Application of ospl3 protein in regulating salt tolerance and root length of rice
By introducing and regulating the expression of the gene encoding the OsPL3 protein, and using recombinant vectors and gene editing technology, the problems of salt tolerance and root length regulation in rice were solved, resulting in enhanced salt tolerance and improved root growth performance.
Patent Information
- Application Number
- CN202510074542.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-17
- Publication Date
- 2025-12-16
- Estimated Expiration
- 2045-01-17
AI Technical Summary
How to regulate the salt tolerance and root length of plants, especially rice.
By introducing and regulating the expression or activity of the gene encoding the OsPL3 protein, recombinant vectors and gene editing technologies can be used to enhance or inhibit the expression and activity of the OsPL3 protein, thereby altering the salt tolerance and root length of plants.
This study improved the salt tolerance of rice and regulated root length, thereby enhancing the salt tolerance and root growth performance of rice and increasing the survival rate of seedlings.
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Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biotechnology, and particularly relates to application of an OsPL3 protein in regulating salt tolerance and root length of rice. BACKGROUND
[0002] In recent years, the rapid development of biotechnology has greatly promoted the innovation of plant breeding research means and the continuous improvement of research level. Plant disease and pest resistance and herbicide resistance biotechnology breeding has begun to enter the practical stage. By using biotechnology means, an exogenous insecticidal and herbicide-resistant gene is introduced into a plant genome, the natural barrier of plant species or even species is broken, and the transfer of the insecticidal and herbicide-resistant gene is realized, so that the plant rapidly and directionally obtains insect resistance and mechanical herbicide, and meanwhile, the original good agronomic traits are retained. SUMMARY
[0003] The technical problem solved by the application is how to regulate the salt tolerance and root length of plants, especially rice.
[0004] In order to solve the above problems, the application provides related applications of a protein, a substance for regulating expression of a coding gene of the protein, or a substance for regulating activity or content of the protein.
[0005] The application provides applications of the protein, the substance for regulating expression of the coding gene of the protein, or the substance for regulating activity or content of the protein in any one of the following:
[0006] 1) in regulating salt tolerance and root length of plants;
[0007] 2) in preparing a product for regulating salt tolerance and root length of plants;
[0008] 3) in cultivating plants with changed salt tolerance and root length;
[0009] 4) in preparing a product for cultivating plants with changed salt tolerance and root length;
[0010] 5) in plant breeding.
[0011] The protein is any one of the following proteins:
[0012] a1) a protein with an amino acid sequence of SEQ ID No. 1;
[0013] a2) a protein with an amino acid sequence of SEQ ID No. 1, which has one or more substitutions, deletions and / or additions of amino acid residues and has the same function;
[0014] a3) a protein having 80% or more identity to any one of the amino acid sequences defined in (a1)-(a2) and having the same function;
[0015] a4) a fusion protein obtained by linking a terminal tag to any one of the proteins defined in (a1)-(a3).
[0016] In the above protein, the protein tag refers to a polypeptide or protein fused and expressed with a target protein by using DNA in vitro recombination technology, so as 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 protein, the identity refers to the identity of the amino acid sequence. The identity of the amino acid sequence can be determined by using the homology search site on the Internet, such as the BLAST webpage of the NCBI homepage. For example, the identity of a pair of amino acid sequences can be calculated by using blastp as the program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as the Matrix, setting Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default value) respectively, and performing a search in Advanced BLAST 2.1, and then the value of the identity (%) can be obtained.
[0018] In the above protein, the 80% or more identity can be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.
[0019] In the above protein, SEQ ID No. 1 consists of 895 amino acid residues. It 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 can be a substance that regulates the expression of a gene encoding the protein OsPL3.
[0022] In the above, the substance that regulates the expression of the gene can be a substance that performs at least one of the following six regulations: 1) regulation at the transcription level of the gene; 2) regulation after the transcription of the gene (i.e. regulation of the splicing or processing of the primary transcript of the gene); 3) regulation of the RNA transport of the gene (i.e. 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 mRNA degradation of the gene; and 6) post-translational regulation of the gene (i.e. regulation of the activity of the protein translated from the gene).
[0023] In the present application, the regulation can be up-regulation or enhancement or increase; the regulation can also be down-regulation or weakening or decrease.
