CN104004833B - Based on EST-SSR core primers group and the application of the exploitation of sponge gourd transcript profile sequence - Google Patents
Based on EST-SSR core primers group and the application of the exploitation of sponge gourd transcript profile sequence Download PDFInfo
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Abstract
The invention discloses a set of EST-SSR core primers group based on the exploitation of sponge gourd transcript profile sequence, comprise 50 pairs of primers, is its nucleotide sequence as sequence table SEQ? ID? shown in NO.1 ~ 100.50 pairs of EST-SSR core primers of the present invention have rich polymorphism, amplification stable, reproducible, be convenient to the advantages such as statistics, simultaneously because these primers are all from sponge gourd expressing gene, the variation of functional sequence in genome can be reflected, thus better can distinguish the diversity of sponge gourd genetic germplasm.This core primers group can be used for the fields such as variety of luffa qualification, the analysis of kind Genetic lineages and Genetic Diversity of Germplasm analysis; be conducive to the legitimate rights and interests protecting breeder, producers and consumers, promote the sound development that sponge gourd genetic breeding and sponge gourd are produced.
Description
Technical field
The present invention relates to EST-SSR core primers group, be specifically related to a set of EST-SSR core primers group based on the exploitation of sponge gourd transcript profile sequence and application thereof.
Background technology
Sponge gourd is Curcurbitaceae (Cucurbitaceae) Luffa (Luffa spp.) annual climbing up by holding on to property herbaceous plant, and it all has cultivated area widely as a kind of important vegetable crop in the various places, north and south of China.Sponge gourd is except as except vegetables, or a kind of important medicinal plant.Modern pharmacology proves, containing the plurality of active ingredients such as alkaloids, flavonoid, steroid, glucosides class in sponge gourd, has the effects such as antimycotic, antibacterium, anti-inflammatory.Nearest result of study proves, containing ribosome inactivating protein in sponge gourd seed, can suppress the growth of hiv virus, be therefore the medicine of potential treatment acquired immune deficiency syndrome (AIDS).
In recent years, along with people are to the pay attention to day by day of nutritive health-care, sponge gourd is as a kind of herbal cuisine dual-purpose and high temperature season market supply vegetables, and be loved by the people, demand constantly increases, and cultivated area is expansion trend.China investigator starts the research of the aspects such as sponge gourd genetic breeding, hybrid vigour and hereditary property from the sixties in 20th century, achieves certain progress, has cultivated a collection of improved Varieties.Because the fundamental research of sponge gourd is weak, in sponge gourd breeding and kind protection, there are two outstanding problems at present: one, how precise Identification is carried out to the genetic diversity of sponge gourd germ plasm resource, thus instruct breeding man to carry out the apolegamy of parent efficiently; Two, how accurately, carry out the qualification of variety of luffa fast, thus better promote variety of luffa authorization, the development of the aspect such as kind protection, true and false kind distinguish, the solution of the property right dispute of kind.
Based on the molecular marking technique of DNA sequence polymorphism between individuality, there is the restriction in many, not affected by environment and season of test period short, potential marker number, do not have tissue specificity, accuracy high, be beneficial to the advantages such as high-throughput test analysis, be widely used in Genetic Diversity of Germplasm analysis, new variety qualification, Seed purity test.In sponge gourd, more existing investigators utilize molecule marker to carry out analysis of genetic diversity to sponge gourd germ plasm resource, but are all generally the random primer labellings such as RAPD, ISSR, the SRAP adopted.These marks are all the labeling techniques based on developing without genome (or transcript profile) sequence information, although there is certain practicality, the randomness of mark is strong, poor stability, thus have impact on the accuracy of experimental result.
