Gene relevant to wheat thousand seed weight, functional marker and application thereof

A thousand-grain weight and genetic technology, applied in the field of wheat molecular biotechnology and breeding applications, can solve problems affecting wheat thousand-grain weight

Active Publication Date: 2014-05-28
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the influence of members of this gene family on thousand-grain weight of wheat

Method used

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  • Gene relevant to wheat thousand seed weight, functional marker and application thereof
  • Gene relevant to wheat thousand seed weight, functional marker and application thereof
  • Gene relevant to wheat thousand seed weight, functional marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Cloning of 1000-grain weight-related gene TaSnRK2.10 gene in wheat, the material used is Lumai 21.

[0050] 1) Electronic cloning of wheat TaSnRK2.10 cDNA sequence;

[0051] (1) Using the rice SAPK10 (Accession: AB125311) gene sequence submitted by NCBI as a probe, search the wheat EST database, splice the obtained sequences with DNAMAN software, and predict the open reading frame to obtain the complete wheat SnRK2.10cDNA sequence .

[0052] (2) According to the obtained full-length cDNA sequence, two pairs of specific primers SnRK2.10-1 and SnRK2.10-2 were designed and screened by Primer5 software, which were used to clone wheat SnRK2.10 cDNA and gDNA gene respectively.

[0053] TaSnRK2.10-1 forward primer: 5′-GCTTGCTCGGTTGCTTTGC-3′ (SEQ ID NO: 7)

[0054] Reverse primer: 5'-CATCCAAAAGGCCAAACCGT-3' (SEQ ID NO: 8)

[0055] TaSnRK2.10-2 forward primer: 5′-GTCAAGTACATCGAGCGAGGG-3′ (SEQ ID NO: 9)

[0056] Reverse primer: 5′-CGTCGTTCAGGAACTGGTTGA-3′ (SEQ ID NO: 10)

[...

Embodiment 2

[0082] Functional marker development and association analysis

[0083] 1) Use TaSnRK2.10-2 (forward primer sequence is SEQ ID NO: 9, reverse primer sequence is SEQ ID NO: 10) primer sequence to PCR amplify DNA from 10 materials in Huanghuai wheat region and analyze the genome sequence It was found that the TaSnRK-4A (SEQ ID NO: 4) sequence had SNP and InDel sequence differences among varieties.

[0084] 2) There are 3 SNP sites and one InDel site in the TaSnRK2.10-4A genome sequence, such as image 3 As shown in a: the 1123rd nucleotide from the 5' end is C or T, the 1321st nucleotide from the 5' end is A or T, the 1757th nucleotide from the 5' end is G or C, from Nucleotides 1832-1834 at the 5' end are deletions or insertions of TGA trinucleotides. Three SNP sites and one InDel site are closely linked: when the 1123rd nucleotide from the 5' end is C, the 1321st and 1757th nucleotides from the 5' end must be A and G, and at the same time The 1832-1834th nucleotide is the de...

Embodiment 3

[0100] Method for Detecting 1000-grain Weight of Wheat Varieties or Strains Using TaSnTK2.10-4A-caps Markers

[0101] 1) Extract the DNA of the wheat variety to be tested, and use TaSnTK2.10-4A-caps labeled primers (forward primer sequence: SEQ ID NO: 11, reverse primer sequence: SEQ ID NO: 12) to perform PCR on the wheat variety DNA Amplification, the PCR amplification system is 20 μl, including H 2 O 14.3 μl, 2×PCR buffer 2.0 μl, Taq enzyme 0.20 μl, left reverse primer 0.5 μl, cDNA 1.0 μl, dNTP 1.5 μl; amplification conditions are 94°C pre-denaturation for 4 minutes; 94°C denaturation for 40 seconds, 58°C Anneal for 45s, extend at 72°C for 1min, 35 cycles; extend at 72°C for 10min; store at 10°C.

[0102] 2) Digest the above-mentioned amplified products with SalI endonuclease respectively, and detect whether the SNP site at position 1757 of the 5' end of the wheat TaSnRK-4A genome sequence to be tested is C or G, and determine whether the genotype of the wheat to be tested ...

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Abstract

The invention discloses a gene TaSnRK2.10 relevant to the wheat thousand seed weight, a molecular marker TaSnTK2.10-4A-caps relevant to the gene and application of the marker. The DNA of a wheat variety to be detected is subjected to PCR (polymerase chain reaction) multiplication through a marker primer TaSnTK2.10-4A-caps; a multiplication product is subjected to cleavage through SalI incision enzyme and is also subjected to electrophoretic separation; for example, PCR products are two bands with the sizes of 793bp and 316bp; the wheat variety is a variety with haplotype with high thousand grain weight of the gene; the PCR product is only a band with the size of 106bp; the wheat variety is a variety which is not provided with the haplotype with high thousand grain weight of the gene. According to the gene TaSnRK2.10 relevant to the wheat thousand seed weight and the molecular marker TaSnTK2.10-4A-caps thereof, the wheat variety or strain with high thousand grain weight can be conveniently detected or screened, and the selection process of the high-yield variety of wheat can be greatly accelerated.

Description

technical field [0001] The invention relates to the fields of wheat molecular biotechnology and breeding applications, in particular to a gene related to wheat thousand-grain weight, molecular markers and applications thereof Background technique [0002] Wheat is one of the most important food crops in the world, so breeding high-yielding wheat varieties has always been highly valued by breeders. Studies have shown that the thousand-grain weight, which is mainly affected by the grain size trait, is the most stable direct component of yield. For every 1g increase in the thousand-grain weight of wheat, the yield of wheat per hectare will increase by 140-160kg (Tian et al. 2006). It can be seen that increasing the thousand-grain weight of wheat to increase yield is one of the most important breeding goals of wheat. [0003] Functional markers are a new type of molecular markers developed based on polymorphic motifs within functional genes that cause phenotypic traits to vary....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/11C12Q1/68
Inventor 李斯深张照贵赵岩孔凡美郭营
Owner SHANDONG AGRICULTURAL UNIVERSITY
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