Application of OsPL3 protein in regulation and control of salt tolerance and root length of rice

Abstract

The invention discloses application of OsPL3 protein in regulation and control of salt tolerance and root length of rice. The invention belongs to the technical field of biology, and particularly relates to application of OsPL3 protein to regulation and control of salt tolerance and root length of rice. The protein provided by the invention can positively regulate and control the salt tolerance and root length of rice, through the expression quantity of the overexpressed OsPL3 protein, after salt stress treatment, the survival rate of seedlings of the obtained overexpressed plant OsPL3-OE is obviously increased by about one time compared with that of a wild type, the root length development of the OsPL3-OE is also obviously longer than that of the wild type, and the protein has a certain application prospect for rice breeding.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to application of OsPL3 protein in regulating rice salt tolerance and root length. Background Art

[0002] In recent years, the rapid development of biotechnology has greatly promoted the innovation of plant breeding research methods and the continuous improvement of research levels. Plant disease and insect resistance and herbicide resistance biotechnology breeding has begun to enter the practical stage. The use of biotechnology to introduce exogenous insecticide and herbicide resistance genes into the plant genome has broken the natural barrier that makes it difficult for plant species and even species to hybridize, and achieved the transfer of insect resistance and herbicide resistance genes, so that plants can quickly and directionally acquire insect resistance and mechanized weed control, while retaining the original good agronomic traits. Summary of the invention

[0003] The technical problem solved by the invention is how to regulate the salt tolerance and root length of plants, especially rice.

[0004] In order to solve the above problems, the present invention provides related applications of proteins, substances for regulating the expression of genes encoding the proteins, or substances for regulating the activity or content of the proteins.

[0005] Use of the protein provided by the present invention, a substance for regulating the expression of a gene encoding the protein, or a substance for regulating the activity or content of the protein in any of the following:

[0006] 1) Application in regulating plant salt tolerance and root length;

[0007] 2) Application in the preparation of products for regulating plant salt tolerance and root length;

[0008] 3) Application in breeding plants with salt tolerance and altered root length;

[0009] 4) Application in the preparation of products for cultivating plants with altered salt tolerance and root length;

[0010] 5) Application in plant breeding.

[0011] The protein is any of the following:

[0012] a1) a protein having an amino acid sequence of SEQ ID No. 1;

[0013] a2) a protein having the same function as the amino acid sequence shown in SEQ ID No. 1 after one or more amino acid residues are replaced and / or deleted and / or added;

[0014] a3) a protein having an amino acid sequence with more than 80% identity with any one of a1) to (a2) and having the same function;

[0015] a4) A fusion protein obtained by connecting a tag to the end of the protein defined in any one of a1) to (a3).

[0016] In the above proteins, the protein tag refers to a polypeptide or protein that is fused and expressed with the target protein using DNA in vitro recombination technology to facilitate the expression, detection, tracing and / or purification of the target protein. The protein tag can be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag and / or a SUMO tag, etc.

[0017] In the above-mentioned proteins, identity refers to the identity of the amino acid sequence. The identity of the amino acid sequence can be determined using a homology search site on the Internet, such as the BLAST page on the NCBI homepage website. For example, in Advanced BLAST2.1, by using blastp as a program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as a Matrix, setting the Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default values) respectively, and searching for the identity of a pair of amino acid sequences, the identity value (%) can be obtained.

[0018] In the above proteins, the above 80% identity may be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

[0019] Among the above proteins, SEQ ID No. 1 consists of 895 amino acid residues and is named as OsPL3 protein, and its encoding gene is OsPL3 gene.

[0020] In the above application, the protein is derived from rice (Oryza sativa L.).

[0021] Herein, the substance that regulates the activity and / or content of the protein may be a substance that regulates the expression of a gene encoding the protein OsPL3.

[0022] In the above, the substance that regulates gene expression may be a substance that performs at least one of the following six types of regulation: 1) regulation at the transcription level of the gene; 2) regulation after transcription of the gene (that is, regulation of the splicing or processing of the primary transcript of the gene); 3) regulation of RNA transport of the gene (that is, regulation of the transport of the mRNA of the gene from the nucleus to the cytoplasm); 4) regulation of the translation of the gene; 5) regulation of the degradation of the mRNA of the gene; 6) post-translational regulation of the gene (that is, regulation of the activity of the protein translated from the gene).

