Protein PPG1 for regulating and controlling growth and development of poplar as well as coding gene and application of protein PPG1

Abstract

The invention discloses a protein PPG1 for regulating and controlling growth and development of poplars and a coding gene and application thereof, and belongs to the technical field of biology, the amino acid sequence of the protein PPG1 is shown as SEQ ID NO.2, the gene for coding the protein PPG1 is named as PPG1 gene, and the open reading frame of the PPG1 gene in cDNA in the poplars is shown as SEQ ID NO.1 in a sequence table. And the PPG1 gene in the genome DNA of the poplar is as shown in SEQ ID NO.3 in the sequence table. According to the invention, a PPG1 gene overexpression vector is constructed to obtain a PPG1 gene overexpression plant, and compared with a wild poplar, the PPG1 gene overexpression plant has the advantages that the growth vigor of the plant is better, and the plant height is obviously increased. Therefore, the PPG1 gene plays an important regulation and control function in the growth and development process of the poplar.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a protein PPG1 for regulating the growth and development of poplars, a coding gene thereof and an application thereof. Background Art

[0002] Plant growth is plastic. As immobile photosynthetic organisms, plants can adjust their own development according to changes in surrounding environmental factors. The ability of plants to regulate their growth size has an important impact on their ability to withstand extreme climate change and prevent pathogen invasion. Plant growth and development not only determines the overall health of the plant, but also indirectly affects the plant's ability to resist abiotic stress. Larger plants generally have stronger survival abilities because they can more efficiently obtain resources, such as light, water, and nutrients, to support their growth and physiological activities. In addition, larger plants tend to have stronger defense systems, including thicker cell walls and more internal defense substances to resist the pressure of the external environment.

[0003] Trees are an important source of bioenergy in the world and also play a key role in protecting the ecological environment. Poplar (Populus L.) is one of the main afforestation species in forestry production in northern my country. It is a component of renewable biological resources and forest carbon sinks and has important economic and ecological value. Poplars have strong fast-growing properties, but there are many limiting factors that affect their rapid growth, such as photosynthesis. In order to increase the output of forestry products to meet the needs of humans and society for wood resources, this study uses molecular biological methods combined with other biological technologies to find excellent genes that regulate tree growth, and cultivates excellent varieties through bioengineering methods. Summary of the invention

[0004] One of the purposes of the present invention is to provide a protein PPG1 for regulating the growth and development of poplar and its encoding gene.

[0005] The second object of the present invention is to provide the use of the above protein PPG1 and its encoding gene in promoting the growth and development of poplar.

[0006] In order to achieve the above object, the technical solution of the present invention is as follows:

[0007] The protein PPG1 that regulates the growth and development of poplar, the protein is the following A1), A2) or A3):

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

[0009] A2) a protein having the same function as the amino acid sequence shown in SEQ ID NO. 2 in the sequence list, wherein one or more amino acid residues are substituted and / or deleted and / or added;

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

[0011] The protein in A2) 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 is 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity.

[0012] The protein in A2) above can be synthesized artificially, or its encoding gene can be synthesized first and then expressed biologically.

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

[0014] The present invention also provides a coding sequence of the protein PPG1, which 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] It is further defined that the coding sequence also includes any one of the following:

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

[0017] (2) the cDNA molecule or 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 that has 75% or more identity with the coding sequence and encodes the protein.

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

[0021] It is further defined that the coding sequence can be DNA, such as cDNA, genomic DNA or recombinant DNA, or RNA, such as mRNA or hnRNA.

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

[0023] The aforementioned 75% or more identity may be 80%, 85%, 90% or 95% or more identity.

[0024] The present invention also provides a biological material related to the above coding sequence, wherein the biological material is any one of the following materials:

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

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

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

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

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

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

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

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

[0033] A9) A transgenic plant cell line containing the recombinant vector described in B3).

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

[0035] The main purpose of the present invention is to protect the above protein PPG1, the PPG1 gene encoding the protein PPG1 and the application of biological materials containing the above coding sequence in promoting the growth and development of poplars.

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

[0037] The present invention also protects a method for promoting the growth and development of poplars, the method comprising:

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

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

[0040] The method of promoting the expression of the PPG1 gene in poplar can be overexpression or superexpression.

