诱集植物香根草对二化螟幼虫致死的作用机制

doi:10.3864/j.issn.0578-1752.2017.03.007

摘要/Abstract

摘要： 【目的】香根草(Vetiveria zizanioides)能够有效诱集水稻害虫二化螟（Chilo suppressalis）雌成虫在其上产卵，但孵化出的幼虫在香根草上不能完成生活史。本研究旨在明确香根草对二化螟幼虫致死的作用机制，为开发以香根草为基础的水稻螟虫可持续控制新技术提供理论依据。【方法】通过有机溶剂萃取和硅胶柱层析方法提取香根草有毒活性物质，并利用饲料混毒法测定各提取组分对二化螟3龄幼虫的生物活性，同时利用生物化学方法测定水稻和香根草中营养物质含量以及二化螟3龄幼虫取食水稻和香根草后体内相关解毒酶和消化酶的活性变化。【结果】香根草石油醚萃取物处理二化螟3龄幼虫9、12、15 d后，死亡率分别为38.89%、57.41%、85.19%，显著高于乙酸乙酯萃取物、正丁醇萃取物等其他提取组分处理二化螟幼虫后的死亡率。进一步从香根草石油醚萃取物中分离获得组分1（Fr1）和组分5（Fr5）,对二化螟3龄幼虫的具有较高毒性，以0.05 g·mL^-1浓度混入人工饲料饲喂3 d后死亡率分别为85.00%和67.67%，说明香根草中含有对二化螟幼虫具有致死作用的有毒活性物质。香根草中总蛋白、纤维素、总糖、氨基酸等营养物质含量分别为9.45 μg、1.61%、1.45%、4.00%，均显著低于水稻中相应营养物质含量，其中水稻中蛋氨酸含量是香根草中的7.0倍。而香根草中单宁含量则显著高于水稻中的含量，是水稻中的1.31倍。此外，二化螟取食水稻和香根草3 d后，取食香根草的3龄幼虫体内蛋白酶、淀粉酶、海藻糖酶和蔗糖酶等消化酶的活性显著低于取食水稻幼虫体内相应消化酶的活性；6 d后，取食香根草的3龄幼虫体内细胞色素P450酶活性显著低于取食水稻的幼虫；9 d后，取食香根草的3龄幼虫体内羧酸酯酶CarE的活性显著低于取食水稻的幼虫。与取食水稻的3龄幼虫相比，取食香根草的幼虫体内谷胱甘肽硫转移酶GSTs的活性也有所降低，但是两者在统计学上没有显著差异。【结论】香根草作为二化螟的致死型诱集植物，对二化螟幼虫的致死作用主要表现在两方面：一是香根草中含有对二化螟幼虫具有致死作用的有毒活性物质，这些物质通过抑制幼虫体内解毒酶CarE和P450酶的活性，使幼虫逐渐丧失解毒代谢能力，最终死亡；二是香根草相对水稻营养物质匮乏，二化螟幼虫取食香根草后营养不均衡，从而影响体内消化酶活性，造成消化功能紊乱，影响其正常生理活动，最终死亡。

关键词: 香根草, 二化螟, 致死作用, 营养物质, 解毒酶, 消化酶

Abstract: 【Objective】 Vetiver grass (Vetiveria zizanioides) can effectively attract female adults of Chilo suppressalis to lay eggs, while the larvae can not complete their life cycles by feeding on vetiver. The objective of this study is to illuminate the lethal mechanism of trap plant-vetiver against the larvae of C. suppressalis and provide a theoretical basis for the development of sustainable control technology for C. suppressalis based on the application of trap plant vetiver in rice based ecosystem. 【Method】 The toxic substances were extracted from vetiver by the methods of extraction and silica gel column chromatography, and the biological activities of these extracts against 3rd instar larvae of C. suppressalis were determined by mixing them in the artificial diet. The nutrient contents of rice and vetiver, and the detoxifying and digestive enzymes activities of C. suppressalis larvae fed on rice and vetiver were analyzed using biochemical methods. 【Result】 The mortalities of the C. suppressalis 3rd instar larvae after 9, 12, 15 days treated by sherwood oil extract of vetiver were 38.89%, 57.41%, and 85.19%, respectively, which were significantly higher than the mortalities of larvae treated by ethyl acetate extract, n-butyl alcohol extract and other fractions. The fraction 1 (Fr1) and fraction 5 (Fr5) were isolated and obtained from sherwood oil extract of vetiver, and the mortalities of C. suppressalis the 3rd instar larvae to Fr1 and Fr5 (0.05 g·mL^-1) were 85.00% and 67.67% after 3 days treatment, respectively. These results indicated that vetiver contains toxic substances, which have lethal effect on C. suppressalis larvae. Results of nutrient contents determination showed that the contents of total protein, cellulose, total sugar and amino acids in vetiver were 9.45 μg, 1.61%, 1.45%, 4.00%, respectively, significantly lower than those in rice, and the methionine (MET) content in rice was 7-fold higher than that in the vetiver. However, the content of tannin was 1.31-fold higher than that in rice. In addition, the protease, amylase, trehalase and sucrase enzyme activities of C. suppressalis larvae on 3rd day after fed (DAF) on vetiver were significantly lower than those in the larvae fed on rice; and on the 6th DAF, the activity of P450 enzyme in the larvae fed on vetiver was significantly lower than that in the larvae fed on rice; and the esterase activity of C. suppressalis larvae on the 9th DAF on vetiver was significantly lower than that in the larvae fed on rice. Compared to glutathione S-transferase (GSTs) activity of the 3rd instar larvae feeding on rice, the enzyme activity of the larvae feeding on vetiver was also decreased, however, there was no significant difference by statistical analysis. 【Conclusion】 Vetiver as a dead-end trap plant of C. suppressalis, there were probably two lethal mechanisms of vetiver against C. suppressalis. Firstly, vetiver contains toxic substances, which have lethal effect on C. suppressalis and these substances inhibited the esterase and cytochrome P450 enzyme activities, leading to the function loss of larvae in detoxification and metabolism. Secondly, compared to rice, lack of nutrients in vetiver can not meet the demand of the larvae, and led to digestive function and physiological activity disorder and ultimately death.

Key words: vetiver, Chilo suppressalis, lethal effect, nutrient, detoxifying enzyme, digestive enzyme

鲁艳辉，高广春，郑许松，吕仲贤. 诱集植物香根草对二化螟幼虫致死的作用机制[J]. 中国农业科学, 2017, 50(3): 486-495.

LU YanHui, GAO GuangChun, ZHENG XuSong, Lü ZhongXian. The Lethal Mechanism of Trap Plant Vetiveria zizanioides Against the Larvae of Chilo suppressalis[J]. Scientia Agricultura Sinica, 2017, 50(3): 486-495.

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