Stem WSC Content Gene Marker Based on Zhongmai 895 and Its Application

A stem and content technology, applied in the field of WSC content gene markers and applications based on Zhongmai 895, can solve the problem of large genetic improvement space and other issues

Active Publication Date: 2021-10-08
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lot of dry matter in the stalks of current wheat varieties in the middle and late stages of grain filling, and there is still a lot of room for genetic improvement.

Method used

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  • Stem WSC Content Gene Marker Based on Zhongmai 895 and Its Application
  • Stem WSC Content Gene Marker Based on Zhongmai 895 and Its Application
  • Stem WSC Content Gene Marker Based on Zhongmai 895 and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Acquisition of the tightly linked molecular marker AX-111189530 and its KASP primer set at the QTL site QWsc.caas-1RS for WSC content in wheat stalks

[0072] 1. Acquisition of DH groups

[0073] Zhongmai 895 and Yangmai 16 are one of the main cultivars in the Huanghuai wheat region and the winter wheat region in the middle and lower reaches of the Yangtze River, respectively, with excellent comprehensive agronomic characteristics. Zhongmai 895 is a semi-winter wheat variety selected by the Institute of Crop Science of the Chinese Academy of Agricultural Sciences and the Cotton Research Institute of the Chinese Academy of Agricultural Sciences. In 2012, it passed the national examination and approval of Huanghuai South. Zhongmai 895 showed fast grain filling rate in the field, less WSC retention in the later stage, and high yield.

[0074] Taking Yangmai 16 as the female parent and Zhongmai 895 as the male parent, a DH population was constructed, including 1...

Embodiment 2

[0097] Embodiment 2, establishment of method

[0098] 1. Extract the genomic DNA of the leaves of the wheat to be tested, and dilute to obtain a template solution. The DNA concentration in the template solution was approximately 30 ng / μL.

[0099] 2. Carry out KASP.

[0100] Primer working solution: take primer A, primer B and primer C, make up to 100 μL with sterile ultrapure water, and mix well. In the primer working solution, the concentrations of primer A, primer B and primer C were 12 μM, 12 μM and 30 μM in sequence.

[0101] KASP 2×Master Mix is ​​a product of LGC (article number KBS-1016-002). KASP 2×Master Mix contains fluorescent probe A, fluorescent probe B, quenching probe A, quenching probe B, high-fidelity Taq enzyme, dNTP, Mg 2+Wait. The sequence of fluorescent probe A is 5'-GAAGGTGACCAAGTTCATGCT-3', and the 5' end is connected with a FAM fluorescent group. The sequence of fluorescent probe B is 5'-GAAGGTCGGAGTCAACGGATT-3', and the 5' end is connected with ...

Embodiment 3

[0114] Example 3. Using KASP primer set to detect genotypes based on specific SNPs in Yangmai 16, Zhongmai 895 and some DH population families

[0115] The wheats to be tested are: Yangmai 16, Zhongmai 895, and some DH population families obtained in Step 1 of Example 1. Yangmai 16 is a sample whose genotype is CC based on a specific SNP that has been verified by sequencing, and the genomic DNA of Yangmai 16 is used as a CC genotype control sample. Zhongmai 895 is a sample whose genotype is TT based on the specific SNP that has been verified by sequencing, and the genomic DNA of Zhongmai 895 is used as a control sample of TT genotype. Genomic DNA of Yangmai 16 and Genomic DNA of Zhongmai 895 were mixed in equal amounts to obtain a control sample of heterozygous genotype. Sterile ultrapure water was used as a blank control sample.

[0116] The method established in Example 2 was used to detect the genotype of the wheat to be tested based on the specific SNP.

[0117] see res...

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Abstract

The invention discloses a stem WSC content gene marker based on Zhongmai 895 and its application. The present invention provides a specific primer set, which consists of primer A shown in sequence 1, primer B shown in sequence 2 and primer C shown in sequence 3. The invention also protects a method for identifying the WSC content character of the stalk of the wheat to be tested. Detect the genotype of the tested wheat based on the specific SNP site. Using the genomic DNA of the wheat to be tested as a template, KASP was performed using a specific primer set, and fluorescence scanning was performed to determine the genotype of the wheat to be tested based on the specific SNP site. The stem WSC content of TT genotype was higher than that of CC genotype. The specific SNP sites are: the 36th nucleotide of the DNA molecule shown in Sequence 4 of the sequence listing in the wheat genome; the 223rd nucleotide of the DNA molecule shown in the Sequence 6 of the sequence listing in the wheat genome. The invention provides a good tool for breeding and utilizing the main QTL site QWsc.caas-1RS of WSC content in stems.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a gene marker for stem WSC content based on Zhongmai 895 and its application. Background technique [0002] The grain filling period is a critical period for determining wheat yield. my country's main wheat producing areas are facing severe dry and hot wind hazards in the middle and late stages of grain filling, and in severe cases, the yield reduction can reach more than 10%. The carbon source of wheat at grain filling stage mainly comes from photosynthetic products produced by flag leaves and water soluble carbohydrates (Water soluble carbohydrates, WSC) stored in stems and leaf sheaths. The contribution of culm WSC to yield is 10%-20% under normal water conditions, and up to 30%-50% under drought stress. During the evolution of wheat varieties, leaf photosynthetic efficiency and stalk WSC storage have been significantly improved, which has made important contributions to improvi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 肖永贵付路平徐小婷夏先春何中虎
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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