CN106554397A - From paddy rice protein OsGRF4-M and its relevant biological material regulation and control plant organ size in application - Google Patents
From paddy rice protein OsGRF4-M and its relevant biological material regulation and control plant organ size in application Download PDFInfo
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Abstract
The invention discloses a kind of applications of protein OsGRF4-M and its relevant biological material from paddy rice in regulation and control plant organ size.The protein that the present invention is provided, available from paddy rice, is named as OsGRF4-M albumen, is following (a) or (b) or (c)：A protein that the amino acid sequence of () shown in sequence in sequence table 1 is constituted；B () is by the amino acid sequence of sequence 1 is through the replacement of one or several amino acid residues and/or disappearance and/or adds and the protein by derived from sequence 1 that seed size and/or seed length with plant and/or seed grain heavy phase are closed；C () is by the amino acid sequence of sequence 1 is through the replacement of one or several amino acid residues and/or disappearance and/or adds and by sequence 1 derived from the protein related to plant organ size.The present invention can be used to increase plant seed production, for plant breeding, particularly rice breeding, with substantial worth.
Description
Technical field
The present invention relates to the protein OsGRF4-M and its relevant biological material that derive from paddy rice are big in regulation and control plant organ
Application in little.
Background technology
Paddy rice is one of cereal crops important in the world.With reduction and the rapid growth of population of cultivated area, carry
High rice yield is of great significance to ensureing China's grain security tool.
Rice yield is determined by available tillering, grain number per spike and grain type.Rice grain shape includes the length of seed, width
Degree and thickness.Rice yield can effectively be improved by the length and width for increasing rice grain.Research plant is such as
The mechanism of what regulation and control organ size has become one of Critical policies of raising crop yield.
The content of the invention
It is an object of the invention to provide a kind of protein OsGRF4-M and its relevant biological material from paddy rice is being adjusted
Application in control plant organ size.
The protein that the present invention is provided, available from paddy rice, is named as OsGRF4-M albumen, be following (a) or (b) or
(c)：
A protein that the amino acid sequence of () shown in sequence in sequence table 1 is constituted；
B () is by the amino acid sequence of sequence 1 is through the replacement of one or several amino acid residues and/or disappearance and/or adds
Plus and the protein by derived from sequence 1 that closes of seed size and/or seed length with plant and/or seed grain heavy phase；
C () is by the amino acid sequence of sequence 1 is through the replacement of one or several amino acid residues and/or disappearance and/or adds
Plus and by sequence 1 derived from the protein related to plant organ size.
In order that the protein in (a) is easy to purifying, can be in the amino acid sequence by shown in sequence in sequence table 1
The amino terminal or the upper label as shown in table 1 of carboxyl terminal connection of the protein of row composition.
The sequence of 1 label of table
| Label | Residue | Sequence |
| Poly-Arg | 5-6 (usually 5) | RRRRR |
| Poly-His | 2-10 (usually 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag II | 8 | WSHPQFEK |
| c-myc | 10 | EQKLISEEDL |
Protein in above-mentioned (b) can be artificial synthesized, also can first synthesize its encoding gene, then carry out biological expression and obtains.
The encoding gene of the protein in above-mentioned (b) can be by will lack one in the DNA sequence dna shown in sequence in sequence table 2
Or the codon of several amino acid residues, and/or carry out the missense mutation of one or several base-pairs, and/or its 5 '
End and/or 3 ' ends connect the coded sequence of the label shown in table 1 and obtain.
The gene (OsGRF4-M genes) for encoding the OsGRF4-M albumen falls within protection scope of the present invention.
The OsGRF4-M genes concretely following DNA molecular 1) or 2) or 3) or 4) or 5)：
1) DNA molecular in sequence table shown in sequence 2；
2) seed size and/or seed length under strict conditions with the DNA sequence dna hybridization and coded plant for 1) limiting
And/or the DNA molecular of seed grain weight GAP-associated protein GAP；
3) 1) or 2) there is more than 90% homology with the DNA sequence dna for limiting and the seed size of coded plant and/or
The DNA molecular of seed length and/or seed grain weight GAP-associated protein GAP；
4) DNA under strict conditions with the DNA sequence dna hybridization and coded plant organ size GAP-associated protein GAP for 1) limiting
Molecule；
1) or 2) 5) there is more than 90% homology and the related egg of coded plant organ size to the DNA sequence dna for limiting
White DNA molecular.
