Proteinase Inhibitors of Pigeonpea cv. BSMR 736: Characterization and Bioefficacy Against Helicoverpa Armigera

Keywords: Proteinase inhibitors, pigeonpea, PPTI, helicoverpa armigera, HaGPs

Abstract

Pigeonpea is an agriculturally important leguminous crop with high vulnerability to insect pest attack specifically, Helicoverpa armigera. The proteinase inhibitors (PIs) mediated host plant resistance against insect pests is a promising sustainable agricultural research practice. The current study was carried out to perceive biochemical characterization of proteinase inhibitors named PPTI in the pigeonpea (cv. BSMR 736). The purification of PPTI from crude protein seed extract was achieved by acetone precipitation, N-LP-IEF, and trypsin affinity chromatography. It was found to inhibit bovine trypsin and HaGPs in vitro. The optimal conditions for inhibition were pH 8 and temperature 40ºC. The PPTI showed four isoinhibitors bands on native, non-reducing and reducing SDS-PAGE in the range of 26.7–19.3 KDa. Upon resolution on two-dimensional gel electrophoresis (2-DE), PPTI produced nine pI variant spots having isoelectric point (pI) 6.6, 6.6, 6.3, 6.1, 5.9, 5.8, 5.7, 5.6 and 5.6. An artificial diet containing PPTI reduced the H. armigera larval weight about 69%, with 25% mortality. For eco-friendly sustainable agricultural practices, natural compounds like PPTI could be expressed in transgenic crops to prevent the invasion of H. armigera in pigeonpea.

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References

‎P.R. Shewry, Tuber storage proteins. Annals of Botany, vol. 91, pp. 755-769, 2003. ‎

‎M. Clemente, M.G. Corigliano, S.A. Pariani, E.F. Sánchez-López, V.A. Sander, V.A. Ramos-‎Duarte, Plant Serine Protease Inhibitors: Biotechnology Application in Agriculture and Molecular ‎Farming. International Journal of Molecular Sciences, vol. 20, no. 6, pp. 1345. ‎

‎M. V. Padul, R. D. Tak, and M. S. Kachole, Protease inhibitor (PI) mediated defense in leaves ‎and flowers of pigeonpea (protease inhibitor mediated defense in pigeonpea). Plant Physiology ‎and Biochemistry, vol. 52, pp. 77–82, 2012. ‎

‎F. D. Leo, Plant-PIS: A database for plant protease inhibitors and their genes. Nucleic Acids ‎Research, vol. 30, no. 1, pp. 347–348, 2002. ‎

‎G. C. Mello, I. A. Desouza, S. Marangoni, J. C. Novello, E. Antunes, and M. L. Macedo, ‎OEDEMATOGENIC activity induced by Kunitz-type inhibitors from Dimorphandra mollis ‎seeds. Toxicon, vol. 47, no. 2, pp. 150–155, 2006. ‎

‎P. Tiffin and B. S. Gaut, Molecular evolution of the wound-induced serine protease inhibitor ‎WIP1 in Zea and related genera. Molecular Biology and Evolution, vol. 18, no. 11, pp. 2092–‎‎2101, 2001. ‎

‎R.-F. QI, Z.-W. SONG, and C.-W. CHI, Structural features and molecular evolution of ‎Bowman-Birk protease inhibitors and their potential application. Acta Biochimica et Biophysica ‎Sinica, vol. 37, no. 5, pp. 283–292, 2005. ‎

‎W. Reed, S.S. Lateef, Pigeonpea: Pest management in the pigeonpea (Nene YL, Hall SD and ‎Sheila VK eds). Oxon, Wallingford, UK: CAB International. pp. 349-374. 1990.‎

‎J.W. Lammers, A. MacLeod, Report of a Pest Risk Analysis Helicoverpa armigera (Hübner, ‎‎1808). 2010-11-17. ‎

‎Sharma, H.C, Cotton Bollworm/Legume Pod Borer, Helicoverpa armigera (Hubner) (Noctuidae: ‎Lepidoptera): Biology and Management. Crop Protection Compendium, Commonwealth ‎Agricultural Bureau International Oxon, UK. pp. 72. 2001.‎

