CN110042108A - A kind of plant type of rice growth and development correlative coding gene and its application - Google Patents
A kind of plant type of rice growth and development correlative coding gene and its application Download PDFInfo
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- CN110042108A CN110042108A CN201910251175.2A CN201910251175A CN110042108A CN 110042108 A CN110042108 A CN 110042108A CN 201910251175 A CN201910251175 A CN 201910251175A CN 110042108 A CN110042108 A CN 110042108A
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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
Abstract
The invention discloses a kind of plant type of rice growth and development correlative coding genes, the plant type of rice growth and development correlative coding Gene Name is OrMKK3-2, application of the plant type of rice growth and development correlative coding gene OrMKK3-2 in controlling plant type of rice growth and development.First identified of the present invention and plant type of rice growth and development related gene OrMKK3-2, plant type of rice growth and development related gene OrMKK3-2 can cause plant type of rice to be downgraded under conditions of function enhances or expression quantity rises, tiller increases proves that plant type of rice growth and development related gene albumen or its albumen play a significant role in control plant type of rice.
Description
Technical field
The present invention relates to gene engineering technology field, in particular to a kind of plant type of rice growth and development correlative coding gene and
It is applied.
Background technique
Rice is one of most important cereal crops of the mankind, accounts for about 50% by the population of staple food of rice in the world.Asia
Area increases the demand of rice year by year, since rapid economic development, population pressure increase, industrial land increases severely, arable land face
Product is reduced, and Rice Production faces huge pressure.Therefore high yield potential that is constantly improve and excavating rice, is the master of scientists
Want research direction.Plant type of rice and quality, yield have close relationship.Plant type of rice is in group's light posture, luminous energy, water
Key effect is played on fertilizer, space utilization rate, output increased and quality-improving.Plant type depends primarily on plant type, blade profile, tiller number
Several aspects such as mesh, tillering angle and fringe portion form.In recent years, become using molecular biology method cloning rice plant type gene
The rice plant type gene of the hot spot of research, clone has important theory and practical guidance to rice dwarf breeding, plant type improvement
Meaning.
The rice plant type gene for having cloned clear function at present reaches 21, sd1, d2 including No. 1 chromosome of rice,
The genes such as d10, d18, d61, gid2, gid1 control semi-dwarf mutant and short bar character, and the gid2 gene of No. 2 chromosome controls short bar
Character, No. 3 chromosome d14, brd1, fc1, OsDWARF4 gene control plant types are formed, and the d11 gene of No. 4 chromosome controls water
The short bar of the more tillers of rice, the eui1 control internode beneath spike elongation of No. 5 chromosome, No. 6 chromosomes d3 and d35 control short bar character, No. 7 dyes
Colour solid d6 gene, No. 8 chromosome SPY, IPA1 genes, the brd2 gene of No. 10 chromosome control the formation of plant type of rice.So
And the genetic entities comprehensive agronomy trait expression for the control plant type of rice character excavated and identified at present is poor, current plant type is educated
The main problem of kind is that applicable plant type gene is very little, and how breakthrough bottleneck applies new plant type gene to seem particularly critical.
Be richly stored with gene in oryza officinalis, excavates the beneficial gene of oryza officinalis.It is excavated from wild rice and identifies strain
Type new resources deeply probe into the molecular mechanism of regulation plant type, it will help rice breeding man is MOLECULE DESIGN in breeding process
Breeding improvement rice plant and leaf shape provides theoretical foundation, finally realizes high-efficient breeding.
The information disclosed in the background technology section is intended only to increase the understanding to general background of the invention, without answering
When being considered as recognizing or imply that the information constitutes the prior art already known to those of ordinary skill in the art in any form.
Summary of the invention
The present invention in view of the above technical problems, invents a kind of plant type of rice growth and development correlative coding gene and its application.
To achieve the above object, technical solution provided by the invention is as follows:
A kind of plant type of rice growth and development correlative coding gene, entitled OrMKK3-2 derive from Oryza oryza officinalis
The cDNA sequence of the Y003 strain of (Oryza officinalis Wall.ex wstt), 4 kinds for belonging to the same gene are variable
Shear second of open reading frame of open reading frame.
