CN108948166B

CN108948166B - Plant height related protein IbCBEFP and coding gene and application thereof

Info

Publication number: CN108948166B
Application number: CN201810884103.7A
Authority: CN
Inventors: 刘庆昌; 翟红; 何绍贞; 赵宁; 任志彤
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2018-08-06
Filing date: 2018-08-06
Publication date: 2021-01-05
Anticipated expiration: 2038-08-06
Also published as: CN108948166A

Abstract

The invention discloses a plant height related protein IbCBEFP, and a coding gene and application thereof. The protein provided by the invention is obtained from sweet potatoes (Ipomoea batatas), is named as IbCBEFP protein and is the protein shown in a sequence 1 in a sequence table. Nucleic acid molecules encoding such proteins are also within the scope of the invention. The invention also protects the application of the IbCBEFP protein: regulating and controlling the plant height of the plant; promoting plant dwarfing; the plant height is promoted to be reduced; promoting the plant root length to be shortened; reducing the auxin content; reducing the gibberellin content of plants. The invention also protects the application of the nucleic acid molecule: cultivating transgenic plants with changed plant height; cultivating a dwarfing transgenic plant; cultivating transgenic plants with reduced plant height; cultivating transgenic plants with shortened root length; cultivating transgenic plants having reduced auxin content; transgenic plants with reduced gibberellin content are grown. The invention has wide application space and market prospect in the agricultural field.

Description

Plant height related protein IbCBEFP and coding gene and application thereof

Technical Field

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

Background

The growth and development of plants is an extremely complex process, which is based on the metabolism of various substances, and is manifested as the processes of germination, rooting, leaf growth, further maturation of plant bodies, flowering, fruiting, and finally senescence and death. The growth is mainly cell division and elongation, and the development is a process of simple and complex change of the structure and function of the plant body caused by differentiation of cells, tissues and organs in the life history of the whole plant. The series of processes are the result of space-time specific expression of related genes under the action of internal and external factors.

The plant type is the morphological characteristics and spatial arrangement mode of plants, and is the comprehensive agronomic characters, which influence the planting density of crops, and further influence the yield and economic efficiency. Reasonable plant type can greatly improve yield. The plant height is a key factor in plant type, higher plants are easy to fall down and influence the high yield of the sweet potatoes, so that the short vines are the requirements of the high yield and stress resistance of the sweet potatoes and the requirements of mechanized production. In actual production, the dwarfing plant can enable the stem and the vine of the plant to be upright, is more favorable for photosynthesis, is more favorable for mechanical operation in the harvesting process, saves the cost and improves the operation efficiency.

Disclosure of Invention

The invention aims to provide a plant height related protein IbCBEFP, and a coding gene and application thereof.

The protein provided by the invention is obtained from sweet potatoes (Ipomoea batatas), is named as IbCBEFP protein, and is (a1) or (a2) or (a3) or (a 4):

(a1) protein shown as a sequence 1 in a sequence table;

(a2) a fusion protein obtained by attaching a tag to the N-terminus or/and the C-terminus of the protein represented by (a 1);

(a3) protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in (a1) and is related to plant types;

(a4) a protein derived from sweetpotato and having 98% or more identity to (a 1).

The labels may be as shown in table 1.

TABLE 1

Label (R)	Residue of	Sequence of
			Poly-Arg	5-6 (usually 5)	RRRRR
Poly-His	2-10 (generally 6)	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tagⅡ	8	WSHPQFEK
			c-myc	10	EQKLISEEDL

The protein can be artificially synthesized, or can be obtained by synthesizing the coding gene and performing biological expression. The coding gene of the protein can be obtained by deleting one or more codons of amino acid residues in a DNA sequence shown in a sequence 2 in a sequence table, and/or carrying out missense mutation of one or more base pairs, and/or connecting a coding sequence of a label shown in the table 1 at the 5 'end and/or the 3' end.

Nucleic acid molecules encoding such proteins are also within the scope of the invention.

The nucleic acid molecule is (b1) or (b2) or (b3) as follows:

(b1) the coding region is a DNA molecule shown as a sequence 2 in a sequence table;

(b2) a DNA molecule derived from sweetpotato and having 95% or more identity to (b1) and encoding the protein;

(b3) a DNA molecule which hybridizes under stringent conditions with the nucleotide sequence defined in (b1) and encodes said protein.

The stringent conditions can be hybridization and washing with 0.1 XSSPE (or 0.1 XSSC), 0.1% SDS solution at 65 ℃ in DNA or RNA hybridization experiments.

Expression cassettes, recombinant vectors or recombinant microorganisms containing the nucleic acid molecules are within the scope of the invention.

