CN108948170B - Plant type growth and development related protein and coding gene and application thereof - Google Patents

Abstract

The invention discloses a plant type growth and development related protein, and a coding gene and application thereof. The protein provided by the invention is named as OsSGD2 protein and is a protein shown in a sequence 1 of a sequence table. The gene coding the OsSGD2 protein is named as OsSGD2 gene and also belongs to the protection scope of the invention. The invention also provides a method for preparing a transgenic plant, which comprises the following steps: a transgenic plant having a reduced plant height compared to the starting plant is obtained by introducing a substance inhibiting the expression of the OsSGD2 gene into the starting plant. The invention not only provides a basis for further clarifying the molecular mechanism of the rice plant type, but also provides new gene resources and breeding resources for rice breeding. The invention has important application value for effectively regulating and controlling the rice plant type by utilizing the gene resource through genetic breeding and genetic engineering methods.

Description

Plant type growth and development related protein and coding gene and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a plant type growth and development related protein, and a coding gene and application thereof.

Background

Rice is one of the important grain crops in China, and the rice production plays an extremely important role in the grain production and the national economy in China. Due to rapid development of economy, increased population pressure and reduced arable land area, rice production is under tremendous pressure. Therefore, the cultivation of new rice varieties with significantly improved yield potential will contribute an important force for alleviating or solving the problem of food safety. The plant type is an important agronomic character closely related to the rice yield, and mainly depends on several aspects such as plant height, leaf type, tillering number, tillering angle, spike morphology and the like. In recent years, research on rice plant type functional genes by using methods of molecular genetics and functional genomics has become a hotspot and has important practical significance.

The individual comprehensive agronomic character of the gene for controlling the rice plant height character discovered and identified at present is poor in performance and difficult to use in production, sd1 is applied to production for nearly 60 years, and wide utilization of a single gene can influence the diversity differentiation of rice. The main problem of plant type breeding at present is that the number of applicable plant height genes is too small, and it is very important to break through the bottleneck in applying new plant height genes. Therefore, new resources for plant height are explored and identified, a molecular mechanism for regulating the plant height is deeply explored, dependence on a single plant height gene is avoided in a breeding process of a rice breeder, meanwhile, a theoretical basis can be provided for molecular design breeding and rice plant leaf type improvement, and efficient breeding is finally realized.

Disclosure of Invention

The invention aims to provide a plant type growth and development related protein, and a coding gene and application thereof.

The protein provided by the invention is derived from rice (Oryza sativa), is named as OsSGD2 protein, and is (a) or (b) or (c) or (d) as follows:

(a) protein shown in a sequence 1 in a sequence table;

(b) protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence 1, is related to plant types and is derived from the plant types;

(c) a protein derived from rice and having 95% or more identity and the same function as (a);

(d) a fusion protein obtained by attaching a tag to the N-terminus or/and the C-terminus of (a), (b) or (C).

The labels are specifically shown in table 1.

TABLE 1 sequences of tags

The protein can be synthesized artificially, or can be obtained by synthesizing the coding gene and then carrying out biological expression.

The gene coding the OsSGD2 protein is named as OsSGD2 gene and also belongs to the protection scope of the invention.

The OsSGD2 gene is the DNA molecule as shown in the following (1) or (2) or (3) or (4) or (5):

(1) the coding region is a DNA molecule shown as a sequence 2 in a sequence table;

(2) DNA molecule shown in sequence 3 in the sequence table;

(3) a DNA molecule which has more than 75% of identity with (1) and codes plant type related protein;

(4) a DNA molecule which is derived from rice, has more than 90% of identity with (1) or (2) and encodes a plant type related protein;

(5) and (3) a DNA molecule which is hybridized with the protein (1) or (2) under strict conditions and encodes a plant type related protein of the plant.

The stringent conditions are hybridization and washing of the membrane 2 times 5min at 68 ℃ in a solution of 2 XSSC, 0.1% SDS and 2 times 15min at 68 ℃ in a solution of 0.5 XSSC, 0.1% SDS.

The above-mentioned identity of 75% or more may be 80%, 85%, 90% or 95% or more.

The expression cassette, the recombinant vector or the recombinant bacterium containing the OsSGD2 gene belong to the protection scope of the invention.

