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Molecular marker primers for identifying liquidambar plants, application of molecular marker primers and identification method of liquidambar plants

A technology assisted by molecular markers and molecular markers, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve problems such as constraints and difficult species judgments

Active Publication Date: 2020-06-26
BEIJING FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0007] For the identification of liquidambar species, for a long time, biologists have mainly divided different species through morphological and anatomical evidence, but these phenotypic characteristics are easily affected by environmental factors and developmental stages, coupled with the general convergence effect caused by natural selection And parallel evolution factors, etc., make the judgment of species difficult. If you cannot efficiently and accurately identify different liquidambar species and confirm the relationship between them, there will be a certain degree of blindness in the parental selection of hybrid breeding. For the creation of this economic tree species New Breed Existence Bondage

Method used

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  • Molecular marker primers for identifying liquidambar plants, application of molecular marker primers and identification method of liquidambar plants
  • Molecular marker primers for identifying liquidambar plants, application of molecular marker primers and identification method of liquidambar plants
  • Molecular marker primers for identifying liquidambar plants, application of molecular marker primers and identification method of liquidambar plants

Examples

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Embodiment 1

[0135]Embodiment 1, design and development of Liquidambar genus-specific SSR primers

[0136] The plant material used in this example is a sample of sweetgum sweetgum, which was taken from the excellent tree population of sweetgum sweetgum (53 samples) in Jigong Mountain, Xinyang, Henan Province in early April 2017, and DNA was extracted from young leaves of sweetgum sweetgum that were robust and free from diseases and insect pests .

[0137] 1. Design of SSR primers

[0138] Liquidambar transcriptome sequence data (accession numbers: SRR1514949, SRR1514913) were downloaded from the SRA (Sequence Read Archive) database of NCBI (National Center of Biotechnology Information). Entrusted Guangzhou Jidio Biotechnology Co., Ltd. to assemble the downloaded sequence reads, and then searched the gene SSR sites from the Unigene library through the MISA program, and the search criteria were: two, three, four, five, and six nucleotides The minimum number of repeats was 9, 6, 5, 5, and 4...

Embodiment 2

[0170] Embodiment 2, the application of specific SSR primer pair (verification of Liquidambar molecular marker)

[0171] The plant materials used in this example are from April 2017. The branches of 53 Chinese sweetgum trees were selected from Jigongshan, Xinyang, Henan, and the young leaves were collected and stored in a -80°C refrigerator for later use. The DNA was extracted by the CTAB method ( References: Doyle, J.J.; Doyle, J.L.A rapid DNA isolation procedure for small quantities of freshleaf tissue. Phytochem Bull. 1987, 19, 11–15.), and finally obtained 53 samples of Chinese sweetgum.

[0172] In addition, in order to test the universality of the primers used among species, 6 sweetgum sweetgum plants (collected from Shanghai Chenshan Botanical Garden) and 13 sweetgum sweetgum hybrids cultivated by Beijing Forestry University (male parent is Chinese sweetgum sweetgum, female parent Universal detection of primers for Liquidambar nigrum. The young leaves of sweetgum sweet...

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Abstract

The invention discloses molecular marker primers for identifying liquidambar plants, an application of the molecular marker primers and an identification method of the liquidambar plants. In the invention, a transcriptome sequence obtained by high-throughput sequencing is used as a basis for development of a liquidambar EST-SSR marker, transcriptome sequence reads are downloaded from a National Center for Biotechnology Information (NCBI) database and are assembled to form Unigenes; MISA software is used to find SSR sites, and a Primer3 is used to design primers; polymorphic SSR primers are screened to obtain polymorphic primer information; and finally, 32 pairs of polymorphic SSR primers are obtained. The primers provided by the invention can assist in the identification of liquidambar variety, can be used as a powerful research tool in germplasm resource evaluation, genetic map construction, molecular-assisted selection breeding, genetic diversity analysis and the like, and lays a foundation for further studies on the genetics and breeding of the liquidambar plants.

Description

technical field [0001] The present invention relates to a molecular marker primer for identification of plants, its application and plant identification method, in particular to an EST-SSR molecular marker primer for identification of sweetgum plant, its application and plant identification method. The present invention belongs to the molecular biology of plant varieties The technical field of scientific identification methods. Background technique [0002] The species of Liquidambar spp. grow fast, have good material properties and are suitable for mountain afforestation, especially the North American sweetgum (L. styraciflua) and sweetgum (L. formosana). In addition, as my country's national economic level improves day by day, the demand for tree species with higher ornamental value is also increasing day by day. Sweetgum and sweetgum are famous ornamental tree species with high development value. Liquidambar tree species are deciduous trees of the suborder Hamamelidacea...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156Y02P60/40Y02A40/22
Inventor 张金凤陈思源董明亮赵健范英明孔立生
Owner BEIJING FORESTRY UNIVERSITY
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