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Corn nicosulfuron anti-infection molecular marker and application thereof

A nicosulfuron-resistant and nicosulfuron-resistant technology is applied in the field of molecular markers for nicosulfuron-resistant resistance in maize, which can solve the problems of heavy workload and low efficiency, and achieve good variety improvement effects

Inactive Publication Date: 2016-11-23
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Screening maize nicosulfuron-resistant materials in traditional breeding requires large-scale field selection, with heavy workload and low efficiency, which is difficult to meet the current needs

Method used

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  • Corn nicosulfuron anti-infection molecular marker and application thereof
  • Corn nicosulfuron anti-infection molecular marker and application thereof
  • Corn nicosulfuron anti-infection molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Application of maize nicosulfuron resistance-related molecular markers

[0030] 1. DNA extraction from maize leaves

[0031] Using conventional techniques to extract DNA from maize leaves;

[0032] 2. Target product amplification

[0033] Forward primer sequence: 5'-CGTCGCCTTCCTCTTCTTGG-3' (shown in SEQ ID NO: 7); reverse primer sequence: 5'-ATGGACTCGAAGCGCGGGCGGTTGGCGATGATC-3' (shown in SEQ ID NO: 8).

[0034] PCR amplification: the PCR amplification system is 20μL

[0035]

[0036] Amplification conditions:

[0037]

[0038]

[0039] A 294bp fragment can be obtained through the above amplification, and its nucleotide sequence is shown in SEQ ID NO:6.

[0040] Specific digestion of PCR products:

[0041] Enzyme digestion system 10μL:

[0042] Specific enzyme BclI: 0.3 μL

[0043] PCR product: 3 μL

[0044] 10×NE buffer: 0.7μL

[0045] DdH 2 O: 6μL

[0046] Enzyme digestion reaction conditions: add 0.3 μL of BclI-specific enzyme (commercial...

Embodiment 2

[0048] Example 2 Application of maize nicosulfuron-sensitive related molecular markers

[0049] 1. DNA extraction from maize leaves,

[0050] DNA was extracted from maize leaves by conventional techniques; the leaves were derived from maize nicosulfuron-resistant inbred lines and maize nicosulfuron-sensitive inbred lines respectively;

[0051] 2. Target product amplification

[0052] Forward primer sequence: 5'-GCGTGCTTCACGGAGCAGGAC-3' (shown in SEQ ID NO:3); reverse primer sequence: 5'-TGAGCACGCCGAGGGAGA-3' (shown in SEQ ID NO:4).

[0053] PCR amplification: the PCR amplification system is 20μL

[0054]

[0055] Amplification conditions:

[0056]

[0057]A 295bp fragment can be obtained through the above amplification, and its nucleotide sequence is shown in SEQ ID NO:2.

[0058] Specific digestion of PCR products:

[0059] Enzyme digestion system 10μL:

[0060] Specific enzyme AvaII: 0.3μL

[0061] PCR product: 3 μL

[0062] 10×NE buffer: 0.7μL

[0063] DdH 2...

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PUM

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Abstract

The invention relates to the technical field of plant gene engineering, and provides a corn nicosulfuron anti-infection molecular marker. The marker is positioned on an nsfy gene of a corn No.5 chromosome; through the application of the molecular marker, whether a corn nicosulfuron sensitive and resistant gene is contained in a maize variety or selfing system or not can be detected for accelerating the breeding process of the corn nicosulfuron resistant variety. The special corn nicosulfuron sensitive and resistant molecular marker is used, so that the screening is fast and precise; the environment influence is avoided; the selection target is clear; the production cost is reduced; the selection efficiency and the quality of the corn nicosulfuron resistant variety or variety system are greatly improved.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, and provides a maize nicosulfuron resistance molecular marker and an application thereof. Background technique [0002] Corn is one of the important food crops in my country. At present, my country's per capita arable land is only 0.093 hectares. With the reduction of land and fresh water resources for food production, the problem of food supply security has become increasingly prominent. In the production of corn, the herbicide nicosulfuron has the advantages of being the most widely used, the highest safety, the least dosage, and low price, and is widely loved by farmers and breeders. In recent years, it has been found that more and more maize inbred lines and hybrids are sensitive to nicosulfuron, which seriously affects the normal maize breeding and production. [0003] The sensitivity of maize nicosulfuron is controlled by recessive single gene nsfy, and its resistance is...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘保申张永中戚秀山徐长利洪翠萍赵燕董鲁朋
Owner SHANDONG AGRICULTURAL UNIVERSITY