The recombinant expression vector containing the gene can be constructed by using the existing expression vector. When the gene is used for constructing a recombinant expression vector, any one of enhanced, constitutive, tissue-specific or inducible promoters can be added in front of the transcription initiation nucleotide, and can be used alone or combined with other plant promoters; in addition, when the gene is used to construct a recombinant expression vector, enhancers, including translational or transcriptional enhancers, may be used, and these enhancer regions may be ATG initiation codons or adjacent regions initiation codons, etc., but must be in the same reading frame as the coding sequence to ensure proper translation of the entire sequence. The translational control signals and initiation codons are widely derived, either naturally or synthetically. The translation initiation region may be derived from a transcription initiation region or a structural gene. In order to facilitate identification and screening of the transgenic plant or the transgenic microorganism, an expression vector to be used may be processed, for example, a gene for expressing an enzyme or a luminescent compound which produces a color change in the plant or the microorganism, a gene for an antibiotic marker having resistance or a chemical-resistant agent marker, etc. From the viewpoint of safety of transgenes, the transformed plants or microorganisms can be directly screened phenotypically without adding any selectable marker gene.

The plant type is particularly the plant height.

The invention also provides a method for preparing a transgenic plant, which comprises the following steps: a transgenic plant having a reduced plant height compared to the starting plant is obtained by introducing a substance inhibiting the expression of the OsSGD2 gene into the starting plant. The substance inhibiting the expression of the OsSGD2 gene can be specifically an interference vector. The starting plant is a dicotyledonous plant or a monocotyledonous plant. The monocot may be rice, such as rice Kitaake.

The invention also provides a plant breeding method, which comprises the following steps: the activity and/or content of the OsSGD2 protein in the target plant is reduced, so that the plant height of the plant is reduced. The target plant is a dicotyledonous plant or a monocotyledonous plant. The monocot may be rice, such as rice Kitaake.

The invention also protects the application of the substance for inhibiting the expression of the OsSGD2 gene, and is used for cultivating transgenic plants with reduced plant height. The substance inhibiting the expression of the OsSGD2 gene can be specifically an interference vector. The plant is a dicotyledonous plant or a monocotyledonous plant. The monocot may be rice, such as rice Kitaake.

The invention also protects the application of the substance for reducing the activity and/or content of the OsSGD2 protein, and aims to reduce the plant height of plants. The plant is a dicotyledonous plant or a monocotyledonous plant. The monocot may be rice, such as rice Kitaake.

The invention also protects the application of the OsSGD2 protein, which is (I), (II) or (III):

regulating and controlling the plant type of the plant;

(II) regulating and controlling the plant height of the plant;

(III) increasing the plant height of the plant.

The plant is a dicotyledonous plant or a monocotyledonous plant. The monocot may be rice, such as rice Kitaake.

The invention also protects an interference fragment which is a DNA molecule and has a fragment A and a fragment B, wherein the fragment A and the fragment B are reversely complementary, and the fragment A is shown as nucleotides from 1 st to 531 th sites of a sequence 2 in a sequence table. The interference fragment is specifically shown as a sequence 4 in a sequence table.

The invention also protects an interference vector.

Any one of the interference vectors has a fragment A and a fragment B, wherein the fragment A and the fragment B are reversely complementary, and the fragment A is shown as nucleotides 1 to 531 in a sequence 2 of a sequence table. Any one of the interference vectors has a DNA molecule shown in a sequence 4 of a sequence table.

In the above, the transgenic rice is understood to include not only the first generation transgenic rice obtained by transforming the recipient rice with the gene, but also the progeny thereof. For transgenic rice, the gene can be propagated in the species, or transferred into other varieties of the same species, including commercial varieties in particular, using conventional breeding techniques. The transgenic rice comprises seeds, callus, complete plants and cells.

