CN114672511A - Application of corn ZmBES1/BZR1-3 gene in increasing plant seed yield - Google Patents
Application of corn ZmBES1/BZR1-3 gene in increasing plant seed yield Download PDFInfo
- Publication number
- CN114672511A CN114672511A CN202210270506.9A CN202210270506A CN114672511A CN 114672511 A CN114672511 A CN 114672511A CN 202210270506 A CN202210270506 A CN 202210270506A CN 114672511 A CN114672511 A CN 114672511A
- Authority
- CN
- China
- Prior art keywords
- bzr1
- zmbes1
- corn
- gene
- genes
- 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.)
- Granted
Links
Images
Classifications
-
- 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
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
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)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses application of a corn ZmBES1/BZR1-3 gene in increasing plant seed yield, wherein the corn ZmBES1/BZR1-3 gene is cloned by using corn seedling leaves, and the function of the gene is researched from the phenotypic characteristics of transgenic arabidopsis thaliana and rice, so that a new choice is provided for high-yield breeding of rice, improvement of economic benefit of rice and the like.
Description
Technical Field
The invention relates to application of a corn ZmBES1/BZR1-3 gene in increasing the yield of plant seeds, belonging to the field of genetic engineering.
Background
Grain yield is always important content concerned by people, and the improvement of grain yield has great significance to economic development and grain safety in China. The yield is a complex quantitative character and is influenced by various factors such as plant type, panicle grain, growth period and the like. The rice is a main grain crop, the yield of a single rice plant is mainly determined by the characters such as the effective ear number, the grain number per ear, the thousand seed weight and the like, the thousand seed weight is the character with the highest heritability in yield composition factors and is determined by the grain shape and the filling degree, and the grain shape is determined by the grain length, the grain width and the grain thickness. In the prior art, mainly some rice self-existing genes are applied to the rice yield, GS3 is the first main effect QTL of grain length and grain weight obtained by a map-based cloning method, has weak effects on grain width and grain thickness, and simultaneously, GS3 protein is found to contain a plant-specific organ size control structural domain (OSR), and large grains are formed after the structural domain is deleted or mutated, so that the size of the OSR negative control grain shape is deduced. DEP1 is also a subunit of the G protein and affects not only spike shape but also granule shape. QTL GL7/GW7 encodes a LONGIFOLIA protein, and up-regulation of expression of GW7 can slow down transverse cell division and promote the formation of long and thin grains. qSW5/GW5 is a gene controlling rice grain width and grain weight on rice chromosome 5, and GW5 can interact with polyubiquitin to regulate grain width and grain weight through ubiquitin proteasome pathway. GW2 negatively regulates rice grain width, and increasing the expression of GW2 can result in narrowing seed grain width.
In recent years, the rice yield is gradually improved by introducing a single corn gene into a rice plant, the rice yield is generally improved by improving the photosynthetic rate of rice, C4 photosynthetic related enzyme is introduced into C3 crops such as rice, for example, PEPCase is a key enzyme of a C4 pathway, the corn PEPC gene is successfully transferred into the rice, the CO2 compensation point and the light respiration rate of the transgenic rice are obviously reduced, the apparent net photosynthetic rate is improved, and the yield is also improved to different degrees. And relatively little research has been conducted on genes other than photosynthesis.
Disclosure of Invention
The invention overcomes the defects of the prior art and provides the application of the corn ZmBES1/BZR1-3 gene in increasing the yield of plant seeds, the corn ZmBES1/BZR1-3 gene is cloned by utilizing the leaves of corn seedlings, the function of the gene is researched in the aspect of the phenotypic characteristics of transgenic arabidopsis thaliana and rice, and a new choice is provided for high-yield breeding of rice, improvement of the economic benefit of the rice and the like.
The application of the corn ZmBES1/BZR1-3 gene in increasing the seed yield of plants, wherein the nucleotide sequence of the corn ZmBES1/BZR1-3 gene is shown in SED ID NO. 1.
The application of the corn ZmBES1/BZR1-3 gene in improving the phenotype of plant seeds, wherein the nucleotide sequence of the corn ZmBES1/BZR1-3 gene is shown in SED ID NO. 1.
Further, the above plant seed phenotype refers to seed length, seed width and thousand seed weight.
Further, the above plant refers to Arabidopsis thaliana or rice.
A method for cultivating high-yield plants comprises the following steps: the expression of corn ZmBES1/BZR1-3 genes in the plant is up-regulated to obtain a plant with over-expressed ZmBES1/BZR1-3 genes, thereby obtaining a high-yield plant.
