CN108754018A - A kind of screening technique of wilsonii target gene SSR molecular marker and application - Google Patents
A kind of screening technique of wilsonii target gene SSR molecular marker and application Download PDFInfo
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Abstract
The invention discloses a kind of screening technique of wilsonii target gene SSR molecular marker and application, screening technique the specific steps are：1) total serum IgE that extraction wilsonii is respectively organized respectively；2) RNA-seq sequencings are carried out after total serum IgE reverse transcription, Trinity softwares carry out unigene assemblings；3) it filters out with the relevant unigene of the dynamic accumulations such as acanthopanax senticosus saponins, SOD, the sites SSR on these unigene of MicroSatellite software screening methods；4) Primer3.0 software Design primers, and synthesize；5) genomic DNA is that template carries out PCR amplification；6) PCR product carries out electrophoresis detection, develops target gene SSR primers.The present invention is efficient, more targeted exploitation is with the relevant candidate SSR molecular marker of purpose character, and the Molecular Identification for excellent wilsonii germ plasm resource provides significant notation.Excellent wilsonii germ plasm resource early stage is identified and accelerates breeding process and is of great significance.
Description
Technical field
The invention belongs to wilsonii molecular markers development, field of molecular biotechnology, and the present invention relates to a kind of wilsonii mesh
Gene SSR molecular marker screening technique and application.
Background technology
Wilsonii is Araliaceae Acanthopanax machaka, and fruit is rich in superoxide dismutase (SOD), with good clear
Except free radical and anti-oxidation function；Root skin is rich in saponin(e, tannin, leukotrienes and multivitamin etc..Fruit and root can enter
The effect of medicine has and enhances human immunity, wind-damp dispelling, strengthening the bones and muscles.Wilsonii is distributed mainly on Heilungkiang, Jilin, Liaoning, river
North and Shanxi, in recent years, cultivated area is gradually expanded.In wilsonii is cultivated and is promoted, acanthopanax fruit quality is irregular not
Together, it is badly in need of the method for identifying molecules of elite germplasm, scientific basis is provided for high bioactivity ingredient wilsonii Variety Selection.
Microsatellite marker (Simple Sequence Repeats Marker, SSR) is distributed widely in Plant Genome
Different target gene on, repeatability and stability it is preferable.SSR marker has been supplied in plant genetic map construction, diversity point
Analysis and molecular mark etc..In the past, the plant of no reference gene group, in developing SSR molecular labeling frequently with
Est sequence needs to screen and verify from a large amount of primers with the SSR marker of the linkage of characters, heavy workload and with uncertain
Property and blindness.Currently, the report in terms of having no based on RNA-seq technological development wilsonii target gene SSR molecular markers.
Invention content
In order to overcome heavy workload, uncertainty possessed by existing RNA-seq SSR molecular markers screening technique and blind
The deficiency of mesh, the present invention provide a kind of screening technique of wilsonii target gene SSR molecular marker, and the present invention utilizes wilsonii
The RNA-seq data mining target gene SSR molecular markers of fruit, root, stem and leaf are noted using unigene with gene function
The characteristics of releasing and containing the sites SSR, overcomes the shortcomings that prior art screens the uncertainty of SSR marker, blindness, is directed to
Property supplement and wilsonii correlation of attributes candidate SSR molecular marker, and then for wilsonii Germplasm Resources Diversity analysis and property
Shape association analysis provides significant notation, also provides molecule foundation for fine germplasm resources early screening.
The above-mentioned purpose of the present invention is achieved through the following technical solutions：
1. extracting the total serum IgE of acanthopanax fruit, root, stem and leaf, the total serum IgE of mixed in equal amounts different tissues organ respectively；
2. after above-mentioned total serum IgE sample reverse transcription, RNA-seq sequencings are carried out, and carry out using Trinity softwares
Unigene is assembled；
3. emphasis utilizes the annotation of gene function of unigene, screening and acanthopanax senticosus saponins, SOD, vitamin, sugar, acid, fat
The relevant unigene of the dynamic accumulations such as fat acid, and utilize the positions SSR on these unigene of MicroSatellite software screening methods
Point；
4. according to the sites target gene SSR both ends complementary series, using Primer3.0 software Design primers, and synthesize；
5. carrying out PCR amplification by template of wilsonii leaves genomic DNA；
6.PCR products carry out electrophoresis detection, screening meet expected primer size and band clearly target gene SSR draws
Object.
