昆虫学报 ›› 2016, Vol. 59 ›› Issue (8): 839-845.doi: 10.16380/j.kcxb.2016.08.005

• 研究论文 • 上一篇    下一篇

黄皮种子提取物对斜纹夜蛾幼虫的杀虫活性及有效成分鉴定

郭成林1,2, 覃柳燕3, 万树青1,*, 马跃峰2, 覃建林2, 马永林2   

  1. (1. 华南农业大学, 农药与化学生物学教育部重点实验室, 广州 510642; 2. 广西农业科学院植物保护研究所, 广西作物病虫害生物学重点实验室, 南宁 530007; 3. 广西农业科学院生物技术研究所, 南宁 530007)
  • 出版日期:2016-08-20 发布日期:2016-08-20

Insecticidal activity and identification of the active ingredients of Clausena lansium seed extracts against larval Spodoptera litura (Lepidoptera: Noctuidae)

GUO Cheng-Lin 1,2, QIN Liu-Yan3, WAN Shu-Qing1,*, MA Yue-Feng2, QIN Jian-Lin2, MA Yong-Lin2   

  1. (1. Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou 510642, China; 2. Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests, Institute of Plant Protection, Guangxi Academy of Agricultural Sciences, Nanning 530007, China; 3. Institute of Biotechnology, Guangxi Academy of Agricultural Sciences, Nanning 530007, China)
  • Online:2016-08-20 Published:2016-08-20

摘要: 【目的】测定黄皮Clausena lansium种子提取物对斜纹夜蛾Spodoptera litura幼虫的拒食活性,对有效成分进行分离、鉴定,并进一步测定其拒食活性、生长发育抑制活性及细胞毒性,为筛选斜纹夜蛾生态防治植物源杀虫剂提供依据。【方法】采用叶蝶法测定黄皮种子甲醇提取物及其石油醚、乙酸乙酯、正丁醇和水4种溶剂萃取物对斜纹夜蛾3龄幼虫的非选择性拒食活性和生长发育抑制活性;用硅胶柱层析进行石油醚萃取物和乙酸乙酯萃取物拒食活性成分分离;用核磁共振谱(NMR)进行拒食有效成分结构鉴定;用MTT法测定拒食活性成分对SL细胞毒性。【结果】黄皮种子甲醇提取物对斜纹夜蛾3龄幼虫具有较强的拒食活性,活性成分主要分布于石油醚相和乙酸乙酯相中;石油醚相和乙酸乙酯相经硅胶柱层析分离得10个馏分,以黄色晶体馏分2的拒食活性最强;经分离纯化及核磁共振氢谱和碳谱鉴定,黄皮新肉桂酰胺B(lansiumamide B)为主要的拒食活性成分,对斜纹夜蛾3龄幼虫48 h拒食中浓度为214.95 μg/mL,同时具有较强的细胞毒性,对SL细胞48 h抑制中浓度为10.09 μg/mL。【结论】黄皮种子提取物对斜纹夜蛾幼虫具有较强的拒食活性及细胞毒性,黄皮新肉桂酰胺B是主要活性成分之一,作为植物源杀虫剂开发具有较好的发展前景。  

关键词: 斜纹夜蛾, 黄皮, 提取物, 新肉桂酰胺B, 拒食活性, 细胞毒性

Abstract: 【Aim】 By measuring the antifeedant activities of extracts from Clausena lansium seeds against Spodoptera litura larvae as well as separating and identifying the active ingredients and measuring the antifeedant activity, development deterrent activity and cytotoxicity, this study aims to provide references for screening botanical pesticides for ecological management of S. litura. 【Methods】 Leaf disc method was used to determine the no-choice antifeedant activities of methanol extract of C. lansium seeds and extracts of other four solvents (petroleum ether, ethyl acetate, n-butanol and water) as well as the developmental inhibitory activity against the 3rd instar larvae of S. litura. Silica column chromatography was used to separate the active antifeedant ingredients of petroleum ether and ethyl acetate extracts. Nuclear magnetic resonance spectroscopy (NMR) was used to identify the structure of active antifeedant compound, and MTT method was used to determine the cytotoxicity of antifeedant compound to S. litura cell lines. 【Results】 The methanol extract of C. lansium seeds exhibited strong antifeedant activity against the 3rd instar larvae of S. litura. The active ingredients were mainly distributed in petroleum ether extracts and ethyl acetate extracts. Extracts of petroleum ether and ethyl acetate were fractioned with silica column chromatography and 10 fractions were collected then. Fraction 2, a yellow crystal, showed the highest antifeedant activity. Through isolation, purification and 1HNMR and 13CNMR analyses, lansiumamide B was identified to be the main active ingredient, with the 48 h median antifeedant concentration (AFC50) of 214.95 μg/mL against the 3rd instar larvae of S. litura. This ingredient also displayed high cytotoxicity to the 3rd instar larvae of S. litura, with the 48 h cell medium inhibition concentration value of 10.09 μg/mL. 【Conclusion】 Extracts from C. lansium seeds have strong antifeedant activities and cytotoxicity against S. litura larvae. Lansiumamide B is one of the main active ingredients of the extracts, suggesting its good application prospect as a botanical insecticide.    

Key words: Spodoptera litura; Clausena lansium, extract, lansiumamide B, antifeedant activity, cytotoxicity