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InDel molecular marker related to barley beer turbid character and application thereof

A molecular marker and barley technology, applied in the field of molecular biology and genetic breeding, can solve the problems of huge seed quantity, cumbersome measurement and analysis, and difficult to be accurate and reliable results

Inactive Publication Date: 2020-09-22
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods all need to collect a large number of barley seeds, and then carry out long-term malting, beer making and other steps, and also need to add cumbersome determination and analysis.
Therefore, the traditional method for measuring and analyzing the turbidity characteristics of barley beer is cumbersome and complicated, requires a huge amount of seeds, and the results are difficult to be accurate and reliable.

Method used

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  • InDel molecular marker related to barley beer turbid character and application thereof
  • InDel molecular marker related to barley beer turbid character and application thereof
  • InDel molecular marker related to barley beer turbid character and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Test materials

[0034] 120 DH lines constructed using both cultivated barley Franklin and Yerong were used as materials. The 120 materials were cultivated uniformly to harvest the seeds, dried and stored for later use.

[0035] (2) Character determination

[0036] The harvested barley seeds are subjected to mini-malting according to the following method: Weigh 200 grams of seeds, clean them, and then soak for 5 hours, 8 hours without water, 8 hours, 12 hours, 3 hours, and 1 hour at 15°C. Sprout again 82 hours later. After the germination is over, the drying process is carried out. After the dried malt was rooted, the winemaking experiment was carried out.

[0037] The wine-making test methods are as follows: (1) Weigh 50g of malt and smash it into 10 mesh size; (2) Saccharify the malt meal in a saccharification apparatus according to the protocol saccharification method; (3) Dilute according to the feeding ratio of 1:8; 4) Adjust the pH of the wort to about 5.4 with ph...

Embodiment 2

[0043] (1) Test materials

[0044] Twenty DH lines constructed using both cultivated barley Franklin and Yerong were used as materials, including 10 high-turbidity germplasm materials and low-turbidity germplasm materials, as shown in Table 1.

[0045] Table 1 20 barley germplasm materials with different turbidity characteristics

[0046]

[0047]

[0048] The CDS sequence alignment of the hazy1 gene of the above 20 barley materials showed that the CDS start site of the hazy1 gene of the low turbidity barley materials contained CCGCTG 70 bp downstream of the ATG, while the sequence of the high turbidity barley materials did not contain CCGCTG. .

[0049] (2) Acquisition of hazy1-InDel mark

[0050] According to the above 6bp deletion site, a pair of Indel labeled primers are designed, the upstream primer is 5'-CCGTCATGGTCTCCGTCTTC-3', and the downstream primer is 5'-AGTGTCCGAGCAGATTGGTC-3'.

[0051] (3) DNA extraction

[0052] Using fresh leaves at the seedling stage as materials, the CT...

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Abstract

The invention discloses an InDel molecular marker related to barley beer turbid characters and application thereof. The InDel molecular marker is designed based on a segment of 6bp deletion existing at 70bp downstream of an initial site ATG of a barley beer turbid gene hazy1 coding region. The method is used for identifying the beer turbidity character of barley. A nucleotide sequence of a codingregion of the barley beer turbid gene hazy1 is shown as SEQ ID NO. 1; the 6 bp deleted nucleotide sequence is CCGCTG. The invention discloses the InDel molecular marker closely linked with the barleybeer turbid character QTL for the first time, and the molecular marker is a codominant marker, is accurate and efficient in detection and convenient and stable in amplification, can be used for molecular marker-assisted selection, and improves the identification efficiency of barley varieties with different turbid characteristics.

Description

Technical field [0001] The present invention relates to the technical field of molecular biology and genetic breeding, in particular to an InDel molecular marker related to barley beer turbidity and its application. Background technique [0002] Barley (Hordeum vulgare L.) is the fourth largest cereal crop in the world after corn, wheat and rice in terms of planting area and output. It has a wide range of uses, mainly for food, feed and beer brewing. As the main raw material for brewing beer, barley's quality directly affects the quality and value of beer. Beer is a colloidal solution with complex ingredients and weak stability, which is prone to turbid precipitation during storage and transportation. Beer turbidity is a serious quality problem because it significantly shortens the storage time and shelf time of beer. With the increasing expansion of the beer industry and consumption, the turbidity problem has become increasingly prominent. Therefore, solving the problem of be...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 叶玲珍沈秋芳戴飞张国平
Owner ZHEJIANG UNIV
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