Above-mentioned stringent condition can be：50 DEG C, in 7%SDS, 0.5M Na3In the mixed solution of PO4 and 1mM EDTA
Hybridization, at 65 DEG C, 0.1 × SSC is rinsed in 0.1%SDS.Above-mentioned stringent condition is alternatively：In 6 × SSC, 0.5%
In the solution of SDS, hybridize at 65 DEG C, then with 2 × SSC, 0.1%SDS and 1 × SSC, 0.1%SDS respectively wash film one
It is secondary.
Recombinant expression carrier containing the OsGRF4-M genes, expression cassette, transgenic cell line or recombinant bacterium are belonged to
Protection scope of the present invention.OsGRF4-M genes insertion expression vector can be obtained by the recombinant expression carrier
Recombinant plasmid.The expression vector concretely carrier pMDC43.The preparation method of the recombinant expression carrier is concrete such as
Under：(1) the OsGRF4-M genes are connected with carrier pCR8/GW/TOPO, obtain recombinant plasmid TOPO-OsGRF4-M；
(2) recombinant plasmid TOPO-OsGRF4-M and carrier pMDC43 carries out LR recombining reactions, obtains containing the OsGRF4-M
The recombinant plasmid of gene, is named as recombinant plasmid pMDC43-OsGRF4-M.
The primer pair for expanding the total length or its any fragment of the OsGRF4-M genes falls within protection scope of the present invention.
The present invention also protects a kind of method for cultivating genetically modified plants, is by the OsGRF4-M channel genes purpose plant
In, obtain the genetically modified plants of seed and/or seed grain weight and/or seed length more than the purpose plant.It is described
OsGRF4-M genes specifically can import the purpose plant by the recombinant expression carrier.It is in methods described, described heavy
Group expression vector can by using Ti-plasmids, Ri plasmids, plant viral vector, directly delivered DNA, microinjection,
The conventional biology methods such as conductance, agriculture bacillus mediated convert plant cell or tissue, and the plant tissue of conversion is cultivated
Into plant.The OsGRF4-M genes specifically can import the purpose plant by recombinant plasmid pMDC43-OsGRF4-M.
The purpose plant is monocotyledon or dicotyledon.The monocotyledon can be grass, specifically may be used
For paddy rice, such as in rice varieties, spend No. 11.
The present invention also protects the OsGRF4-M albumen, the OsGRF4-M genes, the recombinant expression carrier, table
Up to box, transgenic cell line or recombinant bacterium or methods described, seed is big and/or seed length is big and/or seed cultivating
Application in the great plant of grain.The plant is monocotyledon or dicotyledon.The monocotyledon can be
Grass, concretely paddy rice, such as spend No. 11 in rice varieties.
The present invention also protects the OsGRF4-M albumen in the seed size and/or seed length and/or kind of regulation and control plant
Application in seed weight.The plant is monocotyledon or dicotyledon.The monocotyledon can be grass family
Plant, concretely paddy rice, such as spend No. 11 in rice varieties.
Described in any of the above, seed is alternatively referred to as seed.
Grain described in any of the above heavy concretely mass of 1000 kernel or 100-grain weight.
The present inventor has cloned OsGRF4-M genes from huge Large grain rice, finds its coded sequence with middle flower
11 have compared some places' differences, will spend in 11, can spend 11 in making in OsGRF4-M channel genes to wild-type variety
Seed significantly increase.The present invention can be used to increase plant seed production, for plant breeding, particularly rice breeding,
With substantial worth.
Description of the drawings
Structural representations of the Fig. 1 for carrier pCR8/GW/TOPO.
Structural representations of the Fig. 2 for carrier pMDC43.
Fig. 3 is the partial results of PCR identifications.