The invention also protects the application of the IbCBEFP protein, which is (c1), (c2), (c3), (c4), (c5) or (c 6):

(c1) regulating and controlling the plant height of the plant;

(c2) promoting plant dwarfing;

(c3) the plant height is promoted to be reduced;

(c4) promoting the plant root length to be shortened;

(c5) reducing the auxin content;

(c6) reducing the gibberellin content of plants.

The invention also protects the application of the nucleic acid molecule, which is (d1), (d2), (d3), (d4), (d5) or (d 6):

(d1) cultivating transgenic plants with changed plant height;

(d2) cultivating a dwarfing transgenic plant;

(d3) cultivating transgenic plants with reduced plant height;

(d4) cultivating transgenic plants with shortened root length;

(d5) cultivating transgenic plants having reduced auxin content;

(d6) transgenic plants with reduced gibberellin content are grown.

The invention also provides a method for preparing a transgenic plant, which comprises the following steps: introducing the nucleic acid molecule into a starting plant to obtain a transgenic plant with reduced plant height and/or shortened root length.

The invention also provides a plant breeding method, which comprises the following steps: increasing the content and/or activity of the IbCBEFP protein in the target plant, thereby reducing the plant height and/or root length of the plant.

The invention also provides a method for preparing a transgenic plant, which comprises the following steps: introducing the nucleic acid molecule into a starting plant to obtain a transgenic plant with reduced auxin and/or gibberellin content.

The invention also provides a plant breeding method, which comprises the following steps: increasing the content and/or activity of the IbCBEFP protein in the target plant, thereby reducing the content of auxin and/or gibberellin in the plant.

Any of the above plants is a monocot or a dicot. The dicotyledonous plant can be sweet potato, such as chestnut of sweet potato variety.

The IbCBEFP gene is over-expressed in the sweet potato, so that the shape of the plant can be changed, the plant height and the root length are obviously reduced, and the contents of auxin and gibberellin are also obviously reduced. Therefore, the IbCBEFP protein and the coding gene thereof have important theoretical significance and practical value in the process of regulating and controlling the growth and development of plants. The invention has wide application space and market prospect in the agricultural field.

Drawings

FIG. 1 is an electrophoretogram of a part of a plant.

Fig. 2 is the relative expression level of IbCbEFP gene.

FIG. 3 shows the results of plant height.

FIG. 4 shows the results of root length.

FIG. 5 shows the auxin content in fresh root/stem/leaf samples.

FIG. 6 shows the gibberellin content in the fresh root/stem/leaf samples.

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.

Sweet potato variety Lushu No. 3, abbreviated as Lushu No. 3, is described in the following documents: constructing a Dianthus haichii, Shangli, Liuqingchang, sweet potato stem nematode induced inhibition hybrid cDNA library and analyzing an expression sequence label, report of agricultural biotechnology, 2010,18 (1): 141-148.

sweet potato variety chestnut flavor, abbreviated chestnut flavor, is described in the following documents: yu B, ZHai H, Wang YP, Zang N, He SZ, Liu QC.efficient Agrobacterium tumefaciens-mediated transformation using organized genetic subspecies in sweet pototo, Ipomoea batatas (L.) Lam.2007, Plant Cell, Tissue & Organ Culture, 90(3): 265-273.

The vector pCAMBIA3301 is a product of Cambia corporation. The vector pBI121 is a product of Clontech.

Example 1 obtaining of plant height-related protein IbCBEFP and encoding Gene thereof

1. Taking the No. 3 aseptic seedling of the Lushu potato and spreading leaf blades.

2. And (3) taking the leaves obtained in the step (1), and extracting total RNA by using a plant total RNA extraction kit.

3. And (3) taking the total RNA obtained in the step (2), and carrying out reverse transcription by using a QuantScript RT Kit Quant cDNA Kit to obtain cDNA.

4. And 3, designing and artificially synthesizing primers GSP-1 and GSP-2 by taking the cDNA obtained in the step 3 as a template, amplifying by using a RACE method to obtain a 3' -RACE fragment of about 400bp, and sequencing.

GSP-1：5′-GTACTCCAAATTACAATGCTGGA-3′；

GSP-2：5′-GTGGGCCATTTAAGACATCAT-3′。

5. And 3, designing and artificially synthesizing primers GSP-3 and GSP-4 by taking the cDNA obtained in the step 3 as a template, amplifying by using a RACE method to obtain a 5' -RACE fragment of about 200bp, and sequencing.

GSP-3：5′-5′-TAGGTTGACCAGAAGTAGCCATTG-3′；

GSP-4：CCAGAAGATACAGGGTTTGGAGAT-3′。

6. Splicing the nucleotide sequence of the 3 '-RACE fragment and the nucleotide sequence of the 5' -RACE fragment into candidate genes, further designing and artificially synthesizing primers O-F and O-R according to the candidate genes, carrying out PCR amplification by taking the cDNA obtained in the step 3 as a template, obtaining an amplification product and sequencing.

