CN108866094A - Purposes of the rice uncertainty small ear gene OSIDS2 in adjusting and controlling rice yield - Google Patents
Purposes of the rice uncertainty small ear gene OSIDS2 in adjusting and controlling rice yield Download PDFInfo
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- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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- 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 belongs to technical field of molecular biology, especially purposes of the rice uncertainty small ear gene OSIDS2 in adjusting and controlling rice yield.The present invention also provides rice to spend more small ear gene OSIDS2 in the purposes improved in grass family cereal crops yield and the preparation for improving grass family cereal crops yield.The present invention provides a kind of new selection to improve grass family cereal crops yield.
Description
Technical field
The invention belongs to technical field of molecular biology, especially rice uncertainty small ear gene OSIDS2 in regulation water
Purposes in rice yield.
Background technique
In dicotyledon, terminal inflorescence separate living tissue is directly translated into floral meristem, and then is differentiated to form all kinds of
Floral organ.However, it is mitogenetic that terminal inflorescence separate living tissue is first converted into small ear in the monocotyledons gramineae plant such as rice
Tissue, is then transformed into floral meristem, then differentiate all kinds of floral organs again.And the development of small ear is a complexity and orderly
Spikelet development process is roughly divided into eight stages (Ikeda et al., 2004) according to the research of forefathers by process：
First stage, sterile glume former base are formed.Sterile glume former base is formed in a manner of 1/2 alternate phyllotaxy；
Second stage, the formation of sterile lemma former base.A pair of of sterile lemma former base is equally formed in a manner of 1/2 alternate phyllotaxy, secondary at this time
Sterile lemma stops development；
Phase III, lemma former base are formed.Lemma former base is formed in the top of external sterile lemma, at this time sterile lemma continued growth；
Fourth stage, glumelle former base are formed.Above internally positioned sterile lemma, with lemma positional symmetry, sterile lemma and lemma continue
Growth；
5th stage, lodicule former base are formed.Lemma side position formed two lodicule former bases, at this time sterile lemma, lemma,
Glumelle continued growth；
6th stage, stamen retrogressive are formed.Six pieces of stamen retrogressives are arranged in the form of verticil, and close to lemma side
It is slower to develop；
7th stage, carpellary primordia are formed.Floral meristem of the carpellary primordia in lemma side is formed, and stamen starts to break up
Transparent filigree and anther out；
8th stage, ovule and pollen formation phase.Floral meristem uncertainty is lost, and ovule former base is converted to.At this
Many important and complicated programs have occurred to complete the development of male and female reproductive organ during a.
Small ear is the distinctive inflorescence structure of all gramineous crops, the little Hua, little Hua comprising a pair of of bract and number not etc.
Number is usually 1-40, is the special inflorescence of one kind between inflorescence and flower.Small ear can be divided into two types：One is true
Qualitative small ear includes the little Hua to ascertain the number, such as rice, corn.Another kind is uncertain small ear, includes uncertain number
Little Hua, such as wheat.In certainty small ear, apical meristem is turned after the lateral meristem for generating fixed quantity
Turn to end floral meristem.And in uncertain small ear, side separate living tissue is constantly differentiated from apical meristem, top
Separate living tissue is not converted into end floral meristem.Currently, not finding to maintain small ear separate living tissue uncertainty destiny
Gene, on the contrary, have two genoids have the function of terminate small ear separate living tissue uncertainty destiny.The first kind can be defined as
Poppyhead separate living tissue characterizing gene.The LHS1 branch for having a grass family special in SEP family, in certainty small ear, LHS1
Genoid is only expressed in poppyhead, implies that it is probably gone the generation of specialization poppyhead separate living tissue by specifically requirement, thus
Realize small ear certainty (Cacharro ó n et al., 1999；Malcomber and Kellogg,2004；Zahn et
al.,2005).Second class is INDIERMINATE SPIKELET 1 (IDS1) genoid for belonging to AP2/ERF family, with LHS1
Genoid is different, does not determine poppyhead feature, and is responsible for starting on time small ear separate living tissue to floral meristem conversion.In jade
Meter Zhong, IDS1 gene mutation lead to produce additional little Hua (Chuck et al., 1998), the collateral line of IDS1 in a small ear
The single mutation of homologous gene SISTEROF IDS1 (SID1) does not have significant phenotype, but in ids1/sid1 double-mutant, it is small
Fringe develops multiple bract structures and cannot form little Hua (Chuck et al., 2008).In rice, two IDS1 genoids
The mutation of SUPERNUMERARYBRACT (SNB) and OsIDS1 results in sterile glume and increases, secondary in snb/osids1 double-mutant
Sterile lemma is more notable to be increased (Lee et al., 2006；Lee and An,2012).These are the result shows that IDS1 genoid is mutated
Extend small ear separate living tissue destiny.
