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Molecular marker of corn Opaque1 gene and application thereof

A technology of molecular markers and genes, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of inability to assist selection and obtain co-dominant DNA molecular markers, and achieve high accuracy, Simple operation effect

Inactive Publication Date: 2014-04-02
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have not obtained O1 The co-dominant DNA molecular markers on the gene and completely linked to it, so it is impossible to perform DNA molecular marker-assisted selection on the target traits controlled by the gene.

Method used

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  • Molecular marker of corn Opaque1 gene and application thereof
  • Molecular marker of corn Opaque1 gene and application thereof
  • Molecular marker of corn Opaque1 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: use o1 Mutant (o1 / o1) breeding high oil and high lysine maize

[0029] Compared with the corresponding wild type, o1 The total fatty acid content of the mutant grain increased by about 10%, and the three major components C16:0, C18:1, and C18:2, which accounted for about 90% of the corn fatty acid composition, had significant differences (Table 1, figure 1 ). Bundle o1 The mutant homozygous (o1 / o1) was crossed with other maize with excellent production traits, and then the F1 generation was selfed, and the recessive homozygote for high oil and high lysine corn kernels was isolated in the F2 generation. The recessive homozygote undergoes multiple generations of selfing, recurrent selection, and selects seeds with good grain development and high lysine content generation by generation, and finally obtains a stable recessive homozygote, high oil and high lysine inbred line or population, That is, high-oil and high-lysine commercial corn can be obtained. ...

Embodiment 2

[0032] Example 2: Obtaining and co-dominant molecular markers O1 Gene Linkage Determination

[0033] figure 2 and image 3 Outlines of wild-type (+ / +) and homozygous mutant (o1 / o1) corn kernels. F1 (o1 / +) was obtained by crossing the homozygous mutant (o1 / o1) with the homozygous wild type (+ / +), and then backcrossed with F1 as the female parent and the homozygous mutant as the male parent to construct a backcross ( BC) groups, see Figure 4 . Genotype segregation occurred in the BC population, including o1 / + and o1 / o1 genotypes, see Figure 5 , the grains with o1 / + genotype and + / + genotype are non-Opaque wild-type grains, and the grains with o1 / o1 genotype are Opaque mutant types.

[0034] The present invention is known by classical genetic mapping O1 The gene was located on the long arm of maize chromosome 10 (Bin4.07). through the O1 The backcross population (BC) was constructed with homozygous mutant and homozygous wild-type plants near the site, and the genomic ...

Embodiment 3

[0039] Example 3: Polymorphism identification of molecular markers among different parents

[0040] Extract the genomic DNA of inbred lines widely used in 10 kinds of breeding such as o1 / o1 mutant, wild type pure and B73, Mo17, W22, W64A, BSSS53, etc., and analyze the polymorphism among different varieties by PCR reaction . The results show that using the molecular marker MYO-PD in the present invention to o1 PCR amplification and gel electrophoresis analysis with 10 breeding parental DNAs including B73, Mo17, W22, W64A, BSSS53 showed that MYO-PD was o1 It has polymorphisms with 10 breeding parents such as B73, Mo17, W22, W64A, BSSS53, etc., see Figure 7 . This result demonstrates that the molecular marker MYO-PD can be accurately detected in any breeding background population o1 The presence of genes that can be used when breeding high oil and high lysine corn with these breeding materials o1 Assisted selection of genes with a false selection rate of 0.

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Abstract

The invention relates to a molecular marker of a corn Opaque1 gene and application thereof. The molecular marker is a codominant molecular marker MYO-PD obtained by amplifying specific primers through PCR (polymerase chain reaction), wherein the specific primers are base sequences shown as SEQ ID NO.1-SEQ ID NO.2. The molecular marker completely linked with an o1 gene can be used to perform genotype identification on DNA (deoxyribonucleic acid) of a corn at any period and in any tissue, thus detecting the existence of the o1 gene in each individual plant of the hybridization breeding filial generation and the heterozygous or homozygotic state thereof. Meanwhile, the molecular marker can be used to distinguish between mutants and 10 main breeding parents in China and can accurately detect the existence of the o1 gene in any breeding background population, and the mischoosing rate is 0. The detection method is high in accuracy and simple to operate, thus providing an important technical means for o1 gene-based DNA molecular marker aided breeding.

Description

technical field [0001] The present invention relates to a kind of corn Opaque1 Molecular markers of genes and their applications. technical background [0002] The world is experiencing ongoing shocks: food crises, spiraling energy consumption, increased climate warming and, most recently, economic recession. Riots triggered by the food crisis have occurred on every continent. Global demand and consumption of crops, an important source of food, feed and fuel, are increasing dramatically. In particular, dietary adjustments in emerging countries: rising meat consumption; and accelerated use of food for biofuel production in developed countries. This makes the food problem increasingly prominent, and has become one of the key factors restricting the development of the world economy and security. Therefore, only by continuously increasing the yield of crops and producing more food on the world's limited arable land can we meet social needs, maintain world security and mainta...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 王桂凤王芳钟鸣宇王佳佳王刚宋任涛许政暟
Owner SHANGHAI UNIV
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