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Wheat grain weight molecular marker and its use in breeding

A heavy molecule, wheat grain technology, applied in the field of genetic engineering and wheat breeding, can solve the problems of reducing the actual efficiency of molecular assisted selection, QTLs loci have not been verified by the breeding process or variety validity, and too many QTLs.

Active Publication Date: 2014-11-26
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to this standard, most of the published QTLs loci have not been validated by breeding process or varieties
[0006] ② Poor practicability of QTLs: The accuracy of the loci mapped by a single genetic population is poor, and due to the large confidence interval, too many QTLs or the existence of false positive QTLs, the actual efficiency of molecular assisted selection is reduced, and it cannot even be effectively applied to wheat breeding

Method used

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  • Wheat grain weight molecular marker and its use in breeding
  • Wheat grain weight molecular marker and its use in breeding
  • Wheat grain weight molecular marker and its use in breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The leaf DNA of embodiment 1 wheat is extracted

[0022] (1) Take about 0.3-0.5g leaves into a 5mL centrifuge tube, freeze them in liquid nitrogen and grind them into powder;

[0023] (2) Add about 1600 μL of buffer S that has been preheated to 65°C, mix by inversion several times, bathe in water for 0.5-1 hour, and shake gently several times during this period to fully mix;

[0024] (3) Cool down to room temperature, wait for 10 minutes, add 10-15μL RNase (10mg / mL) in a 37℃ water bath for 30 minutes, and shake gently several times to fully mix, about once / 10min;

[0025] (4) Take out the centrifuge tube, add an equal volume of 1600 μL, extract at 4°C with phenol (Tris-balanced phenol):chloroform:isoamyl alcohol (25:24:1 (volume ratio), mix gently for 10 minutes, and place in a refrigerator at 4°C. Place for 5min, then centrifuge at 8000rpm for 10min;

[0026] (5) Take the supernatant in another tube, about 1300 μL, add an equal volume of cold chloroform (placed in a ...

Embodiment 2

[0042] Embodiment 2 target product amplification

[0043] Forward primer sequence: 5'-CTACTTATGATTACTCATCCTCTGACT-3' (as shown in SEQ ID NO:3)

[0044] Reverse primer sequence: 5'-ACATCCCATTGACACAATAATCTGCTCTC-3' (as shown in SEQ ID NO:4)

[0045] PCR amplification: the PCR amplification system is 20μL

[0046]

[0047]Remarks: Mix available: or (Taq enzyme 0.25μL, DNK2.0μL, Buffer1.5μL, MgCl0.4μL configuration.)

[0048] Amplification conditions:

[0049]

[0050] A 952bp fragment can be obtained through the above amplification, and its nucleotide sequence is shown in SEQ ID NO:2.

Embodiment 4

[0051] Example 4 Specific enzyme digestion of PCR products:

[0052] Enzyme digestion system 10μL:

[0053] Specific enzyme ScrFI: 0.3 μL

[0054] PCR product: 3 μL

[0055] 10×NE buffer: 0.7μL

[0056] wxya 2 O: 6μL

[0057] Enzyme digestion reaction conditions: Add 0.3 μL of ScrFI specific enzyme (commercially available) to the PCR amplification product, bathe in 37°C water for 2 hours, then extinguish the enzyme digestion system at 65°C for 5 minutes.

[0058] After the above-mentioned amplified products were separated by electrophoresis on 8% polyacrylamide gel, the molecular weight of the amplified product was 952bp. A 125bp electrophoresis band appeared. However, in varieties or lines with an increased thousand-grain weight gene, the segment is chopped and the segment is deleted.

[0059]

[0060]

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Abstract

The invention relates to the technical field of gene engineering and provides a wheat grain weight molecular marker. The wheat grain weight molecular marker can be used in the field of wheat breeding. The wheat grain weight molecular marker is located between wheat 6A chromosome BOBWHITE_C20706_135 and RA_C28538_971. Through use of the wheat grain weight molecular marker, it is detected if the wheat variety or line comprises QGW6A-164 for increasing thousand seed weight so that wheat high-yield variety breeding is accelerated. Through use of the special-purpose grain weight molecular marker, screening is fast and accurate, is not influenced by the environment and has a clear selection object, a production cost is greatly reduced and high-yield wheat variety or line selection efficiency and quality are greatly improved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, can be applied to the field of wheat breeding, and specifically provides a wheat grain weight molecular marker. Background technique [0002] Wheat is the main ration of 50-60% of the population in my country. At present, the per capita arable land in my country is only 0.093 hectares. With the reduction of land and fresh water resources for food production, the problem of food supply security has become increasingly prominent. Increasing the thousand-grain weight is one of the important ways to cultivate high-yielding wheat First, Tian Jichun et al. showed that under the premise that the number of spikes and grains per spike are relatively fixed, the yield per hectare can increase by 157kg for every 1 gram increase in thousand-grain weight. [0003] Grain weight is jointly controlled by major genes and minor genes. The number of genes is large and the effect is low, which is easily af...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 陈建省田纪春李青芳刘凯孙彩玲王新霞
Owner SHANDONG AGRICULTURAL UNIVERSITY
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