پاسخ‌های ایمنی سن شکارگر (Andrallus spinidens Fabricius (Hem.: Pentatomidae به قارچ بیمارگر Beauveria bassiana Boisdual

نوع مقاله: مقاله پژوهشی

نویسندگان

1 گروه گیاه‌پزشکی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت-ایران

2 گروه زراعت و اصلاح نباتات، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت-ایران

3 بخش کنترل بیولوژیک، موسسه تحقیقات گیاه‌پزشکی، آمل-ایران

چکیده

در این پژوهش، تاثیر دو جدایه از قارچ بیمارگر Beauveria bassiana Boisduval بر سامانه ایمنی افراد بالغ سن شکارگر Andrallus spinidens Fabricius (Hemiptera: Pentatomidae) مورد بررسی قرار گرفت. چهار نوع سلول خونی در همولنف حشره بالغ شناسایی شد که شامل پروهموسیت‌ها، پلاسموتوسیت‌ها، گرانولوسیت‌ها و اوونوسیت‌ها بودند. همچنین تاثیر دو جدایه قارچ  B. bassiana و لاتکس­بید روی ایمنی سلولی و فعالیت فنل اکسیداز مورد بررسی قرار گرفت. بیشترین تعداد سلول­ خونی شمارش شده به­ترتیب 48، 48، 12 و 12 ساعت پس از آلودگی، در توئین (شاهد)، لاتکس بید، جدایه BB2 و جدایه AM-118 به­ دست آمد. بیشترین تعداد پلاسماتوسیت ­ها نیز در 48، 48، 24و 3 ساعت پس از آلودگی در توئین (شاهد)، لاتکس بید، جدایه BB2 و جدایه AM-118 و بیشترین تعداد گرانولوسیت ­ها نیز به ­ترتیب در 48، 48، 12 و 12 ساعت پس از آلودگی مشاهده شد. بیشترین میزان گره نیز به ­ترتیب در 48، 48، 12 و6  ساعت پس از آلودگی و در مورد فعالیت فنل اکسیداز هم بیشترین فعالیت 6 ساعت پس از آلودگی در توئین، جدایه BB2 و جدایه AM-118 و 3 ساعت پس از آلودگی برای لاتکس ­بید مشاهده شد. جهت تعیین ژن تولید کننده پیش فنل اکسیداز، RNA سلول­های خونی استخراج شده و پس از سنتز cDNA، ژن مذکور با استفاده از یک جفت آغازگر رفت و برگشت دژنره تکثیر شد. نتیجه نشان ­دهنده محصولی با 675 جفت باز بر مبنای خوانش برگشت بود که با پیش ­فنل اکسیداز سن Anasa tristis De Geer  تشابهی 66 درصدی داشت. نتایج این پژوهش نشان می­ دهند که پاسخ­ های ایمنی سن شکارگر به جدایه ­های مختلف قارچ B. bassianaمتفاوت است، بنابراین در برنامه­ های مدیریت تلفیقی باید از جدایه ­هایی استفاده کرد که برای شکارگر بیمارگری کمتری داشته باشند.

کلیدواژه‌ها


عنوان مقاله [English]

Immune responses of Andrallus spinidens Fabricius (Hem.: Pentatomidae) to the entomopathogenic fungus, Beauveria bassiana Boisdual

نویسندگان [English]

  • H. Firouzbakht 1
  • A. Zibaee 1
  • M. M. Sohani 2
  • H. Hoda 3
1 Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht-Iran
2 Assistant Professor, Department Agronomy and Plant breeding, Faculty of Agricultural Sciences, University of Guilan, Rasht-Iran
3 Biological Control Department, National Institute of Plant Protection, Amol, Iran
چکیده [English]

In this study, the effects of two isolates from the entomopathogenic fungus, Beauveria bassiana Boisduval were determined in immune system of the predatory hemipteran, Andrallus spinidens Fabricius (Hemiptera: Pentatomidae). Four types of hemocytes were identified in the hemolymph of the adult bug including prohemocytes, plasmatocytes, granulocytes and oenocytoids. Also, the effects of two entomopathogenic fungus isolates and latex bead were studied on the activity of phenoloxidase. The highest number of total hemocytes were found at time intervals of 12, 12, 48 and 48 hours post-injection by Tween80 (Control), latex bead, BB2 and AM-118, respectively. The highest number of plasmatocytes were determined 48, 48, 24 and 3 hours post-injection by Tween80 (Control), latex bead, BB2 and AM-118 but the number of granulocytes were observed after 48, 48, 12 and 12 hours, respectively. The highest number of nodules were observed 48, 48, 12 and 6 hours post-injection Tween80 (Control), latex bead, BB2 and AM-118, respectively. The highest activity of phenoloxidase were determined 6 hours post-injection by Tween80 (Control), BB2 and AM-118 and 3 hours post-injection by Tween80, respectively. To determine the gene of proPhenoloxidase, RNAs of the hemocytes were extracted and it was amplified by a pair of degenerate forward and reverse primers followed by cDNA synthesis. The results revealed a product with 675 bp based on reverse sequencing which had a 66% similarity with prophenoloxidase of Anasa tristis De Geer.Results of this study demonstrate that immune responses of the predacious hemipteran are different regarding various isolates of B. bassiana so the isolates should be used which have the lower virulence against the predator in integrated pest management programs.