EST-SSR is a kind of SSR molecular marker based on est sequence or cDNA data mining.Because it is from expressing gene, thus except the institute possessing traditional genomic source SSR marker has superiority, the advantage such as also there is information content high (reflection expressing gene information), development cost are low, versatility is good, thus enhance this and be marked at application in genetic research.At present, this is marked at the research fields such as analysis of genetic diversity, cultivar identification, genetic map construction, gene (QTL) location and obtains good application.At present, for improving the detection efficiency of SSR technology in the crops such as corn, wheat, millet, having carried out the correlative study of core primers group, having achieved good effect.Core primers group refers to a set of primer that uniform fold full-length genome, polymorphism are high, stability is strong, reproducible, utilizes that it carries out analysis of genetic diversity, cultivar identification has quick, easy, that accuracy is high advantage.At present, sponge gourd there is no the specific molecular marker based on genome or the exploitation of transcript profile sequence, and therefore, exploitation sponge gourd EST-SSR core primers group, has become problem in the urgent need to address in sponge gourd breeding, production and kind protection field.
Summary of the invention
One object of the present invention is to provide a set of EST-SSR core primers group based on the exploitation of sponge gourd transcript profile sequence.
Another object of the present invention is to provide the above-mentioned application of EST-SSR core primers group in the analysis of sponge gourd Genetic Diversity of Germplasm, variety of luffa Genetic lineages or variety of luffa qualification based on the exploitation of sponge gourd transcript profile sequence.
The technical solution used in the present invention is:
Based on the EST-SSR core primers group of sponge gourd transcript profile sequence exploitation, it comprises 50 pairs of primers, and its nucleotide sequence is as shown in sequence table SEQ ID NO.1 ~ 100.
Utilize above-mentioned based on sponge gourd transcript profile sequence exploitation EST-SSR core primers group carry out the analysis of sponge gourd Genetic Diversity of Germplasm, variety of luffa Genetic lineages or variety of luffa qualification method, comprise the steps:
(1) sponge gourd sample gene group DNA to be measured is extracted;
(2) the testing sample DNA extracted with step (1), for template, utilizes primer shown in sequence table SEQ ID NO.1 ~ 100 to carry out pcr amplification, obtains pcr amplification product;
(3) pcr amplification product of step (2) is adopted employing 6% modacrylic phthalein amine detected through gel electrophoresis, the colour developing of silver dye, statistic mixed-state result;
(4) statistics of step (3) is utilized to carry out the analysis of sponge gourd Genetic Diversity of Germplasm, variety of luffa Genetic lineages or variety of luffa qualification.
Described pcr amplification reaction system (20 μ L) comprising: 2 μ L10 × buffer; 0.5 μ L Taq enzyme (2U μ L
-1); 0.4 μ L dNTP (10mmolL
-1); Each 0.1 μ L (the 50pmoL μ L of upstream and downstream primer
-1); 1 μ L template DNA (50ng μ L
-1); 15.9 μ L ddH2O.PCR program is Touch-down PCR:94 DEG C denaturation 3min; 94 DEG C of 30S, 65 DEG C of (-1 DEG C/cycle) 1min, 72 DEG C of 1min, 15 circulations; 94 DEG C of 30S, 50 DEG C of 1min, 72 DEG C of 1min, 25 circulations; 72 DEG C of ends extend 10min.
The analysis of described sponge gourd Genetic Diversity of Germplasm or variety of luffa Genetic lineages analytical procedure are: use NTSYS-pc2.l0e software carries out the cluster analysis based on UPGMA method; Use the number of alleles (Na) of each primer of POPGENE computed in software, gene diversity (h) and Shannon value (I).
Described variety of luffa authentication method is: add up the amplification situation of each kind in primer, constructed dna finger printing, one by one more each primer sites difference, differential primer number >=2, judges that two kinds are as different varieties; Differential primer number=1, judges that two kinds are as approximate kind; Differential primer number=0, judges that two kinds are as same breed.