Further, unigene annotations include Nr, Swiss-Prot, KEGG and COG public database in the step 3
Functional annotation.Unigene is mapped to KEGG databases using KAAS functions and obtains metabolic pathway information.According to complicated and number
More unigene annotation informations are measured, related to purpose character and unigene (target gene) containing the sites SSR is filtered out,
And then it is more targeted design and develop with the relevant target gene SSR primers of purpose character, be wilsonii germplasm character
Association analysis provides candidate SSR molecular marker.
Further, the method for the step 6 exploitation target gene SSR marker includes：(1) according to agarose gel electrophoresis
As a result, the availability of preliminary screening SSR primers；(2) polyacrylamide gel electrophoresis is used to detect, analysis target gene SSR draws
The accuracy and applicability of object amplified band.
Compared with the prior art, the invention has the advantages that：The present invention is based on RNA-Seq technological development wilsonii purposes
Gene SSR molecular marker, having organically combined unigene for the first time has the characteristics that annotation of gene function and SSR marker is distributed with, and has
The targetedly relevant SSR primers of dynamic accumulations such as exploitation and acanthopanax senticosus saponins, SOD, vitamin, sugar, acid, aliphatic acid, effectively
Amplification rate reaches 52.3%.The target gene SSR marker obtained can be used as analysis of genetic diversity, fingerprint map construction and property
The candidate functional label of shape association analysis, the very rare wilsonii SSR marker quantity of supplement are excellent wilsonii germ plasm resource
Molecular Identification provide significant notation.The shortcomings that overcoming uncertainty, the blindness of prior art screening SSR marker, to excellent
Good wilsonii germ plasm resource early stage identifies and accelerates that breeding process is of great significance.
Specific implementation mode
The present invention is described in detail below by specific embodiment, but is not limited the scope of the invention.Unless otherwise specified, originally
Experimental method is conventional method used by invention, and experiment equipment used, material, reagent etc. commercially obtain.
Embodiment
1. extracting acanthopanax fruit, root, stem and leaf total serum IgE
Using 1 wilsonii germplasm SL1 of Heilongjiang Province Suiling County as material.Work plant RNA extraction reagent is given birth to according to Shanghai
Box recommends the total serum IgE of method extraction wilsonii different tissues organ.The total rna concentration 200-400ng/ μ L of each histoorgan, -80
DEG C preserve.
2. the RNA-seq data analyses of acanthopanax fruit, root, stem and leaf
Reverse transcription, structure transcript profile library are carried out by template of the sample of 4 histoorgan total serum IgE mixed in equal amounts.It will survey
The raw reads that sequence obtains remove connector, N ratios obtain clean reads after being more than 0.1% and low-quality reads, utilize
Trinity softwares are spliced to obtain unigene to clean reads.By unigene and public database (Nr, Swiss-
Prot, KEGG and COG etc.) in known function gene compared.Unigene is mapped to KEGG data using KAAS functions
Library obtains metabolic pathway information.
3. the relevant unigene of the synthesis such as screening and saponin(e, SOD, vitamin, sugar, acid, aliphatic acid (and contain SSR
Point)
According to unigene annotations, metabolic pathway information and pertinent literature report, screening is given birth to acanthopanax senticosus saponins, SOD, dimension
Element, sugar, acid, aliphatic acid etc. synthesize relevant unigene.The target gene screened includes and the relevant SS of saponin formation
(squalene synthase)、HMGR(3-hydroxy-3-methylglutaryl coenzyme A)、SE(squalene
Epoxidase), DXPS (1-deoxy-D-xylulose-5-phosphate synthase) and OSCs (epoxy squalene
Cyclase) base；Synthesized with lipid relevant acetyl-coA carboxylase, 3-ketoacyl-ACP-synthase II,
3-ketoacyl-ACP-synthase III、fatty acyl-ACP thioesterase B、fatty acyl-ACP
thioesterase A、stearoyl-CoA desaturase、oleate delta-12desaturase、glycerol-3-
phosphate acyltransferase、diacylglycerol acyltransferase、mitochondrial trans-
2-enoyl-CoA reductase, 3-ketoacyl-CoA synthase and phosphatidic acid phosphatase
Gene；Relevant fructose-bisphosphate aldolase, citrate synthase, malate are synthesized with saccharic acid
dehydrogenase、phosphoenolpyruvate carboxylase、alpha-amylase、glucosidase、
Sucrose synthase, neutral invertase, pectinesterase, polygalacturonase gene；With dimension
Raw element synthesizes related TC (tocopherol cyclase), GDP-mannose pyrophosphorylase and Myo-
Inositol oxygenase genes；And sod gene.Using the SSR of these unigene of MicroSatellite software detections
Site.
