CN103773866A - Molecular marker, primer, kit and detection method for rapidly detecting flue-cured tobacco scent - Google Patents
Molecular marker, primer, kit and detection method for rapidly detecting flue-cured tobacco scent Download PDFInfo
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Abstract
The invention relates to a molecular marker, a primer, a kit and a detection method for rapidly detecting a flue-cured tobacco scent. About 120 pairs of desoxyribonucleic acid segments with basic sizes are obtained by adopting a random amplified polymorphic desoxyribonucleic acid (RAPD) technology. The segments are cloned by using bacterial plasmids M13; a base sequence is determined to be 1236bp by using an automatic nucleic acid sequence analysis instrument. On the basis of the sequence, a pair of oligomeric nucleotide primers is artificially synthesized, and comprises probes capable of detecting the scent style of the flue-cured tobacco material, namely detecting strong-flavor character expression, and detecting a strong-flavor gene for flue-cured tobacco breeding. The method is simple in operation, reliable in structure, and fast in detection speed, and is not affected by the growth and development period and the environment of a plant, and the identification efficiency can be greatly improved.
Description
Technical field
The invention belongs to biological technical field, particularly a kind of molecule marker for rapid detection flue-cured tobacco flavor types, primer, test kit and detection method.
Background technology
Tobacco leaf is the basis of Cigarette Industrial Enterprise, quality of tobacco and odor type be important role in cigarette composition design, and tabacco fragrance is the important factor of weighing raw tobacco material quality and operability, quality in its quality depends on the fragrance of tobacco leaf to a great extent, therefore, the fragrance matter in research tobacco leaf has great importance for raising quality of tobacco, monitoring cigarette product quality.
The Jiang Chengkang chief of State Tobacco Monopoly Bureau in 2010 proposes to pay much attention to improve quality of tobacco in national tobacco working conference, strives for quality of tobacco and specializes; Will be according to the requirement of " make the most of the advantage, exploit potentialities, plug a gap, satisfy the demands ", give full play to cigarette district of China ecotope diversity advantage, scent type tobacco leaf will further show characteristic, Luzhou-flavor tobacco leaf will have new raising, new district exploitation will have distincter odor type feature, promotes each odor type style tobacco leaf coordinated development.
Fragrance is material incentive people's sense of smell, gustatory organ and the sensation of the pleasant that produces, and it is fragrance and jealous comprehensive.And the fragrance of tobacco is the multiple coefficient result of aroma substance with specific fragrance characteristic, different tobacco types and kind or identical kind are under different ecotopes and cultivation condition, composition, content and the ratio of its aroma component are not quite similar, and therefore show different Type of aromas.But kind is the basic factor that tobacco leaf odor type style forms, specific ecological condition and suitable cultivation technique in conjunction with under, the qualitative characteristics of certain species and style just can give full expression to, and just may form certain characteristic.Therefore, scientific worker thinks that the odor type of flue-cured tobacco is mainly subject to the impact of kind, and the division of flue-cured tobacco cultivars (being) odor type, for instructing cigarette composition can play great function.
Summary of the invention
The object of the invention is to overcome the defect existing in existing detection technique, a kind of molecule marker for rapid detection flue-cured tobacco flavor types, primer, test kit and detection method are provided.The present invention will speed up the application of molecular marker assisted selection in odor type style location.
For a molecule marker for rapid detection flue-cured tobacco flavor types, sequence is as SEQ NO:1.
A pcr amplification method primer used for rapid detection flue-cured tobacco flavor types, sequence is as follows:
Forward sequence F:GTAACTCCCGAGACG,
Reverse sequence R:ATTGATTGGTCAAAGA.
A test kit for rapid detection flue-cured tobacco flavor types, comprises above-mentioned primer, and carries out PCR and react required all ingredients.PCR reacts required all ingredients and comprises: 10 × Buffer, dNTP, MgCl
2, TaqDNA polysaccharase.
A pcr amplification method for rapid detection flue-cured tobacco flavor types,
Take above-mentioned nucleic acid molecule as primer, take flue-cured tobacco genomic dna to be detected as template, carry out pcr amplification, amplify the DNA fragmentation of 1236bp size, show that detected flue-cured tobacco has Luzhou-flavor gene and exists and express.