The invention has the beneficial effects as follows: 50 pairs of EST-SSR core primers of the present invention have rich polymorphism, amplification stable, reproducible, be convenient to the advantages such as statistics, simultaneously because these primers are all from sponge gourd expressing gene, the variation of functional sequence in genome can be reflected, thus better can distinguish the diversity of sponge gourd genetic germplasm.This core primers group can be used for the fields such as variety of luffa qualification, the analysis of kind Genetic lineages and Genetic Diversity of Germplasm analysis; be conducive to the legitimate rights and interests protecting breeder, producers and consumers, promote the sound development that sponge gourd genetic breeding and sponge gourd are produced.
Accompanying drawing explanation
Fig. 1 is to the cluster analysis of 46 parts of sponge gourd materials according to 50 pairs of sponge gourd EST-SSR core primers groups.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated, but be not limited thereto.
Embodiment
One, the exploitation of sponge gourd EST-SSR core primers group
1, material
The evaluation of 46 parts of core authors materials (table 1) for sponge gourd EST-SSR core primers group and the qualification of resource genetic diversity is filtered out from the sponge gourd germ plasm resource that our unit preserves.46 parts of sponge gourd resource materials comprise 36 parts of angular sponge gourd, 10 parts of luffa-smooth loofahs; Wherein 25 parts of materials are from China, and 8 parts of materials are from Thailand, and 7 parts of materials, from Malaysia, separately have 6 parts of material the unknown sources.
For sponge gourd resource analysis of genetic diversity material information in table 1 the present invention
Note :-represent unknown source.
2, the extraction of sponge gourd genomic dna
Adopt the simple and easy SDS method of improvement to extract sponge gourd leaves genomic DNA, step is as follows:
(1) proper amount of fresh blade is got, put into 2ml centrifuge tube, add liquid nitrogen, with micro-grinding rod or Philips-type screwdriver, blade is stampped powdered, add 600 microlitre 1.5%SDS trace extract (0.5%SDS, 250mmol/L NaCL, 25mmol/LEDTA, 200mmol/L Tris-HCL damping fluid, pH7.5);
(2) sample hose is placed in 65 DEG C of water-baths and hatches 30 minutes, put upside down mixing once every 5-10 minute;
(3) take out sample, be cooled to room temperature, add the chloroform/primary isoamyl alcohol/ethanol (76:4:20) of equal-volume (600-700 microlitre), vibrate 10 minutes, fully mix;
(4) by centrifugal for sample hose 12000rpm 12 minutes, carefully supernatant liquor is transferred in another 1.5ml centrifuge tube;
(5) add equal-volume Virahol or two volumes dehydrated alcohol precipitation, leave standstill centrifugal 5 minutes of 3-5 minute, 12000rpm;
(6) remove supernatant, precipitate with the ethanol rinse of 0.5ml70%, air-dry, be dissolved in 50-100 μ l TE.
3, pcr amplification
The reaction system (20 μ L) of PCR comprises: 2 μ L10 × buffer, 0.5 μ L Taq enzyme (2U μ L
-1), 0.4 μ L dNTP (10mmolL
-1), each 0.1 μ L (the 50pmoL μ L of upstream and downstream primer
-1), 1 μ L template DNA (50ng μ L
-1), 15.9 μ L ddH2O.
PCR program is Touch-down PCR:94 DEG C denaturation 3min; 94 DEG C of 30S, 65 DEG C of (-1 DEG C/cycle) 1min, 72 DEG C of 1min, 15 circulations; 94 DEG C of 30S, 50 DEG C of 1min, 72 DEG C of 1min, 25 circulations; 72 DEG C of ends extend 10min.
4, polyacrylamide gel electrophoresis detects
(1) configuration of reagent
1 × 6% acrylamide stock solution (100mL): 5.7g acrylamide, 0.3g Bis, 12g urea, 10mL10 × TBE, adds ddH
2o is settled to 100mL; Before encapsulating, every 10mL glue, adds 10% ammonium persulphate 70 μ L, TEMED3 μ L.