4. carrying out PCR amplification by template of wilsonii leaves genomic DNA
Wilsonii leaves genomic DNA is extracted according to improved Tiangeng company plant genome DNA extracts reagent cassette method,
Specific method is：1. blade (20~40mg) moves into centrifuge tube after fully milling, 400~600 μ L buffer solutions LP1 and 4~6 are added
Vortex oscillation 1min after μ L RNase A；2. vortex oscillation 1min after 100~200 μ L buffer solutions LP2 is added；3. 12000rpm from
Heart 5min shifts supernatant；4. oscillation mixing 15s after the buffer solution LP3 of 1.5 times of volumes is added；It is inhaled 5. all solution are moved into
The centers attached column CB3,12000rpm centrifuges 2min after placing 10min, abandons waste liquid；6. 600 μ L rinsing liquids are added in adsorption column CB3
PW, 13000rpm centrifuge 1.5min, abandon waste liquid；7. repeating step 6.；8. adsorption column CB3 puts back to collecting pipe, 12000rpm centrifugations
After 2min, CB3 is placed at room temperature for 15min；9. adsorption column CB3 moves into new centrifuge tube, 36 are vacantly added dropwise to adsorbed film central part
~50 μ L TE solution, are placed at room temperature for 5min, and 12000rpm centrifuges 2min；DNA solution is taken to be further dropped into adsorbed film center, room temperature
2min is placed, 12000rpm centrifuges 2min, obtains acanthopanax piece Genomic DNA solution.
According to the objective gene sequence containing the sites SSR in step 3,107 pairs of primers are devised, with acanthopanax piece gene
Group DNA is that template carries out PCR amplification, and reaction total system is 20 μ L：The dNTP 0.4 of 10 × PCR buffer, 2 μ L, 10mmol/L
μ L, each 1 μ L of upstream and downstream primer of 10 μm of ol/L, 2 μ L of template DNA of 20ng/ μ L, Taq enzyme 0.2 μ L, ddH2O 13.4μL；Instead
The program is answered to be：94 DEG C of pre-degeneration 4min；94 DEG C of denaturation 30s, 58 DEG C of annealing 30s, 72 DEG C of extension 30s, 35 cycles；72 DEG C of extensions
10min；4 DEG C of preservations.
5. electrophoresis detection PCR product develops target gene SSR molecular marker
The PCR product of 107 pairs of SSR primers is detected first with 2.5% agarose gel electrophoresis, preliminary screening obtains 56 pairs
SSR marker suitable for wilsonii.Then, this 56 PCR further are detected using 8% polyacrylamide gel vertical slab electrophoresis
Amplified production (table 1) also obtains expected primer size and clearly band.
The wilsonii target gene SSR primers that table 1 is developed based on RNA-Seq
Claims (9)
1. a kind of screening technique of wilsonii target gene SSR molecular marker, which is characterized in that screening technique includes following step
Suddenly：
Step 1, the total serum IgE for extracting acanthopanax fruit, root, stem and leaf respectively, the total serum IgE of mixed in equal amounts different tissues organ；
Step 2 after the total serum IgE sample reverse transcription in step 1, will be carried out RNA-seq sequencings, and be carried out using Trinity softwares
Unigene is assembled；
Step 3 has the characteristics that annotation of gene function using unigene, filters out and wilsonii functional active components composition product
Tire out relevant unigene, utilizes the sites SSR on MicroSatellite software screening methods these unigene；
Step 4, according to the sites target gene SSR both ends complementary series, using Primer3.0 software Design primers, and synthesize；
Step 5 carries out PCR amplification by template of wilsonii leaves genomic DNA；
Step 6, PCR product carry out electrophoresis detection, screening meet expected primer size and band clearly target gene SSR draws
Object.
2. screening technique according to claim 1, which is characterized in that in the step 1, each histoorgan of wilsonii
Total rna concentration 200-400ng/ μ L, -80 DEG C of preservations.
