Preloader

TcTI, a Kunitz-type trypsin inhibitor from cocoa associated with defense against pathogens

  • 1.

    Clemente, M. et al. Plant serine protease inhibitors: Biotechnology application in agriculture and molecular farming. Int. J. Mol. Sci. 20(6), 1345 (2019).

    CAS 
    PubMed Central 

    Google Scholar 

  • 2.

    Lima, A. M. et al. A new Piper nigrum cysteine proteinase inhibitor, PnCPI, with antifungal activity: Molecular cloning, recombinant expression, functional analyses and molecular modeling. Planta 252(2), 16 (2020).

    CAS 
    PubMed 

    Google Scholar 

  • 3.

    Ryan, C. A. Protease inhibitors in plants: Genes for improving defenses against insects and pathogens. Annu. Rev. Phytopathol. 28(1), 425–449 (1990).

    CAS 

    Google Scholar 

  • 4.

    Ma, Y., Zhao, Q., Lu, M.-Z. & Wang, J. Kunitz-type trypsin inhibitor gene family in Arabidopsis and Populus trichocarpa and its expression response to wounding and herbivore in Populus nigra. Tree Genet. Genomes. 7(2), 431–441 (2011).

    Google Scholar 

  • 5.

    Rufino, F. P. S. et al. Inhibitory effects of a Kunitz-type inhibitor from Pithecellobium dumosum [Benth] seeds against insect-pests’ digestive proteinases. Plant Physiol. Biochem. 63, 70–76 (2013).

    CAS 
    PubMed 

    Google Scholar 

  • 6.

    Bateman, K. S. & James, M. N. G. Plant protein proteinase inhibitors: Structure and mechanism of inhibition. Curr. Protein Pept. Sci. 12(5), 341–347 (2011).

    CAS 

    Google Scholar 

  • 7.

    Hartl, M., Giri, A. P., Kaur, H. & Baldwin, I. T. Serine protease inhibitors specifically defend Solanum nigrum against generalist herbivores but do not influence plant growth and development. Plant Cell 22(12), 4158–4175 (2010).

    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 8.

    Bendre, A. D., Ramasamy, S. & Suresh, C. G. Analysis of Kunitz inhibitors from plants for comprehensive structural and functional insights. Int. J. Biol. Macromol. 113, 933–943 (2018).

    CAS 
    PubMed 

    Google Scholar 

  • 9.

    Jamal, F., Pandey, P. K., Singh, D. & Khan, M. Y. Serine protease inhibitors in plants: Nature’s arsenal crafted for insect predators. Phytochem. Rev. 12(1), 1–34 (2013).

    CAS 

    Google Scholar 

  • 10.

    Rawlings, N. D., Barrett, A. J. & Bateman, A. MEROPS: The peptidase database. Nucleic Acids Res. 38(suppl_1), D227–D233 (2010).

    CAS 
    PubMed 

    Google Scholar 

  • 11.

    Ribeiro, J. K. C., Cunha, D. D. S., Fook, J. M. S. L. L. & Sales, M. P. New properties of the soybean trypsin inhibitor: Inhibition of human neutrophil elastase and its effect on acute pulmonary injury. Eur. J. Pharmacol. 644(1–3), 238–244 (2010).

    CAS 
    PubMed 

    Google Scholar 

  • 12.

    Alibeik, S., Zhu, S., Yau, J. W., Weitz, J. I. & Brash, J. L. Surface modification with polyethylene glycol–corn trypsin inhibitor conjugate to inhibit the contact factor pathway on blood-contacting surfaces. Acta Biomater. 7(12), 4177–4186 (2011).

    CAS 
    PubMed 

    Google Scholar 

  • 13.

    Pereira, K. R. B. et al. Passion fruit flowers: Kunitz trypsin inhibitors and cystatin differentially accumulate in developing buds and floral tissues. Phytochemistry 72(16), 1955–1961 (2011).

    CAS 
    PubMed 

    Google Scholar 

  • 14.

    Arnaiz, A. et al. Arabidopsis Kunitz trypsin inhibitors in defense against spider mites. Front Plant Sci. 10(9), 986 (2018).

    Google Scholar 

  • 15.

    Telang, M. et al. Bitter gourd proteinase inhibitors: Potential growth inhibitors of Helicoverpa armigera and Spodoptera litura. Phytochemistry 63(6), 643–652 (2003).

