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Molecular marker for identifying wheat leaf villous gene, primer set and application

A technology of molecular markers and primer sets, which is applied in the field of molecular biology, can solve the problems of unreported molecular markers for identification of wheat fluff genes, lack of in-depth research, and few studies on molecular genetic mechanisms, so as to eliminate the influence of environment and external factors, Improved utilization efficiency and clear location

Active Publication Date: 2021-08-20
CROP INST SICHUAN PROVINCE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, there are few studies on the molecular genetic mechanism of the leaf hairy gene, especially the molecular basis research such as the positioning of the relevant genes is not yet in-depth; there is no report about the molecular markers for identifying the wheat hairy gene, and there is no use of molecular markers to assist selection. Breeding selection of wheat fluff has not been reported

Method used

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  • Molecular marker for identifying wheat leaf villous gene, primer set and application
  • Molecular marker for identifying wheat leaf villous gene, primer set and application
  • Molecular marker for identifying wheat leaf villous gene, primer set and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] A molecular marker for the identification of the wheat leaf hair gene, which is located on the 7BS chromosome of W. pilosa, named QLP.saas-7BS (eg Figure 6 shown); the QLP.saas-7BS includes two linked molecular markers, the KASP marker of KASP-AX-86175290 and the KASP marker of AX-86172908, and the QLP.saas-7BS localization interval is AX-86175290 ( The physical location is 61106266-61106336b) and AX-86172908 (the physical location is 60629908-60629979b) in the physical interval of 0.48Mb.

Embodiment 2

[0060] The preparation method of molecular marker described in embodiment 1, technical route is as follows figure 1 Shown, its preparation comprises the following steps:

[0061] (1) Constructed the genetic population of high-generation recombinant inbred lines

[0062] The genetic population F of high-generation recombinant inbred lines was constructed by using the commonly cultivated hairless wheat Chuanmai 104 and the wheat landrace Baimai with high-density leaf hairs 8 generation;

[0063] Such as figure 2 As shown, the profile photos of Chuanmai 104 and white hairy wheat leaf fluff, Chuanmai 104 has no fluff, and white hairy wheat has long hair;

[0064] Such as image 3 As shown, the frontal photos of Chuanmai 104 and white hairy wheat leaves have no downy leaves, and white hairy wheat leaves have long and dense hairs;

[0065] (2) Construction of genetic linkage map

[0066] Using 50K high-density SNP chips to scan the entire genome of the population, construct a...

Embodiment 3

[0082] Verify the KASP marker of AX-86175290 and the KASP marker of AX-86172908 in Example 1, the KASP marker primer detection reagent uses the mix reagent of Aiji Analysis Technology (Shanghai) Co., Ltd., and the quantitative PCR instrument uses the CFX384 Real of BIO-RAD Company -Time System; the phenotypic identification of leaf villi is observed through a "stereo microscope LEICA E24HD"; the detection of specific markers is performed through a "fluorescence quantitative PCR instrument for typing detection; specifically include the following steps:

[0083] (1) Extract the wheat genomic DNA to be identified and distribute it on the PCR reaction plate;

[0084] (2) Use the labeled primer set KASP-AX-86175290 and KASP-AX-86172908 to perform PCR amplification on the DNA extracted in step (1);

[0085] The method of PCR amplification is: first configure the KASP genotyping mixture, add the configured KASP genotyping mixture to the PCR reaction plate filled with DNA in step (1),...

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Abstract

The invention provides a molecular marker for identifying a wheat leaf villous gene, a primer set and an application. The gene is a new gene, is positioned on a 7BS chromosome of a wheat local variety white wheat, and is named as QLP.saas-7BS. The molecular marker is positioned between an AX-86175290 probe and an AX-86172908 probe of an SNP (Single Nucleotide Polymorphism) chip, KASP markers are respectively developed according to probe sequences, and the KASP markers are respectively named as KASP-AX-86175290 and KASP-AX-86172908. According to the invention, the newly developed KASP molecular marker can effectively detect and screen the wheat villous character, and the molecular marker technology is applied to breeding the wheat variety with the leaf villous phenotypic character, so that the production cost is saved, the selection efficiency is improved, and the breeding cycle of the wheat variety is shortened; and the detection is convenient and rapid and is not influenced by the environment.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a molecular marker, primer set and application for identifying new genes of wheat leaf villi. Background technique [0002] The main function of wheat leaves is to carry out photosynthesis and produce organic matter, and it is also an important organ of wheat respiration and transpiration. There are 5 kinds of leaves of wheat, including scutellum (degenerate leaf), coleoptile (incomplete leaf), tiller sheath (incomplete leaf), shell (metamorphosis leaf) and green leaf (complete leaf). The leaves we usually refer to are fully developed green leaves. The leaf consists of the body (blade), pillow, sheath, auricle, and ligule. The connection between the blade and the sheath is called the leaf pillow. Leaf sheaths grow on the stem nodes and surround all or part of the internodes. The main function is to strengthen the stem strength and also perform photosynthesis. The ligule is lo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11A01H1/04
CPCC12Q1/6895C12Q1/6858A01H1/04C12Q2600/13C12Q2600/156C12Q2531/113C12Q2563/107Y02A50/30
Inventor 刘泽厚王琴万洪深杨凡李俊杨武云杨宁
Owner CROP INST SICHUAN PROVINCE ACAD OF AGRI SCI
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