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Molecular marker of rice seasonal febrile disease resistance gene Pigm and application of molecular marker

A molecular marker and blast resistance technology, applied in the field of molecular genetics, can solve the problems of low breeding efficiency and inaccurate disease resistance identification results, and achieve the effects of shortening the breeding cycle, high accuracy and improving efficiency.

Inactive Publication Date: 2017-08-11
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

In conventional disease resistance breeding, inaccurate identification of disease resistance results in low breeding efficiency

Method used

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  • Molecular marker of rice seasonal febrile disease resistance gene Pigm and application of molecular marker
  • Molecular marker of rice seasonal febrile disease resistance gene Pigm and application of molecular marker
  • Molecular marker of rice seasonal febrile disease resistance gene Pigm and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: the development of Pigm gene molecular marker

[0019] (1), the test material.

[0020] Disease-resistant rice variety: Gumei No. 4.

[0021] Susceptible rice varieties: 1: Hanhui 3, 2: BL6, 3: Huhan 1B, 4: B5, 5: SAGC4, 6: Nipponbare, 7: Huanghuazhan, 8: 603B, 9: 0412, 10: 389B, 11: R9, 12: Hua 15B, 13: R15, 14: Shanghai Han 9B, 15: R37, 16: Shanghai Han 11B, 17: N632S, 18: Xiushui 123, 19: R49, 20: Xiangqing, 21: Shanghai Drought 7B, 22:75-1-127.

[0022] (2) For the extraction method of rice genomic DNA, refer to the method described in Lou Qiaojun (2006). (Lou Qiaojun, Chen Liang, Luo Lijun. Comparison of three rapid extraction methods of rice genomic DNA[J]. Molecular Plant Breeding, 2006,3(5):749-752)

[0023] (3) Development of gene molecular marker gm-4: Pigm is a cloned broad-spectrum rice blast resistance gene, located on chromosome 6, and has an allelic relationship with Pi-9, Pi-z, Pi2, etc., Pigm The whole genome sequence of the genome is...

Embodiment 2

[0034] Example 2: Detection of parental polymorphisms of Pigm gene molecular markers

[0035] (1) The genome of the rice variety to be tested is extracted, and the extraction method is the conventional CTAB method. The rice varieties tested are: 1: Hanhui 3, 2: BL6, 3: Huhan 1B, 4: B5, 5: SAGC4, 6: Nipponbare, 7: Huanghuazhan, 8: 603B, 9: 0412, 10: 389B, 11: R9, 12: Hua 15B, 13: R15, 14: Huhan 9B, 15: R37, 16: Huhan 11B, 17: N632S, 18: Xiushui 123, 19: R49, 20: Xiangqing, 21 : Huhan 7B, 22: 75-1-127, 23: Gumei No. 4 Huhan 1B, Xiushui 123, Huhan 7B, Huang Huazhan, Zhongzu 14, Shanghai Han 11B, Nipponbare and 75-1-127.

[0036] (2), using the functional marker gm-4 to carry out PCR amplification on the tested rice variety, the PCR amplification system is: 20ng rice genomic DNA template, 10uL Taq PCR Mastermix (Tiangen Biochemical Technology (Beijing) Co., Ltd.), 10uM 1uL each of the front and rear primers, supplemented with dd H2O to 20uL. The PCR reaction program was: 95°C p...

Embodiment 3

[0038] Example 3: Verification and application of Pigm gene molecular markers

[0039] Using the disease-resistant parent Gumei 4 carrying the Pigm gene as the donor, and the water-saving and drought-resistant restorer line Hanhui 3 as the recipient, the rice blast resistance of Hanhui 3 was improved. A total of 384 individual plants of the BC1F2 population of Hanhui 3 / Gumei 4 / / Hanhui 3 were planted in the Natural Identification Base of Rice Blast in Jinggangshan. First, the genomic DNA of the 384 individual plants was paired with Pigm gene functional marker gm-4 PCR amplification was carried out, and the products were electrophoretically typed on 2% agarose gel, and 280 individuals were detected to have a 516bp band, indicating that these individuals contained the Pigm gene, and the remaining 104 individuals did not amplify 516bp Strips, conforming to a statistical 3:1 ratio. The results of rice blast resistance identification showed that all 280 individual plants containing...

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Abstract

The invention relates to a molecular marker of a rice seasonal febrile disease resistance gene Pigm and application of the molecular marker. The method for assisted-selection of seasonal febrile disease resistance rice by applying a molecular marker gm-4 comprises the following steps of: carrying out PCR (polymerase chain reaction) amplification by using DNA (deoxyribonucleic acid) of a rice genome as a template and gm-4 as a primer; and detecting an amplification product by using 2% agarose gel electrophoresis, if a 516bp stripe exists, a sample tested contains rice seasonal febrile disease resistance gene Pigm. The molecular marker has the beneficial effects that the genetic exchange does not exist through sequence difference design; and the molecular marker has high accuracy, is directly used for agarose gel electrophoresis detection, is relatively simple and convenient, is a co-dominant marker, can be used for detecting homozygous individuals and heterozygous individuals, shortens a breeding cycle and improves the efficiency.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a molecular marker of rice blast resistance gene Pigm and its application. Background technique [0002] The rice blast caused by Ascomycete [Magnaporthe grisea (Hebert) Barr] is one of the most serious diseases in rice production worldwide. The disease can occur in various growth stages of rice, and when it is severe, the yield can be reduced by 40%-50%. Harvest failure is one of the limiting factors for global food security. Extensive use of chemical pesticides pollutes the environment. The most economical and effective way to control this disease is to discover and utilize new broad-spectrum durable disease-resistant resources to breed new broad-spectrum disease-resistant varieties. [0003] Rice blast resistance is a typical quantitative trait controlled by multiple genes and susceptible to environmental influences. In conventional disease resistance breeding, inaccurate i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 刘毅刘国兰孔德艳唐金娟张安宁王飞名毕俊国余新桥罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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