نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه علوم باغبانی،دانشکده کشاورزی، دانشگاه شهرکرد

2 استادیار، بخش علوم و مهندسی باغبانی، دانشکده کشاورزی شهرکرد، دانشگاه شهرکرد، شهرکرد، ایران.

3 استادیار، بخش مکانیک بیوسیستم، دانشکده کشاورزی شهرکرد، دانشگاه شهرکرد، شهرکرد، ایران.

4 استادیار، بخش گیاهپزشکی، دانشکده کشاورزی شهرکرد، دانشگاه شهرکرد، شهرکرد، ایران.

5 دانشیار، بخش مکانیک بیوسیستم، دانشکده کشاورزی شهرکرد، دانشگاه شهرکرد، شهرکرد، ایران.

چکیده

در این پژوهش ژن tms2 عامل گال طوقه در دو رقم رز Rosa hybrida L. ("Pearl" و"Angelina" ) با کمک روش­های الکتروشیمیایی شناسایی شد. در این پژوهش تثبیت و هیبریداسیون توالی تک‌رشته (ssDNA) مربوط به ژن tms2، Agrobacterium tumefaciens با روش طیف‌سنجی امپدانس الکتروشیمیایی بررسی شد. الکترود گرافیتی مغز مداد با نانو موادی شامل گرافن اکساید احیاشده و نانوذرات طلا اصلاح شد. زیست­حسگر توانست برای تشخیص نمونه­های آلوده رز "Angelina" و "Pearl" به کار رود. هیبریداسیون DNA/DNA با قراردادن الکترود اصلاح‌شده با ssDNA در غلظت دو میکرومولار DNA هدف انجام شد. نتایج تثبیت و هیبریداسیون مناسب DNA را در سطح الکترود نشان داد. زیست­حسگر طراحی‌شده گزینش‌پذیری خوبی دارد. نانوزیست­حسگر مبتنی برDNA ، دارای مزایایی از جمله هزینه کم، سادگی و قابلیت کوچک‌سازی می­باشد و می­تواند زمینه­ای برای توسعه  ابزار تشخیص ژنومی را فراهم آورد.

کلیدواژه‌ها

عنوان مقاله [English]

Detection of tms2 Gene Causing Crown Gall in Two Commercially Cut Flower Roses ("Pearl" And "Angelina") by a Nano Biosensor

نویسندگان [English]

  • Akram Vatankhah 1
  • Saeed Reezi 2
  • Zahra Izadi 3
  • Abdorahman Motamedi 4
  • Mahdi Ghasemi Varnamkhasti 5

1 Ph.D Student, Department of ,Horticulture faculty of Agriculture, Shahrekord University

2 Assistant Professor, Department of Science and Horticultural Engineering, Faculty of Agriculture, Shahrekord, Shahrekord University, Shahrekord, Iran.

3 Assistant Professor, Department of Biosystem Mechanics, Faculty of Agriculture, Shahrekord, Shahrekord University, Shahrekord, Iran.

4 Assistant Professor, Department of Plant Protection, Faculty of Agriculture, Shahrekord, Shahrekord University, Shahrekord, Iran.

5 Associate Professor, Department of Biosystem Mechanics, Faculty of Agriculture, Shahrekord, Shahrekord University, Shahrekord, Iran.

چکیده [English]

In this study, the tms2 gene, the cause of crown gall, is detected in two Rosa hybrida L. (“Pearl” and “Angelina”) cultivars using electrochemical methods. In this study, stabilization, and hybridization of single-strand DNA (ssDNA) related to tms2 gene, Agrobacterium tumefaciens is investigated by electrochemical impedance spectroscopy. For this purpose, the pencil graphite electrode is modified with nanomaterials including reduced graphene oxide and gold nanoparticles. The biosensor could be used to detect infected samples of “Angelina and “Pearl roses. DNA/DNA hybridization is performed by placing an electrode modified with ssDNA at a concentration of two (µmol/L) target DNA. Results show proper stabilization and hybridization of DNA at the electrode surface. The designed biosensor has good selectivity. DNA-based nanosensor has advantages such as low cost, simplicity, and scalability and can be the basis for the development of genomic detection tools.

کلیدواژه‌ها [English]

