CN106011134A - Co-separation molecular marker TCR540 of clubroot resistant CRb gene of Chinese cabbages, primers and application - Google Patents
Co-separation molecular marker TCR540 of clubroot resistant CRb gene of Chinese cabbages, primers and application Download PDFInfo
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Abstract
The invention discloses a co-separation molecular marker TCR540 of a clubroot resistant CRb gene of Chinese cabbages. A nucleotide sequence of the co-separation molecular marker TCR540 is represented as SEQ ID NO.1, TCR540 and the CRb gene are co-separated, and TCR540 can be applied to molecular marker assisted selection of the clubroot resistant CRb genes of Brassica including Chinese heading cabbages and green pakchoi. The markers are used simultaneously in an assisted selection process, the theoretical accuracy of the selection can be up to 100%, and the co-separation molecular marker TCR540 is good in repeatability, high in reliability and low in detection cost and saves time and labor.
Description
Technical field
The invention belongs to biological technical field, be specifically related to being divided into from molecule of a kind of Chinese cabbage anti-clubroot CRb gene
Labelling TCR540, primer and application.
Background technology
2002, this seminar obtained Chinese cabbage Doubled haploid line ' the CR Shinki DH ' of 1 part of anti-Caulis et Folium Brassicae campestris clubroot
System, research shows this disease-resistant material anti-Caulis et Folium Brassicae campestris plasmodiophora brassicae biological strain 2,4 and 8 (Piao etc., Conversion of AFLP
Marker linked to clubroot resistance gene into SCAR marker.2002, J Kor Soc
Hort Sci 43:653–665).Research proves that anti-clubroot gene is dominant single-gene further, and named CRb (Piao
Deng, SCAR and CAPS mapping of CRb, a gene conferring resistance to
Plasmodiophora brassicae in Chinese cabbage (Brassica rapa ssp.pekinensis),
Theor Appl Genet, 2004,108:1458-1465).
During Vegetable Crops of Brassica anti-clubroot kind transformation, per in generation, is required for using the method mirror of Field inoculation pathogen plasmodiophora
Determine the genotype of intermediate materials, and environmental condition influences whether the selection of correct intermediate materials, so certainly will affect transformation and enter
Journey, the problem that therefore disease-resistant variety transformation is difficult the most thoroughly solves.Molecular Marker Assisted Selection Technology (MAS) is probably finally
Solve the effective way of this problem.
At present, domestic Agricultural University Of Shenyang Piao Zhong cloud seminar to obtain clubroot gene C Rb anti-with Vegetable Crops of Brassica chain
Labelling (2004,2014), including TCR01, TCR05 and TCR09, is shown in (Piao etc., SCAR and CAPS mapping of
CRb,a gene conferring resistance to Plasmodiophora brassicae in Chinese
Cabbage (Brassica rapa ssp.pekinensis), and TCR108, TCR30, TCR74, TCR79, be shown in (Zhang
Deng, Fine genetic and physical mapping of the CRb gene conferring resistance to
Clubroot disease in Brassica rapa., Mol Breeding, 34:1173-1183).But these labellings are not
Be with CRb be divided into from labelling, utilize these labellings to carry out during molecular marking supplementary breeding and easily cause Linkage drag.
Summary of the invention
It is an object of the invention to provide being divided into from molecular marker TCR540, drawing of a kind of Chinese cabbage anti-clubroot CRb gene
Thing and application, utilize this molecular marker that plant to be measured can carry out seedling stage assay, thus reduce Linkage drag, simplifies Chinese cabbage and resists
The transformation program of clubroot kind.
