CN108753796A - A kind of the DB genes and its coding albumen and application of the control limited fruit branch character of cotton - Google Patents
A kind of the DB genes and its coding albumen and application of the control limited fruit branch character of cotton Download PDFInfo
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- CN108753796A CN108753796A CN201810705953.6A CN201810705953A CN108753796A CN 108753796 A CN108753796 A CN 108753796A CN 201810705953 A CN201810705953 A CN 201810705953A CN 108753796 A CN108753796 A CN 108753796A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8262—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield involving plant development
- C12N15/827—Flower development or morphology, e.g. flowering promoting factor [FPF]
Abstract
The present invention relates to a kind of DB genes of control limited fruit branch character of cotton and its coding albumen and applications, belong to cotton gene technical field.The limited fruit branch DB genes of cotton are compared with unlimited fruit branch DB genes, and the 313rd nucleotide sports thymidine A by cytimidine G, and corresponding amino acid sequence is asparagine by Aspartic acid mutations at the 73rd.It is sequenced after carrying out map based cloning on the basis of DB genes provided by the invention are the hybrid Populations in the unlimited fruit branch Type Cotton of wild type and limited fruit branch Type Cotton, obtains the functional gene for controlling limited fruit branch character.In the limited fruit branch DB channel genes cottons of cotton provided by the invention, the cotton variety for obtaining limited fruit branch type and blooming in advance is cultivated, is cultivated for new cotton variety and Cotton Production provides new developing direction.
Description
Technical field
The invention belongs to cotton gene technical fields, and in particular to it is a kind of control the limited fruit branch character of cotton DB genes and
It encodes albumen and application.
Background technology
Cotton is important sources of most important natural fiber crop and albumen, grease in the world.Exactly because cotton
Fiber and seed are closely related with human lives, become one of industrial crops important in the world.The global cotton output value more than 500 hundred million
Dollar, about 4.9 hundred million mu of cultivated area.Five state such as China, India, U.S. accounts for 70% of total yield or more, the advanced plant such as Sino-U.S., Australia
Cotton country, realizes entire mechanization, international competitiveness is strong, leads cotton developing direction.China is maximum cotton in the world
Producing country, country of consumption and textile exporting country.Cotton is not only the Important Economic source of nearly 200,000,000 cotton grower in China, and is related to
The problem of employment of 19000000 weavings and relevant industries practitioner are related to notional economic stability and cotton industry development.With
The continuous of China's human cost is risen, and cotton planting benefit is gradually reduced, and the planting patterns for changing cotton can be utmostly
Raising cotton planting benefit.Due to the characteristic of cotton indeterminate growth, growth period is longer, and the different cotton boll maturity periods are inconsistent, lead
It has caused the recruitment of Cotton Production more and has been difficult to entire mechanization, change the characteristic of cotton indeterminate growth and shortened the fertility of cotton
Phase will can change the planting patterns of cotton, promote entire mechanization process, improve and plant cotton benefit.
Cotton plant type structure and prematureness have larger impact to yield.The most of upland cotton product applied in production at present
Kind is unlimited fruit branch type, and fruit branch includes that multiple fruits are saved, can be with indeterminate growth；The limited fruit branch mutant of cotton is generally also called zero
Formula fruit branch, zero formula fruit branch mutant usually show bud directly differentiation at stem axil in sea island cotton, and in upland cotton
The fruit branch of zero formula fruit branch material is usually a section determinate growth, and top is terminated with 2~3 flowers, with precocious, plant type is compact, flower
The characteristics of phase concentrates.Therefore, the mutation of zero formula fruit branch regulates and controls plant type structure, keeps cotton plant type compact, more to research fruit branch development
It adopts etc. suitable for high-density planting and suitable for machine and has great theoretical and practical significance.But there is no any passes in the prior art
Therefore manual control cotton plant type is also cannot achieve at present in the report of control cotton plant type structure and the gene of Early mature apricot
Yield traits are improved with artificial.