Summary of the invention
The purpose of the present invention is to provide the nucleotide sequences and coding of rice uncertainty Spikelet development gene OSIDS2
Protein sequence provides powerful for Rice Panicle development research, promotes the raising of rice yield.
The present invention provides the preparation of adjusting and controlling rice yield, main active is the albumen of OSIDS2 gene coding
Matter, or the element or carrier of expression OSIDS2 gene.
Specifically, the protein that the OSIDS2 gene encodes has as SEQ ID No.1 or SEQ ID No.2 shows
Amino acid sequence.
Specifically, the OSIDS2 gene has such as SEQ ID No.3 or SEQ ID No.4 nucleotide sequence.
The present invention provides the preparation of adjusting and controlling rice grain number per spike, main active is the albumen of OSIDS2 gene coding
Matter, or the element or carrier of expression OSIDS2 gene.
Specifically, the protein that the OSIDS2 gene encodes has as SEQ ID No.1 or SEQ ID No.2 shows
Amino acid sequence.
Specifically, the OSIDS2 gene has such as SEQ ID No.3 or SEQ ID No.4 nucleotide sequence.
The present invention also provides purposes of the OSIDS2 gene in adjusting and controlling rice yield.
Specifically, the purposes is adjusting and controlling rice grain number per spike.
Specifically, the protein that the OSIDS2 gene encodes has as SEQ ID No.1 or SEQ ID No.2 shows
Amino acid sequence.
Specifically, the OSIDS2 gene has such as SEQ ID No.3 or SEQ ID No.4 nucleotide sequence.
Small ear is substantially a kind of inflorescence that grass family is special.A complete spike of rice is known as " fringe " in breeding, and this
Small ear in application refers to the small ear in auxology, the glumous flower being equivalent in thremmatology.Normal rice small ear is from base portion
It is successively made of to top a pair of of sterile glume, a pair of of sterile lemma and a basidixed little Hua.Basidixed little Hua includes 4 wheel floral organs, the 1st wheel
It is the lemma and glumelle locked together, lemma is slightly larger than glumelle；2nd wheel is raw two pieces of lodicules in lemma side；3rd wheel be
6 pieces of stamens, the 4th wheel is 1 piece of gynoecium, and gynoecium is located at the center of little Hua, stamen give birth to around it.Grain number per spike is the grass such as rice
The highly important yield traits of section cereal crops, constituent element include the number, length and small ear thereon that branch obstructs on inflorescence
Raw density etc., and the number of little Hua is not usually in the range of investigation in small ear, because small in one small ear of normal species
Flower number is stable.
Beneficial effects of the present invention：The present invention develops mutant oryza on the basis of the assignment of genes gene mapping, with Rice Panicle
Sativa indeterminate spikelet2 (osids2) is material, has cloned mutated gene OSIDS2.It is searched by homologous
Rope and gene order comparison in difference determine that OSIDS2 gene belongs to myb gene family MYB-1R class transcription factor.By aobvious outside 6
Son composition, gene cDNA overall length 921bp, encoder block DNA overall length 3887bp.In mutant osids2, cDNA is in the 5th exon
Upper 198th amino acids have the missing of a C base, cause frameshift mutation, and the 213rd amino acids is caused to be converted into termination codon
Son terminates protein translation in advance.Lead to part small ear glumelle lemma or degeneration, the small ear of part grows additional outer
Bran.Peel off flower glume and further look at discovery, inside the small ear of part have an additional glume shape organ, stamen number or increase
Or reduce, carpel number increases, and, the result shows that osids2 mutant small ear is intended to grow two little Hua, small ear is mitogenetic for these
Certainty is organized to lose.OSIDS2 gene maintains small ear to determine sexual development, and afunction leads to the increasing of little Hua number in small ear
Add, provides a kind of breeding methods that can increase grain number per spike.
