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Specific DNA molecular marker for populus euphratica sex determination developed based on BSA mixed pool sequencing analysis

A molecular marker, Populus euphratica technology, applied in the field of bioinformatics, can solve the problems of development and application of molecular markers for sex identification of Populus euphratica that have not yet been seen

Pending Publication Date: 2021-03-02
TARIM UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the plants of the genus Populus, currently only the invention patent with the authorized announcement number CN201911012229.6 discloses two female-specific DNA molecular markers for early sex identification of Populus alba, but there is no development and research on molecular markers related to sex identification of Populus euphratica. application

Method used

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  • Specific DNA molecular marker for populus euphratica sex determination developed based on BSA mixed pool sequencing analysis
  • Specific DNA molecular marker for populus euphratica sex determination developed based on BSA mixed pool sequencing analysis
  • Specific DNA molecular marker for populus euphratica sex determination developed based on BSA mixed pool sequencing analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Assembly of Populus euphratica male and female genomes

[0024] High-quality genomic DNA was extracted from leaves of Populus euphratica, and libraries were constructed for Illumina, PacBio and Hi-C sequencing. Preliminary assembly was performed based on the reads obtained by PacBio sequencing, and the assembly results were corrected using the reads obtained by Illumina sequencing to obtain a draft version of the genome. Use Hi-C sequencing technology to assemble the male and female genomes, annotate and evaluate the male and female genomes, and obtain high-quality male and female genome sequences at the chromosome level.

Embodiment 2

[0025] Example 2 Development of sex-related candidate DNA molecular markers in Populus euphratica:

[0026] Using the male genome as a reference, association analysis was performed on the BSA-seq data of 96 female plants and 97 male plants, and the region where the female mixed pool depth was continuously 0 and the male mixed pool depth was continuously greater than 10 was obtained. Sequences including upstream and downstream 150bp were extracted from these regions as candidate gender markers. Blast was performed on the obtained markers and the assembled female plant genome, and the 30 candidate markers obtained were preliminarily classified into 16 markers shared by both sexes, 10 markers that may be shared by both sexes, and 4 markers unique to males.

Embodiment 3

[0027] Example 3 Candidate Sex Marker Primer Design and Marker Screening

[0028] Using Primer 5 software, 30 candidate sex markers were used to design primers for the target fragments. The length of the primers was 18-22bp, and the GC% was 35-48%. Using 30 pairs of primers and using the genomic DNA of Populus euphratica as a template, the corresponding markers were verified and screened by PCR. Finally, 3 preferred shared markers F1-F3 were screened from 16 markers shared by both sexes. A total of 14 markers were screened to obtain 4 male-specific markers M1-M4, as shown in Table 1.

[0029] Table 1 Sex markers and primer sequences of Populus euphratica

[0030]

[0031] Example 4 Sex Identification of Populus euphratica

[0032] 1. Experimental materials

[0033] There were 10 male and female Populus euphratica whose genders were known from different locations, as shown in Table 2.

[0034] Table 2 Populus euphratica samples from 10 locations

[0035]

[0036] ...

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Abstract

The invention develops the specific DNA molecular marker for populus euphratica sex identification on the basis of BSA mixed pool sequencing analysis, and designs corresponding detection primers on the basis of sequence information of the specific DNA molecular marker. The sex of populus euphratica from different places can be distinguished economically, rapidly, accurately and sensitively, technical support is provided for work of selectively transplanting female plants or male plants of populus euphratica in nursery stock breeding, and theoretical support is also provided for popularizationwork of desertification control of populus euphratica.

Description

technical field [0001] The invention relates to the field of biological information technology and the field of biotechnology, in particular to the sequencing analysis of Populus euphratica BSA mixed pool and the development of specific DNA molecular markers for sex identification of Populus euphratica. Background technique [0002] Populus euphratica Oliv. is an important arbor tree species in desert areas. It is mainly distributed in Northwest China, and Xinjiang is the most widely distributed. It has strong drought resistance, cold resistance, and salt-alkali resistance. It plays an irreplaceable role in many aspects such as balance, containment of desert expansion, protection of biodiversity, and protection of ecological safety of industrial and agricultural production. In Xinjiang, Populus euphratica is often the first choice for urban landscaping and shelterbelt construction. In recent years, due to the influence of natural and human factors such as land reclamation, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6879C12Q1/6809C12N15/11
CPCC12Q1/6895C12Q1/6879C12Q1/6809C12Q2600/13C12Q2600/166C12Q2535/122C12Q2537/165Y02A40/81
Inventor 李志军吴智华张山河刘虹曲文蕊覃瑞翟军团韩晓莉
Owner TARIM UNIV
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