    CAS 
    PubMed 

    Google Scholar 

  • 16.

    Wakasa, Y, & Takaiwa, F. Seed storage proteins, in Brenner’s Encyclopedia of Genetics. 346–348 (Elsevier, 2013) https://linkinghub.elsevier.com/retrieve/pii/B9780123749840013784

  • 17.

    Jongsma, M. A. & Bolter, C. The adaptation of insects to plant protease inhibitors. J. Insect. Physiol. 43(10), 885–895 (1997).

    CAS 
    PubMed 

    Google Scholar 

  • 18.

    Leo, F. D. PLANT-PIs: A database for plant protease inhibitors and their genes. Nucleic Acids Res. 30(1), 347–348 (2002).

    ADS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 19.

    Zavala, J. A., Patankar, A. G., Gase, K., Hui, D. & Baldwin, I. T. Manipulation of endogenous trypsin proteinase inhibitor production in Nicotiana attenuata demonstrates their function as antiherbivore defenses. Plant Physiol. 134(3), 1181–1190 (2004).

    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 20.

    Sivakumar, S. et al. Screening and purification of a novel trypsin inhibitor from prosopis juliflora seeds with activity toward pest digestive enzymes. Protein Pept. Lett. 12(6), 561–565 (2005).

    CAS 
    PubMed 

    Google Scholar 

  • 21.

    Haq, S. K., Atif, S. M. & Khan, R. H. Protein proteinase inhibitor genes in combat against insects, pests, and pathogens: Natural and engineered phytoprotection. Arch. Biochem. Biophys. 431(1), 145–159 (2004).

    CAS 
    PubMed 

    Google Scholar 

  • 22.

    Huang, H. et al. NtKTI1, a Kunitz trypsin inhibitor with antifungal activity from Nicotiana tabacum, plays an important role in tobacco’s defense response: NtKTI1 participates in tobacco’s fungal defense. FEBS J. 277(19), 4076–4088 (2010).

    CAS 
    PubMed 

    Google Scholar 

  • 23.

    Ruan, J.-J., Chen, H., Shao, J.-R., Wu, Q. & Han, X.-Y. An antifungal peptide from Fagopyrum tataricum seeds. Peptides 32(6), 1151–1158 (2011).

    CAS 
    PubMed 

    Google Scholar 

  • 24.

    Dunaevsky, Y. E. et al. The anionic protease inhibitor BWI-1 from buckwheat seeds. Kinetic properties and possible biological role. Physiol. Plant. 101(3), 483–488 (1997).

    CAS 

    Google Scholar 

  • 25.

    Ruan, J. et al. Molecular cloning and structure-function analysis of a trypsin inhibitor from tartary buckwheat and its application in combating phytopathogenic fungi. Agronomy 8(4), 46 (2018).

    Google Scholar 

  • 26.

    Revina, T. A., Speranskaya, A. S., Kladnitskaya, G. V., Shevelev, A. B. & Valueva, T. A. Subtilisin protein inhibitor from potato tubers. Biochem. Mosc. 69(10), 1092–1098 (2004).

    CAS 

    Google Scholar 

  • 27.

    Revina, T. A., Gerasimova, N. G., Kladnitskaya, G. V., Chalenko, G. I. & Valueva, T. A. Effect of proteinaceous proteinase inhibitors from potato tubers on the growth and development of phytopathogenic microorganisms. Appl. Biochem. Microbiol. 44(1), 89–92 (2008).

    CAS 

    Google Scholar 

  • 28.

    Melo, F. R. et al. Inhibition of trypsin by cowpea thionin: Characterization, molecular modeling, and docking. Prot. Struct. Funct. Bioinf. 48(2), 311–319 (2002).

    CAS 

    Google Scholar 

  • 29.

    Gesteira, A. S. et al. Comparative analysis of expressed genes from cacao meristems infected by moniliophthora perniciosa. Ann. Bot. 100(1), 129–140 (2007).

    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 30.

    Sousa Silva, C. R. & Figueira, A. Phylogenetic analysis of Theobroma [Sterculiaceae] based on Kunitz-like trypsin inhibitor sequences. Plant Syst. Evol. 250(1–2), 93–104 (2005).

    CAS 

    Google Scholar 

  • 31.