Further, the method comprises the following specific steps: separating corn ZmBES1/BZR1-3 genes from corn seedling leaves, designing amplification primers of the corn ZmBES1/BZR1-3 genes, amplifying cDNA sequences of the corn ZmBES1/BZR1-3 genes by a PCR method, constructing a bacterium-mediated 35S-ZBES 1/BZR1-3-eGFP vector by using the cDNA sequences of the ZmBES1/BZR1-3, transforming the vector into a target plant by using an agricultural rod method, improving the expression of the ZmBES1/BZR1-3 genes, and obtaining a plant with over-expressed ZmBES1/BZR1-3 genes, thereby obtaining a high-yield plant.
Furthermore, the nucleotide sequence of the maize ZmBES1/BZR1-3 gene is shown as SED ID NO. 1.
Furthermore, the nucleotide sequences of the amplification primers of the maize ZmBES1/BZR1-3 gene are shown as SED ID NO.2 and SED ID NO. 3.
Further, the nucleotide sequence of the above 35S-ZmBES1/BZR1-3-eGFP vector is shown as SED ID NO. 10.
Further, the above plant refers to Arabidopsis thaliana or rice.
Has the beneficial effects that:
the invention provides a method for improving the yield of arabidopsis thaliana and rice seeds by using corn ZmBES1/BZR1-3 genes for the first time, the method improves the yield of arabidopsis thaliana or rice by improving the expression of the corn ZmBES1/BZR1-3 genes, provides a new molecular breeding approach for culturing high-yield rice fine varieties, and has wide application prospect. The invention can obviously increase the grain width, the grain length and the thousand seed weight of arabidopsis or rice plants.
Drawings
FIG. 1 homozygous transgenic Arabidopsis seed phenotype.
FIG. 2 transgenic Arabidopsis seeds are long and wide.
FIG. 3 transgenic rice seed phenotype.
FIG. 4 shows the length and width of the transgenic rice seed.
FIG. 5 shows thousand kernel weight of homozygous transgenic rice.
Detailed Description
In order to make the technical solutions in the present application better understood, the present invention is further described below with reference to examples, which are only a part of examples of the present application, but not all examples, and the present invention is not limited by the following examples.
Examples
1. Gene cloning and vector construction
Taking five-leaf stage corn B73 seedling leaves, carrying out quick-freezing grinding by liquid nitrogen, and extracting total RNA by using a total RNA extraction kit Trizol (TaKaRa) according to the instruction. Using PrimeScript TMRT regant Kit (Takara) was used to synthesize cDNA using total RNA as a template. Clone primers were designed using Primer 5.0, using B73 cDNA as template, and specific PCR primers are shown in Table 1. Amplification was performed using high fidelity enzyme PrimerStar (Takara Inc.), PCR amplification reaction system as shown in Table 2, temperature cycling program: 94 ℃ for 3 min; 98 ℃, 10s and the optimum annealing temperature of 10 s; 72 ℃ for 20 s; 30 cycles; 5min at 72 ℃; keeping at 4 ℃.
TABLE 1 ZmBES1/BZR1-3 cloning primer
TABLE 2 PCR amplification reaction System
1.1 transformation of Arabidopsis thaliana vector 35S-ZmBES1/BZR1-3-eGFP construction
According to the dicotyledonous plant expression vector pCAMBIA2300-35S-eGFP multiple cloning site, an upstream primer thereof enters a BamH I enzyme cutting site, and a downstream primer thereof is introduced into a Sal I site, and the specific sequence is as follows:
table 3 construction of pCAMBIA2300-35S-eGFP vector primers
Samples were added to the reaction system shown in Table 1 and amplification was carried out. The temperature cycling program was: 3min at 94 ℃; 10s at 98 ℃, 10s at the optimum annealing temperature, 20s at 72 ℃ and 30 cycles; 5min at 72 ℃; keeping at 4 ℃.
And recovering the PCR product. And carrying out double enzyme digestion on the recovered product, namely pCAMBIA2300-35S-eGFP plasmid, and loading the sample according to a surface reaction system. Uniformly mixing the enzyme digestion system, placing the mixture in a water bath kettle at 30 ℃, carrying out enzyme digestion for 6-8 h, adding the enzyme digestion product into a loading buffer solution, carrying out direct electrophoresis, recovering the enzyme digestion product, using a Cloneexpress II One Step Cloning Kit (nunoprazan), wherein the reaction system is shown in Table 5, and the reaction conditions are as follows: 30mins at 37 ℃.
