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A method and indel molecular marker for traceable origin of maize mixed pollination hybrids

A technology of molecular markers and mixed pollination, which is applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of manpower consumption, process management difficulties, and influence on breeding process, and achieve stable detection results, low detection cost, The effect of detecting convenience

Active Publication Date: 2022-06-07
JIANGSU ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This process requires thousands or even tens of thousands of crosses during the pollination period of the male parent due to the method of single-plant hybridization, which consumes a lot of manpower.
Due to the large number of inbred lines and hybrid combinations, the process management is difficult, often resulting in some target combinations not being successfully assembled, which affects the entire breeding process

Method used

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  • A method and indel molecular marker for traceable origin of maize mixed pollination hybrids
  • A method and indel molecular marker for traceable origin of maize mixed pollination hybrids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Development and Validation of an InDel Molecular Marker Traceable to the Origin of Maize Mixed-pollinated Hybrids:

[0026] Download the B73 reference genome sequence (V4) from the maizeGDB (https: / / www.maizegdb.org / ) website, and download 205 maize autogenous sequences from the NCBI (https: / / www.ncbi.nlm.nih.gov / ) website Inbred genome resequencing data (PRJNA82843, SRP011907 and PRJNA260788). The genome resequencing data were subjected to Q20 (Phred-Score≥20, ie 1% error rate) quality control and sequencing length filtering (L20, length≥20bp) by SoleaQ software package. The cleaned up sequencing data were assembled de novo by SOAPdenovo2 software.

[0027] The mInDel software was used to identify the InDel variation between the assembled sequence and the B73 reference genome and to design primer sequences. Primers were selected according to the conditions of unique InDel genome location, large InDel differences, and high primer design scores, and the co-dominant mol...

Embodiment 2

[0030] A method for traceability of the source of mixed-pollinated hybrids of corn, comprising the steps of:

[0031] (1) Combination of hybrids to be tested:

[0032] Twenty-five parental inbred lines were planted in the experimental field, and the tassel was removed from the female parent row before tasseling. The pollination method is natural open pollination in the field. The ear after pollination is listed for harvest, and after threshing, it is planted as a single seed, and the parent is planted at the same time. Seeds from the same ear were planted together for later identification of hybrid male parents by phenotypic comparison.

[0033] (2) Obtaining the genotypes of the parents and the hybrid to be tested:

[0034] Using the 37 high-quality InDel molecular markers verified in Example 1, 25 parental inbred lines and 49 randomly selected hybrids to be tested were genotyped by agarose gel electrophoresis. The short band of each molecular marker was recorded as A, th...

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Abstract

The invention relates to the technical field of molecular markers and molecular breeding, in particular to a method for tracing the source of corn mixed pollinated hybrids and InDel molecular markers, comprising the following steps: 1) extracting all male parents, female parents and mixed pollinated hybrids to be tested Leaf DNA; 2) Use the DNA of the male parent and the female parent to screen the molecular markers that can establish fingerprints; 3) Use the screened molecular markers to perform PCR amplification on the DNA template in step 1), and record the male parent, the female parent and the DNA template to be tested. The genotype of mixed pollination hybrids; 4) deduce all possible hybrid genotypes according to the genotypes of the male parent and female parent obtained; 5) step 3) obtains the mixed pollination hybrid genotypes to be checked and step 4) derivation compare the genotypes of all the hybrids, and record the number of loci of the same genotype; It is the parental source of the mixed pollination hybrid to be tested.

Description

technical field [0001] The invention relates to the technical field of molecular markers and molecular breeding, in particular to a method for tracing the source of hybrid corn mixed pollination. Background technique [0002] Corn is the largest grain and feed crop in my country, which plays an important role in ensuring food security and improving people's quality of life. With the reduction of the total amount of arable land in my country, the sown area of ​​corn has also decreased. Therefore, the improvement of corn yield is the key to ensure the total corn yield. In addition to optimizing cultivation measures, the upgrading of corn varieties is also an effective way to increase the yield per unit of corn. my country invests a lot of money every year for the cultivation of new varieties of corn. [0003] At present, the cultivation of new maize varieties relies on a large number of combinations of inbred lines and multi-point combination identification for many years. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张体付钱益亮赵涵阮龙梁帅强林峰陆海燕
Owner JIANGSU ACAD OF AGRI SCI
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