CN107904245B - Application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice - Google Patents
Application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice Download PDFInfo
- Publication number
- CN107904245B CN107904245B CN201711252460.3A CN201711252460A CN107904245B CN 107904245 B CN107904245 B CN 107904245B CN 201711252460 A CN201711252460 A CN 201711252460A CN 107904245 B CN107904245 B CN 107904245B
- Authority
- CN
- China
- Prior art keywords
- zmnf
- rice
- seq
- transcription factor
- heading
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
-
- 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]
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Physiology (AREA)
- Plant Pathology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The application and its method that the present invention relates to Maize Transcription Factor ZmNF-YB2 in improvement rice are specially promoting application of the Rice Heading in advance or in reduction in the life period, and wherein the nucleotide sequence of ZmNF-YB2 gene is as shown in SEQ ID NO.4.By overexpression ZmNF-YB2 gene in rice, so that Rice Heading is in advance or reduction in the life period, this be that cultivation breeding time short rice varieties provide a method.
Description
Technical field
The invention belongs to field of biotechnology, are related to application of the Maize Transcription Factor ZmNF-YB2 in improvement rice, also
Be related to preparation heading in advance or reduction in the life period rice varieties method
Background technique
Field of plant genetic project technology achieves rapid progress in recent years.Firstly, turning BT gene pest-resistant cotton complete
The world is widely used, and the cost of applying pesticides is not only greatly saved, but also reduce the use of insecticide, to environment
Improvement play remarkable result, the kind of pest-resistant and anti-herbicide gene, also exists on the crops such as corn and soybean
It is used widely in production.The development of technique for gene engineering is also being constantly progressive the improvement of plant, such as phytase corn,
The phytase gene in microorganism is exactly transferred to corn, is being in this way original with corn so that corn seed be allowed to generate phytase
In the process of material production feed, the phosphorus in corn kernel is allowed to be released, without adding phosphorus nutrition element again, so not
Production cost is only reduced, the pollution waste of the exploitation of phosphorus ore is reduced, also reduces pollution of the animal excretion to environment.For another example, in water
In rice, by being transferred to the crtl gene from bacterium and the psy gene in corn, so that rice is allowed to contain carrotene,
Edible rice is solved to be deficient in vitamin the predicament of A.Simultaneously by transgenic approach, in crops such as corn, rice and wheats
The technology that disease-resistant performance, drought resistance and nutrition kind etc. improve is continuing to bring out.
Flowering In Higher Plant is solid be the growth and development period important link and reform of nature existence it is vital
Biological characteristics.Flowering of plant is not only controlled by heredity itself, also by the adjusting of the environmental factors such as ambient light, temperature.For rice
For equal crops, heading flowering time is directly related to the length of breeding time, is important economical character, determines its adaptation
Ecological range, and have great influence to final yield.
Summary of the invention
In view of this, one of the objects of the present invention is to provide a kind of Maize Transcription Factor ZmNF-YB2, and rice to be promoted to take out
Fringe shift to an earlier date or reduction in the life period in application；The second object of the present invention is to provide containing Maize Transcription Factor ZmNF-YB2 base
The recombinant vector or transformant of cause are promoting application of the Rice Heading in advance or in reduction in the life period；The third object of the present invention exists
In provide it is a kind of prepare heading in advance or reduction in the life period rice varieties method.
In order to achieve the above objectives, the invention provides the following technical scheme:
1, Maize Transcription Factor ZmNF-YB2 is promoting application of the Rice Heading in advance or in reduction in the life period, the corn
The gene nucleotide series of transcription factor ZmNF-YB2 are as shown in SEQ ID NO.4.
Preferably, the amino acid sequence of the Maize Transcription Factor ZmNF-YB2 gene coding is as shown in SEQ ID NO.5.
2, recombinant vector or transformant containing Maize Transcription Factor ZmNF-YB2 gene promoting Rice Heading in advance or
Application in reduction in the life period, the nucleotide sequence of the Maize Transcription Factor ZmNF-YB2 gene is as shown in SEQ ID NO.4.
