The phylogenetic reconstruction of biosynthesizing genes from Cyanobacteria isolated from Ziarat waterfall of Golestan Province

Document Type : Research Paper

Authors

Department of Biotechnology, Faculty of Converging Sciences and Technologies, Islamic Azad University, Science and Research, Tehran, Iran

10.22092/ijb.2023.362834.1418

Abstract

Cyanobacteria are anaerobic and photosynthesizing prokaryotes. In the past, the classification of cyanobacteria was only based on morphological characteristics. Today, advanced techniques such as molecular markers are used to achieve more accurate and reliable classification. In this research, sampling was done from the Ziarat waterfall in Golestan province. After cultivation and purification in solid culture medium BG-11, molecular identification of natural compounds’ biosynthesizing genes (hassallidins synthetase (hasN) and geosmin A (geoA) along with divergence analyses were performed using the amplification of the chloroplast genes RNA polymerase C1 (rpoC1).
Then, the strain containing the above genes was identified by amplification of 16S rRNA and internal transcribed spacer (ITS) genes. Phylogenetic trees were built using the Maximum Likelihood method and the appropriate model with the help of a web server on the IQ-Tree server. The secondary structure of ITS was drawn in different parts of helix D1-D1′, D2, D3, tRNAIle, tRNAAla, BOX B, BOX A, and V3 using the Mfold program. The results showed that only the Nodosilinea sp. 1359 (Leptolyngbyaceae, Synechococcales) strain has the genes mentioned above. In addition, the results of calculating the KA/KS of hasN and geoA genes and the phylogenetic incongruence of 16S rRNA and rpoC1 genes showed that natural selection by creating positive mutations has led to diversity in the studied strain. This study is among the first research conducted on the molecular phylogeny of cyanobacteria producing natural compounds in Ziarat waterfall.

Keywords

Article Title [Persian]

بازسازی فیلوژنتیک ژن‌های بیوسنتزکننده سیانوباکتری‌های جدا شده از آبشار زیارت استان گلستان

Authors [Persian]

  • بهاره نوروزی
  • سمانه جعفری پرزانی
گروه بیوتکنولوژی، دانشکده علوم و فناوری‌های همگرا، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران
Abstract [Persian]

سیانوباکتری‌ها، پروکاریوت‌های بی‌هوازی و فتوسنتز کننده هستند. در گذشته رده‌بندی سیانوباکتری‌ها تنها براساس صفات مورفولوژیکی استوار بود. اما امروزه از تکنیک‌های پیشرفته مانند نشانگرهای مولکولی برای دستیابی به رده‌بندی دقیق‌تر و قابل اعتمادتر استفاده می‌شود. در این تحقیق نمونه‌برداری از آبشار زیارت استان گلستان انجام شد. پس از کشت و خالص‌سازی در محیط کشت جامد BG-11، شناسایی مولکولی ژن‌های بیوسنتزکننده ترکیبات طبیعی (hasN، و geoA) به همراه آنالیزهای واگرایی با تکثیر ژن‌های rpoC1 انجام گردید. سپس سویه حاوی ژن‌های فوق با تکثیر ژن‌های 16S rRNA و ITS شناسایی گردید. درختان فیلوژنتیک با استفاده از روش بیشینه درست‌نمایی و مدل مناسب به کمک وب سرور IQ-Tree ساخته شد. ساختار ثانویه ITS، در بخش‌های مختلف هلیکس D1-D1′، D2، D3، tRNAIle، tRNAAla، BOX B، BOX A و V3 به کمک برنامه M-fold رسم شد. نتایج نشان داد که تنها سویه Nodosilinea sp. 1359 دارای ژن‌های فوق بوده و متعلق به تیره Leptolyngbyaceae و راسته Synechococcales است. علاوه بر آن، نتایج حاصل از محاسبه میزان KA/KS ژن‌های hasN و geoA و ناسازگاری داده‌های ژنتیکی 16S rRNA و rpo C1 نشان داد که انتخاب طبیعی با ایجاد جهش‌های مثبت منجر به ایجاد تنوع در سویه مورد مطالعه شده است. این مطالعه، جزو نخستین تحقیقات انجام شده روی فیلوژنی مولکولی سیانوباکتریوم تولیدکننده ترکیبات طبیعی در آبشار زیارت است.

Keywords [Persian]

  • Nodosilinea sp. 1359
  • 16S rRNA
  • ITS
  • hasN
  • geoA
  • rpoC1
  • ژنهای بیوسنتز کننده ترکیبات طبیعی

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The Iranian Journal of Botany
Volume 29, Issue 2
December 2023
Pages 187-200

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History

  • Receive Date: 08 July 2023
  • Revise Date: 12 October 2023
  • Accept Date: 03 November 2023

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