‎K. R. Kranthi, D. R. Jadhav, S. Kranthi, R. R. Wanjari, S. S. Ali, and D. A. Russell, Insecticide ‎resistance in five major insect pests of cotton in India. Crop Protection, vol. 21, no. 6, pp. 449–‎‎460, 2002. ‎

‎H. C. Sharma, G. Pampapathy, S. K. Lanka, and T. J. Ridsdill-Smith, ANTIBIOSIS mechanism ‎of resistance to pod borer, Helicoverpa armigera in wild relatives of chickpea. Euphytica, vol. ‎‎152, no. 1, pp. 135–135, 2006. ‎

‎O. L. Franco, R. C. dos Santos, J. A. N. Batista, A. C. Mendes, M. A. de Araújo, R. G. ‎Monnerat, M. F. Grossi-de-Sá, and S. M. de Freitas, Effects of black-eyed Pea ‎Trypsin/chymotrypsin inhibitor on proteolytic activity and on development of Anthonomus ‎grandis. Phytochemistry, vol. 63, no. 3, pp. 343–349, 2003. ‎

‎C. L. Araújo, I. W. Bezerra, A. S. Oliveira, F. T. Moura, L. L. Macedo, C. E. Gomes, A. E. ‎Barbosa, F. P. Macedo, T. M. Souza, O. L. Franco, C. Bloch-J, and M. P. Sales, In vivo ‎bioinsecticidal activity toward ceratitis capitata (fruit fly) and callosobruchus maculatus (cowpea ‎weevil) and in vitro bioinsecticidal activity toward different orders of insect pests of a trypsin ‎inhibitor purified from tamarind tree (tamarindus indica) seeds. Journal of Agricultural and Food ‎Chemistry, vol. 53, no. 11, pp. 4381–4387, 2005. ‎

M. C. Falco and M. C. Silva-Filho, Expression of soybean proteinase inhibitors in transgenic ‎sugarcane plants: Effects on natural defense against Diatraea saccharalis. Plant ‎Physiology and Biochemistry, vol. 41, no. 8, pp. 761–766, 2003. ‎

‎Y.-M. Kim, Y.-K. Jeong, M.-H. Wang, W.-Y. Lee, and H.-I. Rhee, Inhibitory effect of pine ‎extract on α-glucosidase activity and postprandial hyperglycemia. Nutrition, vol. 21, no. 6, pp. ‎‎756–761, 2005. ‎

‎R. D. Tak, M. V. Padul, and M. S. Kachole, Analysis of sequential accumulation of individual ‎pigeonpea protease inhibitors during seed development. Acta Physiologiae Plantarum, vol. 35, ‎no. 12, pp. 3303–3314, 2013. ‎

‎M. Swathi, V. Lokya, V. Swaroop, N. Mallikarjuna, M. Kannan, A. Dutta-Gupta, and K. ‎Padmasree, Structural and functional characterization of proteinase inhibitors from seeds of ‎Cajanus cajan (cv. ICP 7118), Plant Physiology and Biochemistry, vol. 83, pp. 77–87, 2014.‎ ‎ ‎

‎N. P. Chougule, V. K. Hivrale, P. J. Chhabda, A. P. Giri, and M. S. Kachole, Differential ‎inhibition of Helicoverpa armigera gut proteinases by proteinase inhibitors of pigeonpea ‎‎(Cajanus cajan) and its wild relatives. Phytochemistry, vol. 64, no. 3, pp. 681–687, 2003. ‎

‎M. A. Jongsma, P. L. Bakker, J. Peters, D. Bosch, and W. J. Stiekema, Adaptation of spodoptera ‎exigua larvae to plant proteinase inhibitors by induction of gut proteinase activity insensitive to ‎inhibition. Proceedings of the National Academy of Sciences USA, vol. 92, no. 17, pp. 8041–‎‎8045, 1995. ‎

‎A. G. Patankar, A. P. Giri, A. M. Harsulkar, M. N. Sainani, V. V. Deshpande, P. K. Ranjekar, ‎and V. S. Gupta, Complexity in specificities and expression of Helicoverpa armigera gut ‎proteinases explains polyphagous nature of The insect pest. Insect Biochemistry and Molecular ‎Biology, vol. 31, no. 4-5, pp. 453–464, 2001.

