Volume 9, Issue 2 (2018)
JMBS 2018, 9(2): 213-218 |
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Karimi F, Khodaie E. Efficiency of PCR and Nanobiosensor Methods for Detection of Transplastomic Tobacco Plant. JMBS 2018; 9 (2) :213-218
URL: http://biot.modares.ac.ir/article-22-24442-en.html
URL: http://biot.modares.ac.ir/article-22-24442-en.html
1- Biology Department, Sciences Faculty, Maragheh University, Maraghrh, Iran, Maragheh University, Daneshgah Street, Madar Square, Amir Kabir Highway, Maraghrh, Iran. Postal Code: 55181883111 , karimifm@maragheh.ac.ir
2- Biotechnology Department, Agriculture Faculty, Maragheh Branch, Islamic Azad University, Maragheh, Iran
2- Biotechnology Department, Agriculture Faculty, Maragheh Branch, Islamic Azad University, Maragheh, Iran
Abstract: (6896 Views)
Aims: In recent years, according the benefits of chloroplast transformation, the cultivation of transplastomic plants and their products have been increased. Due to their biosafety concerns, their identification and labeling have become more widely considered. The aim of this study was to present an optimal method based on polymerase chain reaction (PCR) and nanobiosensor for detection of transplastomic tobacco plants and compare their sensitivity.
Materials and Methods: In the present experimental research, aadA gene as a chloroplast selectable marker was considered to design specific primer and probe. In PCR method, after optimization of aadA gene amplification, its sensitivity was evaluated with different percentages of transplastomic DNA. In nanobiosensor method at first, the labeled aadA probe was immobilized on graphene oxide (GO) and, then, hybridization reaction was optimized to identify target DNA sequence.
Findings: The amplification of 800 bp DNA related to aadA gene was observed. The PCR reaction allowed up to 5% DNA transplostomy tobacco to reproduce the aadA gene. In results of nanobiosensor after immobilization of aadA probe on GO, fluorescence emission was quenched and by adding the trasplastomic tobacco, DNA was observed again. In this method, up to 1% transplastomic tobacco DNA, fluorescence emission was significant in comparison with control tobacco plant.
Conclusion: The PCR method can detect a transplastomic tobacco plant with 5% DNA sensitivity and detect biomarker sensitivity with 1% DNA sensitivity.
The PCR method can detect a transplastomic tobacco plant with 5% DNA sensitivity and nanobiosensor can detect with 1% DNA sensitivity. Therefore, nanobiosensor method is not only a reliable diagnostic method, in addition to the PCR method for detecting transplastomic plants, but also has a higher sensitivity.
Materials and Methods: In the present experimental research, aadA gene as a chloroplast selectable marker was considered to design specific primer and probe. In PCR method, after optimization of aadA gene amplification, its sensitivity was evaluated with different percentages of transplastomic DNA. In nanobiosensor method at first, the labeled aadA probe was immobilized on graphene oxide (GO) and, then, hybridization reaction was optimized to identify target DNA sequence.
Findings: The amplification of 800 bp DNA related to aadA gene was observed. The PCR reaction allowed up to 5% DNA transplostomy tobacco to reproduce the aadA gene. In results of nanobiosensor after immobilization of aadA probe on GO, fluorescence emission was quenched and by adding the trasplastomic tobacco, DNA was observed again. In this method, up to 1% transplastomic tobacco DNA, fluorescence emission was significant in comparison with control tobacco plant.
Conclusion: The PCR method can detect a transplastomic tobacco plant with 5% DNA sensitivity and detect biomarker sensitivity with 1% DNA sensitivity.
The PCR method can detect a transplastomic tobacco plant with 5% DNA sensitivity and nanobiosensor can detect with 1% DNA sensitivity. Therefore, nanobiosensor method is not only a reliable diagnostic method, in addition to the PCR method for detecting transplastomic plants, but also has a higher sensitivity.
Subject:
Agricultural Biotechnology
Received: 2018/08/25 | Accepted: 2018/08/25 | Published: 2018/10/2
Received: 2018/08/25 | Accepted: 2018/08/25 | Published: 2018/10/2
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