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Primer pairs used for identification of wheat vernalization gene VRN-1 and application thereof

The technology of VRN-A1 and VRN-B1 is applied to a primer pair set for identifying wheat vernalization gene VRN-1 and its application field, which can solve the problems of cumbersome detection procedures and high experimental cost, and achieves strong targeting and repeatability. Good sex, short time-consuming effect

Inactive Publication Date: 2013-02-06
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, to detect the allelic variation composition of the three loci of the vernalization gene VRN-1 in one wheat material, seven consecutive PCR tests are required, the detection procedures are cumbersome, and the experimental cost is high

Method used

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  • Primer pairs used for identification of wheat vernalization gene VRN-1 and application thereof
  • Primer pairs used for identification of wheat vernalization gene VRN-1 and application thereof
  • Primer pairs used for identification of wheat vernalization gene VRN-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, multiplex PCR detection of wheat vernalization gene VRN-A1

[0050] The multiplex PCR primer pair set of this example consists of two primer pairs specific to the VRN-A1 gene. The two primer pairs specific to the VRN-A1 gene are primer pair 1 composed of two single-stranded DNAs shown in sequence 1 and sequence 2 in the sequence listing, and two single-stranded DNAs shown in sequence 3 and sequence 4 Primer pair 2 consisting of DNA.

[0051] The multiplex PCR primer pair set in this example can simultaneously detect 4 genotypes of the VRN-A1 gene locus, namely three dominant allelic variant genes Vrn-A1a, Vrn-A1b, Vrn-A1c and one recessive allele Variant gene vrn-A1. For materials containing Vrn-A1a gene, two bands of 715bp and 624bp were simultaneously amplified by PCR; for materials containing Vrn-A1b gene, a band of 473bp was amplified by PCR; for materials containing Vrn-A1c gene, PCR was amplified A band of 493bp was obtained, but no band of 1068bp...

Embodiment 2

[0071] Example 2, multiplex PCR detection of wheat vernalization gene VRN-B1

[0072] The multiplex PCR primer pair set of this example consists of two primer pairs specific for the VRN-B1 gene. The two primer pairs specific to the VRN-B1 gene are primer pair 3 composed of two single-stranded DNAs shown in sequence 5 and sequence 6 in the sequence listing, and two single-stranded DNAs shown in sequence 5 and sequence 7 Composition of primer pairs 4.

[0073] The multiplex PCR primer pair set in this example can detect two genotypes of the VRN-B1 gene locus, namely, the dominant allelic variant gene Vrn-B1 and the recessive allelic variant gene vrn-B1. For the material containing the Vrn-B1 gene, a 709bp band was amplified by PCR; for the material containing the vrn-B1 gene, a 1149bp band was amplified by PCR.

[0074] 1. Multiplex PCR amplification of vernalization gene VRN-B1 in wheat varieties

[0075] 1. Preparation of wheat genome

[0076] Genomic DNA was extracted fro...

Embodiment 3

[0093] Example 3, multiplex PCR detection of wheat vernalization gene VRN-D1

[0094] The multiplex PCR primer pair set of this example consists of two primer pairs specific for the VRN-D1 gene. The two primer pairs specific to the VRN-D1 gene are primer pair 5 composed of two single-stranded DNAs shown in sequence 8 and sequence 9 in the sequence listing, and two single-stranded DNAs shown in sequence 8 and sequence 10 Composition of primer pairs 6.

[0095] The multiplex PCR primer pair set in this embodiment can detect two genotypes of the VRN-D1 gene locus, namely, the dominant allelic variant gene Vrn-D1 and the recessive allelic variant gene vrn-D1. For the material containing the Vrn-D1 gene, a 1671bp band was amplified by PCR; for the material containing the vrn-D1 gene, a 997bp band was amplified by PCR.

[0096] 1. Multiplex PCR amplification of vernalization gene VRN-D1 in wheat varieties

[0097] 1. Preparation of wheat genome

[0098]Genomic DNA was extracted ...

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Abstract

The invention discloses primer pairs for the identification or the assistant identification of the wheat vernalization gene VRN-1 and an application thereof. The primer pair groups provided by the invention comprise a primer pair group A, a primer pair group B and a primer pair group C, wherein four primers in the primer pair group A comprises DNA single chains represented by a sequence 1, a sequence 2, a sequence 3 and a sequence 4; three primers in the primer pair group B comprise DNA single chains represented by a sequence 5, a sequence 6 and a sequence 7; and three primers in the primer pair group C comprise DNA single chains represented by a sequence 8, a sequence9 and a sequence 10. The primer pair groups provided by the invention do not have mutual inhibiting effect and mispairing among all primers and have reliable detection result, good repeatability, low cost, strong specificity and short detection time. The primer pairs have great significance in detecting the winter-spring characteristic of wheat varieties, further increasing the efficiency of breeding and enhancing the selection of breeding high generation. In addition, the primer pairs the wheat popularization and variety introduction process to be more targeted.

Description

technical field [0001] The invention relates to a primer pair set for identifying or assisting in identifying wheat vernalization gene VRN-1 and application thereof. Background technique [0002] Vernalization is one of the basic stages of wheat growth and development. The vernalization response of wheat is closely related to its growth period and adaptability, and is one of the main bases to guide wheat regionalization and new variety promotion, and is controlled by vernalization genes. Vernalization genes determine 70-75% of the whole growth cycle of wheat. Studies by Iwaki and Van Beem have shown that there are differences in the vernalization gene composition of wheat varieties in different regions of the world, reflecting the different requirements of different ecological types for the vernalization response of varieties. [0003] The protein encoded by the vernalization gene VRN-1 is a MADS-box transcription factor, which is induced by low temperature to promote flow...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A01H1/04A01H1/02
Inventor 穆培源张晓科相吉山张影全桑伟徐红军韩新年聂迎彬崔凤娟邹波
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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