3, it is a kind of prepare heading in advance or reduction in the life period rice varieties method, include the following steps:
1) using corn cDNA as template, sequence shown in SEQ ID NO.1 and SEQ ID NO.2 is that primer carries out PCR amplification,
Obtain nucleotide sequence Maize Transcription Factor ZmNF-YB2 gene as shown in SEQ ID NO.4；
2) the Maize Transcription Factor ZmNF-YB2 genetic fragment that amplification obtains is connected into through restriction enzyme Xcm I digestion
Plasmid pCXUN in, obtain the recombinant vector containing ZmNF-YB2 gene, then the recombinant vector containing ZmNF-YB2 is transferred to
Agrobacterium, screening positive clone obtain engineering bacteria；
3) by the engineering bacteria rice transformation of acquisition, screening positive plant, i.e. acquisition heading shift to an earlier date or the water of reduction in the life period
Rice.
Preferably, using corn RNA as template, cDNA is obtained through reverse transcription, then using cDNA as template, SEQ ID
Sequence shown in NO.1 and SEQ ID NO.2 is that primer carries out PCR amplification, obtains the corn of nucleotide sequence such as SEQ ID NO.4
Transcription factor ZmNF-YB2 gene.
It is furthermore preferred that the Agrobacterium is LBA4404.
Preferably, in step 3), the engineering bacteria rice transformation method is as follows: using agrobacterium-mediated transformation to pass through recombination
ZmNF-YB2 gene is transferred in the callus of Mature Embryos of Rice by carrier, callus after conversion by co-culturing, screening,
Differentiation, strong sprout and transplanting, screening transgenic plant, i.e. acquisition heading shift to an earlier date or the rice of reduction in the life period.
The beneficial effects of the present invention are: the invention discloses Maize Transcription Factor ZmNF-YB2 answering in rice breeding
With allowing Rice Heading to shift to an earlier date to realize, and shorten the breeding time of rice by overexpression ZmNF-YB2 gene in rice
7-10 days, there is important application value in the new rice variety for cultivating reduction in the life period.
Detailed description of the invention
In order to keep the purpose of the present invention, technical scheme and beneficial effects clearer, the present invention provides following attached drawing and carries out
Illustrate:
Fig. 1 is the corn ZmNF-YB2 genophore schematic diagram expressed in rice；
Fig. 2 is the PCR detection that ZmNF-YB2 gene is transferred to rice plant；
Fig. 3 is the PCR detection that ZmNF-YB2 gene is expressed in rice plant；
Fig. 4 is to turn ZmNF-YB2 trans-genetic hybrid rice and wild type control potting heading the case where blooming (004 is turns ZmNF-YB2
Rice, WT are Wild-type non-transgenic rice)；
Fig. 5 be turn ZmNF-YB2 trans-genetic hybrid rice and wild type control field ear situation (004 is turns ZmNF-YB2 rice,
WT is Wild-type non-transgenic rice).
Specific embodiment
The invention will now be further described with reference to specific embodiments, the features of the present invention will be with description and it is more clear
Chu.But the embodiment of description does not constitute any restrictions to protection scope of the present invention, and be only to be illustrated to technology
One example.It will be understood by those skilled in the art that without departing from the spirit of the invention based on this example, it can be to skill
The details and form of art scheme are modified or are replaced, but these modifications or substitutions all belong to the scope of protection of the present invention.
Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art,
Raw materials used is the commodity of market public offering.
Embodiment 1, Maize Transcription Factor ZmNF-YB2 Gene clone and expression vector construction
According to the result of study that Maize genome and transcript profile are sequenced, ZmNF-YB2 is selected in maize transcription group database
Gene designs upstream primer YB2-F:5 '-tgcaggaggagctcatggc-3 ' (SEQ ID NO.1), downstream primer YB2-R:
5'-caggtaagagcagctgtcgca-3'(SEQ ID NO.2).Corn " Hi II " blade total serum IgE of growth 20 days is extracted,
And as template, with Oligo (dT)18Reverse transcription is carried out for primer, obtains reverse transcription product cDNA.
The cDNA obtained using reverse transcription is template, with above-mentioned special primer SEQ ID NO.1 and SEQ ID NO.2 progress
PCR amplification obtains the DNA fragmentation of about 600bp, which includes ZmNF-YB2 gene, recycles target fragment, spare.