[0041] In the above method for promoting the growth and development of poplar, the PPG1 gene in the plant genome is overexpressed to obtain a PPG1 gene overexpressing plant, and a homozygous PPG1 gene overexpressing plant is obtained from the self-pollinated offspring of the PPG1 gene overexpressing plant; the homozygous PPG1 gene overexpressing plant is a plant with faster growth and development. The self-pollinated offspring can be the offspring of the first generation of self-pollination, the offspring of the second generation of self-pollination, the offspring of the third generation of self-pollination, etc., until a homozygous PPG1 gene overexpressing plant is obtained. The self-pollinated offspring of the homozygous PPG1 gene overexpressing plant is a relatively fast growing strain.

[0042] The above method for promoting the growth and development of poplars is also applicable to other receptor plants with homologous genes to poplars. There is no particular limitation on the receptor plants applicable to the present invention, including not only poplars, but also other plants with high homology, as long as they are suitable for gene transformation operations, such as various crops, flower plants or forestry plants. The plants can be, for example (but not limited to): dicots, monocots, woody plants, Rosales, Rosaceae, Prunus, peach, Cruciferae, Arabidopsis, Arabidopsis, etc.

[0043] The "plant" mentioned in the present invention includes the whole plant, its parent and progeny plants and different parts of the plant, including seeds, fruits, buds, stems, leaves, roots (including tubers), flowers, tissues and organs, and our target gene or nucleic acid is present in these different parts. The "plant" mentioned here also includes plant cells, suspension cultures, callus, embryos, meristem regions, gametophytes, sporophytes, pollen and microspores, and similarly, each of the aforementioned objects contains the target gene / nucleic acid.

[0044] The present invention includes any plant cell, or any plant obtained or obtainable by the method therein, and all plant parts and propagules thereof. This patent also includes 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, and the progeny obtained using the method of this patent have the same characteristics.

[0045] The invention also extends to the harvestable parts of the plants as described above, but not limited to seeds, leaves, fruits, flowers, stems, roots, rhizomes, tubers and bulbs. It also further relates to other derivatives of the plants after harvest, such as dry granules or powders, oils, fats and fatty acids, starch or proteins. The invention also relates to foods or food additives obtained from the relevant plants.

[0046] Advantages of the present invention:

[0047] The present invention discovered a new protein PPG1 that regulates plant growth and development from poplar during the research process, and proved through experiments that, compared with the wild type, the PPG1 gene overexpression strain has a taller plant height, stronger photosynthesis, larger biomass, and better growth. In other words, the development of the PPG1 gene overexpression strain is significantly better than that of the wild type material, indicating that the PPG1 gene plays an important regulatory function in the growth and development of poplar, which has important guiding significance for the genetic improvement of poplar. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 This is the result of yeast two-hybrid experiment.

[0049] Figure 2 The figure is an exemplary photograph of a cell under a laser scanning confocal microscope.

[0050] Figure 3 is the T of wild-type plants (WT) and PPG1 gene overexpression lines 4 Generation plants (35S-PPG1-T 4 ) are shown in Figure 2.

[0051] Figure 4 The fresh weight and plant height of T4 plants of wild-type plants and PPG1 gene overexpression lines were analyzed; Figure A shows that under the same conditions, after the same number of days of cultivation, four groups of T 4 Figure 2 shows the fresh weight of the PPG1 gene overexpression lines and wild-type plants. Figure B shows the fresh weight of the four groups of T plants after being cultured for the same number of days under the same conditions. 4 Plant height of the PPG1 gene overexpression lines and wild-type plants.

[0052] Figure 5The ETR, Fv / Fm and NPQ of T4 plants of wild-type plants and PPG1 gene overexpression lines are analyzed; Figure A shows that as the light intensity gradually increases, T 4 Figure 2. Changes in electron transfer rate (ETR) between the PPG1 overexpression lines and wild-type plants. Figure B shows the changes in electron transfer rate (ETR) between the PPG1 overexpression lines and wild-type plants. 4 The Fv / Fm changes of the PPG1 gene overexpression lines and wild-type plants over time. Figure C shows the T 4 Changes of non-photochemical quenching (NPQ) over time in the first generation PPG1 gene overexpression lines and wild-type plants. DETAILED DESCRIPTION

[0053] The present invention will be further described below in conjunction with specific examples, and the advantages and features of the present invention will become clearer as the description proceeds. However, the specific experimental methods involved in the following examples are all conventional methods or implemented under the conditions recommended by the manufacturer's instructions unless otherwise specified.

[0054] Unless otherwise specified, the technical means used in the examples are conventional means known to those skilled in the art. The test methods in the following examples are conventional methods unless otherwise specified. Unless otherwise specified, the reagents and materials used can all be purchased from the market.

