Molecular marker related to dwarfing of corn plant

A molecular marker, corn technology, applied in the field of molecular genetics, to achieve the effect of shortening the breeding cycle

Active Publication Date: 2021-10-08
QILU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on dwarf and semi-dwarf materials based on the B73 inbred line with the most thorough genome analysis.

Method used

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  • Molecular marker related to dwarfing of corn plant
  • Molecular marker related to dwarfing of corn plant
  • Molecular marker related to dwarfing of corn plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Molecular identification of the e975 mutant

[0028] According to the EcMutMap gene cloning method (principle see figure 1 A), select 35 individual plants with e975 mutant phenotype from the F2 segregation population prepared by wild-type B73 and e975 mutant, and extract DNA from the leaves. Karroten kit (Karroten, sK2304) is used for DNA extraction. For specific steps, refer to manual. Then 500ng of each individual plant was mixed in equal amounts and sent to the company for exon capture sequencing. Taking the B73 genome published online as a reference, GATK (v2.1-9) was used for variation mining. SnpEff v3.6c was used to annotate variant sites. The index (SNP-InDex) of single nucleotide substitutions (SNPs) in the mutation results was calculated using a self-written Perl script. The variation linked to the mutant phenotype is selected according to the following conditions: 1) G→A(C→T); 2) SNP-InDex>=0.9; 3) The variation type is non-synonymous mutation...

Embodiment 2

[0029] Example 2: Molecular marker design of e975 mutation site

[0030] The genome sequence of 250 bp was intercepted on both sides of the mutation site and used as a template for primer synthesis (as shown in SEQ ID NO.1-2, including a total of 501 bp of the mutation site), and Primer Premier 5 was used to design around the 251st base of the template Forward primer and reverse primer. Primer design principles: Tm value is around 60°C, product size is 250-550bp, and primer length is 22-24bp.

Embodiment 3

[0031] Example 3: Verification of wild-type and e975 mutant molecular markers

[0032] The primers were synthesized by Nanjing Qingke Biotechnology Co., Ltd., and the specific sequences are as follows.

[0033] Forward primer: 5'-CGATTTGCTGTAGAGGAACTCAGG-3'(SEQ ID NO.3),

[0034] Reverse primer: 5'-AATCAAACTTTCCGCCGAAGGTGT-3' (SEQ ID NO.4).

[0035] The total volume of the PCR system is 25 μL, including 1 μL of forward and reverse primers (10 μmol / L), 1 μL of genomic DNA, 0.2 μL of EasyTaq DNA polymerase, 2.5 μL of buffer, 2 μL of dNTPs, ddH 2 O to make up to 25 μL.

[0036] The PCR reaction program was: pre-deformation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 56°C for 30 s, and extension at 72°C for 30 s, a total of 35 cycles; over-extension at 72°C for 5 min; storage at 16°C.

[0037] The length of the PCR amplification product is 298bp, and then it is separated by 1% agarose gel electrophoresis, and the PCR product that meets the target fragment siz...

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Abstract

The invention discloses a molecular marker related to dwarfing of corn plant. According to the invention, a mutant e975 related to dwarfing of corn plant height is screened, although the section number of corn is obviously reduced, the height of the corn is obviously reduced, and the ear length of the corn is obviously reduced, the yield of the e975 is not obviously different from that of a wild type. Gene localization shows that the first exon of the Zm00001d027807 gene of the mutant has changed from G to A, so that non-polar glycine is changed into alkaline arginine. For the variation, a corresponding molecular marker primer (SEQ ID NO.3-4) is developed, and after PCR amplification, Sanger sequencing is utilized for genotype interpretation, so that the wild genotype and the mutant genotype of the filial generation contain corn dwarfing gene Zm00001d027807 can be identified. The molecular marker is beneficial to auxiliary breeding of new corn dwarfing varieties.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a molecular marker related to the dwarfing of maize plants. Background technique [0002] Corn has long been a basic food source for human existence and a major animal feed ingredient. my country's corn production ranks second in the world, and the increase in corn production is of great significance for solving my country's food, food, energy and other issues. One way to increase crop yield is to increase the yield per plant, and the other is to increase planting density. In recent decades, planting density has played a major role in improving maize yield (Duvick, 2005, Advances in agronomy 86, 83-145), and increasing planting density has become an important research direction for improving yield (Kebrom & Brutnell, 2007). The increase of maize planting density will lead to problems such as lodging, disease and photosynthetic efficiency reduction; dwarf varieties can improve ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6869C12N15/11A01H1/04
CPCC12Q1/6895C12Q1/6869A01H1/04C12Q2600/156C12Q2600/13C12Q2531/113C12Q2535/101Y02A40/146
Inventor 秦莉路小铎刘基生李海燕
Owner QILU NORMAL UNIV
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