O-F：5′-ATGGCTGCTCCGAATATGG-3′；

O-R：5′-TTAAAACGCTTTCCAGCTATC-3′。

The sequencing result shows that the PCR amplification product is shown as a sequence 2 in the sequence table, and the protein shown as a sequence 1 in the coding sequence table.

The protein shown in the sequence 1 of the sequence table is named as IbCBEFP protein. The gene encoding IbCbEFP protein was designated IbCbEFP gene.

Example 2 application of IbCBEFP protein to influence plant type

Construction of recombinant plasmid pCB-IbCBEFP

1. Synthesizing a double-stranded DNA molecule shown in a sequence 2 in a sequence table.

2. And (3) carrying out PCR amplification by using the DNA molecules obtained in the step (1) as a template and adopting a primer pair consisting of OE-F-BglII and OE-R-PmlI.

OE-F-BglII：5′-GAAGATCTATGGCTGCTCCGAATATGG-3′；

OE-R-PmlI：5′-GCCACGTGTTAAAACGCTTTCCAGCTATC-3′。

3. And (3) recovering the PCR amplification product obtained in the step (2), performing double enzyme digestion by using restriction enzymes BglII and PmlI, and recovering the enzyme digestion product.

4. The vector pCAMBIA3301 was double-digested with restriction enzymes HindIII and EcoRI, and the vector backbone of about 11256bp was recovered.

5. The vector pBI121 was digested with restriction enzymes HindIII and EcoRI to recover a fragment of about 3412bp (i.e., a GUS gene expression cassette having, in order, a 35S promoter, a GUS gene and a NOS terminator).

6. And (4) connecting the fragment obtained in the step (5) with the vector skeleton obtained in the step (4) to obtain the recombinant plasmid pCBGUS.

7. The recombinant plasmid pCBGUS is double digested with restriction enzymes BglII and PmlI, and the vector skeleton of about 12660bp is recovered.

8. And (4) connecting the enzyme digestion product obtained in the step (3) with the vector framework obtained in the step (7) to obtain a recombinant plasmid pCB-IbCBEFP. According to the sequencing result, the structure of the recombinant plasmid pCB-IbCBEFP is described as follows: the vector pCAMBIA3301 is used as a starting vector, a GUS gene expression cassette is inserted between HindIII and EcoRI enzyme cutting sites, and an IbCBEFP gene is inserted between BglII and PmlI enzyme cutting sites.

Second, obtaining transgenic plant of sweet potato

1. And (3) introducing the recombinant plasmid pCB-IbCBEFP into the Agrobacterium tumefaciens EHA105 to obtain the recombinant Agrobacterium tumefaciens.

2. Resuspending the recombinant Agrobacterium obtained in step 1 with a liquid MS medium containing 2.0mg/L2, 4-D to obtain OD_600nmAbout 0.8 bacterial suspension.

3. Adding the chestnut flavor embryo suspended cell mass with the diameter of 0.7-1.4mm into the bacterial suspension obtained in the step 2, immersing for 5min, transferring the cell mass onto a co-culture medium, and culturing in the dark at 26-28 ℃ for 3 d.

Co-culture medium: solid MS culture medium containing 30mg/L AS (acetosyringone).

4. After completion of step 3, the cell pellet was collected, washed with the delayed culture medium, and then cultured in the delayed culture medium at 26-28 ℃ for 1 week in the dark.

Delayed culture medium: liquid MS culture medium containing 2 mg/L2, 4-D and 300mg/L cephamycin.

5. After completion of step 4, the cell pellet was collected, transferred to selection medium, and cultured in the dark at 26-28 ℃ for 8 weeks (transferred to new selection medium every 2 weeks).

Screening a culture medium: solid MS culture medium containing 2 mg/L2, 4-D, 300mg/L cefamycin and 0.25mg/L glufosinate-ammonium.

6. After step 5, the callus with good growth state is collected and transferred to an induction medium to be cultured until turning green. The culture conditions are as follows: at 26-28 deg.C, 13 hr light (illumination intensity 3000Lx)/11 hr dark. It usually takes 2-4 weeks.

Induction medium: solid MS culture medium containing 1.0mg/L ABA and 300mg/L cephamycin.

7. After step 6 is completed, the green mature somatic embryos are transferred to a solid MS culture medium and cultured for 4-8 weeks to obtain complete regeneration plants. The culture conditions are as follows: at 26-28 deg.C, 13 hr light (illumination intensity 3000Lx)/11 hr dark.

8. And (3) performing GUS staining identification on the regenerated plants: leaves were taken and stained with GUS, and if blue, GUS positive was indicated.

9. Performing PCR identification on GUS positive regeneration plants: extracting genome DNA of plant leaves, carrying out PCR amplification by adopting a primer pair consisting of 35S-F and IbCBEFP-T-R, and then carrying out 1% agarose gel electrophoresis, wherein if a specific band of about 3741bp is displayed, the PCR identification is positive.