[0024] In the present application, the enhancement, increase or up-regulation of the expression of the gene encoding the protein as described above in the recipient plant, or / and the enhancement, increase or up-regulation of the activity and / or content of the gene encoding the protein as described above is achieved by introducing the gene encoding the protein as described above into the recipient plant.
[0025] In the present application, the regulation of the expression of the gene encoding the protein as described above can be inhibition or decrease or down-regulation of the expression of the gene. The inhibition or decrease or down-regulation of the expression of the gene can 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 as described above or the substance that regulates the activity or content of the protein as described above can be a biological material related to the protein as described above, and the biological material can be any one of the following:
[0027] c1) a nucleic acid molecule encoding the protein as described above;
[0028] c2) an expression cassette containing the nucleic acid molecule of c1);
[0029] c3) a recombinant vector containing the nucleic acid molecule of c1), or a recombinant vector containing the expression cassette of c2);
[0030] c4) a recombinant microorganism containing the nucleic acid molecule of c1), or a recombinant microorganism containing the expression cassette of c2), or a recombinant microorganism containing the recombinant vector of c3);
[0031] c5) a transgenic plant cell line containing the nucleic acid molecule of c1), or a transgenic plant cell line containing the expression cassette of c2);
[0032] c6) a transgenic plant tissue containing the nucleic acid molecule of c1), or a transgenic plant tissue containing the expression cassette of c2);
[0033] c7) a transgenic plant organ comprising the nucleic acid molecule of c1), or a transgenic plant organ comprising the expression cassette of c2);
[0034] e1) a nucleic acid molecule which inhibits or reduces or silences the expression of a gene encoding the protein as described above;
[0035] e2) an expression cassette comprising the nucleic acid molecule of e1);
[0036] e3) a recombinant vector comprising the nucleic acid molecule of e1), or a recombinant vector comprising the expression cassette of e2);
[0037] e4) a recombinant microorganism comprising the nucleic acid molecule of e1), or a recombinant microorganism comprising the expression cassette of e2), or a recombinant microorganism comprising the recombinant vector of e3);
[0038] e5) a transgenic plant cell line comprising the nucleic acid molecule of e1), or a transgenic plant cell line comprising the expression cassette of e2);
[0039] e6) a transgenic plant tissue comprising the nucleic acid molecule of e1), or a transgenic plant tissue comprising the expression cassette of e2);
[0040] e7) a transgenic plant organ comprising the nucleic acid molecule of e1), or a transgenic plant organ comprising the expression cassette of e2).
[0041] In the above uses, the nucleic acid molecule of c1) can be any one of the following DNA molecules,
[0042] d1) a DNA molecule whose nucleotide sequence is set forth in SEQ ID No. 3;
[0043] d2) a DNA molecule whose coding sequence is set forth in SEQ ID No. 2;
[0044] d3) a DNA molecule which has 90% or more identity with the nucleotide sequence defined in d1) or d2), and which encodes the protein as described above;
[0045] d4) a DNA molecule which hybridizes to the nucleotide sequence defined in d1) or d2) under stringent conditions, and which encodes the protein as described above.
[0046] In the above uses, the nucleic acid molecule of e1) can be a DNA molecule whose nucleotide sequence is set forth in 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 and include, but are not limited to, plasmids, bacteriophages (e.g., lambda phage or M13 filamentous phage, etc.), cosmids (i.e., cosmids), Ti plasmids, or viral vectors. Specifically, the vector can 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, etc. The plant expression vector can also contain a 3' untranslated region of the foreign gene, i.e., a polyadenylation signal and any other DNA segment involved in mRNA processing or gene expression. The polyadenylation signal can direct the addition of polyadenylic acid to the 3' end of the mRNA precursor, such as the 3' untranslated region of the Agrobacterium tumefaciens (Ti) plasmid gene (e.g., the nopaline synthase (Nos) gene), the plant gene (e.g., the soybean storage protein gene), etc.