Wherein, the plant type of rice growth and development correlative coding gene OrMKK3-2 is a) or b):
A) cDNA sequence gene order as shown in SEQ ID No.1, by 1556 base compositions；
B) substitution and/or missing by gene order shown in SEQ ID No.1 Jing Guo one or several bases and/or add
The protein as derived from a) of the OrMKK3-2 protein active related gene coded with plant type of rice added.
A kind of plant type of rice growth and development correlative coding gene, entitled OrMKK3-2, in controlling plant type of rice and tiller
Developmental application.
Wherein, the controlling plant type of rice growth and development refers to adjusting and controlling rice dwarfing, more tillers.
In order to solve the above technical problems, the present invention also provides the plant type of rice growth and development related gene OrMKK3-2
Application in the transgenic paddy rice for cultivating of short stem, more tiller rice.
Wherein, biomaterial relevant to the plant type of rice growth and development related gene OrMKK3-2, cultivate it is of short stem,
Application in the transgenic paddy rice of more tiller rice；The life relevant to plant type of rice growth and development related gene OrMKK3-2
Object material is following A 1) any one of to A20):
A1) nucleic acid molecules 1；The nucleic acid molecules 1 are to encode the plant type of rice growth and development related gene OrMKK3-2
Nucleic acid molecules；
A2) contain A1) expression cassettes of the nucleic acid molecules 1；
A3) contain A1) recombinant vectors of the nucleic acid molecules 1；
A4) contain A2) recombinant vector of the expression cassette；
A5) contain A1) recombinant microorganisms of the nucleic acid molecules 1；
A6) contain A2) recombinant microorganism of the expression cassette；
A7) contain A3) recombinant microorganism of the recombinant vector；
A8) contain A4) recombinant microorganism of the recombinant vector；
A9) contain A1) the transgenic plant cells systems of the nucleic acid molecules 1；
A10) contain A2) the transgenic plant cells system of the expression cassette；
A11) contain A3) the transgenic plant cells system of the recombinant vector；
A12) contain A4) the transgenic plant cells system of the recombinant vector；
A13) contain A1) Transgenic plant tissues of the nucleic acid molecules 1；
A14) contain A2) Transgenic plant tissue of the expression cassette；
A15) contain A3) Transgenic plant tissue of the recombinant vector；
A16) contain A4) Transgenic plant tissue of the recombinant vector；
A17) contain A1) the genetically modified plants organs of the nucleic acid molecules 1；
A18) contain A2) the genetically modified plants organ of the expression cassette；
A19) contain A3) the genetically modified plants organ of the recombinant vector；
A20) contain A4) the genetically modified plants organ of the recombinant vector.
Above-mentioned biomaterial relevant to the plant type of rice growth and development related gene OrMKK3-2 is in adjusting and controlling rice strain
In application in type growth and development or the application in the transgenic paddy rice for cultivating Semidwarf rice, the nucleic acid molecules be can be
DNA, such as cDNA, genomic DNA or recombinant DNA；The nucleic acid molecules are also possible to RNA, such as mRNA or hnRNA.
In above-mentioned biomaterial relevant to the plant type of rice growth and development related gene OrMKK3-2, the expression cassette
It is the DNA for referring to express corresponding protein in host cell, which not only may include the starting for starting related gene transcription
Son may also include the terminator for terminating related gene transcription, such as A2) as described in containing encoding the plant type of rice growth and development phase
The expression cassette for closing the nucleic acid molecules of albumen OrMKK3-2, is to refer to express the plant type of rice growth and development in host cell
The DNA of related gene OrMKK3-2.
The recombinant vector of the OrMKK3-2 expression casette, such as pET- can be contained with existing plant expression vector construction
28a、pCAMBIA2301、pSP72、pROKII、pBin438、pCAMBIA1302、pCAMBIA1301、pCAMBIA1300、
PBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA company) etc..
In above-mentioned biomaterial, A5)-A8) in any recombinant microorganism or B5)-B8) and in any micro- life of recombination
Object concretely bacterium, yeast, algae or fungi.Wherein, bacterium may be from Escherichia (Escherichia), Erwinia
(Erwinia), Agrobacterium tumefaciems category (Agrobacterium), Flavobacterium (Flavobacterium), Alcaligenes
(Alcaligenes), pseudomonas (Pseudomonas), Bacillus (Bacillus) etc..A9)-A12) in any institute
The transgenic cell line stated, A13)-A16) in any Transgenic plant tissue, A17)-A20) in any described turn
Gene plant organ does not include the propagation material of plant.
