Larvicidal potential of purified protease inhibitor from Solanum aculeatissimum Jacq. against Odoiporus longicollis Oliver.
Keywords:
Solanum acculeatissimum, Purification, Larvicidal activity, Odoiporus longicollis, Protease inhibitor, MortalityAbstract
Protease inhibitor was isolated from the fruits of Solanum acculeatissimum Jacq. (SAPI) and purified by four steps. PI activity was assayed by estimating the residual hydrolytic activity of trypsin and chymotrypsin towards the substrates BAPNA (N-benzoyl-L-arginine-p-nitroanilide) and BTPNA (N-benzoyl-L-tyrosyl-p-nitroanilide), respectively, at pH 8.0 after pre-incubation with inhibitor. Molecular mass of purified PI was evaluated as 22.2 kDa by SDS–PAGE. Affinity column chromatography yielded a single prominent elution peak. The purity was further checked by RP-HPLC showing a single peak, coinciding with the protein peak. Specific activities of purified PI were 502 TlU and 433.7 CIU U/mg, towards the substrates of trypsin and chymotrypsin respectively with 0.95 mg protein content. Overall, the specific activities increased to 92.6 and 82.9 folds with 9.8 and 8.77 % yield with respect to the substrates. Subsequently, the larvicidal potential of different concentrations of purified SAPI was evaluated against the banana pest Odoiporus longicollis following the treatment on third, fourth and fifth instars larvae. The results of the total percent mortality (from 3rd to 5th instar and adult) demonstrated that the mortality rate progressively increased with the concentrations of SAPI (50 to 250 µM). A minimum of 8% mortality was observed with 50 µM concentration of the PI. It was significant to note that the percent deformity of adults noticed from 50 µM onwards was more than 2%. Thus, the present data suggests an eco-friendly approach for the control of agricultural pest, O. longicollis. Further studies are planned to trace the molecular mechanism of larvicidal potential of SAPI against the pest.
References
. Indira Priyadarshini G, Mukherjee U, Nagendra Kumar. ‘Biology and seasonal incidence of pseudostem weevil, Odoiporus longicollis Olivier (Coleoptera: Curculionidae) in banana.’ Pest Management in Horticultural Ecosystems. 2014; 20(1): 8-13. [2]. Padmanaban B, Sathiamoorthy S. International Network for the improvement of banana and Plantain, Montpellire Cedex. 2001; 5. [3]. Broadway RM, Duffey SS. ‘Plant proteinase inhibitors: mechanism of action and effect on the growth and digestive physiology of larval Heliothis zea and Spodoptera exigua.’ Journal of Insect Physiology.1986; 32: 827-833. [4]. Paulraj KL, Kripa RK. ‘Plant protease inhibitors in control of phytophagous insects’, Electronic Journal of Biotechnology. 2002; 5(1): 93-109. [5]. Welman WG. The genus Solanum (Solanaceae) in southern Africa: subgenus Leptostemonum, the introduced sections Acanthophora and Torva. Bothalia. 2003; 33: 1–18.
. Laemmli UK. ‘Cleavage of structural proteins during the assembly of the head of bacteriophage’, Nature.1970; 227:680.
. Shilpa Satheesh L. ‘Biochemical charecterisation of protease inhibitors from Coccinia indicia W. & A’. 2011; 92-94.
. Mostafa M, Hemayet Hossain, Anwar Hossain M, Pizush KB, Zahurul Haque M. ‘Insecticidal activity of plant extracts against Tribolium castaneum Herbst’. Journal of Advanced Scientific Research. 2012; 3(3): 80-84.
. Pugazhvendan SR, Elumalai PR, Ross, Soundarajan M. ‘Repellent activity of chosen plant species against Tribolium castaneum’. World Journal of Zoology. 2009; 4:188-190.
. Rani AT, Bhanu KR, Chakravarthy AK. ‘Behavioral Responses of Banana Pseudo stem Weevil, Odoiporus longicollis Ilivier (Coleoptera: Curculionidae) to Insect and Plant Volatiles’, Environment & Ecology. 2017; 35 (1):106—112.
