A protein ppg1 for regulating poplar growth and development and an encoding gene and application thereof

Abstract

The application discloses a protein PPG1 for regulating growth and development of poplar, a coding gene and application thereof, belongs to the technical field of biology, the amino acid sequence of the protein PPG1 is as shown in SEQ ID NO.2, the gene for coding the PPG1 protein is named as PPG1 gene, and the open reading frame of the PPG1 gene in the cDNA in the poplar is as shown in SEQ ID NO.1 of the sequence listing.The PPG1 gene in the genomic DNA of the poplar is as shown in SEQ ID NO.3 of the sequence listing.The application obtains a PPG1 gene overexpression plant by constructing a PPG1 gene overexpression vector, and the plant growth of the PPG1 gene overexpression strain is better compared with the wild type poplar, and the plant height is significantly increased.It is indicated that the PPG1 gene plays an important regulating function in the growth and development process of the poplar.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biotechnology, and particularly relates to a protein PPG1 for regulating the growth and development of poplar, a coding gene thereof and application. BACKGROUND

[0002] The growth of plants is plastic, and as immobile photosynthetic organisms, plants can adjust their development according to changes in surrounding environmental factors. The ability of plants to regulate their size has important implications for their resistance to extreme climate changes, prevention of pathogen invasion, etc. The growth and development of plants not only determines the overall health status of the plants, but also indirectly affects the resistance of the plants to abiotic stress. Larger plants usually have stronger survival ability, because they can more effectively obtain resources such as light, water and nutrients to support their growth and physiological activities. In addition, larger plants usually have a stronger defense system, including thicker cell walls and more internal defense substances, to resist external environmental pressure.

[0003] Trees are important biological energy sources in the world, and also play a key role in the protection of the ecological environment. Poplar (Populus L.) is one of the main afforestation tree species in northern forestry production in China, and is a component of renewable biological resources and forest carbon sinks, and has important economic and ecological value. Poplar has strong rapid growth, but there are many limiting factors affecting its rapid growth, such as photosynthesis. In order to improve the yield of forestry products to meet the demand of human beings and society for wood resources, the present application finds excellent genes for regulating the growth of trees through molecular biological means combined with other biological techniques, and cultivates excellent varieties through biological engineering means. SUMMARY

[0004] One of the purposes of the present application is to provide a protein PPG1 for regulating the growth and development of poplar and a coding gene thereof.

[0005] The second purpose of the present application is to provide the application of the above-mentioned protein PPG1 and coding gene thereof in promoting the growth and development of poplar.

[0006] In order to achieve the above-mentioned purposes, the technical solutions of the present application are as follows:

[0007] The protein PPG1 for regulating the growth and development of poplar is as follows A1), A2) or A3):

[0008] A1) the amino acid sequence is the protein shown in SEQ ID NO. 2;

[0009] A2) the protein with the same function obtained by substituting, deleting and / or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO. 2 in the sequence listing;

[0010] A3) a fusion protein obtained by connecting a tag at the N-terminal or / and C-terminal of the protein of A1) or A2).

[0011] The protein of A2) above is a protein having 75% or more identity with the amino acid sequence of the protein shown in SEQ ID NO. 2 and having the same function. The 75% or more identity means 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity.

[0012] The protein of A2) above can be artificially synthesized or can be obtained by first synthesizing its encoding gene and then performing biological expression.

[0013] The encoding gene of the protein of A2) above (named PPG1 gene) can be obtained by deleting one or several codons of the amino acid residues in the DNA sequence shown in SEQ ID NO. 1, and / or performing one or several base pair missense mutations, and / or connecting the encoding sequence of the tag shown in the above table at its 5' end and / or 3' end.

[0014] The present application also provides the encoding sequence of the above protein PPG1, and the encoding sequence is shown in SEQ ID NO. 1. The DNA molecule shown in SEQ ID NO. 1 encodes the protein PPG1 shown in SEQ ID NO. 2.