It is provided by the present invention it is a kind of cultivate rice dwarf, more tillers transgenic paddy rice method, by SEQ ID No.1
Shown in plant type of rice growth and development correlative coding gene OrMKK3-2 building over-express vector Introduced into Rice in, obtain
The transgenic paddy rice that OrMKK3-2 gene expression dose improves: being to be overexpressed plant type of rice growth and development phase described in receptor rice
The expression of the encoding gene of correlation gene OrMKK3-2, the transgenic paddy rice downgraded.
In the above method, the expression of the encoding gene of the plant type of rice growth and development related gene OrMKK3-2 is to pass through
It will be as realized in the DNA fragmentation of SEQ ID No.1 importing receptor rice.
In the above method, the expression of the encoding gene of the plant type of rice growth and development related gene OrMKK3-2 can pass through
Over-express vector, which is imported into, to be realized in receptor rice.
Wherein, the over-express vector is recombinant expression carrier PMDC32 or carrier pCAMBIA1301；The recombinant expression
Carrier PMDC32 is that the sequence between Asc I and the PacI recognition site by expression vector PMDC32 or carrier pCAMBIA1301 is replaced
It is changed to DNA sequence dna shown in SEQ ID No.1.
Wherein, the recombinant expression carrier PMDC32 can by using mediated by agriculture bacillus, Ti-plasmids, plant viral vector,
Directly delivered DNA, microinjection, the standard biologics technical method such as electroporation import plant cell or tissue.
Wherein, the method also includes described in the screening from the receptor rice for importing DNA molecular shown in SEQ ID No.1
The raised rice of plant type of rice growth and development related gene OrMKK3-2 expression quantity, obtains the OrMKK3-2 gene expression dose
The step of raised transgenic paddy rice.
Wherein, the transgenic paddy rice is interpreted as the not only first generation turn comprising obtaining the genetic transformation receptor rice
Trans-genetic hybrid rice also includes its filial generation.For transgenic paddy rice, the gene can be bred in the species, it is also possible to conventional breeding
The gene transfer is entered other kinds of same species by technology, particularly including in commercial variety.The transgenic paddy rice includes
Seed, callus, intact plant and cell.
Above, the rice concretely OryzasativaLcv.Nipponbare.
Compared with prior art, the invention has the following beneficial effects:
First identified of the present invention and plant type of rice growth and development related gene OrMKK3-2, plant type of rice growth and development phase
Correlation gene OrMKK3-2 can cause plant type of rice dwarfing, tiller to increase proof water under conditions of function enhances or expression quantity rises
Rice plant type growth and development related gene albumen or its albumen play a significant role in control plant type of rice.The present invention be only into
The molecule mechanism that one step illustrates plant type of rice provides basis, and new genetic resources and breeding resources are provided for rice breeding.
The raised transgenic paddy rice of OrMKK3-2 gene expression that the present invention obtains can be used for studying as new rice germplasm material
The gene of the more controlling plant type of rice developments of the molecule mechanism and discovery that rice dwarf, tiller increase.The present invention is for passing through
Effectively controlling plant type of rice has important application value using the genetic resources for genetic breeding and gene engineering method.
Detailed description of the invention
Fig. 1 is the transcription water of the raised OrMKK3-2 gene for being overexpressed transgenic paddy rice of OrMKK3-2 gene expression dose
Flat detection；Wherein, WT represents receptor parent OryzasativaLcv.Nipponbare plant, and OE-OrMKK3-2-1 representative is transferred to recombinant vector PMDC32-
The overexpression positive plant of OrMKK3-2；OE-OrMKK3-2 plant, OE-OrMKK3-2-2 representative are transferred to recombinant vector PMDC32-
The overexpression positive plant OE-OrMKK3-2 plant of OrMKK3-2, OE-OrMKK3-2-1 are different from OE-OrMKK3-2-2
Transformation event.
Fig. 2 is the raised Phenotypic Observation for being overexpressed transgenic paddy rice of OrMKK3-2 gene expression dose；Wherein, WT is represented
Receptor parent OryzasativaLcv.Nipponbare plant, OE-OrMKK3-2-1 represent the overexpression positive plant for being transferred to recombinant vector PMDC32-OrMKK3-2
Strain OE-OrMKK3-2 plant, OE-OrMKK3-2-2 represent the overexpression positive plant for being transferred to recombinant vector PMDC32-OrMKK3-2
Strain OE-OrMKK3-2 plant, OE-OrMKK3-2-1 are different transformation events from OE-OrMKK3-2-2.