The invention provides being divided into from molecular marker TCR540, described DABAI of a kind of Chinese cabbage anti-clubroot CRb gene
Dish anti-clubroot CRb gene be divided into the nucleotide sequence from molecular marker TCR540 as shown in SEQ ID NO.1, for:
5’-TGACATTGTTGATGTGCTGATAAAAAAATCAGTAAGTCTAAGCAAATGTATATATTTTAGTTCTAT
TGGAACTTTGGATTACTGCTTTCTTCGCTTAGAAAAGTTTGCATTTTTACTTCAGGGAACCGAAGCAATTGAAGGCA
TATTT-3’。
Present invention also offers being divided into from molecular marker TCR540's of a kind of above-mentioned Chinese cabbage anti-clubroot CRb gene
PCR specificity amplification primer, including:
F TCR540:5’-TGACATTGTTGATGTGCTGA-3’
R TCR540:5’-AAATATGCCTTCAATTGCTTC-3’。
Present invention also offers a kind of Chinese cabbage anti-clubroot CRb gene is divided into from molecular marker TCR540, at molecule
Application in marker assisted selection.
Present invention also offers a kind of Chinese cabbage anti-clubroot CRb gene is divided into from molecular marker TCR540, in DABAI
Application in dish anti-clubroot molecular marker assisted selection.
Present invention also offers a kind of above-mentioned Chinese cabbage anti-clubroot CRb gene is divided into answering from molecular marker TCR540
By method, specifically include:
(1) genomic DNA of detected materials is extracted by CTAB method
(2) PCR amplification
A. reaction system: 10 μ L systems, the content of each component materials is respectively 15ng template DNA；5pmol·L-1Primer；
1.0mmol·L-1dNTP；1.0uL 10×PCR Buffer；0.5U Taq archaeal dna polymerase；ddH2O polishing 10 μ L, mixing, from
The heart,
Wherein the sequence of primer is F TCR540:5 '-TGACATTGTTGATGTGCTGA-3 ',
R TCR540:5’-AAATATGCCTTCAATTGCTTC-3’；
B. amplification program: 94 DEG C of denaturations 5min；94 DEG C of degeneration 30s；60 DEG C of annealing 45s；72 DEG C extend 30s；35 are followed
After ring；72 DEG C extend 10min；Last 4 DEG C of preservations；
C. electrophoresis: mixed with isopyknic sample-loading buffer by amplified production, 94 DEG C of degeneration 6min, afterwards at DNA sequencing
6% denaturing polyacrylamide gel electrophoresis is carried out on electrophresis apparatus, electrophoresis 1.5 hours under the conditions of 70W, after electrophoresis terminates, carry out silver
Dye, and observe amplification；
D. judge: if able to 148bp detected and more than 148bp two band in amplification, then detected materials contains
The probability having Chinese cabbage anti-clubroot gene C Rb is 100%.
The present invention provide a kind of Chinese cabbage anti-clubroot CRb gene be divided into from molecular marker TCR540, with Vegetable Crops of Brassica
Anti-clubroot gene C Rb be divided into from, the molecular marker assisted selection that can be widely applied to Vegetable Crops of Brassica anti-clubroot gene C Rb is educated
Kind.The present invention can carry out seedling stage assay to plant to be measured by specific primer PCR amplification, is greatly improved breeding efficiency, and shortening is educated
The process of kind.
Accompanying drawing explanation
Fig. 1 is that TCR540 and TCR541 is in disease-resistant material ' CRBJN3-2 ', susceptible material ' BJN3-2 ' and recombinant
Amplification；
Wherein, Figure 1A is that TCR540 is at disease-resistant material ' CRBJN3-2 ' (R swimming lane), susceptible material ' BJN3-2 ' (S swimming lane)
And the amplification in recombinant (1-44 swimming lane), Figure 1B is that TCR541 is in disease-resistant material ' CRBJN3-2 ' (R swimming lane), sense
Amplification in sick material ' BJN3-2 ' (S swimming lane) and recombinant (1-44 swimming lane)；
Fig. 2 is alignment's figure that the primer of TCR540 expands out in the middle of the disease-resistant material of Chinese cabbage and susceptible material；
Fig. 3 is the genetic linkage maps of Chinese cabbage anti-clubroot CRb gene.
Detailed description of the invention
The present invention is described in detail with detailed description of the invention below in conjunction with the accompanying drawings, it is to be understood that the protection of the present invention
Scope is not limited by detailed description of the invention.