کلیدواژه‌ها [English]

  • Andrallus spinidens
  • Beauveria bassiana
  • Cellular immunity
  • Phenoloxidase
Ashida, M. and Brey, P. 1997. Recent advances in research on the insect prophenoloxidase cascade. In Brey, P. and Hultmark, D. (Eds.). Molecular Mechanisms of Immune Responses in Insects. Chapman and Hall, London. UK,.pp, 135-171.

Azambuja, P., Garcia, E. S. and Ratcliffe, N. A. 1991. Aspects of classification of hemiptera hemocytes from six triatomine species. Memórias do Instituto Oswaldo Cru 86: 1-10.

Borges, A., Santos, P., Furtado, A. and Figueiredo, R. 2008. Phagocytosis of latex beads and bacteria by hemocytes of the triatomine bug Rhodnius prolixus (Hemiptera: Reduvidae). Journal of Micron 39: 486-494.

Cerenius, L., Lee, B. L. and Söderhäll, K. 2008. The proPO-system: pros and cons for its role in invertebrate immunity. Journal of Trends in Immunology 29: 263-271.

Firouzbakht, H., Zibaee, A., Hoda, H. and Sohani, M. M. 2015. Virulence Determination of Beauveria bassiana Isolates on a Predatory Hemipteran, Andrallus spinidens Fabricius (Hemiptera: Pentatomidae). Acta Phytopathologica et Entomologica Hungarica 50: 115–126.

Franssens, V., Smagghe, G., Simonet, G., Claeys, I., Breugelmans, B., DeLoof, A. and Vanden Broeck, J. 2006. 20- Hydroxy ecdysone and juvenile hormone regulate the laminarin- induced nodulation reaction in larvae of the fleshfly, Neobellieria bullata. Developmental and Comparative Immunology 30: 735–740.

Gardiner, E. M. M. and Strand M. R. 2000. Hematopoiesis in larval Pseudoplusia includens and Spodoptera frugiperda. Archive of Insect Biochemistry and Physiology 43: 147-164.

Gillespie, J. P. and Khachatourians, G. G. 1992. Characterization of the Melanoplus sanguinipes haemolymph after infection with Beauveria bassiana. Comparative Biochemistry and Physiology-Part B103: 455-463.

Gillespie, J. P., Burnett, C. and Charnley, A. K. 2000. The immune response of the desert locust Schistocerca gregaria during mycosis of the entomopathogenic fungus, Metarhizium anisopliae var acridum. Journal of Insect Physiology 46: 429-437.

Gupta, A. 1985. Cellular elements in the hemolymph. In Gilbert, L. Comprehensive insect physiology, biochemistry and pharmacology. Acdemic press, NY, USA. pp 401-451.

Hazarika, L. K. and Gupta A. P. 1987. Variation in hemocyte populations during various developmental Stages of Blattela germanica (L.) (Dictyoptera, Blattellidae). Zoological Science 4: 307-313.

Hung, S. Y. and Boucias, D. G. 1992. Influence of Beauveria bassiana on the cellular defense response of the beet armyworm, Spodoptera exigua. Journal of Invertebrate Pathology60: 152-158.

Lavine, M. and Strand, M. 2002. Insect hemocytes and their role in immunity. Insect Biochemistry and Molecular Biology32: 1295-1309.

Leonard, C., Kenneth, S. and Ratcliffe, N. A. 1985. Studies on prophenoloxidase and protease activity of Blaberua craniifer haemocytes. Insect Biochemistry 15: 803-810.

Ling, E. and Yu, X. Q. 2005. Prophenoloxidase binds to the surface of hemocytes and is involved in hemocyte melanization in Manduca sexta. Insect Biochemistry and Molecular Biology 35: 1356-1366.

Lu, M., Du, X., Cao, S. S., Liu, P. and Li, J. 2013. Molecular cloning and characterization of the first caspase in the striped stem borer, Chilo suppressalis. International Journal of Molecular Sciences 14: 10229-10241.

Mohaghegh, J. and Najafi, I. 2003. Predation capacity of Andrallus spinidens (F.) (Hemiptera: Pentatomidae) on Naranga aenescens Moore (Lep.: Noctuidae) under semi-field and field conditions. Applied Entomology and Phytopathology 71: 57-68.

Najafi-Navaee, A., Saeb, H. and Osco, T. 1998.  Biology and ecology of Andrallus spinidens F. as the predator of rice, cotton and maize pests. Proceedings of 13th Iranian Plant Protection Congress. 22-26 August, Iran. pp. 49.

Ramzi, S. and Zibaee, A. 2014. Biochemical properties of different entomopathogenic fungi and their virulence against Chilo suppressalis (Lepidoptera: Crambidae) larvae. Biocontrol Science and Technology 24: 597–610.

Sorkhabi-Abdolmaleki, S., Zibaee, A., Hoda, H., Hosseini, R. and Fazeli-Dinan, M. 2013. Proteolytic compartmentalization and activity in the midgut of Andrallus spinidens Fabricius (Hemiptera: Pentatomidae). Journal of Entomological and Acarological Research 45: 33-41.

Stanley, D. W. and Miller, J. S. 2006. Eicosanoid actions in insect cellular immune functions. Journal of Entomologia Experimentalis et Applicata 119: 1-13.

Zibaee, A., Bandani, A. R., Talaei-Hassanlouei, R. and Malagoli, D. 2011. Cellular immune reactions of Eurygaster integriceps, to the entomopathogenic fungus, Beauveria bassiana and its secondary metabolites. Journal of Insect Science 11: 138.