استفاده از الیسیتورهای مختلف زنده و غیر زنده در کشت سوسپانسیون سلول فندق جهت بررسی تغییرات در بیان یکی از ژن‏های کلیدی در مسیر بیوسنتز تاکسول، 3- N-Debenzoyl-2-Deoxytaxoln-Benzoyltransferase (DBTNBT)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، بخش بیوتکنولوژی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، ایران.

2 بخش بیوتکنولوژی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، ایران.

10.22103/jab.2025.23315.1564

چکیده

چکیده فارسی
مقدمه:
پاکلیتاکسل (تاکسول) که داروی طبیعی موثر شیمی درمانی است، برای درمان طیف وسیعی از سرطان ها استفاده می شود. از نظر تاریخی، سرخدار (Taxus spp.) منبع اصلی تاکسول بوده است. در حال حاضر، فندق (Corylus avellana L.) را می توان به عنوان یک منبع ارزان و در دسترس برای تهیه تاکسول دانست. این مطالعه اثرات احتمالی الیسیتورهای مختلف را بر سطوح بیان ژن DBTNBT (3-N-debenzoyl-2-deoxytaxolN-benzoyltransferase) در کشت های سوسپانسیون سلولی فندق بررسی کرد. ژن DBTNBT یک ژن کلیدی درگیر در مسیر بیوسنتز تاکسول است.
مواد و روشها:
کشت سوسپانسیون سلولی فندق در MS مایع همراه با 2 میلی‏گرم در لیتر 2,4-D و 2/0 میلی‏گرم در لیتر BAP انجام شد. کشت های سلولی فندق با عصاره قارچی Cheatomium globosum در غلظت های 25، 50 و 100 میلی‏گرم در لیتر ، متیل جاسمونات (MeJA) (0، 100 و 200 میکرولیتر) و نیترات نقره (0، 15 و 30 میلی‏گرم در لیتر) تیمار شدند.
نتایج:
نتایج حاکی از اثرات مثبت الیسیتورها بر بیان ژن DBTNBT بود. به طوری که بالاترین سطح بیان ژن DBTNBT در کشت سوسپانسیون سلولی فندق تیمار شده با عصاره قارچی C. globosum در غلظت 50 میلی‏گرم در لیتر مشاهده شد و سطح بیان ژن DBTNBT در این غلظت 75/4 برابر نسبت به شاهد افزایش یافت. به طور مشابه، در کشت سوسپانسیون سلولی فندق با متیل جاسمونات، بیان ژن DBTNBT در غلظت 200 میکرولیتر 6/17 برابر در مقایسه با نمونه های شاهد افزایش یافت. علاوه بر این، هنگامی که غلظت AgNO3 دو برابر گردید و به 30 میلی گرم در لیتر رسید، میزان بیان ژن 14 برابر نسبت به تیمار شاهد افزایش یافت.
نتیجه گیری:
یافته های این مطالعه نشان می دهد بالاترین میزان بیان ژن DBTNBT مربوط به تیمار MeJA است، با وجود اینکه تمام تیمارهای بکار رفته قادر به افزایش موفقیت آمیز بیان ژن DBTNBT در کشت سوسپانسیون سلولی فندق بودند.

کلیدواژه‌ها

عنوان مقاله [English]

Using Various Biotic and Abiotic Elicitors in Hazelnut Cell Suspension Cultures to Investigate the Expression of the 3-N-Debenzoyl-2-DeoxyTaxoln-Benzoyltransferase Gene in the Paclitaxel Biosynthesis Pathway

نویسندگان [English]

  • Raziyeh Bahrasmani Sardoo 1
  • Sara Alsadat Rahpeyma 2
  • Mehdi Mansouri 2
1 MS student of plant biotechnology, Department of Agricultural Biotechnology, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
2 Department of Agricultural Biotechnology, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
چکیده [English]

Background
Paclitaxel (PC) is a naturally occurring chemotherapeutic medication used to treat various malignancies. Hazelnut (Corylus avellana L.) is currently a readily available and affordable Paclitaxel source. In this study, the potential effects of some biotic and abiotic elicitors were investigated on expression levels of the DBTNBT gene (3-N-debenzoyl-2-deoxytaxolN-benzoyltransferase) in cell suspension cultures of C. avellana. The DBTNB gene is one of the key genes in the downstream biosynthesis pathway of Paclitaxel.
Materials and methods
Hazelnut yellow friable calli taken from and often subcultured in MS media, supplemented with 2,4-D (2 mg/L) and BAP (0.2 mg/L), was suspended in liquid MS of the same composition. The elicitors methyl jasmonate (MeJA) (0, 100, and 200 (µL)), silver nitrate (0, 15, and 30 (mg/L)), or fungus extract (0, 25, 50, and 100 (mg/L)) were applied to the hazelnut cell cultures for 48 hours during the middle growth phase.
Results
The present study demonstrated the inductive effects of the elicitors on the expression of the DBTNBT gene. Compared to the control samples, in the C. avellana cell suspension culture, DBTNBT gene expression was affected positively by MeJA, and the most increase in DBTNBT gene expression (17.6-fold) was obtained from the treatment of 200 µL MeJA. The rate of gene expression rose considerably, up to 14 times greater than that of the control, when the concentration of AgNO3 was increased to 30 mg/L. Fungal extract affected DBTNBT gene expression; a suspension culture of C. avellana cells treated with 50 mg/L fungal extracts of C. globosum revealed a 4.75-fold increase in DBTNBT gene expression relative to the reference. However, 100 mg/L of C. globosum extracts reduced gene expression compared to the control.
Conclusions
MeJA had the highest degree of DBTNBT gene expression of any elicitor therapy used in this investigation, however all applied elicitation treatments were able to successfully increase the DBTNBT gene expression in hazelnut cell suspension cultures.

کلیدواژه‌ها [English]

  • Corylus avellana
  • DBTNBT
  • gene expression
  • paclitaxel
  • qRT‒PCR

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مجله بیوتکنولوژی کشاورزی
دوره 17، شماره 3
شهریور 1404
صفحه 327-344

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