c3518cb17d976b8

تأثیر لکتین استخراج‌شده از قارچ Sclerotinia sclerotiorum بر مرگ‌ومیر و پارامترهای فیزیولوژیک لاروهای پروانۀ سفیده بزرگ کلم، Pieris brassicae (Lepidoptera: Pieridae)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه گیاه‌پزشکی، دانشکدۀ علوم کشاورزی، دانشگاه گیلان، رشت

2 استادیار، گروه گیاه‌پزشکی، دانشکدۀ علوم کشاورزی، دانشگاه گیلان، رشت

3 دانشیار، گروه گیاه‌پزشکی، دانشکدۀ علوم کشاورزی، دانشگاه گیلان، رشت

4 استاد گروه زراعت و اصلاح نباتات، دانشکدۀ علوم کشاورزی، دانشگاه گیلان، رشت

5 کارشناس آزمایشگاه، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت

چکیده

در این پژوهش، لکتین استخراج‌شده از قارچ بیمارگر گیاهی Sclerotinia sclerotiorum، به رژیم غذایی لاروهای سن سوم پروانۀ سفیده بزرگ اضافه گردید و پس از 5 روز تأثیر آن بر مرگ‌ومیر، تغذیه و متابولیسم حد واسط آنها بررسی شد. غلظت‌های 5/0، 1 و 2 میلی‌گرم بر میلی‌لیتر لکتین مذکور به مرگ‌ومیر 20 تا 66 درصدی در لاروهای تیمارشده منجر شد. شاخص‌های تغذیۀ لاروهای تیمارشده نسبت به شاهد کاهش معناداری نشان دادند. میزان هزینۀ متابولیک نیز در لاروهای تیمارشده با لکتین نسبت به شاهد افزایش معناداری را نشان داد. فعالیت آنزیم‌های گوارشی همچون آلفا- آمیلاز، گلوکوزیدازها، لیپاز، پروتئازهای عمومی و اختصاصی (سرین و اگزوپپتیداز) لاروهای تغذیه‌شده با غلظت‌های مختلف لکتین نیز کاهش معناداری نسبت به شاهد نشان داد. اگرچه فعالیت آسپارتات آمینو ترانسفراز و گاما- گلوتامیل ترانسفراز در لاروهای تغذیه‌شده روی غلظت‌های 1 و 2 میلی‌گرم بر میلی‌لیتر لکتین افزایش معناداری را نشان داد، فعالیت آلانین آمینوترانسفراز کاهش معناداری نسبت به شاهد داشت. فعالیت اسید فسفاتاز تفاوت معناداری را بین لاروهای شاهد و تیمار نداشت، اما فعالیت آلکالین فسفاتاز کاهش معناداری را نشان داد. فعالیت لاکتات دهیدروژناز در لاروهای تغذیه‌شده روی غلظت 2 میلی‌گرم بر میلی‌لیتر لکتین افزایش معناداری داشت، اما فعالیت آلدولاز در لاروهای شاهد و تغذیه‌شده از غلظت 5/0 میلی‌گرم بر میلی‌لیتر لکتین بیشتر از دیگر تیمار‌ها بود. میزان درشت مولکول‌های ذخیره‌ای، پروتئین، تری‌گلیسرید و گلیکوژن در لاروهای تغذیه‌شده با لکتین کمتر از شاهد بود. نتایج پژوهش حاضر نشان می‌دهد که لکتین قارچ Sclerotinia sclerotiorum به مرگ‌ومیر و اختلال در فیزیولوژی گوارش و متابولیسم حد واسط لاروهای سفیدۀ بزرگ کلم منجر می‌شود و می‌تواند به عنوان یک مولکول سمی در کنترل این آفت به‌کار رود.