The method cloned using TA, the DNA fragmentation of the recycling is directly connected to and is obtained through restriction enzyme XcmI digestion
The carrier pCXUN segment arrived connects mixture Transformed E .coli DH5 α competent cell, selects the bacterium colony culture of normal growth,
The Plasmid DNA for extracting bacterium colony, carries out PCR with primer YB2-F and YB2-R, obtains the genetic fragment of equally about 600bp.Positive gram
It is grand further with sequencing primer: NosR-seq:5'-agaccggcaacaggattcaatc-3'(SEQ ID NO.3) carry out DNA
Sequencing is compared analysis and obtains the clone for containing correct ZmNF-YB2 gene, as constructed successful expression vector, order
Entitled pCXUN-ZmNF-YB2, structure are as shown in Figure 1.The nucleotide sequence of ZmNF-YB2 gene such as SEQ ID in the expression vector
Shown in NO.4, the amino acid sequence of translated protein is as shown in SEQ ID NO.5.
Embodiment 2, the preparation for turning ZmNF-YB2 trans-genetic hybrid rice
(1) preparation of the engineering bacteria of the expression vector containing pCXUN-ZmNF-YB2
The Agrobacterium LBA4404 of -80 DEG C of preservations is taken to line LB (containing 50mg/L rifampin) plate, 28 DEG C are cultivated 2 days.
Single colonie is chosen on the plate turned out, is inoculated in 50mL liquid LB (rifampin containing 50mg/L) culture, the 150rpm at 28 DEG C
Shaking table vibration is cultivated to OD600=0.5；Then Agrobacterium bacterium solution is transferred to sterile 50ml centrifuge tube, 4 DEG C, 4000rpm is centrifuged
10min abandons supernatant, 5mL is added, 0.15M NaCl suspended bacterial cell is pre-chilled on ice water, 4 DEG C, 4000rpm is centrifuged
10min abandons supernatant；0.15M NaCl suspension cell is used again, and 4 DEG C, 4000rpm is centrifuged 10min, abandons supernatant；It is pre- that 2mL is added
Cold 20mmol/L CaCl2Solution, gently suspension cell, is placed on ice, final concentration of 7% (V/V) DMSO is added, after mixing
Packing, every 100 μ L of pipe；It is frozen immediately after liquid nitrogen frozen is added in -80 DEG C of low temperature refrigerators.
The pCXUN-ZmNF-YB2 expression vector of 1 μ g or so is taken to be added in 100 μ L Agrobacterium competent cells, after mixing
It is placed in 30min on ice；Liquid nitrogen frozen 1min, in 37 DEG C of water-baths to thawing after taking-up, liquid nitrogen frozen 1min again immediately after defrosting,
37 DEG C of water-baths are placed in ice water 2min to thawing after taking-up；1mL LB culture medium is added, the 140rpm shaking table vibration culture at 28 DEG C
2-3h；It is centrifugated thallus, 100 μ l LB liquid mediums are added and are resuspended, are coated in+50mg/L the strepto- of kanamycins containing 100mg/L
On the LB plate of element, 28 DEG C of culture to single colonies are formed；Single bacterium colony is inoculated in the liquid LB containing 100mg/L kanamycins,
140rpm shaking table vibration culture 16h, takes 2 μ L bacterium solutions to carry out PCR detection at 28 DEG C, and positive colony is saved backup at -80 DEG C.
(2) Agrobacterium-Mediated Transformation in Oryza sativa
The Agrobacterium LBA4404 positive bacterium colony with ZmNF-YB2 target gene plasmid is picked from the plate, is inoculated in
The flat lining out of LB+50mg/L streptomysin+100mg/L kanamycins, 19 DEG C are cultivated three days.It is suspended with a small amount of bacterium of oese picking
The dip dyeing culture medium of 100 μm of ol/L acetosyringones is added in 15mL, bacterial concentration is controlled in OD550=0.06-0.08.This bacterium
Liquid is the agrobacterium suspension of rice transformation.