35S-F：5′-GACCTAACAGAACTCGCCGTA-3′；

IbCbEFP-T-R：5′-GGAGTTGGTAGAGCCATACATTG-3′。

The electrophoretogram of the plant parts is shown in FIG. 1. In FIG. 1, M is DNA molecule Marker, L1 to L11 represent different regenerated plants, WT represents a chestnut plant, P represents a positive control (recombinant plasmid pCB-IbCBEFP), and W represents a negative control (water).

L1-L11 are all plants with IbCBEFP transgene.

10. Detecting the relative expression level of the IbCBEFP gene in the IbCBEFP transgenic plant: extracting total RNA of plant leaves and carrying out reverse transcription to obtain cDNA, taking the cDNA as a template, taking sweet potato beta-actin gene as an internal reference gene, and identifying the relative expression level of the IbCBEFP gene by adopting qRT-PCR.

Primers used to detect the IbCbEFP gene were as follows:

IbCbEFP-q-F：5′-CTGGGTCCATCTTTTGACTCCGTT-3′；

IbCbEFP-q-F：5′-TGTGTGAATCCAGCATTGTAACTTG-3′。

the primers used for detecting the β -actin gene were as follows:

Ibactin-F：5′-AGCAGCATGAAGATTAAGGTTGTAGCAC-3′；

Ibactin-R：5′-TGGAAAATTAGAAGCACTTCCTGTGAAC-3′。

the relative expression levels of IbCbEFP gene are shown in figure 2. In FIG. 2, L1 to L11 represent different regenerated plants, respectively, and WT represents a chestnut-flavored plant. In each IbCBEFP transgenic plant, the relative expression level of the IbCBEFP gene is obviously higher than that of a chestnut fragrant plant.

11. Expanding propagation of IbCBEFP transgenic plants: the L3 plant and the L5 plant were subjected to stem expansion (a common asexual propagation method for sweet potato), respectively, to obtain expanded plants.

Thirdly, characteristic identification of IbCBEFP transgenic sweet potato plants

The stem segments of the L3 plant (denoted by L3), L5 plant (denoted by L5) and chestnut plant (denoted by WT) were each operated as follows: culturing the stem segment in solid MS culture medium for 6 weeks (when the stem segment grows into plant), transplanting the plant into nutrition pot (containing culture medium comprising nutrient soil and vermiculite in equal volume), culturing for 4 weeks (13 h light/11 h dark and 3000lx light intensity every day), measuring plant height and root length, collecting root/stem/leaf, and detecting auxin (IAA) and Gibberellin (GA)₃) And (4) content. Three biological replicates were set up.

The results of plant height are shown in FIG. 3. The results of root length are shown in FIG. 4. The results show that compared with chestnut-flavor plants, the plant type of the transgenic plants is obviously dwarfed, and the plant height and the root length are obviously reduced.

Hormones are closely related to the growth and development of plants. Auxin is mainly to promote the growth of plants, mainly the growth of cells, especially the elongation of cells. Auxin acts in multiple sites, is mainly involved in the formation of cell walls and the metabolism of nucleic acids, and also promotes the biosynthesis of proteins. Generally, the polar transport from the synthesis site to the action is performed, and the promoting action on the stem, bud and root varies depending on the concentration. The content of auxin (IAA) is detected by adopting an indoleacetic acid content test kit (IAA-3-T) of Ming Biotech, Inc. of Jiangsu Ke. Detection of Gibberellins (GA)₃) The content is determined by adopting gibberellin content testing kit (GA-3-T) of Jiangsu Keming Biotech Co. Gibberellin is a highly effective plant growth regulator, and most typically plays an important role in significantly promoting the elongation growth of plant stem nodes and in physiological activities of plants from seed germination to flowering. The auxin content in the fresh root/stem/leaf samples is shown in FIG. 5. The gibberellin content of the fresh root/stem/leaf samples is shown in FIG. 6. The results show that compared with chestnut plantsIAA content and GA of transgenic plants₃The content is remarkably reduced.