Fig. 4 is the result of OsGRF4-M gene expression amounts analysis.
Fig. 5 is seed photo.
Results of the Fig. 6 for seed length.
Specific embodiment
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method, if no special instructions, is conventional method.Test material used in following embodiments, if no special instructions,
It is from routine biochemistry reagent shop what is be commercially available.Quantitative test in following examples, is respectively provided with three repetitions real
Test, results averaged.
Carrier pCR8/GW/TOPO：Invitrogen.The structural representation of carrier pCR8/GW/TOPO is shown in Fig. 1.
Refer to the bibliography of " carrier pMDC43 "：Curtis MD,Grossniklaus U(2003)A gateway
cloning vector set for high-throughput functional analysis of genes in planta.
Plant Physiol.133:462-469)；The public can be obtained from Chinese Academy of Sciences's heredity with Developmental Biology research,
The biomaterial only attach most importance to duplicate invention related experiment used by, can not use as other purposes.Carrier pMDC43
Structural representation see Fig. 2.
Refer to the bibliography of " Agrobacterium tumefaciems GV3101 "：Li,Y.,Zheng,L.,Corke,F.,Smith,C.,
and Bevan,M.W.(2008)Control of final seed and organ size by the DA1gene family
in Arabidopsis thaliana.Genes Dev 22,1331-1336；The public can be hereditary from the Chinese Academy of Sciences and be sent out
Educate biological study to be obtained, the biomaterial only attach most importance to duplicate invention related experiment used by, can not as other use
Way uses.
Rice varieties " in spend No. 11 " (spend 11 in referred to as)：Zhu Xudong, Chen Hongqi, Luo Da, Zhang Jianjun, Fang Hongmin, Min
Continue pattern；The separation and identification of 11 radiomutants are spent in paddy rice；Rice in China science, 2003,17 (3)：205-210..
Inducing culture：Organic+2,4-D the 2.5mg/L of N6 a great number of elements+MS-Fe salt+B5 trace element+B5
+ proline 500mg/L+glutamine 500mg/L+CH 300mg/L+ sucrose 30g/L+Gelrite
2.6mg/L, pH 5.8.
Subculture medium：Organic+2,4-D the 2.0mg/L of N6 a great number of elements+MS-Fe salt+B5 trace element+B5
+ proline 500mg/L+glutamine 500mg/L+CH 300mg/L+ sucrose 30g/L+Gelrite
2.6mg/L, pH 5.8.
Co-culture culture medium (solid)：Organic+the 2,4-D of N6 a great number of elements+MS-Fe salt+B5 trace element+B5
2.0mg/L+CH 500mg/L+ inositols 2000mg/L+AS 100 μM+sucrose 30g/L+Gelrite 2.6mg/L,
pH 5.5。
Co-culture culture medium (liquid)：Organic+the 2,4-D of N6 a great number of elements+MS-Fe salt+B5 trace element+B5
2.0mg/L+CH 500mg/L+ inositols 2000mg/L+AS 100 μM+sucrose 30g/L, pH 5.5.
Screening and culturing medium：Organic+2,4-D the 2.0mg/L of N6 a great number of elements+MS-Fe salt+B5 trace element+B5
+ proline 500mg/L+glutamine 500mg/L+CH 300mg/L+ sucrose 30g/L+Gelrite
2.6mg/L+cef 250mg/L+Hyg 50mg/L, pH 5.8.
Differential medium：Organic+NAA the 0.1mg/L+ of N6 a great number of elements+MS-Fe salt+B5 trace element+B5
KT 4mg/L+proline 500mg/L+glutamine 500mg/L+CH 300mg/L+ sucrose 30g/L+
Gelrite 2.6mg/L+cef 250mg/L+Hyg 50mg/L, pH 5.8.
Root media：1/2N6 a great number of elements+MS-Fe salt+B5 trace elements+sucrose 30g/L+Agar
0.8% (weight/mass percentage composition), pH 5.8.
The discovery of embodiment 1, OsGRF4-M albumen and its encoding gene
1st, the total serum IgE reverse transcription for extracting huge Large grain rice is cDNA.