Specific embodiment
Specific embodiment is described in detail with reference to the accompanying drawing, it is to be understood that protection scope of the present invention not by
The limitation of specific embodiment.
Experimental method in following embodiments is unless otherwise specified conventional method.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Rice OryzasativaLcv.Nipponbare (also referred to as wild rice, abbreviation WT) in following embodiments is commercially available gained, and the public can also
The biomaterial is obtained from Guangxi Autonomous Region Academy of Agricultural Sciences's rice research, which only attaches most importance to what duplicate was invented
Used in related experiment, it not can be used as other purposes and use.
Expression carrier used thereof PMDC32 is commercially available gained in following embodiments, and the public can also therefrom Guangxi Zhuang Autonomous Region agriculture
Industry academy of sciences rice research obtains the biomaterial, which only attaches most importance to used in the related experiment of duplicate invention, can not
It is used as other purposes.
Agrobacterium in following embodiments is Agrobacterium tumefaciems EHA105 (Agrobacterium tumefaciens
EHA105)(New Agrobacterium helper plasmids for gene transfer toplants.Hood,
ElizabethE；Gelvin,StantonB；Melchers,Leo S；Hoekema,Andre.Transgenic research,2
It (4): p.208-218 (1993) is), commercially available gained, the public can also be from rice research institute, Guangxi Autonomous Region Academy of Agricultural Sciences
The biomaterial is obtained, which only attaches most importance to used in the related experiment of duplicate invention, not can be used as other purposes and uses.
Embodiment 1
The building of plant type of rice growth and development related gene OrMKK3-2 gene overexpression carrier
1, the acquisition of plant type of rice growth and development related gene OrMKK3-2 gene
Using the reverse transcription cDNA of oryza officinalis Y003 (Oryza officinalis Wall.ex wstt) as template, use
Following primer primer1 and primer2 carry out PCR amplification and obtain target gene.
Primer1:5'TTGGCGCGCCCTGCAGGTTTAAACGAATTGCCCTT 3'；
Primer2:5'CCTTAATTAATGCTGCTCTCCCTTCCTTTTGTGCT 3'.
It will be connected after PCR product recovery purifying and (buy from Beijing Quan Shi King Company) sequencing vector into Zero, convert DH5 α
Competent cell after selecting positive colony, is sequenced.
Sequencing result shows that the PCR product expanded has 4 variable sheers, and second is sequence such as SEQ ID No.1
Shown in nucleotide sequence, length 1556bp is named as OrMKK3-2 gene.The ammonia of the protein of OrMKK3-2 gene coding
Base acid sequence is as shown in SEQ ID No.2.
2, plant type of rice growth and development related gene OrMKK3-2 gene overexpression carrier (recombinant expression carrier PMDC32-
OrMKK3-2 building)
1) oryza officinalis cDNA is expanded with primer1 and primer2, obtains the variable sheer of OrMKK3-2 gene, and
It is connected to recombinant vector Zero-OrMKK3-2, has obtained the positive colony of Zero-OrMKK3-2.With restriction enzyme A sc I
OrMKK3-2 segment is obtained with PacI digestion recombinant vector Zero-OrMKK3-2；
2) restriction enzyme A sc I and PacI digestion expression vector PMDC32 is used, linear expression vector PMDC32 is obtained,
Recycle the linear fragment；Segment OrMKK3-2 obtained in step 1) is integrated into line using the method for homologous recombination directed cloning
Property expression vector PMDC32 on (specific method refers to PMDC32 specification), obtain homologous recombination product 1, then by homologous recombination
Product 1 is transferred to DH5 α competent cell, and 37 DEG C of overnight incubations obtain recombinant vector PMDC32-OrMKK3-2；
3) recombinant vector PMDC32-OrMKK3-2 obtained by step 2) is sequenced, the results showed that the recombinant vector
PMDC32-OrMKK3-2 is inserted as shown in SEQ ID No.1 in the Asc I restriction enzyme site forward direction of expression vector PMDC32
Nucleotide sequence, i.e., successfully by the DNA sequence dna between Asc I and the PacI recognition site of PMDC32 (identification sequence) replace with as
DNA sequence dna shown in SEQ ID No.1.
