Single-grain prolamin extraction method that can meet the requirements for purity identification of millet hybrids

Abstract

The invention discloses a single-grain prolamin extraction method that can meet the purity identification requirements of millet hybrids. The prolamin extract used has the following components: 10 parts by volume of NN-dimethylformamide, and 20 parts by weight of sucrose parts, 50 parts by volume of isopropanol, and deionized water to supplement the extraction solution to 100 parts by volume; and during the prolamin extraction process, the extraction temperature was controlled to be 65°C. The method of the invention establishes a set of prolamin extraction methods suitable for millet grains, and provides strong technical support for carrying out the purity identification, variety management and large-scale promotion research of millet hybrids.

Description

technical field [0001] The invention relates to a plant prolamin extraction technology, in particular to a method for extracting prolamin from a single millet seed. Background technique [0002] Before the 1990s, the identification of millet hybrids was mainly based on the difference in the seedling color of the parental parent. Dominant inheritance law, the hybrid is the same color as the male parent, by pulling out the female parent with the lighter color mixed in the hybrid, removing the hybrid, and also by investigating the proportion of the sterile plant with the lighter color to determine the purity of the hybrid Although this method can remove some hybrids in hybrids, it limits the application of sterile lines with darker colors, is labor-intensive and time-consuming, cannot remove hybrid restorers and other materials mixed in, and the purity of identification is not precise. After the 1990s, with the successful introduction and creation of herbicide-resistant mater...

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Problems solved by technology

[0002] Before the 1990s, the identification of millet hybrids was mainly based on the difference in the seedling color of the parents. Conventional materials with dark seedling color were selected as the male parent, and the male sterile line with lighter seedling color was used as the female parent. Dominant genetic law, the color of the hybrid is the same as that of the male parent, the hybrid is removed by removing the female parent with a lighter color mixed in the hybrid, and the purity of the hybrid is determined by investigating the proportion of the sterile plants with a lighter color Although this method can remove some hybrids in hybrids, it limits the application of darker sterile lines, and is labor-intensive and time-consuming. It cannot remove mixed restorer lines and other materials in hybrids, and the purity of identification is not high. precise

After the 1990s, with the introduction and successful creation of herbicide-resistant materials, through the cultivation of herbicide-resistant restorer lines, the sterile lines are not resistant to herbicides, and the characteristics of herbicide-resistant traits are dominantly inherited, through the seedling stage. Spraying herbicides to kill the false hybrids is used to identify the purity of the hybrids and remove the false hybrids. This method enables the application of CMS lines of various seedling colors, but cannot identify the restorer lines mixed with hybrids. or other herbicide-resistant materials, still cannot completely remove the false hybrids in the hybrids, and cannot accurately identify the purity of the hybrids

[0006] Through research, some millet researchers have concluded that salt-soluble protein can be used for variety identification [13] , but further studies have shown that most of the non-prolamins such as salt-soluble proteins are non-storage proteins that participate in the immune response, and their polymorphism is poor, which is not suitable for the identification of variety purity. [14] , on this basis, the researchers began to try to use the wheat prolamin A-PAGE method to identify polymorphisms of glutinous prolamin [5,14,15,16] , but due to the small grain size of millet, more than 10 seeds are needed to extract prolamin from millet [14] , and the variety purity identification must be based on each seed, therefore, this method cannot be used for the purity identification of millet hybrids; researchers have begun to use the SDS-PAGE method to identify polymorphisms of millet prolamins [17] However, due to the shortcoming of long electrophoresis time in SDS-PAGE, and when the concentration of isopropanol is increased to 50%, more than 20 grains of prolamin are needed to extract prolamin and detect polymorphism , can not meet the needs of the identification of the purity of millet hybrids

So far, the prolamin polymorphisms of glutinous rice cannot be applied to the identification of genetic diversity and variety purity

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