The invention firstly identifies the protein OsSGD2 related to the growth and development of the rice plant type, and the coding gene of the protein OsSGD2 related to the growth and development of the rice plant type can cause the dwarfing of the rice plant height under the condition of function loss or expression quantity reduction, thereby proving that the protein related to the growth and development of the rice plant type or the gene thereof plays an important role in controlling the height of the rice plant. The invention not only provides a basis for further clarifying the molecular mechanism of the rice plant type, but also provides new gene resources and breeding resources for rice breeding. The transgenic rice with the OsSGD2 gene expression reduced, which is obtained by the invention, is used as a new rice germplasm material, and can be used for researching the molecular mechanism of rice dwarfing and finding more genes for regulating and controlling the plant height development of rice. The invention has important application value for effectively regulating and controlling the rice plant type by utilizing the gene resource through genetic breeding and genetic engineering methods.

Drawings

FIG. 1 shows the expression level of OsSGD2 gene.

FIG. 2 is a photograph of a plant at the heading stage.

FIG. 3 shows the plant height of plants in the mature period.

Detailed Description

The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.

The vector pLHRNAi used in the examples is a pLHRNAi recombinant vector constructed in example 1 of the specification of a patent with the application number of 201110055864.X (publication number of CN 102191262B, publication date of 2013-03-20).

N6 medium: sigma, catalog No. C1416.

Rice Kitaake (also called wild-type rice, denoted by WT), is described in the following documents: gao H, Zheng XM, Fei G, Chen J, Jin M, Ren Y, Wu W, Zhou K, Sheng P, Zhou F, Jiang L, Wang J, Zhang X, Guo X, Wang JL, Cheng Z, Wu C, Wang H, Wan JM. ehd4 codes a novel and Oryza-gene-specific regulator of photoperiodic flowing in rice.OS GENET.2013, 9(2): e 3281. MT plants are naturally occurring mutant of OsSGD2 gene mutation obtained from a very large number of rice Kitaake plants, and theoretically have the same phenotype as RNAi plants as a control of RNAi plants.

A new protein is found from rice Kitaake and is named as OsSGD2 protein, and the new protein is shown as a sequence 1 in a sequence table. The gene coding the OsSGD2 protein is named as OsSGD2 gene, an open reading library in cDNA is shown as a sequence 2 in a sequence table, and the gene in genome DNA is shown as a sequence 3 in the sequence table.

Example 1 obtaining and identification of interfering plants

Construction of interference vectors

1. Total RNA of rice Kitaake seedlings in 14 days is extracted and reverse transcription is carried out to obtain cDNA.

2. And (3) carrying out PCR amplification by using the cDNA obtained in the step 1 as a template and a primer consisting of OsSGD2-sense-F and OsSGD2-sense-R to obtain a fragment 1(SEQ forward direction).

OsSGD2-sense-F：5'-TTCTGCACTAGGTACCAGGCCTGATGATGGGGGCCACTTCTCCGT-3'；

OsSGD2-sense-R：5'-CTGACGTAGGGGCGATAGAGCTCGTGCGGCGAGAGCACCGTGGGC-3'。

3. And (3) carrying out PCR amplification by using the cDNA obtained in the step (1) as a template and a primer consisting of OsSGD2-antisense-F and OsSGD2-antisense-R to obtain a fragment 2(SEQ reverse direction).

OsSGD2-antisense-F:5'-CGGGGATCCGTCGACTACGTGCGGCGAGAGCACCGTGGGC-3'；

OsSGD2-antisense-R:5'-AGGTGGAAGACGCGTTACATGATGGGGGCCACTTCTCCGT-3'。

4. The vector pLHRNAi is cut by restriction endonuclease SacI to obtain a linearized vector.

5. And (3) integrating the fragment 1 obtained in the step (2) and the linearized vector obtained in the step (4) by adopting a homologous recombination directional cloning method to obtain a recombinant vector.

6. And (3) carrying out enzyme digestion on the recombinant vector obtained in the step (5) by using restriction enzyme SnaBI to obtain a linearized vector.

7. And (3) integrating the fragment 2 obtained in the step (3) and the linearized vector obtained in the step (6) by adopting a homologous recombination directional cloning method to obtain a recombinant plasmid pLHRNAi-OsSGD 2.

Sequencing is carried out on the recombinant plasmid pLHRNAi-OsSGD2, and sequencing results show that the recombinant plasmid has a DNA molecule shown as a sequence 4 in a sequence table. In the sequence 4 of the sequence table, a fragment A and a fragment B are provided, the fragment A and the fragment B are reversely complementary, and the fragment A is shown as nucleotides 1 to 531 of the sequence 2 of the sequence table.

