CN108546775A - The InDel labels and its detection primer of a kind of Chinese cabbage burr and application - Google Patents
The InDel labels and its detection primer of a kind of Chinese cabbage burr and application Download PDFInfo
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- CN108546775A CN108546775A CN201810725802.7A CN201810725802A CN108546775A CN 108546775 A CN108546775 A CN 108546775A CN 201810725802 A CN201810725802 A CN 201810725802A CN 108546775 A CN108546775 A CN 108546775A
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Abstract
The present invention relates to the detection primer of InDel labels and its applications in impulse- free robustness Chinese cabbage breeding.The detection primer of Chinese cabbage InDel labels TR 1, which is a pair, and sense primer nucleotide sequence is as shown in SEQ ID NO.1, and downstream primer nucleotide sequence is as shown in SEQ ID NO.2.Present invention firstly discloses mark TR 1 and its detection primer with the relevant InDel of Chinese cabbage burr, by carrying out PCR amplification for the DNA of segregating population material to the F2 generated after hairiness and hairless Chinese cabbage inbred lines and the two hybridization, and it is detected using 8% modacrylic acyl ammonia gel electrophoresis, it was found that can obviously distinguish hairiness and hairless Chinese cabbage cultivar with the relevant InDel labels TR 1 of Chinese cabbage burr, it can be used for the selection and breeding of hairless New Chinese Cabbage Variety.
Description
Technical field
The present invention relates to a kind of InDel of Chinese cabbage burr labels and its detection primer and applications, belong to molecular biology
Technical field.
Background technology
Molecular marker assisted selection (marker-assisted selection, MAS) is not limited by environmental condition, can
To carry out (Tanksley, 1989) from DNA molecular level in any growth period of development of plants.It overcomes traditional plant
Breeding time is long, speed is slow, uncertain, the defects of being limited by environmental condition.Currently, various plants point have had been established in the world
Sub- labelling technique system and the genetic improvement for being applied to crops, this opens new approach for crop breeding.Utilize molecule mark
Note technology quickly screens the appearance of especially molecular labeling so that the selection in breeding work is changed by traditional Phenotypic Selection
Genotype selects, and substantially increases the accuracy and efficiency of selection.The molecular labeling of based on PCR such as RAPD (random
amplified polymorphic DNA)、SSR(Simple sequence repeat)、AFLP(amplified
Fragment length polymorphism), the appearance of InDel (insertion-deletion) etc., because it is easy, quickly
And the features such as efficient, so that the development and utilization of molecular labeling is spread various crops.Wherein InDel labels are due at same site
DNA sequence dna the insertion/deletions of nucleotide fragments has occurred between Different Individual, it is special according to the sequence design of target site both sides
Different primer carries out PCR amplification, and the length polymorphism of amplified fragments is InDel labels.Since InDel is labeled as codominance mark
Note, has many advantages, such as preferable stability and rich polymorphism, has been widely used in the finely positioning of polymorphism analysis, gene
And map based cloning etc..
Chinese cabbage (Brassica rapa ssp.pekinensis, AA genomes, 2n=20) is China originating from China
One of cultivated area and the maximum vegetable crop of yield have important economic value, are known as the title of " state's dish ".Chinese cabbage leaf quilt
Hair is a kind of protective tissue on plant surface, is the outermost layer that plant contacts with environment.It has the infringement of prevention disease pest and purple
Outside line injures, the effects that reducing plant transpiration effect and increase freexing tolerance, but as the improvement of people's living standards, because of its mouth
Adaptive and sense of touch, most consumers more have a preference for hairless Chinese cabbage cultivar, and therefore, whether there is or not leaf hair, to be that current Chinese cabbage is disease-resistant educate
Kind, the important goal character of quality breeding and commodity property.But up to the present, more with the label of burr character close linkage
Lack, seriously hinders the process of its breeding, it means that the molecular labeling of exploitation and Chinese cabbage burr character close linkage, it is right
To there are important guidance and practice significance in the hairless disease-resistant New Chinese Cabbage Variety of quickly breeding.
Invention content
In view of the deficiencies of the prior art, the present invention provides a kind of InDel of Chinese cabbage burr label and its detection primer with
Using.