[0023] In the present invention, the regulation may be up-regulation, enhancement or increase; the regulation may also be down-regulation, attenuation or reduction.

[0024] Herein, the enhancement, increase or upregulation of the expression level of the gene encoding the protein mentioned above in the recipient plant, or / and the enhancement, increase or upregulation of the activity and / or content of the gene encoding the above protein is achieved by introducing the gene encoding the above protein into the recipient plant.

[0025] Herein, the regulation of the expression of the coding gene of the protein may be inhibition, reduction or down-regulation of the coding gene expression. Inhibition, reduction or down-regulation of the coding gene expression may be achieved by gene knockout or gene silencing.

[0026] In the above application, the substance that regulates the expression of the gene encoding the protein or the substance that regulates the activity or content of the protein may be a biological material related to the protein mentioned above, and the biological material may be any of the following:

[0027] c1) a nucleic acid molecule encoding the protein described above;

[0028] c2) an expression cassette containing the nucleic acid molecule described in c1);

[0029] c3) a recombinant vector containing the nucleic acid molecule described in c1), or a recombinant vector containing the expression cassette described in c2);

[0030] c4) a recombinant microorganism containing the nucleic acid molecule described in c1), or a recombinant microorganism containing the expression cassette described in c2), or a recombinant microorganism containing the recombinant vector described in c3);

[0031] c5) a transgenic plant cell line containing the nucleic acid molecule described in c1), or a transgenic plant cell line containing the expression cassette described in c2);

[0032] c6) transgenic plant tissue containing the nucleic acid molecule described in c1), or transgenic plant tissue containing the expression cassette described in c2);

[0033] c7) a transgenic plant organ containing the nucleic acid molecule described in c1), or a transgenic plant organ containing the expression cassette described in c2);

[0034] e1) a nucleic acid molecule that inhibits, reduces or silences the expression of the protein encoding gene mentioned above;

[0035] e2) an expression cassette containing the nucleic acid molecule described in e1);

[0036] e3) a recombinant vector containing the nucleic acid molecule described in e1), or a recombinant vector containing the expression cassette described in e2);

[0037] e4) a recombinant microorganism containing the nucleic acid molecule described in e1), or a recombinant microorganism containing the expression cassette described in e2), or a recombinant microorganism containing the recombinant vector described in e3);

[0038] e5) a transgenic plant cell line containing the nucleic acid molecule described in e1), or a transgenic plant cell line containing the expression cassette described in e2);

[0039] e6) transgenic plant tissue containing the nucleic acid molecule described in e1), or transgenic plant tissue containing the expression cassette described in e2);

[0040] e7) A transgenic plant organ containing the nucleic acid molecule described in e1), or a transgenic plant organ containing the expression cassette described in e2).

[0041] In the above application, the nucleic acid molecule described in c1) can be any of the following DNA molecules,

[0042] d1) the nucleotide sequence is the DNA molecule shown in SEQ ID No.3;

[0043] d2) the coding sequence is the DNA molecule shown in SEQ ID No.2;

[0044] d3) a DNA molecule that has 90% or more identity with the nucleotide sequence defined in d1) or d2) and encodes the protein described above;

[0045] d4) A DNA molecule that hybridizes with the nucleotide sequence defined in d1) or d2) under stringent conditions and encodes the protein described above.

[0046] In the above application, the nucleic acid molecule described in e1) may be a DNA molecule whose nucleotide sequence is shown as SEQ ID No.3.

[0047] The nucleic acid molecule described herein can be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule can also be RNA, such as gRNA, mRNA, siRNA, shRNA, sgRNA, miRNA or antisense RNA.

[0048] The vectors described herein are well known to those skilled in the art, including but not limited to: plasmids, phages (such as lambda phage or M13 filamentous phage, etc.), cosmids (i.e., cosmids), Ti plasmids or viral vectors. Specifically, the vector may be pSuper1300.