Second, obtaining of RNAi interfering plant

1. The recombinant plasmid pLHRNAi-OsSGD2 is introduced into Agrobacterium tumefaciens EHA105 to obtain recombinant Agrobacterium tumefaciens.

2. Suspending the recombinant Agrobacterium obtained in step 1 with a liquid N6 medium containing 100. mu.M acetosyringone to obtain OD_600nmAnd (4) a bacterium solution which is approximately equal to 0.5, namely the staining solution.

3. Immersing mature embryogenic callus of rice Kitaake in the infection solution obtained in the step 2 for 30min, then taking the callus, sucking dry the bacterial solution by using filter paper, then placing the callus on a solid N6 culture medium, and culturing for 3 days at 24 ℃.

4. After completion of step 3, the callus was placed on solid N6 medium containing 150mg/L hygromycin and cultured at 24 ℃ for 16 days.

5. After completion of step 4, the calli were cultured on solid N6 medium containing 200mg/L hygromycin, subcultured every 15 days.

6. And (3) placing the callus obtained in the step (5) on a differentiation culture medium containing 150mg/L hygromycin, culturing for 45 days at 24 ℃ (the height of the overground part of the plant is about 15cm), opening a bottle mouth, hardening seedlings for 3 days, and transplanting the plant to a greenhouse for cultivation, namely a T0 generation plant.

Differentiation medium: taking 2mg of 6-BA, 0.2mg of NAA, N64 g, 1g of hydrolyzed casein, 0.1g of inositol, 25g of sucrose, 2.4g of sorbitol and 7g of agar powder, dissolving with deionized water and fixing the volume to 1L.

7. And (4) carrying out PCR identification on the T0 generation plants obtained in the step 6.

Genomic DNA of plant seedlings is extracted, forward PCR identification is carried out by adopting a primer pair consisting of 1390-F and FAD2-R, and reverse PCR identification is carried out by adopting a primer pair consisting of 1390-R and FAD 2-F.

1390-F：5’-TGCCTTCATACGCTATTTATTTGC-3’；

FAD2-R：5’-GAAGCGACGGACCTGGAGAT-3’。

FAD2-F：5’-CCTTTCACAACCTGATTTCCCA-3’；

1390-R：5’-TAATCATCGCAAGACCGGCAACAGG-3’。

If the forward PCR identification and the reverse PCR identification both show the amplification product, the plant is an RNAi interference plant.

The obtained RNAi interfering plants are named as RNAi-1 plants, RNAi-2 plants, … … and RNAi-n plants in sequence. n is a natural number.

And thirdly, identifying the expression level of the OsSGD2 gene.

The plants to be tested are respectively as follows: RNAi-1 plant, RNAi-2 plant, rice Kitaake plant, MT plant.

Extracting total RNA of the plant to be detected and carrying out reverse transcription to obtain cDNA. And (3) detecting the relative expression quantity of the OsSGD2 gene by using cDNA as a template, using Ubiquitin gene as an internal reference gene and adopting fluorescent quantitative PCR.

The primers used to identify the Ubiquitin gene were as follows:

UBI-F：5’-GCTCCGTGGCGGTATCAT-3’；

UBI-R：5’-CGGCAGTTGACAGCCCTAG-3’。

primers for identifying the OsSGD2 gene were as follows:

OSSGD2-qRT-F：5’-GGAGGCCTTGGCGATCAA-3’；

OSSGD2-qRT-R:5’-ACTCCATCTCCGTCTGCTTC-3’。

the results are shown in FIG. 1. Compared with rice Kitaake plants, the expression level of OsSGD2 gene in RNAi-1 plants and RNAi-2 plants is obviously reduced.

Fourth, obtaining empty carrier plants

Replacing the recombinant plasmid pLHRNAi-OsSGD2 with the vector pLHRNAi, and performing the other steps to obtain a transgenic empty vector plant.

Fifth, phenotypic identification

The plants to be tested are respectively as follows: RNAi-1 plant, RNAi-2 plant, empty vector plant, MT plant, rice Kitaake plant.