Embodiment 2
Cultivate the raised overexpression OrMKK3-2 of plant type of rice growth and development related gene OrMKK3-2 gene expression dose
The identification of transgenic plant and transgenic plant
One, the raised overexpression OrMKK3-2 transgenic plant of OrMKK3-2 gene expression dose is cultivated by recombinant vector
PMDC32-OrMKK3-2 is by Agrobacterium tumefaciems EHA105 mediated transformation OryzasativaLcv.Nipponbare japonica rice, and the specific method is as follows:
1,1 gained recombinant vector PMDC32-OrMKK3-2 of embodiment heat shock method is imported in Agrobacterium tumefaciems EHA105
Obtain the recombination Agrobacterium tumefaciems EHA105 containing recombinant vector PMDC32-OrMKK3-2；Recombinant vector PMDC32- will be contained
The recombination Agrobacterium tumefaciems EHA105 of OrMKK3-2 collects thallus in 28 DEG C of culture 16h；It uses containing concentration as 100 μM of acetyl
Thallus is diluted by the N6 fluid nutrient medium (Sigma, catalog number C1416) of syringone, obtains dilution bacterium solution, dilution
The OD600 ≈ 0.5 of bacterium solution；
2, by culture to one month Mature Embryos of Rice embryo callus and the dilution bacterium solution mixed infection of step 1
30min is transferred in N6 solid co-cultivation medium after blotting bacterium solution using filter paper, in 24 DEG C of co-cultivation 3d, is obtained at co-cultivation
Callus after reason；
3, by step 2 to co-culture that treated callus is seeded in containing mass concentration be 150mg/L hygromycin
N6 solid screening and culturing medium (hygromycin is added into N6 solid medium and obtains N6 solid screening and culturing medium, the screening training of N6 solid
The mass concentration of hygromycin in base is supported to carry out first time screening on 150mg/L)；
4, being transferred in the 16th day picking health callus that first time screening starts containing mass concentration is 200mg/L tide
The N6 solid screening and culturing medium of mycin (is added hygromycin into N6 solid medium and obtains N6 solid screening and culturing medium, N6 solid
For the mass concentration of hygromycin to carry out programmed screening on 200mg/L), every 15 days subcultures are primary in screening and culturing medium, altogether subculture 1
It is secondary；
5, the resistant calli that picking step 4 obtains is transferred to trains containing the differentiation that mass concentration is 150mg/L hygromycin
(differential medium: 6-BA 2mg, NAA 0.2mg, N6 4g, caseinhydrolysate 1g, inositol 0.1g, sucrose 25g, sorb is supported on base
Alcohol 2.4g, agar powder 7g, deionized water 1L) broken up, in 24 DEG C of culture 45d, (above-ground plant parts height is about at this time
15cm), it opens bottleneck hardening 3 days, then transplants to greenhouse production, as turn PMDC32-OrMKK3-2 plant (T0 generation).For not
With transformation event be named as OE-OrMKK3-2-1 and OE-OrMKK3-2-2 is respectively represented and is transferred to recombinant vector PMDC32-
The overexpression positive plant of OrMKK3-2.
Two, the raised PMDC32-OrMKK3-2 of plant type of rice growth and development related gene OrMKK3-2 gene expression dose
The PCR of transgenic plant, which is identified, to be extracted
The T0 generation for turning PMDC32-OrMKK3-2 (OE-OrMKK3-2-1, OE-OrMKK3-2-2) plant that step 1 obtains
The genomic DNA of the seedling (referred to as WT) of seedling and receptor parent rice OryzasativaLcv.Nipponbare plant, and use primer primer1:5'
TTGGCGCGCCCTGCAGGTTTAAACGAATTGCCCTT 3' and primer2:5'CCTTAATTAATGCTGCTCTCCCTTCC
TTTTGTGCT 3' carries out PCR Molecular Detection and identifies positive seedling, obtains the plant of 1556bp PCR product as positive seedling, i.e., with
On OE-OrMKK3-2-1, OE-OrMKK3-2-2 for mentioning.
