Kit and method for identifying hybrid progeny of citrus and pomelo plants using InDel marker

A kit, citrus technology, applied in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., can solve problems such as stability and repeatability defects, reduced identification accuracy, and cumbersome operation process.

Active Publication Date: 2019-01-25
HUAZHONG AGRI UNIV
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AI Technical Summary

Problems solved by technology

Randomly amplified polymorphic DNA (RAPD) is a molecular technique based on PCR for polymorphism analysis of the genome, and its amplification stability and repeatability are seriously defective; amplified fragment length polymorphism marker technology (AFLP ) is often used to identify germline DNA fingerprints, genetic diversity of fruit tree varieties, etc., but it is prone to false positive fragments, which reduces the accuracy of identification; simple repeat sequence (SSR) is a molecular marker technology that is relatively widely used at present, and its markers are distributed throughout the world. There are high polymorphisms in the genome, but the marker bands need to be distinguished by polyacrylamide gel electrophoresis. The operation process is cumbersome and involves some toxic reagents, so it is not suitable for safe and efficient identification.
[0006] The above-mentioned technical methods have some defects and limit their application in hybrid identification work. Therefore, at this stage, an accurate, economical and efficient hybrid identification method is needed in order to improve the efficiency of scientific research or breeding process

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  • Kit and method for identifying hybrid progeny of citrus and pomelo plants using InDel marker
  • Kit and method for identifying hybrid progeny of citrus and pomelo plants using InDel marker
  • Kit and method for identifying hybrid progeny of citrus and pomelo plants using InDel marker

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Embodiment Construction

[0022] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0023] 1. InDel primer design and screening

[0024] 1.1 Material selection

[0025] Nine pomelo varieties were selected as hybrid parents, as shown in Table 1, and then the nine pomelo varieties were combined in pairs to obtain offspring. The genomic DNA of the female parent and the progeny of the respective hybrid combinations were extracted and purified by the CTAB micromethod to meet the quality requirements for resequencing.

[0026] Table 1 Information of 9 pomelo varieties

[0027]

[0028] 1.2 Design of InDel labeled primer pairs and screening of effective primer pairs

[0029] Based on the genome resequencing data of the above nine pomelo varieties, specific high-quality primer pairs were designed, an...

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Abstract

The present invention relates to a kit for identifying hybrid progeny of citrus and pomelo plants using InDel markers, comprising one or more primer pairs of 17 primer pairs; at method for identify hybrid progeny of citrus and pomelo plants use InDel markers, includes the following steps: the genomes of the progenies of citrus and pomelo plants to be identified were obtained, as well as the expected male and female genomes, primer pairs were used to amplify the corresponding InDel markers in the genomes of the progeny of the citrus and pomelo plants to be identified and the expected male and female genomes, the PCR amplification products were electrophoresed in agarose gel and the hybrids were identified according to the electrophoretic banding patterns. The kit and the method of the invention can be easily and quickly applied to scientific research or cross breeding practice of related pomelo varieties, and provide effective methods and ideas for breeding work of other pomelo varieties and lay a good foundation.

Description

technical field [0001] The invention relates to the fields of citrus planting and breeding, and more particularly relates to a kit and an identification method for identifying hybrid progenies of citrus pomelo plants using InDel markers. Background technique [0002] Citrus is the largest category of fruit in the world, and its industrial economic value is huge, among which Citrus grandis is a very important part. The planting area and output of pomelo in my country ranks first in the world. Shatian pomelo, Guanxi honey pomelo, Pingshan honey pomelo, Dianjiang white pomelo and other Chinese traditional fine pomelo varieties are the main ones, and the planting area is relatively large. With the rapid development of pomelo industry, there are some problems in scientific research and industry that need to be solved urgently. my country's pomelo varieties and technical support are still in the initial stage of development, with many varieties and unreasonable production structu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 柴利军曹宗洪梁梅邓秀新
Owner HUAZHONG AGRI UNIV
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