کلیدواژه‌ها


عنوان مقاله [English]

Effect of an extracted lectin from Sclerotinia sclerotiorum de Bary on mortality and physiological parameters in the larvae of large cabbage butterfly, Pieris brassicae (Lepidoptera: Pieridae)

نویسندگان [English]

  • Zeynab Alborzi 1
  • Arash Zibaee 2
  • Jalal Jalali Sendi 3
  • Azadeh Karimi Malati 3
  • Babak Rabiee 4
  • Mehdi Salimi 5
1 M. Sc. Student, Department of Plant Protection, Faculty of Agriculture, University of Guilan, Rasht, Iran
2 Assistant Professor, Department of Plant Protection, Faculty of Agriculture, University of Guilan, Rasht, Iran
3 Associate Professor, Department of Plant Protection, Faculty of Agriculture, University of Guilan, Rasht, Iran
4 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Guilan, Rasht, Iran
5 Library Expert
چکیده [English]

In the current study, the extracted lectin from plant pathogenic fungus, Sclerotinia sclerotiorum was added to diet of third larval instars of Large cabbage butterfly, then its effects were determined on mortality, feeding and intermediary metabolism. Concentrations of 0.5, 1 and 2 mg/ml of the lectin caused 20-60% of mortality in the treated larvae. Amounts of nutritional indices in the treated larvae significantly decreased versus control although level of Metabolic cost in the treated larvae showed statistical increase versus control. Activities of the digestive enzymes such as alpha-amylase, glucosidases, lipase, general and specific proteases (Serine and Exopeptidases) decreased in the fed larvae on lectin concentrations versus control. Although activities of aspartate aminotransferase and γ-glutamyl transferase significantly increased in the concentrations of 1 and 2 mg/ml but activity of alanine aminotransferase had a significant decrease versus control. Activity of acid phosphatase had no statistical differences among treatments and control but activity of alkaline phosphatase was significantly lower than control larvae. Activity of lactate dehydrogenase in the fed larvae on the concentration of 2 mg/ml had significant increase but activities of aldolase in the control and fed larvae on 0.5 mg/ml of lectin were higher than those of other treatments. Amounts of storage macromolecules such as protein, triglyceride and glycogen in the fed larvae on the lectin were lower than those of control so that it was more significant in the concentration of 2 mg/ml. Results of the current study demonstrated that the lectin from S. clerotiorumcaused mortality and interference in digestive physiology and intermediary metabolism of P. Brassicae larvae so it could be used as a toxic molecule to control of the pest.

کلیدواژه‌ها [English]