2nd, as template, the primer pair constituted using OsGRF4-S and OsGRF4-A is carried out the cDNA obtained with step 1
PCR is expanded.
OsGRF4-S:5'-ATGACGATGCCGTATGCCTC-3’；
OsGRF4-A:5'-TCAGTCACCATTAGTTGATCG-3’。
3rd, the pcr amplification product that step 2 is obtained is entered into row agarose gel electrophoresis and reclaims the band of about 1185bp.
ORFs in pcr amplification product shown in the sequence 2 with sequence table, the ORFs coded sequence
Protein shown in the sequence 1 of table.
Protein shown in the sequence 1 of sequence table is named as into OsGRF4-M albumen, by coding OsGRF4-M albumen
Unnamed gene is OsGRF4-M genes.In spend 11 in OsGRF4-M albumen homology highest protein such as sequence tables
Sequence 3 shown in (be named as OsGRF4 albumen, existing sequence), the sequence 4 of its coded sequence such as sequence table
Shown (being named as OsGRF4 genes, existing sequence).
The acquisition and identification of embodiment 2, genetically modified plants
First, the preparation of recombinant plasmid
1st, the double chain DNA molecule shown in the sequence 2 of composition sequence table.
2nd, the double chain DNA molecule obtained with step 1 is as template, the primer constituted using OsGRF4-S and OsGRF4-A
To entering performing PCR amplification, pcr amplification product is obtained.
OsGRF4-S:5'-ATGACGATGCCGTATGCCTC-3’；
OsGRF4-A:5'-TCAGTCACCATTAGTTGATCG-3’。
3rd, the pcr amplification product that step 2 is obtained is entered into row agarose gel electrophoresis and reclaims the fragment of about 1185bp.
4th, the fragment for reclaiming step 3 is connected with carrier pCR8/GW/TOPO, obtains recombinant plasmid TOPO-OsGRF4-M.
According to sequencing result, structure is carried out to recombinant plasmid TOPO-OsGRF4-M and is described as follows：In carrier pCR8/GW/TOPO
AttL1 and attL2 between insert OsGRF4-M genes shown in the sequence 2 of sequence table.
5th, recombinant plasmid TOPO-OsGRF4-M and carrier pMDC43 carry out LR recombining reactions, obtain containing ordered list
The recombinant plasmid of the OsGRF4-M genes shown in sequence 2, is named as recombinant plasmid pMDC43-OsGRF4-M.Root
According to sequencing result, structure is carried out to recombinant plasmid pMDC43-OsGRF4-M and is described as follows：By the attR1 of carrier pMDC43
Small fragment and attR1 between replaces in order to little between the attL1 and attL2 of recombinant plasmid TOPO-OsGRF4-M
Fragment.
The restructuring matter of the OsGRF4 genes shown in the sequence 4 containing ordered list is prepared with reference to the method for step 1 to 5
Grain, is named as recombinant plasmid pMDC43-OsGRF4.
2nd, turn the acquisition of OsGRF4-M gene plants
1st, recombinant plasmid pMDC43-OsGRF4-M is imported into Agrobacterium tumefaciems GV3101, obtains recombinational agrobacterium.
2nd, 11 are spent in the recombinational agrobacterium conversion for obtaining step 1, concrete operations are as follows：
(1) Fiber differentiation of Mature Embryos of Rice callus
1. 11 mature seeds are spent in removing shell, are carried out surface sterilizing and (first 1-2min, Ran Houyong are soaked with 70% ethanol
0.1% mercuric chloride soaks 10min), then seed is placed on suck dry moisture on aseptic filter paper by then aseptic water washing 3-4 time.
2. after completing step 1., seed is placed on inducing culture flat board, 26 DEG C of light cultures 10-15 days.
3. after completing step 2., the callus that mature embryo scultellum grows is peeled, being placed on subculture medium flat board is carried out
(squamous subculture is once every two weeks for squamous subculture；26 DEG C of light cultures), select squamous subculture 5-7 days, pale yellow more
Injured tissue carries out step (3).