[0050] When constructing the recombinant plant expression vector using the OsPL3 gene, any one of the enhancer promoters or constitutive promoters can be added before the transcription initiation nucleotide, including but not limited to the cauliflower mosaic virus (CAMV) 35S promoter, the maize ubiquitin promoter, which can be used alone or in combination with other plant promoters; in addition, when constructing the plant expression vector using the gene of the present application, enhancers can also be used, including translation enhancers or transcription enhancers, and these enhancer regions can be the ATG start codon or the adjacent region start codon, etc., but must be the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The source of the translation control signal and the start codon is wide and can be natural or synthetic. The translation initiation region can be 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 to produce color changes or luminescent compounds (GUS gene, luciferase gene, etc.), antibiotic markers with resistance (gentamicin marker, kanamycin marker, etc.), or chemical reagent-resistant marker genes (e.g., herbicide-resistant genes), etc. For the safety of transgenic plants, no selective marker gene can be added and the transformed plants can be directly screened under stress conditions.
[0052] The present application also provides a method for changing the salt tolerance and root length of plants, which comprises the following steps M or P:
[0053] The step M is to enhance, increase or up-regulate the activity and / or content of the protein as described above in the plant of interest, or / and, to enhance, increase or up-regulate the expression of the gene encoding the protein as described above, so as to increase the salt tolerance and root length of the plant.
[0054] The method comprises a step P of inhibiting or reducing or silencing the activity and / or content of the protein as described above in the plant of interest, or / and, inhibiting or reducing or silencing the expression of the gene encoding the protein as described above, so as to decrease the salt tolerance and root length of the plant.
[0055] In the above method, the expression and / or activity of the gene encoding the protein OsPL3 in the plant of interest can be reduced or inactivated by gene mutation, gene knockout, gene editing or gene knockdown technology.
[0056] The present application also provides a method for breeding a plant with improved salt tolerance and elongated root, which comprises up-regulating or enhancing or increasing the expression of the gene encoding the protein as described above in the plant of interest, and / or the activity and / or content of the protein, so as to obtain a plant with improved salt tolerance and elongated root, wherein the salt tolerance of the plant with improved salt tolerance is higher than that of the plant of interest, and the root length of the plant with elongated root is longer than that of the plant of interest.
[0057] In a specific embodiment, the up-regulation or enhancement or increase of the expression of the gene encoding the protein as described above in the plant comprises introducing the nucleic acid molecule, expression cassette or recombinant vector as described above into the plant of interest, so as to obtain a plant with high salt tolerance and long root.
[0058] The present application also provides a method for breeding a plant with reduced salt tolerance and shortened root, which comprises inhibiting or reducing or silencing the expression of the gene encoding the protein as described above in the plant of interest, and / or the activity and / or content of the protein, so as to obtain a plant with reduced salt tolerance and shortened root, wherein the salt tolerance of the plant with reduced salt tolerance is lower than that of the plant of interest, and the root length of the plant with shortened root is shorter than that of the plant of interest.
[0059] In a specific embodiment, the inhibition or reduction or silencing of the expression of the gene encoding the protein as described above in the plant comprises introducing the nucleic acid molecule, expression cassette or recombinant vector as described above into the plant of interest, so as to obtain a plant with reduced salt tolerance and shortened root.
[0060] In this application, the breeding purposes include breeding a plant with high salt tolerance and elongated root, and the breeding purposes also include breeding a plant with low salt tolerance and shortened root.
[0061] The high salt tolerance and root length plant has improved salt tolerance and elongated root length compared with the plant of interest. The low salt tolerance and root length plant has reduced salt tolerance and shortened root length compared with the plant of interest.
[0062] Herein, the improved salt tolerance can be improved survival rate. The survival rate can be survival rate at seedling stage.
[0063] Herein, the rice can be rice ZH11.
[0064] Herein, the recombinant microorganism can be Agrobacterium EHA105.
[0065] The protein and / or the biological material described in the foregoing also belong to the scope of protection of the present application.
[0066] In the above application or method, the plant can be any one of the following:
[0067] N1) monocotyledonous plants;
[0068] N2) plants of the order Poales;
[0069] N3) plants of the family Poaceae;
[0070] N4) plants of the genus Oryza;
[0071] N5) rice.