[0015] Further defined, the encoding sequence also includes any one of the following:

[0016] (1) the encoding sequence is a cDNA molecule or a DNA molecule of SEQ ID NO. 1;

[0017] (2) the cDNA molecule or the DNA molecule shown in SEQ ID NO. 1 in the sequence listing;

[0018] (3) the DNA molecule shown in SEQ ID NO. 3 in the sequence listing;

[0019] (4) a cDNA molecule or a DNA molecule having 75% or more identity with the encoding sequence shown and encoding the protein.

[0020] (5) a cDNA molecule or a genomic DNA molecule hybridizing with the encoding sequence defined in (1) or (2) or (3) or (4) above under stringent conditions and encoding the protein.

[0021] Further defined, the encoding sequence can be DNA, such as cDNA, genomic DNA or recombinant DNA, or can be RNA, such as mRNA or hnRNA, etc.

[0022] The term "identity" as used herein refers to sequence similarity with a native nucleic acid sequence. "Identity" includes nucleotide sequences having 75% or more, or 85% or more, or 90% or more, or 95% or more identity with the nucleotide sequence encoding the protein consisting of the amino acid sequence shown in SEQ ID NO. 2 of the present application. Identity can be assessed by eye or by computer software. Using computer software, identity between two or more sequences can be expressed as a percentage (%) which can be used to assess identity between related sequences.

[0023] The 75% or more identity described above can be 80%, 85%, 90% or 95% or more identity.

[0024] The present application also provides biological materials related to the above-mentioned coding sequence, and the biological materials are any one of the following:

[0025] B1) a nucleic acid molecule encoding PPG1 protein;

[0026] B2) a gene expression cassette containing the sequence or partial sequence of B1);

[0027] B3) a recombinant vector containing the expression cassette of B1);

[0028] B4) a recombinant microorganism containing the nucleic acid molecule of B1);

[0029] B5) a recombinant microorganism containing the expression cassette of B1);

[0030] B6) a recombinant microorganism containing the recombinant vector of B2);

[0031] A7) a recombinant microorganism containing the recombinant vector of B3);

[0032] A8) a transgenic plant cell line containing the recombinant vector of B2);

[0033] A9) a transgenic plant cell line containing the recombinant vector of B3).

[0034] In addition, those skilled in the art can easily mutate the PPG1 gene described in the present application using known methods, such as methods of directed evolution and point mutation. Those nucleotides artificially modified, having 75% or more identity with the nucleotide sequence encoding the protein PPG1, as long as they encode the same protein and have the same function, are derived from the nucleotide sequence of the present application and are equivalent to the sequence of the present application. Primer pairs amplifying the entire length of the coding sequence encoding the protein PPG1 or fragments thereof also fall within the scope of the present application.

[0035] The most important purpose of the present application is to protect the application of the above-mentioned protein PPG1, the PPG1 gene encoding the protein PPG1 and the biological material containing the above-mentioned coding sequence in promoting the growth and development of poplar.

[0036] The promotion of the growth and development of poplar specifically shows that, compared with the wild type, the plant height of the PPG1 gene overexpression strain is higher, the photosynthesis is stronger, the biomass is larger, and the growth is better.

[0037] The present application also protects a method for promoting the growth and development of poplar, which comprises:

[0038] (1) increasing the activity of the protein PPG1 in poplar or increasing the content of the protein PPG1 in poplar;

[0039] (2) promoting the expression of the PPG1 gene in poplar;

[0040] Among them, the way to promote the expression of the PPG1 gene in poplar can be overexpression or superexpression.

[0041] In the above-mentioned method for promoting the growth and development of poplar, the PPG1 gene overexpression plant is obtained by overexpression of the PPG1 gene in the plant genome, and the homozygous PPG1 gene overexpression plant is obtained from the selfed offspring of the PPG1 gene overexpression plant; the homozygous PPG1 gene overexpression plant is a plant that grows and develops faster. The selfed offspring can be the offspring of the first selfed generation, the offspring of the second selfed generation, the offspring of the third selfed generation, etc., until the homozygous PPG1 gene overexpression plant is obtained. The selfed offspring of the homozygous PPG1 gene overexpression plant is a relatively high-speed growth strain.