[0055] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[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), natural types, mutant types, synthetic DNA or RNA molecules, DNA or RNA molecules composed of nucleotide analogs, single-stranded or double-stranded structures. These nucleic acids or polynucleotides include gene coding sequences, antisense sequences and regulatory sequences of non-coding regions, but are not limited to these. These terms include a gene. "Gene" or "gene sequence" is widely used to refer to a functional DNA nucleic acid sequence. Therefore, a gene may include introns and exons in a genomic sequence, and / or include coding sequences in cDNA, and / or include cDNA and its regulatory sequences. In specific embodiments, such as with respect to isolated nucleic acid sequences, it is preferably assumed to be cDNA.

[0057] Unless otherwise specified, the first position of each nucleotide sequence in the sequence table is the 5' terminal nucleotide of the corresponding DNA / RNA, and the last position is the 3' terminal nucleotide of the corresponding DNA / RNA.

[0058] The inventors of the present invention discovered a new protein that regulates plant growth and development from poplar during the research process, named PPG1 protein. The amino acid sequence of the PPG1 protein in poplar is shown in SEQ ID NO.2 of the sequence table. The gene encoding the PPG1 protein is named PPG1 gene, the gene number in the NCBI database is LOC7454940, and the gene sequence ID is XM 002309035.4. The open reading frame of the PPG1 gene in the cDNA of poplar is shown in SEQ ID NO.1 of the sequence table. The PPG1 gene in the genomic DNA of poplar is shown in SEQ ID NO.3 of the sequence table.

[0059] Example 1 PPG1 gene isolation

[0060] The poplar photosystem II protein D1 was used to screen the yeast two-hybrid library to obtain the protein PPG1 that interacts with it. The sequence of the poplar photosystem II protein D1 is shown in SEQ ID NO.4, and the sequence of its encoding gene is shown in SEQ ID NO.5.

[0061] The specific method is as follows:

[0062] 1. PCR amplified the D1 gene shown in SEQ ID NO.5, and used restriction endonucleases EcoRI and SalI to double-digest the gene and the pGBKT7 vector, recovered the digestion products, and ligated them to obtain the bait vector D1-pGBKT7 (BD).

[0063] 2. Cut the leaves of the tissue culture seedlings of 84K poplar and grind them in liquid nitrogen for later use. Use Tiangen Plant Total RNA Extraction Kit to extract RNA. For specific operation steps, refer to the instructions of the Plant 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 uses the Beijing Quanshijin Biotechnology Trans Script One-Step gDNA Removal and cDNA Synthesis Super Mix Kit, and the reverse transcription mixture is prepared according to the following system:

[0066] Total RNA / m RNA 50ng-5μg;

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

[0068] 2×TS reaction mix 10 μL;

[0069] TransScript RT / RIEnzyme Mix 1μL;

[0070] g DNARemover 1 μL;

[0071] RNase-free Water to 20 μL.

[0072] (2) Mix gently and use the product for PCR: incubate at 42°C for 30 min;

[0073] (3) Heat at 85°C for 5 seconds to inactivate Trans Script RT / RI and gDNARemover.

[0074] 4. Connect the obtained cDNA to the pGADT7 vector to construct a library, and then transform the library and the bait vector D1-pGBKT7 (BD) into Y2HGold together. Also transform the bait vector D1-pGBKT7 (BD) and the pGADT7 empty vector into Y2HGold together to verify whether the bait vector is self-activated. At the same time, perform positive and negative controls.

[0075] 5. Spread the yeast cells transformed with the library on a medium containing X-Gal to screen out positive clones that activate the LacZ reporter gene. Figure 1 As shown, yeast clones growing on the screening medium were picked, and the prey protein gene was sequenced to obtain the corresponding PPG1 gene.

[0076] Example 2 Cellular sublocalization of PPG1 protein

[0077] 1. Using the double-stranded DNA molecule shown in SEQ ID NO.1 in the sequence table as a template, PCR amplification was performed using a primer pair consisting of PPG1-GFPS and PPG1-GFPA, 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, use restriction endonucleases EcoRI and KpnI to perform double enzyme digestion, and recover the enzyme digestion product.

[0083] 3. Take the PBSK vector (Xincheng Xingchuang Technology Development Co., Ltd., product catalog number Biovector105802), use restriction endonucleases EcoRI and KpnI to perform double enzyme digestion, and recover the vector backbone.

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

[0085] 5. Take wild-type poplar leaves that have germinated for 3-4 weeks, cut the middle part of the leaves into 0.5-1 mm thin strips, put them into enzyme solution for dark digestion for 3-4 hours, centrifuge and collect protoplasts.

