An image analysis for determination of genetic purity of different wheat varieties
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Transcript of An image analysis for determination of genetic purity of different wheat varieties
E. Nasrollahi, G. Tohidloo, S. Mobasser, M. Dehghanshoar, F. Khazaei, M. [email protected]
An image analysis for determination of genetic purity of different wheat varieties
First: A view over the prevalent methods Second: Introducing a new method for genetic purity determination Finally: considering validation test of the new method
Seed quality factors are as follows:
Improved varieties
Genetic purity
Physical purity
Seed viability
Seed vigor
Seed health
Seed moisture
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Genetic PurityGenetic purity refers to the percentage of contamination by seeds or genetic material of other varieties or species.
seedquest.com4
Prevalent methods for determination of genetic purity are as follows: 1. Control plot method2. Phenol test 3. Molecular method
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Off-type
1 .Control plot method
Disadvantages Advantages
High cost Relatively precise
Time consuming No special equipment
Labor force
Expert
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Disadvantages Advantages
Low precision Rapid
Expert Low cost
More human errors No special equipment
2. Phenol test
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3 .Molecular method
Disadvantages Advantages
High cost Rapid
Complicated and difficult Precise More experience
Advanced equipment
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We are looking for a technique to:• Increase test precision• Decrease the costs • Reduce human errors• And save time
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Biochemical and Imaging Technique for Analysis (BITA)This technique is developed by the present research for determination of genetic purity
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2- Washing and Soaking the seeds The seeds were washed and soaked in distilled water and kept at room temperature for 20 h.
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3- Seed replacement and adding Phenol Each seed was placed in 0.5mL tube. Then, 400 µL of 1% phenol solution was added to the tube and kept at room temperature
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4- Sampling of phenol solution 250µL of solution was added to Elisa testwells and aniline blue solution was used as index points.
index points19
5- Image capturing of the platesThe plates were placed on Hp Scanner (G4050) and images were captured.
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6- Image processing:6-1- Automatically Rotating of the imagesThe images were rotated automatically and kept in correct position using Pattern of index points.
index point
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6-2- Identification of test wells center: Identification of the center of all test wells and making a circle with 200 pixel area for each center of test well
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6-3- Converting RGB to HSV mode:RGB color mode converted to HSV mode and the colors of pixels were averaged.
(RGB)RED, GREEN and BLUE
(HSV)HUE, SATURATION and VALUE
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Colors classification
classification of colors and
determination of their percentage
EXAMPLE FIGURE
7 -Image Analysis
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A B
A+B
AB1A=10%B=90%
AB2 A=15%B=85%
AB3 A=20%B=80%
R4R3R2R1R4R3R2R1R4R3R2R1
Validation of BITAValidation of the software and methodExperimental material: two samples of nucleus seed class of wheat were mixed in 10, 15 and 20% each with 4 replications
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AB110% impurity
AB215% impurity
AB320% impurity
R1 R2 R3 R4
R1 R2 R3 R4
R1 R2 R3 R4
Software input
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AB1/R4
This color shows impurity of a variety
Off types were also distinguished and recorded
Variety A
Variety B
unknown variety
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Disadvantages Advantages
Required Software Rapid(It takes 3 days)
Precise
Simplifies the performance and easy to use
Low cost
No special equipment
Low human errors
keeping data and retrieval of the test information
BITA-technique
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Rapid Precise Simplifies the performance
Easy to use cost special
equipmenthuman errors
keeping data
Required Software
Control plot method o
Phenol test
Molecular method o o
BITA + + + + + + + +
+ + +++++++
+ + +
+ :Positive point - : negative point O : Relative point
- - - -- -
--
----
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AdvantagesMethods
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• BITA software and method can be used for determination of genetic purity in wheat varieties
• The method has more advantages in comparison with prevalent methods
• BITA Software can keep the data and retrieval of the test information
• Despite the efficiency of this method, it should be validate by ISTA laboratories as well
Conclusion
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