The plant to be tested is planted in the field, cultured under parallel conditions, and the phenotype of the plant in the whole growth period is observed.

Photographs of the plants at the heading node stage are shown in FIG. 2. Compared with the Kitaake plants, the RNAi-1 plants and the RNAi-2 plants have the phenotype of plant height dwarfing. The phenotype of the empty vector transferred plant is consistent with that of the rice Kitaake plant.

The plant height of the plants in the mature period is counted, and the result is shown in figure 3. Compared with the Kitaake plants, the plant heights of the RNAi-1 plants and the RNAi-2 plants in the mature period are remarkably reduced by over 60 percent. Compared with the Kitaake plants, the plant height of the empty carrier plants has no obvious difference.

The results show that the OsSGD2 protein participates in controlling the growth and development of rice, and is specifically related to plant type, more specifically related to plant height.

SEQUENCE LISTING

<110> institute of crop science of Chinese academy of agricultural sciences

<120> plant type growth and development related protein, and coding gene and application thereof

<130>GNCYX181611

<160>4

<170>PatentIn version 3.5

<210>1

<211>229

<212>PRT

<213>Oryza sativa

<400>1

Met Met Gly Ala Thr Ser Pro Ser Gly Leu Glu Leu Thr Met Ala Val

1 5 10 15

Pro Gly Leu Ser Ser Ser Gly Ser Glu Gly Ala Gly Cys Asn Asn Asn

20 25 30

Asn Ala Gly Gly Gly Cys Asn Met Arg Asp Leu Asp Ile Asn Gln Pro

35 40 45

Ala Ser Gly Gly Glu Glu Glu Glu Phe Pro Met Gly Ser Val Glu Glu

50 55 60

Asp Glu Glu Glu Arg Gly Val Gly Gly Pro His Arg Pro Lys Lys Leu

65 70 75 80

Arg Leu Ser Lys Glu Gln Ser Arg Leu Leu Glu Glu Ser Phe Arg Leu

85 90 95

Asn His Thr Leu Thr Pro Lys Gln Lys Glu Ala Leu Ala Ile Lys Leu

100 105 110

Lys Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln Asn Arg Arg Ala

115 120 125

Arg Thr Lys Leu Lys Gln Thr Glu Met Glu Cys Glu Tyr Leu Lys Arg

130 135 140

Cys Phe Gly Ser Leu Thr Glu Glu Asn Arg Arg Leu Gln Arg Glu Val

145 150 155 160

Glu Glu Leu Arg Ala Met Arg Val Ala Pro Pro Thr Val Leu Ser Pro

165 170 175

His Thr Arg Gln Pro Leu Pro Ala Ser Ala Leu Thr Met Cys Pro Arg

180 185 190

Cys Glu Arg Ile Thr Ala Ala Thr Gly Pro Pro Ala Val Arg Pro Pro

195 200 205

Pro Ser Ser Ala Ala Ala Ala Ala Pro Ser Pro Phe His Pro Arg Arg

210 215 220

Pro Ser Ala Ala Phe

225

<210>2

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<213>Oryza sativa

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agggacctgg acatcaacca gccggcgagc ggcggcgagg aggaggagtt cccgatgggc 180

agcgtggagg aggacgagga ggagaggggc gtcggtgggc cccaccgccc caagaagctc 240

cgcctctcca aggagcagtc ccgcctcctc gaggagagct tccgcctcaa ccataccctc 300

acgccgaagc aaaaggaggc cttggcgatc aaactgaagc tgcggccgag gcaggtggag 360

gtctggtttc agaaccgtag ggcaaggacg aagctgaagc agacggagat ggagtgcgag 420

tacctgaagc gctgcttcgg gtcgctgacg gaggagaacc gccggctgca gcgggaggtg 480

gaggagctgc gggcgatgcg ggtggccccg cccacggtgc tctcgccgca caccaggcag 540

ccgctcccgg cgtccgcgct caccatgtgc ccccgctgcg agcgcatcac cgccgccacc 600

ggcccgcctg ccgtgcgccc gccgccgtcg tcagccgccg ccgccgcccc ctcgcccttc 660

caccctcgcc gcccctctgc ggccttctag 690

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ggcctcagct cctctggttc aggtaagctc gggagacaac caatctccta gctatgtctc 180