Three, the raised overexpression transgenic plant of plant type of rice growth and development related gene OrMKK3-2 gene expression dose
OrMKK3-2 gene expression dose identification:
OE-OrMKK3-2-1, OE-OrMKK3-2-2 plant and receptor parent rice Japan that extraction step two obtains respectively
The RNA of fine plant (referred to as WT) blade, sets internal reference as Actin, using internal control primer Actin-F and Actin-R, and
OrMKK3-2 gene specific, which quantifies primer OrMKK3-2-qRT-F and OrMKK3-2-qRT-R progress quantitative fluorescent PCR reaction, to be come
Detect the variation of the expression of different transgenic plant OrMKK3-2 genes.The result shows that (Fig. 1), is being transferred to recombinant vector
The expression of OrMKK3-2 gene is than the OrMKK3-2 gene of control (WT) in the positive plant of PMDC32-OrMKK3-2
Expression significantly rises, and above-mentioned primer is as follows:
Actin-F:5 '-ATTTGGCACCACACATTCTAC-3 '
Actin-R:5 '-ATAACCTTCGTAGATTGGGACT-3 '；
OrMKK3-2-qRT-F:5'CCCCGCCTACTTCTTCTTTC 3'
OrMKK3-2-qRT-R:5'CGCCGCCTTATCCATCTC 3'。
Four, the OrMKK3-2 that plant type of rice growth and development related gene OrMKK3-2 gene expression dose rises, which is overexpressed, turns
The phenotypic evaluation of gene plant
OE-OrMKK3-2-1, OE-OrMKK3-2-2 (ibid) plant that step 2 is obtained respectively and receptor parent rice
OryzasativaLcv.Nipponbare plant (referred to as WT) is planted in Hainan Experimental Base, observe in entire growth period PMDC32-OrMKK3-2 plant and
The phenotypic difference of receptor parent rice OryzasativaLcv.Nipponbare plant (referred to as WT).Measurement observation result such as Fig. 2, table 1, with receptor parent water
Rice OryzasativaLcv.Nipponbare plant is compared, and the phenotype of plant type dwarfing occurs in PMDC32-OrMKK3-2 plant, to demonstrate OrMKK3-2
Gene participates in the growth and development of control plant type of rice, i.e. the OrMKK3-2 gene is plant type of rice related gene.
1. plant type of rice growth and development related gene OrMKK3-2 of table is overexpressed transgenosis plant height and tiller shows
| Number | Plant height (cm) | Plant height standard error | Tiller (branch) | Tiller standard error |
| WT | 61.08 | 0.46 | 6.25 | 0.46 |
| OE-OrMKK3-2-1 | 38.67 | 0.24 | 9.75 | 0.72 |
| OE-OrMKK3-2-2 | 61.08 | 0.61 | 12.5 | 0.68 |
The aforementioned description to specific exemplary embodiment of the invention is in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to the above instruction, can much be changed
And variation.The purpose of selecting and describing the exemplary embodiment is that explaining specific principle of the invention and its actually answering
With so that those skilled in the art can be realized and utilize a variety of different exemplary implementation schemes of the invention and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Sequence table
<110>Guangxi Autonomous Region Academy of Agricultural Sciences's rice research institute
<120>a kind of plant type of rice growth and development correlative coding gene and its application
<130>Zhongyuweisheng Intellectual property rights agents Co., Ltd., Beijing city
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1556
<212> DNA
<213>a kind of plant type of rice growth and development correlative coding gene and its application (rice)
<400> 1
atggcggggc tcgaggagct gaagaagaag ctgcagcccc tgctgttcga cgacccggac 60
aagggcggcg tcagtagcag ggttcctcta ccggaggaca cctgcgactc ctacgtggtt 120
tctgatggtg gaactgtgaa tttgttgagt agatcattgg gtgagtataa catcaatgag 180
catggctttc ataaacgaag cactgggcca gaggagtcag attctggtga aaaggcatac 240