  • Plant pathogens
  • biosensors
  • electrochemical methods
  • crown gall
  • Rosa hybrida L
Agrios, G.N. (2005). Plant Pathology, Academic press, Elsevier academic press. New York. 922 p.
Atson, E. )2019(. Use of Amino Oligosaccharins and Alternaria Activated Protein in Management of Crown Gall and Enhancement of Growth in Roses. M.Sc. Thesis. Department of Plant Science and Crop Protection. College of Agriculture and Veterinary Sciences. University of Nairobi. 86p.
Azadi, P., Beyrai zadeh, E., & Otang ntui, V. (2013). A simple protocol for somatic embryogenesis in Rosa hybrida L. cv. Apollo. The Journal of Horticultural Science and Biotechnology, 88(4), 399-402.
Bhatt, N.; Huang, P.J.J.; Dave, N., & Liu, J. )2011(. Dissociation and degradation of thiol-modified DNA on gold nanoparticles in aqueous and organic solvents. Journal of Langmuir, 27(10), 6132-6137.
Castilon, J., Jones, B., & Kamo, K.) 2006.( Efficient regeneration of rose plants from somatic embryos of three genetically diverse cultivars. Floral Nursery Plants Research Unit, US National Arboretum, Beltsville, MD, USA.
Chandrasekaran, M., Lee, J. M., Moon, Ye. B., Jung, S. M., Kim, J., Kim, J. W., & Chul Chun, S. (2019). Isolation and characterization of a virulent and virulent strains of Agrobacterium tumefaciens from rose crown gall in selected regions of South Korea. Journal Plants, 8(11), 452. https:// doi: 10.3390/plants8110452.
Chung, H.J., Castro, C.M., Im, H., Lee, H., & Weissleder, R. (2013). A magneto-DNA nanoparticle system for rapid detection and phenotyping of bacteria. Journal of Nature Nanotechnology, 8(5), 369-375. https:// doi: 10.1038/nnano.2013.70.
Choi, O., Bae, J., Kang, B., Lee, Y., Kim, S., Fuqua, C., & Kim, J. (2019). Simple and economical biosensors for distinguishing Agrobacterium mediated plant galls from nematode-mediated root knots. The Journal of Scientific reports, 9, 17961. https://doi.org/10.1038/s41598-019-54568-2.
Izadi, Z., Sheikh-Zeinoddin, M., Ensafi, A.A., & Soleimanian-Zad, S. (2016). Fabrication of an electrochemical DNA-based biosensor for Bacillus cereus detection in milk and infant formula. Journal of Biosensors & Bioelectronics, 80, 582-589. https://doi: 10.1016/j.bios.2016.02.032.
Kafka, J., Pänke, O., Abendroth, B., & Lisdat. F. (2008). A label-free DNA sensor based on impedance spectroscopy. Journal of Electrochimica Acta, 53(25), 7467-7474. https:// doi: 10.1016/j.electacta.2008.01.031.
Khater, M., Escosura-Muñiz, A., & Merkoçi, A. (2017). Biosensors for plant pathogen detection. Journal of Biosensors & Bioelectronics, 93, 72-86. https:// doi: 10.1016/j.bios.2016.09.091.
Mafakheri, H., Taghavi, S. M., Puławska, J., Lajudie, P., Lassalle, F., & Osdaghi, E. (2019). Two novel genomospecies in the Agrobacterium tumefaciens Species.complex associated with rose crown gall. Journal of Phytopathology, 109, 1859-1868. https://doi.org/10.1094/PHYTO-05-19-0178-R
Mohd Said, N.A., Abu Bakar, N., & Lau, H.Y. (2018). Development of DNA biosensor for impedimetric detection of Erwinia Mallotivora (papaya dieback) as early warning system tool in plant disease management. National Conference on Agricultural and Food Mechanization (NCAFM). Conference April 2018. Mardi. Selangor.
Moattari, Gh., Izadi, Z., & Niaei, M. SH. (2021). Development of an electrochemical genosensor for detection of viral hemorrhagic septicemia virus (VHSV) using glycoprotein (G) gene probe, Aquaculture, 536, 736451. https://doi.org/10.1016/j.aquaculture.2021.736451.
Razdari, A.M., Ghasemi-Varnamkhasti, M., Izadi, Z., Rostami, S., Ensafi, A.A., Siadat, M., & Losson, E. (2019). Detection of sulfadimethoxine in meat samples using a novel electrochemical biosensor as a rapid analysis method. Journal of Food Composition and Analysis, 82,103252. https:// doi.org/10.1016/j.jfca.2019.103252.
Schaad, N.W., Jones, J.B., & Chun, W. (2001). Laboratory Guide for Identification of Plant Pathogenic Bacteria. Third Ed. USA: American Phytopathological Society, 373pp.
Wang, M., Gong, W., Meng, Q., & Zhang, Y. (2011). Electrochemical DNA impedance biosensor for the detection of DNA hybridization with polymeric film, single walled carbon nanotubes modified glassy carbon electrode. Russian Journal of Electrochemistry, 47(12), 1368-1373. https://doi.org/10.1134/S1023193511120123
Wei, J., Liu, H., Liu, F., Zhu, M., Zhou, X., & Xing, D. (2014). Miniaturized paper-based gene sensor for rapid and sensitive identification of contagious plant virus. Acs Applied Materials And Interfaces, 6(24), 22577-22584.Zhao, W., Lu, J., Ma, W., Xu, C., Kuang, H., & Zhu, S. )2011(. Rapid on-site detection of Acidovorax avenae subsp. citrulli by gold-labeled DNA strip sensor. Journal of Biosensors & Bioelectronics, 26(10), 4241-4244. https://doi: 10.1016/j.bios.2011.04.004.