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Molecular marker sv3 closely 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 the problems of lack of QTLs and molecular markers, complex genetic mechanisms, etc.

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

[0007] At present, although some genes and QTLs related to rice bacterial blight resistance have been mapped, due to the complex genetic mechanism of rice bacterial blight resistance, there is still a lack of sufficient numbers of QTLs and molecules related to bacterial blight resistance. mark

Method used

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  • Molecular marker sv3 closely linked to rice bacterial blight resistance gene
  • Molecular marker sv3 closely linked to rice bacterial blight resistance gene
  • Molecular marker sv3 closely linked to rice bacterial blight resistance gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Construction of rice RIL population

[0039] Rice seeds were purchased from: Shenzhen Huada Agriculture and Circular Economy Technology Co., Ltd.

[0040] Male parent: 801, resistant to bacterial blight;

[0041] Female parent: R15, not resistant to bacterial blight;

[0042] The F1 generation is obtained by crossing the male parent and the female parent;

[0043] 188 recombinant inbred lines RIL populations (F7 generation) were obtained from the F1 generation by the method of single-grain subculture.

Embodiment 2

[0044] Embodiment 2: rice bacterial blight inoculation identification

[0045] 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. The bacterial strain was cultured with Wakimoto's potato semi-synthetic medium at a constant temperature of 28°C for 72 hours, the bacterial lawn was washed with sterile water, and the bacterial suspension was diluted to 108-109 cells / ml bacterial solution by McNairney turbidimetry. Medium formula: 300 grams of potato, 15 grams of sucrose, 5 grams of peptone, Ca(NO 3 ) 2 .4H 2 O 0.5 g, Na 2 HPO 4 .12H 2 O 2.0 grams, agar powder 16 grams, distilled water adjusted to 1000 ml.

[0046] For the RIL population obtained in Example 1, the artificial leaf-cut inoculation method was used to inoculate at the seedling stage of the plant, and 14 to 20 days after the inoculation, when the condition of the susceptible variety was stabilized, the inv...

Embodiment 3

[0050] Example 3: Extraction of Genomic DNA

[0051] Genomic DNA of individual plants of the male parent, female parent, F1 generation and RIL population were extracted by the CTAB method, and the specific methods were as follows:

[0052] (1) Weigh 1.0g of fresh leaves, cut them into pieces and put them in a mortar, grind them with liquid nitrogen, add 3mL 1.5×CTAB, grind them into a homogenate and transfer them to a 15mL centrifuge tube, then add 1mL 1.5×CTAB into the mortar Rinse 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.

[0053] Among them, the formula of 1.5×CTAB is as follows (1L):

[0054] CTAB

15g

1mol / L Tris.Cl (pH is

75mL

[0055] 8.0)

0.5mol / L of EDTA

30mL

NaCl

61.4g

[0056] Add deionized water to make up to 1 L, and add mercaptoethanol with a final concentration of 0.2% (2 ml) before use. ...

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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 0.5 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 (Oryza sativa) is one of the most important food crops in my country, and rice is the staple food of more than 60% of the population in my country. Therefore, the improvement of rice production level has always been a major issue related to the development of agriculture and the stability of people's livelihood in our country. [0003] 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895C12N15/70C12N1/21
Inventor 倪雪梅蔡雪梅雷雪静张荣黄聪
Owner SHENZHEN BGI AGRI & CYCLE ECONOMIC TECH
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