cgatgtgcct ctcatgatat gcacatattt ggccccatcg gtaatggtgc aagcagtgtt 300
gtgcagagag ctgtttttat accagttcat cgaattttgg ccttgaagaa gataaatata 360
tttgagaagg agaagagaca acaaattctg aatgagatga gaacattatg tgaagcatgt 420
tgttatattg gtttagttga attccagggt gcattctaca tgcctgattc tggacaaata 480
agcatcgccc ttgaatacat ggatggtggg tccttagctg atgttataaa gattaagaaa 540
tcaataccag aaccagttct tgcacatatg ctgcagaaag tattgcttgg tttgcgctac 600
ttgcatgaag taagacatct agtgcataga gatataaagc cagcgaattt actggtaaat 660
ctcaagggcg aggcaaagat aacagatttt ggagtgagtg ctggtttgga taatacgatg 720
gccatggcac agtcacatat atgtcacctg agagaattcg taacgagaat tactcttatg 780
ctgctgatat ttggagtctt ggactagcag tattggagtg tgctactggt aaatttccat 840
ataatgtgaa tgaaggccca gccaacctca tgctgcagat tctggatgat ccatcaccaa 900
caccaccaaa agatgcttat tcatccgagt ttagttcgtt catcaatgac tgcttgcaga 960
aagatgctga tgcaaggcct tcgtgtgagc agcttttgtc acacccattc atcaagaggt 1020
atgaaaacac taccatggac ttggtagctt acatcaaaag tgttgttgat ccaacagaaa 1080
gattaaagca aatagcagag atgcttgccg tacattacta cctcctcttt aacggcactg 1140
atgggatttg gcattatatg aagacattct acatggaaga atcaactttc agtttctcag 1200
ggaatgtgta tgttggccaa agtgacatat ttgatacttt atcaaatata agaaagaagt 1260
taacaggtga tcgtcctcga gagaaaattg ttcatgttgt tgagaagcta cattgtcgtg 1320
cacacggaga aacaggaata gctatacgtg tgtctggatc gttcattgtg ggaaaccaat 1380
ttctaatatg tggtgaaggg ttgcaagctg aaggaatgcc aagcttggag gaactctcca 1440
ttgatattcc aagcaagcgg gtaggtcagt tccgggagca atttatcatg gagccaggaa 1500
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Met Ala Gly Leu Glu Glu Leu Lys Lys Lys Leu Gln Pro Leu Leu Phe
1 5 10 15
Asp Asp Pro Asp Lys Gly Gly Val Ser Ser Arg Val Pro Leu Pro Glu
20 25 30
Asp Thr Cys Asp Ser Tyr Val Val Ser Asp Gly Gly Thr Val Asn Leu
35 40 45
Leu Ser Arg Ser Leu Gly Glu Tyr Asn Ile Asn Glu His Gly Phe His
50 55 60
Lys Arg Ser Thr Gly Pro Glu Glu Ser Asp Ser Gly Glu Lys Ala Tyr
65 70 75 80
Arg Cys Ala Ser His Asp Met His Ile Phe Gly Pro Ile Gly Asn Gly
85 90 95
Ala Ser Ser Val Val Gln Arg Ala Val Phe Ile Pro Val His Arg Ile
100 105 110
Leu Ala Leu Lys Lys Ile Asn Ile Phe Glu Lys Glu Lys Arg Gln Gln
115 120 125
Ile Leu Asn Glu Met Arg Thr Leu Cys Glu Ala Cys Cys Tyr Ile Gly
130 135 140
Leu Val Glu Phe Gln Gly Ala Phe Tyr Met Pro Asp Ser Gly Gln Ile
145 150 155 160
Ser Ile Ala Leu Glu Tyr Met Asp Gly Gly Ser Leu Ala Asp Val Ile
165 170 175
Lys Ile Lys Lys Ser Ile Pro Glu Pro Val Leu Ala His Met Leu Gln
180 185 190
Lys Val Leu Leu Gly Leu Arg Tyr Leu His Glu Val Arg His Leu Val
195 200 205
His Arg Asp Ile Lys Pro Ala Asn Leu Leu Val Asn Leu Lys Gly Glu
210 215 220
Ala Lys Ile Thr Asp Phe Gly Val Ser Ala Gly Leu Asp Asn Thr Met
225 230 235 240
Ala Met Ala Gln Ser His Ile Cys His Leu Arg Glu Phe Val Thr Arg
245 250 255
Ile Thr Leu Met Leu Leu Ile Phe Gly Val Leu Asp Gln Tyr Trp Ser
260 265 270
Val Leu Leu Val Asn Phe His Ile Met Met Lys Ala Gln Pro Thr Ser
275 280 285
Cys Cys Arg Phe Trp Met Ile His His Gln His His Gln Lys Met Leu
290 295 300
Ile His Pro Ser Leu Val Arg Ser Ser Met Thr Ala Cys Arg Lys Met
305 310 315 320
Leu Met Gln Gly Leu Arg Val Ser Ser Phe Cys His Thr His Ser Ser
325 330 335
Arg Gly Met Lys Thr Leu Pro Trp Thr Trp Leu Thr Ser Lys Val Leu
340 345 350
Leu Ile Gln Gln Lys Asp Ser Lys Gln Arg Cys Leu Pro Tyr Ile Thr
355 360 365
Thr Ser Ser Leu Thr Ala Leu Met Gly Phe Gly Ile Ile Arg His Ser
370 375 380
Thr Trp Lys Asn Gln Leu Ser Val Ser Gln Gly Met Cys Met Leu Ala
385 390 395 400
Lys Val Thr Tyr Leu Ile Leu Tyr Gln Ile Glu Arg Ser Gln Val Ile
405 410 415
Val Leu Glu Arg Lys Leu Phe Met Leu Leu Arg Ser Tyr Ile Val Val
420 425 430
His Thr Glu Lys Gln Glu Leu Tyr Val Cys Leu Asp Arg Ser Leu Trp
435 440 445
