MARGDARSHAN SANSTHAN

AGRICULTURE COLLEGE AMBIKAPUR ,

SURGUJA(C.G.)

TOPIC – DETECTION OF GENETICALLY

MODIFIED CROPS
Course Name– Principle of Seed Technology
Course Code– APB-5221

SUBMITTED TO:- SUBMITTED By:-

ASSISTANT PROFESSER BHAVANA PRADHAN
ANJANI KUMAR JHA B.Sc. (Ag.) 2nd year 2nd sem.
INDEX
 GM/ Transgenic crops
 Streams where GM detection is required
 Levels of detection
 Detection method
 Protein based method
 DNA based method
GM /Transgenic Crops
 Characterized by the insertion of new
genes or sets of genes into their genome.
 The new genes translate and new
proteins exposed
 This gives the plant new characteristics.
Example of transgenic plants

 Bt cotton-
Cotton plants resistant to lepidopteran
insects.
 Round up ready soybean –
Soybean resistant to glyphosate
 Golden rice-
Rice grains with beta carotene
GM Detection is Required by
 GM Developer (Institutions and Industry)
• To assure purity and segregation of different
events
• To trace the genetic modifications
 Seed companies, Food & feed industry
• To assure purity and segregation of products
• To comply with the regulations
 Competent (enforcement) authorities
• To ensure compliance with legislation
• To solve the legal dispute if they arise
LEVELS OF DETECTION
 • Screening: to determine whether a product is
GM or not. For this purpose, a general screening
method can be used. The result is a
positive/negative statement.
 • Identification: to find out which GM crop or
product are present and whether they are
authorized or not in the country.
 • Quantification: If a crop or its product has been
shown to contain GM varieties, then it become
necessary to assess compliance with the
threshold Regulation by the determination of the
amount of each of the GM variety present.
Detection Methods
• Basis of detection is to exploit the
differences between the unmodified
variety and the transgenic plants , which
are new genes and proteins expressed by
them.
 • Accordingly testing methods look for –
the genes (DNA) introduced into the
crop – the proteins produced in the plant
by the introduced gene
Detection methods – requirements
• Target molecules
• DNA or protein
• Capture molecules
• For DNA: primers and probes
• For proteins: antibodies
• Reference materials
• Positive and negative controls
• Calibrants for quantitation
Protein based methods
 • Lateral flow strip
 • ELISA
Lateral flow strips
 • Collect leaf/seed and extract sample
 • Place the strip into the extraction tube
 • The sample will travel up the strip
 • Allow the strip to develop for 10
minutes
 • To retain the strip, cut off and • Discard
bottom section of the strip
Enzyme Linked Immunosorbent
Assay (ELISA)
 1. Prepare a surface to which a known quantity of
antibody is bound.
 2. Apply the antigen-containing sample to the
plate. Wash the plate, so that unbound antigen is
removed.
 3. Apply the enzyme-linked antibodies which are
also specific to the antigen. Wash the plate, so
that unbound enzyme-linked antibodies are
removed.
 4. Apply a chemical which is converted by the
enzyme into a fluorescent signal.
 5.View the result: if it fluoresces, then the sample
contained antigen
Protein based methods: Advantages
 • Relatively cheap to perform
 • Rapid turnaround - 5 to 20 mins for strips;
24 hours for ELISA
 • Strips do not require trained personnel
 • Reasonably sensitive
 • Less susceptible to ‘false positives’
 • Low per sample cost
 • Handles large number of samples
 • Can be subjected to automation
 • Detection kits available commercially
Protein based methods :
Disadvantages
 • Not appropriate for some GM varieties
when the GM protein is only produced in
the leaves or stems and not in the actual
grain. Protein tests on the grain are
therefore not informative.
 • In processed foods the proteins denatures
easily, which makes it difficult to use ELISA
for processed food fractions.
 • Limited to one or a small number of
protein per test
 • Not very sensitive (~1% of GM protein)
DNA based methods
 • DNA based methods are based on
detection of the specific genes, or DNA
genetically engineered into the crop.
 • Commercial testing is conducted using
PCR technology
 • The PCR technique is based on
multiplying a specific target DNA allowing
the million or billion fold amplification by
two synthetic oligonucleotide primers.
DNA based Method
PCR based detection
 Detects a known DNA sequence by
amplification
 • Target sequence is duplicated billions of
times
 • Results obtained can be quantitative or
qualitative
 • Enzyme :Taq polymerase • Equipment :
Thermocycler/PCR machine
GM ANALYSIS FLOW CHART
 STANDARD REFERENCE (+ve Contro)
 NON GM FOOD (-ve Control)
 GM FOOD MATERIAL

DNA Purity

PCR Amplification

DNA Electrophoresis
Interpretation & Documentation
DNA based methods: Advantages
 High sensitivity. Detection limits ≤ 0.1% of
DNA present
 • DNA being highly stable , these methods
work with most product types - both
processed and unprocessed products
DNA based methods :
Disadvantages
 Require relatively advanced laboratory
facilities and instrumentation and highly
trained staff
 Time (usually >24 hrs for results)
 Special precautions need to be taken to
avoid the contamination
 More expensive than Protein Based
Method
REFERENCE
https://www.agrilearn.tel
THANK YOU