    Paulillo, L. C. M. S. et al. Evaluation of in vitro and in vivo effects of semipurified proteinase inhibitors from theobroma seeds on midgut protease activity of lepidopteran pest insects: Evaluation of theobroma proteinase inhibitors. Arch. Insect. Biochem. Physiol. 81(1), 34–52 (2012).

    CAS 
    PubMed 

    Google Scholar 

  • 32.

    Santana, L. S. et al. TcCYS4, a cystatin from cocoa, reduces necrosis triggered by MpNEP2 in tobacco plants. Genet. Mol. Res. 13(3), 7636–7648 (2014).

    CAS 
    PubMed 

    Google Scholar 

  • 33.

    Pirovani, C. P. et al. Theobroma cacao cystatins impair Moniliophthora perniciosa mycelial growth and are involved in postponing cell death symptoms. Planta 232(6), 1485–1497 (2010).

    CAS 
    PubMed 

    Google Scholar 

  • 34.

    Petersen, T. N., Brunak, S., von Heijne, G. & Nielsen, H. SignalP 4.0: Discriminating signal peptides from transmembrane regions. Nat. Meth. 8(10), 785–786 (2011).

    CAS 

    Google Scholar 

  • 35.

    Finn, R. D. et al. The Pfam protein families database: Towards a more sustainable future. Nucleic Acids Res. 44(D1), D279–D285 (2016).

    CAS 
    PubMed 

    Google Scholar 

  • 36.

    Patro, R., Duggal, G., Love, M. I., Irizarry, R. A. & Kingsford, C. Salmon provides fast and bias-aware quantification of transcript expression. Nat. Methods. 14(4), 417–419 (2017).

    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 37.

    Teixeira, P. J. P. L. et al. High-resolution transcript profiling of the atypical biotrophic interaction between Theobroma cacao and the fungal pathogen Moniliophthora perniciosa. Plant Cell 26(11), 4245–4269 (2014).

    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 38.

    Team RC. R: A language and environment for statistical computing (2013).

  • 39.

    Argout, X. et al. Towards the understanding of the cocoa transcriptome: Production and analysis of an exhaustive dataset of ESTs of Theobroma cacao L. generated from various tissues and under various conditions. BMC Genomics 9(1), 512 (2008).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 40.

    Sambrook, J., Fritsch, E. F. & Maniatis, T. Molecular Cloning: A Laboratory Manual (Cold spring harbor laboratory press, 1989).

    Google Scholar 

  • 41.

    Louis-Jeune, C., Andrade-Navarro, M. A. & Perez-Iratxeta, C. Prediction of protein secondary structure from circular dichroism using theoretically derived spectra. Prot. Struct. Funct. Bioinf. 80(2), 374–381 (2012).

    CAS 

    Google Scholar 

  • 42.

    Guex, N. & Peitsch, M. C. SWISS-MODEL and the swiss-Pdb viewer: An environment for comparative protein modeling. Electrophoresis 18(15), 2714–2723 (1997).

    CAS 
    PubMed 

    Google Scholar 

  • 43.

    Gahloth, D., Selvakumar, P., Shee, C., Kumar, P. & Sharma, A. K. Cloning, sequence analysis and crystal structure determination of a miraculin-like protein from Murraya koenigii. Arch. Biochem. Biophys. 494(1), 15–22 (2010).

    CAS 
    PubMed 

    Google Scholar 

  • 44.

    Laskowski, R., Macarthur, M. W., Moss, D. S. & Thornton, J. PROCHECK: A program to check the stereochemical quality of protein structures. J. Appl. Crystallogr. 26, 283–291 (1993).

    CAS 

    Google Scholar 

  • 45.

    Surujdeo-Maharaj, S., Umaharan, P., Butler, D. R. & Sreenivasan, T. N. An optimized screening method for identifying levels of resistance to Crinipellis perniciosa in cocoa [Theobroma cacao]. Plant Pathol. 52(4), 464–475 (2003).

    Google Scholar 

  • 46.

    Pirovani, C. P. et al. Protein extraction for proteome analysis from cacao leaves and meristems, organs infected byMoniliophthora perniciosa, the causal agent of the witches’ broom disease. Electrophoresis 29(11), 2391–2401 (2008).