TABLE 4 double digestion reaction System
TABLE 5 connection System
The ligation product was transformed into E.coli and tested by colony PCR. After the extracted recombinant plasmid is subjected to restriction enzyme digestion identification, the plasmid-composed bacterial liquid is sent to a sequencing company for DNA sequencing identification, and the plasmid with correct sequencing is converted into arabidopsis thaliana in the next step.
1.2 construction of transformed Rice vector
According to the monocotyledon expression vector pCAMBIA1300-35S-eGFP multiple cloning site, a Hind III enzyme cutting site is introduced into the upstream primer of the cloning site, and a BamH I site is introduced into the downstream primer, wherein the specific sequence is shown in a table 6. Amplification was performed using high fidelity enzyme PrimerStar (Takara Co.) and the PCR amplification reaction system is shown in Table 2. The PCR product is then recovered.
TABLE 6 construction of pCAMBIA1300-35S-ZmBES1/BZR1-3-eGFP vector primers
The expression vector pCAMBIA1300-35S-eGFP plasmid was digested simultaneously with the reaction system shown in Table 5 using Clonexpress II One Step Cloning Kit (Novozam), and the reaction conditions were as follows: 30mins at 37 ℃.
Table 44 the reaction system was loaded. Uniformly mixing the enzyme digestion system, placing the mixture in a water bath kettle at 30 ℃, carrying out enzyme digestion for 6-8 h, adding the enzyme digestion product into a loading buffer solution, carrying out direct electrophoresis, recovering the enzyme digestion product, using a Cloneexpress II One Step Cloning Kit (nunoprazan), wherein the reaction system is shown in Table 5, and the reaction conditions are as follows: 30mins at 37 ℃.
The ligation product was transformed into E.coli and tested by colony PCR. After the extracted recombinant plasmid is subjected to restriction enzyme digestion identification, the plasmid-composed bacterial liquid is sent to a sequencing company for DNA sequence sequencing identification, and the plasmid with correct sequencing is converted into rice in the next step.
2. Arabidopsis and rice transformation and positive identification
2.1 transformation of Arabidopsis thaliana by Flowery infection
Exploring the function of maize ZmBES1/BZR1-3, we transformed the Arabidopsis BES1 mutant (BES1-D) with the pCAMBIA2300-35S-ZmBES1/BZR1-3-eGFP vector using Agrobacterium-mediated catoptric floral dip.
2.1.1 inflorescence Dip-dyeing transformation
(1) Taking agrobacterium containing the recombinant plasmid, streaking on a YEP plate containing Kana and Rif, and culturing for 2-3 d at 28 ℃.
(2) A single colony of Agrobacterium was picked and inoculated in 3mL of YEP liquid medium containing Kana and Rif, cultured at 28 ℃ at 200r/min, and shaken overnight.
(3) Inoculating 1mL of overnight-cultured starting Agrobacterium strain in 100mL of liquid YEP medium (containing Kana and Rif), and shake-culturing at 28 deg.C to OD600The value is 1.2 to 1.5.
(4) Centrifuging at 4 deg.C for 10min at 5000r/min to collect cells, suspending thallus with 5% sucrose solution and adjusting OD600When the value is 0.8-1.0, the surfactant silwet L-77 is added according to the proportion of 1-2%.
(5) And (3) carrying out pot culture on the Arabidopsis BES1 mutant by using special nutrient soil for Arabidopsis, pruning formed pods and opened flowers after flowering, soaking inflorescences for 1.5-2.0 min by using the staining solution, and carrying out dark culture for 10 h.
(6) Culturing for 3-4 weeks under appropriate conditions, collecting seeds, and performing the next screening treatment.
2.1.2 transgenic line screening
(1) Harvested infected seeds were aliquoted into 1.5mL EP tubes and soaked in 500. mu.L of 70% ethanol for 30 s.
(2) After the alcohol is sucked off, soaking the seeds in 500 mu L of 10% sodium hypochlorite for 5-10 min, and continuously shaking the centrifugal tube during the soaking process until the seeds are fully contacted with the sodium hypochlorite.
(3) Washing the seeds with sterilized water for 4-6 times, and abandoning the upper ddH layer after the seeds are settled2O; 200 μ L of 0.1% agar water suspend seeds.
(4) The seeds were sown on 1/2MS medium containing 40mg/L kanamycin (Kana) and cultured.
(5) Repeating the above steps to harvest the transformed Arabidopsis thaliana T2After seed generation disinfection, the seeds are sown on 1/2MS solid culture medium containing kanamycin, after 7-8 days, all green robust plants are homozygous transformants, and non-homozygous transformants show 3:1 segregation (proportion of normal growth and etiolating dead plants). Laying downIs homozygous for T 3The seed generations were used for further analysis.