[0072] The present application discloses the regulatory effect of OsPL3-OE protein and its encoding gene on the stress tolerance, growth performance and production performance of plants. The present application protects the application of OsPL3 protein in regulating the stress tolerance of plants. The regulation is positive regulation, that is, the content of OsPL3 protein is increased, and the salt tolerance of the plant is enhanced. The present application also protects the application of OsPL3 protein in regulating the growth performance and / or production performance of plants. The present application can be used for improving the stress tolerance of plants, and has great application and promotion value for plant breeding, especially rice breeding. BRIEF DESCRIPTION OF DRAWINGS
[0073] Figure 1 The schematic diagram of the structure of the construction vector for constructing the overexpression strain.
[0074] Figure 2 The expression level of OsPL3 gene of the overexpression strain.
[0075] Figure 3 OsPL3 positively regulates the root growth and development of rice. Wherein A is the phenotype of seedlings grown for one week of wild type and OsPL3-OE; B is the root length statistics of wild type and OsPL3-OE.
[0076] Figure 4The statistical chart of phenotype and survival rate of wild type and OsPL3 overexpression lines before and after salt stress treatment. A is the chart of phenotype of wild type and OsPL3 overexpression lines before and after salt stress treatment; B is the statistical chart of survival rate of wild type and OsPL3 overexpression lines after salt stress treatment. DETAILED DESCRIPTION
[0077] The application will be further described in conjunction with the specific embodiments. The examples given are only to illustrate the application, and are not intended to limit the scope of the application. The examples provided below can serve as a guide for further improvement by those skilled in the art, and do not in any way constitute a limitation on the application.
[0078] The experimental methods in the following examples are all routine methods, unless otherwise specified, which are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained commercially.
[0079] The quantitative tests in the following examples, unless otherwise specified, are all set up with three repeated experiments, and the results are taken as the average value.
[0080] The 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 biological material can be obtained from the applicant, and can only be used for repeating the experiments of the application, 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, W. A. Tao, X.-J. He, H. Zhang, R. A. Bressan, J. Du, J. K. Zhu, Arabidopsis AGDP1 links H3K9me2 to DNA methylation in heterochromatin. Nat. Commun. 9, 4547 (2018). doi:10.1038 / s41467-018-06965-wMedlin. The biological material is available to the public from the applicant, which is only used for repeating the experiments of the present application and cannot be used for other purposes.
[0082] Rice ZH11 in the following examples has been described 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 biological material is available to the public from the applicant, which is only used for repeating the experiments of the present application and cannot be used for other purposes.
[0083] SPSS11.5 statistical software was used to process the data in the following examples, and the experimental results were expressed as mean ± standard deviation. One-way ANOVA test was used, P<0.05 (*) indicated significant difference, P<0.01 (**) indicated extremely significant difference, and P<0.001 (***) indicated extremely significant difference.
[0084] Example 1, obtaining of rice OsPL3 gene
[0085] The leaf RNA of rice variety (Nip) was extracted and reverse transcribed into cDNA. The cDNA was used as a template, and MaxSuper-Fidelity DNA Polymerase (item number: P505-d1, Vazyme) was used for PCR amplification with primers OsPL3-F: 5'-ATGAGATCTGTCTCTGAGCT-3'; OsPL3-R: 5'-AAAACGAAGCAGCTCTCTTT-3', to obtain the amplification product (i.e. the coding region of OsPL3 gene).
[0086] The coding sequence (CDS) of the OsPL3 gene in rice variety Nip is SEQ ID No. 2, which encodes the OsPL3 with the amino acid sequence shown in SEQ ID No. 1. The gene encoding the OsPL3 protein in the genomic DNA of rice Nip is shown in SEQ ID No. 3 of the sequence listing. The first exon is 1230-1551, the second exon is 1929-2063, the third exon is 2131-4045, the fourth exon is 4692-4807, the fifth exon is 4924-5015, and the sixth exon is 5168-5275 of SEQ ID No. 3.
[0087] Example 2, construction of recombinant plasmid pSuper1300-OsPL3-GFP
[0088] The coding region sequence of OsPL3 was amplified by PCR using the cDNA of Nip as the template and primers OsPL3-F / OsPL3-R, and the target gene fragment of OsPL3 was recovered by gel.