Invention content
In view of this, the purpose of the present invention is to provide a kind of the DB genes and its coding of the control limited fruit branch character of cotton
Albumen and application, the effective control cotton plant type structure of the limited fruit branch DB genes and Early mature apricot, are trained for new cotton variety
It educates and provides new developing direction with Cotton Production.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of limited fruit branch DB genes of cotton, the nucleotide such as sequence table SEQ ID of the gene
Shown in No.1.
The present invention provides a kind of coded sequence of the limited fruit branch DB genes of cotton, the nucleotide of the coded sequence
Sequence is as shown in sequence table SEQ ID No.2.
The present invention provides a kind of coding albumen of the limited fruit branch of cotton, translate to obtain by the coded sequence, the volume
The amino acid sequence of code albumen is as shown in sequence table SEQ ID No.3.
The present invention provides the DB genes, the coded sequence or the coding albumen in the class for regulating and controlling cotton fruit branch
Application in type.
The present invention provides the DB genes, the coded sequence, the recombinant vector, the recombinant bacterium, described turn
The application of gene cell system or the coding albumen in the cotton variety for cultivating limited fruit branch type.
The present invention provides the DB genes, the coded sequence or the coding albumen to cultivate the cotton that blooms ahead of time
Application in kind.
The present invention provides a kind of limited fruit branch DB genes of cotton, the nucleotide such as sequence table SEQ ID of the gene
Shown in No.1.The limited fruit branch DB genes of cotton provided by the invention compared with the DB genes of the unlimited fruit branch Type Cotton of wild type,
There are the 313rd coding mutations, sport thymidine A by cytimidine G, corresponding amino acid sequence is at the 73rd by asparagus fern
Histidine mutations are asparagine, and the DB genes have the biological function for controlling limited fruit branch character.The limited fruit of cotton
Branch DB gene order length is 1013bp, and the DB genes include 4 exons and are spaced in 3 intrones of 4 exons,
The mutation is happened on the 2nd exon.In the limited fruit branch DB channel genes cottons of cotton provided by the invention, render transgenic
The phenotype of cotton becomes limited fruit branch type from unlimited fruit branch type.
Meanwhile the limited fruit branch DB genes of cotton provided by the invention can also effectively improve cotton flowering time, it is demonstrated experimentally that
The more unlimited fruit branch cotton of transgenosis containing the DB genes compares, 7~10d of advance flowering period.As it can be seen that cotton provided by the invention
Spending limited fruit branch DB gene pairs to shift to an earlier date cotton flowering time has biological action.Due to shifting to an earlier date cotton flowering time to later stage cotton boll
Maturation has very great help, and then makes cotton prematurity, shortens the Cotton Production period, is provided newly for Cotton Production with new kind is cultivated
Direction.
Description of the drawings
Fig. 1 is the exon schematic diagram of the limited fruit branch DB genes of cotton provided by the invention；
Fig. 2 is the coded protein sequence schematic diagram of GhNB genes；
Fig. 3 is the electrophoretogram that effect is marked in embodiment 1.
Specific implementation mode
The present invention provides a kind of DB genes of the control limited fruit branch character of cotton, the nucleotide such as sequence tables of the gene
Shown in SEQ IDNo.1.
In the present invention, the limited fruit branch DB gene order length is 1013bp, and the DB genes include 4 exons
With 3 intrones for being spaced in 4 exons, compared with the DB genes of the unlimited fruit branch Type Cotton of wild type, there are the 313rd
Coding mutation sports A by G, and the mutation is happened on the 1st exon (see Fig. 1).
In the present invention, the limited fruit branch DB genes and arabidopsis TFL1/ATC genes are homologous genes, are belonged to
PEBP gene families.
In the present invention, the amplification method of the limited fruit branch DB genes is carried out using regular-PCR method, obtains limited fruit branch
DB genetic fragments.Amplimer for the limited fruit branch DB genes includes forward primer and reverse primer.The forward direction is drawn
The nucleotide sequence of object is as shown in sequence table SEQ ID No.4.The nucleotide sequence of the forward primer such as sequence table SEQ ID
Shown in No.5.The amplification of the limited fruit branch DB genes is to be with the genomic DNA of limited fruit branch island cotton material (new sea 18)
Template amplification.The program of the amplification is as follows：95℃30s；95 DEG C of 5s, 60 DEG C of 30~34s；72℃5min.
