87 kDa tomato cystatin exhibits properties of a defense protein and forms protein crystals in prosystemin overexpressing transgenic plants |
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| Institution: | 1. Laboratório de Biotecnologia, Universidade Estadual do Norte Fluminense, 28015-620, Campos dos Goytacazes, Rio de Janeiro, Brazil;2. Laboratório de Química e Função de Proteínas e Peptídeos, Universidade Estadual do Norte Fluminense, 28015-620, Campos dos Goytacazes, Rio de Janeiro, Brazil;3. Laboratório de Biologia Celular e Tecidual, Universidade Estadual do Norte Fluminense, 28015-620, Campos dos Goytacazes, Rio de Janeiro, Brazil;4. Laboratório de Bioquímica e Fisiologia de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, 28015-620, Campos dos Goytacazes, Rio de Janeiro, Brazil;5. Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340, USA;1. Institut de Recherche pour le Développement, UMR Diversité Adaptation et Développement des Plantes (DIADE), 911 Avenue Agropolis, 34394 Montpellier cedex 5, France;2. Laboratoire Mixte International Adaptation des Plantes et Microorganismes Associés aux Stress Environnementaux (LAPSE), Centre de Recherche de Bel Air, BP 1386, Dakar, Senegal;3. Centre d’Etudes Régional pour l’Amélioration de l’Adaptation à la Sècheresse (CERAAS), Institut Sénégalais de Recherches Agricoles, BP 3320, Route de Khombole, Thiès, Senegal;4. Laboratoire Commun de Microbiologie IRD/ISRA/UCAD, Centre de Recherche de Bel Air, BP 1386, Dakar, Senegal;1. Department of Medical Physiology, Faculty of Medicine, Universitas Indonesia, Indonesia;2. Graduate Student of Master Program in Biomedical Science, Faculty of Medicine, Universitas Indonesia, Indonesia;3. Metabolic, Cardiovascular and Aging Research Cluster, The Indonesian Medical Education and Research Institute, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia;4. Division of Endocrinology, Metabolism and Diabetes, Department of Internal Medicine, dr. Cipto Mangunkusumo National General Hospital/Faculty of Medicine Universitas Indonesia, Indonesia;3. From the Division of Cardiovascular Medicine, Department of Medicine, and;5. the Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida 32608 and;4. the Interdisciplinary Institute of the Indian System of Medicine, SRM University, Kattankulathur, Tamil Nadu 603203, India;1. Institute of Plant Sciences, A.R.O., The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel;2. The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel;3. Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, New York, USA;1. Deakin University, Geelong Waurn Ponds Campus, School of Life and Environmental Sciences, Victoria, 3216, Australia;2. Department of Plant Pathology, Faculty of Agriculture, Sher-e-Bangla Agricultural University, Sher-e-Bangla Nagar, Dhaka-1207, Bangladesh;3. Department of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania “Luigi Vanvitelli”, I-81100, Caserta, Italy;4. Department of Chemical Sciences, University of Naples Federico II, I-80126, Naples, Italy;5. Institute for Biochemistry, Biotechnology and Bioinformatics, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany;6. Section Pathology of Woody Plants, Technische Universität München Hans-Carl-von-Carlowitz-Platz 2, 85354, Freising, Germany;1. Department of Food Science, South China University of Technology, Guangzhou 510640, China;2. Department of Food Science, North Carolina State University, Raleigh, NC 27695-7624, United States |
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| Abstract: | Transgenic tomato plants (Lycopersicon esculentum) overexpressing the prosystemin transgene have been shown previously to accumulate a soluble 87 kDa cystatin constitutively. We report here that this protein can be found in a crystalline form which can be purified using a glycerol/sucrose gradient. Midgut homogenate of third-instar larvae of two coleopteran pest insects, Callosobruchus maculatus and Zabrotes subfasciatus, had their proteolytic activity content significantly inhibited by tomato cystatin (TC). In leaves of wild-type tomato plants, cystatin mRNA accumulated systemically in response to wounding, treatment with methyl jasmonate (MJ) and when supplied with systemin, corroborating the anti-herbivorous activity. Accumulation of cystatin mRNA occurred when plants were supplied with chitosan and oligogalacturonic acid fragments (OGA), suggesting an effect of TC against pathogens. Moreover, this protein reduced the growth of two fungi, Fusarium solani and Trichoderma viride in vitro. Taken together, the data reinforce a role for TC in defense response against pests or pathogens. |
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