[0089] After the target gene and the vector were respectively digested by the same restriction enzyme (XbaI), linked, and transformed into E. coli, pSuper1300-OsPL3-GFP overexpression vector was obtained through one-generation sequencing verification and sequence alignment. The structure of the recombinant plasmid pSuper1300-OsPL3-GFP is shown in Figure 1 .
[0090] The recombinant vector pSuper1300-OsPL3-GFP is a DNA molecule (OsPL3 coding gene) with the nucleotide sequence of SEQ ID No. 2, which replaces the small fragment of the pSuper1300 vector restriction enzyme XbaI enzyme recognition site, and the other sequences of the pSuper1300 vector remain unchanged, to obtain the recombinant vector. The full-plasmid sequencing showed that the recombinant plasmid pSuper1300-OsPL3-GFP is shown in SEQ ID No. 4.
[0091] Example 3, obtaining of transgenic rice
[0092] The recombinant plasmid pSuper1300-OsPL3-GFP obtained in Example 2 was introduced into Agrobacterium tumefaciens EHA105 to obtain a recombinant Agrobacterium. The embryonic callus of rice ZH11 was genetically transformed by the Agrobacterium dipping method using the recombinant Agrobacterium, and then resistant callus was screened (resistance screening used 100 mg / L hygromycin), then differentiation and 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 mature seeds of the plant, remove the hulls, and pick the full, clean, and sterile seeds for sterilization.
[0095] (2) Seed the sterilized seeds on an induction medium, and cultivate at 28°C in the dark for about 14 days to select the callus with good appearance and growth.
[0096] (3) Take the recombinant vector pSuper1300-OsPL3-GFP constructed in Example 2, introduce it into Agrobacterium tumefaciens EHA105, and obtain a recombinant bacterium, which is named EHA105 / pSuper1300-OsPL3-GFP.
[0097] (4) Take the recombinant bacterium obtained in step (3), resuspend the bacterium in an infection medium, and obtain a bacterium suspension of EHA105 / pSuper1300-OsPL3-GFP.
[0098] (5) Soak the Nip callus of step (2) in the bacterium suspension of EHA105 / pSuper1300-OsPL3-GFP prepared in step (4) for 20 min. After the infection, pour off the bacterium suspension, dry the callus with sterile filter paper, and then place it on a co-cultivation medium added with acetosyringone and glucose, and cultivate at 28°C in the dark for 50-55 h.
[0099] (6) After step (5) is completed, select the callus without obvious Agrobacterium on the surface, and move it to an antibiotic medium added with cephalosporin, and cultivate at 28°C in the dark for 3-4 days.
[0100] (7) Move the callus after the cultivation to a selection medium added with hygromycin and cephalosporin (the concentration of hygromycin is 100 mg / L and the concentration of cephalosporin is 100 mg / L), and cultivate at 28°C in the dark for 30 days, and subculture every 10 days.
[0101] (8) After step (7) is completed, take the fresh hygromycin-resistant callus, inoculate it in a pre-regeneration medium, cultivate at 28°C in the dark for 7 days, then place it in a light cultivation room (12 h light / 12 h dark) for continuous cultivation for 7 days, and then move it to a regeneration medium for continuous light cultivation until the regenerated plant grows, and obtain the OsPL3-OE transgenic plant. The medium used for genetic transformation and the formula are as follows: the formula of the induction medium and the differentiation medium is MS medium.
[0102] The transgenic plant obtained by using the recombinant vector pSuper1300-OsPL3-GFP is recorded as the OsPL3 transgenic plant.
[0103] Example 4, Identification of the transgenic rice and detection of the gene expression level
[0104] 1. Identification of OsPL3 transgenic rice
[0105] Tested plants: rice Nip (CK) and OsPL3 transgenic plants obtained in Example 3.
[0106] Genomic DNA of the tested plants was extracted, and PCR amplification was performed using the primer pair consisting of primers OsPL3-F / OsPL3-R, with the genomic DNA as a template, and using the pSuper1300-OsPL3-GFP plasmid as a positive control (V) and the receptor variety Nip as a negative control (CK). Then the obtained products were 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 germinated and seedlings were grown in a greenhouse (the timing started from the emergence of green, and the cultivation lasted for 3 weeks), the seedling materials were quickly frozen in liquid nitrogen, RNA was extracted (TIANGEN kit, item number: DP419), reverse transcription was performed (Novizen kit, item number: R333-01), and gene expression was detected by fluorescence quantitative PCR. The detection primers were as follows:
[0109] OsPL3-RT-F: 5'-GTCCGTGTCTGCTGTGAATC-3';
[0110] OsPL3-RT-R: 5'-GCTTTTGTACTTGGGCAGCT-3'.