2.2. Transformation of rice
The constructed vector is sent to the Nanomi Bio Inc. (Nanjing) to transform the wild type Nippon (NIP) of the rice.
2.2.1 genomic PCR identification
Performing DNA extraction and PCR identification on the obtained positive strains
(1) Taking 100mg of fresh plant tissue, adding liquid nitrogen, and fully grinding. 400 μ L of buffer FP1 and 6 μ L of RNaseA (10mg/mL) were added, vortexed for 1min, and allowed to stand at room temperature for 10 min.
(2) Add 130. mu.L of buffer FP2, mix well, vortex for 1min, centrifuge at 12000r/min (13400 Xg) for 5min, transfer the supernatant to a new centrifuge tube.
(3) Optional steps are as follows: the supernatant was centrifuged again at 12000r/min (13400 Xg) for 5min and transferred to a new centrifuge tube.
(4) 0.7 times volume of isopropanol is added into the supernatant, and the mixture is fully mixed, so that flocculent genomic DNA can appear. The mixture was centrifuged at 12000r/min (13400 Xg) for 2min, the supernatant was discarded, and the precipitate was retained.
(5) Add 600. mu.L 70% ethanol, vortex for 5s, centrifuge at 12000r/min (13400 Xg) for 2min, and discard the supernatant.
(6) And (6) repeating the step.
(7) And (5) opening the cover and inverting the cover, and completely airing the residual ethanol at room temperature for 5-10 min.
(8) Adding a proper amount of elution buffer TE, dissolving DNA in water bath at 65 ℃ for 10-60 min, and reversely and uniformly mixing for several times to aid dissolution to finally obtain a DNA solution.
(9) PCR detection was performed using PCR primers as shown in Table 8, using the extracted genomic DNA as template, with the program 94 ℃ for 3 min; 94 ℃ for 30s, the optimum annealing temperature for 30s, 72 ℃ for 60s/kb, 35 cycles; 5min at 72 ℃; storing at 4 ℃.
TABLE 7 transgenic Rice detection primers
(10) Harvesting of transformed Rice seed T2Generation, seeds of all positive lines after sowing were examined for progeny for further analysis.
3. Analysis of transgenic line yield
3.1 Arabidopsis seed phenotypic characterization
Homozygous transgenic arabidopsis lines L3-1, L3-10 and mutant (bes1-D) were sterilized with 5% sodium hypochlorite solution, sown on medium containing 1/2MS, and 15D later transplanted to nutrient soil: in a 50mm x 50mm pot containing 3:1 soil of vermiculite, 4 plants were sown in each pot, and the pot was cultivated under the same conditions for direct harvest, and the seed length and width were measured.
3.2 phenotypic identification of Rice seeds
Homozygous transgenic rice lines L3-1 and L3-10 and wild type (NIP) were sown in paddy fields, straight-harvested seeds were cultured under the same conditions, and seed grain length, grain width and thousand kernel weight were measured.
4. Analysis of transgenic line yield
4.1 transgenic Arabidopsis yield analysis
Through resistance screening, the target gene ZmBES1/BZR1-3 is determined to be stably expressed in arabidopsis, and the seed grain length and the grain width of homozygous transgenic arabidopsis are analyzed (figure 1), and the result shows that the grain length of seeds of an over-expression strain L3-10 is remarkably increased, and the grain widths of seeds of L3-1 and L3-10 are remarkably increased (figure 2, wherein the x and the x respectively represent that p is less than 0.05 and p is less than 0.01).
4.2 transgenic Rice yield analysis
And detecting the target gene by PCR to determine a positive strain. Analysis of the positive lines for grain length, grain width and thousand grain weight (fig. 3) showed a very significant increase in grain length with no significant change in grain width for the over-expressed lines R3-3 and R3-4 (fig. 4, x and x indicate p <0.05, p <0.01, respectively). Thousand kernel weight was significantly increased compared to control NIP (figure 5, with indicates p <0.05, p <0.01, respectively).