PCR system comprises 10 × Buffer, 0.2mmol/L dNTP, 2.0mmol/L MgCL
2, 2U TaqDNA polysaccharase, 50ng template DNA, random primer 0.5 μ mol/L, random primer used is numbered S101-S200; Water is filled it up with to 25 μ l.
PCR reaction conditions: front 12 circulations are ℃ sex change 30s → 65,94 ℃ of denaturation 2min → 94 ℃ annealing 30s, extend 1min with speed rising → 72 ℃ of 0.7 ℃/each circulation; Rear 23 circulations are that 1min → 72 ℃ extension 5min is extended in ℃ annealing 30s → 72 ℃, 94 ℃ of sex change 30s → 56.
Flue-cured tobacco cultivars to be detected comprises: the large gold dollar of safflower, cloud and mist 85, K326, NC297, NC89, dark green No. one, CB-1, C2, F1-35, NC55, No. 3, Hunan cigarette, Zhongyan-100, cloud and mist 97, No. 7, Henan cigarette, NC89, cloud and mist 87 and Guangdong cigarette 97.
Technical problem to be solved by this invention specifically realizes by following technical scheme:
1, adopt Random amplified polymorphic DNA (RAPD) technology, use random primer 100(S101-S200) the individual pcr amplification that carries out, in primer, S156 can amplification region separates the flue-cured tobacco of Luzhou-flavor and scent type, has obtained the molecule marker S156 with the gene linkage of tobacco odor type
1236, its size is about 1236bp(Fig. 1).
2, after the molecule marker obtaining being cloned with bacterial plasmid M13, measure this mark (S156 with nucleic acid automatic analysis sequence instrument in step 1
1236) base form and sequence, show that this molecule marker is made up of and particular sorted order 1236 pairs of bases;
3, the DNA sequence dna of measuring in step 2 is analyzed, and contrast with random primer S156, be foundation according to obtained sequence, utilize DNA synthesizer, synthetic oligonucleotide (forward sequence F:GTAACTCCCGAGACG, reverse sequence R:ATTGATTGGTCAAAGA), it can detect Luzhou-flavor flue-cured tobacco; And show flue-cured tobacco with the DNA sequence dna of about 1236bp size and had Luzhou-flavor proterties and Luzhou-flavor gene.
4, increase with 17 flue-cured tobacco material genomic dnas of primer pair synthetic in step 3, wherein Luzhou-flavor flue-cured tobacco material has showed the DNA sequence dna of about 1236bp size jointly, and scent type flue-cured tobacco material does not all show this sequence (Fig. 2).
5, compared with conventional art, detection method provided by the present invention, be not only and first utilize the method for molecule marker in genetically engineered to detect flue-cured tobacco flavor types style, and the feature such as the impact that method is easy, reliable, speed soon, is not subject to vine growth and development period and environment detecting, just can identify in early days (having made up the defect that could identify in the ripening stage in traditional method) in seedling stage, greatly improve determination rates.
Accompanying drawing explanation
Fig. 1 is the amplification of random primer S156 to flue-cured tobacco material;
Fig. 2 is the qualification result of primers F/R in flue-cured tobacco material,
From left to right be followed successively by the large gold dollar of safflower, cloud and mist 85, K326, NC297, NC89, dark green No. one, CB-1, C2, F1-35, NC55, No. 3, Hunan cigarette, Zhongyan-100, cloud and mist 97, No. 7, Henan cigarette, NC89, cloud and mist 87 and Guangdong cigarette 97.
Embodiment
Following examples are intended to further illustrate the present invention, and unrestricted the present invention.
Embodiment 1: the process that the inventive method is implemented
(1) preparation of flue-cured tobacco material: in floating seedling tray, in the time that tobacco seedling growth arrives 3-4 sheet true leaf, each material blade is got 10g left and right by the planting seed of 17 flue-cured tobacco materials.
