A Plant-Based Extract Mixture for Controlling Spodoptera Litura

Yooboon et al. Chem. Biol. Technol. Agric. (2019) 6:5 https://doi.org/10.1186/s40538-019-0143-6 RESEARCH Open Access A plant-based extract mixture for controlling Spodoptera litura (Lepidoptera: Noctuidae) Thitaree Yooboon1, Anchulee Pengsook2, Atcharee Ratwatthananon2, Wanchai Pluempanupat2* and Vasakorn Bullangpoti1* Abstract Background: Spodoptera litura larvae are polyphagous insects that have become a signifcant pest in recent years. The spread of this pest has led to the continuous usage of insecticides on crops. Some plant extracts have been used as a mixture to control insect pests and improve productivity. Methods: A plant-based mixture was mixed at a ratio of 1:1 v/v to demonstrate the efect on contact toxicity, feed- ing (no-choice test), and enzyme activities on S. litura. The active compounds of P. retrofractum and A. calamus were isolated by preparative thin-layer chromatography (PTLC). Results: Our results showed that binary mixtures from P. retrofractum and A. calamus exhibit the highest contact toxicity and antifeedant activity at a 1:1 ratio of LD30:LD10 dose (3.213 µg/larva P. retrofractum 3.294 µg/larva A. calamus). The main active ingredient from each crude extract was (2E,4E,14Z)-N-isobutylicosa-2,4,14-trienamide+ from P. retrofractum, and beta-asarone and alpha-asarone from A. calamus. Additionally, A. calamus seems to be the synergistic compound. Some compound mixtures increased the glutathione-S-transferase activities in vivo; whereas, almost no signifcant diferences in esterase activities were noted. Conclusion: The results indicated that the ethanolic crude extracts of P. retrofractum and A. calamus mixtures could be used as the pesticidal compound and to develop a binary mixture formulation for controlling lepidopteran pests. However, the toxicity of this mixture to mammals needed to be explored before commercial development. Keywords: Spodoptera litura, Plant extract mixture, Synergist, Detoxifcation enzymes, Antifeedant Introduction environment and people’s health [5]. Tese results of Spodoptera litura (Lepidoptera: Noctuidae) is a poly- insecticide usage have encouraged scientists to seek less phagous insect pest that feeds on at least 87 plant spe- hazardous chemicals and identify an alternative method cies in over 40 plant families, including many vegetables, for integrated pest management (IPM). fruit, cotton, groundnut, chili, tobacco, castor, lady fn- Botanical extract products have become more promi- ger, caulifower, and pulses, in many Asian countries, nent in assessments of current and future pest control such as Tailand, China, Japan, India [1–3]. It has gradu- alternatives. Tey are biodegradable and ecologically ally become a signifcant insect pest in recent years [4]. safe and an important component of IPM programs [6]. Te spread of this pest has led to the continuous usage For instance, numerous studies have focused on neem of insecticides on crops. Pesticide residues impact the (Azadirachta indica A. Juss), and a number of studies demonstrate efcacy against a variety of pests [7, 8]. Its *Correspondence: [email protected]; [email protected] compound has numerous activities against insects, such 1 Animal Toxicology and Physiology Specialty Research Unit, Department as antifeedant, growth inhibition, growth regulation, of Zoology, Faculty of Science, Kasetsart University, Phahonyothin Road, reduced fecundity and sterility, and inhibition of pro- Bangkok 10900, Thailand 2 Department of Chemistry and Center of Excellence for Innovation tein synthesis, as well as toxic efects in a wide variety of in Chemistry, Faculty of Science, and Special Research Unit for Advanced insect taxa, including Lepidoptera [7, 8]. Plants comprise Magnetic Resonance, Kasetsart University, Bangkok 10900, Thailand a source of novel chemical compounds that are used in © The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creat iveco mmons .org/licen ses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creat iveco mmons .org/ publi cdoma in/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Yooboon et al. Chem. Biol. Technol. Agric. (2019) 6:5 Page 2 of 10 medicine and other applications. Each plant contains or only one crude extract [12, 13]. Tus, this research many active compounds, such as terpenes, terpenoids has the main goal of producing a plant-based mixture for alkaloids, steroids, phenols, and favonoids [9], which are control of S. litura with the possibility of increasing the found in specifc parts, i.e., leaves, fowers, stems, fruits, control efciency. Additionally, detoxifcation enzyme