    CAS 
    PubMed 

    Google Scholar 

  • 47.

    dos Santos, E. C., Pirovani, C. P., Correa, S. C., Micheli, F. & Gramacho, K. P. The pathogen Moniliophthora perniciosa promotes differential proteomic modulation of cacao genotypes with contrasting resistance to witches´ broom disease. BMC Plant Biol. 20(1), 1 (2020).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 48.

    Vilela, M., Mendes, S. M., Valicente, F. H., de Carvalho, S. S., dos Santos, A. E., dos Santos, C. A., et al. Metodologia para criação e manutenção de Helicoverpa armigera em laboratório. Embrapa Milho E Sorgo-Circ Téc INFOTECA-E (2014).

  • 49.

    Halder, J., Srivastava, C., Dhingra, S. & Dureja, P. Effect of Essential Oils on Feeding, Survival, Growth and Development of Third Instar Larvae of Helicoverpa armigera Hubner. Natl. Acad. Sci Lett. 35, 271–276 (2012).

    Google Scholar 

  • 50.

    Caldas, M. M. & Perz, S. Agro-terrorism? The causes and consequences of the appearance of witch’s broom disease in cocoa plantations of southern Bahia, Brazil. Geoforum 47, 147–157 (2013).

    Google Scholar 

  • 51.

    Oliva, M. L. V., Silva, M. C. C., Sallai, R. C., Brito, M. V. & Sampaio, M. U. A novel subclassification for Kunitz proteinase inhibitors from leguminous seeds. Biochimie 92(11), 1667–1673 (2010).

    CAS 
    PubMed 

    Google Scholar 

  • 52.

    Argout, X. et al. The genome of Theobroma cacao. Nat. Genet. 43(2), 101–108 (2011).

    CAS 
    PubMed 

    Google Scholar 

  • 53.

    Senthilkumar, R. & Yeh, K.-W. Multiple biological functions of sporamin related to stress tolerance in sweet potato [Ipomoea batatas Lam]. Biotechnol. Adv. 30(6), 1309–1317 (2012).

    CAS 
    PubMed 

    Google Scholar 

  • 54.

    Effendy, J., LaBonte, D. R. & Efendi, D. Isolation and characterization of cDNA clones encoding a novel subfamily sporamin B in sweet potato. Biodiversitas J. Biol. Divers. 20(10), 1–9 (2019).

    Google Scholar 

  • 55.

    Cardoso, T. H. S. et al. TcCYPR04, a cacao papain-like cysteine-protease detected in senescent and necrotic tissues interacts with a cystatin TcCYS4. Permyakov EA, editor. PLoS ONE 10(12), e0144440 (2015).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 56.

    Freitas, A. C. O. et al. The activity of TcCYS4 modified by variations in pH and temperature can affect symptoms of witches’ broom disease of cocoa, caused by the fungus moniliophthora perniciosa. Margis R, editor. PLoS ONE 10(4), e0121519 (2015).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 57.

    Chen, Z., Cao, Z., Li, W. & Wu, Y. Cloning and characterization of a novel Kunitz-type inhibitor from scorpion with unique cysteine framework. Toxicon 72, 5–10 (2013).

    CAS 
    PubMed 

    Google Scholar 

  • 58.

    Deshimaru, M., Hanamoto, R., Kusano, C., Yoshimi, S. & Terada, S. Purification and characterization of proteinase inhibitors from wild soja [Glycine soja ] seeds. Biosci. Biotechnol. Biochem. 66(9), 1897–1903 (2002).

    CAS 
    PubMed 

    Google Scholar 

  • 59.

    Oddepally, R., Sriram, G. & Guruprasad, L. Purification and characterization of a stable Kunitz trypsin inhibitor from Trigonella foenum-graecum [fenugreek] seeds. Phytochemistry 96, 26–36 (2013).

    CAS 
    PubMed 

    Google Scholar 

  • 60.

    de Oliveira, C. F. R. et al. Purification and biochemical properties of a Kunitz-type trypsin inhibitor from Entada acaciifolia [Benth.] seeds. Process Biochem. 47(6), 929–935 (2012).

    Google Scholar 

  • 61.

    Orona-Tamayo, D. et al. Exclusive rewards in mutualisms: Ant proteases and plant protease inhibitors create a lock-key system to protect Acacia food bodies from exploitation. Mol. Ecol. 22(15), 4087–4100 (2013).

    CAS 
    PubMed 

    Google Scholar 

  • 62.