catcctacaa acgtgcgtgc cttgatcttt tttttttcgt cggaacatgc ccatatgtga 240

gagcgcacct ccaacctttt gaagttggag ttaccaacac atatatgcat ggtcttcaga 300

atgtttttct cggcatggag actgaacttg tttgcggttt tcattgacgt ttgcttctgg 360

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ttgtttgcag aaggggccgg ttgcaacaac aacaacgccg gtggcggctg caacatgagg 480

gacctggaca tcaaccagcc ggcgagcggc ggcgaggagg aggagttccc gatgggcagc 540

gtggaggagg acgaggagga gaggggcgtc ggtgggcccc accgccccaa gaagctccgc 600

ctctccaagg agcagtcccg cctcctcgag gagagcttcc gcctcaacca taccctcacg 660

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tggtacatgt acctgcatca gaatctagtt cttgatagaa ttagtgcttc gtgctcctcg 840

atcataatca cagatcgagc tcactagttc gtactaactg gcgtgtggat ttgggtcttg 900

aagtggagca ctcctgctga cggactgttg gacattgaaa gggcgtggta tatgaattga 960

ttccgagtaa atggattcgg aggaccatcg atcattggat ggtactcctc cataattgct 1020

cggtgcttga tgatatggtc cttgcacggg agatatacta gtacttggtg tctgtgtgca 1080

cagccgcaca gctataaaag gagaggaggc acctctgcac cctctctctc tctctctctc 1140

tcgcacacgt cacaccactt cagggttatg ctgcataaaa acgtctactg agtactagca 1200

gcactatgca gtagttctat aatgtttgtt ctgtgcagtg gtttactggt atttacgtgc 1260

tgtacagctg atagtataat gaactatata tagtgcttta gatacttcat aacaacgtct 1320

actgaagtat atcctagcag cactatgcag tataaagttg gctttatgca gctatactgg 1380

tatatttacg tacagtaaag cacagtagag agtaatgcag gagtatatga taaactagtg 1440

cctggtaatg cagttaaggc tcgtttgact agcaggggtg ttactgcgat ttactactat 1500

actacgaccc ttgcaatatg gagtatactg agatactctg aacctgaagg cctgcatgtg 1560

taatggcaag caatttaatg cgttggaaaa tgtaacctct gcagaagcaa aaggaggcct 1620

tggcgatcaa actgaagctg cggccgaggc aggtggaggt ctggtttcag aaccgtaggg 1680

caaggtaaaa aatcccccat cttgatgcaa tatatagagt acccaataat aagttcacaa 1740

gcttctttat ttatcatttc agtagaatct actcctcagt cctcgttaca ctatcatata 1800

ctagagtatt ttctccgcta taaatagcta tgttctgtac ataccaaggc aaagacttgt 1860

acacgagaat gtgacgaaga agcatacttg atcctatata tgttagccat acatagaagt 1920

attagcattt gaaatggttc atgtgacaaa acttcacaaa agacaaaagg cattagagta 1980

cagtcaatcc ggcttatgct aaatgatgat cttgaaagcg accactatat ggcatttctt 2040

tacaaatttg cattagagcc tacgtaaaag ctagattgat tactacactt attagtgttg 2100

gtgcaacaca agttatacta gcaggaccac aatgaaccca tcattccaac aaaattatgc 2160

aatcttttcc ctcacgaagg tgacaaggat ccttaggtat ccatcttgaa actagtagtg 2220

catccaagat gtgagacaac aaacaattag ctagattctc tagcctgata aagtgtaggc 2280

agttccatca tcacttatga attgctgcta gatggcgtag attactccat gatactcgtt 2340

cggtttgatt ccaggattaa aaaaaaccga gttgcttgtt ccaatggcgc cggcacgtgg 2400

actaatcggc acgtgctaca gctctctgta actttgaccc tagctaccta atgcgaaatc 2460

ttagtttaac acgtcccgtg aaggatggcg tacaacttgt aacctgaacg ccaatccatt 2520

tcaattgatc cctacaaagg ccattgcctt cactctacgt cgccgtcgag gtgacaatga 2580

tatcggaatg gacgcctaag gtcaaattga gtaactactc tagttcatgc cgctatttta 2640

atctgcaatc aagtagtact gagctactaa taatagtaat gtcgcgcagt taacttcggc 2700

aagttcatgg cagaaacatc ttttcatttg ttaacgctca atcgctgccc gattgttact 2760