. Tinzaara W, Gold CS, Dicke M, Van Huis A, Ragama PE. ‘Host plant odours enhance the responses of adult banana weevil to the synthetic aggregation pheromone Cosmolure’. International Journal of Pest Management. 2007; 53:127—137.
. Prasuna AL, Jyothi KN, Prasad AR, Yadav JL, Padmanaban B. ‘Olfactory responses of banana pseudo stem weevil, Odoiporus longicollis Olivier (Coleoptera: Curculionidae) to semiochemicals from conspecifics and host plant’ Current Science. 2008; 94(7): 896-902.
. Ravi G, Palaniswami MS. ‘Evidence for a female produced sex pheromone in the banana pseudostem weevil, Odoiporus longicollis Olivier’. Current Science. 2002; 83 (7): 893 - 898.
. Holscher D, Dhakshinamoorthy S, Alexandrov T, Becker M, Bretschneider T, Buerkert A, Crecelius AC, Waele D, Elsen A, Heckel DG. ‘Phenalenone-type phytoalexins mediate resistance of banana plants (Musa spp.) to the burrowing nematode Radopholus similis’. Proceedings of the National Academy of Sciences of USA. 2014; 111:105–110.
. Hidalgo W, Chandran JN, Menezes RC, Otálvaro F, Schneider B. ‘Phenylphenalenones protect banana plants from infection by Mycosphaerella fijiensis and are deactivated by metabolic conversion’. Plant, Cell & Environment. 2016; 39: 492–513.
. Holscher WD, Buerkert A Schneide B. ‘Phenylphenalenones accumulate in plant tissues of two Banana cultivars in response to herbivory by the Banana stem weevil’. Plants. 2016; 34: 1-6
. Sahayaraj P, Kombiah P, Dikshit AK, Rathi JM. ‘Chemical constituents of the essential oils of Tephrosia purpurea and Ipomoea carnea and their repellent activity against Odoiporus longicollis’. Journal of the Serbian Chemical Society. 2015; 80 (4): 465–473.
. Pavela R. ‘Insecticidal activity of certain medicinal plants’. Fitoterapia. 2004; 17: 745-749.
. Sahayaraj K, Kombiah P. ‘Olfactory responses of the banana weevil, Odoiporus longicollis (Olivier) (Coleoptera: Curculionidae) against pseudostem and its crude extract’, Journal of Biopesticides. 2009; 2(2): 173–176.
. Gailce LJ, Leelamath M, Nirmal Johson SB. ‘Bionomics and mansagement of the pseudostem weevil Odoiporus longicollis Oliver (Coleoptera: Curculionidae) in banana– A Review’. Agriculture Review. 2008; 29: 185-192.
. Tinzaara W, Tushemereirwe WK, Kashaija I. ‘Efficiency of pheromones and trap types in the capture of the banana weevil Cosmopolites sordidus Germar in Uganda’. Uganda Journal of Agricultural Sciences. 2000; 5:91-97.
. Palanichamy S, Padmanaban B, Mohamed MIF, Mustaffa MM. Microwave Oven Assisted Extraction of Banana pseudostem kairomones as attractant of Odoiporus longicollis Olivier (Coleoptera: Curculionidae): Electroantennogram investigations. Archives in Applied Science and Research. 2011; 3: 213-216.
. Karmegam N, Sakthivadivel M, Anuradha R, Daniel T. ‘Indigenous-plant extracts as larvicidal against Culex quinqefasciatus say’. Bioresource Technology. 1997; 59: 137-140.
. Murugan K, Babu R. ‘Impact of certain plant products and Bacillus thurengiensis Berliner sub sp. Kurstaki on the growth and feeding physiology of Helicoverpa armigera (Hubner)’. Journal of Scientific and Industrial Research. 1998; 57: 757-765.