    Klomklao, S., Benjakul, S., Kishimura, H. & Chaijan, M. Extraction, purification and properties of trypsin inhibitor from Thai mung bean [Vigna radiata L.] R. Wilczek]. Food Chem. 129(4), 1348–1354 (2011).

    CAS 

    Google Scholar 

  • 63.

    Brandão-Costa, R. M. P., Araújo, V. F. & Porto, A. L. F. CgTI, a novel thermostable Kunitz trypsin-inhibitor purified from Cassia grandis seeds: Purification, characterization and termiticidal activity. Int. J. Biol. Macromol. 118, 2296–2306 (2018).

    PubMed 

    Google Scholar 

  • 64.

    Bhattacharyya, A. & Babu, C. R. Purification and biochemical characterization of a serine proteinase inhibitor from Derris trifoliata Lour. seeds: Insight into structural and antimalarial features. Phytochemistry 70(6), 703–712 (2009).

    CAS 
    PubMed 

    Google Scholar 

  • 65.

    Shamsi, T. N. et al. Allium sativum protease inhibitor: A novel kunitz trypsin inhibitor from garlic is a new comrade of the serpin family. PLoS ONE 11(11), e0165572 (2016).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 66.

    Liao, H. et al. A trypsin inhibitor from Cassia obtusifolia seeds: Isolation, characterization and activity against Pieris rapae. Biotechnol. Lett. 29(4), 653–658 (2007).

    CAS 
    PubMed 

    Google Scholar 

  • 67.

    Chaudhary, N. S. et al. Purification and characterization of a trypsin inhibitor from Putranjiva roxburghii seeds. Phytochemistry 69(11), 2120–2126 (2008).

    CAS 
    PubMed 

    Google Scholar 

  • 68.

    Lopes, J. L. S. et al. Physico-chemical and antifungal properties of protease inhibitors from Acacia plumosa. Phytochemistry 70(7), 871–879 (2009).

    CAS 
    PubMed 

    Google Scholar 

  • 69.

    Meester, P. D., Brick, P., Lloyd, L. F., Blow, D. M. & Onesti, S. Structure of the Kunitz-type soybean trypsin inhibitor [STI]: Implication for the interactions between members of the STI family and tissue-plasminogen activator. Acta Crystallogr. D Biol. Crystallogr. 54(4), 589–597 (1998).

    PubMed 

    Google Scholar 

  • 70.

    Zhou, D. et al. Crystal structures of a plant trypsin inhibitor from Enterolobium contortisiliquum [EcTI] and of Its complex with bovine trypsin. Kobe B, editor. PLoS ONE 8(4), e62252 (2013).

    ADS 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 71.

    Patthy, A., Molnár, T., Porrogi, P., Naudé, R. & Gráf, L. Isolation and characterization of a protease inhibitor from Acacia karroo with a common combining loop and overlapping binding sites for chymotrypsin and trypsin. Arch. Biochem. Biophys. 565, 9–16 (2015).

    CAS 
    PubMed 

    Google Scholar 

  • 72.

    Azarkan, M., Dibiani, R., Goormaghtigh, E., Raussens, V. & Baeyens-Volant, D. The papaya Kunitz-type trypsin inhibitor is a highly stable β-sheet glycoprotein. Biochim. Biophys. Acta BBA Prot. Proteomics. 1764(6), 1063–1072 (2006).

    CAS 

    Google Scholar 

  • 73.

    da Ramos, V. S. et al. Molecular cloning and insecticidal effect of Inga laurina trypsin inhibitor on Diatraea saccharalis and Heliothis virescens. Comp. Biochem. Physiol. Part C Toxicol. Pharmacol. 156(3–4), 148–158 (2012).

    CAS 

    Google Scholar 

  • 74.

    Philippe, R. N., Ralph, S. G., Külheim, C., Jancsik, S. I. & Bohlmann, J. Poplar defense against insects: Genome analysis, full-length cDNA cloning, and transcriptome and protein analysis of the poplar Kunitz-type protease inhibitor family. New Phytol. 184(4), 865–884 (2009).

    CAS 
    PubMed 

    Google Scholar 

  • 75.

    Foster, A. J., Pelletier, G., Tanguay, P. & Séguin, A. Transcriptome analysis of poplar during leaf spot infection with sphaerulina spp. Stromvik M, editor. PLoS ONE 10(9), e0138162 (2015).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 76.

