Document

‫التفكير العلمي وأعداد التقارير العلميه‬
‫(زرع ‪) 201‬‬
‫تلخيص بحث رقم(‪)1‬‬
‫عنوان البحث‪ /‬المحاصيل المعدله‬

‫وراثيآ‬
‫االسم ‪ :‬چودي تامر صالح الدين‬
‫الرقم الموحد‪23231689 :‬‬
) Genetically modified corops )
1: Jinbin Wanga,b,1,
Yu Wanga,c,1
2: , Xiuwen Hua,
Yifan Chena,b
3:, Wei Jianga,b, Xiaofeng Liuc, Juan Liud,e,
Lemei Zhud,e, Haijuan Zenga,b,
4:*, Hua Liua,b,*
Joudy Tamer Salah

1-Introduction
• In recent years, the wide-scale application of genetically modified crops has been beneficial for
the development of environmentally friendly agricultural products[1]. However, disputes
regarding genetically modified (GM) crops have been persistent[2], and corresponding risk
assessment and safety testing efforts are necessary and urgently required[3,4]. In 2019, the
number of stacked insect resistance/herbicide tolerance (IR/HT) traits grew by 6%, and these
crops covered 45% of the global GM crop acreage, making stacked IR/HT traits the dominant
trait in GM crops for the first time[5]. Thus, related IR/HT genes have become the mainstream
target genes for rapid transgenic testing. CP4-EPSPS, pat, and bar are the genes commonly found
in herbicide-tolerant crops, and most commercial glyphosate-resistant crops contain CP4-EPSPS
genes[6]. Additionally, the Cry gene of Bacillus thuringiensis (Bt), which is able to resist a wide
range of pests (Lepidopterons, Coleopterans, and Dipterans), has been widely employed in most
insect-resistant transgenic crops[7].
• Many nucleic acid or protein-based transgenic detection methods have been established and
commercially applied to monitor GM crops. DNA-based polymerase chain reaction (PCR) and
protein-based
enzyme-linked immunosorbent assays (ELISA) are often regarded as the traditional methods for GM
crops detection[8,9]. Protein-based immunological methods used to be popular for in situ detection due
to their simplicity, speed and ease of operation, but these methods are based on active structures and
thus susceptible to poor interference resistance and sample denaturation[10]. PCR methodologies rely
on a variety of instruments and equipment, and have high requirements for skilled operators and
laboratory conditions[11]. Although these methods (PCR and ELISA) have excellent reliability and
stability in practical applications, with the increasing number of GM crops yield and complex stacked
traits, more advanced techniques are needed to enable efficient detection in outdoor environments.
Currently, many new techniques for nucleic acid amplification have circumvented the limitations of
traditional PCR methods in terms of time and equipment, providing new tools for GM crops
detection[12], such as loop-mediated isothermal amplification (LAMP), helicase-dependent amplification
(HDA), and recombinase polymerase amplification (RPA)[13,14]. Among them, RPA has been widely used
for nucleic acid detection because of its high efficiency and easily achievable reaction temperature
(37−42 °C)[15]. Recently, strategies such as RPA combined with agarose gel electrophoresis (RPA-
AGE)[16], real-time fluorescent RPA (RT-RPA)[15,17], RPA combined with DNA fluorescent dye (RPA-SYBR
Green I)[18,19], RPA combined with lateral flow immunochromatography (RPA-LFD)[20,21] and RPA
combined with Cas12a cleavage system (RPA-Cas12a-FS)[22] have been established and implemented in
the field of rapid detection. Among these methods, the test strip method is the most suitable for on-site
testing for its full independence of testing equipment.
In this study, a dual RPA system was constructed by labeling the forward and reverse fragments of the
two primer pairs separately, aiming to achieve simultaneous detection of CP4-EPSPS and Bt-Cry1Ab/Ac
genes using a single immunochromatographic test strip. For this strategy, the amplification products of
two different markers in one reaction system can be captured by the corresponding antibodies on the
test strips respectively, and the corresponding antibody-fixed zone turns red, which can be judged by the
naked eye. Without relying on equipment such as a PCR instrument or an electrophoresis instrument,
this procedure can be performed in a field environment and meet the needs of rapid detection for
simultaneous identification of CP4-EPSPS and Bt-Cry1Ab/Ac genes. Besides, the results show that the
method possesses good detection performance
2: Materials and methods

• Ultra-Rapid Nucleic Acid Release Agents (for DNA)
were purchased from Weifang Ample Future
Biotechnology (Weifang, China). TwistAmp® Basic
Kits were purchased from Morey Biosciences
(Shanghai, China). The oligonucleotides needed were
synthesized and purified by Sangon Biotech
(Shanghai, China). r-SA, Digoxin antibody of LSBio,
goat anti-rabbit immunoglobulin G (IgG) antibody,
and colloidal gold-labeled fluorescein isothiocyanate
(FITC) antibody were from Beijing Bersee Science and
Technology Co., Ltd. (Beijing, China), Beijing
Biocreative Technology Co., Ltd.
3: Results and Discussion
References :
[1] A. Seid, B. Andualem, The role of green

biotechnology through genetic engineering for climate
change mitigation and adaptation, and for food security:
current challenges and future perspectives, J. Adv. Biol.
Biotechnol. 24 (2021) 1-11.
https://doi.org/10.9734/jabb/2021/v24i130192.
[2] B. Martinez-Ribaya, F.J. Areal, Is there an opportunity
for product differentiation between GM and non-GM
soya-based products in Argentina?, Food Control. 109
(2020) 106895.
[3] M.P. Pimbert, B. Barry, Let the people decide: citizen
deliberation on the role of GMOs in Mali’s agriculture,
Agr. Human Values. 38 (2021) 1097- 1122.
https://doi.org/10.1007/s10460-021-10221-1.
[4] M.F. Eckerstorfer, M. Grabowski, M. Lener, et al.,
Biosafety of genome editing

Document

Uploaded by

Copyright:

Available Formats

Document

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Document

Uploaded by

Copyright:

Available Formats

‫التفكير العلمي وأعداد التقارير العلميه‬

‫عنوان البحث‪ /‬المحاصيل المعدله‬

Joudy Tamer Salah

2: Materials and methods

[1] A. Seid, B. Andualem, The role of green

You might also like