Staining fluid (1 ×): 0.1%AgNO
3
Developing solution (1 ×): 0.5g NaOH, 0.019g sodium tetraborate, 0.4mL formaldehyde (with front adding), ddH
2o is settled to 100ml.
(2) preparation of polyacrylamide gel
First sheet glass bottom end opening sepharose is sealed, treat that it solidifies, 1 × 6% acrylamide soln prepared is stirred evenly, sheet glass is become the angle of about 30 ° with desktop, slowly injecting glue, avoid the generation of bubble.Setting level sheet glass after filling makes it become the angle of about 10 ° with desktop, inserts suitable comb.After treating that it solidifies 0.5h, carefully extract comb.Repeatedly rinse loading wells with distilled water, remove the unnecessary material do not solidified.
(3) electrophoresis
Sheet glass is fixed on Vertial electrophorestic tank, adds appropriate 1 × tbe buffer liquid.After PCR terminates, take out sample, often pipe adds the loading buffer that 3 μ L contain bromjophenol blue and the blue or green two kinds of indicator of dimethylbenzene.3 μ L sample loadings are taken out with microsyringe.
(4) argentation dyeing
After electrophoresis, sheet glass is pulled down from electrophoresis chamber, take out gel, rinsing twice in distilled water, at every turn about 1min.Then proceed in staining fluid, jog on shaking table, dyeing 10min.After dyeing terminates, gel is transferred to rinsing twice in distilled water, at every turn about 1min.Then proceed in developing solution and develop the color, proceed in clear water after painted and preserve, gel imaging, record result.
5, EST-SSR primer development and screening
Utilize MIcroSAtellite identification tool (MISA,
http:// pgrc.ipk-gatersleben.de/misa/)software scans SSR site from the sponge gourd Unigene spliced, and screening criteria is that 2 bases at least repeat six times, and 3 bases at least repeat five times, 4,5, and 6 bases at least repeat four times and are just identified as SSR site.Obtain SSR site 12 altogether, 932, utilize Primer premier6.0 (PREMIER Biosoft International, Palo Alto, CA) to carry out design of primers to these candidate locus, successful design 8,523 pairs of primers.Synthesis 641 primers wherein, utilize angular sponge gourd material S1174, and the F1 hybrid of luffa-smooth loofah material 93075 and the two hybridization detects the polymorphism of primer.
6, data analysis
Use NTSYS-pc2.l0e software carries out the cluster analysis based on UPGMA method, uses the number of alleles (Na) of each primer of POPGENE computed in software, gene diversity (h) and Shannon value (I).
7, the determination of core EST-SSR primer sets
Choosing wherein 641 to be synthesized mark, and screening verification is carried out in the F1 of angular sponge gourd kind S1174 and luffa-smooth loofah kind 93075 and the two hybridization, result shows, 494 to being marked with amplified production, 201 pairs are marked at show polymorphism between two parents, wherein codominant be marked with 126 right.According to the genetic linkage maps that we build, from 126 to codominant primer pair, filter out 50 and the primer of each linkage group is stablized, is uniformly distributed to amplification as core EST-SSR primer sets, in table 2.