[0042] The above-mentioned method for promoting the growth and development of poplar is also applicable to other receptor plants having homologous genes with poplar, and there is no particular limitation on the receptor plants suitable for the present application, which not only includes poplar, but also other plants with high homology, as long as they are suitable for gene transformation operation, such as various crops, ornamental plants or forestry plants, etc. The plants can be (not limited to) dicotyledonous plants, monocotyledonous plants, woody plants, rosaceous plants, rosaceous plants, peach, cruciferous plants, arabidopsis plants, arabidopsis, etc.

[0043] The "plant" in the present application includes the whole plant, the parent and offspring plants and different parts of the plant, including seeds, fruits, buds, stems, leaves, roots (including tubers), flowers, tissues and organs, and the gene or nucleic acid of interest is present in these different parts. The "plant" mentioned herein also includes plant cells, suspension cultures, callus, embryos, meristematic regions, gametophytes, sporophytes, pollen and microspores, and each of the foregoing objects also contains the gene / nucleic acid of interest.

[0044] The present invention includes any plant cell, or any plant obtained or obtainable by the methods therein, and all plant parts and propagules thereof. The present patent also encompasses transfected cells, tissues, organs or whole plants obtained by any of the aforementioned methods. The only requirement is that the progeny exhibit the same genotypic or phenotypic characteristics as obtained using the methods of the present patent.

[0045] The present invention also extends to harvestable parts of the plants as described above, but not limited to seeds, leaves, fruits, flowers, stems, roots, rhizomes, tubers and corms. It further relates to other derivatives of the plants after harvesting, such as dried pellets or powders, oils, fats and fatty acids, starches or proteins. The present invention also relates to foodstuffs or food additives obtained from the related plants.

[0046] Advantages of the present invention:

[0047] The present invention is advantageous in that a new protein PPG1 regulating the growth and development of plants was discovered from poplar during the research, and it was proved by experiments that the plant height of the PPG1 gene overexpression strain was higher, the photosynthesis was stronger, the biomass was larger, and the growth was better compared with the wild type, that is, the development of the PPG1 gene overexpression strain was significantly better than that of the wild type material, which indicated that the PPG1 gene played an important regulatory function in the growth and development of poplar, and had important guiding significance for the genetic improvement of poplar. BRIEF DESCRIPTION OF DRAWINGS

[0048] Figure 1 is the result of yeast double hybrid experiment.

[0049] Figure 2 is an example of a cell under laser scanning confocal microscope.

[0050] Figure 3 is an example of the T4 generation plants of wild type (WT) and PPG1 gene overexpression strain (35S-PPG1-T4).

[0051] Figure 4 is the analysis of fresh weight and plant height of the T4 generation plants of wild type and PPG1 gene overexpression strain; in the figure: A figure represents the fresh weight of four groups of T4 generation PPG1 gene overexpression strain and wild type plants after being cultured for the same number of days under the same conditions, and B figure represents the plant height of four groups of T4 generation PPG1 gene overexpression strain and wild type plants after being cultured for the same number of days under the same conditions.

[0052] Figure 5Figure 4 is an analysis of ETR, Fv / Fm and NPQ of T4 generation plants of wild type and PPG1 gene overexpression line; in the figure, A represents the change of electron transport rate (ETR) of T4 generation PPG1 gene overexpression line and wild type plant with gradually increasing light intensity, B represents the change of Fv / Fm of T4 generation PPG1 gene overexpression line and wild type plant with time, and C represents the change of non-photochemical quenching (NPQ) of T4 generation PPG1 gene overexpression line and wild type plant with time. DETAILED DESCRIPTION