‎S. K. Haq and R. H. Khan, Characterization of a proteinase inhibitor fromcajanus cajan (l.). ‎Journal of Protein Chemistry, vol. 22, no. 6, pp. 543–554, 2003. ‎

‎A. P. Giri and M. S. Kachole, Amylase inhibitors of pigeonpea (Cajanus cajan) seeds. ‎Phytochemistry, vol. 47, no. 2, pp. 197–202, 1998. ‎

‎M. M. Pichare and M. S. Kachole, Detection of electrophoretically separated protease inhibitors ‎using X-ray film. Journal of Biochemical and Biophysical Methods, vol. 28, no. 3, pp. 215–224, ‎‎1994. ‎

‎B. J. Davis, Disc electrophoresis - II method and application to human serum proteins. Annals of ‎the New York Academy of Sciences, vol. 121, no. 2, pp. 404–427, 2006. ‎

‎U. K. LAEMMLI, Cleavage of structural proteins during the Assembly of the head of ‎bacteriophage T4, Nature, vol. 227, no. 5259, pp. 680–685, 1970. ‎

‎M.L. Kakade, N. Simons, I.E. Liener, An evaluation of natural vs. synthetic substrates for ‎measuring the antitryptic activity of soybean samples. Cereal Chemistry, vol. 46, pp. 518-521. ‎‎1969. ‎

‎O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall, Protein measurement with the ‎Folin phenol reagent. Journal of Biological Chemistry, vol. 193, no. 1, pp. 265–275, 1951. ‎

‎P. Christou, T. Capell, A. Kohli, J. A. Gatehouse, and A. M. R. Gatehouse, Recent developments ‎and future prospects in insect pest control in transgenic crops. Trends in Plant Sciences, vol. 11, ‎no. 6, pp. 302–308, 2006. ‎

‎V. V. Mosolov and T. A. Valueva, Proteinase inhibitors in plant biotechnology: A Review. ‎Applied Biochemistry and Microbiology, vol. 44, no. 3, pp. 233–240, 2008. ‎

‎C. L. Nilsson, M. Puchades, A. Westman, K. Blennow, and P. Davidsson, Identification of ‎proteins in a human pleural exudate using two-dimensional preparative liquid-phase ‎electrophoresis and matrix-assisted laser desorption/ionization mass spectrometry. ‎Electrophoresis, vol. 20, pp. 860-865, 1999. ‎

‎T. Q. Shang, J. M. Ginter, M. V. Johnston, B. S. Larsen, and C. N. McEwen, Carrier ampholyte-‎free solution isoelectric focusing as a prefractionation method for the proteomic analysis of ‎complex protein mixtures. Electrophoresis, vol. 24, no. 14, pp. 2359–2368, 2003. ‎

‎E. R. Prasad, H. Merzendorfer, C. Madhurarekha, A. Dutta-Gupta, and K. Padmasree, ‎Bowman−Birk proteinase inhibitor from Cajanus cajan seeds: Purification, characterization, and ‎insecticidal properties. Journal of Agricultural and Food Chemistry, vol. 58, no. 5, pp. 2838–‎‎2847, 2010. ‎

‎R.-X. Li, H. Zhou, S.-J. Li, Q.-H. Sheng, Q.-C. Xia, and R. Zeng, Prefractionation of proteome ‎by liquid isoelectric focusing prior to two-dimensional liquid chromatography mass spectrometric ‎identification. Journal of Proteome Research, vol. 4, no. 4, pp. 1256–1264, 2005. ‎

‎J. N. Losso, The biochemical and functional food properties of the bowman-birk inhibitor. ‎Critical Reviews in Food Science and Nutrition, vol. 48, no. 1, pp. 94–118, 2008. ‎

