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Seed selection method for molecular marking assisted selection of high-resistance rice blast materials

A molecular marker-assisted, rice blast technology, applied in botany equipment and methods, plant gene improvement, application, etc., can solve the problems of heavy workload, dependence on disease resistance phenotype, low accuracy rate, etc., and achieve the effect of improving efficiency

Inactive Publication Date: 2014-12-03
国家粳稻工程技术研究中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional pedigree method selects the best breeding varieties from the separated offspring through the combination of two parents. The disease resistance phenotype depends on natural identification in the field or artificial inoculation identification, which is difficult to identify and has a low accuracy rate.
Because they do not know the disease-resistant genotype of the parents, the breeders are very blind in the selection of the parents, usually mass matching, massive selection of offspring, heavy workload and low breeding efficiency

Method used

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  • Seed selection method for molecular marking assisted selection of high-resistance rice blast materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: The resistant parent containing the rice blast resistance gene Pi40 was crossed with the susceptible parent Jijing 88 to construct a resistant and susceptible population and the identification and screening of the offspring population

[0016] The resistant parent NG13 containing the rice blast resistance gene Pi40 was crossed with the susceptible parent Jijing 88, and then selfed, and then the selfed F2 generation was screened for traits and the rice blast resistance gene Pi40, and the plant height, growth period, and high yield were selected. 50 individual F2 plants similar in sex to Jijing 88 and containing the disease resistance gene Pi40; then backcrossed with the susceptible parent Jijing 88 for the first time, and self-crossed to obtain BC 1 f 2 , and then screened for traits and disease-resistant gene Pi40, and selected BCs with excellent traits and disease-resistant gene Pi40 1 f 2 50 plants per plant; then carry out a second backcross with the sus...

Embodiment 2

[0017] Example 2: DNA extraction, PCR amplification and electrophoresis analysis of anti-susceptible individual plants

[0018] 1. DNA extraction (CTAB method)

[0019] Weigh 0.1~0.2g of rice leaves and cut them into 1cm long, put 800μl of extract solution (1M Tris PH8.0, 100mM: 5M Nacl, 1.0M EDTA, 20mM: 10%CTAB, 2.0%) preheated at 65℃ in a centrifuge tube. Put it into the tissue grinder, the frequency is 57.0HZ, the rotation speed is 1710N, and the vibration is 200S. Water bath at 65°C for 30 minutes, gently shake every 10 minutes evenly. Add an equal volume of phenol chloroform / isoamyl alcohol (25:24:1) and shake gently to form an emulsion. At room temperature (the temperature is not lower than 15°C), centrifuge at 12000r / min for 10min, and take the supernatant. Carefully extract 450 μl of the supernatant, add pre-cooled 2 times the volume of absolute ethanol (or an equal volume of isoamyl alcohol), place it at -20°C for more than 20 minutes, centrifuge at 5000 r / min for...

Embodiment 3

[0025] Example 3: The disease-resistant variety NG13 containing the rice blast resistance gene Pi40, the susceptible variety Jijing 88, and the resistant population were planted in Songyuan City, Jilin Province, a high-incidence area for rice blast, and the rice blast was induced by natural conditions. The investigation of the resistance to leaf blast and panicle blast was carried out at the stage and mature stage respectively, and the unified standard was used for evaluation.

[0026] 1. Field identification of leaf blast

[0027] Standards for investigation and recording of leaf blast:

[0028] Level 0 disease-free………………………………………………………………………………………… High resistance (HR)

[0029] Grade 1 lesions are needle-shaped brown spots……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………

[0030] Level 2 Slightly bigger brown spot………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………Resistance (R...

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Abstract

The invention relates to a seed selection method for molecular marking assisted selection of high-resistance rice blast materials, and belongs to the field of rice breeding. Specific molecular marker is assisted to select the diseases resistance gene materials, and field diseases resistance identification is combined, so the seed selection target is clear and efficient. The method includes the following steps: 1, hybridizing a resistant parent containing rice blast resistant gene Pi40 with a susceptible parent Jijing 88 to construct an anti-infectious group, and identifying and screening the above progeny group; 2, extracting DNA of an individual anti-infectious strain, carrying out PCR amplification, and carrying out electrophoresis analysis; and 3, planting rice blast resistance gene Pi40-containng rice variety NG13 and susceptible variety Jijing 88 and the individual anti-infectious group progeny strain in a rice blast high incidence area Songyuan city in Jilin Province, inducing the rice blast under natural conditions, and respectively carrying out leaf and panicle blast resistance investigation in the seedling and mature stage. The field anti-infectious group is constructed, a molecular marker for screening the rice blast resistant gene Pi40 is used, and the disease resistance identification of a rice blast high incidence area is combined, so efficient directed improvement and cultivation of rice blast resistant new varieties are realized, and the method is of great practical significance for field breeding.

Description

technical field [0001] The invention relates to a molecular marker-assisted selection method for rice blast-resistant materials, in particular, specific molecular marker-assisted selection of rice materials containing rice blast resistance genes, combined with field phenotypic traits and disease resistance identification, The breeding target is clearer and more efficient, and belongs to the field of rice breeding. Background technique [0002] Rice blast is a disease of rice caused by Magnaporthe grisea, and its distribution range is very wide. According to statistics, it occurs in more than 80 countries in the world, and it can cause a 10%-20% reduction in production in general epidemic years, and it can reach 40%-50% in severe cases. Since the 1990s, the annual occurrence area of ​​rice blast in my country has been 3.8 million ha 2 Above, hundreds of millions of kilograms of rice are lost every year. With the simplification and centralization of varieties and the impact...

Claims

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Application Information

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IPC IPC(8): A01H1/02A01H1/04C12Q1/68
Inventor 刘欣亓娜何广生朱崴华泽田
Owner 国家粳稻工程技术研究中心
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