بهینه‌سازی محیط کشت در تولید اسپور و کریستال باکتری Bacillus thuringiensis subsp. kurstaki و ارزیابی زهرآگینی آن روی لارو سن دوم پروانه Ephestia kuehniella

نوع مقاله : مقاله پژوهشی


1 دانشجوی دکتری حشره‌شناسی، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه

2 دانشیار گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران

3 . استاد گروه صنایع غذایی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه

4 استاد گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه

5 دانشیار بخش کنترول بیولوژیک، سازمان تحقیقات، آموزش و ترویج کشاورزی، تهران گیاهپزشکی کشور

6 استادیار بخش تحقیقات گیاهپزشکی، سازمان تحقیقات کشاورزی و منابع طبیعی استان آذربایجان غربی، ارومیه


در پژوهش حاضر بیمارگری اسپور و کریستال باکتری (HD-1) Bacillus thuringiensis subsp. kurstaki تولیدشده در انواع محیط کشت در شرایط تخمیری و غیر‌تخمیری روی سن دوم لاروی Ephestia kuehniella L. بررسی شد. تعداد کلنی‌، تعداد اسپور، ضریب شکست نوری (OD) و وزن خشک سلولی در محیط کشت غنی‌شده و غنی‌نشده در شرایط تخمیری پس از 96 ساعت کشت به‌ترتیب (1014‌×2/28 CFU برمیلی‌لیتر،‌1010×‌8/11اسپوربرمیلی‌لیتر، 108‌×‌0/28‌میلی‌گرم-برمیلی‌لیتر، 0/11گرم‌برلیتر) و (1014‌×6/26 CFU برمیلی‌لیتر،‌1010×‌0/10 اسپور‌بر‌میلی‌لیتر، 108‌×‌4/24 میلی‌گرم‌برمیلی‌لیتر، 9/9 گرم‌برلیتر) و در شرایط تخمیر‌نشده (1014‌×1/18CFU بر‌میلی‌لیتر،1010×‌1/8 اسپوربر‌میلی‌لیتر، 108‌×‌4/16 میلی‌گرم‌بر میلی‌لیتر، 9/8 گرم‌برلیتر) و (1014‌×8/16CFU‌ بر‌میلی‌لیتر،‌1010×‌4/7 اسپوربرمیلی‌لیتر، 108‌×‌6/15‌میلی‌گرم‌برمیلی‌لیتر، 9/7 گرم‌برلیتر) به دست آمد. شاخص‌LC‌50 سوسپانسیون اسپور و کریستال رشد کرده در محیط کشت غنی‌شده و غنی‌نشده در شرایط تخمیر‌شده در مدت 96 ساعت کشت، پس از دو روز تیمار لاروها (9/0 و 0/3) و در شرایط تخمیر‌نشده (5/1 و 2/4) میکروگرم‌برمیلی‌لیتر و شاخصLT50 در شرایط تخمیر‌شده (2/6 و 7/10) و در شرایط تخمیر‌نشده (2/7 و 6/12) ساعت برآورد شد. بررسی‌های زیست‌سنجی میزان مرگ‌ومیر حاصل از سوسپانسون رشد کرده در محیط کشت غنی‌شده و غنی‌نشده در شرایط تخمیر‌شده پس از 96 ساعت کشت، پس از دو روز تیمار لاروها به ترتیب (0/61 و 6/46) و در در شرایط تخمیر‌نشده (3/53 و 6/38) درصد نشان داد. نتایج نشان داد محیط کشت غنی‌شده در شرایط تخمیر‌شده سبب تولید بیشترین میزان اسپور و کریستال و در نتیجه افزایش زهرآگینی باکتری روی لارو سن دوم پروانه E. kuehniellaشد.


عنوان مقاله [English]

Improvement the media culture for spore/crystal production of Bacillus thuringiensis subsp. kurstaki and evaluate their virulence against second larval instar of Ephestia kuehniella L.

نویسندگان [English]

  • Elmira Mokhtarnejhad 1
  • Shahram Aramideh 2
  • Mahmood Rezazad Bari 3
  • Mohammad Hassan Safaralizadeh 4
  • Mohammad Reza Rezapanah 5
  • Maryam Forouzan 6
1 PhD Student of Entomology, Faculty of Agriculture, Urmia University, Urmia
2 Associate Professor of Plant Protection Department, Faculty of Agriculture, Urmia University, Urmia, Iran
3 Professor of Food Science Department, Faculty of Agriculture, Urmia University, Urmia
4 Professor of Plant Protection Department, Faculty of Agriculture, Urmia University, Urmia
5 Associate Professor of Department of Biological Control, Iranian Research Institute of Plant Protection, Agriculture Research Education and Extension Organization, Tehran
6 Plant Protection Research Department. West Azarbaijan, Agriculture and Natural Resources Research Center, AREEO, Urmia
چکیده [English]

In the present study, the pathogenicity of spores/crystals produced byBacillus thuringiensis subsp. kurstaki (HD-1) in different culture media within fermented and non fermented conditions were determined against the second larval instar of Ephestia kuehniella L. The colony forming unit (CFU), spore count, optical density (OD) and cell mass indices in enriched and non-enriched liquid media of fermented condition after 96 h cultivation were (28.2‌×‌1014 CFU.mL‌-1, 11.8‌×‌1010 spore.mL‌-1, 28.0‌×‌108 mg.mL-1 and 11.0 g.L-1) and (26.6‌×‌1014 CFU.mL‌-1, 10.0‌×‌1010 spore.mL‌-1, 24.4‌×‌108 mg.mL-1 and 9.9 g.L-1), respectively while those were found to be (18.1‌×‌1014 CFU.mL‌-1, 8.1‌×‌1010 spore.mL‌-1, 16.4‌×‌108 mg.mL-1 and 8.9 g.L-1) and (16.8‌×‌1014 CFU.mL‌-1, 7.4 ×‌1010 spore.mL -1, 15.6‌×‌108 mg.mL-1 and 7.9 g.L-1) in non-fermented condition, respectively. The LC50 value of Btk spore/crystal cultured on enriched and non-enriched liquid media for 96h cultivation in fermented condition after 2 days treatment against larval instar were obtained (0.9 and 3.0) and in non-fermented condition (1.5 and 4.2) µg.mL-1, respectively.The LT50 value in fermented condition (6.2 and 10.7) and in non-fermented condition (7.2 and 12.6) h were, respectively. Bioassay studies indicated that the Btk spore/crystal suspention cultured on enriched and non-enriched media in fermented condition for 96h cultivation after 2 days treatment had (61.0 and 46.6-%) and in non-fermented condition (53.3 and 38.6%) mortality, respectively. The results confirm that enriched culture medium in fermented conditions produced the highest amount of spores and crystals and thus increased bacterial toxicity on the second larval instar of E. kuehniella after treatment.

کلیدواژه‌ها [English]

  • Bacillus thuringiensis subsp. kurstaki
  • Ephestia kuehniella
  • Fermented condition
  • Enriched medium
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