S. O. O. T. T. A. W. A. T. Benjakul, W. O. N. N. O. P. Visessanguan, and P. A. I. B. O. O. N. ‎Thummaratwasik, Isolation and characterization of trypsin inhibitors from some Thai legume ‎seeds. Journal of Food Biochemistry, vol. 24, no. 2, pp. 107–127, 2000. ‎

S. A. Godbole, T. G. Krishna, and C. R. Bhatia, Purification and characterisation of protease ‎inhibitors from pigeon pea (Cajanus cajan (L) MILLSP) seeds. Journal of the Science of Food ‎and Agriculture, vol. 64, no. 1, pp. 87–93, 1994. ‎

‎Y. N. Sreerama, J. R. Das, D. R. Rao, and L. R. Gowda, Double-headed trypsin/chymotrypsin ‎inhibitors from horse gram (Dolichos Biflorus): Purification, molecular and Kinetic Properties. ‎Journal of Food Biochemistry, vol. 21, no. 1, pp. 461–477, 1997. ‎

‎K. Y. Ee, J. Zhao, A.-ur Rehman, and S. O. Agboola, Purification and characterization of a ‎Kunitz-type trypsin inhibitor from acacia victoriae bentham seeds. Journal of Agricultural and ‎Food Chemistry, vol. 57, no. 15, pp. 7022–7029, 2009. ‎

‎E. Ferrasson, L. Quillien, and J. Gueguen, Proteinase inhibitors from PEA Seeds: purification ‎and characterization. Journal of Agricultural and Food Chemistry, vol. 45, no. 1, pp. 127–131, ‎‎1997. ‎

‎R. Kansal, R. N. Gupta, K. R. Koundal, K. Kuhar, and V. K. Gupta, Purification, ‎characterization and evaluation of insecticidal potential of trypsin inhibitor from mungbean ‎‎(Vigna radiata L. Wilczek) seeds. Acta Physiologiae Plantarum, vol. 30, no. 6, pp. 761–768, ‎‎2008. ‎

‎K. A. Johnston, M. J. Lee, J. A. Gatehouse, and J. H. Anstee, The partial purification and ‎characterisation of serine protease activity in midgut of larval Helicoverpa armigera. Insect ‎Biochemistry, vol. 21, no. 4, pp. 389–397, 1991. ‎

‎M. H. Ling, H. Y. Qi, and C. W. Chi, Protein, cDNA, and genomic DNA sequences of the towel ‎gourd trypsin inhibitor. A squash family inhibitor. Journal of Biological Chemistry, vol. 268, no. ‎‎2, pp. 810–814, 1993. ‎

‎B. Prakash and M. R. Murthy, Source and target enzyme signature in serine protease inhibitor ‎active site sequences. Journal of Biosciences, vol. 22, no. 5, pp. 555–565, 1997. ‎

‎F. K. Shaikh, S. W. Hamad, S. W. Hamad, and A. A. Shinde, In vitro screening of seed extracts ‎of medicinal plants for protease inhibitory activity. Cihan University-Erbil Scientific Journal, vol. ‎‎3, no. 1, pp. 61–65, 2019. ‎

‎A. A. Shinde, F. K. Shaikh, and M. S. Kachole, Exploration of amylases producing competency ‎of Helicoverpa armigera gut bacterial strain, Bacillus subtilis RTSBA6 6.00. Cihan University-‎Erbil Scientific Journal, vol. 3, no. 1, pp. 75–79, 2019. ‎https://doi:10.24086/cuesj.v3n1y2019.pp75-79

Published
2022-08-01
How to Cite
1.
Shaikh F, Bradosty S, Gadge P, Jaiwal B, Padul M, Patil A. Proteinase Inhibitors of Pigeonpea cv. BSMR 736: Characterization and Bioefficacy Against Helicoverpa Armigera. cuesj [Internet]. 1Aug.2022 [cited 28Mar.2024];6(2):9-6. Available from: https://journals.cihanuniversity.edu.iq/index.php/cuesj/article/view/685
Section
Research Article