(2) extraction of genomic dna: utilize CTAB method to extract genomic dna, by cryodesiccated tobacco leaf grind into powder, add the CTAB extract of 65 ℃ of preheatings, more than 65 ℃ of insulation 45min, between or after the centrifugal 20min of mix → 12000r/min of jog room temperature, get the isopyknic chloroform of supernatant liquor → add: primary isoamyl alcohol (24:1), more than mixing 15min gently, the centrifugal 20min of 12000r/min room temperature, get supernatant liquor and move in new centrifuge tube → add the Virahol of precooling, shake gently 5min, after the centrifugal 10min of 8000r/min room temperature, remove supernatant → precipitation 75% ethanol and 10mmol/L KAC extracting 2～3 times (each the centrifugal 10min of 8000r/min room temperature) → add 95% ethanol of precooling, turn upside down gently, after the centrifugal 20min of 12000r/min room temperature, abandon ethanol, vacuum is drained or is naturally dried → add 200ulTE buffer, beat gently and make resolution of precipitate → add 1ul10mg/mL Rnase37 ℃ water-bath, insulation 1h, remove RNA → DNA extraction thing be put in-20 ℃ of refrigerators, preserve for subsequent use.
(3) pcr amplification: carry out RAPD-PCR amplification take flue-cured tobacco genomic dna as template, PCR system comprises 10 × Buffer, 0.2mmol/L dNTP, 2.0mmol/L MgCL
2, 2U TaqDNA polysaccharase, 50ng template DNA, random primer 0.5 μ mol/L; Add water to 25 μ l.PCR reaction conditions: front 12 circulations are the speed rising of ℃ sex change 30s → 65,94 ℃ of denaturation 2min → 94 ℃ annealing 30s(with 0.7 ℃/each circulation) → 72 ℃ of extension 1min; Rear 23 circulations are that 1min → 72 ℃ extension 5min is extended in ℃ annealing 30s → 72 ℃, 94 ℃ of sex change 30s → 56; Wherein random primer used is numbered the raw work in S101-S200(Shanghai).
(4) electrophoresis detection and order-checking: the product electrophoresis detection after amplification, amplify the feature band about about 1236bp, by this section of sequence clone, in bacterial plasmid M13, the base of measuring this mark with nucleic acid automatic analysis sequence instrument forms and sequence.
Embodiment 2: the inventive method is implemented the contrast of rear flue-cured tobacco material odor type
1, choose 17 parts of flue-cured tobacco materials, the odor type of these materials is in table 1.
2, the extraction of flue-cured tobacco genomic dna and pcr amplification detect with embodiment 1, and it is foundation according to obtained 1236bp sequence that pcr amplification detects primer used, utilizes DNA synthesizer, synthetic oligonucleotide,
Forward sequence F:GTAACTCCCGAGACG, reverse sequence R:ATTGATTGGTCAAAGA.
3, the inventive method implement after the contrast of flue-cured tobacco material odor type, the results are shown in Table 1 and Fig. 2.
Table 1F/R primer detected result and the contrast of tobacco material odor type
| Swimming lane | Tobacco material | Odor type | F/R primer detected result | Swimming lane | Tobacco material | Odor type | F/R primer detected result |
| 1 | The large gold dollar of safflower | Scent type | Without |
10 | NC55 | Luzhou-flavor | There is |
| 2 | Cloud and mist 85 | Scent type | Without |
11 | No. 3, Hunan cigarette | Luzhou-flavor | There is |
| 3 | k326 | Luzhou-flavor | There is |
12 | Zhongyan-100 | Luzhou-flavor | There is |
| 4 | NC297 | Luzhou-flavor | There is |
13 | Cloud and mist 97 | Scent type | Without |
| 5 | NC89 | Luzhou-flavor | There is |
14 | No. 7, Henan cigarette | Luzhou-flavor | There is |
