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Screening method and application of target gene SSR molecular markers of acanthopanax senticosus

A screening method and molecular marker technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to speed up the breeding process and overcome uncertainties

Active Publication Date: 2018-11-06
DALIAN NATIONALITIES UNIVERSITY
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Problems solved by technology

At present, there is no report on the development of SSR molecular markers for the target gene of Acanthopanax based on RNA-seq technology

Method used

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  • Screening method and application of target gene SSR molecular markers of acanthopanax senticosus
  • Screening method and application of target gene SSR molecular markers of acanthopanax senticosus
  • Screening method and application of target gene SSR molecular markers of acanthopanax senticosus

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Embodiment

[0017] 1. Extract total RNA from Acanthopanax fruit, root, stem and leaf

[0018] One Acanthopanax germplasm SL1 from Suileng County, Heilongjiang Province was used as material. The total RNA of Acanthopanax senticosus from different tissues and organs was extracted according to the method recommended by Shanghai Shenggong Plant RNA Extraction Kit. The total RNA concentration of each tissue and organ was 200-400ng / μL, and stored at -80°C.

[0019] 2. RNA-seq data analysis of Acanthopanax senticosus fruit, root, stem and leaf

[0020] A transcriptome library was constructed by reverse transcription using samples mixed with equal amounts of total RNA from four tissues and organs as templates. Clean reads were obtained by removing adapters, N ratio greater than 0.1% and low-quality reads from the raw reads obtained by sequencing, and the clean reads were spliced ​​using Trinity software to obtain unigene. Compare unigenes with known functional genes in public databases (Nr, Sw...

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Abstract

The invention discloses a screening method and application of target gene SSR molecular markers of acanthopanax senticosus. The screening method specifically includes the following steps that 1, totalRNAs of all kinds of tissue of acanthopanax senticosus are extracted separately; 2, RNA-Seq sequencing is conducted after reverse transcription of the total RNAs is conducted, and Trinity software isused for conducting unigene assembly; 3, unigenes related to synthetic accumulation of acanthopanax senticosus saponins, SOD and the like are screened out, and MicroSatellite software is used for screening SSR sites on the unigenes; 4, Primer3.0 software is used for designing primers, and the primers are synthesized; 5, the extracted genomic DNAs serve as a template for PCR amplification; 6, PCRproducts are subjected to electrophoresis detection, and target gene SSR primers are developed. By means of the method and application, the candidate SSR molecular markers related to target traits areefficiently and more specifically developed, and effective markers are provided for molecular identification of excellent acanthopanax senticosus germplasm resources. An important meaning is achievedfor conducting early identification and speeding up a breeding process of the excellent acanthopanax senticosus germplasm resources.

Description

technical field [0001] The invention belongs to the field of Acanthopanax senticosus molecular marker development and molecular biology technology, and relates to a screening method and application of Acanthopanax senticosus target gene SSR molecular marker. Background technique [0002] Acanthopanax is a deciduous shrub belonging to Araliaceae. The fruit is rich in superoxide dismutase (SOD), which has good free radical scavenging and antioxidant functions; the root bark is rich in saponins, tannins, linolenic acid and various vitamins etc. Both the fruit and the root can be used as medicine, which has the effects of enhancing human immunity, dispelling rheumatism, and strengthening bones and muscles. Acanthopanax is mainly distributed in Heilongjiang, Jilin, Liaoning, Hebei and Shanxi. In recent years, its planting area has gradually expanded. In the cultivation and promotion of Acanthopanax senticosus, the fruit quality of Acanthopanax senticosus is uneven, and the mole...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 丁健阮成江李雪柔韩平杨伞伞
Owner DALIAN NATIONALITIES UNIVERSITY
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