[0053] The advantages and features of the present application will become apparent from the descriptions given to the specific embodiments, it being understood that the following embodiments are intended to be illustrative only and the present application can be carried out in a variety of ways, not all of which can be described here. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Methods and materials are described herein for use in the present application; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0054] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Methods and materials are described herein for use in the present application; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0055] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Methods and materials are described herein for use in the present application; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0056] As used herein, the terms "nucleic acid", "nucleic acid sequence", "nucleotide", "nucleic acid molecule" or "polynucleotide" are meant to include isolated DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., messenger RNA), naturally occurring types, mutated types, synthetic DNA or RNA molecules, DNA or RNA molecules composed of nucleotide analogs, single- or double-stranded structures. These nucleic acids or polynucleotides include, but are not limited to, coding sequences of genes, antisense sequences, and regulatory sequences of non-coding regions. These terms include a gene. A "gene" or "gene sequence" is used broadly to refer to a functional DNA nucleic acid sequence. Thus, a gene can include both exons and introns of a genomic sequence, and / or the coding sequence of a cDNA, and / or regulatory sequences of eDNA and its regulatory sequences. In particular embodiments, such as with respect to isolated nucleic acid sequences, it is preferred that they be cDNA.

[0057] Unless otherwise indicated, the first nucleotide of each nucleotide sequence in the sequence listing is the 5' end nucleotide of the corresponding DNA / RNA, and the last nucleotide is the 3' end nucleotide of the corresponding DNA / RNA.

[0058] The inventors of the present application found a new protein regulating plant growth and development from poplar, named PPG1 protein. The amino acid sequence of PPG1 protein in poplar is shown as SEQ ID NO. 2 in the sequence listing. The gene encoding PPG1 protein is named PPG1 gene, and the gene number in NCBI database is LOC7454940, and the gene sequence ID is XM 002309035.4. The open reading frame of PPG1 gene in cDNA in poplar is shown as SEQ ID NO. 1 in the sequence listing. The PPG1 gene in genomic DNA of poplar is shown as SEQ ID NO. 3 in the sequence listing.

[0059] Example 1: Isolation of PPG1 gene

[0060] The protein PPG1 interacting with poplar photosystem II protein D1 was obtained by using yeast two-hybrid screening library. The sequence of poplar photosystem II protein D1 is shown as SEQ ID NO. 4, and the sequence of the encoding gene is shown as SEQ ID NO. 5.

[0061] The specific method is as follows:

[0062] 1. The D1 gene shown as SEQ ID NO. 5 is amplified by PCR, and the gene and pGBKT7 vector are double-digested by restriction endonuclease EcoRI and SalI, the enzyme digestion product is recovered, and the bait vector D1-pGBKT7 (BD) is obtained by ligation.

[0063] 2. The leaves of the tissue culture seedlings of 84K poplar are cut and ground under liquid nitrogen environment for standby. Total RNA is extracted by using Tiangen total RNA extraction kit. The specific operation steps refer to the instruction manual of total RNA extraction kit (RNAEasy Fast Plant Tissue Kit) of Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0064] 3. cDNA synthesis

[0065] (1) The reverse transcription process adopts Beijing Quanshi Gold Bio Trans Script One-Step gDNA Removal and cDNA Synthesis Super Mix kit, and the reverse transcription mixed solution is configured according to the following system:

[0066] Total RNA / mRNA 50 ng-5 μg;

[0067] Anchored oligo(dT) 18 primer 1 μL;

[0068] 2 x TS Reaction Mix 10 μL;

[0069] Trans Script RT / RI Enzyme Mix 1 μL;

[0070] g DNA Remover 1 μL;

[0071] RNase-free Water to 20 μL.

[0072] (2) Mix gently and incubate at 42°C for 30 min;

[0073] (3) Heat at 85°C for 5 s to inactivate Trans Script RT / RI and g DNA Remover.

[0074] 4. The obtained cDNA was ligated to pGADT7 vector to construct a library, and the library and the bait vector D1-pGBKT7 (BD) were transformed into Y2H Gold together, and the bait vector D1-pGBKT7 (BD) and the empty pGADT7 vector were also transformed into Y2H Gold together to verify whether the bait vector has self-activation, and a positive control and a negative control were prepared.