[0049] The recombinant expression vector containing the OsPL3 gene can be constructed using existing plant expression vectors. The plant expression vectors include, but are not limited to, binary Agrobacterium vectors and vectors that can be used for plant microprojectile bombardment. The plant expression vectors may also contain the 3' non-translated region of the foreign gene, i.e., a polyadenylic acid signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylic acid signal can guide the addition of polyadenylic acid to the 3' end of the mRNA precursor, such as, but not limited to, Agrobacterium crown gall induction (Ti) plasmid genes (such as rouge synthase Nos gene), plant genes (such as soybean storage protein gene) 3' end transcription non-translated regions have similar functions.

[0050] When using the OsPL3 gene to construct a recombinant plant expression vector, any enhanced promoter or constitutive promoter can be added before its transcription initiation nucleotide, including but not limited to cauliflower mosaic virus (CAMV) 35S promoter, corn ubiquitin promoter (ubiquitin), which can be used alone or in combination with other plant promoters; in addition, when using the gene of the present invention to construct a plant expression vector, enhancers can also be used, including translation enhancers or transcription enhancers, and these enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signal and the start codon are extensive, and can be natural or synthetic. The translation initiation region can come from the transcription initiation region or the structural gene.

[0051] In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used can be processed, such as adding genes that can be expressed in plants and encode enzymes or luminescent compounds that can produce color changes (GUS gene, luciferase gene, etc.), antibiotic markers with resistance (gentamicin marker, kanamycin marker, etc.) or chemical agent resistance marker genes (such as herbicide resistance genes), etc. Considering the safety of transgenic plants, no selective marker genes can be added, and transformed plants can be directly screened by adversity.

[0052] The present invention also provides a method for changing the salt tolerance and root length of a plant, the method comprising the following steps M or P:

[0053] The step M is to enhance, increase or upregulate the activity and / or content of the protein mentioned above in the target plant, or / and enhance, increase or upregulate the expression of the gene encoding the protein mentioned above, so as to improve the salt tolerance and root length of the plant;

[0054] The method comprises step P, wherein step P is to inhibit, reduce or silence the activity and / or content of the protein mentioned above in the target plant, or / and, to inhibit, reduce or silence the expression level of the gene encoding the protein mentioned above, so as to reduce the salt tolerance and root length of the plant.

[0055] In the above method, reducing the expression level and / or activity of the gene encoding the protein OsPL3 in the target plant can be achieved by using gene mutation, gene knockout, gene editing or gene knockdown technology to reduce or inactivate the activity of the gene encoding the protein OsPL3 in the genome of the target plant.

[0056] The present invention also provides a method for cultivating plants with improved salt tolerance and elongated roots, comprising up-regulating or enhancing or increasing the expression level of the gene encoding the protein mentioned above in the target plant, and / or the activity and / or content of the protein to obtain plants with improved salt tolerance and elongated roots, wherein the plants with improved salt tolerance and elongated roots have higher salt tolerance than the target plant and have longer roots than the target plant.

[0057] In a specific embodiment, the upregulation, enhancement or increase of the expression of the gene encoding the protein described above in the plant comprises introducing the nucleic acid molecule, expression cassette or recombinant vector described above into the target plant to obtain a plant with high salt tolerance and root length.

[0058] The present invention also provides a method for cultivating plants with reduced salt tolerance and shortened roots, comprising inhibiting or reducing or silencing the expression level of the gene encoding the protein mentioned above in the target plant, and / or the activity and / or content of the protein to obtain plants with reduced salt tolerance and shortened roots, wherein the plants with reduced salt tolerance have lower salt tolerance than the target plant, and the plants with shortened roots have shorter roots than the target plant.

[0059] In a specific embodiment, the inhibition, reduction or silencing of the expression of the gene encoding the protein described above in the plant comprises introducing into the target plant the nucleic acid molecule, expression cassette or recombinant vector that inhibits, reduces or silences the protein described above, thereby obtaining a plant with reduced salt tolerance and shortened roots.

[0060] Herein, the breeding purpose includes cultivating plants with high salt tolerance and elongated roots; the breeding purpose also includes cultivating plants with low salt tolerance and shortened root length.

[0061] The high salt tolerance and root length plants have improved salt tolerance and extended root length compared to the target plants. The low salt tolerance and root length plants have reduced salt tolerance and shortened root length compared to the target plants.

[0062] Herein, the improvement of salt tolerance can be an improvement of survival rate. The survival rate can be a survival rate of seedlings in the seedling stage.

[0063] Herein, the rice may be rice ZH11.

