摘要
【目的】探究植物次生代谢物质丁香酚对美国白蛾幼虫的毒杀和拒食活性,明晰丁香酚对幼虫生长发育、取食消化和氧化防御系统的作用机制。【方法】通过生物测定、体内酶活性试验和实时荧光定量PCR(RTqPCR)法,分析了丁香酚对美国白蛾3龄幼虫的毒杀效果计算得到亚致死浓度(LC_(30)),以LC_(30)的丁香酚处理幼虫,测定其体重、取食量、食物利用率、胰蛋白酶(TYP)、α-淀粉酶(α-AMS)、脂肪酶(LPS)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的活性以及过氧化氢(H_(2)O_(2))含量、丙二醛(MDA)含量和总抗氧能力(T-AOC),并在mRNA水平上分析了上述消化酶和抗氧化酶基因的表达水平。【结果】1)在最高浓度39.408 mg/mL的丁香酚处理72 h后,幼虫校正死亡率达到66.70%。丁香酚处理幼虫72 h后的致死中浓度(LC50)为21.001 mg/mL,亚致死浓度(LC_(30))为9.998 mg/mL;2)在LC_(30)的丁香酚处理72 h后,幼虫取食量下降15.10%(P<0.05),体质量增长量下降了32.28%,食物利用率(ECI)下降20.38%;3)LC_(30)的丁香酚处理72 h后,幼虫体内TYP、α-AMS和LPS活性均显著下降,相关基因Hcα-AMS1、Hcα-AMS2、HcLPS1、HcLPS2、HcTYP1和HcTYP2也均显著下调;4)幼虫体内氧化应激相关指标中H_(2)O_(2)含量增加至对照组的2.93倍、MDA含量提升至对照组的1.91倍、T-AOC则下降11.55%。抗氧化酶CAT、SOD和POD酶活性均显著上升,相关基因HcCAT1、HcCAT2、HcSOD1、HcSOD2、HcPOD1和HcPOD2的表达量也都显著上调。【结论】丁香酚对美国白蛾3龄幼虫具有良好的毒杀和拒食作用。丁香酚干扰幼虫体内消化吸收系统和氧化防御系统,导致幼虫产生氧化损伤、取食量减少和消化功能受阻等一系列不利于其生存现象,最终导致其死亡。
【Objective】To investigate the toxicological and antifeedant activity of eugenol,a plant secondary metabolite,on Hyphantria cunea larvae,and to clarify the mechanism of action of eugenol on larval growth and development,feeding and digestion,and oxidative defense system.【Method】The toxic effect of eugenol on 3rd instar larvae of H.cunea was analyzed by bioassay,in vivo enzyme activity assay and real-time fluorescence quantitative PCR(RT-qPCR)method to calculate the sublethal concentration(LC_(30)),and the larvae were treated with eugenol at LC_(30),and their body weights,feed intake,food utilization,trypsin(TYP),α-amylase(α-AMS),lipase(LPS),superoxide dismutase(SOD),catalase(CAT)and peroxidase(POD)activities as well as hydrogen peroxide(H_(2)O_(2))content,malondialdehyde(MDA)content and total antioxidant capacity(T-AOC),and the expression levels of the above mentioned digestive and antioxidant enzyme genes were analyzed at mRNA level.【Result】1)The corrective mortality rate of larvae reached 66.70%after 72 h of treatment with the highest concentration of 39.408 mg/mL eugenol.The lethal median concentration(LC50)of eugenol-treated larvae after 72 h was 21.001 mg/mL and the sublethal concentration(LC_(30))was 9.998 mg/mL;2)After 72 h of eugenol treatment at LC_(30) post-larval feeding was reduced by 15.10%,body weight gain was reduced by 32.28%,and food utilization(ECI)was reduced by 20.38%under eugenol treatment at LC_(30);3)TYP,α-AMS and LPS activities were significantly decreased in larvae after 72 h of eugenol treatment at LC_(30),and the related genes,Hcα-AMS1,Hcα-AMS2,HcLPS1,HcLPS2,HcTYP1,and HcTYP2,were also significantly down-regulated;4)Among the oxidative stress-related indicators in larvae,the H_(2)O_(2) content increased to 2.93 times of the control group,the MDA content increased to 1.91 times of the control group,and the T-AOC decreased by 11.55%.The activities of antioxidant enzymes CAT,SOD and POD were significantly increased,and the expressions of related genes HcCAT1,HcCAT2,HcSOD1,HcSOD2,HcPOD1 and HcPOD2 were also significantly up-regulated.【Conclusion】Eugenol has good toxicity and food rejection effects on the 3rd instar larvae of H.cunea.Eugenol interferes with the digestive and absorptive systems and oxidative defence system of the larvae,resulting in oxidative damage,reduced feeding and impaired digestion,which are detrimental to the survival of the larvae and ultimately lead to their death.
作者
谢江艳
王亚军
孙妍
李奇萌
孙岩圣
崔书铭
张国财
毕冰
XIE Jiangyan;WANG Yajun;SUN Yan;LI Qimeng;SUN Yansheng;CUI Shuming;ZHANG Guocai;BI Bing(School of Forestry,Northeast Forestry University,Harbin 150040,Heilongjiang,China;Heilongjiang Forest Protection Research Institute,Harbin 150040,Heilongjiang,China)
出处
《中南林业科技大学学报》
北大核心
2025年第11期87-96,共10页
Journal of Central South University of Forestry & Technology
基金
中央财政林业科技推广示范项目(黑〔2023〕TG21号)
黑龙江省科技厅应用研究项目(LJGXCG2022-010)
内蒙古大兴安岭重点国有林管理局(森工集团)林业科技项目(2018003)。
关键词
丁香酚
美国白蛾
生物测定
消化代谢
氧化应激
氧化还原酶
eugenol
Hyphantria cunea
bioassay
digestive metabolism
oxidative stress
oxidoreductase
