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Molecular marker osblb-sv1 tightly linked to rice bacterial blight resistance gene

A technology of resistance to bacterial blight and molecular markers, applied in the field of molecular biology, can solve problems such as the complexity and lack of genetic mechanisms

Active Publication Date: 2019-03-01
SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, although some genes and quantitative trait loci (QTLs) related to rice bacterial blight resistance have been mapped, such as the broad-spectrum bacterial blight resistance Xa21 gene from Oryza longistaminata , the whole growth stage bacterial blight resistance Xa23 gene of common wild rice O. Molecular markers

Method used

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  • Molecular marker osblb-sv1 tightly linked to rice bacterial blight resistance gene
  • Molecular marker osblb-sv1 tightly linked to rice bacterial blight resistance gene
  • Molecular marker osblb-sv1 tightly linked to rice bacterial blight resistance gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: Construction of rice RIL population

[0040] Male parent: 801, resistant to bacterial blight, purchased from Shenzhen Huada Agriculture and Circular Economy Technology Co., Ltd.

[0041] Female parent: R15, not resistant to bacterial blight, purchased from Shenzhen Huada Agriculture and Circular Economy Technology Co., Ltd.

[0042] RIL population construction: The F1 generation was obtained by crossing the male parent and the female parent, and 188 recombinant inbred RIL strains (F7 generation) were obtained from the F1 generation by single seed transmission.

Embodiment 2

[0043] Embodiment 2: rice bacterial blight inoculation identification

[0044] A representative strain of dominant pathogenic type IV of bacterial blight of rice in Hunan Province (gifted by Hunan Hybrid Rice Research Center) was selected. Cultivate the strain with Wakimoto Tetsu's potato semi-synthetic medium at a constant temperature of 28°C for 72 hours, wash the bacterial lawn with sterile water, and dilute the bacterial suspension to 10 by McNerbidimetric method. 8 ~10 9 cells / ml of bacteria solution.

[0045] Wakimoto Tetsu’s potato semi-synthetic medium formula: 300 grams of potatoes, 15 grams of sucrose, 5 grams of peptone, 0.5 grams of Ca(NO3)2.4H2O, 2.0 grams of Na2HPO4.12H2O, 16 grams of agar powder, adjusted to 1000 ml with distilled water .

[0046] For the RIL population obtained in Example 1, the artificial leaf-cutting inoculation method was used to inoculate at the seedling stage of the plant, and 14 to 20 days after inoculation, the investigation was perfo...

Embodiment 3

[0049] Example 3: Extraction of Genomic DNA from Parents and RIL Individual Plants

[0050] The genomic DNA of the parents and 188 RIL individual plants were extracted by CTAB method, the specific method is as follows:

[0051] (1) Weigh 1.0g of fresh leaves, cut them into pieces and put them into a mortar, grind them with liquid nitrogen, add 3mL1.5×CTAB, grind them into a homogenate and transfer them to a 15mL centrifuge tube, then add 1mL1.5×CTAB into the mortar Wash with ×CTAB and transfer to a centrifuge tube. After mixing, place in a water bath at 65°C for 30 minutes, and shake slowly from time to time during this period.

[0052] Among them, the formula of 1.5×CTAB is as follows (1L):

[0053] CTAB

15g

1mol / L Tris.Cl (pH 8.0)

75mL

0.5mol / L of EDTA

30mL

NaCl

61.4g

[0054] Add deionized water to make up to 1 L, and add mercaptoethanol with a final concentration of 0.2% (2 ml) before use.

[0055] (2) After cooling to r...

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Abstract

The invention belongs to the field of molecular biology, relates to a molecular marker, and particularly relates to a molecular marker closely linked with genes of resistant bacterial blight of rice. The molecular marker contains the sequence shown in Seq ID No.1. The invention further relates to a primer for amplifying the molecular marker, the usage of the molecular marker and the primer in the positioning of the resistant bacterial blight of rice or the genetic breeding of rice, as well as a rice breeding method. The molecular marker provided by the invention links a genome DNA sequence with the genes of the resistant bacterial blight of rice, which is convenient for establishing a rice molecular marker assistant breeding system; the distance for genetic close linkage of the molecular marker and the genes of the resistant bacterial blight of rice is 0.2 cm. The molecular marker and the primer for amplifying the molecular marker can be simply and conveniently applied to rice breeding practice and resource and variety identification in a high throughput way.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to a molecular marker, in particular to a molecular marker closely linked with a rice bacterial blight resistance gene. The present invention also relates to the primer for amplifying the molecular marker, and the use of the molecular marker and the primer in rice bacterial blight resistance gene mapping or rice genetic breeding. Background technique [0002] Rice bacterial blight is a worldwide important disease. It is a disease of vascular bundles. Under natural conditions, pathogens usually invade through water holes or wounds and produce white spots along the veins. When bacterial blight occurs, it can lead to a 20% to 30% reduction in rice production, and even a complete failure in severe cases. In order to ensure high and stable production of rice, people mainly adopt two kinds of control techniques, one is to use chemical control methods, and the other is to cultivate and plan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/63C12Q1/6895
Inventor 张耕耘周承恕李宁刘晓娜刘小菊
Owner SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH
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