SEQUENCE LISTING

<110> university of agriculture in China

<120> plant height related protein IbCBEFP, and coding gene and application thereof

<130> GNCYX181679

<160> 2

<170> PatentIn version 3.5

<210> 1

<211> 1246

<212> PRT

<213> Ipomoea batatas

<400> 1

Met Ala Ala Pro Asn Met Asp Gln Phe Glu Ala Tyr Phe Arg Arg Ala

1 5 10 15

Asp Leu Asp Gln Asp Gly Arg Ile Ser Gly Ala Glu Ala Val Ser Phe

20 25 30

Leu Gln Gly Ser Asn Leu Pro Lys Gln Val Leu Ala Gln Ile Trp Met

35 40 45

Tyr Ala Asp Gln Ser Gln Thr Gly Phe Leu Ser Arg Gln Asp Phe Tyr

50 55 60

Asn Ala Leu Lys Leu Val Thr Val Ala Gln Ser Lys Arg Glu Leu Thr

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Pro Asp Ile Val Lys Ala Ala Leu Phe Gly Pro Ala Ser Ala Lys Ile

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Pro Ala Pro Gln Ile Asn Leu Ala Ala Ile Pro Gly Pro Gln Pro Asn

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Asn Met Ser Asn Ser Pro Val Pro Ser Val Gly Ala Gly Val Pro Ala

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Ala Gly Pro Ser Ser Gly Thr Arg Gly His Gln Val Phe Gln Pro Gln

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Gln Ser Gln Leu Ala Arg Pro Pro Arg Pro Pro Ala Pro Ser Thr Thr

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Phe Gln Ser His Pro Ala Val Ser Gly Pro Gly Val Pro Val Gly Ser

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Thr Met Thr Ser Ser Asn Ser Pro Ile Ser Pro Asp Met Asn Gly Gly

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Arg Thr Ser Gly Ser Gln Pro Gly Val Thr Pro Gln Leu Ser Tyr Arg

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Gly Ile Ser Pro Lys Ser His Asp Gly Phe Gly Leu Val Ala Ser Gly

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Ser Thr Pro Pro Gln Ser Lys Pro Gln Asp Ala Ala Leu Pro Gly His

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His Pro Pro Ala Lys Asp Ser Lys Thr His Gln Ala Thr Gly Asn Gly

245 250 255

Phe Ser Ser Asn Ser Leu Phe Gly Asp Val Phe Ser Ala Thr Ser Ile

260 265 270

Gln Pro Asn Gln Ala Ser Lys Pro Pro Lys Ser Ser Ala Ser Ser Val

275 280 285

Ser Ile Ser Ser Ser Pro Asn Pro Val Ser Ser Gly Ser Gln Gln Lys

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Val Lys Ser Asn Ser Ile Asp Ser Leu Gln Asn Met His Ser Lys Gln

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Pro Ala Gly Tyr Gln Tyr Gln Gln Thr Pro Ser Ser Val Lys Gln Tyr

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Gln His Val Pro Leu Gln Thr Ser Asn Ala Ile Pro Gly Gly Ala Gly

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Asn Thr Ala Ser Ser Gln Ser Gln Leu Pro Trp Pro Arg Met Thr Gln

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Ala Asp Val Gln Lys Tyr Ser Lys Val Phe Val Ala Val Asp Thr Asp

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Arg Asp Gly Lys Ile Thr Gly Glu Gln Ala Arg Asn Leu Phe Leu Ser

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Trp Lys Leu Pro Arg Glu Ile Leu Arg Gln Val Trp Asp Leu Ser Asp

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Gln Asp Asn Asp Ser Met Leu Ser Leu Arg Glu Phe Cys Ile Ser Leu

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Tyr Leu Met Glu Arg Tyr Arg Glu Gly Arg Pro Pro Pro Pro Val Leu

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Pro Thr Ser Ile Met Leu Asp Glu Ala Met Ala Thr Ser Gly Gln Pro

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Thr Ala Val His Ser Gly Ala Ala Trp Arg His Thr Pro Gly Ile Pro

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Gln Pro Gln Gly Thr Lys Gly Thr His Gln Ala Ala Pro Gly Ser Phe

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Gly Lys Pro Pro Arg Pro Val Pro Ile Ser Gln Pro Asp Glu Ala Arg

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Gln Pro Thr Gln Gln Lys Pro Lys Pro Lys Val Pro Val Leu Glu Lys

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His Leu Val Glu Gln Leu Ser Ser Glu Glu Gln Asp Ser Leu Asn Ser

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Lys Phe Gln Glu Ala Thr Glu Ala Glu Lys Lys Val Ala Glu Leu Glu

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Lys Glu Ile Met Asp Ala Lys Glu Lys Ile Gln Phe Phe His Ala Lys

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Met Gln Glu Leu Ile Leu Tyr Lys Ser Arg Cys Asp Asn Arg Leu Asn

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Glu Ile Thr Asp Arg Thr Ser Ala Asp Lys Lys Glu Val Glu Leu Phe

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Ala Lys Lys Tyr Glu Glu Lys Tyr Lys Gln Thr Gly Asp Val Ala Ser

610 615 620

Lys Leu Thr Ile Gln Glu Ala Thr Phe Arg Asp Ile Gln Glu Lys Lys

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Met Glu Leu Tyr Gln Ala Ile Val Lys Met Asp Gln Asp Gly Asn Ala