[0075] 5. The yeast cells transformed with the library were coated on a culture medium containing X-Gal to screen positive clones activating the LacZ reporter gene. The results are shown in Figure 1 The yeast clones growing on the screening medium were picked up for sequencing of the prey protein gene to obtain the corresponding PPG1 gene.

[0076] Example 2 Cell sub-localization of PPG1 protein

[0077] 1. The double-stranded DNA molecule shown in SEQ ID NO. 1 of the sequence listing was used as a template, and a primer pair composed of PPG1-GFPS and PPG1-GFPA was used for PCR amplification, and the PCR amplification product was recovered.

[0078] PPG1-GFPS:

[0079] 5'-CCGGAATTCATGGCAACTTTAACTCCTATTCAGACTT-3';

[0080] PPG1-GFPFPA:

[0081] 5'-CCGGGTACCTTTATCAATTTGGTTGAGCTTGCCAAG-3'.

[0082] 2. Take the amplified product obtained in step 1, and perform double enzyme digestion with restriction enzymes EcoRI and KpnI, and recover the enzyme digestion product.

[0083] 3. Take PBSK vector (Biovector 105802, Sincere Biotechnology Co., Ltd.), and perform double enzyme digestion with restriction enzymes EcoRI and KpnI, and recover the vector skeleton.

[0084] 4. Connect the enzyme digestion product obtained in step 2 and the vector skeleton obtained in step 3 to obtain recombinant plasmid PBSK-PPG1.

[0085] 5. Take the leaves of wild-type poplar germinated for 3-4 weeks, cut the middle part of the leaves into 0.5-1 mm strips, and place the strips in an enzyme digestion solution for dark digestion for 3-4 h, centrifuge, and collect the protoplasts.

[0086] 6. Transform the protoplasts with the recombinant plasmid PBSK-PPG1 by using the PEG method, and culture under light overnight.

[0087] 7. Observe the expression of GFP in mesophyll cells by using a laser scanning confocal microscope (LSM510 META, Zeiss). The objective magnification is 40 times, the excitation light wavelength is 488 nm, the band pass BP505-530 nm, and the long pass LP560 nm.

[0088] The results of cells under the laser scanning confocal microscope are shown in Figure 2 , which shows that the PPG1-GFP signal completely overlaps with the chlorophyll autofluorescence signal, indicating that the PPG1 protein is located in the chloroplast.

[0089] Example 3 Functional identification of PPG1 gene

[0090] I. Construction of PPG1 gene overexpression vector

[0091] 1. Amplification of target gene PPG1

[0092] Synthesize pCAMBIA1300-35S-PPG1-F and pCAMBIA1300-35S-PPG1-R, respectively, and PCR amplify the PPG1 gene shown in SEQ ID NO. 1.

[0093] pCAMBIA1300-35S-PPG1-F:

[0094] 5'-CCGggtaccATGGCAACTTTAACTCCTATTCAGACTT-3';

[0095] pCAMBIA1300-35S-PPG1-R:

[0096] 5'-GGCggatccTTTATCAATTTGGTTGAGCTTGCCAAG-3'.

[0097] 2. Using restriction enzymes Kpn I and BamH I, the PPG1 fragment obtained in the previous step is double-digested with pCAMBIA1300-35S vector, and the resulting pCAMBIA1300-35S-PPG1 recombinant vector is sequenced and verified.

[0098] The sequencing results show that the pCAMBIA1300-35S-PPG1 recombinant vector is a fragment between the Kpn I and BamH I enzyme sites of the pCAMBIA1300-35S vector, which is replaced by the PPG1 gene, and the other sequences of the pCAMBIA1300-35S vector remain unchanged. The obtained pCAMBIA1300-35S-PPG1 recombinant vector can overexpress PPG1 protein.

[0099] II. Obtain PPG1 gene overexpression plants

[0100] 1. The PPG1 gene overexpression vector constructed in step one is introduced into Agrobacterium GV3101 to obtain recombinant Agrobacterium.

