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Rice cold-resistant major gene identification method and special primer thereof

A major gene and identification method technology, applied in the field of biomolecular markers, can solve problems such as large errors and easy occurrence of false positives, and achieve the effects of high cost, shortened breeding cycle, and clear selection goals.

Active Publication Date: 2014-06-18
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the problems of large errors in the existing detection and identification of cold-tolerant rice, which are prone to false positives, etc., and to provide a main gene for cold-resistant rice at seedling stage qCTS-3-1 The special primers for the molecular markers, and the method for identifying and screening cold-tolerant rice using the special primers

Method used

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  • Rice cold-resistant major gene identification method and special primer thereof
  • Rice cold-resistant major gene identification method and special primer thereof
  • Rice cold-resistant major gene identification method and special primer thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: rice hybridization and molecular marker analysis

[0036] (1) Construction of chromosome substitution lines

[0037] The cold-tolerant donor parent is the Guangxi common wild rice core germplasm material DP30, and the recipient parent is the sequenced indica variety 9311. Through backcrossing and SSR marker-assisted selection, a chromosome segment substitution line of common wild rice was established. F 1 Continuous backcross with 9311 to BC 2 f 1 , in BC 2 f 1 Begin to use 325 polymorphic SSR markers to do genotype analysis on 10 individual plants of each line, to BC 4 f 1 226 lines containing DP30 fragments were obtained, and 96 pairs of markers evenly distributed on the 12 rice chromosomes were selected for background screening, and the individual plants with a recovery rate of more than 90% were selfed and backcrossed with 9311 to obtain shorter Import the substitution system for the segment. Analyze BC 4 f 1 genotypes, self-crossed indiv...

Embodiment 2

[0045] Example 2: Identification of cold tolerance of parents and targeted populations

[0046] The cold tolerance of the donor parent DP30, the cold-tolerant control Fujisaka 5 and the recipient parent 9311 were identified at the seedling stage. The rate of viable seedlings of 9311 as the recipient parent, the perceptual control, was 6.7%. The rates of viable seedlings of the two donor parents DP30 and the cold-tolerant control Fujisaka No. 5 were 85.7% and 88.5%, respectively. The results are shown in Table 1.

[0047] Table 1 Cold tolerance of parents and controls

[0048]

[0049] Using the established 230 BCs covering the whole genome 4 f 2 The cold tolerance level (live seedling rate) of each family was continuously distributed, and the cold tolerance of the DC907 family was the strongest at 83%, while the lowest was 0. The DC907 family with strong resistance was selected to construct a QTLs marker mapping population, and 332 individual plants were identified f...

Embodiment 3

[0057] Example 3 Verification of molecular markers

[0058] Negative varieties of materials used: 80 cold-intolerant materials in the breeding combination of cold-intolerant variety 9311 and DP30×9311.

[0059] Positive varieties: 19 cold-tolerant materials in the breeding combination of cold-tolerant parents wild rice DP30 and DP30×9311. Molecular marker primers: RM15040 , ZCT13 , ZCT23 , RM15123 .

[0060] Verification method 1: CTAB extraction method to extract positive rice (with cold tolerance main gene qSCT-3-1 rice), negative rice (does not have cold tolerance main gene qSCT-3-1 rice) sample genomic DNA (the method is the same as in Example 1), and the sample DNA was amplified with labeled primer RM15040. Forward primer (SEQ ID NO:1) 0.10μM, reverse primer (SEQ ID NO:2) 0.10uM, 250μM dNTPs, 1.0ul of 10×PCR reaction buffer (50mM KCl, 10mM Tris-HCl pH8.3 , 1.5mM MgCl 2 ), rice genomic DNA template 20ng, Taq DNA polymerase 1U, supplement the reaction system ...

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Abstract

The invention relates to a rice seedling stage cold-resistant major gene and a special primer thereof. The rice cold-resistant major gene for the special primer comprises the following steps: by taking a rice genome DNA to be detected as a template, performing polymerase chain reaction (PCR) amplification by using any one labeled primer in a labeled primer RM15040, a labeled primer ZCT13, a labeled primer ZCT23 and a labeled primer RM15123, and performing gel electrophoresis assay on the PCR amplification product to obtain rice cold-resistant identification, wherein stripes with the size of 140-170bp or 230-270bp or 290-310bp or 380-400bp exist in the PCR amplification product, and the 3rd chromosome of the seedling stage rice contains cold-resistant major genes qCTS-3-1. According to the rice cold-resistant screening, the production cost can be saved, the selection efficiency is improved, the breeding cycle of rice varieties is shortened, and the errors of the detection result are reduced.

Description

technical field [0001] The invention belongs to the field of biomolecular markers, and in particular relates to a molecular marker of rice seedling stage cold-tolerant main gene qCTS-3-1, and a method for identifying and screening cold-tolerant rice using special primers. Background technique [0002] Low temperature and chilling injury of rice has occurred in many countries in the world, and it is a global natural disaster. In the temperate zone where rice is widely planted, as well as in tropical and subtropical regions, chilling injury occurs frequently, seriously affecting the stability and development of rice production, and has also had a considerable impact on the security and coordination of world food production. Chilling damage occurs in all rice areas in my country, and a major chilling damage occurs every 4 to 5 years, causing the annual loss of 5 billion to 10 billion kg of rice in my country in a disaster year. After sowing early rice in the double-croppin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113
Inventor 李容柏郑加兴陈保善邱永福刘芳覃宝祥蒙姣荣
Owner GUANGXI UNIV
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