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
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