Detailed description of the invention
Fig. 1 osids2 mutant and its small ear Phenotypic Observation at wild type heading stage
A1-A7, wild type (WT) small ear；A1-A3, stereoscope analysis；A4-A5, electron microscope analysis；A6-A7, transverse section tissue
Credit analysis；B1-B7, the osids2 mutant small ear of glumelle lemma；B1-B3, stereoscope analysis；B4-B5, electron microscope analysis；B6-
B7, transverse section histologic analysis；C1-C8, the osids2 mutant small ear that glumelle is degenerated；C1-C4, stereoscope analysis；C5-C6,
Electron microscope analysis；C7-C8, transverse section histologic analysis；Scale=250 μm.
The small ear morphologic observation of Fig. 2 osids2 mutant and its wild type panicle primordium dif ferentiation stage
A-D, wild type small ear；E-H, the osids2 that glumelle is degenerated；I-L, the osids2 small ear of glumelle lemma；Asterisk table
Show stamen.Scale=100 μm.
The map based cloning of Fig. 3 OSIDS2 gene
The finely positioning of A, OSIDS2 gene；B, OSIDS2 gene complementation carrier schematic diagram；C, OSIDS2 gene are wild
Real-time quantitative expression analysis in type and mutant children's fringe, ordinate indicate gene relative expression quantity；D, OSIDS2 gene complementation
Transgenic plant Phenotypic Observation scale=250 μm.
Fig. 4 OSIDS2 Gene Expression Profile Analysis
A, OSIDS2 gene are in Seedling Stage root, leaf, tillering stage root, leaf, bud, heading stage root, stem, section, sheath, leaf, auricle, bud
And the real-time quantitative expression point in different development stage (being less than 5cm, 0.5-2cm, 2-5cm) wild type small ear and each wheel floral organ
Analysis；B-D, OsMADS1, DL, OsMADS6 gene exist<5cm, 0.5-2cm, 2-5cm developmental stage wild type small ear and mutant are small
Real-time quantitative expression analysis in fringe, ordinate indicate gene relative expression quantity.
The amino acid alignment of Fig. 5 OSIDS2 albumen is analyzed
The subcellular localization of Fig. 6 OSIDS2 albumen
A-D:Negative control (35S-GFP)；E-H:The positioning (35S-OEOSIDS2-GFP) of OSIDS2 albumen
Specific embodiment
Material used in the embodiment of the present invention：Wild rice material red silk extensive No. 10 (WT) and uncertain small ear mutation
Body (osids2) is purchased from rice research institute of Southwest University； Reagent Kit With gDNA Eraser
It is Reverse Transcriptase kit, high-fidelity DNA polymerase Primer Star Max, Taq archaeal dna polymerase, T4DNA ligase, restricted
Restriction endonuclease, pMD19-T carrier, SYBR Green fluorescent dye and DL5,000 DNA Marker are purchased from TaKaRa company；；Ammonia benzyl
Penicillin (Ampicillin, Amp) and kalamycin (Kanamycin, Kan) are Sigma Products；DNA Marker
III, Ago-Gel DNA QIAquick Gel Extraction Kit, the small extraction reagent kit of rapid plasmid, plant total RNA extraction reagent box are purchased from Tiangeng
(TIANGEN) biotech firm, pEASY-Uni Seamless Cloning and Assembly Kit recombinase is purchased from full formula gold
Biotech firm, primer synthesis and DNA sequencing are completed by the handsome Bioisystech Co., Ltd in Shanghai；Other chemical reagent are purchased from Beijing
Ding Guo biotechnology Co., Ltd；PCAMBIA1300, pTCK303 plant expression vector, pA7 plant expression vector, large intestine
Bacillus DH5 α, Agrobacterium LBA4404 are bought in BioVector plasmid vector bacterium cell gene collection.