3. screening technique according to claim 1, which is characterized in that in the step 2, obtained raw will be sequenced
Reads removes connector, N ratios obtain clean reads after being more than 0.1% and low-quality reads, utilizes Trinity softwares
Clean reads are spliced to obtain unigene, the known function gene in unigene and public database is carried out pair
Than unigene is mapped to KEGG databases using KAAS functions and obtains metabolic pathway information.
4. screening technique according to claim 1, which is characterized in that more according to complicated and quantity in the step 3
Unigene annotation informations, filter out and the wilsonii relevant unigene of functional active components dynamic accumulation, use
The sites SSR of these wilsoniis of MicroSatellite software detections unigene, so it is more targeted design and develop with
The relevant target gene SSR primers of purpose character provide candidate SSR molecular marker for the trait associations analysis of wilsonii germplasm.
5. screening technique according to claim 1, which is characterized in that in the step 5, extract acanthopanax piece gene
Group DNA, specific method are：1. blade weighs and moves into centrifuge tube after 20~40mg fully mills, 400~600 μ L buffer solutions are added
Vortex oscillation 1min after LP1 and 4~6 μ L RNase A；2. vortex oscillation 1min after 100~200 μ L buffer solutions LP2 is added；③
12000rpm centrifuges 5min, shifts supernatant；4. oscillation mixing 15s after the buffer solution LP3 of 1.5 times of volumes is added；5. will own
Solution moves into the centers adsorption column CB3, and 12000rpm centrifuges 2min after placing 10min, abandons waste liquid；6. being added 600 in adsorption column CB3
μ L rinsing liquids PW, 13000rpm centrifugation 1.5min, abandons waste liquid；7. repeating step 6.；8. adsorption column CB3 puts back to collecting pipe,
After 12000rpm centrifuges 2min, CB3 is placed at room temperature for 15min；9. adsorption column CB3 moves into new centrifuge tube, to adsorbed film central portion
Position is hanging to be added dropwise 36~50 μ L TE solution, is placed at room temperature for 5min, and 12000rpm centrifuges 2min；DNA solution is taken to be further dropped into absorption
Film center is placed at room temperature for 2min, and 12000rpm centrifuges 2min, obtains acanthopanax piece Genomic DNA solution.
6. screening technique according to claim 1, which is characterized in that in the step 5, be with leaves genomic DNA
Template carries out PCR amplification, and reaction total system is 20 μ L：0.4 μ L of dNTP, 10 μ of 10 × PCR buffer, 2 μ L, 10mmol/L
Each 1 μ L of upstream and downstream primer of mol/L, 2 μ L of template DNA of 20ng/ μ L, Taq enzyme 0.2 μ L, ddH2O 13.4μL；
Response procedures are：94 DEG C of pre-degeneration 4min；94 DEG C of denaturation 30s, 58 DEG C of annealing 30s, 72 DEG C of extension 30s, 35 cycles；72
DEG C extend 10min；4 DEG C of preservations.
7. screening technique according to claim 1, which is characterized in that in the step 6, exploitation target gene SSR marks
The method of note includes：(1) using agarose gel electrophoresis as a result, the availability of preliminary screening SSR primers；(2) polypropylene is used
Acrylamide gel electrophoresis detection analyzes the accuracy and applicability of target gene SSR primer amplification bands.
8. the wilsonii target gene SSR molecular marker that the screening technique according to claim 1-7 obtains is in wilsonii
Germplasm Resources Diversity is analyzed and trait associations analysis, the application of fine germplasm resources early screening.
