Primer, kit, detection method and application thereof for detecting low grain protein content of wheat

A protein content and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of low single effect value, low accuracy and breeding efficiency, and increase breeder traits. Select workload and other issues to achieve the effect of reducing workload, improving selection accuracy and breeding efficiency, and reliable methods

Inactive Publication Date: 2019-07-05
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Blanco et al. (2002) discovered seven QTL loci controlling grain protein content, located on chromosomes 4B, 5A, 6A, 6B, 7A, and 7B, respectively; Prasad et al. (1999) located the QTL loci controlling grain protein content in On chromosome 2D; Huang et al. (2006) used doubled haploid populations to find QTLs controlling grain protein content on chromosomes 4B and 4D; Li et al. (2007) used two recombinant inbred lines containing 229 and 485 families 3 and 10 QTLs affecting grain protein content were detected in the population, respectively located in 2B, 3A, 4A, 4D, 5B, 7A, 7B and 1A, 1B, 2A, 2D, 3A, 4B, 5A, 5D, 6B , 7D chromosome, according to the current public literature, there are many genes or QTLs that control the grain protein content, but the single effect value is not high, and the conventional method can only check the grain protein content after the 4th generation of hybridization , thus increasing the workload of breeders to select this trait, and the accuracy and breeding efficiency are low

Method used

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  • Primer, kit, detection method and application thereof for detecting low grain protein content of wheat
  • Primer, kit, detection method and application thereof for detecting low grain protein content of wheat
  • Primer, kit, detection method and application thereof for detecting low grain protein content of wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: the mensuration of wheat grain GPC

[0024] The 80 wheat varieties in Table 1 were used as experimental materials, and the GPC of wheat grains was determined by the method shown in GB 50095-2010. The GPC data is the average value of 80 wheat varieties planted between three points in two years, and the planting soil is consistent.

[0025] Table 1: Determination results of wheat GPC and PCR amplification bands

[0026]

[0027]

[0028]

[0029]

[0030] Note: "+" in the column of PCR amplification products in the table indicates that the corresponding target band was amplified, and "-" indicated that the corresponding target band was not amplified.

Embodiment 2

[0031] Example 2: Synthesis and detection method of wheat grain GPC-related molecular markers

[0032] 1. Synthesis of wheat GPC-related molecular markers

[0033] A primer composed of sequence I and sequence II in the sequence table was synthesized, referred to as GPC-B.

[0034] The upstream and downstream primers in primer GPC-B are as follows:

[0035] GPC-B-F (sequence I): 5'-CTACCGTACTCTCTCTGATT-3' (position 1-20 of sequence III);

[0036] GPC-B-R (sequence II): 5'-TGGATCTCGCAGCAAGTTCT-3' (reverse complement sequence of positions 1992-2013 of sequence III).

[0037] 2, Utilize the method for the molecular marker detection wheat GPC content of step 1

[0038] (1) PCR amplification

[0039] Using wheat genomic DNA as a template, PCR was carried out with primer GPC-B respectively.

[0040] Reaction system (20μl): 2.0μL 10×PCR buffer, 50ng / μL DNA 1.0μL, 2.5mmol / L total dNTPs 2.0μL, upstream primer (GPC-B-F) with a concentration of 10μmol / L 0.20μL, a concentration of 10μ...

Embodiment 3

[0046] Embodiment 3: Primer GPC-B is used to detect the effectiveness verification of wheat low GPC

[0047] A total of 80 wheat varieties in Table 1 were used for PCR analysis with primer GPC-B, and the operation method was the same as step 2 in Example 2. According to the electrophoresis result of the PCR amplification product, the result is judged with reference to the result judgment standard in Step 2 2 (3) of Example 2.

[0048] The results are shown in Table 1, and the agarose gel detection results of some wheat varieties are as follows figure 2 As shown, it can be seen from the results that among the 80 wheat varieties, there were 38 wheat varieties that obtained the target band (sequence III) with a size of 2013bp when using primer GPC-B to carry out PCR amplification. The range and mean value are shown in Table 1. The average value between (10.2%-15.2%) is 12.6%; there are 42 wheat varieties that have not amplified the target band with an average size of 2013bp, an...

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PUM

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Abstract

The invention discloses a primer, a kit, a detection method and an application thereof for detecting low grain protein content of wheat; the upstream primer sequence of the primer is shown in SEQ ID NO: I, and the downstream primer sequence is shown in SEQ ID NO: II Show. The method for detecting low GPC in wheat provided by the present invention is reliable, convenient and practical, has important application prospects in molecular marker-assisted selection of wheat germplasm resource evaluation and breeding, and provides a reference for identifying wheat varieties with low grain protein content in accordance with. The invention can screen low grain protein content in the 2nd generation of breeding hybridization, overcomes the conventional method and can only screen low grain protein content after the 4th generation of hybridization, thus reducing the work of breeders to select the trait It is an efficient and fast breeding new technology to improve the accuracy of selection and breeding efficiency.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer, a kit, a detection method and an application thereof for detecting low grain protein content of wheat. Background technique [0002] With the continuous increase of my country's wheat production, people's demand for high-quality wheat is increasing. According to the quality of wheat, my country divides wheat into strong gluten wheat, medium gluten wheat and weak gluten wheat. Weak gluten wheat is mainly used for the production of biscuits, pastries and For flour products such as southern steamed buns, wheat grain protein content (GPC) is an important indicator of quality. Wheat varieties with low grain protein content can be used as raw materials for the production of high-quality weak-gluten wheat products. my country’s national standard for high-quality weak-gluten wheat (GB / T17893- 1999) stipulates that the crude protein content of wheat grain shall not exceed ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 冯波王涛金秀锋徐智斌樊小莉刘静
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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