/L,Hex的IC50为8.80 mmol/L。
PL在植物生长发育中,具有防御作用,但其过表达会导致细胞膜磷脂降解,破坏膜的完整性,使病原菌更容易入侵寄主,从而加剧病害。为了探究PL抑制剂在病害防控中的潜在应用,本研究分析了不同浓度PL抑制剂处理的香蕉果实在香蕉炭疽菌胁迫下的病害发展情况,筛选出能够显著减缓病害的最佳抑制剂浓度。病斑直径作为发病严重程度的指标,通过观测接种香蕉炭疽菌并贮藏一定时间后香蕉果实病斑直径的大小,可以直接反映不同PL抑制剂对香蕉炭疽病的防控效果。从图4可知,不同PL抑制剂在一定的浓度下均可以有效防控香蕉炭疽病的发展,其中2.00 mmol/L QU、3.00 mmol/L NE、13.20 mmol/L Hex对香蕉炭疽病的防控效果最佳。随着抑制剂浓度持续升高,3种抑制剂对香蕉炭疽病的抑制效果均下降,尤其当NE浓度增至4.00 mmol/L时,果实病斑直径反而增大。
炭疽菌磷脂酶活性体外抑制试验结果(图5)显示,QU和Hex分别抑制香蕉炭疽菌的PLA和PLD的活性,且相较于对照组,1.00 mmol/L QU处理的菌丝样品PLA活性下降了18.41%,8.80 mmol/L Hex处理的菌丝样品PLD活性下降了18.66%,而5.00 mmol/LNE对香蕉炭疽菌的PLC活性无显著抑制作用。香蕉果皮磷脂酶活性的体外抑制结果(图5)显示,QU和NE分别抑制贮藏0、3 d的香蕉果皮的PLA和PLC活性,而Hex对香蕉果皮的PLD无抑制作用,反而显著提高了PLD的活性。
磷脂酶在植物生长发育及逆境胁迫中扮演关键角色,其表达受植物激素信号转导调控。近年来,关于植物逆境胁迫下磷脂酶表达与活性的研究已取得初步进展[19]。ZHANG等[20]研究发现,棉花PLA1-32基因上调表达以应对盐胁迫逆境。LIU等[21]研究发现,在低温和盐胁迫下番茄植株PLC4PLC3表达水平显著升高,揭示该类基因在应答多种非生物胁迫中具有潜在功能。SHUAI等[22]研究发现,香蕉在接种炭疽病菌后,PLC-1的表达水平显著提高,PLC的活性增强,同时伴随二酰甘油(DAG)和磷脂酸(PA)含量上升。上述研究结果共同说明植物通过上调PL表达水平来响应逆境胁迫。FANG等[6]系统综述了参与植物生物胁迫抗性和真菌致病性的PLCs的结构、分类及其生物学功能。然而,植物PL基因的过量表达可能会导致磷脂酶活性异常升高,从而加速细胞膜磷脂的降解,破坏细胞膜的完整性,进而为病原菌入侵提供有利条件[23-24]。彭慧等[25]发现,扩展青霉Penicillium expansum侵染苹果后,苹果果实的磷脂酶活性显著升高,膜磷脂降解速率加快,细胞膜的通透性增加,完整度降低。磷脂酶的作用是双重的,既能作为植物应对逆境胁迫的关键酶,也会因过度激活而破坏细胞膜完整性。
已有研究证实外源PL抑制剂能有效抑制磷脂酶的活性及表达水平,维持果实细胞膜结构的完整性,从而延长贮藏期;同时降低病原菌的致病性[26]。但关于PL抑制剂在果实抗病机制中的研究仍处于初步阶段。目前尚不清楚在病原菌侵染初期,PLA抑制剂是通过直接抑制病原菌致病性,还是通过间接调节果实抗性信号通路来影响病害的发生与发展。本研究探究了QU、NE和Hex三种PL抑制剂对香蕉炭疽菌生长及果实炭疽病的防治效果,并计算半数抑菌浓度IC50,评估三者作为PL抑制剂的潜在应用价值。QU、NE和Hex均显著抑制香蕉炭疽菌菌丝生长和孢子生成,对应的IC50值分别为1.00、5.00、8.80 mmol/L,表明PL抑制剂可以通过干扰病原菌的生长代谢过程从而实现对果实病害的有效防控。与本研究结果类似,ZHANG等[26]将PLD特异性抑制剂正丁醇施加到粉红单端孢Trichothecium roseum侵染的甜瓜果实上,发现正丁醇不仅显著降低了甜瓜果实PLD的活性,同时还下调了T. roseum中PLD相关基因的表达水平。这一结果证实PL抑制剂在果实病害防控中的潜在作用机制,即通过调节病原菌和果实细胞内的磷脂酶活性及相关基因表达,从而影响病害的发展进程。