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Asp Ser Thr Lys Asp Arg Ala Asn His Ile Gln Lys Asp Leu Glu Glu

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Leu Ile Lys Ser Leu Asn Glu Arg Cys Lys Thr Tyr Gly Leu Arg Ala

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Lys Pro Thr Ser Leu Leu Glu Leu Pro Phe Gly Trp Gln Pro Gly Ile

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Gln Glu Gly Ala Ala Asp Trp Asp Gly Asp Trp Asp Lys Phe Glu Asp

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Glu Gly Phe Thr Ser Leu Lys Glu Leu Thr Leu Asp Val Gln Asn Val

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Ile Ala Pro Ser Lys Thr Lys Ser Ser Leu Ile Arg Glu Lys Val Ser

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Ser Gly Asp Ala Arg Glu Thr Gly Lys Ser Arg Leu Asp Ala Asp Val

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Gly Ala Glu Asn Leu Ser Ser Pro Val Lys Ser Thr Val Val Asp Glu

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Val Thr Ser Val His Ser Asp Asp Gln Arg Ala Arg Ser Pro Pro Glu

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Ser Pro Ser Lys Ser Asn Ala Phe Asp Ser Pro Ser Lys Glu Leu Arg

805 810 815

Glu Phe Gln Pro Arg Lys Glu Phe Asn Phe Asp Gly Ser Pro His Ala

820 825 830

Met Gln Gly Asp His Gly Gly Ala Glu Ser Val Phe Ser Ser Asp Lys

835 840 845

Gly Phe Asp Glu Ser Gly Trp Gly Thr Phe Asp Thr Asn Tyr Asp Ser

850 855 860

Asp Ala Ala Trp Asp Phe Asn Arg Val Ala Ser Lys Asn Ala Asp Asn

865 870 875 880

Glu Thr Gln Lys Glu Asn Pro Leu Phe Gly Phe Asn Asp Trp Gly Leu

885 890 895

Ala Pro Ile Lys Thr Gly Ser Lys Tyr Ala Val Asp Thr Val Pro Lys

900 905 910

Leu Gly Pro Ser Phe Asp Ser Val Pro Ser Thr Pro Ser Tyr Asn Thr

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Gly Ala Pro Ser Ala Gly Asp Val Leu Pro Lys Gln Ser Leu Phe Phe

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Asp Ser Val Pro Ser Thr Pro Ser Tyr Asn Ala Pro Thr Gln Ala Gly

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Asp Lys Phe Ser Lys Gln Met Pro Phe Phe Asp Ser Val Pro Ser Thr

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Pro Ser Tyr Asn Ala Gly Phe Thr Gln Ala Gly Asp Thr Phe Ser Lys

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Gln Ser Ser Phe Phe Asp Ser Val Pro Ser Thr Pro Asn Tyr Asn Ala

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Gly Phe Thr Gln Ala Gly Asp Thr Phe Ser Lys Gln Gly Ser Phe

1010 1015 1020

Phe Asp Ser Val Pro Ser Thr Pro Ser Tyr Asn Thr Gly Phe Ser

1025 1030 1035

Tyr Thr Glu Asn Ala Phe Ser Lys Gln Ser Pro Phe Phe Asp Ser

1040 1045 1050

Val Pro Ser Thr Pro Ala Tyr Ser Ser Asn Leu His Ala Asp Asp

1055 1060 1065

Met Phe Gln Arg Lys Ser Ser Phe Ala Asp Ser Val Pro Ser Thr

1070 1075 1080

Pro Met Tyr Gly Ser Thr Asn Ser Pro Arg Arg Phe Ser Glu Gly

1085 1090 1095

Pro Glu Glu Phe Ser Arg Asp Phe Ser Arg Phe Asp Ser Phe Ser

1100 1105 1110

Ser His Asp Gly Ser Asn Leu Phe Ala Pro Asp Ala Ser Phe Ser

1115 1120 1125

Arg Phe Asp Ser Met Arg Ser Thr Lys Asp Ser Glu Phe Asp His

1130 1135 1140

Gly Leu Phe Pro Pro His Asp Ser Leu Ala Arg Phe Asp Ser Phe

1145 1150 1155

Arg Ser Thr Ala Asp Ser Glu Tyr Thr Phe Gly Asn Pro Pro Pro

1160 1165 1170

Arg Asp Ser Phe Ala Arg Phe Asp Ser Phe Arg Ser Thr Lys Asp

1175 1180 1185

Ser Glu Tyr Gly His Gly Phe Ala Ser Phe Asp Asp Ala Asp Pro

1190 1195 1200

Phe Gly Ser Ser Gly Pro Phe Lys Thr Ser Phe Glu Ser Glu Thr

1205 1210 1215

Pro Arg Arg Asp Ser Ser Gly Pro Phe Lys Thr Ser Phe Glu Ser

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Glu Thr Pro Arg Arg Asp Ser Asp Ser Trp Lys Ala Phe