Glu Thr Asn Phe Tyr Val Val Lys Gly Cys Lys Leu Lys Glu Cys Gln
450 455 460
Ala Trp Arg Asn Ser Pro Leu Ile Phe Gln Ala Ser Gly Val Ser Ser
465 470 475 480
Gly Ser Asn Leu Ser Trp Ser Gln Glu Val Pro Trp Asp Ala Thr Thr
485 490 495
Tyr Gly Lys Ile Tyr Thr Ser Ser Lys Pro
500 505
Claims (9)
1. a kind of plant type of rice growth and development correlative coding gene, it is characterised in that: the plant type of rice growth and development correlation is compiled
Code Gene Name is OrMKK3-2；
Wherein, the plant type of rice growth and development correlative coding gene OrMKK3-2 is a) or b):
A) cDNA sequence gene order as shown in SEQ ID No.1, by 1556 base compositions；
B) it the substitution by gene order shown in SEQ ID No.1 Jing Guo one or several bases and/or is deleted and/or added
The protein as derived from a) of the OrMKK3-2 protein active related gene coded with plant type of rice arrived.
2. plant type of rice growth and development correlative coding gene according to claim 1, it is characterised in that: in controlling plant type of rice
And the application in tiller development.
3. plant type of rice growth and development correlative coding gene according to claim 2, it is characterised in that: the adjusting and controlling rice
Plant type growth and development refers to adjusting and controlling rice dwarfing, more tillers.
4. the plant type of rice growth and development correlative coding gene according to Claims 2 or 3, it is characterised in that: the rice strain
Application of the type growth and development related gene OrMKK3-2 in the transgenic paddy rice for cultivating of short stem, more tiller rice.
5. a kind of method for the transgenic paddy rice for being cultivated dwarfing, more tillers using gene as described in claim 1, feature are existed
In: the building over-express vector of plant type of rice growth and development correlative coding gene OrMKK3-2 shown in SEQ ID No.1 is imported
In rice, the transgenic paddy rice that OrMKK3-2 gene expression dose improves is obtained: being to be overexpressed rice strain described in receptor rice
The expression of the encoding gene of type growth and development related gene OrMKK3-2, the transgenic paddy rice downgraded.
6. according to the method described in claim 5, it is characterized by: the over-express vector be recombinant expression carrier PMDC32 or
Carrier pCAMBIA1301；The recombinant expression carrier PMDC32 is by expression vector PMDC32 or carrier pCAMBIA1301
Sequence between Asc I and PacI recognition site replaces with DNA sequence dna shown in SEQ ID No.1.
7. according to the method described in claim 5, it is characterized by: the recombinant expression carrier PMDC32 can be by using agriculture bar
Bacterium mediation, Ti-plasmids, plant viral vector, directly delivered DNA, microinjection, the standard biologics technical method such as electroporation, which imports, plants
Object cell or tissue.
8. according to the method described in claim 5, it is characterized by: the method also includes from shown in importing SEQ ID No.1
DNA molecular receptor rice in screen the raised water of the plant type of rice growth and development related gene OrMKK3-2 expression quantity
Rice, the step of obtaining the OrMKK3-2 gene expression dose raised transgenic paddy rice.
9. according to the method described in claim 5, it is characterized by: the rice concretely OryzasativaLcv.Nipponbare.
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| CN112626095A (en) * | 2020-12-21 | 2021-04-09 | 广西壮族自治区农业科学院 | Encoding gene related to common wild rice grain type and application thereof |
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