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Molecular marking method of peanut plant type-related gene loci and application thereof

A gene locus and molecular marker technology, applied in the field of molecular genetics and molecular breeding, to achieve the effect of cost saving, quick and easy identification method and high selection efficiency

Inactive Publication Date: 2019-12-20
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The existing research is limited to this, and there is no systematic and molecular genetic research on the peanut plant type represented by the peanut side branch angle.

Method used

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  • Molecular marking method of peanut plant type-related gene loci and application thereof
  • Molecular marking method of peanut plant type-related gene loci and application thereof
  • Molecular marking method of peanut plant type-related gene loci and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Genetic Mapping of Peanut Side Branch Angle, Growth Habit and Plant Type-Related Gene Loci

[0035] (1) Materials and methods:

[0036] (1) Construction of plant materials and genetic populations: the creeping cultivated peanut Xiaohongmao was crossed with the erect cultivated peanut Henan Nanyang peanut, the hybrids were planted, and F 2 Separation of populations, continuous self-breeding of multiple generations of strains, and finally the formation of F 6 Recombinant inbred line HN-F 6 . It is used for the detection and location of the angle-related gene loci between the main effect and side branches.

[0037] (2) Extract individual DNA by CTAB method (refer to "Molecular Cloning Experiment Guide" for details).

[0038] (3) Development of polymorphic InDel markers: According to the comparison between the resequencing data of the parental genome and the reference genome (Tifrunner genome data: https: / / www.peanutbase.org / peanut_genome), 31 pairs of InDel ...

Embodiment 2

[0049] (1) Development and application of co-segregation marker LBA5b-caps for lateral branch angle, growth habit and plant type related gene loci

[0050] According to the result of embodiment 1, the difference of TA insertion between parents on the candidate gene LBA5b is developed as a CAPS marker, which is named as LBA5b-caps (see Figure 4 ). Amplify HN-F using the upstream and downstream primers of CAPS-labeled LBA5b-caps 6 The genomic DNA of each individual in the population is digested by restriction endonuclease SacⅠ, and then detected by electrophoresis on 1% agarose gel electrophoresis. If the PCR product is cut into two small fragments, it means that the individual has an LBA5b potentiation allele If the PCR product cannot be cut at the mutated gene locus (from Xiaohongmao), it means that the individual does not have the LBA5b synergistic variant gene, but has a depleted allelic variant lba5b (from Nanyang peanut). Allelic variation analysis and phenotypic analysi...

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Abstract

The invention belongs to the field of molecular genetic breeding science, and provides a molecular marking method of gene loci related to peanut lateral branch angle, growth habit and plant types andapplication thereof. By designing a CAPS molecular marker, and by using a method combining primer amplification, enzyme digestion and agarose gel electrophoresis, the judgment of allelic variation ofgene loci related to the lateral branch angle, the growth habit and the plant types is realized. By labeling PM15-ID1 and PM15-ID2 with InDel, and by using the method of primer amplification and polyacrylamide gel electrophoresis, the marker-assisted selection of offspring with allelic variation of gene loci related to lateral branch angle, growth habit and plant types. The molecular marking method provided by the invention can be used for detecting the offspring of a breeding group constructed by using peanut creeping germplasm and upright germplasm as parents, can be used for rapidly identifying the allelic variation of gene loci related to lateral branch angle, growth habit and plant types of the offspring, can improve the selection efficiency of the gene loci, and can provide referencefor peanut plant type improvement and high-yield breeding.

Description

Technical field: [0001] The invention belongs to the technical field of molecular genetics and molecular breeding, and relates to a molecular marker method for peanut lateral branch angle, growth habit and plant type-related genes, which is used for molecular marker-assisted improvement of cultivated peanut lateral branch angle and molecular marker-assisted peanut plant type breeding and Utilization of germplasm resources. Background technique: [0002] Peanuts (Arachis hypogaea L.2n=4x=40, AABB allotetraploid) are rich in edible oil (about 38-60%) and protein (about 24-36%), and contain various vitamins, calcium, iron, etc. Trace elements are important food and oil crops in many countries in semi-arid and warm regions. The advancement of molecular biotechnology has promoted the development of molecular genetic research of peanut from the initial focus on oil content, quality, and disease resistance to a variety of agronomic traits (Janila, Nigam et al. 2013). Among them, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张晓军颜凡壮于晓娜张绍静司彤邹晓霞王月福王铭伦
Owner QINGDAO AGRI UNIV
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