[0086] 6. Use the PEG method to transform the recombinant plasmid PBSK-PPG1 into protoplasts and culture them overnight under light.

[0087] 7. The expression of GFP in mesophyll cells was observed using a laser scanning confocal microscope (LSM510META, Zeiss). The objective magnification was 40 times, the excitation wavelength was 488 nm, the bandpass was BP505-530 nm, and the long pass was LP560 nm.

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

[0089] Example 3 Functional identification of PPG1 gene

[0090] 1. Construction of PPG1 gene overexpression vector

[0091] 1. Amplification of the target gene PPG1

[0092] pCAMBIA1300-35S-PPG1-F and pCAMBIA1300-35S-PPG1-R were synthesized respectively, and the PPG1 gene shown in SEQ ID NO.1 was amplified by PCR.

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

[0094] 5'-CCGggtaccATGGCAACTTTAACTCCTATTCAGACTT-3';

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

[0096] 5'-GGCggatccTTTATCAATTTGGTTGAGCTTGCCAAG-3'.

[0097] 2. The PPG1 fragment obtained in the previous step was double-digested with the pCAMBIA1300-35S vector using restriction endonucleases Kpn I and BamH I, and then ligated to obtain the pCAMBIA1300-35S-PPG1 recombinant vector. The pCAMBIA1300-35S-PPG1 recombinant vector was sequenced and verified.

[0098] The sequencing results showed that the pCAMBIA1300-35S-PPG1 recombinant vector was a pCAMBIA1300-35S vector in which the fragment between the Kpn I and BamH I restriction sites was replaced with the PPG1 gene, and the other sequences of the pCAMBIA1300-35S vector were kept unchanged. The obtained pCAMBIA1300-35S-ppg1 recombinant vector can overexpress the PPG1 protein.

[0099] 2. Obtaining PPG1 gene overexpression plants

[0100] 1. Take the PPG1 gene overexpression vector constructed in step 1 and introduce it into Agrobacterium GV3101 to obtain recombinant Agrobacterium.

[0101] 2. From plants that have been cultured for 1-3 months, select leaves with uniform leaf color and no abnormalities on the leaf surface. Remove the edges of the leaves and make even cuts in the middle of the leaves. Inoculate the recombinant Agrobacterium obtained in step 1 into the infection solution and adjust the concentration to OD600 = 0.3-0.4. Put the prepared leaves into the bacterial solution, shake continuously, infect for 15 minutes, take out the leaves, tear them into small pieces, inoculate them into the co-cultivation medium and culture them in the dark at 25°C for 2 days.

[0102] 3. Inoculate the co-cultivated leaves on the induction screening medium and continue to culture in the dark for about two weeks. After the callus grows, subculture to a new screening medium and subculture once every two weeks. Generally, the induction subculture is performed up to 3 times, and the longest cycle of the whole stage is 45 days. Inoculate the induced callus on the differentiation medium and subculture once every three weeks. During this period, the callus will turn green and harden. This stage normally takes about two months. Remove the seedlings that have grown to 1-3cm and inoculate them on the rooting screening medium. Roots can be formed in about 7-15 days. Transplant the normally grown plants into nutrient soil for culture, which is T 1 Generation of plants.

[0103] 4. From T 1 PPG1 gene overexpression plants were screened from the first generation plants.

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

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

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

[0107] 5. The PPG1 gene overexpressing plants obtained in step 4 are self-pollinated and seeds are harvested.

[0108] 6. Sow the seeds harvested in step 5 in a solid 1 / 2MS medium containing 50 mg / L hygromycin. After 10 days of cultivation, transplant the normally grown plants into nutrient soil for cultivation. 2 Generation of plants.

[0109] 7. From T 2 Screen the plants with overexpression of PPG1 gene from the generation plants (the specific steps are the same as step 4).

[0110] 8. The PPG1 gene overexpressing plants obtained in step 7 are self-pollinated and seeds are harvested.

[0111] 9. Sow the seeds harvested in step 8 in nutrient soil and cultivate them into plants, namely T 3 Generation of plants.

[0112] 10. T 3 The next generation of plants were identified (the specific steps are the same as step 4), and a certain T 2 The T 3 All the plants of the same generation were homozygous PPG1 overexpressing plants. 2 The plants of the first generation were named PPG1 overexpression plants, and the self-pollinated progeny of the PPG1 gene overexpression plants were named PPG1 gene overexpression lines.

[0113] 11. T of PPG1 gene overexpression strain 3 The first generation of plants are self-pollinated and seeds are harvested, and the harvested seeds are sown in nutrient soil and cultivated into plants, which are T 4 Generation of plants.