Water intaking rice " Kasalath " mature seed decladding, the seed for selecting full bright and clean no bacterial plaque is sterilized, is then inoculated with
Evoked callus is carried out on to induced medium；Select appearance good, eugonic callus is transformation receptor material,
Callus 2min is impregnated with above-mentioned agrobacterium liquid, then this callus impregnated is placed on co-cultivation base and is total to
It cultivates, is transferred on screening and culturing medium and is cultivated twice after 19 DEG C of dark culture 3d, each 14d subculture；The kanamycin-resistant callus tissue sifted out is shifted
To differential medium, differentiation culture differentiated seedling after about 14 days one by one；The green seedling differentiated is transferred to life
It takes root on root culture medium, the hardening after about 20d grows flourishing root system；Grown in field cultivation is transferred to after hardening 3-5d.
Culture medium prescription: N6+0.7g/L L-PROLINE+68.4g/L sucrose+36g/L glucose+1.5mg/L 2 is disseminated,
4-D, pH5.2.
Fiber differentiation based formulas: N6+300mg/L caseinhydrolysate+2.8g/L L-PROLINE+30g/L sucrose+2mg/L2,
4-D+4g/L plant gel, pH5.8.
Co-culture based formulas are as follows: the Portugal N6+300mg/L caseinhydrolysate+2.8g/L L-PROLINE+30g/L sucrose+10g/L
Grape sugar+2mg/L 2,4-D+100 μm of ol/L acetosyringone+4g/L plant gel, pH5.8.
Screening and culturing based formulas are as follows: N6+300mg/L caseinhydrolysate+2.8g/L L-PROLINE+30g/L sucrose+2mg/
L2,4-D+4g/L plant gel+50mg/L hygromycin+250mg/L carbenicillin, pH5.8.
Differential medium formula are as follows: MS+2g/L caseinhydrolysate+30g/L sucrose+30g/L sorbierite+2.0mg/L excitement
Element+0.02mg/L niacin+4g/L plant gel+50mg/L hygromycin+100mg/L cephalosporin+10mg/L vancomycin,
pH5.8。
Prescription of rooting medium are as follows: 1/2MS+20g/L sucrose+0.5mg/L niacin+4g/L plant gel, pH5.8.
The PCR detection of embodiment 3, transgenosis
ZmNF-YB2 gene whether is transferred in embodiment 2 in rice conversion plant in order to detect, using on Vector promoter
A primer Ubi-F:5 '-ttttagccctgccttcatacgc-3 ' (SEQ ID NO.6) and transgene ZmNF-YB2
Downstream primer SEQ ID NO.2 carries out PCR detection, is expanded using the DNA of rice transformation plant as template, and transgenic positive is planted
Strain can expand to obtain the DNA band that clip size is more than 650 bp or so, rather than transgenic paddy rice can not expand to obtain this
Band.PCR reacts 50 μ L of total volume, illustrates to prepare by manufacturer.PCR amplification program is as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation
30sec, 58 DEG C of annealing 30sec, 72 DEG C of extension 45sec, 34 recycle；72 DEG C of extension 6min.PCR product is solidifying through 1% agarose
Gel electrophoresis observation, as a result as shown in Figure 2.
Embodiment 4 turns the PCR detection that ZmNF-YB2 gene is expressed in rice plant
In order to detect ZmNF-YB2 gene in T2 for the expression in transgenic paddy rice, using PCR method to its
It is identified on transcriptional level, as a result as shown in Figure 3.The result shows that transgenic plant can detect the expression of the gene, and it is wild
Raw type plant can't detect.
The process of PCR experiment detection is as follows: taking appropriate transgenic line and wild rice blade, is mentioned using TRIzol method
Total serum IgE is taken, then reverse transcription is cDNA, carries out PCR reaction, PCR the primer sequence are as follows: 5 '-as template
Tgcaggaggagctcatggc-3 ' (SEQ ID NO.1), downstream primer NosR-seq:5'-
Agaccggcaacaggattcaatc-3'(SEQ ID NO.3), transgenic positive plant can expand to obtain clip size be
The DNA band of more than 650 bp or so, rather than transgenic paddy rice can not expand to obtain this band.PCR reacts 50 μ L of total volume, by factory
Quotient illustrates to prepare.PCR amplification program is as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30sec, 58 DEG C of annealing 30sec, 72 DEG C are prolonged
Stretch 45sec, 34 circulations；72 DEG C of extension 6min；PCR product is observed through 1% agarose gel electrophoresis.