  • Lectin
  • Sclerotinia sclerotiorum
  • Pieris brassicae
  • Digestive physiology
  • Intermediary metabolism
Agrios, G. N. (2005). Plant Pathology. 5th ed.. Burlington, MA: Elsevier Academic Press. 546-550.
Alborzi, Z. & Zibaee, A. (2014). Purification of Sclerotinia sclerotiorum agglutinin via affinity chromatography. Austin Chromatography (IN PRESS).
Bernfeld P. (1955). Amylases, α and β. Methods in Enzymology, 1, 149-158.
Bessey, O.A., Lowry, O.H. & Brock, M.J. (1946). A method for the rapid determination of alkaline phosphatase with five cubic millimeters of serum. Journal of Biological Chemistry, 164, 321-329.
Candy, L., Van Damme, E.J.M., Peumans, W.J., Menu-Bouaouiche, L., Erard, M. & Rougé, P. (2003). Structural and functional characterization of the GalNAc/Galspecific lectin from the phytopathogenic ascomycete Sclerotinia sclerotiorum (Lib.) de Bary. Biochemistry and Biophysics Research Communication, 308, 396-402.
Chang, H. H. & Kozub, G. C. (1994). Germination of immature and mature sclerotia of Sclerotinia sclerotiorum. Phytopathology, 84, 246-250.
Chen, C. M. (2008). Inducible direct plant defense against insect herbivores: A review. Insect Science, 15, 101-114.
Chun, Y. & Yin, Z. D. (1998). Glycogen assay for diagnosis of female genital Chlamydia trachomatis infection. Journal of Clinical Microbiology, 36, 1081-1082.
Coelho M.B., Marangoni S. & Macedo M.L.R. (2007). Insecticidal action of Annona coriacea lectin against the flour moth Anagasta kuehniella and the rice moth Corcyra cephalonica (Lepidoptera: Pyralidae). Comparative Biochemistry and Physiology Part C, 146, 406-414.
de Oliveira C.F.T., Luz L.A., Paiv P.M.G., Coelho L.C.B.B., Marangoni S. & Macedo M.L.R. (2011). Evaluation of seed coagulant Moringa oleifera lectin (cMoL) as a bioinsecticidal tool with potential for the control of insects. Process Biochemistry, 46, 498-504.
Elpidina, E.N., Vinokurov, K.S., Gromenko, V.A., Rudenskaya, Y.A., Dunaevsky, Y.E. & Zhuzhikov, D.P. (2001). Compartmentalization of proteinases and amylases in Nauphoeta cinerea midgut. Archives of Insect Biochemistry and Physiology, 48, 206-216.
Ferreira, C. & Terra, W.R. (1983). Physical and kinetic properties of a plasma-membrane-bound P-Dglucosidase (cellobiase) from midgut cells of an insect (Rhynchosciara americana larva). Biochemistry Journal, 213, 43-51.
Fossati, P. & Prencipe, L. (1982). Serum triglycerides determined colorimetrically with an enzyme that produces hydrogen peroxide. Clinical Chemistry, 28, 2077-2080.
Hamshou, M., Smagghe, G. & Van Damme, E.J.M. (2010b). Entomotoxic effects of fungal lectin from Rhizoctonia solani towards Spodoptera littoralis. Fungal Biology, 114, 34-40.
Hamshou, M., Smagghe, G., Shahidi-Noghabi, S., De Geyter, E., Lannoo, N. & van Damme, E. J. M. (2010a). Insecticidal properties of Sclerotinia sclerotiorum agglutinin and its interaction with insect tissues and cells. Insect Biochemistry and Molecular Biology, 40, 883-890.
Johnson, N. F. & Triplehorn, C. A. (2004). Borror and DeLong’s introduction to the study of insects. 7th ed. Belmont, CA: Thomson/Brooks Cole publisher. 866 pp.
Kaplan, L.A. & Pesce, A.J. (1996). Clinical Chemistry - Theory Analysis and Correlation. Mosby-Year Book, MO.
Khanjani, M. (2005). Vegetable pests in Iran. Bu’Ali Sina university press. Pages 281-285.
King, J. (1965). The dehydrogenases or oxidoreductases. Lactate dehydrogenase. In: Van Nostrand, D. (Ed.), Practical Clinical Enzymology. Elsevier, London, pp. 83-93.
Komathe, S. S., Kavitha, M., & Swamy, M. J. (2006). Beyond carbohydrate binding: new directions in plant lectin research. Organic and Biomolecular Chemistry, 4, 973-988.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193, 265-275.
Macedo M.L.R., Freire M.G.M., Silva M.B.R. & Coelho L.C.B.B. (2007). Insecticidal action of Bauhinia monandra leaf lectin (BmoLL) against Anagasta kuehniella (Lepidoptera: Pyralidae), Zabrotes subfasciatus and Callosobruchus maculatus (Coleoptera: Bruchidae). Comparative Biochemistry and Physiology Part A, 146, 486-498.