SEQUENCE LISTING
<110> Sichuan university of agriculture
Application of <120> corn ZmBES1/BZR1-3 gene in increasing yield of plant seeds
<130> 2022
<160> 10
<170> PatentIn version 3.3
<210> 1
<211> 1311
<212> DNA
<213> sequence
<400> 1
atggagggag gggtaggagc gggaggaagt cggggtgatg agggttgggc tagggcagga 60
gacggggacg ggaagaacgg gaagcagggg acggtgcgcg agcgcgagcc gtcgtggcgt 120
gagcgggaga acaacaacca gcgccgggag cgccgtcgtc gggtggtcgc gtcgaggatc 180
ttcgccgggc tgcgcagata cggcaactac gccctcccca ggcactacga caacaacgtc 240
gtgctcatgg cgctctgcga ggaggccggc tggaccgtgg aggccgacgg caccacctac 300
cgcaggggag gaaagcctcc ggcaggtgat cagcatatgg ccgacattgg cgggtctgct 360
gctccggtga cccaccaggg agcttccgac ggtgggggca gcgctagcgg cggagccgac 420
cccgttccgg cgtggctcaa gaacctgtcc aagcagctca gcgactcgtc gtaccccaac 480
tacttcgcga gctcctccag ctccaacgcg ccggcgacgc cgcacaacgg ctcgccatcc 540
tcgtccccgc ggcgcctccg caagatggcg cgctactcct ccccgccgtc caccccgccg 600
ccgagcccgg cgagggcctc cgacatgctg ccgccatggg ccatacgcga tggtggcggc 660
agccgctact ccttccaggc gttctcattc cacgggagcg gcgatgggga gcaggcgggc 720
gcgaaggagg gggagggaaa aggaggagga gaaggtgcaa cgccacgagg aagaggttgc 780
agccacatat gccggggtgc aagccatgag gaggaggtct cgagcataga tgatggggtg 840
caggaccatg agaaggtcca ggcggatgac gagaccatcg aggctcctgc ctttcgacgg 900
aggggcactc ggaagggcca ctttgtcctt cctttgtcag agcctgcaca atcagatgct 960
cggagacgct atcgtgtgga acctgaggct caggagcatg ccgaccaagt attagagaaa 1020
aacgtgaaga aggttgtgaa tgtgtacatg aagcgtcgag gatattctat aaataacttt 1080
cggcaatatt cagatgtcta cttgaatgtg gaccagtaca tagagataca ggagtcttac 1140
aggcaggaga tagttaggaa atattctatg tttatcggtg tgatcgattc gacggagtta 1200
cggtctcgtg ggctgcttct tcgttgcagg caggttatgg tagcagttgt tagtcggggg 1260
ctccgagcat gggaggagtc tatgagggag cagcaagaaa ggttttcgtg a 1311
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence
<400> 2
<210> 3
<211> 22
<212> DNA
<213> Artificial sequence
<400> 3
tcacgaaaac ctttcttgct gc 22
<210> 4
<211> 44
<212> DNA
<213> Artificial sequence
<400> 4
tcctctagag tcgacatgga gggaggggta ggagcgggag gaag 44
<210> 5
<211> 47
<212> DNA
<213> Artificial sequence
<400> 5
caccatggta ctagtgtcga ccgaaaacct ttcttgctgc tccctca 47
<210> 6
<211> 41
<212> DNA
<213> Artificial sequence
<400> 6
tggagaggac agcccaagct tatggaggga ggggtaggag c 41
<210> 7
<211> 43
<212> DNA
<213> Artificial sequence
<400> 7
ctcaccatga ccggtggatc cgaaaacctt tcttgctgct ccc 43
<210> 8
<211> 20
<212> DNA
<213> Artificial sequence
<400> 8
<210> 9
<211> 22
<212> DNA
<213> Artificial sequence
<400> 9
tcacgaaaac ctttcttgct gc 22
<210> 10
<211> 2840
<212> DNA
<213> Artificial sequence
<400> 10
aattcccatg gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg 60
cgaacagttc atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat 120
ggtggagcac gacacgcttg tctactccaa aaatatcaaa gatacagtct cagaagacca 180
aagggcaatt gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg 240
cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg 300
ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa 360
agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc 420
aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta 480
tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga ggacagggta 540
cccggggatc ctctagagtc gacatggagg gaggggtagg agcgggagga agtcggggtg 600
atgagggttg ggctagggca ggagacgggg acgggaagaa cgggaagcag gggacggtgc 660
gcgagcgcga gccgtcgtgg cgtgagcggg agaacaacaa ccagcgccgg gagcgccgtc 720
gtcgggtggt cgcgtcgagg atcttcgccg ggctgcgcag atacggcaac tacgccctcc 780
ccaggcacta cgacaacaac gtcgtgctca tggcgctctg cgaggaggcc ggctggaccg 840
tggaggccga cggcaccacc taccgcaggg gaggaaagcc tccggcaggt gatcagcata 900
tggccgacat tggcgggtct gctgctccgg tgacccacca gggagcttcc gacggtgggg 960
gcagcgctag cggcggagcc gaccccgttc cggcgtggct caagaacctg tccaagcagc 1020
tcagcgactc gtcgtacccc aactacttcg cgagctcctc cagctccaac gcgccggcga 1080
