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Primers and kit for detecting rice blast resistance genes Pigm and genetic typing method

A rice blast resistance gene and rice blast resistance technology, applied in the field of kits and genotyping, detection primers of rice blast resistance gene Pigm, can solve problems such as environmental pollution, misjudgment of test results, human hazards, etc., and achieve sensitivity High, strong specificity, the effect of eliminating aerosol pollution

Inactive Publication Date: 2016-04-20
YUAN LONGPING HIGH TECH AGRI CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the molecular markers used to detect this gene are dominant or non-closely linked markers, and the detection results are likely to cause misjudgment; the EB or polyacrylamide used in the detection process is likely to pollute the environment and cause harm to the human body

Method used

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  • Primers and kit for detecting rice blast resistance genes Pigm and genetic typing method
  • Primers and kit for detecting rice blast resistance genes Pigm and genetic typing method
  • Primers and kit for detecting rice blast resistance genes Pigm and genetic typing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] This embodiment is intended to illustrate the screening process of primers and probes of the present invention

[0035] 1. Experimental materials and sources

[0036] A total of 10 rice materials were selected for testing, specifically: Gumei 4, 9311, R1128, Huanghuazhan, Zhongguangsimiao, R608, Huazhan, and Gumei 4 as the donor and 9311 as the recurrent parent. Near-isogenic lines B170, B195, Z2752 (BC 6 f 2 single plant). Among them, Gumei 4, B170, and B195 were resistant to blast strain 67, and Z2752, 9311, R1128, Huanghuazhan, Zhongguangsimiao, R608, and Huazhan were susceptible. The above materials were provided by Hunan Yahua Seed Industry Science Research Institute.

[0037] 2. DNA extraction

[0038] For 10 test materials, a few leaves were taken at the seedling stage to extract genomic DNA by CTAB method. NaroDrop2000 micro-measure DNA concentration, and adjust the concentration to about 50ng / μl.

[0039] 3. Sequencing and sequence comparison

[0040] A...

Embodiment 2

[0043] Example 2 A Taqman qualitative genotyping PCR primer for detecting rice blast resistance gene Pigm

[0044] The purpose of this embodiment is to provide a kind of Taqman qualitative genotype analysis PCR primers for detecting rice blast resistance gene Pigm, said primers comprising:

[0045] Upstream primer S1806GC-F: 5'-ACTCCTTTCATCCCATAAAATACAAACGT-3';

[0046] Downstream primer S1806GC-R: 5'-CGCCTCTCCAGATTTGCAGTAT-3'. Example 3 The probe used in conjunction with the primers described in Example 2

Embodiment 3

[0046] Downstream primer S1806GC-R: 5'-CGCCTCTCCAGATTTGCAGTAT-3'. Example 3 The probe used in conjunction with the primers described in Example 2

[0047] This embodiment aims to provide a probe used in conjunction with the primers described in Example 2, the probe comprising:

[0048] S1806-VIC: 5'-CCCAAATGTTAGAGATCATA-3';

[0049] S1806-FAM: 5'-CCCAAATGTTAGACATCATA-3'.

[0050] Among them, VIC and FAM are fluorescent reporter groups.

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Abstract

The invention relates to the bio-detecting technology, in particular to a primer and a kit for detecting rice blast resistance genes Pigm and a genetic typing method. By means of the method of comparing alleles of multiple rice blast resistance genes Pigm with a 'GuMei 4' near-isogenic line sequence, a basic group difference site capable of being different from other susceptible alleles is obtained through analysis, and then design screening is carried out, so that the optimal primers S1806GC-F and S1806GC-R and optical probes S1806-VIC and S1806-FAM are obtained. The invention further discloses a genetic typing method for the rice blast resistance genes Pigm through the primers and the probes. Genetic typing analysis is carried out through the fluorescence real-time PCR technology, enzyme digestion, electrophoresis and sequencing are not needed, operation is easy and convenient, time and labor are saved, and safety and environmental friendliness are achieved.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a primer, a kit and a genotyping method for detecting rice blast resistance gene Pigm. Background technique [0002] Rice blast caused by Magnapotheoryzae is one of the most serious diseases to rice production, which can cause 10-30% yield reduction every year. Breeding and utilizing disease-resistant varieties is the safest and most effective way to control rice blast. The rice variety "Gumei No. 4" has been identified as a highly resistant variety by the Hunan Rice Blast Disease Nursery for many years. With "Gumei No. 4" as the donor, the rice variety selected and promoted has been widely used in commercial breeding, and its resistance to rice blast is stable and high. [0003] The Pigm gene carried by "Gumei No. 4" is one of the genes with broad resistance spectrum and long-lasting resistance. The Pigm gene was located in the segment of chromosome 6 10367284-10421563 of th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 曾晓珊杨远柱白珍安秦鹏彭丹胡小淳
Owner YUAN LONGPING HIGH TECH AGRI CO LTD
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