Table 2 sponge gourd 50 pairs of core EST-SSR primer sets
8, the validity check of sponge gourd EST-SSR core primers group
50 pairs of core EST-SSR primer sets are utilized to carry out diversity analysis to from 46 parts of sponge gourd Germplasms (see table 1) of collecting both at home and abroad.Cluster analysis result is shown in Fig. 1, and result shows: it is a class that 36 parts of angular sponge gourd gather, and it is a class that 10 parts of luffa-smooth loofahs gather.Wherein, 36 parts of angular sponge gourd have the trend of carrying out cluster according to source place, two green grass or young crops, fill with village sponge gourd, in clear sponge gourd, S1174, S095, S155,113005, ten thousand precious sponge gourds, full green sponge gourd, S177, medium green sponge gourd, Heshan pork sponge gourd, 113012,113024,113025 is gathered is a large class, these materials are all economized from Chinese Guangdong; In another large class except the husky skin in refined hilllock sponge gourd and elegant field from Chinese Guangdong outside the province, remaining I-01, BridgeGourd, M18, V-1052, V-1052, BlackHank, Abacus, Huaykeaw, Luffa833, I-23, RidgeGourd be Malaysia of sub-country and Thailand all southeast; Except Grand Prize and Wister Chaichan is from except Malaysia and Thailand in other groups, all the other materials 113003,113008, Lianzhou City's perfume sponge gourd, Meizhou pork sponge gourd, magnificent crown wire melon S087,113073 is all from Chinese Guangdong.Materials A 1206 from MinQty, LEELA of Thailand and unknown source in 10 parts of luffa-smooth loofahs is distant with other 7 parts of materials, and except S15 is from except Thailand in other 7 parts of materials, other materials is all from China.Angular sponge gourd and luffa-smooth loofah inter-species heritable difference are very large, and inbred genetic difference is very little.More existing research reports show, angular sponge gourd and luffa-smooth loofah genetic variation narrow, but utilize 50 pairs of core EST-SSR primer sets in the present invention 46 parts of resource materials can be done well distinguish, this illustrates that core primers group of the present invention can be used for variety of luffa qualification, the analysis of variety of luffa Genetic lineages and the analysis of sponge gourd Genetic Diversity of Germplasm.
<110> Vegetables Inst., Guangdong Academy of Agricultural Sciences
The EST-SSR core primers group that <120> develops based on sponge gourd transcript profile sequence and application
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<210> 65
<211> 20
<212> DNA
<213> artificial sequence
<400> 65
ctctccctgt ttccttctcc 20
<210> 66
<211> 20
<212> DNA
<213> artificial sequence
<400> 66
tttggaacct atgtgcctcc 20
<210> 67
<211> 20
<212> DNA
<213> artificial sequence
<400> 67
ttgtcctctt gcttcccatc 20
<210> 68
<211> 20
<212> DNA
<213> artificial sequence
<400> 68
agctccccaa atggagtttg 20
<210> 69
<211> 20
<212> DNA
<213> artificial sequence
<400> 69
tttgcctcca ccatatgacg 20
<210> 70
<211> 20
<212> DNA
<213> artificial sequence
<400> 70
gcctctttgc caagtgctac 20
<210> 71
<211> 20
<212> DNA
<213> artificial sequence
<400> 71
tccatggccc atggttcatc 20
<210> 72
<211> 19
<212> DNA
<213> artificial sequence
<400> 72
accaggctgg aaaaagggc 19
<210> 73
<211> 22
<212> DNA
<213> artificial sequence
<400> 73
ccaattcacc tccgatccaa tc 22
<210> 74
<211> 20
<212> DNA
<213> artificial sequence
<400> 74
acgttggtta ggcactcgtg 20
<210> 75
<211> 20
<212> DNA
<213> artificial sequence
<400> 75
tgccagatcg aaaccttgtg 20
<210> 76
<211> 20
<212> DNA
<213> artificial sequence
<400> 76
acaattctgt ccacgggaag 20
<210> 77
<211> 20
<212> DNA
<213> artificial sequence
<400> 77