[0064] Herein, the recombinant microorganism may be Agrobacterium EHA105.

[0065] The proteins and / or biological materials mentioned above also fall within the scope of protection claimed by the present invention.

[0066] In the above application or method, the plant may be any of the following:

[0067] N1) Monocotyledonous plants;

[0068] N2) Gramineae;

[0069] N3) Gramineae;

[0070] N4) Oryza plants;

[0071] N5) Rice.

[0072] The present invention discloses the regulatory effect of OsPL3-OE protein and its encoding gene on plant stress tolerance, growth performance and production performance. The present invention protects the application of OsPL3 protein in regulating plant stress tolerance, and the regulation is positive regulation, that is, the content of OsPL3 protein increases and the salt tolerance of plants is enhanced. The present invention also protects the application of OsPL3 protein in regulating plant growth performance and / or production performance. The present invention can be used for improving plant stress tolerance, and has great application and promotion value for plant breeding, especially rice breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] Figure 1 Schematic diagram of the vector structure for constructing an overexpression strain.

[0074] Figure 2 is the OsPL3 gene expression level of the overexpression strain.

[0075] Figure 3 Rice OsPL3 positively regulates rice root growth and development. A shows the seedling phenotypes of the wild type and OsPL3-OE after one week of growth; B shows the root length statistics of the wild type and OsPL3-OE.

[0076] Figure 4The phenotypes and survival rate statistics of wild type and OsPL3 overexpression strains before and after salt stress treatment. A is the phenotype diagram of wild type and OsPL3 overexpression strains before and after salt stress treatment; B is the statistical diagram of the survival rate of wild type and OsPL3 overexpression strains after salt stress treatment. DETAILED DESCRIPTION

[0077] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0078] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0079] Unless otherwise specified, the quantitative tests in the following examples were performed three times and the results were averaged.

[0080] pSuper1300 in the following examples has been described in: Li, Cong et al. "Mutual upregulation of HY5 and TZP in mediating phytochrome A signaling." The Plant cell vol. 34, 1 (2022): 633-654. doi: 10.1093 / plcell / koab254. The public can obtain the biological material from the applicant, and the biological material is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0081] Agrobacterium EHA105 in the following examples has been described in: C. Zhang, X. Du, K. Tang, Z. Yang, L. Pan, P. Zhu, J. Luo, Y. Jiang, H. Zhang, H. Wan, X. Wang, F. Wu, WA Tao, X.-J. He, H. Zhang, RA Bressan, J. Du, JK Zhu, Arabidopsis A GDP1 links H3K9me2 to DNA methylation inheterochromatin. Nat. Commun. 9, 4547 (2018). doi: 10.1038 / s41467-018-06965-w Medlin. The public can obtain the biological material from the applicant, which is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0082] The rice ZH11 in the following example has been recorded in: Qin, Hua et al. "Orchestration of ethylene and gibberellin signals determines primary root elongation in rice." The Plant cell vol. 34, 4 (2022): 1273-1288. doi: 10.1093 / plcell / koac008. The public can obtain the biological material from the applicant, and the biological material is only used for repeating the experiment of the present invention and cannot be used for other purposes.

[0083] The following examples were processed using SPSS11.5 statistical software, and the experimental results were expressed as mean ± standard deviation. One-way ANOVA test was used, and P < 0.05 (*) indicated a significant difference, P < 0.01 (**) indicated a very significant difference, and P < 0.001 (***) indicated an extremely significant difference.

[0084] Example 1. Obtaining the Rice OsPL3 Gene

[0085] RNA was extracted from leaves of rice variety (Nip) and reverse transcribed into cDNA. Using the cDNA as a template, PCR amplification was performed using primers OsPL3-F: 5'-ATGAGATCTGTCTCTGAGCT-3'; OsPL3-R: 5'-AAAACGAAGCAGCTCTCTTT-3' and MaxSuper-Fidelity DNA Polymerase (Cat. No.: P505-d1, Vazyme) to obtain the amplified product (i.e., the coding region of the OsPL3 gene).