seeds, roots. Plant defense compounds typically result activities on treated S. litura were analyzed to search for from a combination of these plant products and not as the possibility of controlling this pest and examining the individual compounds. trends of resistance to this plant-based product in the A variety of documents suggest that complex mixtures future. would be more efcient, and synergistic efects have been reported [10, 11]. Singh et al. [12] demonstrated that Materials and methods thymol and α-terpineol synergized the impacts of both Insect rearing linalool and 1,8-cineole, but linalool with 1,8-cineole Spodoptera litura larvae used in this study were obtained exhibited only an additive efect against Chilo partellus. from a laboratory colony maintained in the Animal trans-Anethole acted synergistically with thymol, citron- Toxicology and Physiology Specialty Research Unit ellal and α-terpineol [13]. Consequently, the mixtures (ATPSRU), Department of Zoology, Faculty of Science, of plant compounds are also likely to be more durable Kasetsart University. Te culture was continuously main- against insects evolving resistance and developing behav- tained on an artifcial diet (mixture of 240 g of green ioral desensitization. bean, 25 g of agar, 40 ml of mixed vitamin solution, 5 g Tis study was focused on the efect of six plant of ascorbic acid, 40 ml of amoxicillin solution, 3 g of sor- extracts, including Acorus calamus (Acorus calamus: bic acid, 5 g of methylparaben, 20 g of yeast, 4 ml of 40% Acorus calamus), Alpinia galanga (Zingiberaceae: Zin- formalin and 1.41 l of water) in the insect-rearing room giberales), Curcuma longa (Zingiberaceae: Zingiberales), of the Department of Zoology, Faculty of Science, Kaset- Piper nigrum (Piperaceae: Piperales), Piper retrofrac- sart University, at 26 °C with 75% RH and a 16:8-h L:D tum (Piperaceae: Piperales), and Sphagneticola trilobata photoperiod. Second and third instar larvae were used (Asteraceae: Asterales), on S. litura control. Tese plant randomly for the treatment. All experimental proce- species are well known for their natural properties in the dures in this research were performed with the approval Tai traditional system of medicine or growth as a weed of an appropriate animal Ethics Committee of Kaset- and have been reported to possess numerous types of sart University, Tailand, under the reference number biological activities. OACKU01059. Numerous anti-insect properties of the six plants involve toxic efects against many insects. Lee et al. [14] Plant materials and extraction methods demonstrated the insecticidal efect of the methanol Te rhizomes of A. galanga, C. longa, and A. calamus; the extract of C. longa rhizome on Plutella xylostella lar- leaves and stem of S. trilobata; and the fruits of P. nigrum vae. Te hexane and ethanol extracts of rhizomes of A. and P. retrofractum were obtained from Banphoromyen, galanga exhibit insecticidal activity against the fruit fy Amphawa, Samut Songkhram province, Tailand. Each Bactrocera dorsalis when applied using a direct spray plant was rinsed with water to remove debris and air technique [15]. Te essential rhizome oil of A. calamus dried under shade. Dried plants were chopped fnely showed a sterilizing efect against the eggs of Sitophilus to a powder. One kilogram of each powder sample was granarius, Sitophilus oryzae, and Culiosobruchus chinen- soaked in ethanol for 14 days. Each crude extract was fl- sis [16] and larvicidal activity against Culex quinquefas- tered using a vacuum pump, dried by a rotary evaporator ciatus [17]. Upadhyay and Jaiswal found that 0.2 μl of P. to obtain the solidifed crude extracts and stored at 4 °C nigrum oil signifcantly repelled Tribolium castaneum in a refrigerator until further processing. [18]. Te dichloromethane extract of P. nigrum has pesticidal activity against Callosobruchus maculates and Preliminary test of the contact toxicity bioassay for crude Sitophilus zeamais [19]. In addition to P. nigrum, P. ret- extract rofractum, which is in the same genus, also showed larvi- Contact toxicity bioassays were performed with second cidal activity against mosquito larvae C. quinquefasciatus instar larvae of S. litura. Each ethanolic crude extract [20]. Moreover, S. trilobata crude extracts have a larvi- was evaluated individually to determine efcacy levels cidal efect on S. litura, S. exigua, and P. xylostella larvae upon topical application to the thorax region with vari- after topical application [21]. ous concentrations of extracts (2–140 µg/larva) using However, there is no research on the extract

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