gatcgtaacg tacgtggcag aacagaaagt gatgggagct ggtggctgtg tgcaggacga 2820

agctgaagca gacggagatg gagtgcgagt acctgaagcg ctgcttcggg tcgctgacgg 2880

aggagaaccg ccggctgcag cgggaggtgg aggagctgcg ggcgatgcgg gtggccccgc 2940

ccacggtgct ctcgccgcac accaggcagc cgctcccggc gtccgcgctc accatgtgcc 3000

cccgctgcga gcgcatcacc gccgccaccg gcccgcctgc cgtgcgcccg ccgccgtcgt 3060

cagccgccgc cgccgccccc tcgcccttcc accctcgccg cccctctgcg gccttctagg 3120

cccaaggtct cgcatcctaa aggccacaaa aagatgggcc tcgccgtccc gtgtaaattg 3180

aaggagacga cgcagcggta gagggccccg tgttggcgtt gttgcgttgg gtccagtcgt 3240

ctcagaggag tcagaggcgt ggacaaaatg tttgttgcat tgtttgtgtt ccattggtcg 3300

ttgatcttag cttgatgatc tgtgattcgc gccttgctcg tctaggtttc tcttgggccg 3360

acgcatgatc ttttttcccc gtgtcggacc gtattgggcc ttttgagatc gacgggcccc 3420

ctccttgttt cgcatgtgga catgttattt cgtccttttg ggccaacaat tcagcacggt 3480

ggccaacggt gcgtagttcg tcgtcgtttt ctttc 3515

<210>4

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<212>DNA

<213>Artificial sequence

<400>4

ttgatatact tggatgatgg catatgcagc agctatatgt ggattttttt agccctgcct 60

tcatacgcta tttatttgct tggtactgtt tcttttgtcg atgctcaccc tgttgtttgg 120

tgttacttct gcactaggta ccaggcctga tgatgggggc cacttctccg tcaggcctgg 180

agctcaccat ggctgtcccc ggcctcagct cctctggttc agaaggggcc ggttgcaaca 240

acaacaacgc cggtggcggc tgcaacatga gggacctgga catcaaccag ccggcgagcg 300

gcggcgagga ggaggagttc ccgatgggca gcgtggagga ggacgaggag gagaggggcg 360

tcggtgggcc ccaccgcccc aagaagctcc gcctctccaa ggagcagtcc cgcctcctcg 420

aggagagctt ccgcctcaac cataccctca cgccgaagca aaaggaggcc ttggcgatca 480

aactgaagct gcggccgagg caggtggagg tctggtttca gaaccgtagg gcaaggacga 540

agctgaagca gacggagatg gagtgcgagt acctgaagcg ctgcttcggg tcgctgacgg 600

aggagaaccg ccggctgcag cgggaggtgg aggagctgcg ggcgatgcgg gtggccccgc 660

ccacggtgct ctcgccgcac cagctctatc gcccctacgt cagctccatc tccaggtccg 720

tcgcttctct tccatttctt ctcattttcg attttgattc ttatttcttt ccagtagctc 780

ctgctctgtg aatttctccg ctcacgatag atctgcttat actccttaca ttcaacctta 840

gatctggtct cgattctctg tttctctgtt tttttctttt ggtcgagaat ctgatgtttg 900

tttatgttct gtcaccatta ataataatga actctctcat tcatacaatg attagtttct 960

ctcgtctaca aaacgatatg ttgcattttc acttttcttc tttttttcta agatgatttg 1020

ctttgaccaa tttgtttaga tctttatttt attttatttt ctggtgggtt ggtggaaatt 1080

gaaaaaaaaa aaaacagcat aaattgttat ttgttaatgt attcattttt tggctatttg 1140

ttctgggtaa aaatctgctt ctactattga atctttcctg gattttttac tcctattggg 1200

tttttatagt aaaaatacat aataaaagga aaacaaaagt tttatagatt ctcttaaacc 1260

ccttacgata aaagttggaa tcaaaataat tcaggatcag atgctctttg attgattcag 1320

atgcgattac agttgcatgg caaattttct agatccgtcg tcacatttta ttttctgttt 1380

aaatatctaa atctgatata tgatgtcgac aaattctggt ggcttataca tcacttcaac 1440

tgttttcttt tggctttgtt tgtcaacttg gttttcaata cgatttgtga tttcgatcgc 1500

tgaattttta atacaagcaa actgatgtta accacaagca agagatgtga cctgccttat 1560

taacatcgta ttacttacta ctagtcgtat tctcaacgca atcgtttttg tatttctcac 1620

attatgccgc ttctctactc tttattcctt ttggtccacg cattttctat ttgtggcaat 1680