Oppert, B., Kramer, K.J. & McGaughey, W.H. (1997). Rapid microplate assay of proteinase mixtures. Biotechniques, 23, 70-72.
Pinto, P.V.A., Kaplan, A. & Dreal, P.A. (1969). Aldolase: I. Colorimetric determination. Clinical Chemistry, 15, 349-360.
Ramzi Jahroumi, S. (2014). Effect of Citrullus colocynthis L. agglutinin on demographic and physiological characteristics of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae) and gene expression of caspase. Ph. D dissertation in Faculty of Agricultural Sciences, University of Guilan. Pages 95-98.
Ramzi S., Sahragard A., Sendi J.J. & Aalami A. (2013). Effects of an extracted lectin from Citrullus colocynthis L. (Cucurbitaceae) on survival, digestion and energy reserves of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae). Frontiers in Physiology, 12 (4), 328. DOI: 10.3389/fphys.2013.00328.
Ramzi, A., Sahragard, A. & Zibaee, A. (2014) Effects of Citrullus colocynthis agglutinin on intermediary metabolism of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae). Journal of Asia-Pacific Entomology, 17, 273-279.
Ramzi, S. & Sahragard, A. (2013). A lectin extracted from Citrullus colocynthis L. (Cucurbitaceae) inhibits digestive α-amylase of Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae). Journal of Entomological and Acarological Research, 45, 110-116.
Schaefer, E.J. & McNamara, J. (1997). Overview of the diagnosis and treatment of lipid disorders. In: Rifia, N., Warnick, G.R., Dominiczak, M.H. (Eds.), Handbook of lipoprotein testing. AACC Press, Washington, pp. 25-48.
Scriber, J. M. & Slansky Jr F. (1982). The nutritional ecology of immature insects. Annual Review of Entomology, 26, 183-211.
Senthil Nathan, S., Chunga, P.G. & Muruganb, K. (2006). Combined effect of biopesticides on the digestive enzymatic profiles of Cnaphalocrocis medinalis (Guenee) (the rice leaffolder) (Insecta: Lepidoptera: Pyralidae). Ecotoxicology and Environmental Safety, 64, 382-389.
Sorkhabi-Abdolmaleki, S. & Zibaee, A. (2013) Secretagogue Mechanism of Digestive Enzyme Secretion in the Midgut of Andrallus spinidens Fabricius (Hemiptera: Pentatomidae). Proceedings of National Academy of Science India, Section B: Biological Sciences, 82, 373-379.
Sulzenbacher, G., Roig-Zambonia, V., Peumans, W.J., Rougé, P., Van Damme, E.J.M. & Bourne, Y. (2010). Crystal structure of the GalNAc/Gal-specific agglutinin from the phytopathogenic ascomycete Sclerotinia sclerotiorum reveals novel adaptation of a b-trefoil domain. Journal of Molecular Biology, 400, 715-723.
Szasz, G. (1976). Reaction-rate method for gamma-glutamyltransferase activity in serum. Clinical Chemistry, 22, 2051-2055.
Tate, S.S. & Meister, A. (1985). Gamma-glutamyl transpeptidase from kidney. Methods in Enzymology, 113, 400-419.
Thomas, L. (1998). Clinical Laboratory Diagnostic, first ed. TH Books Verlasgesellschaft, Frankfurt. 89-94.
Tsujita, T., Ninomiya, H. & Okuda, H. (1989). p-Nitrophenyl butyrate hydrolyzing activity of hormone-sensitive lipase from bovine adipose tissue. Journal of Lipid Research, 30, 997-1004.
Wakefield, M., Fitches, E. C., Bell, H. A., Gatehouse, A. M. R. (2010). The snowdrop lectin Galanthus nivalis agglutinin (GNA) and a fusion protein ButaIT/GNA have a differential affect on a pest noctuid Lacanobia oleracea and the ectoparasitoid Eulophus pennicornis. Physiological Entomology, 35, 334-342.
Zibaee, A.(2012). Digestive enzymes of large cabbage white butterfly, Pieris brassicae L. (Lepidoptera: Pieridae) from developmental and site of activity perspectives. Italian Journal of Zoology, 79,13-26.
Zibaee, A., Alborzi, Z., Karimi-Malati, A. & Salimi, M. (2014) Effects of a lectin from Polygonum persicaria L. on Pieris brassicae L. (Lepidoptera: Pieridae). Journal of Plant Protection Research 54, 250-257.
Zibaee, A., Karimi-Malati, A. & Janghorbani, A. (2015). In Vitro Interaction of Digestive a-Amylase from Pieris brassicae (Lepidoptera: Pieridae) with a Lectin Extracted from Polygonum persicariae (Polygonaceae). Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. DOI 10.1007/s40011-014-0337-4.