Embodiment 5 turns ZmNF-YB2 trans-genetic hybrid rice phenotypic analysis
ZmNF-YB2 transgenic paddy rice and wild rice ' Kasalath ' are observed, as a result as shown in Figure 4 and Figure 5.With open country
Raw type rice ' Kasalath ' is compared, and the heading stage of ZmNF-YB2 transgenic paddy rice obviously shifts to an earlier date, and about in advance 7-10 days or so.Such as
Shown in Fig. 5, wild type just starts to ear, but transgenic line is basically completed heading.Economical character investigates result such as 1 institute of table
Show, investigates result and tentatively show to turn ZmNF-YB2 trans-genetic hybrid rice partially short more than 10 cm generally than non-transgenic control Plant Height of Rice, often
Fringe bear fruit grains relatively compare it is on the low side, but number of productive ear relatively control generally increased, spike length and mass of 1000 kernel variation it is unobvious, and survey produce
Turn ZmNF-YB2 trans-genetic hybrid rice and control when amount without significant difference.
1. turns of ZmNF-YB2 plant of table and nontransgenic plants economical character application form
004A, 004B, 004C: for the different transformation plant of ZmNF-YB2；WT: for nontransgenic plants；*Indicate transgenosis
Level of signifiance P < 0.05 compared between strain and non-transgenic strain WT.
To sum up the result shows that being overexpressed ZmNF-YB2 gene in rice, remarkably promotes rice ear sprouting period and shift to an earlier date, about shift to an earlier date
7-10 days, although plant height and fringe bear fruit grains decrease, number of productive ear was increased, and it is unaffected to eventually lead to yield.This
Cultivating has important application value in the new rice variety of reduction in the life period.
Sequence table
<110>Institute of Nuclear and Biotechnology, Sichuan Academy of Agriculture Science
<120>application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
tgcaggagga gctcatggc 19
<210> 2
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
caggtaagag cagctgtcgc a 21
<210> 3
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
agaccggcaa caggattcaa tc 22
<210> 4
<211> 591
<212> DNA
<213>corn (Zea mays)
<400> 4
tgcaggagga gctcatggcg gaagctccgg cgagccctgg cggcggcggc gggagccacg 60
agagcgggag ccccagggga ggcggaggcg gtggcagcgt cagggagcag gacaggttcc 120
tgcccatcgc caacatcagt cgcatcatga agaaggccat cccggctaac gggaagatcg 180
ccaaggacgc taaggagacc gtgcaggagt gcgtctccga gttcatctcc ttcatcacta 240
gcgaagcgag tgacaagtgc cagagggaga agcggaagac catcaatggc gacgatctgc 300
tgtgggccat ggccacgctg gggtttgaag actacattga acccctcaag gtgtacctgc 360
agaagtacag agagatggag ggtgatagca agttaactgc aaaatctagc gatggctcaa 420
ttaaaaagga tgcccttggt catgtgggag caagtagctc agctgcacaa gggatgggcc 480
aacagggagc atacaaccaa ggaatgggtt atatgcaacc ccagtaccat aacggggata 540
tctcaaacta atggaggtat ggaccttttc tgcgacagct gctcttacct g 591
<210> 5
<211> 178
<212> PRT
<213>corn (Zea mays)
<400> 5
Met Ala Glu Ala Pro Ala Ser Pro Gly Gly Gly Gly Gly Ser His Glu
1 5 10 15
Ser Gly Ser Pro Arg Gly Gly Gly Gly Gly Gly Ser Val Arg Glu Gln
20 25 30
Asp Arg Phe Leu Pro Ile Ala Asn Ile Ser Arg Ile Met Lys Lys Ala
35 40 45
Ile Pro Ala Asn Gly Lys Ile Ala Lys Asp Ala Lys Glu Thr Val Gln
50 55 60
Glu Cys Val Ser Glu Phe Ile Ser Phe Ile Thr Ser Glu Ala Ser Asp
65 70 75 80
Lys Cys Gln Arg Glu Lys Arg Lys Thr Ile Asn Gly Asp Asp Leu Leu
85 90 95
Trp Ala Met Ala Thr Leu Gly Phe Glu Asp Tyr Ile Glu Pro Leu Lys
100 105 110
Val Tyr Leu Gln Lys Tyr Arg Glu Met Glu Gly Asp Ser Lys Leu Thr
115 120 125
Ala Lys Ser Ser Asp Gly Ser Ile Lys Lys Asp Ala Leu Gly His Val
130 135 140
Gly Ala Ser Ser Ser Ala Ala Gln Gly Met Gly Gln Gln Gly Ala Tyr
145 150 155 160
Asn Gln Gly Met Gly Tyr Met Gln Pro Gln Tyr His Asn Gly Asp Ile
165 170 175
Ser Asn
<210> 6
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 6
ttttagccct gccttcatac gc 22
Claims (7)
1. Maize Transcription Factor ZmNF-YB2 is promoting application of the Rice Heading in advance or in reduction in the life period, it is characterised in that:
The gene nucleotide series of the Maize Transcription Factor ZmNF-YB2 are as shown in SEQ ID NO.4.
