SSR primer group and method for malt variety identification by virtue of primer group

Abstract

The invention provides an SSR primer group, which comprises 6 pairs of SSR primers, namely scssr07759, HVM68, Bmac0030, scssr10148, scssr03907 and Bmag0120. The invention also provides a method for malt variety identification by virtue of the SSR primers, wherein the method comprises the following steps: extraction of DNA of a sample of experimental variety, PCR amplification by virtue of the SSR primers, polyacrylamide gel electrophoresis as well as silver staining and SSR spectrum band analysis. The method, through SSR spectrum band analysis after the PCR amplification of the 6 pairs of SSR primers, can be used for identifying experimental malt variety and known malt variety and can be used for constructing a malt variety database. Through malt variety identification by virtue of the specific SSR primer group, the method can be used for rapidly identifying various malt varieties, and the method has the advantages of being rapid and accurate, and convenient in operation.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method for identifying malt varieties, in particular to a technique for identifying malt varieties using SSR primers. Background technique [0002] Beer is a low-alcohol alcohol full of carbon dioxide, which is brewed by yeast fermentation with malt, hops, and water as the main raw materials. As the main raw material of beer, malt is made by soaking, sprouting, drying and roasting barley. The quality of malt directly determines the quality of beer. Variety identification of barley and malt is the most important indicator for evaluating barley quality and malt quality, both of which are important indicators for ensuring beer quality. [0003] At present, the identification technology of barley varieties has been relatively mature, including traditional morphological identification methods, protein electrophoresis identification methods and DNA molecular marking methods. The co...

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Problems solved by technology

[0004] Compared with the relatively mature technology of barley variety identification, there is no clear detection method for malt variety identification. Conventional malt indicators are indicators that reflect the mixed state of malt and cannot reflect changes in malt varieties.

However, the existing methods for identification of barley varieties are not suitable for the identification of malt varieties.

(1) The appearance characteristics of barley have changed after germination and drying, and the protein begins to decompose under the action of protease, so the traditional barley morphology identification method and protein electrophoresis identification method are completely unsuitable for the identification of malt varieties

(2) In terms of molecular marker technology, barley DNA is usually extracted from germinated fresh seedling leaves. Fresh leaves contain less impurities and DNA extraction is easy; Made after the malting process, it cannot be germinated again, so DNA can only be extracted directly from the malt

Because malt is rich in proteins, polysaccharides, polyphenols and other substances, it is difficult to separate

(3) More critically, part of the DNA of the malt is broken after being roasted at 84-86°C for 3 hours, which directly affects the analysis of the results of PCR amplification fragments, resulting in many primers that play a decisive role in the identification of barley varieties being amplified by PCR. The shape and position of the electrophoresis bands on the polyacrylamide gel electrophoresis map of the increased products are basically the same, and the corresponding malt varieties cannot be distinguished

[0005] Due to the lack of technical means for identifying malt varieties, beer companies not only suffer huge economic losses when purchasing malt, but also the poor uniformity of adulterated malt directly affects the performance of saccharification, filtration and brewing, reduces the quality of beer, and ultimately affects the brand of beer companies image

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