The present invention provides a kind of coded sequence (CDS) of the limited fruit branch DB genes of cotton, the coded sequence
Nucleotide sequence is as shown in sequence table SEQ ID No.2.The overall length of the CDS is 525bp.The amplification method of the CDS is preferred
Use the above-mentioned amplimer for the limited fruit branch DB genes to the cDNA in limited fruit branch island cotton material (new sea 18)
For template amplification.The amplification program is the same as DB gene magnifications.
The present invention provides a kind of coding albumen of the limited fruit branch of cotton, translate to obtain by the coded sequence, the volume
The amino acid sequence of code albumen is as shown in sequence table SEQ ID No.3.The coding albumen includes 174 amino acid.
In the present invention, the method that the preparation method of the coding albumen preferably uses recombinant protein expression and purity
It obtains.Recombinant vector prepared by said program is preferably imported in prokaryotes and is induced by the method for the recombinant protein induced expression
Expression.The present invention is not particularly limited the method for the induced expression, using the scheme of induced expression known in the art
?.After induced expression, the present invention will obtain expression liquid and purify.The scheme of the purifying is using known in the art pure
Change scheme.
The present invention provides the DB genes, the coded sequence or the coding albumen to cultivate limited fruit branch type
Cotton variety in application.In the present invention, limited fruit branch type is cultivated using the DB genes, the coded sequence
The method of cotton variety, it is preferred to use conventional hybridization method.
The present invention provides the DB genes, the coded sequence or the coding albumen to cultivate the cotton that blooms ahead of time
Application in kind.The method of the cultivation is same as above, and this will not be repeated here.
With reference to embodiment to a kind of the DB genes and its coding of the control limited fruit branch character of cotton provided by the invention
Albumen and application are described in detail, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
Extract new extra large 18 genomic DNA of the limited fruit branch material of sea island cotton, the amplification method of the limited fruit branch DB genes
It is carried out using regular-PCR method, obtains limited fruit branch DB genetic fragments.Amplimer for the limited fruit branch DB genes includes
Forward primer and reverse primer；F：ATGGCAAAACTGTCAGATCC(SEQ ID No.4)；R：
The amplification of TTAGCGTCTTCTAGCAGCTGT (SEQ ID No.5) described limited fruit branch DB genes is with the limited fruit branch of sea island cotton
New extra large 18 genomic DNA of material is template amplification.The program of the amplification is as follows：95℃30s；95 DEG C of 5s, 60 DEG C 30~
34s；72℃5min.
The limited fruit branch material of sea island cotton new extra large 18 is hybridized with Upland Cotton nakamise 100, F1 is returned with nakamise 43,
BC1F1 generation using DB gene functions label carry out molecular marker assisted selection, DB gene functions labeled primer its be nucleosides
Acid sequence is as follows：
Forward primer 5 '-CAGATGCCACATTTGGTAAG-3 ' (SEQ ID No.6)
Reverse primer 5 '-CTGTGGATCCCTATGTTAGA-3 ' (SEQ ID No.7).
Selected single plant continues to be returned for 4 generations with nakamise 100, is returned and is each selected from generation to generation using DB gene functions label
It selects, it is ensured that DB genes are not lost, while genetic background is returned to nakamise 100, are selfed for selected single plant in BC5, obtain
The BC5F2 single plants of DB gene pures are cultivated in the limited fruit branch upland cotton new lines DB- of the gene containing DB 100, fiber quality
Equal economical characters do not have difference with nakamise 43, but florescence, maturity period do sth. in advance the limited fruit branch types of 7~10d.
Amplification is shown in Fig. 3.From the figure 3, it may be seen that in BC1F1 generations, obtain gene containing DB through PAGE gel electrophoresis.