Technical solution of the present invention is as follows：
A kind of InDel label TR-1 of Chinese cabbage burr, nucleotide sequence such as SEQ ID NO.3 or SEQ ID NO.4 institutes
Show.
The detection primer of the InDel labels TR-1 of above-mentioned Chinese cabbage burr, which is a pair, sense primer nucleosides
Acid sequence is as shown in SEQ ID NO.1, and downstream primer nucleotide sequence is as shown in SEQ ID NO.2.
Application of the detection primer of the InDel labels TR-1 of above-mentioned Chinese cabbage burr in hairless Chinese Cabbage.
According to currently preferred, the application, steps are as follows：
(1) genomic DNA for extracting Chinese cabbage cultivar to be detected, obtains Genomic DNA solution；
(2) using genomic DNA made from step (1) as template, PCR is carried out to genomic DNA using above-mentioned detection primer
Pcr amplification product is made in amplification；
(3) by pcr amplification product made from step (2) after gel electrophoresis, when PCR product electrophoresis pattern shows detected sample
When product only have mono- band of 156bp, then the tested sample is to have batt material；When PCR product electrophoresis pattern shows only 144bp
When one band, then the tested sample is no batt material；Contain 156bp and 144bp simultaneously when PCR product electrophoresis pattern is shown
When band, then the tested sample is hairiness and hairless cross materials, and batt material has been shown as in phenotype.
According to the present invention it is further preferred that in the step (1), the genomic DNA of Chinese cabbage cultivar to be detected is extracted
It is extracted using the CTAB methods of improvement.
According to the present invention it is further preferred that in the step (2), PCR reaction systems are as follows, and total system is 20 μ L：
20ng/ μ L template DNAs 2 μ L, MgCl containing 20mM210 × PCR buffer2 μ L, 2.5mMdNTPs0.5 μ L, 10
μM sense primer 0.4 μ L, 10 μM of 0.4 μ L, 1U Taq DNA polymerase of downstream primer, 0.2 μ L add ddH2O to 20 μ L.
According to the present invention it is further preferred that in the step (2), PCR amplification program is：
95 DEG C of pre-degeneration 5min；94 DEG C of denaturation 30s, 58 DEG C of annealing 30s, 72 DEG C of extension 30s amount to 36 cycles；72℃
Extend 10min.
According to the present invention it is further preferred that in the step (3), gel electrophoresis is the denaturation that mass percent is 8%
Polyacrylamide gel electrophoresis.
Advantageous effect
Present invention firstly discloses TR-1 and its detection primer is marked with the relevant InDel of Chinese cabbage burr, by having
The F2 generated after hair and hairless Chinese cabbage inbred lines and the two hybridization carries out PCR amplification, and profit for the DNA of segregating population material
It is detected with 8% modacrylic acyl ammonia gel electrophoresis, finding can be apparent with the relevant InDel labels TR-1 of Chinese cabbage burr
Hairiness and hairless Chinese cabbage cultivar are distinguished, the selection and breeding of hairless New Chinese Cabbage Variety are can be used for.
