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Method for molecular marker assisted selection of rice material with high rice blast resistance

A rice blast and high-resistance technology, which is applied in botany equipment and methods, biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of difficult identification, blindness, and heavy workload

Inactive Publication Date: 2020-07-07
国家粳稻工程技术研究中心
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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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  • Method for molecular marker assisted selection of rice material with high rice blast resistance

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 spots………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………

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Abstract

The invention relates to a method for molecular marker assisted selection of a rice material with high rice blast resistance. The method adopts specific molecular markers to assist in selection of a disease-resistant gene material, combines identification of field disease resistance to achieve a clearer and more efficient breeding target, and belongs to the field of rice breeding. The method comprises the following steps: (1) hybridizing a resistance parent containing a rice blast resistance gene Pi40 and an susceptible parent Jijing 88 to construct an anti-susceptible group, and identifying and screening a progeny group; 2) extracting DNA of an anti-infection single plant, and performing PCR amplification and electrophoretic analysis; and 3) planting the rice variety NG13 containing the rice blast resistance gene Pi40, the susceptible variety Jijing 88 and the anti-susceptible group progeny single plant in the rice blast high-incidence area of Songyuan city in Jilin Province, inducingrice blast under natural conditions, and respectively carrying out resistance investigation on leaf blast and spike-stem blast in a seedling stage and a mature stage. According to the invention, through construction of a field anti-infection group, utilization of the molecular markers for screening the rice blast resistance gene Pi40, and combination of disease resistance identification of a riceblast high-incidence area, a novel rice-blast-resistant variety can be efficiently and directionally improved and cultivated, and the method has important practical significance on 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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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H1/04C12Q1/6895
CPCA01H1/02A01H1/04C12Q1/6895C12Q2600/13
Inventor 高星杨坤姜丽杰
Owner 国家粳稻工程技术研究中心
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