One, Chinese cabbage anti-clubroot CRb gene be divided into the acquisition from molecular marker TCR540 and TCR541
1, the structure of segregating population
With the Chinese cabbage strain ' CRBJN3-2 ' containing anti-clubroot gene C Rb as male parent, the Chinese cabbage of easy infection clubroot
Strain ' BJN3-2 ' is that hybridization of female parent obtains F1, obtained F1All show as disease-resistant for plant.Two above parent ' CRBJN3-2 '
' BJN3-2 ' is all stored in Agricultural University Of Shenyang.Select individual plant F1Selfing builds F2For mapping population, colony's number is 4783.
Whole 4783 F of sowing2Colony, inoculates plasmodiophora brassicae after planting 10 days.Disease Resistance Identification, 4783 F are carried out after 40 days2Colony
In 3641 disease-resistant, 1142 are susceptible, meet the segregation ratio of 3:1 through X 2 test
2, CTAB method extracts DNA
1), in 1.5ml centrifuge tube, add 498uL 1 × extract with CTAB liquid and 2uL beta-mercaptoethanol, shake up, wherein 1
× extract with CTAB liquid comprises the CTAB of 2%, the NaCl of EDTA and 1.4mol/L of the Tris-HCl of 100mmol/L, 20mmol/L,
The mass concentration that wherein CTAB of 2% refers to is 2% (w/v)；
2), take 0.2g young leaflet tablet, under the conditions of liquid nitrogen, be ground to powder, join containing extract with CTAB liquid and β-sulfydryl
In the centrifuge tube of ethanol, shake up；
3), put in 65 DEG C of water-baths subsequently by centrifuge tube, shook once every 5 minutes, water-bath 30min；
4), take out centrifuge tube, add isopyknic chloroform: iso pentane alcohol mixture (24:1, v/v), after rocking 10min, often
The lower 12000r/min of temperature is centrifuged 10min；
5), by supernatant transfer in another centrifuge tube, add isopyknic chloroform: isoamyl alcohol (24:1, v/v), rock
After 10min, under room temperature, 12000r/min is centrifuged 10min；
6), by supernatant move in the middle of another centrifuge tube, add the dehydrated alcohol of 2 times of prior pre-coolings of volume, after mixing, will
The DNA united chooses in the centrifuge tube equipped with 400uLTE buffer and dissolves；
7), add 1.5uLRNaseA (10ug/uL), mix latter 37 DEG C and preserve 30min；
8), step 5 is repeated)；
9), by supernatant transfer in another centrifuge tube, in centrifuge tube, add 50uL 3mol/LNaAC solution and pre-cooling
Isopyknic isopropanol ,-20 DEG C precipitation 20min；
10), 12000r/m is centrifuged 10min under the conditions of 4 DEG C, discards supernatant, adds the ethanol purge 2 times of 75%；
11), on superclean bench, air-dry DNA, add 50 μ L TE (Tris-EDTA) dissolving DNAs ,-20 DEG C of refrigerators are protected
Deposit.
Two, Chinese cabbage anti-clubroot CRb gene be divided into the acquisition from molecular marker TCR540 and TCR541 and qualification
1, the screening of polymorphism primer
2 flanking marker TCR79 of the anti-clubroot CRb gene delivered for 2014 according to Piao Zhong cloud seminar and
TCR108 sequence (Zhang etc., Fine genetic and physical mapping ofthe CRb gene
Conferring resistance to clubroot disease in Brassica rapa., Mol Breeding, 34:
1173-1183), this gene is anchored in Chinese cabbage A3 linkage group 83.5kb interval.Analyze Brassica Database data
Storehouse (http://brassicadb.org) sequence information, finds that target area exists 15 genes.Base according to these 15 genes
Because of function, wherein 5 disease-resistant related genes are checked order on disease-resistant material ' CRBJN3-2 '.Sequencing result is compared
Right, find disease-resistant material has the sequence of 3 genes and Brassica Database data base (http: //
Brassicadb.org) there are differences with reference to genome sequence in.Therefore, according to the sequencing result of disease-resistant material, utilize
Primer5.0 software, design 3 is to primer altogether.