As can be seen that 100 plant blossom phases in the new lines DB- containing DB genes of culture, maturity period are done sth. in advance, fruit branch class
Type is limited.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Sequence table
<110>The Chinese Academy of Agriculture Science and Technologys Cotton Research Institute
<120>A kind of the DB genes and its coding albumen and application of the control limited fruit branch character of cotton
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1013
<212> DNA
<213> Gossypium hirsutum Linn
<400> 1
atggcaaaac tgtcagatcc tcttgtgttg gggagagtga ttggggatgt tattgatgcc 60
ctctccccat ctgtgaaaat gtcagtcact ttcaacacca acaagcaggt atataatggc 120
catgaatttt ttccatctgc agttactaac aagcctaagg ttgaggttca tggaggtgat 180
atgagatcct ttttcaccct ggtaactata catgtcgcta atactctctt ttcttttctt 240
tttttctgga tttttcaaga gaaaatgtct atcagcaatc atatttttat attgcaggtg 300
atgacagacc cagatgttcc tggtcctagt gacccttacc tgagggagca cttacactgg 360
tacttctctt aatcaaacta catgtataaa aatacaaaat atattaacca ttaatactaa 420
tatataactt ccatagggaa gctgacaatg atctatctat ctatctatat atatataatg 480
actgttaggg ctcataattt ccttggcttg ataaagctaa aaaaaaagga aataagattt 540
catcaaaaca agaagagcac aaatttaagc aatctactga atatttgtac ttgaattgac 600
aatctcacag gatagtgaca gatatccccg gcacaacaga tgccacattt ggtaagtatc 660
ctcttcattc ttcatagaga gagagagaga gagagagaga gagagacaga gagagagaga 720
gagcattata tgatagaaaa tggactaact ttgatggtta aaaatataat tataaaatat 780
atatacacac aggaagggaa atggtgaact acgaaatgcc aaggtcaaac atagggatcc 840
acaggtttgt gttcctcctc ttcaagcaaa aaggcaggca aacagtgaga agcataccgt 900
catcaaggga tcgtttcgat accaggaagt ttgcagaaga aaacgaacta ggggttcctg 960
ttgcagctgt ctatttcaat gctcaaaggg aaacagctgc tagaagacgc taa 1013
<210> 2
<211> 525
<212> DNA
<213> Gossypium hirsutum Linn
<400> 2
atggcaaaac tgtcagatcc tcttgtgttg gggagagtga ttggggatgt tattgatgcc 60
ctctccccat ctgtgaaaat gtcagtcact ttcaacacca acaagcaggt atataatggc 120
catgaatttt ttccatctgc agttactaac aagcctaagg ttgaggttca tggaggtgat 180
atgagatcct ttttcaccct ggtgatgaca gacccagatg ttcctggtcc tagtgaccct 240
tacctgaggg agcacttaca ctggatagtg acagatatcc ccggcacaac agatgccaca 300
tttggtaagg aaatggtgaa ctacgaaatg ccaaggtcaa acatagggat ccacaggttt 360
gtgttcctcc tcttcaagca aaaaggcagg caaacagtga gaagcatacc gtcatcaagg 420
gatcgtttcg ataccaggaa gtttgcagaa gaaaacgaac taggggttcc tgttgcagct 480
gtctatttca atgctcaaag ggaaacagct gctagaagac gctaa 525
<210> 3
<211> 174
<212> PRT
<213> Gossypium hirsutum Linn
<400> 3
Met Ala Lys Leu Ser Asp Pro Leu Val Leu Gly Arg Val Ile Gly Asp
1 5 10 15
Val Ile Asp Ala Leu Ser Pro Ser Val Lys Met Ser Val Thr Phe Asn
20 25 30
Thr Asn Lys Gln Val Tyr Asn Gly His Glu Phe Phe Pro Ser Ala Val
35 40 45
Thr Asn Lys Pro Lys Val Glu Val His Gly Gly Asp Met Arg Ser Phe
50 55 60
Phe Thr Leu Val Met Thr Asp Pro Asp Val Pro Gly Pro Ser Asp Pro
65 70 75 80
Tyr Leu Arg Glu His Leu His Trp Ile Val Thr Asp Ile Pro Gly Thr
85 90 95
Thr Asp Ala Thr Phe Gly Lys Glu Met Val Asn Tyr Glu Met Pro Arg
100 105 110
Pro Asn Ile Gly Ile His Arg Phe Val Phe Leu Leu Phe Lys Gln Lys
115 120 125
Gly Arg Gln Thr Val Arg Ser Ile Pro Ser Ser Arg Asp Arg Phe Asp
130 135 140
Thr Arg Lys Phe Ala Glu Glu Asn Glu Leu Gly Val Pro Val Ala Ala
145 150 155 160
Val Tyr Phe Asn Ala Gln Arg Glu Thr Ala Ala Arg Arg Arg
165 170
<210> 4
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
atggcaaaac tgtcagatcc 20
<210> 5
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
ttagcgtctt ctagcagctg t 21
