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Molecular marker closely linked with corn bacterial wilt resistance genes and primer and application thereof

A molecular marker and resistance gene technology, applied in the field of agricultural biology, can solve the problems of low selection efficiency and long breeding cycle, and achieve the effect of reducing labor land, shortening breeding cycle, and improving corn yield level.

Active Publication Date: 2014-10-08
HEFEI FENGLE SEED
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  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Conventional breeding methods based on phenotypic selection have disadvantages such as low selection efficiency and long breeding cycle. It is urgent to inject modern molecular technology, supplemented by high-efficiency genotype-directed selection, in order to quickly and efficiently breed excellent new varieties of maize.

Method used

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  • Molecular marker closely linked with corn bacterial wilt resistance genes and primer and application thereof
  • Molecular marker closely linked with corn bacterial wilt resistance genes and primer and application thereof
  • Molecular marker closely linked with corn bacterial wilt resistance genes and primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1, Obtaining the special primer pair for the identification of bacterial wilt resistance traits: the nucleotide sequence shown in SEQ ID NO: 3 in the sequence table and the nucleotide sequence shown in SEQ ID NO: 4 in the sequence table

[0029] 1. Parental Genome Amplification

[0030] The parents used to create the maize recombinant inbred line (RIL) population are 1145 and 0908 selected by our company. 0908 is the female parent, which is not resistant to bacterial wilt; 1145 is the male parent, which is highly resistant to bacterial wilt.

[0031]Genomic DNA of parental leaves was extracted by CTAB method, using primers published by Qing Yang (Qing Yang, Zhi Li, et al. CACTA-like transposable element in ZmCCT attenuated photoperiod sensitivity and accelerated the postdomestication spread of maize. PNAS 2013, 10.) Yes, it is composed of the nucleotide sequence shown in SEQ ID NO: 5 in the sequence table and the nucleotide sequence shown in SEQ ID NO: 6 in the ...

Embodiment 2

[0038] Example 2, Correlation verification between the amplification product of QK1 and the resistance traits of corn bacterial wilt

[0039] 96 F 2 The substitute material is the test object. Genomic DNA of corn endosperm was extracted by alkaline boiling method, and the specific steps were the same as in Example 1. PCR amplification experiments were performed with QK1. The PCR reaction system is: the PCR reaction system is: 10X Buffer 2 μ l (containing Mg2+), dNTP0.4 μ l (10 mM), the nucleotide sequence shown in the sequence table SEQ ID NO: 3 and the sequence table SEQ ID NO: shown in 4 2 μl of each nucleotide sequence (5 μM), 1 μl of genomic DNA extracted from corn to be tested, 0.5 μl of taq enzyme (5 U / μl), the reaction volume is 20 μl, and a drop of mineral oil is added to cover. The PCR reaction program was: 94°C 5min; 94°C 60s, 53.5°C 60s, 72°C 60s, 35 cycles; 72°C 10min.

[0040] The PCR amplified products were analyzed by 6% polyacrylamide gel electrophoresis, s...

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Abstract

The invention discloses a molecular marker which is closely linked with corn bacterial wilt resistance genes. The molecular marker is a nucleotide sequence as shown in SEQ ID NO.1 in a sequence table or a nucleotide sequence as shown in SEQ ID NO.2 in the sequence table. A primer pair for amplifying the molecular marker is a nucleotide sequence as shown in SEQ ID NO.3 in the sequence table and a nucleotide sequence as shown in SEQ ID NO.4 in the sequence table. When the molecular marker is applied to corn bacterial wilt detection, amplification is performed through PCR, if an obtained PCR amplification product is a 234bp basic group fragment, the corn to be detected is of a bacterial wilt resistant variety, and if the obtained PCR amplification product is a 274bp alkali group fragment, the corn to be detected is of a bacterial wilt infected variety. By adopting the method, whether the corn has the bacterial wilt resistance or not can be detected before the corn is seeded, selective seeding can be achieved, the breeding time is shortened, and the method is rapid, simple and convenient and can be widely applied to assisted breeding of corn.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a method for detecting corn bacterial wilt resistance, PCR primers and applications thereof. Background technique [0002] Conventional breeding methods based on phenotypic selection have disadvantages such as low selection efficiency and long breeding cycle. It is urgent to inject modern molecular technology, supplemented by high-efficiency genotype-directed selection, in order to quickly and efficiently breed excellent new varieties of maize. With the rapid development of molecular biology and genomics, the application of molecular marker technology is more extensive. PCR-based molecular markers such as microsatellites or SSR (simple sequence repeat) are widely used because of their high polymorphism rate, relative stability, simple and fast detection method, and easy operation. Since molecular marker-assisted selection is not easily affected by environmen...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68A01H1/04
Inventor 朱先飞耿延琢王利明王兆贤张二朋杨焰华齐伟王丽梅
Owner HEFEI FENGLE SEED
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