[0101] 2. The 1-3 month old plants are cultured, and the leaves with uniform leaf color and no any abnormalities on the leaf surface are selected. The edges of the leaves are removed, and the wounds are evenly cut in the middle of the leaves. The recombinant Agrobacterium obtained in step 1 is inoculated into the infection liquid, and the concentration is adjusted to OD600=0.3-0.4. The prepared leaves are placed in the bacterial liquid, and shaken constantly during the period. After 15 min of infection, the leaves are taken out and torn into small pieces, and inoculated into co-culture medium for dark culture at 25°C for 2d.

[0102] 3. The co-cultured leaves are inoculated on the induction and screening medium, and continue to be dark cultured for about two weeks. After the callus grows out, it is subcultured to new screening medium, and subcultured once every two weeks. Generally, the induction and subculture are performed for a maximum of 3 times, and the longest period of the whole stage is 45d. The induced callus is inoculated on the differentiation medium, and subcultured about once every three weeks. During this period, the callus will turn green and hard. Under normal circumstances, this stage is about two months. The small seedlings growing to 1-3 cm are taken down and inoculated on the rooting screening medium, and can root in about 7-15d. The normally growing plants are transplanted into nutrient soil for culture, which are T1 generation plants.

[0103] 4. PPG1 gene overexpression plants are screened from T1 generation plants.

[0104] Specific steps: take the leaves of the plants, extract the genomic DNA by CTAB method, use the primer pair composed of primer F and primer R for PCR amplification, then recover the PCR amplification product and sequence, and screen the PPG1 gene overexpression plants according to the sequencing results.

[0105] Primer F: 5'-ATGGCAACTTTAACTCCTATTCAGACTT-3';

[0106] Primer R: 5'-TTTATCAATTTGGTTGAGCTTGCCAAG-3'.

[0107] 5, self-cross the PPG1 gene overexpression plants obtained in step 4 and harvest the seeds.

[0108] 6, sow the seeds harvested in step 5 in solid 1 / 2MS medium containing 50 mg / L hygromycin, and after 10 days of culture, transplant the normally growing plants to the nutrient soil for culture, which are T2 generation plants.

[0109] 7, screen the PPG1 gene overexpression plants from the T2 generation plants (the specific steps are the same as step 4).

[0110] 8, self-cross the PPG1 gene overexpression plants obtained in step 7 and harvest the seeds.

[0111] 9, sow the seeds harvested in step 8 in the nutrient soil and culture them into plants, which are T3 generation plants.

[0112] 10, identify the T3 generation plants (the specific steps are the same as step 4), and find that the T3 generation plants obtained by self-crossing a certain T2 generation plant are all the same homozygous PPG1 overexpression plants, the T2 generation plant is named as PPG1 overexpression plant, and the offspring of the PPG1 gene overexpression plant is named as PPG1 gene overexpression line.

[0113] 11, self-cross the T3 generation plants of the PPG1 gene overexpression line and harvest the seeds, sow the harvested seeds in the nutrient soil and culture them into plants, which are T4 generation plants.

[0114] 12, self-cross the T4 generation plants of the PPG1 gene overexpression line and harvest the seeds, sow the harvested seeds in the nutrient soil and culture them into plants, which are T5 generation plants.

[0115] The PPG1 gene overexpression plant, the T3 generation plants of the PPG1 gene overexpression line, the T4 generation plants of the PPG1 gene overexpression line, the T5 generation plants of the PPG1 gene overexpression line all have the following phenotypes: in the 1 / 2MS medium with suitable conditions, compared with the wild type poplar, the plants of the PPG1 gene overexpression line grow better and have significantly increased plant height. s ​

[0116] III. Phenotype and physiological identification of poplar

[0117] Poplar to be tested: T4 generation plants of PPG1 gene overexpression line (35S-PPG1-T4) and wild type (WT) poplar.

