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Method for identifying purity of varieties or germplasm of conventional varieties of wine making glutinous sorghum

A technology for sorghum and varieties, applied in the field of molecular biology, can solve the problems of narrow genetic basis and high similarity, and achieve the effects of good repeatability, strong specificity and wide identification range

Active Publication Date: 2017-10-17
KWEICHOW MOUTAI COMPANY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the genetic basis of conventional sorghum species in my country is relatively narrow, and the similarity between some genomic loci of conventional sorghum species represented by Hongyingzi is very high. Most of the amplified SSR molecular marker loci have similar band sizes. Therefore, how to screen out Identifying the specific SSR markers corresponding to the varieties is a key point of the present invention, and it is also a difficult point
At present, there is no report on the identification method of glutinous sorghum germplasm molecular marker technology for wine-making sorghum, especially Maotai-flavored wine

Method used

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  • Method for identifying purity of varieties or germplasm of conventional varieties of wine making glutinous sorghum
  • Method for identifying purity of varieties or germplasm of conventional varieties of wine making glutinous sorghum
  • Method for identifying purity of varieties or germplasm of conventional varieties of wine making glutinous sorghum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Screening of SSR molecular marker sites and primers

[0056] Experimental steps:

[0057] Step 1. According to Gramene database (http: / / archive.gramene.org / cmap / ) published sorghum SSR molecular marker information to synthesize primers, it is required to randomly select SSR sites and primers on the 10 chromosomes of sorghum for synthesis. The synthesized primers are shown in Table 1;

[0058] Step 2. Collect the main wine-growing sorghum varieties in the country, and extract the genomic DNA of sorghum varieties;

[0059] Step 3. Screening of variety-specific primers. Use the synthesized primers (Table 1) to perform PCR (Polymerase Chain Reaction) amplification and electrophoresis analysis on the genomic DNA of different sorghum varieties, and finally screen out the sorghum varieties that can be used for winemaking. Specific SSR molecular markers identified;

[0060] Among them, the specific extraction of sorghum genomic DNA is:

[0061] Extract Hongyingzi, Qiangao 7, Q...

Embodiment 2

[0086] Example 2 Identification of variety / germplasm purity in different mixed samples of sorghum

[0087] In order to illustrate the amplification of a single molecular marker in two mixed samples of sorghum, the inventors mixed different sorghum varieties in pairs and then accelerated the germination, then extracted DNA, amplified with marker txp287 and analyzed by gel electrophoresis, the results are as follows Figure 13 Shown.

[0088] Table 2

[0089]

[0090] Numbers 1-5 are two sorghum varieties mixed at a ratio of 1:1, and numbers 6-14 are two sorghum varieties mixed at a ratio of 4:1.

[0091] Taking the red tassel sorghum as an example, the results of mixed samples of No. 6 and 7 containing red tassel sorghum varieties showed that even if a small amount of components were present, obvious difference bands could be amplified. Among them, Hongyingzi and Liaoza 37 can only amplify bands with a size of 250 bp at the txp287 labeling site, while control 1 can amplify bands with a...

Embodiment 3

[0092] Example 3 Identification of variety / germplasm purity in different mixed samples of sorghum

[0093] In order to illustrate the amplification of a single molecular marker in different sorghum mixed samples, the inventors mixed two or three different sorghum varieties and then accelerated the germination, then extracted DNA, amplified it with txp36 and analyzed it by gel electrophoresis. The results are as follows Figure 14 Shown.

[0094] table 3

[0095]

[0096] Numbers 1-4 are two sorghum varieties mixed at a ratio of 1:1, and numbers 5-10 are three sorghum varieties mixed at a ratio of 1:1:1.

[0097] Figure 14 Only 195bp bands were amplified in the two mixed samples of No. 1 and 3, the 190bp band in No. 2 was the result of Jinza 22 marker, the 190bp band in No. 4 was the amplification result of Jinza No. 103, and No. 5 was red. Both Yingzi and Jinza 22 were able to simultaneously amplify 190bp and 195bp bands. The 190bp and 195bp bands in No. 6 are the marker results of ...

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Abstract

The invention belongs to the field of molecular biology, and relates to a method for identifying the purity of varieties and germplasm of conventional varieties of wine making glutinous sorghum. The method includes synthesizing specific marker primers on the basis of basic principles of genome simple sequence repeats (SSR) according to information provided by databases; amplifying and screening molecular marker sites by the aid of extracted DNA (deoxyribonucleic acid) of varieties of sorghum to ultimately obtain genome specific bands of varieties of the 'red tassel' wine making glutinous sorghum and the like. The method has the advantages that sampling analysis and statistics are carried out according to maps of the specific bands of the varieties of the sorghum, and accordingly the purity of germplasm of raw materials for wineries can be identified; the method is easy and convenient to implement and high in feasibility, and obvious effects can be realized by the method.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to the application research of a genomic molecular marker technology to identify raw materials of wine sorghum. The invention relates to a method that can be used for the identification of different wine sorghum varieties and germplasm purity. Background technique [0002] With the in-depth adjustment of the Chinese liquor brewing industry, sorghum for winemaking has developed rapidly. In recent years, the demand for Maoxiang wine sorghum raw materials is very large, of which the "Red Tassel" sorghum in Guizhou Province has the largest promotion area. How to judge the authenticity of special sorghum germplasm is an important problem that the winery needs to face in the actual raw material purchase. In the past, the identification of raw materials in wineries was often based on the content of physical and chemical indicators (such as total starch, amylopectin, tannin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 闫松显王莉涂佑能汪地强王和玉赵亮
Owner KWEICHOW MOUTAI COMPANY
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