Utilize 3 pairs of primer amplification parents ' CRBJN3-2 ' and the DNA of ' BJN3-2 ' of design, wherein have 2 pairs of primers parent
This has polymorphism, and is respectively designated as the sequence of molecular marker corresponding to TCR540 and TCR541, TCR540 and TCR541
It is respectively as follows:
(1) nucleotide sequence of TCR540 (148bp) is as shown in SEQ ID NO.1, for:
5’-TGACATTGTTGATGTGCTGATAAAAAAATCAGTAAGTCTAAGCAAATGTATATATTTTAGTTCTAT
TGGAACTTTGGATTACTGCTTTCTTCGCTTAGAAAAGTTTGCATTTTTACTTCAGGGAACCGAAGCAATTGAAGGCA
TATTT-3’。
(2) nucleotide sequence of TCR541 (252bp) is as shown in SEQ ID NO.4, for:
5'-TGCTTGAGCAGAAACAATATCAAAAATCTACCTGGAAGCATCAAGAAACTTCATCATCTCAAATCT
CTTTACTTGCATTGTCAACAGCTCGTTTCTCTTCCACTGCTTCCATCAAATCTGCAGTATCTGGATGCTCATGGCTG
TATCTCACTCGAAACAGTGGCTAAACCCATGACGCTTCTTGTGGTAGCTGAAAGGAACCAGTCTACTTTCGTCTTCA
CTGATTGTTTCAAGCTAAACAGAGATGCGCAA-3’。
Separately design according to above-mentioned sequence and synthesized the pair of primers F TCR540/R TCR540 for TCR540, with
And the pair of primers F TCR541/R TCR541 for TCR541, particular sequence is as follows:
(1) primer of TCR540:
F TCR540:5 '-TGACATTGTTGATGTGCTGA-3 ' (nucleotide sequence is as shown in SEQ ID NO.2)
R TCR540:5 '-AAATATGCCTTCAATTGCTTC-3 ' (nucleotide sequence is as shown in SEQ ID NO.3).
(2) primer of TCR541:
F TCR541:5-TGCTTGAGCAGAAACAATATCAA-3 ' (nucleotide sequence is as shown in SEQ ID NO.5)
R TCR541:5 '-TTGCGCATCTCTGTTTAGCTT-3 ' (nucleotide sequence is as shown in SEQ ID NO.6).
2, the qualification of molecular marker
In order to verify the reliability of these molecular markers, first, CRb both wings linked marker TCR79 and TCR74 is utilized to see
(Zhang etc., Fine genetic and physical mapping of the CRb gene conferring
Resistance to clubroot disease in Brassica rapa., Mol Breeding, 34:1173-1183) sieve
Select 1142 strain F2For the recombinant in susceptible individual；Then, utilize TCR540 primer (F TCR540/R TCR540) and
The primer (F TCR541/RTCR541) of TCR541 is respectively to Chinese cabbage ' the CR BJN3-2 ' and easily containing anti-clubroot gene C Rb
The Chinese cabbage inbred lines ' BJN3-2 ' of sense clubroot and 44 F thereof2PCR amplification has been carried out for susceptible recombinant.Amplification table
It is bright as it is shown in figure 1, Figure 1A is that TCR540 is at disease-resistant material ' CR BJN3-2 ' (R swimming lane), susceptible material ' BJN3-2 ' (S swimming lane)
And the amplification in recombinant (1-44 swimming lane), use the primer of TCR540 to expand, at disease-resistant parent ' CR
The band of a 148bp can be amplified, at Susceptible parent ' BJN3-2 ' and 44 strain F on BJN3-2 '2For energy equal on susceptible recombinant
Amplify a band more than 148bp；Figure 1B is that TCR541 is at disease-resistant material ' CR BJN3-2 ' (R swimming lane), susceptible material
Amplification in ' BJN3-2 ' (S swimming lane) and recombinant (1-44 swimming lane), uses the primer of TCR541 to expand,
The band of a 252bp can be amplified, at Susceptible parent ' BJN3-2 ' and 44 strain F on disease-resistant parent ' CRBJN3-2 '2For susceptible
A band more than 252bp all can be amplified on recombinant.