[0118] For phenotype determination of poplar seedlings, seeds of T4 generation plants of PPG1 gene overexpression line and wild type plants were surface sterilized, placed on 1 / 2MS plates and kept at 4°C for 2 days. Then the plates were transferred to suitable conditions for culture.

[0119] Exemplary photos of the phenotype of T4 generation plants of PPG1 overexpression line and wild type plants germinated for 30 days under suitable conditions are shown in Figure 3 .

[0120] Physiological index determination was performed on T4 generation plants of PPG1 gene overexpression line and wild type plants, including fresh weight and plant height, and the results are shown in A and B of Figure 4 . As shown in the figures, the average fresh weight and plant height of the four groups of T4 generation plants of PPG1 gene overexpression line were significantly higher than those of wild type plants after the same number of days of culture under suitable conditions.

[0121] Photosynthetic parameter determination was performed on T4 generation plants of PPG1 overexpression line and wild type plants, including ETR, Fv / Fm and NPQ, and the results are shown in A, B and C of Figure 5 . As shown in the figures, the ETR, Fv / Fm and NPQ of the four groups of T4 generation plants of PPG1 gene overexpression line were higher than those of wild type plants, which reflects that the PPG1 gene overexpression line has stronger photosynthetic capacity than the wild type plants.

[0122] These results show that under suitable conditions, the T4 generation plants of PPG1 gene overexpression line have higher plant height, stronger photosynthesis and greater biomass and better growth than the wild type plants in the same time, i.e., the T4 generation plants of PPG1 gene overexpression line have higher growth rate and growth amount than the wild type plants in the same time. The above results show that the PPG1 protein and its coding sequence can promote the growth and development of poplar.

[0123] The above-described embodiments are only preferred embodiments of the present application, merely used to explain the present application, and are not intended to limit the scope of the present application. For those skilled in the art, other embodiments can be easily obtained by substitution or change based on the technical content disclosed in the present specification, and therefore, any changes and improvements made on the principles of the present application shall be included in the scope of the patent application.

[0124] The sequence of SEQ ID NO. 1 is as follows:

[0125]

[0126] The sequence of SEQ ID NO. 2 is as follows:

[0127]

[0128] The sequence of SEQ ID NO. 3 is as follows:

[0129]

[0130]

[0131] The sequence of SEQ ID NO. 4 is as follows:

[0132]

[0133] The sequence of SEQ ID NO. 5 is as follows:

[0134]

[0135]

Claims

1. PPG1 The application of genes in promoting the growth and development of poplar trees is characterized by, The specific manifestations of promoting poplar growth and development are as follows: compared with the wild type... PPG1 The gene-overexpressing lines are taller and have stronger photosynthetic activity. PPG1 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

2. Containing the contents of claim 1 PPG1 The application of gene recombinant vectors in promoting poplar growth and development is characterized by, The specific manifestations of promoting poplar growth and development are as follows: compared with the wild type... PPG1 The gene-overexpressing lines are taller and have stronger photosynthetic activity. PPG1 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

3. A method for promoting the growth and development of poplar trees, characterized in that, The method includes: (1) By increasing the content of protein PPG1 in poplar; (2) By promoting the growth of poplar trees PPG1 Gene expression; The specific manifestations of promoting poplar growth and development are as follows: compared with the wild type... PPG1 The gene-overexpressing lines exhibited greater plant height and stronger photosynthetic activity; the amino acid sequence of the protein PPG1 is shown in SEQ ID NO.

2. PPG1 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

4. The method for promoting the growth and development of poplar trees according to claim 3, characterized in that, Promote poplar PPG1 The gene is expressed in the form of overexpression.

5. The method for promoting the growth and development of poplar trees according to claim 4, characterized in that, In plant genomes PPG1 Genes are overexpressed, resulting in PPG1 Gene overexpression plants, from PPG1 Homozygous offspring obtained from self-pollination of gene-overexpressing plants PPG1 Gene overexpression plants; homozygous type PPG1 Plants with overexpressed genes are those that grow and develop faster.

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