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<210> 2

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<212> DNA

<213> Ipomoea batatas

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atggctgctc cgaatatgga tcaattcgag gcgtattttc ggagagctga tttggatcag 60

gacggccgga tcagtggtgc cgaggctgtt tccttcctcc aaggctccaa tttaccgaaa 120

caagtgcttg ctcagatatg gatgtatgcg gatcaaagtc aaacaggatt cctaagtcgt 180

caagattttt ataatgcttt aaaactagtt actgtggctc aaagtaaacg agaattgact 240

ccagatatcg ttaaggctgc tttgtttggt ccagcttcag cgaaaattcc agcaccccag 300

ataaaccttg cagccatacc aggtcctcaa ccaaataata tgtcaaattc tccagttcct 360

tcagttggtg ctggtgttcc agcagcaggt ccaagtagtg gaaccagagg tcatcaggtc 420

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ttccagtctc atccggctgt ttcaggtcca ggtgtgccag tgggatccac tatgacttct 540

tcaaattctc ctatttcccc tgatatgaat ggtggtagga ctagtggctc ccaaccaggg 600

gtaacacccc aattgtctta cagaggtatc agtcctaaga gtcatgatgg atttggtctt 660

gttgcctctg gatcaacccc tccacaatca aaaccacaag atgcagcttt accgggtcac 720

catcctccag ctaaggactc taaaactcat caagctacag gaaatggatt ttcttcgaac 780

tctctttttg gagatgtttt ctctgcgaca tctatccagc caaaccaagc ttctaagcca 840

cctaaatctt ctgctagtag tgtatcaatt tcatcatctc caaaccctgt atcttctggt 900

tctcaacaaa aggttaagtc gaactcaatt gattctttgc aaaacatgca ctctaagcaa 960

ccagcaggtt atcagtatca gcaaactcca tcaagtgtga aacaatacca acatgtcccg 1020

ctgcaaactt ccaatgcaat tcctggtgga gctggaaata ccgcatctag ccagtcacag 1080

cttccttggc caagaatgac ccaggctgat gttcagaagt atagtaaagt ctttgtagca 1140

gtagacacag acagggatgg aaaaatcacg ggtgaacaag cacgcaactt gtttttgagc 1200

tggaaacttc ctcgagagat tttacggcag gtatgggact tatctgatca agataatgac 1260

agcatgcttt ctctgaggga attttgtatt tctctctact tgatggaacg atacagggaa 1320

ggtcgccctc ctcctccagt cctaccaact agtattatgc ttgatgaagc aatggctact 1380

tctggtcaac ctactgcagt acattctgga gcagcttgga gacatactcc aggtatacca 1440

caaccacagg gaacaaaagg tacacatcaa gcagctcctg gttcatttgg gaagccacct 1500

cgtccagttc caatttctca gcctgatgaa gctaggcaac ctactcaaca aaagcctaaa 1560

cctaaagttc cggtgcttga gaagcatctt gttgaacaac ttagttcaga agaacaagat 1620

tctctgaact caaagttcca ggaagcaact gaagcagaaa agaaggttgc agaactggag 1680

aaggaaataa tggatgcaaa ggaaaaaatt cagttctttc atgcaaagat gcaagagctt 1740

attttatata aaagccgatg tgacaataga ctaaatgaga tcactgatag gacttctgct 1800

gacaaaaaag aggttgaatt atttgcaaag aaatatgaag agaagtacaa acagactgga 1860

gatgtagcat ctaagttgac aattcaggag gccacttttc gtgatattca ggagaaaaaa 1920

atggagcttt atcaagcaat tgtcaaaatg gatcaagatg gaaatgctga tagtactaag 1980

gatcgtgcta atcacattca aaaggatctt gaggagctta taaaatcatt gaatgaacgc 2040

tgcaaaactt atggtctacg tgccaaacca acttcactgc tggagcttcc atttggttgg 2100

caacctggca tccaagaagg agctgcggat tgggatggtg attgggataa gtttgaagat 2160

gaaggtttca catctttgaa agaactcacc cttgatgtgc aaaatgtgat agctccatct 2220

aagacgaaat cttccctaat tcgggagaaa gtatcttctg gtgatgccag agagacagga 2280

aaatcacgct tggatgctga tgttggggct gagaatttgt caagcccggt aaaaagtaca 2340

gtggtggatg aagtgacatc tgtccatagt gatgaccaaa gagcaagaag tcctcctgaa 2400

agcccgtcaa agagcaatgc atttgatagc ccctctaagg aactgcgtga atttcaacct 2460

aggaaggaat tcaattttga tggttcaccc catgcaatgc aaggtgacca tgggggtgct 2520

gaatccgtgt tctccagtga caaaggtttt gatgaatcag gttggggtac atttgataca 2580

aactatgatt cagatgctgc atgggacttc aatcgtgttg ctagtaagaa tgctgataat 2640

gagacacaga aggagaatcc tttatttggt ttcaatgact ggggtcttgc tcctataaaa 2700

actggatcca aatatgcagt cgacacagtt cccaagctgg gtccatcttt tgactccgtt 2760

cctagtaccc caagttacaa tacaggtgct ccatcagcag gtgatgtatt accaaaacaa 2820

agtctattct ttgattcggt tcctagtact ccaagttaca atgcccccac acaagctgga 2880

gacaagtttt caaaacaaat gccattcttt gattctgttc ctagtactcc aagttacaat 2940