[0086] The coding sequence (CDS) of the OsPL3 gene in the rice variety Nip is SEQ ID No. 2, and the encoded amino acid sequence is OsPL3 shown in SEQ ID No. 1. In the genomic DNA of rice Nip, the gene encoding the OsPL3 protein is shown in SEQ ID No. 3 in the sequence table. The 1230-1551 positions of SEQ ID No. 3 are the first exon, the 1929-2063 positions are the second exon, the 2131-4045 positions are the third exon, the 4692-4807 positions are the fourth exon, the 4924-5015 positions are the fifth exon, and the 5168-5275 positions are the sixth exon.

[0087] Example 2: Construction of recombinant plasmid pSuper1300-OsPL3-GFP

[0088] Using Nip cDNA as template, PCR was performed to amplify the coding region sequence of OsPL3 with primers OsPL3-F / OsPL3-R, and the target gene fragment of OsPL3 was recovered by gel.

[0089] The target gene and vector were digested with the same restriction endonuclease (XbaI), ligated and transformed into E. coli, and then the pSuper1300-OsPL3-GFP overexpression vector was obtained by first-generation sequencing verification and sequence alignment. The schematic diagram of the structure of the recombinant plasmid pSuper1300-OsPL3-GFP backbone is shown in Figure 1 .

[0090] The recombinant vector pSuper1300-OsPL3-GFP is obtained by replacing a small fragment of the restriction enzyme XbaI enzyme recognition site of the pSuper1300 vector with a DNA molecule (OsPL3 encoding gene) having a nucleotide sequence of SEQ ID No.2, and keeping other sequences of the pSuper1300 vector unchanged. After full plasmid sequencing, the recombinant plasmid pSuper1300-OsPL3-GFP is shown as SEQ ID No.4.

[0091] Example 3: Obtaining transgenic rice

[0092] The recombinant plasmid pSuper1300-OsPL3-GFP obtained in Example 2 was introduced into Agrobacterium tumefaciens EHA105 to obtain recombinant Agrobacterium. The recombinant Agrobacterium was used to genetically transform the embryonic callus of rice ZH11 by the Agrobacterium infection method, and then the resistant callus was screened (100 mg / L hygromycin was used for resistance screening), and then differentiation regeneration culture was carried out, and then rooting culture was carried out to obtain regenerated plants.

[0093] The specific steps are as follows:

[0094] (1) Take out the mature seeds of the plant, remove the shells, and select the plump, smooth and sterile seeds for disinfection.

[0095] (2) Inoculate the sterilized seeds onto the induction medium and culture them in the dark at 28°C for about 14 days. Select the callus with good appearance and good growth ability.

[0096] (3) The recombinant vector pSuper1300-OsPL3-GFP constructed in Example 2 was introduced into Agrobacterium tumefaciens EHA105 to obtain a recombinant bacterium named EHA105 / pSuper1300-OsPL3-GFP.

[0097] (4) Take the recombinant bacteria obtained in step (3), resuspend the bacteria in infection medium, and obtain EHA105 / pSuper1300-OsPL3-GFP bacterial suspension.

[0098] (5) Soak the Nip callus tissue from step (2) in the EHA105 / pSuper1300-OsPL3-GFP bacterial suspension prepared in step (4) for infection for 20 minutes. After infection, pour out the bacterial suspension, take the callus tissue, dry it with sterile filter paper, and then place it on a co-culture medium with acetosyringone and glucose, and culture it in the dark at 28°C for 50-55 hours.

[0099] (6) After completing step (5), callus tissues without obvious Agrobacterium on the surface were selected and transferred to an antibacterial medium containing cephalosporin, and cultured in the dark at 28° C. for 3-4 days.

[0100] (7) The cultured callus tissue was transferred to a screening medium containing hygromycin and cephalosporin (the concentration of hygromycin was 100 mg / L and the concentration of cephalosporin was 100 mg / L) and cultured in the dark at 28°C for 30 days, with subculturing every 10 days.

[0101] (8) After completing step (7), fresh hygromycin-resistant callus tissue was taken and inoculated into a pre-regeneration medium, cultured in the dark at 28°C for 7 days, then placed in a light culture room (12 h light / 12 h dark) for another 7 days, and then transferred to a regeneration medium and continued to be cultured in the light until regenerated plants grew to obtain OsPL3-OE transgenic plants. The culture medium and formulation used for genetic transformation: The induction medium and differentiation medium formulations were both MS medium.

