Molecular marker SSR52 of wheat few-tillering gene Ltn3 and application thereof

A technology of molecular markers and wheat, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems of high selection difficulty, high cost, and long time, so as to improve the efficiency of selection and identification, The effect of high success rate and high accuracy

Inactive Publication Date: 2015-08-05
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Wheat yield traits are complex quantitative traits, which are controlled by multiple quantitative trait loci (Quantitative trait locus, QTL). The traditional breeding method has the problems of long time, large consumption, high cost and small results

Method used

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  • Molecular marker SSR52 of wheat few-tillering gene Ltn3 and application thereof
  • Molecular marker SSR52 of wheat few-tillering gene Ltn3 and application thereof
  • Molecular marker SSR52 of wheat few-tillering gene Ltn3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] This example is used to illustrate the methods for obtaining wheat oligo-tiller gene Ltn3 and molecular marker SSR52: (1) Using oligo-tiller wheat H461 as the female parent and wheat Chuannong 16 as the male parent to cross hybrids to obtain hybrid F1, F1 generation single plant F2 was obtained by crossing, and the F8 generation RIL population containing 223 lines was obtained by single-seed transmission method, and 188 lines were randomly selected to form the genetic mapping population.

[0049] (2) Use the CTAB method to extract the DNA of each line of the genetic mapping population, and select 90K covering the genomes of hexaploid wheat A, B, and D published on wheatgenomics (http: / / wheatgenomics.plantpath.ksu.edu / ) SNP chip, using the DNA of parents H461 and Chuannong 16 as templates, genotyping, and obtaining the genotype data of the RIL population. The band pattern of the parent H461 is marked as A, and the band pattern of the parent Chuannong 16 is marked as B. ...

Embodiment 2

[0063] 1.1 Extraction of DNA

[0064] H461, Chuannong 16, and Chuanmai 107 were selected as test materials, among which Chuannong 16 and Chuanmai 107 were multi-tiller varieties, and H461 was an oligo-tiller variety. Leaf DNA of wheat samples at three-leaf stage was extracted by CTAB method.

[0065] 1.2 Screening of primers for detection of wheat oligo-tiller gene Ltn3

[0066] 1.2.1 Primer design

[0067] (1) Obtain the SNP probe sequence in this segment, obtain more sequence information of the target segment through the genome sequencing data of IWGS Wheat China Spring, electronically extend the sequences corresponding to both ends of the SNP probe sequence, and perform sequence Analysis, looking for simple sequence repeats (Simple Sequence Repeats, SSR) by SSRHunter software.

[0068] (2) Design 61 pairs of common PCR primers (Table 1) for subsequent screening. PCR primers were designed using Oligo7.0 and primer5.0 software. PCR primer design criteria: primer length 1...

Embodiment 3

[0082] Example 3 Detects the applicability of SSR52-F / R in identifying the tillering ability of genetic populations

[0083] (1) F1 was obtained by crossing H461 as the female parent and Chuanmai 107 as the male parent, F1 was self-crossed to obtain F2, and the RIL verification population of the F7 generation was added to the F7 generation by single seed propagation.

[0084] (2) Extract the leaf DNA of the population lines at the three-leaf stage, including H461 and Chuanmai 107.

[0085] (3) With the DNA obtained in step (1) as a template, carry out PCR amplification with the PCR primers of the present invention (upstream primer sequence as shown in SEQ ID No.2, downstream primer sequence as shown in SEQ ID No.3) .

[0086] (4) PCR amplification reaction system: 5μl 10×PCR buffer, 1.5U Ex Taq TM DNA polymerase, 2mmol / L MgCl 2 , 0.2mmol / L dNTP, 150ng each of upstream and downstream primers, 100ng template DNA, and double distilled water to a total of 50μl.

[0087] (5) P...

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Abstract

The invention discloses a molecular marker SSR52 which is closely linked with wheat few-tillering gene Ltn3, wherein the nucleotide sequence of the molecular marker SSR52 is represented as the SEQ ID No.1. Genetic distance between the molecular marker and the wheat few-tillering gene Ltn3 is 0.3 cM. A test result proves that the molecular marker can accurately trace the wheat few-tillering gene and predict tillering characters of wheat, thereby further carrying molecular-designing breeding conveniently. The invention also discloses a method of identifying the molecular marker of the wheat few-tillering gene Ltn3. By means of the method, accuracy of tillering prediction can be increased, success rate of specific plant type breeding can be increased, and achievement of an object of increasing per unit area yield of wheat is accelerated.

Description

technical field [0001] The invention relates to the field of wheat molecular breeding, in particular to a molecular marker SSR52 of wheat oligo-tiller gene Ltn3 and an application thereof. Background technique [0002] Wheat is the second largest grain crop after rice in my country. The planting area over the years accounts for 22% to 30% of the cultivated land area and 22% to 27% of the total grain crop area. It is mainly distributed in Henan, Hebei, Shandong, Shanxi, Shaanxi, Jiangsu, Sichuan, Anhui and other provinces; the total output is more than 100 million tons, accounting for about 22% of the grain crop output, and it is the staple food of about half of the population in my country. Therefore, the breeding of high-yielding wheat varieties has always been the focus of breeders. [0003] Wheat yield traits are complex quantitative traits, which are controlled by multiple quantitative trait loci (Quantitative trait locus, QTL). The traditional breeding method has the pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 刘亚西王智强石浩然莫洪君卢艳丽甯顺腙江千涛魏育明郑有良
Owner SICHUAN AGRI UNIV
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