摘要
将光动力抗菌化学疗法(Photodynamic antibiotic therapy,PACT)引入植物病害防治,研究了叶绿素降解产物二氢卟吩e6(Ce6)对尖孢镰刀菌的光敏抑制作用。通过化学方法以蚕砂叶绿素为原料,在浓碱溶液中加热水解后酸化,合成叶绿素降解产物二氢卟吩e6,通过红外光谱(IR)、高效液相色谱(HPLC)等对合成产物进行结构表征。选用植物枯萎病的主要病原菌——尖孢镰刀菌(Fusarium oxysporum)进行离体抑菌活性试验,采用菌丝生长速率法和凹玻片悬滴法分别测定Ce6在光照和黑暗条件下对尖孢镰刀菌菌丝径向生长和分生孢子萌发的抑制作用,并进行比较与分析。试验结果:二氢卟吩e6可通过蚕砂叶绿素碱水解后再酸化来合成,合成产物纯度可达62.6%。Ce6供试药液对尖孢镰刀菌菌丝生长的抑制作用随药液浓度的增大而增大,且有无光照对其抑菌效果有一定影响;20 mg/m L供试药液对菌丝生长的抑制率为41.60%,在黑暗中的抑制率为25.86%;10 mg/m L供试药液在光照下对菌丝生长的抑制作用为38.78%,在黑暗条件抑制率为14.28%,高浓度(≥10 m L)与低浓度(≤5 m L)之间差异显著(P<0.05),光照条件下其EC50为35.302 mg/m L。试验结果还表明:光照条件下,Ce6能有效抑制尖孢镰刀菌分生孢子的萌发,抑制率随药液浓度的增大而增大,且其在高浓度时对光照不敏感(P>0.05),而在低浓度时较为敏感(P<0.05);40 mg/m L供试药液在光照下对分生孢子的抑制作用可达94.92%,在黑暗中其抑制率也可达到85.28%,均表现出了良好的抑制作用;低浓度药液在光照和黑暗条件下对分生孢子的抑制作用表现出了较大的差异(P<0.05),光照条件下Ce6供试药液EC50为2.713 mg/m L,黑暗条件下其EC50为14.035 mg/m L,前者远小于后者,说明药液的抑制作用光照下使用明显优于避光使用。
Photodynamic antibiotic therapy（ PACT） was introduced into the prevention and control of plant diseases,and then the photosensitizing effect of chlorin e6 on Fusarium oxysporum was explored Chlorin e6 was prepared by a chemical method,using silkworm excrement chlorophyll as raw materials,and then characterized by infrared spectrum analysis（ IR） and high performance liquid chromatography（HPLC）.The possible mechanisms about corresponding reactions were tentatively proposed.Fusarium oxysporum,the main pathogen of plant wilt,was used to determine its in vitro antifungal activity,using mycelial growth rate and hanging drop method.Ce6 could be synthesized by alkali hydrolysis and then acidification from silkworm chlorophyⅡand the purity reached 62.6%.The inhibitory effect of chlorin e6 test solution on mycelial growth of F.oxysporum increased with concentration,and light had a certain effect on the antifungal effect.The inhibitory rate of20 mg/mL test solution on mycelial growth was 41.60%under light condition and 25.86%in darkness.And the inhibitory effect of 10 mg/mL test solution was 38.78%under light condition and14.28%in darkness.There was a significant difference between high concentration（≥10 mL）and low concentration（≤5 mL）（P〈0.05）,and the35.302 mg/mL under light condition.The results of inhibition of spore germination showed that Ce6 could effectively inhibit the germination of F.oxysporum conidia under light condition,and the inhibition rate increased with the increase of concentration,and it was not sensitive to light at high concentration（P〈0.05）,but sensitive at lower concentration（P〈0.05）.The inhibitory effect of 40 mg/mL test solution on conidia under light condition was up to 94.92%,and the inhibitory rate in darkness could reach 85.28%,both showing a good inhibition.However,the inhibitory effect of lower concentration solution on conidia under light and dark conditions showed a significant difference（P〈0.05）.TheCe6test solution was 2.713 mg/mL and 14.035 mg/mL under light and dark conditions,respectively,indicating that the inhibitory effect under light condition was obviously better than that of darkness.
作者
杨璟
张国财
胡春平
林连男
蔡良苑
YANG Jing;ZHANG Guocai;HU Chunping;LIN Liannan;CAI Liangyuan(School of Forestry, Northeast Forestry University, Harbin 150040, China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2018年第3期270-275,共6页
Journal of Jilin Agricultural University
基金
中央高校基本科研业务费专项资金项目(2572016AA03
2572017AA18)
哈尔滨市应用技术研究与开发项目(2016RAXXJ035)
国家级大学生创新创业训练计划项目(201710225165)
关键词
二氢卟吩e6
尖孢镰刀菌
光敏抑菌活性
光活化农药
chlorin e6
Fusarium oxysporum
photosensitive antifungal activity
photoactivated fungicide
