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Grain weight molecular marker for wheat and application of grain weight molecular marker in breeding

A heavy molecule, wheat grain technology, applied in the fields of genetic engineering and wheat breeding, can solve the problems of QTLs loci that have not been verified by the breeding process or variety validity, poor practicability, and poor accuracy.

Active Publication Date: 2014-10-08
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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  • Grain weight molecular marker for wheat and application of grain weight molecular marker in breeding
  • Grain weight molecular marker for wheat and application of grain weight molecular marker in breeding
  • Grain weight molecular marker for wheat and application of grain weight molecular marker in breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The leaf DNA of embodiment 1 wheat is extracted

[0023] (1) Take about 0.3-0.5 g leaves into a 5mL centrifuge tube, freeze them in liquid nitrogen and grind them into powder;

[0024] (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;

[0025] (3) Cool down to room temperature, wait for 10 minutes, add 10-15μL RNase (10mg / mL) in 37℃ water bath for 30 minutes, shake gently several times to fully mix, about once / 10 min;

[0026] (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 min, and place in a refrigerator at 4°C Let stand for 5 minutes, then centrifuge at 8000 rpm for 10 minutes;

[0027] (5) Take the supernatant in another tube, about 1300 μL, add an equal volume of cold chloroform (placed...

Embodiment 2

[0043] Embodiment 2 target product amplification

[0044] Forward primer sequence: 5'-CCTTCCATATGTTTTTTTAATGAGCCGCC-3' (as shown in SEQ ID NO:3)

[0045] Reverse primer sequence: 5'-GCTTCTTTTCCAACTCAATAAATGAGCC-3' (as shown in SEQ ID NO:4)

[0046] PCR amplification: the PCR amplification system is 20μL

[0047]

[0048]Note: Mix available: or (Taq enzyme 0.25μL, DNK 2.0μL, Buffer1.5μL, MgCl 0.4μL configuration.)

[0049] Amplification conditions:

[0050]

[0051] A 668bp fragment can be obtained through the above amplification, and its nucleotide sequence is shown in SEQ ID NO:2.

Embodiment 4

[0052] Example 4 Specific enzyme digestion of PCR products:

[0053] Enzyme digestion system 10μL:

[0054] Specific enzyme HaeIII: 0.3 μL

[0055] PCR product: 3 μL

[0056] 10×NE buffer: 0.7μL

[0057] wxya 2 O: 6μL

[0058] Enzyme digestion reaction conditions: add 0.3 μL of HaeIII specific enzyme (obtained from the market) to the PCR amplification product, bathe in water at 37°C for 2 hours, then inactivate the enzyme digestion system at 65°C for 5 min.

[0059] After the above-mentioned amplified products were separated by electrophoresis on 8% polyacrylamide gel, the molecular weight of the amplified product was 668bp. A 303bp electrophoresis band appeared. However, in varieties or lines with an increased thousand-grain weight gene, the segment is chopped and the segment is deleted.

[0060]

[0061]

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Abstract

The invention relates to the technical field of genetic engineering and can be applied to the field of wheat breeding, and particularly provides a grain weight molecular marker for wheat. The marker is located between RFL-CONTIG4632-1512 and TaGW2-CAPS of 6A chromosome of wheat. By applying the molecular marker, whether a wheat variety or strain has QGW6A-232 for increasing thousand seed weight or not can be detected so as to accelerate the breeding progress of high-yield variety of wheat. As the special grain weight molecular marker is adopted, screening becomes quick and accurate without being affected by the environment, the selected target is clear, moreover, the production cost is saved, so that the selecting efficiency and quality of the high-yield variety or strain of wheat 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. have shown that under the premise that the number of spikes and grains per spike are relatively fixed, the yield per hectare can increase by 157 kg 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 easi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 田纪春李青芳陈建省张莹邓志英陈广凤孙彩铃
Owner SHANDONG AGRICULTURAL UNIVERSITY
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