[0111] It can be seen from Table 2 that the expression levels of different OsPL3 transgenic rice genes were significantly increased. Figure 2
[0112] Example 5, Plant root phenotype and salt tolerance performance research
[0113] Tested seeds: rice Nippanese seed (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. The test rice seeds were germinated and seedlings were grown in a greenhouse (the timing started from the emergence of green, and the cultivation lasted for 1 week), 1-week-old seedlings were obtained, and photographs were taken; the root length was counted.
[0115] It can be seen from Table 3 that the root length of OsPL3-OE was significantly longer than that of the wild type. Figure 3
[0116] 2. The test plants were cultured under parallel conditions, specifically: the test rice seeds were germinated and seedlings were grown in a greenhouse (the time was counted from the beginning of germination, and the culture lasted for 3 weeks), and 3-week-old seedlings were obtained and photographed; the 3-week-old seedlings were treated with 150 mM NaCl for 5 days, and the phenotype and survival rate were counted and photographed Figure 4 . The greenhouse conditions were: 28°C, 10 hours of light / 14 hours of darkness.
[0117] From Figure 4 A and B, it can be seen that the survival rate of the OsPL3-OE seedlings was obviously increased by about 1 fold compared with the wild type after salt stress treatment, indicating that OsPL3 positively regulates the salt stress response of rice.
[0118] The present application has been described in detail. For those skilled in the art, without departing from the purpose and scope of the present application, and without unnecessary experiments, the present application can be implemented in a wide range under the same parameters, concentrations and conditions. Although the present application gives a special example, it should be understood that the present application can be further improved. In short, according to the principle of the present application, this application intends to include any change, use or improvement of the present application, including the changes made by the conventional techniques known in the art, which deviates from the range disclosed in the present application.
Claims
1. The use of a substance that enhances the expression of a protein-coding gene or the activity or content of said protein in any of the following; 1) Application in improving the salt tolerance and root length of rice; 2) Application in the preparation of products that improve the salt tolerance and root length of rice; 3) Application in cultivating rice varieties with salt tolerance and longer root length; 4) Application in the preparation of rice products with salt tolerance and longer root length; The protein is any of the following: a1) A protein with the amino acid sequence SEQ ID No. 1; a2) The fusion protein obtained by attaching a tag to the end of the protein defined in a1).
2. The application according to claim 1, characterized in that, The protein is derived from rice.
3. The application 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 of claim 1 or 2, and the biological material is any one of the following: c1) The 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); wherein the recombinant microorganism is recombinant Agrobacterium.
4. The application according to claim 3, characterized in that: c1) The nucleic acid molecule is any of the following DNA molecules. 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.
5. A method for altering the salt tolerance and root length of rice, characterized in that, The method includes enhancing, increasing, or upregulating the activity and / or content of the protein described in claim 1 or 2 in the target rice, or / and enhancing, increasing, or upregulating the expression level of the gene encoding the protein described in claim 1 or 2, to improve the salt tolerance and root length of rice.
6. A method for cultivating rice with improved salt tolerance and root elongation, characterized in that, This includes enhancing, increasing, or upregulating the expression level of the gene encoding the protein described in claim 1 or 2 in the target rice, and / or, the activity and / or content of the protein to obtain rice with improved salt tolerance and root elongation, wherein the salt tolerance of the rice with improved salt tolerance and root elongation is higher than that of the target rice, and the roots are longer than those of the target rice.
7. The method according to claim 6, characterized in that, The enhancement, improvement, or upregulation of the expression of the gene encoding the protein of claim 1 or 2 in rice comprises introducing the nucleic acid molecule of claim 4c1), the expression cassette of claim 4c2), or the recombinant vector of claim 4c3) into the target rice to obtain rice with improved salt tolerance and longer root length.