cgccgcacaa cggctcgcca tcctcgtccc cgcggcgcct ccgcaagatg gcgcgctact 1140
cctccccgcc gtccaccccg ccgccgagcc cggcgagggc ctccgacatg ctgccgccat 1200
gggccatacg cgatggtggc ggcagccgct actccttcca ggcgttctca ttccacggga 1260
gcggcgatgg ggagcaggcg ggcgcgaagg agggggaggg aaaaggagga ggagaaggtg 1320
caacgccacg aggaagaggt tgcagccaca tatgccgggg tgcaagccat gaggaggagg 1380
tctcgagcat agatgatggg gtgcaggacc atgagaaggt ccaggcggat gacgagacca 1440
tcgaggctcc tgcctttcga cggaggggca ctcggaaggg ccactttgtc cttcctttgt 1500
cagagcctgc acaatcagat gctcggagac gctatcgtgt ggaacctgag gctcaggagc 1560
atgccgacca agtattagag aaaaacgtga agaaggttgt gaatgtgtac atgaagcgtc 1620
gaggatattc tataaataac tttcggcaat attcagatgt ctacttgaat gtggaccagt 1680
acatagagat acaggagtct tacaggcagg agatagttag gaaatattct atgtttatcg 1740
gtgtgatcga ttcgacggag ttacggtctc gtgggctgct tcttcgttgc aggcaggtta 1800
tggtagcagt tgttagtcgg gggctccgag catgggagga gtctatgagg gagcagcaag 1860
aaaggttttc gactagtacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 1920
ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 1980
gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 2040
tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 2100
gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 2160
tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 2220
agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 2280
acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 2340
tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 2400
acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 2460
tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 2520
agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 2580
tggacgagct gtacaagtaa ctgcaggcat gccagggctc tcaatggagt ttgaatcaaa 2640
tcttccagct gctttaatga gatatgcgag acgcctatga tcgcatgata tttgctttca 2700
attctgttgt gcacgttgta aaaaacctga gcatgtgtag ctcagatcct taccgccggt 2760
ttcggttcat tctaatgaat atatcacccg ttactatcgt atttttatga ataatattct 2820
ccgttcaatt tactgattga 2840
Claims (9)
1. The application of the corn ZmBES1/BZR1-3 gene in increasing the seed yield of plants is characterized in that the nucleotide sequence of the corn ZmBES1/BZR1-3 gene is shown as SED ID NO. 1.
2. The application of the corn ZmBES1/BZR1-3 gene in improving plant seed phenotype is characterized in that the nucleotide sequence of the corn ZmBES1/BZR1-3 gene is shown as SED ID NO. 1.
3. The method of any one of claims 1-2, wherein said plant is selected from the group consisting of arabidopsis thaliana and rice.
4. The use of claim 2, wherein said plant seed phenotype is seed length, seed width and thousand kernel weight.
5. A method for cultivating high-yield plants is characterized in that the expression of corn ZmBES1/BZR1-3 genes in the plants is up-regulated to obtain plants with over-expressed ZBES 1/BZR1-3 genes, thereby obtaining the high-yield plants.
6. The method of claim 5, comprising the steps of: separating corn ZmBES1/BZR1-3 genes from corn seedling leaves, designing amplification primers of the corn ZmBES1/BZR1-3 genes, amplifying cDNA sequences of the corn ZmBES1/BZR1-3 genes by a PCR method, constructing 35S-ZBES 1/BZR1-3-eGFP vectors by using the cDNA sequences of the ZmBES1/BZR1-3 genes, transforming the vectors into target plants by using an agrobacterium-mediated method, improving the expression of the ZmBES1/BZR1-3 genes, and obtaining over-expressed ZBES 1/BZR1-3 genes, thereby obtaining high-yield plants.
7. The method of any one of claims 5 to 6, wherein the maize ZmBES1/BZR1-3 gene has the nucleotide sequence set forth in SED ID No. 1.
8. The method as claimed in any one of claims 5 to 6, wherein the nucleotide sequence of the amplification primer of the maize ZmBES1/BZR1-3 gene is as defined in SED ID No.2 and SED ID No. 3.
