Rice blast resistance gene Piz-t functional specific molecular marker and detection method and application thereof

A resistance gene and molecular marker technology, applied in the field of agricultural biology, can solve the problems of easy misselection or missed selection in resistance breeding, and achieve the effects of low cost, high throughput and high accuracy

Inactive Publication Date: 2017-05-24
HUAIYIN TEACHERS COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Huaxia et al. (2015) developed a dominant marker of Piz-t, which is prone to misselection or missed selection in resistance breeding.

Method used

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  • Rice blast resistance gene Piz-t functional specific molecular marker and detection method and application thereof
  • Rice blast resistance gene Piz-t functional specific molecular marker and detection method and application thereof
  • Rice blast resistance gene Piz-t functional specific molecular marker and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Rice blast resistance gene Piz-t gene-specific molecular marker and its primer design and detection:

[0034] (1) Analysis of Piz-t insertion / deletion (insertion-deletion, InDel) site:

[0035] Partial genome sequences of rice donor varieties such as Piz-t were downloaded from public databases, and for NBS 1～9 -Piz-t site for sequence comparison, screening Piz-t specific, specific insertion / deletion (InDel) differential site design primers, amplified IRBLz-FU(Piz), Gumei 2(P26(t)), Gumei 4(Pigm(t)), Erbazhan(Pi50(t)), IR65482-4-136-2-2(Pi40(t) )), Nipponbare, the sequencing and comparison results are shown in the table below:

[0036]

[0037] Wherein, ---- is the identified Piz-t gene-specific deletion sequence;

[0038] (2) Design primers:

[0039] According to the design principle of InDel labeling, primers are designed at the upstream and downstream 100-200bp of the InDel site, and the primer pair base sequences are as follows:

[0040] Fl: 5'-TTACTAGAATCGCT...

Embodiment 2

[0057] Detection application of resistance gene Piz-t gene-specific molecular markers in other rice varieties:

[0058] Select 13 representative rice varieties, in order: IRBLzt-T (Piz-t), Aiyangdao, Liaojing 5, TQ155, Masoli, BG1100, TRAT, 5011, Peiai 64, 9311, Yanghui 136-4 , Zhongnongqing, 34 Chongqing Hui.

[0059] Genomic DNA was extracted respectively, and using this as a template, PCR amplification, enzyme digestion and electrophoresis detection were performed according to the method of Example 1. According to the size of the digested products (bands), the resistance gene Piz-t can be distinguished from other rice blast resistance genes. Such as figure 2 As shown, the Piz-t-InDel gene specificity can be detected in individuals whose resistance spectrum phenotype is Piz-t type (the disease-resistant variety IRBLZT-T carrying the rice blast resistance gene Piz-t, the sample of lane 1). The presence of sex molecular markers, while individuals with other spectrum-resist...

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Abstract

The invention provides a rice blast resistance gene Piz-t functional specific molecular marker and a detection method and application thereof. The molecular marker shows a specific banding pattern of the rice blast resistance gene Piz-t amplified from a rice genome DNA through primer pairs SEQ ID NO.1 and the SEQ ID NO.2. The rice blast resistance gene Piz-t gene specific molecular maker provided by the invention has important application values, and the utilization efficiency of the gene in each of germplasm resource screening, molecular marker-assisted selective breeding, gene pyramiding breeding and transgenic breeding can be improved by use of the marker.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a rice blast resistance gene Piz-t functional molecular marker Piz-t-InDel and a detection method and application thereof. Background technique [0002] Rice blast is one of the most serious diseases that endanger the high and stable yield of rice. Although the chemical control commonly used in current production can solve or reduce the impact of diseases on rice production to a certain extent, long-term use of pesticides in farmland not only pollutes the environment, but also affects the edible quality of rice due to pesticide residues. Long-term production practice has shown that breeding and utilizing disease-resistant varieties is one of the safest and most effective methods for controlling rice blast. However, due to the frequent variation of the pathogenicity of the physiological races of Magnaporthe grisea, this will lead to the gradual loss of resist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 杨立明纪剑辉周颖君罗玉明
Owner HUAIYIN TEACHERS COLLEGE
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