atccaccaac ttccaagcac 20
<210> 78
<211> 20
<212> DNA
<213> artificial sequence
<400> 78
cgaaagtgaa gagaaacccg 20
<210> 79
<211> 22
<212> DNA
<213> artificial sequence
<400> 79
cccatttgct ggtttcagat cg 22
<210> 80
<211> 22
<212> DNA
<213> artificial sequence
<400> 80
cgaaaggcag aatgggaatt cg 22
<210> 81
<211> 20
<212> DNA
<213> artificial sequence
<400> 81
tcctcctcct ttttcccttc 20
<210> 82
<211> 20
<212> DNA
<213> artificial sequence
<400> 82
tctgagacga gaatgccaac 20
<210> 83
<211> 20
<212> DNA
<213> artificial sequence
<400> 83
ttccgatcac tgtcagcttg 20
<210> 84
<211> 20
<212> DNA
<213> artificial sequence
<400> 84
tccacatccc ctttttccag 20
<210> 85
<211> 20
<212> DNA
<213> artificial sequence
<400> 85
atttactcac tctgcttcgc 20
<210> 86
<211> 22
<212> DNA
<213> artificial sequence
<400> 86
aggcaaattc taccaaaaac ac 22
<210> 87
<211> 20
<212> DNA
<213> artificial sequence
<400> 87
aactacttga agccaggccc 20
<210> 88
<211> 20
<212> DNA
<213> artificial sequence
<400> 88
tgatctggca ctagctgcac 20
<210> 89
<211> 20
<212> DNA
<213> artificial sequence
<400> 89
ttcttctcct caggcacagg 20
<210> 90
<211> 21
<212> DNA
<213> artificial sequence
<400> 90
ctgattcgga ggaattcgag g 21
<210> 91
<211> 20
<212> DNA
<213> artificial sequence
<400> 91
cccgtaactc ggttaattcg 20
<210> 92
<211> 20
<212> DNA
<213> artificial sequence
<400> 92
tctgatccgt cattgccatc 20
<210> 93
<211> 20
<212> DNA
<213> artificial sequence
<400> 93
ggtggacctg caagattatg 20
<210> 94
<211> 22
<212> DNA
<213> artificial sequence
<400> 94
attcttccat ctcatttgtt gg 22
<210> 95
<211> 20
<212> DNA
<213> artificial sequence
<400> 95
tctacccgga gcctctcttc 20
<210> 96
<211> 20
<212> DNA
<213> artificial sequence
<400> 96
aacatggagg tctggaggag 20
<210> 97
<211> 20
<212> DNA
<213> artificial sequence
<400> 97
cgctgttgga ctgacatacg 20
<210> 98
<211> 20
<212> DNA
<213> artificial sequence
<400> 98
cattggagga gatggaaggc 20
<210> 99
<211> 20
<212> DNA
<213> artificial sequence
<400> 99
cttggatgca aagtgcttgt 20
<210> 100
<211> 20
<212> DNA
<213> artificial sequence
<400> 100
atggaagcac ccattttgag 20
Claims (10)
1., based on the EST-SSR core primers group of sponge gourd transcript profile sequence exploitation, it is characterized in that: described primer sets comprises 50 pairs of primers, and its nucleotide sequence is as shown in sequence table SEQ ID NO.1 ~ 100.
2. the application of EST-SSR core primers group in sponge gourd Genetic Diversity of Germplasm is analyzed based on the exploitation of sponge gourd transcript profile sequence according to claim 1.
3. the application of EST-SSR core primers group in variety of luffa Genetic lineages based on the exploitation of sponge gourd transcript profile sequence according to claim 1.
4. the application of EST-SSR core primers group in variety of luffa qualification based on the exploitation of sponge gourd transcript profile sequence according to claim 1.
5. utilize described in claim 1 based on sponge gourd transcript profile sequence exploitation EST-SSR core primers group carry out the analysis of sponge gourd Genetic Diversity of Germplasm, variety of luffa Genetic lineages or variety of luffa qualification method, comprise the steps:
(1) sponge gourd sample gene group DNA to be measured is extracted;
(2) the testing sample DNA extracted with step (1), for template, utilizes primer shown in sequence table SEQ ID NO.1 ~ 100 to carry out pcr amplification, obtains pcr amplification product;
(3) pcr amplification product of step (2) is adopted detected through gel electrophoresis, statistic mixed-state result;
(4) statistics of step (3) is utilized to carry out the analysis of sponge gourd Genetic Diversity of Germplasm, variety of luffa Genetic lineages or variety of luffa qualification.