Description of the drawings
Fig. 1 is nucleotide alignments' figure of the relevant InDel labels TR-1 of Chinese cabbage burr；
Identified areas of crossing in figure is that Indel marks TR-1；Hairiness Chinese cabbage (G291) and hairless Chinese cabbage (ZHB) phase
Than there is the DNA of 12bp to be inserted into；
Fig. 2 is that the detection primer of the relevant InDel labels TR-1 of Chinese cabbage burr is big in hairless (ZHB) and hairiness (G291)
The F2 generated after Chinese cabbage and the two hybridization is for the PCR amplification electrophoresis result photo in segregating population material；
In figure：Swimming lane 1：ZHB (hairless), swimming lane 2：G291 (hairiness), swimming lane 3-14：ZHB and G291 hybridizes F2 for hairless
Group, swimming lane 15-26：ZHB and G291 hybridizes F2 for hairiness group；
Fig. 3 be the hairiness sold on the market of the detection primer of Chinese cabbage burr relevant InDel label TR-1 with it is hairless
PCR amplification electrophoresis result photo in Chinese cabbage material；
In figure：Swimming lane 1：ZHB (hairless), swimming lane 2：G291 (hairiness), swimming lane 3-17 (hairless)：It is AM4-1, AM3- respectively
1, AM5-2, AM9-1, AM94, AM30, AM54, AM72-1, AM10-1, AM56-1, blueness grind No. 2 white spring, the high lily of moral, west in vain
45, Y2, L41, swimming lane 18-32 (hairiness)：Western white No. 8 respectively, China 1, blueness grind CR20, blueness grind the summer is No. 2 white, Guangdong is early,
L46,2013-33, AM2-1, highly resistance 2, white No. 5 western, Xia You bis-, golden boy and jade girl, tackling key problem two, rich anti-60, moral Gao Xikang
65。
Fig. 4 be Chinese cabbage burr relevant InDel label TR-1 detection primer with using same procedure design other 7
To InDel labeled primers to the PCR amplification electrophoresis result photo in hairiness and hairless Chinese cabbage material；
In figure：Swimming lane 1-4 (hairless)：It is ZHB, AM3-1, the high lily of moral, AM10-1, swimming lane 5-8 (hairiness) respectively：Respectively
It is G291, China 1, white No. 8 western, tackling key problem two；A1-A7 is the ineffective InDel labels of screening process respectively in figure.
Specific implementation mode
Technical scheme of the present invention is further elaborated with reference to embodiment and Figure of description, but the present invention is protected
It is without being limited thereto to protect range.
Biological material source
AM4-1、AM3-1、AM5-2、AM9-1、AM94、AM30、AM54、AM72-1、AM10-1、AM56-1、Y2、L41、
No. two AM2-1, Guangdong morning, L46, highly resistance 2,2013-33, tackling key problem kinds are purchased from Shandong Academy of Agricultural Sciences；
The high lily of moral, the happiness of moral height resist 65 kinds to be purchased from Dezhou City Shandong Province De Gao vegetable seedlings research institute；
Western white 45, western white No. 8 kinds are purchased from Shandong Denghai Seeds Co., Ltd；
No. a kind of China is purchased from Shandong Province Linqu County De Minzhong industry Co., Ltd；
Blueness grinds CR20, blueness grinds that the summer is No. 2 white, blueness grinds spring white No. 2 kinds and is purchased from Qingdao of Shandong province academy of agricultural science；
Western white No. 5, rich anti-60 kinds be purchased from Shandong Province Laizhou Institute of Agricultural Science vegetable seedling research institute；
Summer excellent No. two kinds are purchased from Licheng City spring and summer Chinese cabbage research institute；
Golden boy and jade girl's kind is purchased from Beijing Ao Shuofeng agricultural science and technologys Co., Ltd.
The exploitation of embodiment 1.InDel labeled primers
Using Chinese cabbage burr formation related gene as candidate gene, using weight sequencing technologies to hairiness (G291) with it is hairless
(ZHB) Chinese cabbage carries out full-length genome and resurveys sequence.According to weight sequencing result to the candidate gene in hairiness and hairless Chinese cabbage into
Row sequence alignment marks TR-1 design primers to 1 InDel therein, the sequence alignment figure of Chinese cabbage InDel labels TR-1
As shown in Figure 1.Identified areas of crossing in figure is that InDel marks TR-1.
Hairiness Chinese cabbage (G291) has the DNA of 12bp to be inserted into compared with hairless Chinese cabbage (ZHB).Design Chinese cabbage InDel
The detection primer of TR-1 is marked, primer sequence is as shown in SEQ ID NO.1 and SEQ ID NO.2.
The application of embodiment 2.InDel labeled primers
The extraction of 2.1 vegetable materials and template DNA
It chooses Chinese cabbage first generation of hybrid commercial variety blueness and grinds No. 2 white spring, the high lily of moral, western white 45, white No. 8 western, China 1
Number, blueness grind CR20, blueness grind the summer it is No. 2 white, China No. 1, highly resistance 2, white No. 5 western, Xia You bis-, golden boy and jade girl, public relations No. two, it is rich
Anti- 60, Huayang is white, moral is high likes anti-65 and Chinese cabbage inbred lines Guangdong morning, Y2, L41, L46, AM4-1, AM3-1, AM5-2, AM9-
1, the hybridization F2 of AM94, AM30, AM54, AM72-1, AM10-1, AM56-1 and ZHB and G291 are material for blade.