    Guerra, D. et al. Post-transcriptional and post-translational regulations of drought and heat response in plants: A spider’s web of mechanisms. Front Plant Sci. https://doi.org/10.3389/fpls.2015.00057/abstract (2015).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • 77.

    Dias, C. V. et al. Hydrogen peroxide formation in cacao tissues infected by the hemibiotrophic fungus Moniliophthora perniciosa. Plant Physiol. Biochem. 49(8), 917–922 (2011).

    CAS 
    PubMed 

    Google Scholar 

  • 78.

    Alves, A. M. M., Reis, S. P. M., Gramacho, K. P. & Micheli, F. The glutathione peroxidase family of Theobroma cacao: Involvement in the oxidative stress during witches’ broom disease. Int. J. Biol. Macromol. 164, 3698–3708 (2020).

    Google Scholar 

  • 79.

    Pereira Menezes, S. et al. The pathogenesis-related protein PR-4b from Theobroma cacao presents RNase activity, Ca2+ and Mg2+ dependent-DNase activity and antifungal action on Moniliophthora perniciosa. BMC Plant Biol. 14(1), 161 (2014).

    PubMed 
    PubMed Central 

    Google Scholar 

  • 80.

    Freire, L. et al. Tc PHYLL, a cacao phylloplanin expressed in young tissues and glandular trichomes. Physiol. Mol. Plant Pathol. 100, 126–135 (2017).

    CAS 

    Google Scholar 

  • 81.

    Santana, J. O. et al. Characterization of the legumains encoded by the genome of Theobroma cacao L. Plant Physiol. Biochem. 98, 162–170 (2016).

    CAS 
    PubMed 

    Google Scholar 

  • 82.

    de Oliveira, C. G. et al. Involvement of calcium oxalate degradation during programmed cell death in Theobroma cacao tissues triggered by the hemibiotrophic fungus Moniliophthora perniciosa. Plant Sci. 173(2), 106–117 (2007).

    Google Scholar 

  • 83.

    Talyzina, N. M. & Ingvarsson, P. K. Molecular evolution of a small gene family of wound inducible Kunitz trypsin inhibitors in populus. J. Mol. Evol. 63(1), 108–119 (2006).

    ADS 
    CAS 
    PubMed 

    Google Scholar 

  • 84.

    Tsukuda, S., Gomi, K., Yamamoto, H. & Akimitsu, K. Characterization of cDNAs encoding two distinct miraculin-like proteins and stress-related modulation of the corresponding mRNAs in citrus jambhiri lush. Plant Mol. Biol. 60(1), 125–136 (2006).

    CAS 
    PubMed 

    Google Scholar 

  • 85.

    Talon, M. & Gmitter, F. G. Citrus genomics. Int. J. Plant Genomics. 2008, 1–17 (2008).

    Google Scholar 

  • 86.

    Cruz, A. C. B. et al. Bioinsecticidal activity of a novel Kunitz trypsin inhibitor from Catanduva [Piptadenia moniliformis] seeds. Plant Physiol. Biochem. 70, 61–68 (2013).

    CAS 
    PubMed 

    Google Scholar 

  • 87.

    Mehmood, S. et al. Model prediction of a Kunitz-type trypsin inhibitor protein from seeds of Acacia nilotica L. with strong antimicrobial and insecticidal activity. Turk. J. Biol. Turk. Biyol. Derg. 44(4), 188–200 (2020).

    CAS 

    Google Scholar 

  • 88.

    Pandey, P. K., Singh, D. & Jamal, F. The influence of a 21 kda kunitz-type trypsin inhibitor from nonhost madras thorn, pithecellobium dulce, seeds on H. armigera [hübner] [lepidoptera: noctuidae]: Influence of Kunitz-Type Trypsin Inhibitor from Manila Tamarind on H. armigera. Arch. Insect Biochem. Physiol. 89(1), 18–34 (2015).

    CAS 
    PubMed 

    Google Scholar 

  • 89.

    Raubenheimer, D. & Simpson, S. J. Nutrient balancing in grasshoppers: Behavioural and physiological correlates of dietary breadth. J. Exp. Biol. 206(10), 1669–1681 (2003).

    CAS 
    PubMed 

    Google Scholar 

  • Source link