6. method according to claim 5, is characterized in that, the analysis of described sponge gourd Genetic Diversity of Germplasm or variety of luffa Genetic lineages analytical procedure are: use NTSYS-pc 2.l0e software carries out the cluster analysis based on UPGMA method; Use the number of alleles (Na) of each primer of POPGENE computed in software, gene diversity (h) and Shannon value (I).
7. method according to claim 5, is characterized in that, described variety of luffa authentication method is: add up the amplification situation of each kind in primer, constructed dna finger printing, more each primer sites difference one by one, differential primer number >=2, judge that two kinds are as different varieties; Differential primer number=1, judges that two kinds are as approximate kind; Differential primer number=0, judges that two kinds are as same breed.
8. method according to claim 5, is characterized in that, step (3) adopts 6% modacrylic phthalein amine detected through gel electrophoresis, the colour developing of silver dye.
9. method according to claim 5, is characterized in that, 20 μ L pcr amplification reaction systems comprise: 2 μ L 10 × buffer; 0.5 μ L concentration is 2U μ L
-1taq enzyme; 0.4 μ L concentration is 10mmol L
-1dNTP; Concentration is 50 pmoL μ L
-1the each 0.1 μ L of upstream and downstream primer; 1 μ L concentration is 50 ng μ L
-1template DNA; 15.9 μ L ddH
2o.
10. method according to claim 5, is characterized in that, pcr amplification program is Touch-down PCR:94 DEG C denaturation 3min; 94 DEG C of 30S, 65 DEG C, each circulation reduction by 1 DEG C, 1min, 72 DEG C of 1min, 15 circulations; 94 DEG C of 30S, 50 DEG C of 1min, 72 DEG C of 1min, 25 circulations; 72 DEG C of ends extend 10min.
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| CN105203670B (en) * | 2015-11-02 | 2017-03-22 | 黑龙江大学 | Establishing method of HPLC (high performance liquid chromatography) fingerprints of loofah sponge |
| CN105755117B (en) * | 2016-03-03 | 2019-02-12 | 广东省农业科学院蔬菜研究所 | The nucleotide primer of Rapid identification angular sponge gourd ' refined green No. 6 ' hybrid seed purity combines and detection method |
| CN105821154B (en) * | 2016-06-03 | 2019-03-08 | 福建省农业科学院作物研究所 | A kind of SSR primer and its method for the identification of sponge gourd hybrid seed purity |
| CN106011251B (en) * | 2016-06-07 | 2020-05-19 | 山西农业大学 | Primer group, kit and method for identifying genetic population structure of hemerocallis plant |
| CN108359740A (en) * | 2018-04-28 | 2018-08-03 | 江苏省中国科学院植物研究所 | EST-SSR molecular labelings and its application are developed based on Lycoris aurea transcript profile sequence |
| CN111304358B (en) * | 2020-04-22 | 2022-09-09 | 福建省农业科学院作物研究所 | EST-SSR primer developed based on wax gourd transcriptome sequence and application thereof |
| CN111961740B (en) * | 2020-06-29 | 2024-03-26 | 湖南省蔬菜研究所 | SSR primer and method for identifying purity of early-optimal luffa hybrid seeds |
| CN113969323B (en) * | 2021-10-14 | 2022-08-02 | 广东省农业科学院蔬菜研究所 | SNP molecular marker related to luffa melon length and application thereof |
| CN114457184A (en) * | 2022-02-21 | 2022-05-10 | 广东省农业科学院蔬菜研究所 | SNP molecular marker and KASP primer for identifying color of luffa seed coat and application thereof |
| CN117385086B (en) * | 2023-11-27 | 2024-04-16 | 广东省农业科学院蔬菜研究所 | SSR (simple sequence repeat) marker primer for melon germplasm resources and construction method of fingerprint |
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