Molecular marker for rice brown planthopper-resistance QBph3 and QBph4 genes

A technology of anti-BPH and molecular markers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems such as the difficulty of effectively introducing or aggregating multiple BPH-resistant gene insect-resistant varieties, etc., to achieve Improve the efficiency of assisted selection, strong operability, and high primer specificity

Inactive Publication Date: 2015-11-25
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the complexity of insect resistance identification, it is often difficult to effectively introduce...

Method used

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  • Molecular marker for rice brown planthopper-resistance QBph3 and QBph4 genes
  • Molecular marker for rice brown planthopper-resistance QBph3 and QBph4 genes
  • Molecular marker for rice brown planthopper-resistance QBph3 and QBph4 genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1: Preliminary localization of QBph3 and QBph4 in the insect-resistant introduction line IR02W101

[0075] 1. Build Zhenshan 97 / IR02W101BC 1 f 2 Population and Phenotype Identification

[0076] (1) The insect-resistant parent rice line IR02W101 (formerly known as IR54751-1-2-44-15-2-B) was bred by the Early International Rice Research Institute ("IRRI" in English, Philippines), and was developed by the Kindly provided by Dr. Brar of IRRI. They crossed a conventional indica rice variety IR31917-45-3-2 with a medicinal wild rice variety (Acc number: 100896), and after phenotypic identification and backcross breeding, they finally obtained the introduction of four highly resistant brown planthopper biotypes Line IR02W101 (see literature: Jena and Khush, 1990; Brarand Khush, 1997). The other parent Zhenshan 97 (Zhenshan 97B) used in this example showed high sensitivity to the brown planthopper population in my country. In order to locate the brown planthopper res...

Embodiment 2

[0089] Example 2: Fine Mapping of QBph3 and QBph4

[0090] 1. Fine positioning of QBph3

[0091] (1) In order to further finely locate QBph3 and improve selection efficiency, select BC 1 f 3 The QBph3 locus in the family is the IR02W101 genotype, and the QBph4 locus is the single plant of the Zhenshan 97 genotype, which was further backcrossed with Zhenshan 97 to obtain BC 2 f 1 , and then use the QBph3 flanking markers RM514 and J412 to identify BC 2 f 1 genotypes, select the individual plants that are heterozygous for the two marker genotypes for further selfing, and obtain BC 2 f 2 group. Then mark RM514 and J412 on both sides of the QBph3 site to detect a total of 6000 BC 2 f 2 In the family, a total of 220 recombined individuals with crossover between markers on both sides were found. And for each recombinant individual plant corresponding F 2:3 The family is identified for insect resistance, and its resistance score is obtained, and then the genotype of the in...

Embodiment 3

[0103] Example 3: Gene effect analysis of QBph3 and QBph4 in Zhenshan 97 background

[0104] 1. Material construction and resistance identification

[0105] According to the method of molecular marker-assisted selection (MAS) combined with conventional backcross breeding, the two main anti-BPH genes QBph3 and QBph4 in IR02W101 were simultaneously introduced into the susceptible parent Zhenshan 97. First select the BC of Zhenshan 97 / IR02W101 2 f 3 A single plant of the IR02W101 genotype at two gene loci in the family was further backcrossed with Zhenshan 97 to obtain BC 3 f 1 , using the markers c3-14 and xc4-27 closely linked to QBph3 and QBph4 to identify the genotype of the individual plants, and screened out the individual plants whose marker genotypes were heterozygous at the QBph3 and QBph4 loci and further selfed to obtain BC 3 f 2 Segregated populations, at this time, the two genes QBph3 and QBph4 should be segregated at the same time, and the genotypes of the offs...

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Abstract

The invention belongs to the technical field of preparation of a rice molecular marker, and in particular relates to a molecular marker for two rice brown planthopper-resistance major genes QBph3 and QBph4 which are simultaneously derived from a pest-resistance introgression line IR02W101. The molecular marker comprises the following steps: carrying out crossing, self-crossing and backcrossing on Zhenshan97, which is an herbivore-susceptible variety, and IR02W101 so as to obtain genotypes of various BC1F2 single plants; carrying out genetic linkage analysis in accordance with brown planthopper-resistance grades of various F2: 3 lines during seedling stage; precisely locating a resistance gene QBph3 of the IR02W101 between long-arm markers t6 and f3 of the 3rd chromosome so as to obtain a co-segregative marker c3-14 as well as closely linked markers q1 and m3; and precisely locating QBph4 between short-arm markers p17 and xc4-27 of the 4th chromosome so as to obtain closely linked markers p6, p9, c4-5, xc4-7, HJ16, J417 and IN156. The molecular marker disclosed by the invention can be used for effectively detecting whether the pest-resistance introgression line IR02W101 and derived varieties thereof contain the major gene site or not.

Description

technical field [0001] The invention belongs to the technical field of plant molecular marker preparation. It specifically relates to the preparation of molecular markers derived from the tight linkage of two brown planthopper resistance genes QBph3 and QBph4 in the introduced line IR02W101, and the application of the molecular markers in rice brown planthopper resistance breeding and variety improvement. Background technique [0002] Rice is the main food crop in my country, and the brown planthopper is currently the main pest active in the rice region of Southeast Asia. It is a typical piercing-sucking pest. It sucks the phloem juice of rice plants through its stylet. If it is mild, it will cause the transportation of rice nutrients to be blocked and affect the thousand-grain weight. Lice burn", leading to a large area of ​​rice production reduction or even failure. At present, the control of brown planthopper mainly relies on chemical insecticides, but this method is ti...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 何予卿胡杰高冠军张庆路
Owner HUAZHONG AGRI UNIV
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