The sequence that the primer of TCR540 expands out in the middle of disease-resistant material is:
5’-TGACATTGTTGATGTGCTGATAAAAAAATCAGTAAGTCTAAGCAAATGTATATATTTTAGTTCTAT
TGGAACTTTGGATTACTGCTTTCTTCGCTTAGAAAAGTTTGCATTTTTACTTCAGGGAACCGAAGCAATTGAAGGCA
TATTT-3’。
The sequence that the primer of TCR540 expands out in the middle of susceptible material is:
5’-TGACATTGTTGATGTGCTGACCAACAACTCAGTAAGTCAGAGAAAATGTATATCTGTTACTTCCAC
TGCTAGAAAATGTATATCTATTACTTCCACTGAAATTTAGAATACTGTTTAGGTTCCTTTGTACACTTAGAAGCTTA
TGTTTCTGAACTCATTTGCCATTTTTCTTCAGGGAACAGAAGCAATTGAAGGCATATTT-3’。
The comparing result of above-mentioned two sequence is as in figure 2 it is shown, result shows, two sequences exist bigger diversity, say
The molecular marker of bright TCR540 can be good at distinguishing the disease-resistant material of Chinese cabbage and susceptible material, can be used for Vegetable Crops of Brassica and includes balling
The molecular marker auxiliary choosing of Chinese cabbage and green stem vegetable anti-clubroot gene.
3, the structure of genetic map
According to F2The Screening and Identification result of colony, calculates each labelling and the intergenic recombination fraction of CRb, and Mapchart2.1 paints
The genetic linkage maps (Fig. 3) of CRb gene processed, on the right side of figure, the numeral between labelling represents the recombinant number between labelling, on the left of figure
Numeral represent the genetic distance between two labellings, TCR79, TCR108, TCR30, TCR37, TCR74 as seen from the figure, fixed respectively
It is positioned at distance CRb gene 0.03cM, 0.04cM, 0.36cM, 0.43cM, 0.49cM.And molecular marker TCR540 newly developed,
There is separation relation altogether in TCR541 Yu CRb gene.Accordingly, it is determined that these 2 labellings can apply to the anti-clubroot of Chinese cabbage from now on
In the middle of the molecular marking supplementary breeding process of CRb gene, and owing to these 2 molecular markers are altogether separation marking, with before
Linked marker is compared, and can eliminate Linkage drag phenomenon, be greatly improved transformation efficiency in the application of molecular mark.
Three, being divided into from molecular marker TCR540 and TCR541 in assisted Selection DABAI of Chinese cabbage anti-clubroot CRb gene
Application process in dish anti-clubroot plant, specifically includes:
1, the genomic DNA of detected materials is extracted by CTAB method
With CTAB method extract Chinese cabbage material to be measured genomic DNA, concrete steps with reference to above " one, Chinese cabbage
Anti-clubroot CRb gene be divided into the acquisition from molecular marker TCR540, TCR541 " in " 2, CTAB method extract DNA " part note
The content carried.
2, PCR amplification
A. reaction system: 10 μ L systems, the content of each component materials is respectively 15ng template DNA；5pmol·L-1Primer；
1.0mmol·L-1dNTP；1.0uL 10×PCR Buffer；0.5U Taq archaeal dna polymerase；ddH2O polishing 10 μ L, mixing, from
The heart,
Wherein, the sequence of the primer that TCR540 uses is:
F TCR540:5 '-TGACATTGTTGATGTGCTGA-3 ',
R TCR540:5 '-AAATATGCCTTCAATTGCTTC-3 ',
The sequence of the primer that TCR541 uses is:
F TCR541:5 '-TGCTTGAGCAGAAACAATATCAA-3 ',
R TCR541:5’-TTGCGCATCTCTGTTTAGCTT-3’；
B. amplification program: 94 DEG C of denaturations 5min；94 DEG C of degeneration 30s；60 DEG C of annealing 45s；72 DEG C extend 30s；35 are followed
After ring；72 DEG C extend 10min；Last 4 DEG C of preservations.