Used primer in 1 embodiment of table
The acquisition of 1 osids2 mutant of embodiment and phenotypic analysis
Rice flower organ mutant OSIDS2 is the natural mutation found in the breeding intermediate materials of Lu extensive 1089, warp
Cross inbreeding of more generation, inheritance stability.In research, with the Elite Rice restorer red silk extensive 10 of rice research institute of Chongqing Southwest University breeding
Number (J10) is used as wild type control, carries out correlation analysis to mutant.
In rice ear sprouting period, osids2 and the small ear of wild type is taken to carry out morphology and histologic analysis.Morphological analysis：
On the one hand it takes fresh mature small ear to be placed under Nikon SM1500 stereomicroscope dissected, observe flower organ morphology sum number
Mesh, and photograph to record；On the other hand take immature inflorescence and mature small ear using Hitachi SU3500 electronic scanner microscope at -20 DEG C
The observation of Electronic Speculum is scanned under vacuum condition.Histologic analysis：Take fresh small ear, FAA fixer (50% ethyl alcohol,
0.9M glacial acetic acid and 3.7% formaldehyde) in, overnight, by Gradient elution using ethanol, dimethylbenzene is replaced, finally by material packet for 4 DEG C of fixations
It is embedded in paraffin (Sigma).Material after embedding is cut into 8 μm of slice, then the material cut is placed in and fills poly
It on the glass slide of lysine, is dewaxed with dimethylbenzene, is dehydrated using the ethanol gradient of various concentration.With 1% sarranine and 1%
It is fast green that slice is dyed, it is then dehydrated by a series of ethanol gradients, is infiltrated and be sliced with dimethylbenzene, last natural gum envelope
Piece.Histologic analysis is carried out using Nikon E600 optical microscopy.As a result as follows：
In wild type, a normal rice small ear from top to bottom successively can including a pair of of sterile glume, a pair of of sterile lemma and one
The basidixed little Hua educated.Basidixed little Hua is made of (figure lemma, glumelle, two pieces of lodicules, one piece of gynoecium and six pieces of stamens around gynoecium
1, A1-A3).There are three types in osids2 mutant small ear, a kind of small ear has the glumelle or additional lemma of lemma
(B1-B7), a kind of small ear glumelle is degenerated, and lemma is bent (C1-C8) to glumelle to a certain extent, and in addition a kind of small ear is without obvious different
Chang Bingneng is normally solid.It peels off flower glume and further looks at discovery, part little Hua is except there are also additional glume shape organs for flower glume
(B1-B2, B4, B6-B7, C5), stamen number occur increasing or reducing (B2-B3, C2, C5), and observe that carpel increased shows
As (B6-B7), the formation (B6) for spending more small ear is implied.
In Sp4-Sp8 phase (the i.e. Spikelet development period 4-8, with reference to Itoh etc. 2005 of rice small ear early development differentiation
Document), take fresh wild type and osids2 mutant small ear, using Hitachi's SU3500 electronic scanner microscope -20 DEG C see
It examines.It in wild type small ear, has been formed in Sp4 period sterile glume and sterile lemma, while the inside and outside bran former base of poppyhead initially forms (figure
2,A)；Subsequent Sp5-8, successively trigger differentiation, flower glume increase for stamen, gynoecium former base inside poppyhead, finally close at one
It rises (Fig. 2, B-D).Compared with wild type, the small ear of mutant osids2 is sent out during early development compared to wild type small ear
Apparent variation is given birth to.After small ear separate living tissue is converted into floral meristem, lemma development is normal, but part glumelle is developed
It obviously latens slow, is less than wild type glumelle (Fig. 2, E-H).Part small ear glumelle former base form obviously becomes larger compared with wild type, and
Rate of development is obviously faster than wild type glumelle former base, and similar to the development of lemma former base, flower glume cannot it is closed together (Fig. 2,
I-L)。
Genetic analysis, the assignment of genes gene mapping and the map based cloning of 2 OSIDS2 gene of embodiment
Osids2 mutant is hybridized with western big 1A (being provided by Southwest University's rice research).Obtained phenotype with
The identical F of osids2 mutation type surface2Plant is used for the genetic analysis and the assignment of genes gene mapping of OSIDS2 gene.F2It is aobvious for segregating population
Show that mutant and WT lines segregation ratio meet 3：1 segregation ratio indicates osids2 mutation type surface by a pair of dominant core
Gene control.