| 6 | Dark green No. one | Scent type | Without |
15 | NC89 | Luzhou-flavor | There is |
| 7 | CB-1 | Scent type | Without |
16 | Cloud and mist 87 | Scent type | Without |
| 8 | C2 | Scent type | Without |
17 | Guangdong cigarette 97 | Luzhou-flavor | There is |
| 9 | F1-35 | Scent type | Without band | ? | ? | ? | ? |
A?1cccatgacca?atatgagctc?gaagcttcct?tc
gtaactcc?cgagacgagc?cgtttatcat?tacgataggt?gtcaagtgga
B?1cccatgacca?atatgagctc?gaagcttcct?cctcacaatc?tttcattgtt?cgtttatcat?tacgataggt?gtcaagtgga
A81?tgaggcatcc?taacagaccg?gtagacttga?tctatttcat?tatattccat?ccatatccca?attccattca?accttgttcc
B81?tggttaaact?ctactgcggt?ttcaactctt?tctatttcat?tatattccat?ccatatccca?attccattca?aagcgcgtag
A161?aagtaccttt?aaaaccattg?tacatgacct?caaatctttg?catgatcggg?gatcaattcg?ccgttcccaa?gatctcatag
B161?aagtaccttt?aaaaccattg?tacatgacct?caaatctttg?catgatcggg?gatcaattcg?ccgttcccaa?gatctcatag
A241?agttcaattc?ccctctgggg?ggcagaggcc?acaacacgaa?cctccagtca?cgccatctat?cctcacaatc?tttcattgtt
B241?ccgctctacc?ggtacttttt?gccgaccctc?caggagattc?cccaaaaggg?atacatctat?cctcacaatc?tttcattgtt
A321?actagccaat?atgcttttct?ctcatgcctt?tcttcgttca?tggttcgata?ttctggtgtc?ctaggcgtag?aggaaccaca
B321?actagccaat?atgcttttct?ctcatgcctt?tcttcgttca?tggttcgata?ttctggtgtc?ctaggcgtag?aggaaccaca
A401?ccaatccatc?ccgaacttgg?tggttaaact?ctactgcggt?ttcaactctt?tgacaacatg?aaaaaaccaa?aagctctgcc
B401?ccaatccatc?ccgaacttgg?tggttaaact?ctactgcggt?cgtaggtact?gtattgacgatcggaaccaa?aagctctgcc
A481?atgcagtgct?cgccaggatg?agtatatctg?ataaaaagct?agacttctta?taacacctct?attggctagt?aaatgaaaaa
B481?atgccgagct?cataaggatg?agtatatccc?tgaaaagct?agacttctta?taacaccatc?tggatctagt?aaatgaaaaa
A561?gacgatactg?aaaaaaaaaa?taggggaggt?actttttcaa?cctgcggaaa?cattcttatt?tatgaaaacg?aatagctcga
B561?tagcttgcac?gtaagcagct?acgtaaagct?actttttcaa?cctgcggaaa?tgcatagggg?tatgaaaacg?aatagctcga
A641?tatagagata?aacgcgcctt?cacatcttct?taacccgaaa?tggctgggga?gaggaaaggt?tccttttttt?gagggtactc
B641?tatagagata?aacgcgcctt?cacatcttct?taacccgaaa?tggctgggga?gaggaaaggt?tccttttttt?gagggtactc
A721?ccgggaacag?atccagtgga?gacggggtgg?ggcctgtagc?tcccttctct?cccacttcac?acctcggaac?gcacccttct
B721?ccgggaacag?atccagtgga?gacggggtgg?ggcctgtagc?tcccttctct?cccacttcac?acctcggaac?gcgaggttct
A801?gggttcgaat?taacagccga?ccctcctcgc?ccgctctacc?tgaggaacaa?ccacaaccgg?caggagattc?cccaaaaggg
B801?gggaattcat?taacagcact?tcctcctcgc?ccgcaacacc?tgaggaaggttaacaaccgg?caggagattc?cccaaaaggg
A881?atagcgaacc?aaaagctatg?gaacttgggt?gtgggtcttt?tgtcgaaatg?gaatggcttt?tctttttctc?tttttattta
B881?atagcgaacc?aaaagctatg?gaacttgggt?gtgggtcttt?tgtcgaaatg?gaattcataa?tcgactagtg?tcaagattta
A961?agaacagaat?gagatagaat?gaacgggtta?acgggcaaag?agagggaacc?tcaaagtggc?tttaatatcc?ctttggcagc
B961?tgctggatcg?tactagcggt?acgtacgtaa?ccgttcaact?ggggaaatcg?tatattgcaa?tttaatatcc?ctttggcagc
A1041?caggcttggg?cagaatagca?gagcaagtat?tagtagcata?acaaaaaagc?cttcctcgtc?attaatatct?ttgctcgcgg
B1041?caggcttggg?cagaatagca?gagcaagtat?tagtagcata?acaaaaaagc?cttcctcgtc?attaatatct?ttgctcgcgg
A1121?caattgtgac?cgagcccctt?acggtgtcgg?ctctcgggag?aatcgatgac?agaatcaatc?tgca
tctttg?accaatcggt
B1121?caattgtgac?cgagcccctt?acggtgtcgg?ctctcgggag?aatcgatgac?agaatcaatc?tcaaagtggccaaagagagg
A1201?tgtaaatagc?cccagggcta?tggaacaaag?gattat
B1201?tgtaaatagc?cccagggcta?gagatagaat?gaacgggtta?acggggaacc
Be more than the 1236 pairs of bases amplifying form with known Luzhou-flavor related gene base sequence to compare (underscore is primer location) A be this test extension increasing sequence, B is that known Luzhou-flavor related gene base sequence (draws from periodical Mol.Gen.Genet.180,1-4,1980); Shade represents the different piece of both sequences.