Total DNA is extracted with the CTAB methods of improvement, is as follows：
1. taking the fresh cabbage leaves of 1-10g, powder is quickly ground into liquid nitrogen；
2. agar is transferred in the centrifuge tube of precooling, isometric (w/v, the g/ml) 2 × CTAB Extraction buffers of addition, 65 DEG C
Heat preservation 20 minutes；
3. isometric chloroform/isoamyl alcohol (volume ratio 24 is added:1), overturn centrifuge tube mixing, 12000r/min room temperatures from
The heart 10 minutes；
4. supernatant is transferred in another centrifuge tube, isometric chloroform/isoamyl alcohol (volume ratio 24 is added:1), overturn from
Heart pipe mixing, 12000r/min room temperatures centrifuge 10 minutes；
5. upper strata aqueous phase is transferred in new centrifuge tube, isometric isopropanol is added, gently overturns mixing, room temperature decentralization
It sets 30 minutes；
6.12000r/min room temperatures centrifuge 5-10 minutes, remove supernatant；
7.70% ethyl alcohol rinses, and 12000r/min room temperatures centrifuge 5-10 minutes, remove supernatant；
8. it is primary to repeat step 7；
9. precipitation air-dries, the TE buffer solutions or ultra-pure water dissolving DNA of 100 μ l is added, -20 DEG C save backup；
10. 2 μ l solution is taken to be detected using 1% agarose gel electrophoresis.
2.2PCR amplification
Detection primer (the SEQ ID NO.1 and SEQ ID NO.2 of TR-1 are marked according to the Chinese cabbage InDel of above-mentioned design
Nucleotide sequence), mark the primer of TR-1 in Sangon Biotech (Shanghai) Co., Ltd. synthesis Chinese cabbage InDel,
Be diluted to after dissolving 10 μM it is spare.
Using the DNA of said extracted as template, PCR amplification is carried out with the primer of Chinese cabbage InDel labels respectively, PCR is made
Product.
PCR reaction systems are as follows, 20 μ L of total system：
20ng/ μ L template DNAs 2 μ L, the MgCl containing 20mM210 × PCR buffer2 μ L, concentration 2.5mM's
DNTPs0.5 μ L, the 0.4 μ L of sense primer that 10 μM of concentration, the Taq DNA polymerase of downstream primer 0.4 the μ L, 1U that 10 μM of concentration
0.2 μ L, add ddH2O to 20 μ L.
Amplification program is：
95 DEG C of pre-degeneration 5min；94 DEG C of denaturation 30s, 57 DEG C of annealing 30s, 72 DEG C of extension 30s amount to 36 cycles；72℃
Extend 10min.
2.3 polyacrylamid gels are separated by electrophoresis and polymorphic detection
PCR product is developed using argentation and is detected after mass concentration is 8% modacrylic acyl ammonia gel electrophoresis.
PCR product and 95 DEG C of pre-degeneration 10min after sample-loading buffer mixing, take 4 μ l point samples, in Sequi-GT cores
It is separated by electrophoresis in sour electrophoresis system (Bio-Rad, USA), electrode buffer is 1 × TBE, electric under 25 DEG C, 55W invariable powers
Swim 1.5h, by argentation development, takes pictures and records result.
Fig. 2 is primer pair ZHB, G291 and ZHB and G291 hybridization F2 groups material using Chinese cabbage InDel labels TR-1
The pcr amplification product of material carries out the result figure after 8% modacrylic acyl ammonia gel electrophoresis.
Fig. 3 be using Chinese cabbage InDel label TR-1 primer pair without batt material (AM4-1, AM3-1, AM5-2, AM9-1,
AM94, AM30, AM54, AM72-1, AM10-1, AM56-1, blueness grind No. 2 white spring, the high lily of moral, western white 45, Y2, L41) and hairiness
Material (western white No. 8, China 1, blueness grinds CR20, blueness grinds No. 2 white summer, Guangdong morning, L46, No. 1 Chinese, highly resistance 2, it is white No. 5 western,
Summer is No. two excellent, golden boy and jade girl, public relations two, rich anti-60, Huayang is white, the high anti-pcr amplification product 65) of happiness of moral carries out mass concentration
For the result figure after 8% modacrylic acyl ammonia gel electrophoresis.