(2) prepare bacteria suspension
The recombinational agrobacterium that step 1 is obtained is suspended in liquid to co-culture in culture medium, adjustment bacteria concentration is
OD600nm=0.3-0.5, is subsequently adding acetosyringone and makes its concentration be 100mM.
(3) callus that step (1) is obtained is placed in the bacteria suspension that step (2) is obtained, room temperature places 20min
And rock frequently, then take out callus and be placed on aseptic filter paper to suck unnecessary bacterium solution, then callus is turned
Move on to and be covered with the co-cultivation culture medium flat plate of one layer of aseptic filter paper, 26 DEG C of light cultures 2-3 days.
(4), after completing step (3), callus is placed on screening and culturing medium flat board and 26 DEG C of light cultures 14 days,
Then callus is transferred on new screening and culturing medium flat board 26 DEG C of light cultures 14 days.Most of callus exists
10 days or so brownings after screening, then regrow out milky resistant calli at the edge of browning tissue.
(5), after completing step (4), the fine and close resistant calli of milk yellow is selected, differential medium flat board is placed in
On, first light culture 3 days is then transferred to cultivate under 15h/d illumination conditions.Culture has green pointing out in 15-25 days or so
Existing, culture further differentiated seedling after 30-40 days.
(6) in step (5), when seedling bud is about 2cm, transfers them to,
High about 10cm, the seedling of well developed root system are selected, culture medium is washed away, is transplanted to field, as T0 generations intend turning OsGRF4-M
Gene plant.
Obtain 35 plants of T0 generation plans and turn OsGRF4-M gene plants.
3rd, turn the identification of OsGRF4-M gene plants
(1) 35 plants of T0 generation plans that step 2 is obtained are turned into OsGRF4-M gene plants and enters performing PCR identification.
Extract the genomic DNA of the fresh blade of plant and as template, the primer pair constituted with K4S-F and K4S-R
Enter performing PCR amplification, PCR primer HindIII carries out digestion, and (OsGRF4-M genes can be cut, OsGRF4 bases
Because being cut).
K4S-F：5’-CCCTTTCAAGCTATTCTAAGC-3’；
K4S-R：5’-TTCCCAAAGAACGAAAGTGG-3’.
Partial detection is shown in Fig. 3.Turn OsGRF4-M gene plants because while containing OsGRF4-M genes and OsGRF4
Gene, has two bands Jing after digestion.In spend 11 to comprise only OsGRF4 genes, it is impossible to it is digested, therefore only one
The single band of bar.In Fig. 1, swimming lane 1-17 is part T0 generations to intend turning OsGRF4-M gene plants, during swimming lane 18 is
Spend 11.
In 35 plants of T0 generations, intend turning in OsGRF4-M gene plants, and 28 plants is to turn OsGRF4-M gene plants.
(2) T0 generations are turned OsGRF4-M gene plants carries out OsGRF4-M gene expression amount analyses.
Extract the total serum IgE of the fresh blade of plant and reverse transcription is cDNA, then by real-time PCR
The expression of OsGRF4-M genes in (Lightcycler 480, ROCHE) detection plant.Fluorescent dye is
Lightcycler 480SYBR Green I Master(ROCHE).Internal reference is ACTIN1.For detecting OsGRF4-M
The primer pair of gene is made up of K4RT-F and K4RT-R.For detecting the primer of ACTIN1 by ACTIN1F and ACTIN1R
Composition.
K4RT-F：5’-ATTCCAAGTACTGCGAGCGC-3’；
K4RT-R：5’-GGCGACCAGCTGCGTTT-3’.
ACTIN1F：5’-TGCTATGTACGTCGCCATCCAG-3’；
ACTIN1R：5’-AATGAGTAACCACGCTCCGTCA-3’.
The relative expression quantity of OsGRF4-M genes is shown in Fig. 4 (1 spends 11,2 in turn OsGRF4-M gene plants).
3rd, turn the acquisition of OsGRF4 gene plants
Replace recombinant plasmid pMDC43-OsGRF4-M to carry out step 2 with recombinant plasmid pMDC43-OsGRF4, turned
OsGRF4 gene plants.