3, electrophoresis:
TCR540: mixed with isopyknic sample-loading buffer by amplified production, 94 DEG C of degeneration 6min, afterwards at DNA sequencing
6% denaturing polyacrylamide gel electrophoresis is carried out on electrophresis apparatus, electrophoresis 1.5 hours under the conditions of 70W, after electrophoresis terminates, carry out silver
Dye；
TCR541: mixed with isopyknic sample-loading buffer by amplified production, 94 DEG C of degeneration 6min, afterwards at DNA sequencing
6% denaturing polyacrylamide gel electrophoresis is carried out on electrophresis apparatus, electrophoresis 2 hours under the conditions of 70W, after electrophoresis terminates, carry out silver staining.
It should be noted that sample-loading buffer uses the buffer that polyacrylamide gel electrophoresis is conventional, wherein comprise
The xylene cyanol's and 0.001% (w/v) of the EDTA of the formamide, 10mM of 98% (w/v), 0.001% (w/v)
bromphenol blue。
4, interpretation of result
1. if able to 148bp detected and more than 148bp two band in using the amplification of primer of TCR540,
The probability then containing Chinese cabbage anti-clubroot gene C Rb in detected materials is 100%.
2. if able to 252bp detected and more than 252bp two band in using the amplification of primer of TCR541,
The probability then containing Chinese cabbage anti-clubroot gene C Rb in detected materials is 100%.
If 1., 2. the amplification in 2 all can obtain, then detected materials has Chinese cabbage anti-clubroot gene C Rb
Probability be 100%.
Although preferred embodiments of the present invention have been described, but those skilled in the art once know basic creation
Property concept, then can make other change and amendment to these embodiments.So, claims are intended to be construed to include excellent
Select embodiment and fall into all changes and the amendment of the scope of the invention.
Obviously, those skilled in the art can carry out various change and the modification essence without deviating from the present invention to the present invention
God and scope.So, if these amendments of the present invention and modification belong to the scope of the claims in the present invention and equivalent technologies thereof
Within, then the present invention is also intended to comprise these change and modification.
Claims (5)
1. a Chinese cabbage anti-clubroot CRb gene be divided into from molecular marker TCR540, it is characterised in that described Chinese cabbage resist
Clubroot CRb gene be divided into the nucleotide sequence from molecular marker TCR540 as shown in SEQ ID NO.1, for:
5’-TGACATTGTTGATGTGCTGATAAAAAAATCAGTAAGTCTAAGCAAATGTATATATTTTAGTTCTATTGGA
ACTTTGGATTACTGCTTTCTTCGCTTAGAAAAGTTTGCATTTTTACTTCAGGGAACCGAAGCAATTGAAGGCATATT
T-3’。
2. the Chinese cabbage anti-clubroot CRb gene described in a claim 1 to be divided into the PCR from molecular marker TCR540 special
Property amplimer includes:
F TCR540:5’-TGACATTGTTGATGTGCTGA-3’
R TCR540:5’-AAATATGCCTTCAATTGCTTC-3’。
Chinese cabbage the most according to claim 1 anti-clubroot CRb gene be divided into from molecular marker TCR540, at molecule mark
Application in note assisted Selection.
Chinese cabbage the most according to claim 1 anti-clubroot CRb gene be divided into from molecular marker TCR540, in Chinese cabbage
Application in anti-clubroot molecular marker assisted selection.
