Isolation of a Major Host Plant-Terpenoid and Its Probable
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Journal of Entomology and Zoology Studies 2016; 4(4): 1351-1358 E-ISSN: 2320-7078 P-ISSN: 2349-6800 Isolation of a major host plant-terpenoid and its JEZS 2016; 4(4): 1351-1358 © 2016 JEZS probable impact on the degenerative growth of Received: 23-05-2016 Accepted: 24-06-2016 female Indian lac insect, Kerria lacca (Kerr.) Sondipon Chakraborty Animal Behavior and Natural Product Research Laboratory, Department of Zoology, West Sondipon Chakraborty, Ankur Chaudhuri, Tanmoy Dutta, Joydip Ghosh, Bengal State University, Berunanpukuria, Mallikapur, Barasat, North 24 Parganas, Samir Kumar Saha, Suman Biswas, Sibani Chakraborty, Soumen Kolkata, West Bengal, India. Bhattacharjee, Chiranjib Pal, Ipsit Chakrabarti, Hemanta Chowdhury Ankur Chaudhuri Department of Microbiology, West Bengal State University, Berunanpukuria, Mallikapur, and Narayan Ghorai Barasat, North 24 Parganas, Kolkata, West Bengal, India. Abstract Tanmoy Dutta Cell and Molecular Biology Laboratory, The present work has been attempted to work out the isolation and characterization of a terpenoid from a Department of Zoology, University of North major host plant Butea monosperma of Indian lac insect (Kerria lacca), and its probable impact on the Bengal, Raja Rammohunpur, Darjeeling, West Bengal, India. degenerative growth pattern of the female morph, which losses its legs and antennae in its adult stage. The studied terpenoid (C30H52O) as ingested from the host plant by the female K. lacca probably plays a Joydip Ghosh Molecular Immunology and Therapeutics key role by inhibiting the acetylcholinesterase (AChE, EC 3.1.1.7) undergoing the process of Laboratory, Department of Zoology, West Bengal State University, Berunanpukuria, uncompetitive inhibition. This type of inhibition is supported by bio-informatics studies also. This Mallikapur, Barasat, North 24 Parganas, inhibition of AChE may be one of the responsible factors for the degenerative growth pattern of female Kolkata, West Bengal, India. K. lacca featuring the loss of all locomotory organs (legs) and antenna, etc. This is most probably caused Samir Kumar Saha by the rapid deacceleration of the molar hydrolysis of substrate acetylcholine by the enzyme AChE under Animal Behavior and Natural Product Research Laboratory, Department of Zoology, West such uncompetitive inhibition by the terpenoid as studied. Bengal State University, Berunanpukuria, Mallikapur, Barasat, North 24 Parganas, Kolkata, West Bengal, India. Keywords: Butea monosperma, Kerria lacca, terpenoid, acetylcholinesterase, uncompetitive inhibition Suman Biswas Department of Chemistry, West Bengal State University, Berunanpukuria, Mallikapur, Introduction Barasat, North 24 Parganas, Kolkata, West Most intriguing domain of insect-plant interactions is selection of plants as host, and Bengal, India. adaptation of insects against the detrimental secondary plant metabolites on which they thrive Sibani Chakraborty [1, 2] Department of Microbiology, West Bengal State . Host plants produce several secondary metabolites (alkaloids, phenolics and terpenoids) University, Berunanpukuria, Mallikapur, [3] Barasat, North 24 Parganas, Kolkata, West to resist against their herbivorous pests . Few plant secondary metabolites, such as Bengal, India. allelochemicals, have been known to inhibit the growth parameters of its target pest organisms Soumen Bhattacharjee by several ways, such as by inhibition of neurotransmitters, ion channel receptors and Cell and Molecular Biology Laboratory, [4-9] Department of Zoology, University of North respiration pathway, etc. Insect defense strategies against the plant secondary metabolites Bengal, Raja Rammohunpur, Darjeeling, West in their food items are also predisposed by several detoxifying enzymes belonging to Bengal, India. [10, 11] Cytochrome P450 family . In the present study the Indian lac insect Kerria lacca, has Chiranjib Pal Molecular Immunology and Therapeutics been selected as a model insect to understand the effect of host plant terpenoids on its growth Laboratory, Department of Zoology, West Bengal State University, Berunanpukuria, pattern, especially of the female morph. Indian lac insect, K. lacca belongs to the Order- Mallikapur, Barasat, North 24 Parganas, Hemiptera, Family- Kerriidae. Insects of this order are characterized by modified mouth parts Kolkata, West Bengal, India. known as rostrum. The mouth parts are modified typically for piercing and sucking the plant Ipsit Chakrabarti [12] Dinabandhu Andrews College, Department of tissue-sap . The crawling nymphal stages of these insect, within a few hours of its Zoology, Raja S.C. Mallick Road, Garia, Kolkata, West Bengal, India. emergences from the encrustation shell of mature female lac