اثر نوع والد و جنس کرم ابریشم بر بیان ژن HSP70 در پاسخ به تنش شدید حرارتی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد گروه علوم دامی، دانشکده علوم کشاورزی، دانشگاه گیلان

2 استادیار گروه علوم دامی و گروه پژوهشی کرم ابریشم، دانشکده علوم کشاورزی، دانشگاه گیلان

3 دانشیار گروه بیوتکنولوژی، دانشکده علوم کشاورزی، دانشگاه گیلان

چکیده

به منظور بررسی تفاوت‌­های ژنتیکی اعم از نوع والد و جنسیت بر پاسخ لارو کرم ابریشم به افزایش دما، بیان پروتئین شوک حرارتی 70 (HSP70) در دو جنس نر و ماده در چهار والد 110 و 104 (واریته­های چینی) و 103 و 107 (واریته­‌های ژاپنی) اندازه­گیری شد. لاروها در روز چهارم از سن پنجم لاروی به مدت 35 دقیقه در معرض شوک حرارتی C45 قرار گرفته و سپس نمونه­گیری از بافت چربی در صفر، دو، چهار و بیست و چهار ساعت پس از شوک حرارتی انجام شد. پس از استخراج RNA و اندازه‌گیری میزان بیان نسبی ژن (qRT-PCR) مشخص شد بیان ژن در جنس ماده 8/13 برابر جنس نر بوده و این تفاوت از نظر آماری بسیار معنی­دار بود (0001/0P<). لذا مقایسه والدها در چهار نوبت نمونه­گیری به تفکیک جنس با استفاده از تجزیه فاکتوریل و آزمون دانکن انجام شد. تفاوت‌های بیان ژن در واریته‌های مورد بررسی بسیار معنی دار بود (001/0P<). در جنس ماده، واریته­ 107 بیشترین بیان ژن HSP70 را با 552 واحد و در جنس نر، واریته‌ 110 با 53 واحد نشان داد. بیش­ترین بیان ژن دو ساعت پس از شوک حرارتیمشاهده شد لیکن با زمان صفر تفاوت معنی­داری نداشت. آزمایش تحمل حرارتی نشان داد که دو والد 110 و 107 (63/0 و 44/0) نسبت به دو والد 103 و 104 (94/0 و 74/0) تلفات کمتر و تحمل حرارتی بیشتری داشتند. به‌طور کلی واریته 103 که پایین­ترین میزان بیان ژن HSP70را داشت به میزان زیادی به شوک حرارتی حساس بود.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effect of parent type and sex on HSP70 gene expression of silkworm in response to severe heat shock

نویسندگان [English]

  • N. Fazeli Moghaddam 1
  • S. H. Hosseini Moghaddam 2
  • M. M. Souhani 3
1 MSc student, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Iran
2 Assistant Professor, Department of Animal Science and Department of Sericulture, Faculty of Agricultural Sciences, University of Guilan, Iran
3 Associate Professor, Department of Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Iran
چکیده [English]

In order to study the parents and sexes effect as genetic elements on response of silkworm larva to increased temperature, the expression of HSP70 was measured in both sexes of four silkworm parents including 110 , 104 (Chinese varieties) and 103, 107 (Japanese varieties). Larvae was exposed to 35 min of heat shock at 45°C and then sampling of larval fat body was done at zero, two, four and twenty-four hours after heat shock in the fourth day of the fifth instar. After RNA isolation and determination of HSP70 relative gene expression by qRT-PCR, results showed that the gene expression in female was 13.8 times of male larvae with high significant differences (P<0.0001). Therefore, the comparison of silkworm parents in the four times samplings were done by factorial analysis and Duncan’s test in each sex separately. HSP70 gene expression differences among varieties was highly significant (P<0.0001). 107 and 110 varieties showed the highest expression of HSP70 gene (552 and 53) among female and male silkworm larvae groups, respectively. This gene was increasingly expressed immediately after heat exposure, but there were no significant differences between the times of zero and 2 hour after heat shock. Thermal tolerance test showed that two varieties of 110 and 107 (0.63 and 0.44) had less mortality and more tolerant than 103 and 104 (0.94 and 0.74). The results showed that the 103 variety which had the lowest HSP70 expression, was also so sensitive to heat stress.

کلیدواژه‌ها [English]

  • Gene Expression
  • HSP70 gene expression
  • Thermotolerance
  • Silkworm parents
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