9. the wilsonii target gene SSR primers of screening technique exploitation according to claim 1 are：
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109486999A (en) * | 2018-12-25 | 2019-03-19 | 云南农业大学 | The application of Radix Notoginseng SSR polymorphism primer, pleiomorphism detecting method and SSR marker on total saponin content determines |
| CN109486997A (en) * | 2018-12-25 | 2019-03-19 | 云南农业大学 | Purposes of the Radix Notoginseng SSR marker on ginsenoside Rb1's content determines |
| CN114214429A (en) * | 2021-12-18 | 2022-03-22 | 山西农业大学 | Development method of microsatellite markers of icerya purchasi |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120039328A (en) * | 2010-10-15 | 2012-04-25 | 대한민국(농촌진흥청장) | Ssr primer and kits derived from acanthopanax senticosus, and use of there |
| CN103642912A (en) * | 2013-11-29 | 2014-03-19 | 中国农业科学院作物科学研究所 | Method for developing mung bean simple sequence repeat (SSR) primer based on transcriptome sequencing |
| CN103923909A (en) * | 2013-01-15 | 2014-07-16 | 复旦大学 | Specific molecular marker for panax ginseng and panax quinquefolius and identification method thereof |
| CN107201408A (en) * | 2017-07-15 | 2017-09-26 | 中国热带农业科学院南亚热带作物研究所 | A kind of method that exploitation sisal hemp SSR primers are sequenced based on transcript profile |
| CN108034696A (en) * | 2018-02-02 | 2018-05-15 | 中南大学 | A kind of method based on transcript profile sequencing SSR primers development |
| CN108192893A (en) * | 2017-08-31 | 2018-06-22 | 中国热带农业科学院热带作物品种资源研究所 | The method of exploitation Blumea balsamifera SSR primers is sequenced based on transcript profile |
-
2018
- 2018-07-27 CN CN201810852453.5A patent/CN108754018B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120039328A (en) * | 2010-10-15 | 2012-04-25 | 대한민국(농촌진흥청장) | Ssr primer and kits derived from acanthopanax senticosus, and use of there |
| CN103923909A (en) * | 2013-01-15 | 2014-07-16 | 复旦大学 | Specific molecular marker for panax ginseng and panax quinquefolius and identification method thereof |
| CN103642912A (en) * | 2013-11-29 | 2014-03-19 | 中国农业科学院作物科学研究所 | Method for developing mung bean simple sequence repeat (SSR) primer based on transcriptome sequencing |
| CN107201408A (en) * | 2017-07-15 | 2017-09-26 | 中国热带农业科学院南亚热带作物研究所 | A kind of method that exploitation sisal hemp SSR primers are sequenced based on transcript profile |
| CN108192893A (en) * | 2017-08-31 | 2018-06-22 | 中国热带农业科学院热带作物品种资源研究所 | The method of exploitation Blumea balsamifera SSR primers is sequenced based on transcript profile |
| CN108034696A (en) * | 2018-02-02 | 2018-05-15 | 中南大学 | A kind of method based on transcript profile sequencing SSR primers development |
Non-Patent Citations (9)
| Title |
|---|
| KYUNG JUN LEE ET AL: "Development of Microsatellite Markers and their Use in Genetic Diversity and Population Analysis in Eleutherococcus senticosus", 《KOREAN J. PLANT RES.》 * |
| 冯延芝等: "基于ＲＮＡ－Ｓｅｑ的杜仲转录组微卫星特征分析", 《中国农业大学学报》 * |
| 刘灵智等: "柑橘果实成熟过程中糖酸代谢及其关键酶的研究进展", 《园艺学文集》 * |
| 匿名: "第29章脂类的生物合成", 《道客巴巴》 * |
| 尚增振等: "高等植物中维生素C合成及其相关酶研究进展", 《中国科技论文在线》 * |
| 李雪柔等: "RNA-seq 技术开发刺五加目的基因SSR标记", 《分子植物育种》 * |
| 潘卫东等: "植物维生素E 合成相关酶基因的克隆及其在体内功能研究进展", 《植物学通报》 * |
| 许晓双等: "三萜皂苷生物合成途径及关键酶的研究进展", 《世界科学技术中医药现代化》 * |
| 闫蕊等: "RNA-seq 技术开发油茶目的基因SSR 标记", 《分子植物育种》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109486999A (en) * | 2018-12-25 | 2019-03-19 | 云南农业大学 | The application of Radix Notoginseng SSR polymorphism primer, pleiomorphism detecting method and SSR marker on total saponin content determines |
| CN109486997A (en) * | 2018-12-25 | 2019-03-19 | 云南农业大学 | Purposes of the Radix Notoginseng SSR marker on ginsenoside Rb1's content determines |
| CN109486997B (en) * | 2018-12-25 | 2022-04-19 | 云南农业大学 | Application of pseudo-ginseng SSR marker in determination of ginsenoside Rb1 content |
| CN114214429A (en) * | 2021-12-18 | 2022-03-22 | 山西农业大学 | Development method of microsatellite markers of icerya purchasi |
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| CN108754018B (en) | 2022-03-04 |
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