insects, are able to successfully penetrate the host plant tissues in order to access the phloem tissue sap [13]. Unlike other Hemanta Chowdhury [14] Central Research Institute of Jute and Allied Homopteran, the lac insect, once inserts its proboscis, remains sedentary for the rest of life . Fibers (CRIJAF), Nilgunj, Barasat, North 24 [12] Parganas, Kolkata, West Bengal, India. During this phase the insect starts secreting lac, the only known resin of animal origin . Narayan Ghorai Encrustation of lac resin is essential to minimize their chances of predations since they have a Animal Behavior and Natural Product Research very vulnerable sedentary life [14, 15]. Among different species of host plants, the large scale Laboratory, Department of Zoology, West Bengal State University, Berunanpukuria, production of lac in West Bengal, India, depends mostly on three plants viz. Butea Mallikapur, Barasat, North 24 Parganas, [12, 14, 16] Kolkata, West Bengal, India. monosperma, Ziziphus mauritiana and Schlischeria oleosa . Like all hemi-metabolous Correspondence insect species, K. lacca passes through different nymphal stages during the courses of its post- Narayan Ghorai embryonic development [14]. In early nymphal stages it is ovate in out-line, slightly pointed Animal Behavior and Natural Product Research Laboratory, Department of Zoology, West posteriorly as well as soft, crimson colored body with one pair of antennae and three pairs of Bengal State University, Berunanpukuria, [14] Mallikapur, Barasat, North 24 Parganas, thoracic legs . Kolkata-126, West Bengal, India. ~ 1351 ~ Journal of Entomology and Zoology Studies But in the case of female morph during its post-embryonic Preparation of plant extract and purification and development it loses all types of effective organs, like legs, isolation of single compound from a terpenoids antennae, wings, etc. and becomes pyriform to globular in moiety shape, which is not a usual trait in case of adult males [14]. Isolated bark sample from host plant B. monosperma was air Several terpenoids of host plants are known to be potent dried and crushed with mixture grinder. Solvent chloroform inhibitors of Acetylcholinesterase (AChE) of their was used for the purpose of phytochemical extraction in pestiferous organisms thriving on them [9]. The principal soxhlator. Soxhlated phytochemical was concentrated from objectives of the present study are isolation, characterization, solvent CHCl3 using the rotary evaporator (Buchi, model no. and to ascertain the role of host plant terpenoids on the R3). Column chromatography (CHCl3: Ethyl Acetate = 8: 1) retrogressive growth of female morph of K. lacca, if any. was performed using Silica gel (mesh, 230-400) to purify it. Activated charcoal was used to remove the necessary Materials and Methods pigments from column derived products. Targeted Study periods phytochemicals were fully purified after performing the The entire study was conducted between the months of preparative thin layer chromatography (Silica Gel- GF 254,). March, 2013 to December, 2015. Bark of the host plant was Three different solvent systems had been used at preparative collected after sixth months from the date of pruning of host thin layer chromatography by following the same method, plants (pruning of plant twigs is a common practice in lac viz. Chloroform: Petroleum-Ether (5: 1), Chloroform: Di- culture in India to make the host plants receptive to the K. Ethyl Ether: Pet Ether (10:2:1) and Chloroform: Ethyl lacca nymphs) in order to isolate and purify the inhibitor Acetate (8: 1) respectively. terpenoid molecule of AChE. Structure analysis of purified phytochemicals Study area Qualitative analysis Necessary field works and sample collections were carried Necessary qualitative analysis of alkaloids, phenolics and out at two different Brood lac Culture Farms, Gananondo terpenoids were done following the protocol of Pushkar et al. Saraswati Ashram, Dalpur (23⁰20 ̍17.19 ̎N, 86⁰54 ̍32.44 ̎E) [12]. Block-Chhatna and Tarkajor Brood Lac Culture Farm, Sunukpahari (23⁰09 ̍25.33 ̎N, 86⁰59 ̍01.04 ̎E) Block-Bankura GC-MS Analysis I, at district Bankura, West Bengal, India. TG-5MS (5% Phenyl Methylpolysiloxae) column (30 m length, 0.25 mm diameter and 0.25 μm film thickness) was Study specimen used for gas chromatography with the Helium (He) gas (flow Both nymphal, and adult female morphs of Kerria lacca rate 1ml/min) as mobile phase (Thermo Scientific, model were collected from host plant B. monosperma in order to name- Trace 1300/Gas Chromatography). Instrumental study the specific activity of AChE following the protocol of specification of mass spectrophotometer was Thermo Ellman et al [17]. Scientific Trace 1300 GC equipped with ISQ MS and an In order to have a standard Michel-Mention (MM) plot and AI/AS 1310 Auto-sampler. During