4th, phenotypic evaluation
In T0 generations, turn OsGRF4-M gene plants and grow under the natural conditions of field, collect seed, observe and clap after maturation
According to (28 plants of T0 generations turn OsGRF4-M gene plants, the length of 100 seeds of every plant of random measurement to measurement seed length
Degree, results averaged), (in 28 plants of T0 generations, turn OsGRF4-M gene plants, every plant of random measurement to statistics mass of 1000 kernel
The grain of 100 seeds weighs and is scaled mass of 1000 kernel, results averaged).
In T0 generations, turn OsGRF4 gene plants and grow under the natural conditions of field, collect seed, observe and take pictures after maturation,
(in 10 plants of T0 generations, turn OsGRF4 gene plants, the length of 100 seeds of every plant of random measurement, knot to measurement seed length
Fruit is averaged), (in 10 plants of T0 generations, turn OsGRF4 gene plants, 100 kinds of every plant of random measurement to statistics mass of 1000 kernel
The grain of son weighs and is scaled mass of 1000 kernel, results averaged).
In spend 11 under the natural conditions of field grow, maturation after collect seed, observe and take pictures, measure seed length (10
11 are spent in strain, the length of 100 seeds of every plant of random measurement, results averaged), statistics mass of 1000 kernel is (in 10 plants
The grain of 11,100 seeds of every plant of random measurement is spent to weigh and be scaled mass of 1000 kernel, results averaged).
Above-mentioned each plant carries out parallel test, i.e., cultivate under conditions of identical.Using body formula mirror (LEICA
S8APO, Germany) observe and take pictures (LEICA DFC420, Germany).It is long using Image J1.41 softwares measurement seed
Degree.Statistical analysis is carried out using EXCEL.
Photo is shown in Fig. 5, two, the left side be it is middle spend 11 seed, two, the right is to turn OsGRF4-M gene plants in T0 generations
Seed, scale is 1mm.
The result of seed length is shown in Fig. 6,1 for it is middle spend 11 seed, 2 is to turn OsGRF4-M gene plants in T0 generations
Seed.
The result of seed length and mass of 1000 kernel the results are shown in Table 2 (mean+SDs).
The phenotype statistics of 2 genetically modified plants of table
| It is middle spend 11 seed | In T0 generations, turn the seed of OsGRF4-M gene plants | In T0 generations, turn the seed of OsGRF4 gene plants | |
| Seed length (mm) | 6.72±0.03 | 9.94±0.09** | 7.81±0.04** |
| Mass of 1000 kernel (g) | 25.61±0.17 | 34.56±0.08** | 28.92±0.32** |
Note：It is notable that * represents 0.01 level differences of P ＜.
Spend in compared with the of 11, the seed length and mass of 1000 kernel for turning OsGRF4-M gene plants is dramatically increased.With turn
OsGRF4 gene plants are compared, and the seed length and mass of 1000 kernel for turning OsGRF4-M gene plants is dramatically increased.
The above results show that OsGRF4-M genes are proceeded to by the organ size positive correlation of OsGRF4-M genes and plant
After expressing in purpose plant, the organ size of plant can be increased.
Claims (10)
1. a kind of protein, is following (a) or (b) or (c)：
A protein that the amino acid sequence of () shown in sequence in sequence table 1 is constituted；
B () is by the amino acid sequence of sequence 1 is through the replacement of one or several amino acid residues and/or disappearance and/or adds
Plus and the protein by derived from sequence 1 that closes of seed size and/or seed length with plant and/or seed grain heavy phase；
C () is by the amino acid sequence of sequence 1 is through the replacement of one or several amino acid residues and/or disappearance and/or adds
Plus and by sequence 1 derived from the protein related to plant organ size.
2. the gene of protein described in claim 1 is encoded.