gctggattca cacaagctgg tgacacattc tcaaagcaaa gctcattctt tgattctgtt 3000

cccagtactc caaattacaa tgctggattc acacaagctg gtgacacatt ctcaaagcaa 3060

ggctcattct tcgattctgt tcctagtact cctagttata acactgggtt ctcctacaca 3120

gaaaatgcat tctcaaagca aagtcctttc tttgattctg tgccaagcac gcctgcatat 3180

agttccaacc tacatgcaga tgacatgttt cagaggaaga gctccttcgc cgattctgtc 3240

ccaagtactc caatgtatgg ctctaccaac tccccaagaa ggtttagtga agggcccgaa 3300

gagttctcaa gagacttctc tagatttgat tccttcagct cgcatgatgg cagtaacctc 3360

tttgcacctg atgcatcctt ctcgagattt gactccatgc gcagcaccaa ggactctgaa 3420

tttgatcacg gtttgtttcc accacatgac tctctagcaa gatttgattc cttccgcagc 3480

actgcagact ctgaatatac ctttgggaac cctcctcccc gagactcatt tgcgaggttt 3540

gattccttcc gaagcactaa ggactcagaa tacggtcatg gttttgcatc ttttgatgat 3600

gccgatccat ttggatcaag tgggccattt aagacatcat ttgagagcga gacaccacgg 3660

agagattcaa gtgggccatt taagacatca ttcgagagcg agacaccacg gagagactct 3720

gatagctgga aagcgtttta a 3741

Claims

1. The application of the protein is (c1), (c2), (c3), (c4), (c5) or (c 6):

(c1) regulating and controlling the plant height of the plant;

(c2) promoting plant dwarfing;

(c3) the plant height is promoted to be reduced;

(c4) promoting the plant root length to be shortened;

(c5) reducing the auxin content;

(c6) reducing gibberellin content in plants;

the protein is (a1) or (a 2):

(a1) protein shown as a sequence 1 in a sequence table;

(a2) is derived from protein which has more than 98 percent of identity with the sweet potato (a1) and has the function of regulating and controlling the plant height of the plants.

2. The application of the nucleic acid molecule for coding the protein is (d1), (d2), (d3), (d4), (d5) or (d 6):

(d1) cultivating transgenic plants with changed plant height;

(d2) cultivating a dwarfing transgenic plant;

(d3) cultivating transgenic plants with reduced plant height;

(d4) cultivating transgenic plants with shortened root length;

(d5) cultivating transgenic plants having reduced auxin content;

(d6) cultivating transgenic plants having reduced gibberellin content;

the protein is (a1) or (a 2):

(a1) protein shown as a sequence 1 in a sequence table;

3. Use according to claim 2, characterized in that: the nucleic acid molecule is (b1) or (b2) as follows:

(b2) a DNA molecule derived from sweetpotato and having 95% or more identity to (b1) and encoding the protein of claim 1.

4. A method of making a transgenic plant comprising the steps of: introducing nucleic acid molecules encoding proteins into a starting plant to obtain a transgenic plant with reduced plant height and/or shortened root length;

the protein is (a1) or (a 2):

(a1) protein shown as a sequence 1 in a sequence table;

5. A method of plant breeding comprising the steps of: increasing the content and/or activity of protein in the target plant, thereby reducing the plant height and/or root length of the plant;

the protein is (a1) or (a 2):

(a1) protein shown as a sequence 1 in a sequence table;

6. A method of making a transgenic plant comprising the steps of: introducing nucleic acid molecules encoding proteins into the starting plant to obtain a transgenic plant with reduced auxin and/or gibberellin content;

the protein is (a1) or (a 2):

(a1) protein shown as a sequence 1 in a sequence table;

7. A method of plant breeding comprising the steps of: increasing the content and/or activity of protein in the target plant, thereby reducing the content of auxin and/or gibberellin in the plant;

the protein is (a1) or (a 2):

(a1) protein shown as a sequence 1 in a sequence table;

8. The method of claim 4 or 6, wherein: the nucleic acid molecule is (b1) or (b2) as follows: