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本课题组参照文献[1]采用组织分离法对火龙果果腐病病原菌进行分离纯化,并通过形态学及分子生物学对病原菌进行鉴定,确定引起火龙果果腐病的病原菌为桃吉尔霉(Gilbertella persicaria)。植物精油是一类植物源次生代谢物质,分子量较小,且具有一定挥发性[2],具有对环境友好的优势。植物精油在抑菌方面和果蔬采后保鲜方面均表现出较好的效果[3]。本实验研究植物精油对火龙果果腐病病原菌的抑菌效果,为后期使用天然植物精油对火龙果果腐病进行防治提供技术支撑。
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桃吉尔霉(G. persicaria)编号为HLG-T,分离自海南省东方市某果园的贮藏期病果。由中国热带农业科学院环境与植物保护研究所采后病害及贮运保鲜课题组分离鉴定,经致病性测定后纯化保存备用。供试植物精油:牛至精油、香茅精油、肉桂精油、芥末精油、丁香精油、樟脑精油、玫瑰草精油、草果精油、山茶花精油、茶树精油、柠檬精油和橄榄精油等12种精油。上述精油均购自上海源叶生物科技有限公司。接种用实验果实采购自海南省海口市南北水果市场,选取新鲜健康、表面无损伤、大小和成熟度一致的果实。
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设置12种植物精油的不同浓度梯度,以无菌水为空白对照。植物精油的抑菌效果实验采用含药平板法[4]。将病原菌菌饼(直径d=5 mm)接种于PDA平板中央,置于28 ℃恒温培养箱中培养24 h后,采用十字交叉法测量菌落直径,每处理3次重复。依据公式(1)计算抑菌率,以质量浓度的对数值为横坐标,以抑菌率几率值为纵坐标求毒力回归方程,计算相关系数、EC50值,比较供试12种植物精油对供试菌株的毒力大小。
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病害控制效果实验采用浸泡法。参照缪应林[5]等方法稍作修改,选用抑菌效果较好的5种植物精油,配制成1 000 μL·L−1精油溶液,将打好孔(d=5 mm)的火龙果果实在1 000 μL·L−1精油溶液中浸泡10 min,晾干后接种病原菌菌饼(d=5 mm),装入密封保鲜盒中,于25 ℃恒温保湿培养。每处理设置3次重复,以无菌水处理为空白对照。2 d后采用“十字交叉法”测量病斑直径,按公式(2)进行计算。
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在供试的12种植物精油中,牛至精油的抑菌效果最好,其EC50值仅为94.1 μg·mL−1;其次是香茅精油、肉桂精油、芥末精油、丁香精油、樟脑精油、玫瑰草精油和草果精油,EC50值依次为122.9,174.0,254.3,261.1,311.6,349.6,372.7 μg·mL−1,均在100.0~500.0 μg·mL−1;而山茶花精油、茶树精油、柠檬精油、橄榄精油的抑菌效果不好,EC50值均在500.0 ~ 1000.0 μg·mL−1。
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在供试的5种精油中,芥末精油对该病害的控制效果最好,其控制率达到100%;其次是牛至精油和丁香精油,其控制率达到91.96%和56.80%;香茅精油和肉桂精油抑菌效果最差,其控制率均在50%以下。牛至精油对该病害的控制率虽然达到91.96%,但使用牛至精油浸果极易对果面造成药害。
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植物精油是从植物组织中提取的物质,具有抑菌广谱、绿色安全、高效长效和不易产生抗性等特点,是一种很有潜力的新型杀菌物质[6-7]。供试12种精油对于桃吉尔霉(G. persicaria)在离体条件下的生长均有抑制作用,其中牛至精油抑菌效果最好,其次是香茅精油、肉桂精油、芥末精油、丁香精油、樟脑精油、玫瑰草精油和草果精油的抑菌效果也较强。选取抑菌效果较好的5种精油(牛至精油、香茅精油、肉桂精油、芥末精油、丁香精油)进行浸果实验,病害控制结果表明,芥末精油对该病害的控制效果最好,其控制率达到100%,且果面未见损伤;牛至精油其次,其控制率虽然达到91.96%,但是牛至精油处理使果实出现严重药害,不适合在实际生产中应用。综上所述,对桃吉尔霉(G. persicaria)引起的火龙果果腐病控制效果最好的是芥末精油,该精油处理可抑制该病原菌的生长,减少该菌引起的火龙果果腐病的发生。
Screening of Plant Essential Oils with Antifungal Activities Against Pitaya Fruit Rot
doi: 10.15886/j.cnki.rdswxb.2021.01.010
- Received Date: 2020-09-18
- Rev Recd Date: 2020-12-03
- Available Online: 2021-04-08
- Publish Date: 2021-04-12
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Key words:
- pitaya /
- fruit rot /
- Gilbertella persicaria /
- plant essential oil /
- dipping treatment
Abstract: Twelve plant essential oils were tested to observe their inhibitory effect on Gilbertella persicaria that causes fruit rot of pitaya or dragon fruit in order to clarify the sensitivity of G.persicaria to plant essential oils during the storage of pitaya. The results showed that all the tested 12 plant essential oils had an inhibitory effect on G. persicaria. The top 5 plant essential oils with higher antifungal activities are oregano essential oil, citronella essential oil, cinnamon essential oil, mustard essential oil and clove essential oil, which had the EC50 values of 94.1, 122.9, 174.0, 254.3 and 261.1 μg·mL−1, respectively. Pitaya fruits were treated with these 5 essential oils at a concentration of 1 000 μL·L−1 by dipping, and inoculated with the fungi G.persicaria to observe the control effect of the essential oils on the fungi. The results showed the mustard essential oil had the control rate of 100% against G. persicaria and caused no damage to the surface of the fruit, while the oregano essential oil had a control rate of 91.96% against G. persicaria, but caused serious damage to the fruit surface, which is not suitable for application in production.
Citation: | YU Jing, LI Min, GAO Zhaoyin, GONG Deqiang, ZHANG Shaogang, HONG Xiaoyu, HUA Jingjing, HU Meijiao. Screening of Plant Essential Oils with Antifungal Activities Against Pitaya Fruit Rot[J]. Journal of Tropical Biology, 2021, 12(1): 72-74. doi: 10.15886/j.cnki.rdswxb.2021.01.010 |