9. The method according to any one of claims 5 to 6, wherein the plant is one of Arabidopsis thaliana or rice.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210270506.9A CN114672511B (en) | 2022-03-18 | 2022-03-18 | Application of corn ZmBES1/BZR1-3 gene in increasing plant seed yield |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210270506.9A CN114672511B (en) | 2022-03-18 | 2022-03-18 | Application of corn ZmBES1/BZR1-3 gene in increasing plant seed yield |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114672511A true CN114672511A (en) | 2022-06-28 |
| CN114672511B CN114672511B (en) | 2023-04-25 |
Family
ID=82074393
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210270506.9A Active CN114672511B (en) | 2022-03-18 | 2022-03-18 | Application of corn ZmBES1/BZR1-3 gene in increasing plant seed yield |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114672511B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115851753A (en) * | 2022-07-06 | 2023-03-28 | 四川农业大学 | Application of corn ZmBES1/BZR1-1 gene in improving plant yield |
| CN116732088A (en) * | 2023-08-10 | 2023-09-12 | 浙江大学海南研究院 | Application of PpyBZR2 gene in promoting pear dormancy bud germination |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090013433A1 (en) * | 2007-01-10 | 2009-01-08 | The Salk Institute For Biological Studies | Compositions, cells, and plants that include BKI1, a negative regulator of BRI1-mediated BR signaling |
| US20120266327A1 (en) * | 2009-10-22 | 2012-10-18 | Crop Functional Genomics Center | Plants Having Enhanced Yield-Related Traits and a Method for Making the Same |
| CN105087633A (en) * | 2014-04-29 | 2015-11-25 | 中国科学院上海生命科学研究院 | Gene for regulating plant height, tiller number and leaf inclination and application thereof |
| CN105420245A (en) * | 2015-11-19 | 2016-03-23 | 中国农业科学院生物技术研究所 | Salt-stress induced gene OsEAR1 of plants and encoding protein and application thereof |
| US20180094275A1 (en) * | 2014-11-04 | 2018-04-05 | Agresearch Limited | Methods for Plant Improvement |
| CN111440804A (en) * | 2020-04-30 | 2020-07-24 | 四川农业大学 | Application of corn ZmBES1/BZR1-5 gene in cultivation of large-grain plants |
| CN112119163A (en) * | 2018-02-16 | 2020-12-22 | 首尔大学校产学协力团 | Transgenic plants with increased yield |
| CN112226455A (en) * | 2019-06-27 | 2021-01-15 | 中国科学院植物研究所 | Rice grain length and grain weight related protein, and coding gene and application thereof |
| CN112941087A (en) * | 2021-04-06 | 2021-06-11 | 四川农业大学 | Application of corn ZmBES1/BZR1-2 gene in improving plant drought tolerance |
| CN113121662A (en) * | 2019-12-31 | 2021-07-16 | 中国科学院植物研究所 | Cotton GhBZR3 protein and application of coding gene thereof in regulating plant growth and development |
| CN114480431A (en) * | 2022-03-30 | 2022-05-13 | 四川农业大学 | Application of corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants |
| CN114480422A (en) * | 2022-02-09 | 2022-05-13 | 四川农业大学 | Application of corn ZmBES1/BZR1-9 gene in breeding early flowering plants |
-
2022
- 2022-03-18 CN CN202210270506.9A patent/CN114672511B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090013433A1 (en) * | 2007-01-10 | 2009-01-08 | The Salk Institute For Biological Studies | Compositions, cells, and plants that include BKI1, a negative regulator of BRI1-mediated BR signaling |
| US20120266327A1 (en) * | 2009-10-22 | 2012-10-18 | Crop Functional Genomics Center | Plants Having Enhanced Yield-Related Traits and a Method for Making the Same |
| CN105087633A (en) * | 2014-04-29 | 2015-11-25 | 中国科学院上海生命科学研究院 | Gene for regulating plant height, tiller number and leaf inclination and application thereof |
| US20180094275A1 (en) * | 2014-11-04 | 2018-04-05 | Agresearch Limited | Methods for Plant Improvement |
| CN105420245A (en) * | 2015-11-19 | 2016-03-23 | 中国农业科学院生物技术研究所 | Salt-stress induced gene OsEAR1 of plants and encoding protein and application thereof |
| CN112119163A (en) * | 2018-02-16 | 2020-12-22 | 首尔大学校产学协力团 | Transgenic plants with increased yield |
| CN112226455A (en) * | 2019-06-27 | 2021-01-15 | 中国科学院植物研究所 | Rice grain length and grain weight related protein, and coding gene and application thereof |
| CN113121662A (en) * | 2019-12-31 | 2021-07-16 | 中国科学院植物研究所 | Cotton GhBZR3 protein and application of coding gene thereof in regulating plant growth and development |
