Davis Pecan Variety Identification Using Specific DNA Fragment
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Solution Overview
Problem
Current methods for identifying Carya illinoensis varieties are inefficient due to reliance on morphological characteristics, which are prone to environmental and climatic variations, leading to unstable and inaccurate identification, and existing SSR marker methods are cumbersome and unreliable.
Innovation Solution
Development of a molecular specific labeled primer and a method using PCR amplification with specific primers to rapidly and accurately identify Carya illinoensis variety Davis, utilizing a characteristic sequence and a screening process involving reduced-representation sequencing and comparative analysis to design primers that produce a unique 359 bp DNA fragment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If morphological characteristics are used for identification, then the identification process is simple, but the accuracy and reliability are low due to environmental and climatic variations
Solution Approach 1:
The patent replaces the mechanical/visual inspection system (morphological observation) with a molecular biology system (PCR amplification and electrophoresis). By substituting direct visual assessment with DNA-based molecular markers, the method eliminates environmental influence while maintaining operational feasibility through standardized laboratory procedures.
Solution Approach 2:
The patent creates a molecular copy (DNA fingerprint) of the variety-specific genetic information. Instead of relying on variable phenotypic expressions, the method amplifies and analyzes specific DNA sequences that serve as stable copies of variety identity, independent of environmental conditions.
2Measurement precision
If SSR molecular marker methods are used, then the identification accuracy is improved, but the process becomes cumbersome and results become unstable
Solution Approach 1:
The patent extracts only the essential variety-specific DNA sequence information needed for identification, discarding the complexity of full SSR marker systems. By focusing on a single diagnostic DNA fragment with unique sequences for each variety, the method retains high accuracy while eliminating cumbersome procedures and instability issues.
Solution Approach 2:
The patent changes the identification parameter from variable phenotypic traits and complex SSR patterns to a simplified DNA sequence parameter. This parameter change enables the use of ordinary electrophoresis equipment instead of high-resolution sequencing, reducing device complexity while maintaining diagnostic accuracy.
3Measurement precision
If high-resolution sequencing is used for identification, then the measurement precision is improved, but the device complexity and cost increase
Solution Approach 1:
The patent applies partial action by amplifying and sequencing only the specific diagnostic DNA fragment containing variety-specific information, rather than performing whole-genome or full-gene sequencing. This partial sequencing approach achieves sufficient identification precision using ordinary electrophoresis equipment, avoiding the need for complex high-resolution sequencing devices.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method provides a simple, rapid, and accurate means of identifying Carya illinoensis variety Davis, with high reliability and specificity, allowing for precise differentiation from other varieties using ordinary electrophoresis, reducing the need for high-resolution sequencing and sample complexity.
Implementation Method 1
performing PCR amplification using the genomic DNA provided in the step 1) as a template and the molecular specific labeled primer as an amplification primer
Implementation Method 2
carrying out electrophoresis detection of the amplified product of the step 2)
Data Source
AI summary
The present invention discloses a highly specific characteristic sequence, molecular specific labeled primer of Carya illinoensis variety Davis and applications thereof, and further discloses a method for rapidly identifying Carya illinoensis variety Davis. Wherein, the sequence of the molecular specific labeled primer is: upstream primer 5′-TCCTGAAAGCAGCCACAACA-3′ (SEQ ID NO:3); downstream primer 5′-GACATGTGTACGAGGTGGTCA-3′ (SEQ ID NO:4). The molecular specific labeled primer of the present invention can be used for early identification of the Carya illinoensis variety Davis rapidly and easily distinguishing it from other Carya illinoensis varieties, providing a strong technical support for the identification of Davis and planting resource protection. The method for rapid identification of the Carya illinoensis variety Davis is simple, rapid and accurate, and it is an irreplaceable molecular means for identifying Carya illinoensis variety by apparent characteristics.