Fig. 4 is using Chinese cabbage InDel label TR-1 primers and using other 7 pairs of InDel labels of same procedure design
Primer pair without batt material (early emperor is white, AM3-1, the high lily of moral, AM10-1) and have batt material (hat 291, it is No. 1 Chinese, white No. 8 western,
Tackling key problem No. two) pcr amplification product carry out mass concentration be 8% modacrylic acyl ammonia gel electrophoresis after result figure.
Chinese cabbage InDel marks the primer of TR-1 can be to hairiness and hairless Chinese cabbage it can be seen from Fig. 2, Fig. 3 and Fig. 4
DNA effectively expanded, banding pattern is clear, and in hairiness and hairless Chinese cabbage material have good polymorphism, wherein having
Batt material amplified band is 156bp, and no batt material amplified band is 144bp, can be good at distinguishing hairiness and hairless Chinese cabbage,
It can be used for hairless New Chinese Cabbage Variety selection and breeding.
SEQUENCE LISTING
<110>Vegetable or flower research institute of Shandong Academy of Agricultural Sciences
<120>The InDel labels and its detection primer of a kind of Chinese cabbage burr and application
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> DNA
<213>It is artificial synthesized
<400> 1
ttccatacct ccctcactgg ta 22
<210> 2
<211> 22
<212> DNA
<213>It is artificial synthesized
<400> 2
ttcaactgtc cacaaccctt tc 22
<210> 3
<211> 144
<212> DNA
<213> ZHB
<400> 3
ttccatacct ccctcactgg tagcaatctt tataagtaga gtctctatca cagataataa 60
tctgagaatc aaaggatgag aacgaggaga agaacagagg aagagaatca tcaagaatac 120
aagaaagggt tgtggacagt tgaa 144
<210> 4
<211> 156
<212> DNA
<213> G291
<400> 4
ttccatacct ccctcactgg tagcaatctc tttctctctc tttataagta gagtctctat 60
cacagataat aatctgagaa tcaaaggatg agaacgagga gaagaacaga ggaagagaat 120
catcaagaat acaagaaagg gttgtggaca gttgaa 156
Claims (8)
1. a kind of InDel of Chinese cabbage burr marks TR-1, nucleotide sequence is as shown in SEQ ID NO.3.
2. the detection primer of the InDel labels TR-1 of Chinese cabbage burr described in claim 1, which is a pair, upstream
Primer nucleotide sequences are as shown in SEQ ID NO.1, and downstream primer nucleotide sequence is as shown in SEQ ID NO.2.
3. the detection primer of the InDel labels TR-1 of Chinese cabbage burr described in claim 2 is in hairless Chinese Cabbage
Application.
4. application as claimed in claim 3, which is characterized in that steps are as follows：
(1) genomic DNA for extracting Chinese cabbage cultivar to be detected, obtains Genomic DNA solution；
(2) using genomic DNA made from step (1) as template, PCR amplification is carried out to genomic DNA using above-mentioned detection primer,
Pcr amplification product is made；
(3) by pcr amplification product made from step (2) after gel electrophoresis, when PCR product electrophoresis pattern shows sample only
When having mono- band of 156bp, then the tested sample is to have batt material；When PCR product electrophoresis pattern shows only 144bp mono-
When band, then the tested sample is no batt material；Contain 156bp and 144bp bands simultaneously when PCR product electrophoresis pattern is shown
When, then the tested sample is hairiness and hairless cross materials, and batt material has been shown as in phenotype.
5. application as claimed in claim 4, which is characterized in that in the step (1), extract the base of Chinese cabbage cultivar to be detected
Because group DNA is using the CTAB methods extraction of improvement.
6. application as claimed in claim 4, which is characterized in that in the step (2), PCR reaction systems are as follows, and total system is
20μL：
20ng/ μ L template DNAs 2 μ L, MgCl containing 20mM210 × PCR buffer2 μ L, 2.5mMdNTPs0.5 μ L, on 10 μM
Trip primer 0.4 μ L, 10 μM of 0.4 μ L, 1U Taq DNA polymerase of downstream primer, 0.2 μ L add ddH2O to 20 μ L.
