شناسایی بیوانفورماتیکی ژن‏های القاء شده در میگوی ژاپنی (Marsupenaeus japonicus) در پاسخ به تزریق dsRNA غیراختصاصی با طول بلند و ارتباط آن‏ها با ایمنی علیه ویروس سندروم لکه سفید (WSSV)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 استادیار، پژوهشکده میگوی کشور، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی (AREEO)، بوشهر، ایران.

2 استادیار گروه علوم دامی، دانشکده کشاورزی، دانشگاه شهرکرد، شهرکرد، ایران

10.22103/jab.2025.24530.1638

چکیده

چکیده
هدف: توسعه پایدار صنعت میگوی پرورشی در معرض تهدید طیف وسیعی از عوامل بیماری‏زای مهم از جمله ویروس سندرم لکه سفید (WSSV) قرار دارد که موجب ضررهای اقتصادی شدیدی به این صنعت می‏شود. هدف از این مطالعه، شناسایی ژن‏های افتراقی بیان شده (DEGs) در میگوی کروما یا ژاپنی (Marsupenaeus japonicus) در پاسخ به تزریق dsRNA غیراختصاصی با طول بلند و ارتباط آن‏ها با القای ایمنی علیه بیماری لکه سفید (WSD) بود.
مواد و روش‏ها: هشت داده ‏ریزآرایه مربوط به هموسیت میگوی‏های تزریق شده با PBS و dsRNA غیراختصاصی با طول بلند (از هر گروه 4 نمونه) در فواصل زمانی 24 و 48 ساعت پس از تزریق، از پایگاه داده‌ GEO با شماره دسترسی GSE61541 استخراج و با استفاده از ابزار GEO2R آنالیز شدند. داده‌ها پس از نرمال‏سازی خودکار، به نرم‌افزار اکسل وارد و ژن‌های افتراقی بیان شده (DEGs) با P-Value تصحیح شده کمتر از 05/0 مشخص شدند. ژن‏هایی با LogFC بزرگتر از 2 و کوچکتر از 2- به ترتیب به عنوان ژن‏های با بیان بالا و پایین در نظر گرفته شدند. سپس ژن‌های افتراقی بیان شده مشترک در دو زمان 24 و 48 ساعت پس از تزریق، با استفاده از ابزار VENNY 2.0.2 شناسایی شد.
یافته‏ها: بر اساس تحلیل داده‌های ریز‌آرایه، در مجموع 160 ژن با بیان افتراقی (شامل 111 ژن با بیان بالا و 49 ژن با بیان پایین) در 24 ساعت پس از تزریق dsRNA و 206 ژن با بیان افتراقی (شامل 138 ژن با بیان بالا و 68 ژن با بیان پایین) در 48 ساعت پس از تزریق شناسایی شدند. تحلیل ژن‌های افتراقی با استفاده از ابزار VENNY نشان داد که 74 ژن مشترک (شامل 67 ژن با افزایش بیان و 7 ژن با کاهش بیان) بین دو زمان 24 و 48 ساعت پس از تزریق dsRNA وجود داشت. تزریق dsRNAهای غیراختصاصی با طول بلند توانست موجب القای بسیاری از ژن‌های مرتبط با ایمنی ذاتی نظیر Ribonuclease T2، C-type lectin 2، HSP90، Caspase Nc-like و TRIM64 در پاسخ به WSSV در میگوهای ژاپنی گردد.
نتیجه‏گیری: فعالیت‏های مختلف زیستی نظیر تشخیص الگوهای مولکولی مرتبط با عامل بیماری، اتصال، حفظ هموستازی، آپوپتوزیس، کاتالیزوری، انتقال غشایی، چپرونی و RNAi برای ژن‏های القاء شده در میگوی ژاپنی در پاسخ به تزریق dsRNA غیراختصاصی با طول بلند پیش بینی شد. شناسایی این ژن‏‏ها می‏تواند در جهت یافتن نشانگر‏های زیستی مرتبط با بیماری لکه سفید در میگو، تشخیص و یا طراحی مهارکننده‏ علیه آن مفید باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Bioinformatics identification of induced genes in Marsupenaeus japonicus in response to injection of long non-specific dsRNA and their association with immunity against white spot syndrome virus (WSSV)

نویسندگان [English]

  • Reza Pasandideh 1
  • Majid Pasandideh 2
1 Assistant Professor, Iranian Shrimp Research Center, Iranian Fisheries Science Research Institute, Agricultural Research, Education & Extension Organization (AREEO), Bushehr, Iran.
2 Assistant Professor, Department of Animal Science, Faculty of Agriculture, Shahrekord University, Shahrekord, Iran
چکیده [English]

Abstract
Objective
The sustainable development of the farmed shrimp industry is threatened by a wide range of important pathogens, including white spot syndrome virus (WSSV), which causes severe economic losses to the industry. The aim of this study was to identify differentially expressed genes (DEGs) in Kuruma shrimp (Marsupenaeus japonicus) in response to injection of long non-specific dsRNA and their association with survival and immunity against white spot disease (WSD).
Materials and methods
Eight microarray data from shrimp hemocytes injected with PBS and long non-specific dsRNA (four samples from each group) at time intervals of 24 and 48 hours after injection were extracted from the GEO database with accession number GSE61541 and analyzed using the GEO2R tool. After automatic normalization, the data were imported into Excel software and differentially expressed genes (DEGs) were identified with an adjusted P-Value less than 0.05. Genes with LogFC greater than 2 and less than -2 were considered as up- and down-expressed genes, respectively. Then, the commen DEGs were identified using the VENNY 2.0.2 tool.
Results
Data analysis showed that 160 DEGs were identified in 24 hours after dsRNA injection, of which 111 genes were up-regulated and 49 genes down-regulated. As well as, the results showed that 206 DEGs were identified in 48 hours after dsRNA injection, of which 138 genes were up-regulated and 68 genes down-regulated. The analysis of DEGs using the VENNY tool showed that 74 common genes were identified in 24 and 48 hours after dsRNA injection, of which 67 genes were up-regulated and 7 genes down-regulated. Shrimp injected with dsRNA had a higher number of DEGs than the PBS-injected group, most of which were up-regulated. The results of this study confirmed that injection of long non-specific dsRNAs could induce many genes related to innate immunity, such as Ribonuclease T2, C-type lectin 2, HSP90, Caspase Nc-like, and TRIM64 in response to WSSV infection in Kuruma shrimp.
Conclusions
Various biological functions were predicted for genes induced in Kuruma shrimp in response to injection of long non-specific dsRNA included recognition of pathogen-associated molecular patterns (PAMPs), binding, homeostasis, apoptosis, catalytic, transmembrane, chaperone and RNAi. Identification of these genes could be useful in finding markers associated with white spot infection, diagnosis, or designing inhibitors against that disease.
Keywords: Innate immunity, RNA interference (RNAi), Microarray, Kuruma shrimp (Marsupenaeus japonicas), White spot syndrome virus (WSSV)

کلیدواژه‌ها [English]

  • Innate immunity
  • RNA interference (RNAi)
  • Microarray
  • Kuruma shrimp (Marsupenaeus japonicas)
  • White spot syndrome virus (WSSV)
شیری نیما، سوری مرتضی، صفدریان بهزاد (1400) مقایسه میگوی ژاپنی (Marsupenaeus japonicus) و پاسفید غربی (Litopeanaues vannamei)  از نظر معیارهای تکثیر و پرورش با رویکرد امکان سنجی. مجله ترویجی میگو و سخت پوستان، 6 (2)، 12-4.
 
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