Primary Location：It is carried out using BSA (mixing divides cluster analysis) method.Selection is uniformly distributed on 12 chromosomes of rice
480 pairs of SSR primers that Gramene, the website the Rice Genomic Research Program have been announced, in parent
Polymorphism is detected between osids2 and Xi Nong 1A.F2 is chosen for the 10 plants of wild types and 10 plant mutant body single-strain blades in segregating population
Mixed in equal amounts respectively extracts normal pond and mutation pond genomic DNA respectively, with the primer between the parent filtered out with polymorphism
Gene linkage analysis is carried out in normal and mutated gene pond.After PCR amplification and polyacrylamide gel electrophoresis and silver staining, one
As will have the single plant of maternal banding pattern be denoted as A, the single plant with male parent banding pattern is denoted as B, and the single plant with parents' banding pattern is denoted as H,
It analyzes and maps through data and convert genetic distance, screening and gene OSIDS2 chain SSR marker, discovery for recombination fraction
OSIDS2 and SSR marker S-5320 and S-3276 on the 4th chromosome long arm are chain, we have announced between S-5320 and S-3276
SSR marker carry out differential screening between parent, wherein finding three pairs of differences label S-17349, S-17407 and S-17411.With even
Lock 491 plants of F of labeled analysis2Recessive mutation single plant in segregating population, the results show that OSIDS2 gene and label S-17349, S-
Genetic distance between 17411 and S-17407 is respectively 0.1cM, 1.02cM and 2.34cM, OSIDS2 gene be located at S-17349 and
Between S-17411 (Fig. 3, A).
Finely positioning：Continue the SSR marker that the screening between S-17349 and S-17407 has parent's difference, finds two
SSR marker S-6365 and S-17391 show polymorphism between parent (S-6365, S-17391 are shown in primer sequence table 1).With this two
1464 plant new target groups are analyzed primer together with S-17349, and S-6365 and S-17391 filter out 71 and 58 respectively
Recombination individual, and S-17349 label screening goes out that 3 recombinations are individual, the recon of S-17349 and S-17391 are different, and S-
6365 recombination individual is included in S-17391, shows that OSIDS2 gene is located between SSR marker S-17349 and S-17391.
In conjunction with published Rice Genome Sequence result (http://rice.plantbiology.msu.edu/cgi-bin/
Gbrowse/rice/), physical distance is about 1.02M (Fig. 3, A) between S-17349 and S-17391.
Finely positioning section is relatively large, and annotation gene is more, and 6 transcription factor (ORF1- are had chosen after analyzing
ORF6 it) is sequenced.Separately design this six candidate gene encoder block primers (primer ORF1-F/R, ORF2-F/R, ORF3-F/
R, ORF4-F/R, GRF5-F/R, ORF6-F/R are shown in primer table 1), wild type (J10) and mutant osids2 plant are surveyed
Sequence discovery, ORF1 gene, that is, OSIDS2 gene the 198th amino acids on the 5th exon have a C base in osids2
Missing, cause frameshift mutation, the 213rd amino acids caused to be converted into terminator codon, make protein translation terminate in advance (figure
3, A；Fig. 5).According to known paddy gene group information, OSIDS2 gene belongs to myb gene family MYB-1R class transcription factor.
It is made of 6 exons, gene cDNA overall length 921bp, encoder block DNA overall length 3887bp.
In order to confirm this supposition, using real-time quantitative PCR, using Actin as internal reference, (primer Actin-F and Actin-R sequence
Column are shown in Table 1) analysis OSIDS2 gene (primer RT-OSIDS2-F and RT-OSIDS2-R sequence is shown in Table 1) in wild type and mutant
Expression in small ear.It is extracted using the RNAprep Pure Plant RNA Purification Kit kit of Tiangeng
The small ear RNA of mutant osids2 and WT lines.The total serum IgE of 1 microgram is taken to use Reagent Kit
With gDNA Eraser kit carries out reverse transcription.Quantitative PCR utilizes SYBR permix Ex Taq II Kit (TaKaRa)
Kit is expanded in Applied Biosystems 7500Real-Time PCR System, is selected in Actin conduct
Join (primer Actin-F and Actin-R sequence is shown in Table 1), each reaction carries out three repetitions and is averaged calculation expression level.Knot
Fruit shows that expression of the OSIDS2 gene in mutant children's fringe is above wild type children fringe (Fig. 3, C).