5. the Chinese cabbage anti-clubroot CRb gene described in a claim 1 be divided into the application side from molecular marker TCR540
Method, specifically includes:
(1) genomic DNA of detected materials is extracted by CTAB method
(2) PCR amplification
A. reaction system: 10 μ L systems, the content of each component materials is respectively 15ng template DNA；5pmol·L-1Primer；
1.0mmol·L-1dNTP；1.0uL 10×PCR Buffer；0.5U Taq archaeal dna polymerase；ddH2O polishing 10 μ L, mixing, from
The heart,
Wherein the sequence of primer is F TCR540:5 '-TGACATTGTTGATGTGCTGA-3 ',
R TCR540:5’-AAATATGCCTTCAATTGCTTC-3’；
B. amplification program: 94 DEG C of denaturations 5min；94 DEG C of degeneration 30s；60 DEG C of annealing 45s；72 DEG C extend 30s；After 35 circulations；
72 DEG C extend 10min；Last 4 DEG C of preservations；
C. electrophoresis: mixed with isopyknic sample-loading buffer by amplified production, 94 DEG C of degeneration 6min, afterwards at DNA sequencing electrophoresis
6% denaturing polyacrylamide gel electrophoresis is carried out on instrument, electrophoresis 1.5 hours under the conditions of 70W, after electrophoresis terminates, carry out silver staining, and
Observe amplification；
D. judge: if able to 148bp detected and more than 148bp two band in amplification, then containing big in detected materials
The probability of Chinese cabbage anti-clubroot gene C Rb is 100%.
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| CN108588264A (en) * | 2018-06-08 | 2018-09-28 | 西南大学 | Molecular labeling, primer and its application for identifying club-root resistance |
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| CN110734999A (en) * | 2019-11-12 | 2020-01-31 | 北京市农林科学院 | SNP molecular marker tightly linked with new clubroot-resistant gene CRw of Chinese cabbage and application thereof |
| CN112553359A (en) * | 2021-02-04 | 2021-03-26 | 沈阳农业大学 | Clubroot molecular marker syau3008 co-separated from Chinese cabbage gene, primer and application |
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| CN108285930A (en) * | 2018-04-19 | 2018-07-17 | 青岛市农业科学研究院 | With SSR molecular marker, kit and its application of the anti-clubroot character close linkage of the high CR117 Chinese cabbages of moral |
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| CN108588264A (en) * | 2018-06-08 | 2018-09-28 | 西南大学 | Molecular labeling, primer and its application for identifying club-root resistance |
| CN109295247A (en) * | 2018-10-22 | 2019-02-01 | 沈阳农业大学 | Compact linkage molecule label yau403, primer and the application of the anti-clubroot CRd gene of Chinese cabbage |
| CN109295247B (en) * | 2018-10-22 | 2021-11-23 | 沈阳农业大学 | Closely linked molecular marker yau403 of clubroot-resistant CRd gene of Chinese cabbage, primer and application |
| CN110734999B (en) * | 2019-11-12 | 2020-06-16 | 北京市农林科学院 | SNP molecular marker tightly linked with new clubroot-resistant gene CRw of Chinese cabbage and application thereof |
| CN110734999A (en) * | 2019-11-12 | 2020-01-31 | 北京市农林科学院 | SNP molecular marker tightly linked with new clubroot-resistant gene CRw of Chinese cabbage and application thereof |
| CN112553359A (en) * | 2021-02-04 | 2021-03-26 | 沈阳农业大学 | Clubroot molecular marker syau3008 co-separated from Chinese cabbage gene, primer and application |
| CN112553359B (en) * | 2021-02-04 | 2023-05-05 | 沈阳农业大学 | Clubroot molecular marker syau3008 coseparated from Chinese cabbage genes, primer and application |
| CN114381463A (en) * | 2022-02-23 | 2022-04-22 | 华中农业大学 | Application of BraA08g039212E gene from European turnip ECD04 in improvement of plasmodiophora tumefaciens resistance |
| CN114381463B (en) * | 2022-02-23 | 2023-11-07 | 华中农业大学 | Application of BraA08g039212E gene derived from European turnip ECD04 in improving resistance of rhizopus |
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