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Method of detecting crop seed purity

A technology for crops and seeds, applied in the direction of biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc., can solve the problems of complex seed purity experiment technology, high experimental condition requirements, and promotion restrictions, so as to improve sensitivity, The effect of strong polymorphism and low cost

Inactive Publication Date: 2005-06-22
TIANJIN NORMAL UNIVERSITY
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Problems solved by technology

This method has positive significance in the identification of true and false seeds of a certain crop, but it also has the characteristics of complicated experimental technology, high experimental conditions, and high cost when used in the detection of seed purity, so its promotion in production is subject to certain restrictions. limit

Method used

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  • Method of detecting crop seed purity
  • Method of detecting crop seed purity
  • Method of detecting crop seed purity

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Disinfect the cucumber seeds with 10% Antiformin for 10 minutes, rinse thoroughly with distilled water, soak the seeds at 27°C for 24 hours, transfer them to a petri dish, and culture them at (24±1)°C in the dark for 2-3 days, and take the yellowed seeds. Seedling hypocotyls were used as experimental materials. Protein sample preparation and SDS-PAGE trace protein electrophoresis can identify the authenticity and purity of crop seeds by analyzing the possible differences in the protein fingerprints of individual seedlings. Specifically, take 0.1g of the fresh and tender tissue of crop seed seedlings, that is, the experimental material, and cut it into pieces , add 1ml of 0.1M / L pH7.0 phosphate buffer, after homogenization, put it in a refrigerated centrifuge and centrifuge at 5000r / min for 5min, take 0.2ml of the supernatant, add an equal volume of protein denaturation solution, denature in a boiling water bath for 3min, cool down, Take 10ul for microprotein SDS-PAGE an...

Embodiment 2

[0060] The kidney bean seeds are processed, the method is the same as in Example 1, and the kidney bean hypocotyl is selected as the experimental material. Others are with embodiment 1. figure 2 Shows the microprotein SDS-PAGE silver-stained fingerprints of hypocotyls of kidney bean. Kidney bean seedlings are rich in protein, and at least 50 bands have been detected in hypocotyls, and the number of bands, spectrum type, and band coloration among individuals are completely consistent, indicating that the seeds are of high purity.

Embodiment 3

[0061] Embodiment 3: the kidney bean seed is processed, and its method is the same as embodiment 1, and the kidney bean radicle is selected as the experimental material. Others are with embodiment 1. image 3 The microprotein SDS-PAGE silver-stained fingerprints of kidney bean radicle are shown. Kidney bean seedlings are relatively rich in protein, and the radicle contains more than 60 bands. The number of bands, spectrum type, and band coloration among individuals are completely consistent, indicating that the seeds are of high purity.

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Abstract

A method for detecting the purity of crop seeds relates to a method for detecting the purity of crop seeds by SDS-PAGE trace protein electrophoresis combined with ultra-sensitive silver staining. The invention solves the problem of labor-intensive and time-consuming detection of traditional field planting, and the detection result is easily affected by the environment; the RAPD technical method has complicated experimental techniques, high requirements for experimental conditions, and high cost; there are few bands of isozymes and stored proteins, the method is complicated, and the technology is difficult Large and costly problems. The method utilizes the difference of protein types contained in crop seeds, and uses micro-protein electrophoresis and ultra-sensitive silver staining techniques to identify the purity of the seeds. The method comprises the steps of crop seed treatment, SDS-PAGE micro protein electrophoresis, ultrasensitive silver staining and result analysis. Compared with the existing methods, the detection method of the present invention has the advantages of simplicity, high sensitivity, many protein bands, high efficiency, rapidity, low cost and easy popularization and application.

Description

technical field [0001] The invention belongs to a method for detecting the purity of crop seeds, in particular to a method for detecting the purity of crop seeds by SDS-PAGE trace protein electrophoresis combined with supersensitive silver staining. technical background [0002] The authenticity and purity detection of seeds is an important task in seed production, which is directly related to the development of agriculture and the vital interests of farmers, so it is highly valued by the state. The traditional method for seed companies to test the authenticity and purity of seeds is field planting, that is, growth testing, which is to sow representative seeds, and judge the authenticity and purity by the phenotype of the adult plants. However, this detection method is labor-intensive and time-consuming, and the detection results are easily affected by the environment. [0003] At present, protein fingerprinting technology has high resolution, strong polymorphism, reproduci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N27/447
Inventor 陈宏程罗根王振英彭永康
Owner TIANJIN NORMAL UNIVERSITY
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