3. gene as claimed in claim 2, it is characterised in that：The gene for it is following 1) or 2) or 3) or 4)
Or DNA molecular 5)：
1) DNA molecular in sequence table shown in sequence 2；
2) seed size and/or seed length under strict conditions with the DNA sequence dna hybridization and coded plant for 1) limiting
And/or the DNA molecular of seed grain weight GAP-associated protein GAP；
3) 1) or 2) there is more than 90% homology with the DNA sequence dna for limiting and the seed size of coded plant and/or
The DNA molecular of seed length and/or seed grain weight GAP-associated protein GAP；
4) DNA under strict conditions with the DNA sequence dna hybridization and coded plant organ size GAP-associated protein GAP for 1) limiting
Molecule；
1) or 2) 5) there is more than 90% homology and the related egg of coded plant organ size to the DNA sequence dna for limiting
White DNA molecular.
4. the recombinant expression carrier, expression cassette, transgenic cell line or restructuring containing gene described in Claims 2 or 3
Bacterium.
5. the total length or the primer pair of its any fragment of gene described in Claims 2 or 3 are expanded.
6. a kind of method for cultivating genetically modified plants, be by channel genes purpose plant described in Claims 2 or 3,
Obtain the genetically modified plants of seed and/or seed grain weight and/or seed length more than the purpose plant.
7. method as claimed in claim 6, it is characterised in that：The purpose plant is monocotyledon or dicotyledonous
Plant.
8. method as claimed in claim 7, it is characterised in that：The monocotyledon is paddy rice.
9. protein described in claim 1, or, gene described in Claims 2 or 3, or, described in claim 4
Recombinant expression carrier, expression cassette, transgenic cell line or recombinant bacterium, or, the method described in claim 7 or 8,
Application in the plant that seed is big and/or seed length is big and/or seed grain is great is cultivated.
10. protein described in claim 1 is regulating and controlling the seed size and/or seed length and/or seed grain weight of plant
In application.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2015106406634 | 2015-09-30 | ||
| CN201510640663 | 2015-09-30 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106554397A true CN106554397A (en) | 2017-04-05 |
| CN106554397B CN106554397B (en) | 2019-08-30 |
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| WO2019158911A1 (en) * | 2018-02-14 | 2019-08-22 | Institute Of Genetics And Developmental Biology Chinese Academy Of Sciences | Methods of increasing nutrient use efficiency |
| CN110157730A (en) * | 2019-05-29 | 2019-08-23 | 福建省农业科学院生物技术研究所 | A method of it releasing miRNA and function is inhibited to promote expression of target gene |
| CN111087457A (en) * | 2019-12-31 | 2020-05-01 | 中国科学院遗传与发育生物学研究所 | Protein NGR5 for improving nitrogen utilization rate and crop yield, and coding gene and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107937416A (en) * | 2017-12-29 | 2018-04-20 | 中国科学院遗传与发育生物学研究所 | Improve gene and its application of nitrogen fertilizer for paddy rice utilization ratio and yield |
| CN107937416B (en) * | 2017-12-29 | 2020-11-03 | 中国科学院遗传与发育生物学研究所 | Gene for improving utilization efficiency and yield of rice nitrogen fertilizer and application thereof |
| WO2019158911A1 (en) * | 2018-02-14 | 2019-08-22 | Institute Of Genetics And Developmental Biology Chinese Academy Of Sciences | Methods of increasing nutrient use efficiency |
| CN111818794A (en) * | 2018-02-14 | 2020-10-23 | 中国科学院遗传与发育生物学研究所 | Method for increasing nutrient utilization efficiency |
| US11873499B2 (en) | 2018-02-14 | 2024-01-16 | Institute Of Genetics And Developmental Biology Chinese Academy Of Sciences | Methods of increasing nutrient use efficiency |
| CN110157730A (en) * | 2019-05-29 | 2019-08-23 | 福建省农业科学院生物技术研究所 | A method of it releasing miRNA and function is inhibited to promote expression of target gene |
| CN110157730B (en) * | 2019-05-29 | 2021-06-08 | 福建省农业科学院生物技术研究所 | Method for relieving miRNA inhibition function and promoting target gene expression |
| CN111087457A (en) * | 2019-12-31 | 2020-05-01 | 中国科学院遗传与发育生物学研究所 | Protein NGR5 for improving nitrogen utilization rate and crop yield, and coding gene and application thereof |
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