ccctttcaca acctgatttc ccactttgga tcatttgtct gaagactctc ttgaatcgtt 1740

accacttgtt tcttgtgcat gctctgtttt ttagaattaa tgataaaact attccatagt 1800

cttgagtttt cagcttgttg attcttttgc ttttggtttt ctgcagaaac atgggtgcag 1860

gtggaagacg cgttacgtgc ggcgagagca ccgtgggcgg ggccacccgc atcgcccgca 1920

gctcctccac ctcccgctgc agccggcggt tctcctccgt cagcgacccg aagcagcgct 1980

tcaggtactc gcactccatc tccgtctgct tcagcttcgt ccttgcccta cggttctgaa 2040

accagacctc cacctgcctc ggccgcagct tcagtttgat cgccaaggcc tccttttgct 2100

tcggcgtgag ggtatggttg aggcggaagc tctcctcgag gaggcgggac tgctccttgg 2160

agaggcggag cttcttgggg cggtggggcc caccgacgcc cctctcctcc tcgtcctcct 2220

ccacgctgcc catcgggaac tcctcctcct cgccgccgct cgccggctgg ttgatgtcca 2280

ggtccctcat gttgcagccg ccaccggcgt tgttgttgtt gcaaccggcc ccttctgaac 2340

cagaggagct gaggccgggg acagccatgg tgagctccag gcctgacgga gaagtggccc 2400

ccatcatgga tccccgggaa ttctaagagg agtccaccat ggtagatctg actagtgtta 2460

acgctagcca ccaccaccac caccacgtgt gaattacagg tgaccagctc gaatttcccc 2520

gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg 2580

atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgta 2636

Claims (5)

1. A method of making a transgenic plant comprising the steps of: introducing a substance inhibiting the expression of a gene encoding an OsSGD2 protein into a starting plant to obtain a transgenic plant with a reduced plant height compared with the starting plant;

the OsSGD2 protein is (a) or (d) as follows:

(a) protein shown in a sequence 1 in a sequence table;

(d) a fusion protein obtained by connecting a label to the N terminal or/and the C terminal of (a);

the plant is rice.

2. The method of claim 1, wherein:

the gene coding the OsSGD2 protein is a DNA molecule as described in the following (1) or (2):

(1) the coding region is a DNA molecule shown as a sequence 2 in a sequence table;

(2) DNA molecule shown in sequence 3 in the sequence table.

3. The application of a substance for inhibiting the expression of a gene coding the OsSGD2 protein in cultivating transgenic plants with reduced plant height;

the OsSGD2 protein is (a) or (d) as follows:

(a) protein shown in a sequence 1 in a sequence table;

(d) a fusion protein obtained by connecting a label to the N terminal or/and the C terminal of (a);

the plant is rice.

4. Use according to claim 3, characterized in that:

the gene coding the OsSGD2 protein is a DNA molecule as described in the following (1) or (2):

(1) the coding region is a DNA molecule shown as a sequence 2 in a sequence table;

(2) DNA molecule shown in sequence 3 in the sequence table.

5, application of the OsSGD2 protein in regulating and controlling plant height of plants;

the OsSGD2 protein is (a) or (d) as follows:

(a) protein shown in a sequence 1 in a sequence table;

(d) a fusion protein obtained by connecting a label to the N terminal or/and the C terminal of (a);

the plant is rice.

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