[0102] The transgenic plants obtained using the recombinant vector pSuper1300-OsPL3-GFP were recorded as OsPL3 transgenic plants.

[0103] Example 4: Identification of transgenic rice and detection of gene expression levels

[0104] 1. Identification of OsPL3 transgenic rice

[0105] Plants to be tested: rice Nip (CK) and OsPL3 transgenic plants obtained in Example 3.

[0106] The genomic DNA of the plant to be tested was extracted, and the genomic DNA was used as a template, and the primer pair consisting of primers OsPL3-F / OsPL3-R was used for PCR amplification, and the pSuper1300-OsPL3-GFP plasmid was used as a positive control (V), and the recipient variety Nip was used as a negative control (CK). The obtained product was then sequenced.

[0107] 2. Detection of expression level of OsPL3 transgenic rice

[0108] The test plants were cultured under parallel conditions, specifically: the test rice seeds were taken, germinated and seedlings were raised in a greenhouse (starting from the time of whitening, and cultured for 3 weeks), the seedling materials were quickly frozen in liquid nitrogen, RNA was extracted (TIANGEN kit number: DP419), reverse transcribed (NoviZan kit: number: R333-01), and fluorescent quantitative PCR was used to detect gene expression. The detection primers are as follows:

[0109] OsPL3-RT-F: 5'-GTCCGTGTCTGCTGTGAATC-3';

[0110] OsPL3-RT-R: 5'-GCTTTTGTACTTGGGCAGCT-3'.

[0111] Depend on Figure 2 It can be seen that the gene expression levels of different OsPL3 transgenic rice were significantly increased.

[0112] Example 5: Study on plant root phenotype and salt tolerance

[0113] The tested seeds were: rice Nipponbare seeds (Nip) and transgenic plants OsPL3-OE1 (abbreviated as PL3-OE1), OsPL3-OE2 (abbreviated as PL3-OE2), and OsPL3-OE3 (abbreviated as PL3-OE3).

[0114] 1. Take the test rice seeds, germinate and grow seedlings in the greenhouse (starting from the time of whitening, and culture for a total of 1 week), obtain 1-week seedlings, take photos; and count the root length.

[0115] Depend on Figure 3 As shown in A and B, the root length of OsPL3-OE is significantly longer than that of the wild type.

[0116] 2. Cultivate the test plants under parallel conditions, specifically: take the test rice seeds, germinate and grow seedlings in the greenhouse (starting from the time of white appearance, a total of 3 weeks of cultivation), obtain 3-week seedlings, and take pictures; treat the 3-week seedlings with 150mM NaCl for 5 days and take pictures for statistical phenotype and survival rate statistics. Figure 4 The greenhouse conditions were: 28°C, 10 h light / 14 h dark.

[0117] Depend on Figure 4 As shown in Figures A and B, after salt stress treatment, the survival rate of OsPL3-OE seedlings was significantly increased by about 1 times compared with the wild type, indicating that OsPL3 positively regulates the salt stress response of rice.

[0118] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.

Claims

1. Use of a protein or a substance that regulates the expression of a gene encoding the protein or a substance that regulates the activity or content of the protein in any of the following; 1) Application in regulating plant salt tolerance and root length; 2) Application in the preparation of products for regulating plant salt tolerance and root length; 3) Application in breeding plants with salt tolerance and altered root length; 4) Application in the preparation of products for cultivating plants with altered salt tolerance and root length; 5) Application in plant breeding; The protein is any of the following: a1) a protein having an amino acid sequence of SEQ ID No. 1; a2) a protein having the same function as the amino acid sequence shown in SEQ ID No. 1 after one or more amino acid residues are replaced and / or deleted and / or added; a3) a protein having an amino acid sequence identity of more than 80% with that of a1) or a2) and having the same function; a4) A fusion protein obtained by connecting a tag to the end of the protein defined in any one of a1) to a3).