| CN111440804A (en) * | 2020-04-30 | 2020-07-24 | 四川农业大学 | Application of corn ZmBES1/BZR1-5 gene in cultivation of large-grain plants |
| CN112941087A (en) * | 2021-04-06 | 2021-06-11 | 四川农业大学 | Application of corn ZmBES1/BZR1-2 gene in improving plant drought tolerance |
| CN114480422A (en) * | 2022-02-09 | 2022-05-13 | 四川农业大学 | Application of corn ZmBES1/BZR1-9 gene in breeding early flowering plants |
| CN114480431A (en) * | 2022-03-30 | 2022-05-13 | 四川农业大学 | Application of corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants |
Non-Patent Citations (5)
| Title |
|---|
| FUAI SUN等: "Maize transcription factor ZmBES1/BZR1-5 positively regulates kernel size" * |
| WENQI FENG等: "Maize ZmBES1/BZR1-3 and -9 Transcription Factors Negatively Regulate Drought Tolerance in Transgenic Arabidopsis" * |
| YU,H.等: "Zea mays BES1/BZR family protein BES1/BZR3 mRNA, complete cds" * |
| 冯文奇等: "玉米转录因子ZmBES1/BZR1-7基因克隆及功能分析" * |
| 明川: "玉米BES1/BZR1转录因子基因鉴定" * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115851753A (en) * | 2022-07-06 | 2023-03-28 | 四川农业大学 | Application of corn ZmBES1/BZR1-1 gene in improving plant yield |
| CN115851753B (en) * | 2022-07-06 | 2024-03-15 | 四川农业大学 | Application of corn ZmBES1/BZR1-1 gene in improving plant yield |
| CN116732088A (en) * | 2023-08-10 | 2023-09-12 | 浙江大学海南研究院 | Application of PpyBZR2 gene in promoting pear dormancy bud germination |
| CN116732088B (en) * | 2023-08-10 | 2023-11-07 | 浙江大学海南研究院 | Application of PpyBZR2 gene in promoting pear dormancy bud germination |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114672511B (en) | 2023-04-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114672511B (en) | Application of corn ZmBES1/BZR1-3 gene in increasing plant seed yield | |
| CN114480431B (en) | Application of corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants | |
| CN106868021B (en) | Gene OsNAC1 for controlling rice seed size and application thereof | |
| CN111440804B (en) | Application of corn ZmBES1/BZR1-5 gene in cultivation of large-grain plants | |
| CN106754957B (en) | OsSCAMP13 gene, encoding protein, application of stress resistance and acquisition method | |
| CN114480422B (en) | Application of corn ZmBES1/BZR1-9 gene in breeding early flowering plants | |
| CN115851823B (en) | Cymbidium CgARF18 gene and application thereof | |
| CN113584051B (en) | Application of GhGAI gene in regulation and control of plant flowering | |
| CN113337522B (en) | Application of cotton GhNFYC4 gene in promoting plant flowering | |
| CN110951771B (en) | Chinese cymbidiummiR390aApplication in controlling plant root system development | |
| CN110904106B (en) | Application of cymbidium goeringii miR159b in enhancing plant cold sensitivity | |
| CN108588069B (en) | Precursor gene of mulberry miR171a and application thereof in enhancing salt tolerance of plants | |
| CN108017697B (en) | Plant tillering angle related protein HSFA2D, and coding gene and application thereof | |
| CN112501181A (en) | Rice stress resistance related gene OsTZF7 and encoding protein and application thereof | |
| CN115851753B (en) | Application of corn ZmBES1/BZR1-1 gene in improving plant yield | |
| CN116254288B (en) | Application of cymbidium MIR156b gene in regulating and controlling flowering time of plants | |
| CN116179590B (en) | Application of cymbidium miR396 gene in regulation and control of thickening of plant stems | |
| CN116042696B (en) | Application of cymbidium MIR156a gene in regulating and controlling plant fruit development | |
| CN111304198B (en) | Application of cymbidium goeringii miR390b in controlling plant vegetative organ development | |
| CN111607604B (en) | Application of cotton GHPSAT2 gene in promoting flowering of plants | |
| CN117660485A (en) | Application of Arabidopsis ERF012 gene in regulation of seed germination | |
| CN117511994A (en) | Application of corn ZmBES1/BZR1-4 gene in increasing seed yield | |
| CN116640769A (en) | Peanut AhGATA11 gene and application thereof in improving stress resistance of plants | |
| CN117230083A (en) | Application of over-expression GhVOZ1 gene in promotion of cotton flowering | |
| CN115992150A (en) | Application of GhbHLH093 gene in regulation of flowering phase of plants |
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 |