Further, it constructs OSIDS2 gene complementation carrier and carries out genetic transformation analysis.
I two restriction enzyme sites of Xba I and Kpn are chosen first on carrier pCAMBIA1300 as pCAMBIA1300 carrier
The restriction enzyme site of skeleton, and combine published Rice Genome Sequence result (http://
Rice.plantbiology.msu.edu/cgi-bin/gbrowse/rice/), and according to pEASY-Uni Seamless
Cloning and Assembly Kit recombinase specification, the recombination primer (primer of design amplification OSIDS2 genetic fragment
OSIDS2-com-F and OSIDS2-com-R is shown in 1) sequence is shown in Table.Using OSIDS2 genomic DNA as template, with primer OSIDS2-
Com-F/R amplification includes the OSIDS2 genetic fragment of promoter sequence.Again with I digestion of quick restriction enzyme Xba I and Kpn
PCAMBIA1300 carrier framework, warm bath 30-45min under the conditions of 37 DEG C are digested, electrophoresis recycles skeleton.Use pEASY-Uni
Carrier framework after digestion products that Seamless Cloning and Assembly Kit recombinase recycles glue and digestion connects
It connects and constitutes pCAMBIA1300-OSIDS2 complementing vector (Fig. 3, B), then convert osids2 mutant plants after being transferred to Agrobacterium.
In obtained transgenic plant, observe that the mutation type surface of 15 plants of transgenic plants is restored altogether, small ear
Glumelle development is normal, and internal floral organ number, form are all consistent with wild type small ear (Fig. 3, D).
The mutant phenotype of osids2 is restored in complementary plant, shows that LOC_Os04g47890 is exactly OSIDS2 gene.
The expression pattern analysis of 3 OSIDS2 gene of embodiment
Using real-time quantitative PCR (primer RT-OSIDS2-F and RT-OSIDS2-R sequence is shown in Table 1, using Actin as internal standard,
Primer Actin-F and Actin-R sequence be shown in Table 1), detection OSIDS2 gene rice growth each period (Seedling Stage,
Tiller early stage, heading stage, florescence) different parts are (root, leaf, stem, sheath, bud, auricle, section, fringe, flower glume, lodicule, stamen, female
Stamen) expression, the results showed that OSIDS2 gene has an expression in each period different parts, especially in heading early stage inflorescence
Long 0.5cm or less is expressed most strongly, and also has expression (Fig. 4, A) in each floral organ.
Further have detected the table of three flower glume characterizing genes OsMADS1, OsMADS6 and DL in different times small ear
It reaches.OsMADS1 gene is expressed in the main part (except marginal portion) of entire lemma and glumelle, determines lemma and glumelle main body
Partial feature development (Prasad et al., 2005).The expression of the gene has compared to wild type in osids2 mutant
Apparent up-regulation, this is (Fig. 4, the B) being consistent with the performance of mutant glumelle lemma, additional glume shape organ；DL gene master
It to be expressed in the middle arteries of lemma, the feature development (Yamaguchi et al., 2004) of lemma be determined, in mutant children's spike length
Obviously raised when to 0.5-2cm, this be directly translated into mutant glumelle lemma, even some lemmas lemma be consistent (Fig. 4,
C)；OsMADS6 gene, is expressed in glumelle, determines the feature development (Prasad et al., 2005) of glumelle, in mutant children
The expression of the gene is obviously lowered in fringe, this degenerates with mutant glumelle, glumelle lemma is consistent (Fig. 4, D).