2. The use according to claim 1, characterized in that: The protein is derived from rice.

3. The use according to claim 1 or 2, characterized in that: The substance that regulates gene expression or the substance that regulates the activity or content of the protein is a biological material related to the protein in the application according to claim 1 or 2, and the biological material is any one of the following: c1) a nucleic acid molecule encoding the protein; c2) an expression cassette containing the nucleic acid molecule described in c1); c3) a recombinant vector containing the nucleic acid molecule described in c1), or a recombinant vector containing the expression cassette described in c2); c4) a recombinant microorganism containing the nucleic acid molecule described in c1), or a recombinant microorganism containing the expression cassette described in c2), or a recombinant microorganism containing the recombinant vector described in c3); c5) a transgenic plant cell line containing the nucleic acid molecule described in c1), or a transgenic plant cell line containing the expression cassette described in c2); c6) transgenic plant tissue containing the nucleic acid molecule described in c1), or transgenic plant tissue containing the expression cassette described in c2); c7) a transgenic plant organ containing the nucleic acid molecule described in c1), or a transgenic plant organ containing the expression cassette described in c2); e1) a nucleic acid molecule that inhibits, reduces or silences the expression of the protein encoding gene; e2) an expression cassette containing the nucleic acid molecule described in e1); e3) a recombinant vector containing the nucleic acid molecule described in e1), or a recombinant vector containing the expression cassette described in e2); e4) a recombinant microorganism containing the nucleic acid molecule described in e1), or a recombinant microorganism containing the expression cassette described in e2), or a recombinant microorganism containing the recombinant vector described in e3); e5) a transgenic plant cell line containing the nucleic acid molecule described in e1), or a transgenic plant cell line containing the expression cassette described in e2); e6) transgenic plant tissue containing the nucleic acid molecule described in e1), or transgenic plant tissue containing the expression cassette described in e2); e7) A transgenic plant organ containing the nucleic acid molecule described in e1), or a transgenic plant organ containing the expression cassette described in e2).

4. The use according to claim 3, characterized in that: c1) The nucleic acid molecule is a DNA molecule as shown in any of the following: d1) the nucleotide sequence is the DNA molecule shown in SEQ ID No.3; d2) the coding sequence is the DNA molecule shown in SEQ ID No.2; d3) a DNA molecule having 90% or more identity with the nucleotide sequence defined in d1) or d2) and encoding the protein described in claim 1; d4) A DNA molecule which hybridizes with the nucleotide sequence defined in d1) or d2) under stringent conditions and encodes the protein described in claim 1.

5. A method for changing the salt tolerance and root length of plants, characterized in that: The method comprises steps M or P: The step P is to enhance, increase or up-regulate the activity and / or content of the protein described in claim 1 or 2 in the target plant, or / and enhance, increase or up-regulate the expression of the gene encoding the protein described in claim 1 or 2 to improve the salt tolerance and root length of the plant; The step M is to inhibit, reduce or silence the activity and / or content of the protein described in claim 1 or 2 in the target plant, or / and, inhibit, reduce or down-regulate the expression level of the gene encoding the protein described in claim 1 or 2, so as to reduce the plant's salt tolerance and root length.

6. A method for cultivating plants with improved salt tolerance and elongated roots, characterized in that: The invention comprises enhancing, increasing or up-regulating the expression level of the gene encoding the protein described in claim 1 or 2 in the target plant, and / or the activity and / or content of the protein to obtain a plant with improved salt tolerance and elongated roots, wherein the plant with improved salt tolerance and elongated roots has a higher salt tolerance than the target plant and has longer roots than the target plant.

7. The method according to claim 6, characterized in that The enhancing, increasing or up-regulating the expression of the gene encoding the protein described in claim 1 or 2 in the plant comprises introducing the nucleic acid molecule described in c1) of claim 4, the expression cassette described in c2) or the recombinant vector described in c3) into the target plant to obtain a plant with improved salt tolerance and root length.

8. A method for cultivating plants with reduced salt tolerance and shortened roots, characterized in that: The method comprises inhibiting, reducing or silencing the expression level of the gene encoding the protein described in claim 1 or 2 in the target plant, and / or reducing the activity and / or content of the protein to obtain a plant with reduced salt tolerance and shortened roots, wherein the plant with reduced salt tolerance has a lower salt tolerance than the target plant, and the plant with shortened roots has shorter roots than the target plant.

9. The protein according to claim 1 or 2 and / or the biomaterial according to claim 3 or 4.

10. The method according to any one of claims 5 to 8, characterized in that: The plant is any of the following: N1) Monocots; N2) Gramineae; N3) Gramineae; N4) Oryza plants; N5) Rice.

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