7. application as claimed in claim 4, which is characterized in that in the step (2), PCR amplification program is：
95 DEG C of pre-degeneration 5min；94 DEG C of denaturation 30s, 58 DEG C of annealing 30s, 72 DEG C of extension 30s amount to 36 cycles；72 DEG C of extensions
10min。
8. application as claimed in claim 4, which is characterized in that in the step (3), gel electrophoresis is that mass percent is
8% modacrylic acyl ammonia gel electrophoresis.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109694919A (en) * | 2018-12-29 | 2019-04-30 | 山东省农业科学院蔬菜花卉研究所 | Four seed type cabbage hybrid Purity core primers and purity of hybrid InDel molecular markers for identification kit |
| CN114317534A (en) * | 2022-03-02 | 2022-04-12 | 贵州大学 | Specific molecular marker of No.1 thorn-free Rosa roxburghii strain and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103103184A (en) * | 2013-01-28 | 2013-05-15 | 西北农林科技大学 | SSR (Simples sequence repeats) and InDel (insertion/deletion) molecular marker primer linked with brassica campestris orange head gene Br-or, and application thereof |
| CN103172715A (en) * | 2011-12-21 | 2013-06-26 | 中国科学院上海生命科学研究院 | Plant epidermal hair controlling gene and application thereof |
| CN103757018A (en) * | 2014-02-20 | 2014-04-30 | 山东省农业科学院蔬菜花卉研究所 | InDel mark relating to mutation of untranslated region of Chinese cabbage PHYB gene mRNA5'and application thereof |
| CN104774951A (en) * | 2015-04-02 | 2015-07-15 | 山东省农业科学院蔬菜花卉研究所 | Chinese cabbage EST-SSR (expressed sequence tag-simple sequence repeat) markers as well as primers and applications thereof |
| CN107164502A (en) * | 2017-06-15 | 2017-09-15 | 西南大学 | The molecular labeling being closely related and its application are whether there is with cabbage leaves epidermal hair |
-
2018
- 2018-07-04 CN CN201810725802.7A patent/CN108546775B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103172715A (en) * | 2011-12-21 | 2013-06-26 | 中国科学院上海生命科学研究院 | Plant epidermal hair controlling gene and application thereof |
| CN103103184A (en) * | 2013-01-28 | 2013-05-15 | 西北农林科技大学 | SSR (Simples sequence repeats) and InDel (insertion/deletion) molecular marker primer linked with brassica campestris orange head gene Br-or, and application thereof |
| CN103757018A (en) * | 2014-02-20 | 2014-04-30 | 山东省农业科学院蔬菜花卉研究所 | InDel mark relating to mutation of untranslated region of Chinese cabbage PHYB gene mRNA5'and application thereof |
| CN104774951A (en) * | 2015-04-02 | 2015-07-15 | 山东省农业科学院蔬菜花卉研究所 | Chinese cabbage EST-SSR (expressed sequence tag-simple sequence repeat) markers as well as primers and applications thereof |
| CN107164502A (en) * | 2017-06-15 | 2017-09-15 | 西南大学 | The molecular labeling being closely related and its application are whether there is with cabbage leaves epidermal hair |
Non-Patent Citations (1)
| Title |
|---|
| X.L. YE等: "Fine mapping and candidate gene analysis of Brtri1, a gene controlling trichome development in Chinese cabbage (Brassica rapa L. ssp pekinensis)", 《GENETICS AND MOLECULAR RESEARCH》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109694919A (en) * | 2018-12-29 | 2019-04-30 | 山东省农业科学院蔬菜花卉研究所 | Four seed type cabbage hybrid Purity core primers and purity of hybrid InDel molecular markers for identification kit |
| CN109694919B (en) * | 2018-12-29 | 2022-04-12 | 山东省农业科学院蔬菜花卉研究所 | Four-type Chinese cabbage hybrid purity identification core primer and hybrid purity InDel molecular marker identification kit |
| CN114317534A (en) * | 2022-03-02 | 2022-04-12 | 贵州大学 | Specific molecular marker of No.1 thorn-free Rosa roxburghii strain and application thereof |
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