Method for identifying wheat variety using wheat water-solubility protein by high-efficiency capillary electrophoresis

Abstract

The invention relates to a method to quickly verify the varieties of wheat, which comprises sampling; sample preparation; capillary electrophoresis; data processing analysis; and establishing standard high efficiency capillary electropherograms of wheat protoproteose to verify the types or idioplasm of wheat. The invention is characterized in that the end of a single seed without an embryo weighing about 15 to 20mg is taken, and is added to the ethanol of 20 percent in the way that 10 microlitres are added to every 1mg of the ethanol; the mixture is shaken for fifteen minutes in the room temperature, and is centrifugated at the speed of 10000rpm for ten minutes. The supernatant is the capillary electrophoresis specimen; the electrophoretic buffer during high efficiency capillary electrophoresis is phosphoric acid and glycine buffer solution of 0.1mol and the PH value is 2.5. Acetonitrile of 20 percent and hydroxyprolyl-methyl cellulose of 0.05 percent are added in the buffer solution.In addition, concrete electrophoretic conditions and parameters and flushing procedures are given. Compared with the prior method to verify the varieties of wheat, the method has the characteristics of quickness, accurateness, low cost, and high repeatability.

Description

technical field [0001] The invention relates to a rapid identification method of wheat varieties, in particular to rapidly separating wheat water-soluble protein by means of high-efficiency capillary electrophoresis and constructing a standard map, and then realizing the identification method of wheat varieties on this basis. Background technique [0002] Wheat is one of the most important food crops in the world and an important protein source in human food consumption. my country has always been a major producer and consumer of wheat in the world, with the sown area and total output ranking first in the world. As an important part of wheat seed protein, water-soluble proteins have received more and more attention. In wheat seeds, water-soluble proteins include albumin and globulin, mainly enzymes in some metabolic processes , inhibitory factors, etc., such as α-amylase, protease inhibitors, regulatory enzymes, synthetases of special pathways, metabolic enzymes, etc., the ...

AI Technical Summary

Problems solved by technology

However, the above-mentioned methods have limitations to varying degrees, such as morphological characteristics are easily affected by environmental factors, and the reliability is poor; isoenzyme and protein electrophoresis separation time is long, labor-intensive and time-consuming, and in most cases it takes 1-2 days to obtain the analysis results , and the resolution is low, it is often impossible to distinguish some closely related species, and its resolution and repeatability are also poor, making it difficult to carry out quantitative analysis and comparison between the results obtained by different laboratories

In addition, the PAGE method must use neurotoxic acrylamide and other contaminating reagents; although RP-HPLC has a high degree of automation, the equipment is expensive, the analysis cost is high, and the resolution is difficult to meet the needs; DNA molecular labeling techniques, such as PCR, RAPD , AFLP, etc. In recent years, there have been many studies and high accuracy, but the technology is complicated, labor-intensive and time-consuming, and the cost of identification is high, so it is difficult to be widely used

Therefore, there are some shortcomings in the existing wheat variety identification methods.

Images