2. Maize Transcription Factor ZmNF-YB2 is promoting application of the Rice Heading in advance or in reduction in the life period, it is characterised in that:
The amino acid sequence of the gene coding of the Maize Transcription Factor ZmNF-YB2 is as shown in SEQ ID NO.5.
3. recombinant vector or transformant containing Maize Transcription Factor ZmNF-YB2 gene are promoting Rice Heading in advance or fertility
Application in phase shortening, it is characterised in that: the nucleotide sequence of the Maize Transcription Factor ZmNF-YB2 gene such as SEQ ID
Shown in NO.4.
4. it is a kind of prepare heading in advance or reduction in the life period rice varieties method, which comprises the steps of:
1) using corn cDNA as template, sequence shown in SEQ ID NO.1 and SEQ ID NO.2 is that primer carries out PCR amplification, is obtained
Nucleotide sequence Maize Transcription Factor ZmNF-YB2 gene as shown in SEQ ID NO.4；
2) the Maize Transcription Factor ZmNF-YB2 genetic fragment that amplification obtains is connected into the matter through restriction enzyme Xcm I digestion
In grain pCXUN, the recombinant vector containing ZmNF-YB2 gene is obtained, the recombinant vector containing ZmNF-YB2 is then transferred to agriculture bar
Bacterium, screening positive clone obtain engineering bacteria；
3) by the engineering bacteria rice transformation of acquisition, screening positive plant, i.e. acquisition heading shift to an earlier date or the rice of reduction in the life period.
5. prepare according to claim 4 heading in advance or reduction in the life period rice varieties method, it is characterised in that: step
It is rapid 1) are as follows: using corn RNA as template, obtain cDNA through reverse transcription, then using cDNA as template, SEQ ID NO.1 and SEQ
Sequence shown in ID NO.2 is that primer carries out PCR amplification, obtains the Maize Transcription Factor of nucleotide sequence such as SEQ ID NO.4
ZmNF-YB2 gene.
6. prepare according to claim 4 heading in advance or reduction in the life period rice varieties method, it is characterised in that: step
It is rapid 2) in, the Agrobacterium be LBA4404.