The subcellular localization of 4 OSIDS2 albumen of embodiment
For the function of clear OSIDS2 albumen, OSIDS2 albumen is studied using rice protoplast transient expression system
Positioning.(primer is OEOSIDS2-GFP-F1 and OEOSIDS2-GFP-R2,1) sequence is shown in Table simultaneously for the code area of amplification OSIDS2
It is connected to 35S-GFP-NOS (pA7) expression vector (purchase of BioVector plasmid vector bacterium cell gene collection)
In, construct 35S-OEOSIDS2-GFP fusion protein carrier.Then by 35S-GFP and 35S-OEOSIDS2-GFP fusion protein
Carrier is transferred in rice protoplast respectively, and the fluorescence of GFP is observed with Olympus laser scanning confocal micro- scope.Micro-
(green fluorescence) is expressed in the entire cell in addition to vacuole to control fusion protein under the microscope, wherein red fluorescence is that leaf is green
The autofluorescence (Fig. 6, A-D) of body；And OEOSIDS2-GFP fusion protein, only observed in nucleus green fluorescence (Fig. 6,
E-H).The result shows that OSIDS2 albumen, which is one, is located in endonuclear transcription factor.
Above-described specific embodiment has carried out further the purpose of the present invention, technical scheme and beneficial effects
It is described in detail, it should be understood that being not intended to limit the present invention the foregoing is merely a specific embodiment of the invention
Protection scope, all within the spirits and principles of the present invention, any modification, equivalent substitution, improvement and etc. done should all include
Within protection scope of the present invention.
Claims (10)
1. the preparation of adjusting and controlling rice yield, it is characterised in that：Its main active is the protein of OSIDS2 gene coding, or
Express the element or carrier of OSIDS2 gene.
2. the preparation of adjusting and controlling rice yield as described in claim 1, it is characterised in that：The egg of the OSIDS2 gene coding
White matter has the amino acid sequence shown such as SEQ ID No.1 or SEQ ID No.2.
3. the preparation of adjusting and controlling rice yield as claimed in claim 2, it is characterised in that：The OSIDS2 gene has such as
SEQ ID No.3 or SEQ ID No.4 nucleotide sequence.
4. the preparation of adjusting and controlling rice grain number per spike, it is characterised in that：Its main active is the protein of OSIDS2 gene coding,
Or the element or carrier of expression OSIDS2 gene.
5. the preparation of adjusting and controlling rice grain number per spike as claimed in claim 4, it is characterised in that：The OSIDS2 gene coding
Protein has the amino acid sequence shown such as SEQ ID No.1 or SEQ ID No.2.
6. the preparation of adjusting and controlling rice grain number per spike as claimed in claim 5, it is characterised in that：The OSIDS2 gene has such as
SEQ ID No.3 or SEQ ID No.4 nucleotide sequence.
Purposes of the 7.OSIDS2 gene in adjusting and controlling rice yield.
8. purposes of the OSIDS2 gene as claimed in claim 7 in adjusting and controlling rice yield, it is characterised in that：The purposes
For adjusting and controlling rice grain number per spike.
9. purposes of the OSIDS2 gene as claimed in claim 7 or 8 in adjusting and controlling rice yield, it is characterised in that：Described
The protein of OSIDS2 gene coding has the amino acid sequence shown such as SEQ ID No.1 or SEQ ID No.2.
10. purposes of the OSIDS2 gene as claimed in claim 9 in adjusting and controlling rice yield, it is characterised in that：Described
OSIDS2 gene has such as SEQ ID No.3 or SEQ ID No.4 nucleotide sequence.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110846325A (en) * | 2019-12-09 | 2020-02-28 | 中国水稻研究所 | Rice multi-flower gene MOF1 and application of protein encoded by same |
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| CN102037125A (en) * | 2008-05-01 | 2011-04-27 | 中央研究院 | Use of rice polypeptides/nucleic acids for plant improvement |
| CN107142265A (en) * | 2017-05-17 | 2017-09-08 | 西南大学 | Paddy rice 3 spends purposes of the small ear gene LF1 in grass family cereal crops yield is improved |
| CN107603963A (en) * | 2017-10-18 | 2018-01-19 | 中国水稻研究所 | A kind of rice honeysuckle small ear gene DF1 and its coding protein and application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110846325A (en) * | 2019-12-09 | 2020-02-28 | 中国水稻研究所 | Rice multi-flower gene MOF1 and application of protein encoded by same |
| CN110846325B (en) * | 2019-12-09 | 2022-08-12 | 中国水稻研究所 | Rice multi-flower gene MOF1 and application of protein encoded by same |
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