[0114] 12. T of PPG1 gene overexpression strain 4 The plants of the first generation are self-pollinated and seeds are harvested, and the harvested seeds are sown in nutrient soil and cultivated into plants, which are T 5 Generation of plants.

[0115] T of PPG1 gene overexpression plants and PPG1 gene overexpression lines 3 T of the generation plants and PPG1 gene overexpression lines 4 T of the generation plants and PPG1 gene overexpression lines s The plants of the next generation all had the following phenotypes: in the 1 / 2MS culture medium under suitable conditions, compared with the wild-type poplar, the plants of the PPG1 gene overexpression line grew better and had a significantly increased plant height.

[0116] 3. Poplar phenotypic and physiological identification

[0117] Poplars to be tested: T4 plants of the PPG1 gene overexpression strain (35S-PPG1-T 4 ) and wild-type (WT) poplar.

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

[0119] Under suitable conditions, the T of the PPG1 overexpressing strain germinated for 30 days 4 See the phenotype photos of the wild-type plants. Figure 3 .

[0120] Physiological parameters of the T4 plants of the PPG1 gene overexpression strain and wild-type plants were measured, including fresh weight and plant height. Figure 4 A and B in FIG. As shown in the figure, after being cultured for the same number of days under suitable conditions, the average fresh weight and plant height of the T4 generation plants of the four groups of PPG1 gene overexpression lines were significantly higher than those of the wild-type plants.

[0121] T of PPG1 overexpression lines 4 The photosynthetic parameters of the two generations of plants and wild-type plants were measured, including ETR, Fv / Fm, and NPQ. The results are shown in Figure 5 A, B and C in the figure. As shown in the figure, the T 4 The ETR, Fv / Fm, and NPQ of the first generation plants were higher than those of the wild-type plants. These data reflect that the PPG1 gene overexpression line has a stronger photosynthetic capacity than the wild-type plants.

[0122] These results showed that under suitable conditions, the T4 plants of the PPG1 overexpression line had higher plant height, stronger photosynthesis, larger biomass, and better growth than the wild-type plants in the same period of time. 4The growth rate and amount of the modified plants were higher than those of the wild-type plants. The above results showed that PPG1 protein and its coding sequence can promote the growth and development of poplar.

[0123] The embodiments described above are only preferred embodiments of the present invention and are only used to explain the present invention, not to limit the scope of implementation of the present invention. For those skilled in the art, other implementation methods can certainly be easily made by replacement or modification based on the technical contents disclosed in this specification. Therefore, all changes and improvements made on the principles of the present invention should be included in the scope of the patent application of the present invention.

[0124] Wherein, 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. A protein PPG1 for regulating the growth and development of poplar, characterized in that: The amino acid sequence of the protein PPG1 is shown in SEQ ID NO.

2.

2. The PPG1 gene encoding the protein PPG1 according to claim 1, characterized in that: The nucleotide sequence of the PPG1 gene is shown in SEQ ID NO.

1.

3. Use of the protein PPG1 according to claim 1 or the gene according to claim 2 in promoting the growth and development of poplar.

4. The use according to claim 3, characterized in that: The promotion of poplar growth and development is specifically manifested in that, compared with the wild type, the PPG1 gene overexpression strain has a higher plant height, stronger photosynthesis, larger biomass, and better growth.

5. Use of a gene expression cassette, a recombinant vector, a recombinant microorganism, and a transgenic plant cell line containing the PPG1 gene of claim 2 in promoting the growth and development of poplar.

6. A method for promoting the growth and development of poplars, characterized in that: The method comprises: (1) By increasing the activity of protein PPG1 in poplar or increasing the content of protein PPG1 in poplar; (2) by promoting the expression of the PPG1 gene in poplar; The amino acid sequence of the protein PPG1 is shown in SEQ ID NO.2, and the nucleotide sequence of the PPG1 gene is shown in SEQ ID NO.

1.

7. The method for promoting the growth and development of poplars according to claim 6, characterized in that: The way to promote the expression of PPG1 gene in poplar is overexpression or overexpression.

8. The method for promoting the growth and development of poplars according to claim 7, characterized in that: The PPG1 gene in the plant genome is overexpressed to obtain PPG1 gene overexpressing plants, and homozygous PPG1 gene overexpressing plants are obtained from the self-pollinated offspring of the PPG1 gene overexpressing plants; the homozygous PPG1 gene overexpressing plants are plants that grow and develop faster.

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