Claims (8)
1. for a molecule marker for rapid detection flue-cured tobacco flavor types, it is characterized in that, sequence is as SEQ NO:1.
2. a pcr amplification method primer used for rapid detection flue-cured tobacco flavor types, is characterized in that, sequence is as follows:
Forward sequence F:GTAACTCCCGAGACG,
Reverse sequence R:ATTGATTGGTCAAAGA.
3. a test kit for rapid detection flue-cured tobacco flavor types, is characterized in that, comprises primer claimed in claim 2, and carries out PCR and react required all ingredients.
4. the test kit of rapid detection flue-cured tobacco flavor types according to claim 3, is characterized in that, PCR reacts required all ingredients and comprises: 10 × Buffer, dNTP, MgCl
2, TaqDNA polysaccharase.
5. a pcr amplification method for rapid detection flue-cured tobacco flavor types, is characterized in that,
Take nucleic acid molecule claimed in claim 2 as primer, take flue-cured tobacco genomic dna to be detected as template, carry out pcr amplification, amplify the DNA fragmentation of 1236bp size, show that detected flue-cured tobacco has Luzhou-flavor gene and exists and express.
6. the pcr amplification method of rapid detection flue-cured tobacco flavor types according to claim 5, is characterized in that,
PCR system comprises 10 × Buffer, 0.2mmol/L dNTP, 2.0mmol/L MgCL
2, 2U TaqDNA polysaccharase, 50ng template DNA, random primer 0.5 μ mol/L, random primer used is numbered S101-S200; Water is filled it up with to 25 μ l.
7. the pcr amplification method of rapid detection flue-cured tobacco flavor types according to claim 5, is characterized in that,
PCR reaction conditions: front 12 circulations are ℃ sex change 30s → 65,94 ℃ of denaturation 2min → 94 ℃ annealing 30s, extend 1min with speed rising → 72 ℃ of 0.7 ℃/each circulation; Rear 23 circulations are that 1min → 72 ℃ extension 5min is extended in ℃ annealing 30s → 72 ℃, 94 ℃ of sex change 30s → 56.
8. the pcr amplification method of rapid detection flue-cured tobacco flavor types according to claim 5, it is characterized in that, flue-cured tobacco cultivars to be detected comprises: the large gold dollar of safflower, cloud and mist 85, K326, NC297, NC89, dark green No. one, CB-1, C2, F1-35, NC55, No. 3, Hunan cigarette, Zhongyan-100, cloud and mist 97, No. 7, Henan cigarette, NC89, cloud and mist 87 and Guangdong cigarette 97.
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| CN107354202A (en) * | 2017-07-10 | 2017-11-17 | 中国烟草总公司郑州烟草研究院 | Primer for identifying flue-cured tobacco K326 combines and kit, application and authentication method |
| CN107354201A (en) * | 2017-07-10 | 2017-11-17 | 中国烟草总公司郑州烟草研究院 | Primer for identifying flue-cured tobacco cloud and mist 97 combines and kit, application and detection method |
| CN107354200A (en) * | 2017-07-10 | 2017-11-17 | 中国烟草总公司郑州烟草研究院 | Primer for identifying dark green No. 1 of flue-cured tobacco combines and kit, application and authentication method |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892304A (en) * | 2010-04-07 | 2010-11-24 | 云南省烟草农业科学研究院 | Method for detecting N gene-controlled tobacco TMV resistance by molecular markers |
-
2014
- 2014-01-15 CN CN201410017753.3A patent/CN103773866B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892304A (en) * | 2010-04-07 | 2010-11-24 | 云南省烟草农业科学研究院 | Method for detecting N gene-controlled tobacco TMV resistance by molecular markers |
Non-Patent Citations (1)
| Title |
|---|
| 任民: "《烟草（N.tabacumL）高香气特征性状的鉴定及其分子标记研究》", 《中国博士学位论文全文数据库（电子期刊）》 * |
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