在PL活性方面,QU显著抑制了香蕉炭疽菌和香蕉果皮的PLA活性;NE对香蕉炭疽菌的PLC活性无显著抑制效应,但能显著抑制香蕉果皮PLC的活性;而Hex则显著抑制香蕉炭疽菌的PLD活性,对香蕉果皮PLD活性则无明显影响。由此可见,不同类型抑制剂通过选择性地抑制不同PL的活性进而提高香蕉的抗病害能力。在植物受到病原菌侵染,PL表达水平通常会上调以响应病原菌反应。然而过高的PL表达会加速细胞膜降解,破坏细胞膜的完整性,进而加剧果实组织的损伤。本研究发现QU和NE可通过抑制香蕉果皮PL的活性,有效维持细胞膜完整性,从而提高香蕉的抗炭疽病的能力。而对于香蕉炭疽菌中PL来说,已有研究表明其在真菌信号传导及生长发育过程中发挥重要作用[27],为进一步探讨外源PL抑制剂对香蕉炭疽病的防控效果,本研究采用不同浓度的3种抑制剂QU、NE和Hex处理香蕉果实,通过损伤接种香蕉炭疽菌试验发现,适宜浓度的PL抑制剂均能在一定程度上抑制炭疽病害的病斑扩展,其中最佳处理浓度分别为:2.00 mmol/L QU、3.00 mmol/L NE、13.20 mmol/L Hex。随着抑制剂浓度的进一步升高,香蕉果实炭疽病病斑直径显著增大。对于QU和NE而言,过高的浓度可能通过严重抑制果实内相应磷脂酶的活性,干扰其介导的生物胁迫响应信号通路(如病原菌侵染),最终削弱果实的防御能力;与之相反,高浓度Hex可能因其对PLD活性的异常促进作用,破坏细胞膜完整性,继而加剧果实组织的病理损伤。
综合本研究结果,PL抑制剂QU、NE和Hex均能抑制香蕉炭疽菌的菌丝生长、孢子产量和调控PL活性,推测QU可能通过干扰香蕉炭疽菌自身的PL代谢并调节果皮PL活性来增强香蕉抗病性,抑制香蕉炭疽病的发生;NE则可能依赖增强植物抗性,通过抑制PLC活性以减少磷脂降解,进而提抗病性;Hex可能直接作用于病原菌,通过破坏香蕉炭疽菌膜代谢,从而抑制病原菌的侵染与扩展。但是,3种PL抑制剂在抑制香蕉炭疽病方面可能具有不同作用模式,其与病原菌和寄主磷脂酶的互作机制仍有待进一步探究。
  • 广西科技基地和人才专项(桂科AD24010045)
  • 国家自然科学基金项目(32272784; 32160732)
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2025年第46卷第11期
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doi: 10.3969/j.issn.1000-2561.2025.11.016
  • 接收时间:2025-06-28
  • 首发时间:2026-06-24
  • 出版时间:2025-11-25
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  • 收稿日期:2025-06-28
  • 录用日期:2025-08-05
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广西科技基地和人才专项(桂科AD24010045)
国家自然科学基金项目(32272784; 32160732)
作者信息
    1.广西大学,广西南宁 530004
    2.广西壮族自治区农业科学院,广西南宁 530007
    3.广西果蔬贮藏与加工新技术重点实验室,广西南宁 530007

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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