Molecular marker primer of rice gall midge-resistant major gene Gm5 and marking method and application of molecular marker primer

A major gene and molecular marker technology, applied in the field of molecular genetics, can solve the problems of difficult introduction and aggregation of insect-resistant genes, complex identification of insect-resistance, etc., and achieve the effects of convenient identification, convenient and rapid detection, and improved screening efficiency.

Active Publication Date: 2020-03-27
GUANGXI UNIV
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the complexity of insect resistance identification, it is often difficult to effectively introduce and aggregate different insect resistance genes using conventional breeding methods

Method used

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  • Molecular marker primer of rice gall midge-resistant major gene Gm5 and marking method and application of molecular marker primer
  • Molecular marker primer of rice gall midge-resistant major gene Gm5 and marking method and application of molecular marker primer
  • Molecular marker primer of rice gall midge-resistant major gene Gm5 and marking method and application of molecular marker primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Acquisition of molecular markers

[0032] 1. F2 population construction and phenotypic identification

[0033] (1) Indica rice variety ARC5984 is highly resistant to rice gall midge Indian biotypes 1, 2 and 5 (Kumar et al.1998), and ARC5984 is highly resistant to Chinese biotypes 2 and 4 (Wu et al.2014). This insect-resistant variety comes from the Plant Protection Research Institute of Guangxi Academy of Agricultural Sciences. F obtained after 9311 / ARC5984 hybridization experiment 1 The individual re-selfed to obtain F 2 offspring, each F 2 A single plant obtains the corresponding F by selfing 2：3 Families were screened for insect resistance. Insect resistance identification experiments showed that ARC5984 had high resistance to rice gall midge populations from Nanning rice fields;

[0034] (2) Adopt seedling stage inoculation treatment to parents, F 2:3 Families were tested for insect resistance. To ensure that the parent and F 2:3 Each family in the group grow...

Embodiment 2

[0048] Validation of Molecular Markers

[0049] 1. Materials and Methods

[0050] (1) Material

[0051] Negative varieties: susceptible lines (species) 9311 and MH63, both of which are conventional rice materials preserved in our laboratory.

[0052] Positive varieties: high insect-resistant strains (species) ARC5984, 570011 and ARC5833, all of which are conventional rice materials preserved in our laboratory.

[0053] There are 28 offspring of ARC5984×9311 hybrid combination.

[0054] Molecular marker primer: Z22595

[0055] (2) method

[0056] The genomic DNA of rice sample leaf was extracted by CTAB extraction method, the sample DNA was amplified with primer Z22595, and the amplified product was separated by 3% agarose gel electrophoresis (the method is the same as in Example 1).

[0057] 2. Results:

[0058] Using the method above, PCR amplification was performed on the genomic DNA of 33 different samples of rice lines 570011, 9311, and ARC5984, respectively. Such a...

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Abstract

The invention relates to the field of molecular genetics, in particular to a molecular marker primer of a rice gall midge-resistant major gene Gm5 and a marking method and application of the molecularmarker primer. According to the method, the genotype of each single plant of F2 obtained by hybridizing a rice insect-resistant variety ARC5984 (male) and 9311 (female) is acquired, meanwhile, genetic linkage analysis is conducted by combining the rice gall midge-resistant resistance level of an F<2:3> family, the rice gall midge-resistant major gene Gm5 carried by the insect-resistant variety ARC5984 is identified, the region of the Gm5 is reduced to a 49kb fragment defined by markers Z57 and Z64 on the two sides, and the region is closely linked with the molecular marker Z22595. The molecular marker Z22595 can effectively detect whether the insect-resistant variety ARC5984 and derived varieties (lines) thereof contain a major resistance gene locus or not, the selection efficiency of rice gall midge-resistant rice plants is greatly improved, and the rice gall midge-resistant rice varieties (lines) containing the Gm5 are obtained.

Description

【Technical field】 [0001] The invention belongs to the field of molecular genetics, and in particular relates to a molecular marker primer of the rice gall midge-resistant main gene Gm5, a marker method and application thereof. 【Background technique】 [0002] Rice is one of the most important agricultural crops in the world and its production is threatened by various pathogens, pests and other harmful organisms. The rice gall midge (GM, Orseolia oryzae, Diptera: Cecidomyiidae) feeds on the sap of rice growth points in the larval stage, so that the heart leaves stop growing after the growth points are damaged, and the leaf sheaths elongate into tubes, that is, "standard onions" appear, resulting in the inability of rice heading. According to statistics, the damage of rice gall midge alone can cause an annual yield loss of more than 550 million US dollars (Herdt 1991). Chemical pesticides commonly used for rice gall midge control can lead to environmental pollution or the eme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156Y02A50/30
Inventor 邱永福周海连王心怡莫怿
Owner GUANGXI UNIV
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