EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker, kit and identification method for identifying mesona chinensis variety

A technology of molecular markers and kits, applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc., to achieve high reliability, high resolution, and better resolution

Pending Publication Date: 2022-05-17
GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are about 8 to 10 species of Jellygrass found so far, and the above technique is mainly for SSR analysis of 4 species of jelly

Method used

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  • EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker, kit and identification method for identifying mesona chinensis variety
  • EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker, kit and identification method for identifying mesona chinensis variety
  • EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker, kit and identification method for identifying mesona chinensis variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The inventor team designed and synthesized 192 pairs of SSR primers based on the transcriptome data of jelly grass, and screened the identification effect of jelly jelly material. The statistics of some primer sets and identification results are as follows:

[0046]

[0047]

[0048]

[0049]

[0050] Through repeated experiments, it was found that the identification effect of most of the SSR primers designed and synthesized based on transcriptome data was not good. Only the 9 sets of primers shown in the following table are used for better identification of the sites of Xiancao varieties, and the information of the relevant sites and their primer sets is shown in the table below:

[0051]

[0052]Further, the genetic diversity analysis for loci: Na, Ne, I, Ho, He, F, Fst and other parameters were calculated using GenAIex software, and the PIC value of each locus was calculated by Cervus software. The following are the specific analysis results and their ...

Embodiment 2

[0057] The present embodiment provides the identification method of celestial grass kind, comprises the steps:

[0058] S1, provide samples to be tested: BD-1, BD-2, BD-3, FJ-1, FJ-2, FJ-3, YN-1, YN-2, YN-3, TW-1, TW-2 , TW-3, 7-1, 7-2, 7-3, 8-1, 8-2, 8-3, 9-1, 9-2, 9-3, 10-1, 10-2, 10 -3. Please refer to the table below for information on the source of samples of different varieties:

[0059]

[0060]

[0061] S2, extraction of genomic DNA:

[0062] After stacking each material neatly, use scissors to cut an appropriate amount of leaves and place them in a 2mL centrifuge tube. Freeze the centrifuge tubes of the collected samples, use a tissue grinder to fully grind the leaves into powder, and use the Tiangen Plant Genomic DNA Extraction Kit to extract the genomic DNA of the sample to be tested. The concentration of DNA was detected with Nanodrop one, and the DNA was diluted to 10ng / μL with ultrapure water for later use. The sample extraction genomic DNA electrophore...

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Abstract

According to the EST-SSR molecular marker, the kit and the identification method for identifying the mesona chinensis benth variety, EST sequences in transcriptome data are used for analyzing composition and characteristics of microsatellite repetitive sequences contained in the mesona chinensis benth, mesona chinensis benth EST-SSR primers are developed, the effectiveness of the mesona chinensis benth EST-SSR primers and the universality of the mesona chinensis benth EST-SSR primers in mesona chinensis benth are verified, and the mesona chinensis benth variety is identified. The method is expected to provide important tools and valuable information for researches such as genetic relationship identification, phyloevolution, genetic variation and genetic map construction of mesona chinensis and related species thereof.

Description

technical field [0001] The invention relates to the technical field of grass jelly identification, in particular to an EST-SST molecular marker, a kit and an identification method for identifying species of jelly grass. Background technique [0002] Mesona chinensis Benth (Mesona chinensis Benth), also known as jelly grass, jelly grass, is an annual or perennial herb of the Labiatae family, Mesona chinensis Benth, used as a raw material for Chinese herbal medicine and making jelly. Grass jelly has a long history in my country and Southeast Asia. It is distributed in Guangdong, Guangxi, Fujian, Jiangxi, Hainan, Zhejiang, Taiwan and Yunnan. It is also distributed in Vietnam, India, Indonesia and Malaysia (Tang DF, Wei F, Cai ZQ, et al. Analysis of codon usage bias and evolution in the chloroplast genome of Mesona chinensis Benth. DevGenes Evol.). [0003] Grass jelly is mainly wild resources. During the long-term evolution process, grass jelly resources in different regions ha...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125C12Q2563/107Y02A50/30
Inventor 汤丹峰韦范缪剑华全昌乾韦坤华
Owner GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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