<210> 6
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
cagatgccac atttggtaag 20
<210> 7
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
ctgtggatcc ctatgttaga 20
Claims (6)
1. a kind of DB genes of the control limited fruit branch character of cotton, the nucleotide sequence such as sequence table SEQ ID of the gene
Shown in No.1.
2. a kind of coded sequence of the limited fruit branch DB genes of cotton described in claim 1, the nucleotides sequence of the coding albumen
Row are as shown in sequence table SEQ ID No.2.
3. a kind of coding albumen of the limited fruit branch of cotton, which is characterized in that the coded sequence described in claim 2 is translated to obtain,
The amino acid sequence of the coding albumen is as shown in sequence table SEQ ID No.3.
4. encoding albumen described in the coded sequence, claim 3 described in DB genes, claim 2 described in claim 1 regulating and controlling
Application in the limited fruit branch type of cotton.
5. encoding albumen described in the coded sequence, claim 3 described in DB genes, claim 2 described in claim 1 cultivating
Application in the cotton variety of limited fruit branch type.
6. encoding albumen described in the coded sequence, claim 3 described in DB genes, claim 2 described in claim 1 cultivating
The application bloomed ahead of time in cotton variety.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810705953.6A CN108753796A (en) | 2018-07-02 | 2018-07-02 | A kind of the DB genes and its coding albumen and application of the control limited fruit branch character of cotton |
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| CN201810705953.6A CN108753796A (en) | 2018-07-02 | 2018-07-02 | A kind of the DB genes and its coding albumen and application of the control limited fruit branch character of cotton |
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| CN108753796A true CN108753796A (en) | 2018-11-06 |
Family
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| CN201810705953.6A Pending CN108753796A (en) | 2018-07-02 | 2018-07-02 | A kind of the DB genes and its coding albumen and application of the control limited fruit branch character of cotton |
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Cited By (1)
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| CN112063741A (en) * | 2020-09-10 | 2020-12-11 | 中国农业科学院棉花研究所 | Cotton fruit node long and short molecular marker and application thereof |
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| CN104561284A (en) * | 2014-12-26 | 2015-04-29 | 中国农业科学院棉花研究所 | Molecular identification method for zero-type fruit branch genes of cotton |
| WO2017009847A1 (en) * | 2015-07-16 | 2017-01-19 | Kaiima Bio Agritech Ltd. | Determinate castor |
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| CN103183732A (en) * | 2013-04-18 | 2013-07-03 | 中国农业科学院棉花研究所 | Cotton Gh FPP1 protein as well as coding gene and application thereof |
| CN103224555A (en) * | 2013-05-27 | 2013-07-31 | 中国农业科学院棉花研究所 | Plant-development-related protein GhSOC1, and coding gene and application thereof |
| CN104561284A (en) * | 2014-12-26 | 2015-04-29 | 中国农业科学院棉花研究所 | Molecular identification method for zero-type fruit branch genes of cotton |
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| CN112063741A (en) * | 2020-09-10 | 2020-12-11 | 中国农业科学院棉花研究所 | Cotton fruit node long and short molecular marker and application thereof |
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