Fingerprinting of crop varieties using DNA markers
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Transcript of Fingerprinting of crop varieties using DNA markers
7/30/2019 Fingerprinting of crop varieties using DNA markers
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Lecture 32
Fingerprinting of crop varieties using DNA markers
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DNA fingerprinting
Breeder Seed
+ Control
Seed Lots
Test sample
DNA extraction
DNA Purity
DNA amplification
DNA electrophoresis
Interpretation &
Documentation
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Markers used for Fingerprinting
• RAPD
• RFLP
• SSR
• ISSR
• AFLP
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Repetitive sequence
primer I
primer II
plant A
plant B
microsatellite
plant A
plant B
flanking region IIflanking region I
specific primers were designed corresponding to
flanking sequence of microsatellite
PCR analysis and analyze on 6%denaturing
polyacrylamide gel with silver staining
A B
Schematic of SSR assay
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Gel electrophoresis
Protocol
•
Gel electrophoresis is a technique used to separate DNA samples based on their size.• DNA is loaded onto an agarose gel which is placed in a buffer solution.
• An electric current will cause the DNA (which is positively charged) to migrate to the negativecurrent (down the gel).
• The small DNA molecules will move faster, and this results in small DNA fragments movingfarther down the gel.
7/30/2019 Fingerprinting of crop varieties using DNA markers
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Gel electrophoresis
Ladder
Breeder SeedSample Seed
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Seed Purity Assessment Of Rice Hybrid Using
Microsatellite Markers
Detection of impurities in the Indian rice hybrid-KRH2Through multiplex PCR using the microsatellite markers RM164 and
RM206. M — 50 bp ladder, A — CMS line (IR58025A), H — Hybrid
(KRH2), R — Restorer line (KMR3), 221 to 240 — Samples of hybrid
KRH2 collected from a commercial seed-lot.
Arrow indicates contaminants
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DNA Fingerprint of Maize genotypes
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Problems & Precautions
• Marker identification
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SuitablePrimer !!!
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Advantages of DNA fingerprinting in plants
- identification of genetic diversity within a breeding population.
- RFLP markers are used to detect the genetic distance in wheat.
-RAPD markers are used for characterization, estimation of genetic relatedness
and determination of genetic diversity of tea germplasm.
- to find genetic relatedness and difference in figs.
- AFLP markers help in assessing genetic diversity among cultivars such as
wheat. It also helps detect higher level of polymorphism.
- DNA fingerprinting of herbal drugs useful in authenticating the various claims
of medical uses related to the plants.
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Simplifying the Search
DNA fingerprinting can be of use to plant breeders to simplify their work and reduce
the amount of time it takes to produce crops with desirable new traits.
Guaranteeing Crops
Protecting plant breeders's rights (the breeders' patents on specific types of seed), is
another use for the DNA test. Disputes over the true identify of seed varieties can be
easily resolved, since the test will be able to isolate the specific traits that distinguishone seed variety from another
Building a Library
In order for the seed industry or others to effectively use DNA tests, both private and
government labs are working on building a library of crop DNA profiles. As newsamples are analyzed, a computer scan can produce matches between the samples
and DNA profiles already recorded. These labs plan to offer the testing service at a
reasonable cost. That means DNA testing will likely becomes as commonplace as
other agricultural testing services, like soil sampling and seed germination testing.