7. prepare according to claim 4 heading in advance or reduction in the life period rice varieties method, it is characterised in that: step
It is rapid 3) in, the engineering bacteria rice transformation method is as follows: use agrobacterium-mediated transformation for by recombinant vector by ZmNF-YB2 base
Because being transferred in the callus of Mature Embryos of Rice induction, callus after conversion by co-culturing, screening, differentiation, strong sprout and
Transplanting, screening transgenic plant, i.e. acquisition heading shift to an earlier date or the rice of reduction in the life period.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711252460.3A CN107904245B (en) | 2017-12-01 | 2017-12-01 | Application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711252460.3A CN107904245B (en) | 2017-12-01 | 2017-12-01 | Application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107904245A CN107904245A (en) | 2018-04-13 |
| CN107904245B true CN107904245B (en) | 2019-09-24 |
Family
ID=61849673
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711252460.3A Active CN107904245B (en) | 2017-12-01 | 2017-12-01 | Application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107904245B (en) |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103265634B (en) * | 2013-04-28 | 2014-11-26 | 中国农业科学院作物科学研究所 | Application of rice transcription factor Os01g64730 gene |
| CN103305530A (en) * | 2013-05-29 | 2013-09-18 | 四川农业大学 | Corn transcription factor zmwrky44 and application thereof |
| CN104498505A (en) * | 2014-12-09 | 2015-04-08 | 四川省农业科学院生物技术核技术研究所 | Application of gene AtNF-YB-1 in promotion of early head sprouting and short growth period of plant |
| CN104593411B (en) * | 2015-02-15 | 2017-08-29 | 福建省农业科学院生物技术研究所 | Application of the barley Hvsusiba2 genes in reduction rice terrace methane release |
| CN107298701B (en) * | 2017-04-18 | 2020-10-30 | 上海大学 | Corn transcription factor ZmbZIP22 and application thereof |
| CN107058343B (en) * | 2017-06-21 | 2020-12-29 | 四川农业大学 | Application of transcription factor VP1 in regulation of plant height and grain starch content of crops |
-
2017
- 2017-12-01 CN CN201711252460.3A patent/CN107904245B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN107904245A (en) | 2018-04-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110669785B (en) | Application of tomato SlLOB40 protein and coding gene thereof in regulation and control of plant drought resistance | |
| CN101743314A (en) | Transgenic plants with increased stress tolerance and yield | |
| CN104611359B (en) | The application of ZmSPL1 albumen and its encoding gene in regulation and control Maize Kernel Development | |
| CN107058343B (en) | Application of transcription factor VP1 in regulation of plant height and grain starch content of crops | |
| CN107164391A (en) | A kind of strawberry floral genes FvbHLH78 and its application | |
| CN112779234B (en) | Phyllostachys pubescens PeAPX5 gene and application thereof | |
| CN112725349B (en) | Dactylis glomerata zinc finger protein gene DgMYM1, and expression vector and application thereof | |
| CN110066774A (en) | Corn receptoroid kinase gene ZmRLK7 and its application | |
| CN108570099A (en) | The application of OsGLP2-1 albumen and its encoding gene in regulating and controlling seed dormancy | |
| CN114214358B (en) | Inducible expression vector and application thereof in culturing sentry crops | |
| CN107815452A (en) | A kind of specific expressed promoter of plant leaf blade and its application | |
| CN110904106B (en) | Application of cymbidium goeringii miR159b in enhancing plant cold sensitivity | |
| CN109096380B (en) | Application of OsBICs gene in regulation and control of plant height and flowering time | |
| CN109485710B (en) | Rice seed dormancy regulatory gene OsAnn3, protein coded by same and application of gene OsAnn3 | |
| CN107904245B (en) | Application and its method of the Maize Transcription Factor ZmNF-YB2 in improvement rice | |
| CN112795580B (en) | Pitaya gene HuAAE3 and application thereof in regulation and control of high temperature stress resistance of plants | |
| CN112458101B (en) | Duck grass zinc finger protein gene DgZFP125 as well as expression vector and application thereof | |
| CN107973844A (en) | Wheat heading stage GAP-associated protein GAP Ta-Hd4A and its application | |
| CN110343159B (en) | Application of expression vector of mung bean flowering gene VrELF3 | |
| CN111304198B (en) | Application of cymbidium goeringii miR390b in controlling plant vegetative organ development | |
| CN110387376B (en) | Application of expression vector containing mung bean flowering gene VrFT5a | |
| CN104561040B (en) | Genes For Plant Tolerance hot radical is because of HTT3 and its application | |
| KR101855137B1 (en) | ATPG8 Protein Providing Yield Increase and Delaying Senescence and Stress Tolerance of Plants, the Gene Encoding the Protein and Those Use | |
| CN103665129A (en) | Plant heading period associated protein TaMYB72 and application thereof | |
| CN114015666B (en) | Application of OsPARP3 gene in regulation and control of plant drought tolerance |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |