CN114276427B - Application of OsFTL1 and its coding gene in shortening the heading date of rice - Google Patents

Abstract

The invention discloses the application of OsFTL1 and its coding gene in shortening the heading period of rice. OsFTL1, its sequence is sequence 1 in the sequence listing, and its coding gene sequence is sequence 2 in the sequence listing. Experiments have proved that OsFTL1 and related biological materials of the present invention can promote early heading of rice, and the heading date of OsFTL1 gene-transferred rice is significantly earlier than that of wild-type rice, and the transgenic rice has already headed only 45-47 days after sowing. At this time, the wild type is still in the tillering stage, and correspondingly, the heading date of the wild type is 116-118 days after sowing. This shows that the heading date of the transgenic rice can be shortened by about 71 days compared with the wild type, and the grain filling maturity period of the rice is correspondingly earlier. It shows that OsFTL1 and its coding gene can regulate the heading date of rice.

Description

Application of OsFTL1 and its coding gene in shortening the heading date of rice

technical field

The invention relates to the application of OsFTL1 and its coding gene in shortening the heading period of rice in the field of biotechnology.

Background technique

Rice (Oryza sativa L.) is one of the three major food crops in the world, and more than half of the world's population uses rice as a staple food. As the world's population continues to grow, while climate change and human activities have reduced the area of land suitable for cultivation, how to increase rice yield is still the focus of crop research today. In agricultural production, the growth period of crops plays a decisive role in crop yield, among which the heading date of rice is one of the most important agronomic traits that determine the yield and quality of rice. The appropriate heading date is very important for rice varieties to adapt to different ecological regions and different planting conditions. Seasons play a key role. For a long time, breeding early-maturing and high-yielding rice varieties has been the focus and focus of breeders' research. The analysis of the regulation pathway of heading date and the study of the molecular mechanism of heading date genes are of great importance for crop breeding, variety improvement and variety promotion. meaning.

The main influencing factors of heading date include variety differences, photoperiod and farming methods, etc. The difference in heading date of different rice varieties directly affects the cultivation area and seasonal adaptability of varieties, and photoperiod is one of the most important environmental factors to regulate heading date. In recent years, at least 14 genes involved in the regulation of rice heading date have been found through QTL mapping research, and the molecular regulatory network of rice heading date has been basically clarified. It is known that there are two main regulatory pathways for rice heading date, including the Hd1-dependent OsGI-Hd1-Hd3a pathway and the Ehd1-dependent Ghd7-Ehd1-RFT1/Hd3a pathway. These two pathways are functional under short-day conditions. Redundant, but antagonistic under long-day light. Plants collect flowering signals from Hd1 and Ehd1 and transduce them to florigen genes Hd3a and RFT1 to promote flowering in rice under long- and short-day conditions, respectively. Among them, Hd1 is the homologous gene of Arabidopsis flowering key gene CONSTANS (CO) in rice, which is regulated by OsGI, promotes flowering by activating the expression of Hd3a gene under short-day light, and inhibits the expression delay of Hd3a gene under long-day light Flowering, this pathway is homologous to the GI-CO-FT pathway in Arabidopsis. The Ehd1 gene in the other pathway is highly conserved in rice, encoding a B-type response regulator, which is regulated by Ghd7 to inhibit flowering, and can promote flowering by activating Hd3a and RFT1 genes, which is a unique regulatory pathway in rice. The Ehd1 gene is the center of various signal transduction. Under short-day light, Ehd1 gene can promote the early heading of rice by inducing the expression of FT-Like genes, while under long-day light, its expression is regulated by multiple regulatory factors, among which positive regulators include Ehd2, Ehd4, MADS50 and MADS51, negative regulators include Ghd7, Ghd8, OsLFL1, etc.

At present, although some progress has been made in the research on the regulation pathways of rice heading date and related functional genes, and the important key pathways have been basically understood, the detailed flowering regulation mechanism is still unclear, and many key genes that regulate heading date are still unknown. is unknown. Therefore, the excavation and molecular mechanism research of functional genes controlling rice heading date will help us to further improve the understanding of the regulation mechanism of rice heading date, and based on this, we can take targeted measures to improve the heading date of rice varieties and improve the rice varieties. Regional adaptability and seasonal adaptability, further ensuring the improvement of yield and quality, have important theoretical significance and application value for solving the long-standing contradiction in agricultural production that "premature maturity and high yield are difficult to balance".

Contents of the invention

The technical problem to be solved by the present invention is how to shorten the heading period (flowering period) of the plant or prolong the heading period (flowering period) of the plant.

In order to solve the above-mentioned technical problems, the present invention firstly provides any of the following applications of proteins or substances that regulate the activity or content of the proteins:

D1) regulating plant flowering time;

D2) preparing products for regulating the flowering time of plants;

D3) plant breeding;

The protein (whose name is OsFTL1) is A1), A2), A3) or A4) as follows:

A1) the amino acid sequence is the protein of sequence 1;

A2) A protein having the same function as the amino acid sequence shown in Sequence 1 in the sequence listing through substitution and/or deletion and/or addition of one or several amino acid residues;

A3) A protein derived from corn, sorghum, millet, goatgrass, Brachypodium distachyon or wheat that has 75% or more identity to Sequence 1 and has the same function as the protein described in A1);

A4) A fusion protein obtained by linking a tag at the N-terminal or/and C-terminal of A1) or A2) or A3).

In order to make the protein in A1) easy to purify, the amino-terminal or carboxy-terminal of the protein consisting of the amino acid sequence shown in Sequence 1 in the sequence listing can be attached with the tags shown in the table below.

Table: Sequence of Labels

Label Residues sequence Poly-Arg 5-6 (usually 5) RRRRR Poly-His 2-10 (usually 6) HHHHHH FLAG 8 DYKDDDDK Strep-tag II 8 WSHPQFEK c-myc 10 EQKLISEEDL

The protein in A2) above is a protein having 75% or more identity to the amino acid sequence of the protein shown in Sequence 1 and having the same function. Said having 75% or more identity means having 75%, having 80%, having 85%, having 90%, having 95%, having 96%, having 97%, having 98% or having 99% identity .

The protein in the above A2) can be synthesized artificially, or its coding gene can be synthesized first, and then obtained by biological expression.

The gene encoding the protein in the above A2) can be deleted by deleting one or several amino acid residue codons in the DNA sequence shown in Sequence 2, and/or performing missense mutations of one or several base pairs, and/ Or it can be obtained by connecting the coding sequence of the tag shown in the above table at its 5' end and/or 3' end. Wherein, the DNA molecule shown in sequence 2 encodes the protein shown in sequence 1.

The present invention also provides any of the following applications of biological materials related to OsFTL1:

D1) regulating plant flowering time;

D2) preparing products for regulating the flowering time of plants;

D3) plant breeding;

The biological material is any one of the following B1) to B7):

B1) a nucleic acid molecule encoding OsFTL1;

B2) an expression cassette containing the nucleic acid molecule of B1);

B3) a recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2);

B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing a recombinant vector described in B3);

B5) a transgenic plant cell line containing the nucleic acid molecule described in B1), or a transgenic plant cell line containing the expression cassette described in B2);

B6) a transgenic plant tissue containing the nucleic acid molecule described in B1), or a transgenic plant tissue containing the expression cassette described in B2);

B7) A transgenic plant organ containing the nucleic acid molecule described in B1), or a transgenic plant organ containing the expression cassette described in B2).

In the above application, the nucleic acid molecule described in B1) can be as follows b11) or b12) or b13) or b14) or b15):

b11) The coding sequence is a cDNA molecule or a DNA molecule of sequence 2 in the sequence listing;

b12) cDNA molecules or DNA molecules shown in sequence 2 in the sequence listing;

b13) DNA molecules shown in sequence 3 in the sequence listing;

b14) has 75% or more identity with the nucleotide sequence defined by b11) or b12) or b13), and encodes a cDNA molecule or DNA molecule of OsFTL1;

b15) A cDNA molecule or a DNA molecule that hybridizes to the nucleotide sequence defined by b11) or b12) or b13) or b14) under stringent conditions and encodes OsFTL1.

Wherein, the nucleic acid molecule can be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule can also be RNA, such as mRNA or hnRNA.

Those skilled in the art can easily use known methods, such as directed evolution and point mutation methods, to mutate the nucleotide sequence encoding the OsFTL1 protein of the present invention. Those artificially modified nucleotides with 75% or higher identity to the nucleotide sequence of the OsFTL1 protein isolated in the present invention, as long as they encode the OsFTL1 protein and have the function of the OsFTL1 protein, are all derived from the core of the present invention. Nucleotide sequence and is equivalent to the sequence of the present invention.

The term "identity" as used herein refers to sequence similarity to a native nucleic acid sequence. "Identity" includes 75% or higher, or 85% or higher, or 90% or higher, or 95% or higher, of the nucleotide sequence of the protein composed of the amino acid sequence shown in the coding sequence 1 of the present invention. Nucleotide sequences of higher identity. Identity can be assessed visually or with computer software. Using computer software, identity between two or more sequences can be expressed as a percentage (%), which can be used to evaluate the identity between related sequences.

In the above application, the stringent conditions may be as follows: 50°C, hybridization in a mixed solution of 7% sodium dodecyl sulfate (SDS), 0.5M NaPO ₄ and 1mM EDTA, at 50°C, 2×SSC, 0.1% Rinse in SDS; can also be: 50°C, hybridize in a mixed solution of 7% SDS, 0.5M NaPO ₄ and 1mM EDTA, rinse in 50°C, 1×SSC, 0.1% SDS; can also be: 50°C, Hybridize in a mixed solution of 7% SDS, 0.5M NaPO ₄ and 1mM EDTA, rinse at 50°C in 0.5×SSC, 0.1% SDS; also: 50°C, in 7% SDS, 0.5M NaPO ₄ and 1mM Hybridize in a mixed solution of EDTA, rinse at 50°C, 0.1×SSC, 0.1% SDS; also: 50°C, hybridize in a mixed solution of 7% SDS, 0.5M NaPO ₄ and 1mM EDTA, at 65°C, Rinse in 0.1×SSC, 0.1% SDS; alternatively: in 6×SSC, 0.5% SDS solution, hybridize at 65°C, then use 2×SSC, 0.1% SDS and 1×SSC, 0.1% SDS each Wash the membrane once; it can also be: in 2×SSC, 0.1% SDS solution, hybridize at 68°C and wash the membrane twice, each time for 5 minutes, then in 0.5×SSC, 0.1% SDS solution, at 68°C Hybridize and wash the membrane twice for 15 minutes each time; alternatively: in a solution of 0.1×SSPE (or 0.1×SSC) and 0.1% SDS, hybridize and wash the membrane at 65°C.

The identity of 75% or more may be 80%, 85%, 90% or more.

In the above-mentioned application, the expression cassette (OsFTL1 gene expression cassette) described in B2) containing the nucleic acid molecule encoding the OsFTL1 protein refers to the DNA capable of expressing the OsFTL1 protein in the host cell. A terminator that terminates the transcription of the OsFTL1 gene may also be included. Further, the expression cassette may also include an enhancer sequence. Promoters that can be used in the present invention include, but are not limited to: constitutive promoters, tissue, organ and development specific promoters, and inducible promoters. Examples of promoters include, but are not limited to: the constitutive promoter 35S of cauliflower mosaic virus; the wound-inducible promoter from tomato, leucine aminopeptidase ("LAP", Chao et al. (1999) PlantPhysiol 120:979 -992); chemically inducible promoter from tobacco, pathogenesis-related 1 (PR1) (induced by salicylic acid and BTH (benzothiadiazole-7-thiohydroxy acid S-methyl ester)); tomato protease Inhibitor II promoter (PIN2) or LAP promoter (both inducible with methyl jasmonate); heat shock promoter (US Patent 5,187,267); tetracycline-inducible promoter (US Patent 5,057,422); seed-specific Genetic promoters, such as millet seed-specific promoter pF128 (CN101063139B (Chinese patent 200710099169.7)), seed storage protein-specific promoters (for example, the promoters of phaseolin, napin, oleosin and soybean beta conglycin (Beachy et al. ( 1985) EMBO J. 4:3047-3053)). They can be used alone or in combination with other plant promoters. All references cited herein are cited in their entirety. Suitable transcription terminators include, but are not limited to: Agrobacterium nopaline synthase terminator (NOS terminator), cauliflower mosaic virus CaMV 35S terminator, tml terminator, pea rbcS E9 terminator and nopaline and octopine Synthase terminators (see, e.g.: Odell et al. (1985) Nature 313:810; Rosenberg et al. (1987) Gene, 56:125; ^Guerineau et al. (1991) Mol. Gen. Genet, 262:141; Proudfoot (1991) Cell, 64:671; Sanfacon et al. Genes Dev., 5:141; Mogen et al. (1990) Plant Cell, 2:1261; Munroe et al. (1990) Gene, 91:151; Ballad et al. (1989) ) Nucleic Acids Res. 17:7891; Joshi et al. (1987) Nucleic Acids Res., 15:9627).

The existing expression vector can be used to construct the recombinant vector containing the OsFTL1 gene expression cassette. The plant expression vectors include binary Agrobacterium vectors and vectors that can be used for plant microprojectile bombardment and the like. Such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa, PSN1301 or pCAMBIA1391-Xb (CAMBIA Company), etc. The plant expression vector may also include the 3' untranslated region of the foreign gene, that is, the polyadenylation signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylic acid signal can guide polyadenylic acid to be added to the 3' end of the mRNA precursor, such as Agrobacterium crown gall tumor induction (Ti) plasmid gene (such as nopaline synthase gene Nos), plant gene (such as soybean The untranslated region transcribed at the 3′ end of the storage protein gene) has similar functions. When using the gene of the present invention to construct plant expression vectors, enhancers can also be used, including translation enhancers or transcription enhancers, and these enhancer regions can be ATG initiation codons or adjacent region initiation codons, etc. The reading frames of the sequences are identical to ensure correct translation of the entire sequence. The sources of the translation control signals and initiation codons are extensive and can be natural or synthetic. The translation initiation region can be from a transcription initiation region or a structural gene. 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 (GUS gene, luciferase gene, etc.) genes, etc.), antibiotic marker genes (such as the nptII gene that confers resistance to kanamycin and related antibiotics, the bar gene that confers resistance to the herbicide phosphinothricin, and the hph gene that confers resistance to the antibiotic hygromycin , and the dhfr gene that confers resistance to methotrexate, the EPSPS gene that confers resistance to glyphosate) or the chemical resistance marker gene (such as the herbicide resistance gene), the mannose-6- that provides the ability to metabolize mannose Phosphate isomerase gene. Considering the safety of the transgenic plants, the transformed plants can be screened directly by adversity without adding any selectable marker gene.

In the above application, the vector can be a plasmid, cosmid, phage or viral vector. Specifically, the plasmid can be a pTCK303 vector.

B3) The recombinant vector can specifically be pTCK303-OsFTL1. The pTCK303-OsFTL1 is a recombinant vector obtained by replacing the DNA fragment between the BamHI and SacI recognition sequences of the pTCK303 vector with the OsFTL1 gene shown in Sequence 2. The pTCK303-OsFTL1 can express the protein encoded by the OsFTL1 gene (ie the OsFTL1 protein shown in Sequence 1) under the drive of the maize Ubiqutin (UBI) promoter.

In the above applications, the microorganisms can be yeast, bacteria, algae or fungi. Wherein, the bacteria can be Agrobacterium, such as Agrobacterium rhizogenes EHA105.

In the above applications, the transgenic plant cell lines, transgenic plant tissues and transgenic plant organs do not include propagation materials.

The present invention also provides a method for cultivating photosynthesis-enhanced plants, the method comprising expressing OsFTL1 in recipient plants, or increasing the content or activity of OsFTL1 in recipient plants to obtain target plants with enhanced photosynthesis.

The above method can be realized by introducing the coding gene of OsFTL1 into the recipient plant and expressing the coding gene.

In the above method, the encoding gene may be the nucleic acid molecule described in B1).

In the above method, the gene encoding OsFTL1 can be modified as follows first, and then introduced into the recipient plant to achieve a better expression effect:

1) Modify and optimize according to actual needs, so that the gene can be expressed efficiently; for example, according to the codon preferred by the recipient plant, its codon can be changed while maintaining the amino acid sequence of the gene encoding OsFTL1 according to the present invention to conform to Plant preference; during the optimization process, it is best to maintain a certain GC content in the optimized coding sequence, so as to best achieve high-level expression of the introduced gene in plants, where the GC content can be 35% or more than 45% , more than 50% or more than about 60%;

2) modifying the gene sequence adjacent to the starting methionine to allow efficient initiation of translation; for example, using sequences known to be effective in plants for modification;

3) Linking with various plant-expressed promoters to facilitate its expression in plants; said promoters may include constitutive, inducible, temporally regulated, developmentally regulated, chemically regulated, tissue-preferred and tissue-specific promoters ; the choice of promoter will vary with the temporal and spatial requirements of expression, and also depends on the target species; e.g. a tissue or organ-specific expression promoter, depending on what stage of development the recipient is desired; although proven source Many promoters for dicots are functional in monocots and vice versa, but ideally, dicot promoters are chosen for expression in dicots and monocot promoters are used for Expression in monocots;

4) Linking with suitable transcription terminators can also improve the expression efficiency of the gene of the present invention; for example, tml derived from CaMV, E9 derived from rbcS; any available terminators known to work in plants can be combined with The gene of the present invention is connected;

5) Introduce enhancer sequences, such as intron sequences (eg derived from Adhl and bronze) and viral leader sequences (eg derived from TMV, MCMV and AMV).

The coding gene of OsFTL1 can be introduced into recipient plants by means of a recombinant expression vector containing the coding gene of OsFTL1. The recombinant expression vector can specifically be the pTCK303-OsFTL1.

The recombinant expression vector can be introduced into plant cells by conventional biotechnological methods such as Ti plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation (Weissbach, 1998, Method for Plant Molecular Biology VIII, Academy Press, New York, pp.411-463; Geiserson and Corey, 1998, Plant Molecular Biology (2nd Edition).).

The target plant is understood to include not only the first-generation plant whose OsFTL1 protein or its coding gene has been changed, but also its progeny. For the desired plant, the gene can be propagated in the species, or can be transferred into other varieties of the same species, including commercial varieties in particular, by conventional breeding techniques. The target plants include seeds, callus, whole plants and cells.

The present invention also provides a product with the function of regulating plant flowering time, said product containing OsFTL1 or said biological material.

In the above, the plant may be M1) or M2) or M3):

M1) monocot or dicot;

M2) grasses, crucifers or legumes;

M3) Rice, wheat, corn, Arabidopsis, rape or soybean.

In the above, the flowering time can be reflected in the heading stage.

OsFTL1 or the biological material also belongs to the protection scope of the present invention.

Experiments have proved that OsFTL1 and related biological materials of the present invention can promote early heading of rice. The heading date of OsFTL1 gene-transformed rice is significantly earlier than that of wild-type rice, and the transgenic rice has already headed only 45-47 days after sowing. At this time, the wild type is still in the tillering stage, and correspondingly, the heading stage of the wild type is 116-118 days after sowing. This shows that the heading date of the transgenic rice can be shortened by about 71 days compared with the wild type, and the grain filling maturity period of the rice is correspondingly earlier. It shows that OsFTL1 and its coding gene can regulate the heading date of rice.

Description of drawings

Figure 1 shows the sequence alignment results.

Fig. 2 is the detection result of relative expression level of FTL1 gene in OsFTL1 gene transgenic rice.

Fig. 3 is a photo of wild type and OsFTL1 transgenic rice. A- OsFTL1 gene transgenic rice heads earlier than wild type; B-OsFTL1 gene transgenic rice mature stage phenotype.

Figure 4 shows the heading dates of wild-type and OsFTL1-transgenic rice under long-day conditions in Beijing (A) and short-day conditions in Hainan (B).

In Fig. 3-4, ** indicates that compared with WT, the difference reached extremely significant level, p<0.01.

detailed description

The present invention will be further described in detail below in conjunction with specific embodiments, and the given examples are only for clarifying the present invention, not for limiting the scope of the present invention. The examples provided below can be used as a guideline for those skilled in the art to make further improvements, and are not intended to limit the present invention in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. Materials, reagents, instruments, etc. used in the following examples can be obtained from commercial sources unless otherwise specified. Quantitative experiments in the following examples were all set up to repeat the experiments three times, and the results were averaged. In the following examples, unless otherwise specified, the first position of each nucleotide sequence in the sequence listing is the 5' terminal nucleotide of the corresponding DNA/RNA, and the last position is the 3' terminal nucleotide of the corresponding DNA/RNA glycosides.

The pTCK303 vector in the following examples (Zhang, H., Zhang, J., Yan, J., Gou, F., Mao, Y., Tang, G., Botella, J.R., & Zhu, J.K. (2017). Short tandem target mimic rice lines cover functions of miRNAs in regulating important agronomic traits. Proceedings of the National Academy of Sciences of the United States of America, 114(20), 5277–5282.

https://doi.org/10.1073/pnas.1703752114), the public can obtain the biological material from the applicant, and the biological material is only used for repeating related experiments of the present invention, and cannot be used for other purposes.

Example 1, OsFTL1 can promote the early heading date of rice

This example provides a protein derived from Nipponbare rice that can promote the heading date. The name of the protein is OsFTL1, and its sequence is sequence 1 in the sequence table. In Nipponbare, the coding gene sequence of OsFTL1 is sequence 2, and the genome sequence for sequence 3.

Comparing the sequences of rice OsFTL1 with homologous proteins in other plants, it was found that its identities with corn, sorghum, millet, goat grass, wheat, and Brachypodium distachyon were 94.15%, 93.57%, 93.57%, and 92.40%, respectively. %, 92.40%, 92.98% (Figure 1).

1. Construction of recombinant vector

Artificially synthesize the OsFTL1 gene shown in sequence 2 in the sequence listing, replace the DNA fragment between the BamHI and SacI recognition sequences of the pTCK303 vector with the OsFTL1 gene shown in sequence 2, keep other sequences unchanged, obtain the recombinant vector, and name it as pTCK303-OsFTL1. pTCK303-OsFTL1 can express the protein encoded by the OsFTL1 gene (ie the OsFTL1 protein shown in Sequence 1) under the drive of maize Ubiqutin (UBI) promoter.

2. Construction of transgenic plants

The embryogenic callus was induced by disinfecting the mature seeds of the japonica rice variety Nipponbare, and the pTCK303-OsFTL1 obtained in step 1 was introduced into Agrobacterium EHA105, and the callus was infected by the method of Agrobacterium-mediated genetic transformation of rice Co-cultivate, use resistance screening to obtain transgenic plants, and the screened transgenic rice is OsFTL1 transgenic rice.

Using the japonica rice variety Nipponbare (WT) as a control, the relative expression level of OsFTL1 gene at the RNA level in OsFTL1 transgenic rice was detected by qRT-PCR. 3'; the internal reference gene is rice Ubiqutin gene, and the internal reference gene primers are: 5'-AAGAAGCTGAAGCATCCAGC-3', 5'-CCAGGACAAGATGATCTGCC-3'.

The results showed that the relative expression levels of the OsFTL1 gene in the three OsFTL1 transgenic rice lines (OsFTL1-OE1, OsFTL1-OE2 and OsFTL1-OE3) were significantly higher than those in the wild type (WT), and all three lines were overexpressed OsFTL1 rice material (Fig. 2).

3. The heading date of rice transgenic for OsFTL1 is advanced

Detect the heading date of rice, the rice to be tested: wild type Nipponbare rice (WT), rice overexpressing OsFTL1 (OsFTL1-OE1, OsFTL1-OE2 and OsFTL1-OE3).

The heading date of each tested rice was counted under the field planting conditions in Beijing and Hainan. The statistical method is: each rice to be tested is planted in 2 rows, randomly arranged, and the heading period of a single plant is counted. The ear of the plant is recorded as heading when 1/2 of the flag leaf sheath is exposed, and the heading period is the number of days from sowing to heading. .

The results showed (Fig. 3, Fig. 4), the heading date of the transgenic OsFTL1 rice was significantly earlier than that of the wild type. Under the long-day conditions in Beijing, the transgenic rice has heading only 45-47 days after sowing, while the wild type is still in the tillering stage. Correspondingly, the heading period of the wild type is 116-118 days after sowing. This shows that under long-day conditions, the heading date of the transgenic rice can be shortened by about 71 days compared with the wild type, and the grain filling maturity period of the rice is correspondingly earlier. Under the short-day conditions in Hainan, the OsFTL1-transgenic rice also headed only 45-50 days after sowing, compared with 58-61 days for the wild type. About 11 days were shortened, and the maturity period of rice grain filling was also advanced accordingly. The results of field experiments in Beijing in 2020 and Hainan in 2021 are shown in Table 1. It shows that OsFTL1 and its coding gene can regulate the heading date of rice.

Table 1. Heading date (days) of wild-type and transgenic rice under field conditions in Beijing and Hainan

rice 2020 Beijing Hainan in 2021 WT 117±1 59.50±1.05 OsFTL1-OE1 46±1** 47.33±0.82** OsFTL1-OE2 45.67±0.58** 46.67±1.21** OsFTL1-OE3 45.33±0.58** 48.67±1.03**

In Table 1, * indicates that compared with WT of the same year, the difference reached a significant level, p < 0.05; ** indicated that compared with WT of the same year, the difference reached a significant level, p < 0.01.

The present invention has been described in detail above. For those skilled in the art, without departing from the spirit and scope of the present invention, and without unnecessary experiments, the present invention can be practiced in a wider range under equivalent parameters, concentrations and conditions. While specific embodiments of the invention have been shown, it should be understood that the invention can be further modified. In a word, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made with conventional techniques known in the art and departing from the disclosed scope of this application. Applications of some of the essential features are possible within the scope of the appended claims below.

sequence listing

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gacatgcgca ccttctacac cctggtgatg gtggatccgg acgcgccgag cccgagcgat 540

ccaaacctca gggagtacct gcactggctg gtcaccgaca tcccggctac cacaggagtc 600

tcttttggga cagaggtggt gtgctacgag agcccgcggc cggtgctggg gatccacagg 660

ctggtgttcc tgctgttcga gcagctgggg cggcagacgg tgtacgcacc ggggtggcgc 720

cagaacttca gcacccgcga cttcgccgag ctctacaacc tcggcctccc tgtcgccgcc 780

gtctacttca actgccagag ggagtctgga accggaggaa gaagaatgtg a 831

<210> 3

<211> 5388

<212>DNA

<213> Rice (Oryza sativa L.)

<400> 3

attcctcacc cctaacttaa agcgagcagc caagctactg tagctaatgg tagggcgcgt 60

gcgacagtgg agtaaagtga agcagcagca gagaagcaga gaccatttcc ctcctccctc 120

ctctctcgcc aaacacgacg acgtcgcctc ggtttagtcg tcgtcggccg gcagccggat 180

gacgggcagt agttgtccgg ccgattcttc ccagctgctg taccctcgcc ggggtgcccc 240

caccaccacc accacctccc gtcctcctct ccatccgctc atcgctcatg cgccctacga 300

cgtcgtcctc caccgatctg tcgtcctctc catcagctca gctcgtgatc aggctgagct 360

cgcgagcttc tggtgctata taaggctggg tggtggtgca agtgcgaagc gagcggccgg 420

tgaggacgat cgatcgagat cgagcttgac ggcggcgaga ggaggaggag gggagacgat 480

gagcgggcgg gggagggggg acccgctggt gctggggagg gtggtggggg acgtggtgga 540

cccgttcgtg aggagggtgg cgctgcgggt ggcgtacgga gcgcgggagg tggccaacgg 600

ctgcgagctc cgcccctccg ccgtcgccga ccagccccgc gtcgccgtcg gcggccccga 660

catgcgcacc ttctacaccc tggtacgtac gtacgcacgc ctccgccgcc atcgccgccg 720

gcgcggcagc tagctgagct gacacactgc tcgatcgatt cttgtggcct ctgcaggtga 780

tggtggatcc ggacgcgccg agcccgagcg atccaaacct cagggagtac ctgcactggt 840

gagtcatcga tcgagtcagc acagcagcag ctgttgtagc tgacgcgccc tgtgacgact 900

gatctgattc agattcagag tccagattca gctccctgca ccgcaccctc gtcgtctcgt 960

gtcgtctcgt ctatcgtgtc gtacgagata gagcgtatag ctatagccag ccaagtgccc 1020

gtgttggtgttgcacttgcc gccacgagat cgatcgagcc ctttttgcct cttttttttt 1080

taaaaaaaaa agagagaata tttttgcatc cgggtagcag ccagtgaata cagttacgaa 1140

acatgaacgt ttgatcgttg actgcatgct agctagctgc ttttcaaaag aaaaagggca 1200

cacatttctt ttaatttaga tgaaaaacaa tatatatgga tcgatcacac attaatctct 1260

ctctgcaaat tctgctatag tgatgtgccg agcactgttt gattgggcct tctagctagt 1320

tttcctcgag ataataacac actcatatgt ataatatact gtatataata tatgatcatg 1380

atctctggaa tgatcattac gtcgtctttg ggagaaaaac acatgatttt acacaaactg 1440

agatatagca aagcaggaac gacattgagt agaacgatct taagatagta tatattcaat 1500

cgatcgtacg taggcaaata aactaattaa ttaatcaact gactgagata ctagctttat 1560

tacagaaaca acaccaaatt tatcagcatt caaacattat tttaatttatt attaatttcc 1620

atggaatgtc tcaaacaatg tcagaaaatc aggtgatgtt ttaagtgcag tatagtagtt 1680

ttttatgtgt acaaaagttc aatatatatt tttcagaatc atttcaataa atacatcata 1740

gctaggatta tcggtaaaat attgtcacat ataattccta gctaggaaaa atatataaaa 1800

caatttgcct gtatctacag tagttgcaca tgctaaggaa ttgtatatat ggattggtag 1860

atattatatt tgtattgcac atccatgaag agataagagg tttaatttta ctcttccatt 1920

atctaaaagg caggtaaaac atttccatta attatctaca aaaagtctaa aaggtaaata 1980

tcatgttgga tatatgtatc aaacgatagg gttttctaaa acattctgtc caacaattta 2040

aaatgtgctt agtttctttttttcccccttg tttagaaaat agtttttatg aagttatttt 2100

gagtggcatt ttccctcaaa tgtggtgtgt atgaaattta ctaaacaatt taacatttga 2160

gcttaaatgc atacacatca catgaaatta atagcaaaac aagttgattt cttctcttcc 2220

tagcttggtt caactaaagt aattaagaga tgataaaata ttatttggat aaaagatggt 2280

gtaaattaaa gtatctatat ctgaaactag agaatagatt ttgatttttc ttgataaatt 2340

tattagagaa gaatttgcag tacacataaa aacaagccaa gaaagtaaaa gtaattaagg 2400

agggaattaa tgtaatgtgc acaattgcta gaggaacatt tgaaaagtaa tccatattta 2460

tttgatgtct tttgcatggt tgcatttccg attaatgttc atatatatgt gaggccttcg 2520

aagtaattta ccaattaaaa tgataatttg catgcacatt ttgtattgta tatagtgatg 2580

acataaaatg attttgattg gcattgctac aaaaatgatt ttggcattgc tgtatatgat 2640

catacggccggggaatattga ttgcaatcag ttgcttttgt caatagttaa ttagaatttt 2700

cgagtgtgaa gggagcttat agacgaactg accgaaacaa taaaattaag aatcctccca 2760

acacaaaacc ctcatgtgca ctgtacgtag atgtgtccaa agatcatgtg caaaatttgt 2820

acaatcaatg aaaataggac cggatgtttc acatatatag ttaattaaat tccaggccat 2880

aattttttaa caaaacaagg ttgattatac taaatataag aaatgattat aagtctattg 2940

ttacaataaa ggacataaaa tttgccaaga caaaccaata tactccatat atagctcaaa 3000

aagtcacaag ctagagatgc gagcagaaga gcgcaaccac aatccttttg agttatttgt 3060

gcaacaaaaa ctctatttct taatatattg acgtgcaatc cttttgcgcg ttcgcgaaaa 3120

agaagagcgc aaccacaaca ttgttgacgt caccaactcc agcacatgca aaagtatata 3180

tgaatgtgta tatgtcacca tcttcgatcc tttgacaaat ctgtaagtca acgtcatcaa 3240

acaaattgga gacccagatt ttaaaatccg caccaccgat tgaatcgtcg gtaacttagc 3300

atacaaatct tcctagatct tcatcgatga tcccagaaat acagaagaca ttgtcaaaga 3360

tatacaaaag acaaaaagaa aaaaaactct agacaatcct aattccatgg acttcctaaa 3420

agacaaatct catctatcta gaatatctaa aattcctttt ttttaagaaa ccctaactaa 3480

aaacttactg gataaagaca agacacgggt ccccttcctc cttgtcaccg gcgaggatac 3540

aggagacgag gagggtacct gatgacggtg gcacatgagg tacccttggc agaggtgagt 3600

catgagcata cgatgctgct atgtgtataa ctgaagaatt tttgttgaga gaattccagg 3660

caataattta cagaaaaccc cattaatgct ggttttaatc cggctaggct agtgccggta 3720

gtgatagtca gtatctctct agttatgagc tagttccaaa ggccatttct atttgttgaa 3780

accctacaat aaaattaata accggttcat cctagctagt aaaaagagaa ttccattata 3840

tatctagta aaatttattc caagacacta atgaaatttg tggccttttt ctctagtaca 3900

ctagttttat ggttatttgt taaaagacac ttgaccgtta gttaatatat attcttccaa 3960

cagagagata aaacttaatt tgcaaaagac aataatcctt gaaccacctt caatataaat 4020

ttgaatattt tttaagctga atttaaactt gtcatacaaa attcaggaat aatatattgt 4080

cccaaattac ggaaatgaga attgccaaaa aaaaatgcaa aatcagaaaa atgaaaaaat 4140

attggatgaa tttaaacgac tttttaaaat gtttttattt ccacaatttt agtgcacatt 4200

ttctacaaag ggaaggagca tataattacg cagattaatc attctacaag tacttagata 4260

aattcttagg aaaactaatt gcccagttct cactatatat attatatgag tagaaaatac 4320

aacgtacact cctagaaata ttttcatgcc aaattaagca aagctagttc ctcattttgc 4380

gaccttcaca tggttagtta ttggttaata tatactccac ttgattccaa ataatgtcag 4440

tcatttcata cttgctcaca agaatcaaga aaacatttca aattacttca ttggaaaatt 4500

aatgtgaacc gatttacgcg aattctttc accctattca aaactgatca acggaacttc 4560

aagttccatg tgctacctat gctgaaaata taatggaact gatcagaatt aacgatctat 4620

caatgcaggc tggtcaccga catcccggct accacaggag tctcttttgg tactgaacaa 4680

ctatagcact ctctgaacaa tcttgcaaca gttacattca tatgtgtgca catcaacaca 4740

tgcatgaaat cagatcactc tgatgtgaag aagagtacaa ttgcttgcta attttgcagg 4800

gacagaggtg gtgtgctacg agagcccgcg gccggtgctg gggatccaca ggctggtgtt 4860

cctgctgttc gagcagctgg ggcggcagac ggtgtacgca ccggggtggc gccagaactt 4920

cagcacccgc gacttcgccg agctctacaa cctcggcctc cctgtcgccg ccgtctactt 4980

caactgccag agggagtctg gaaccggagg aagaagaatg tgagctagct ctcccctctc 5040

tatctctcac tccctgaatt tgggacaagt cctgaatttt gcatgacaaa tcctatactc 5100

ggaacgattt gaaatggtaa aatttgatca tatgtatttt tagctcccta tcctctctct 5160

ctgtacgtta gttacataaa tgctccccacc gattctaaat gtaatttatt tatatttctc 5220

tatttttctc gaaatttaag ttgttatagg acgttgagga actttgtccc ccatttttcc 5280

tttcatacccc ttcattgtta agggaaatgc taccactctg atatatataa aaatgatgta 5340

ccattttcaa tatataataa ccatgcacag ggaatatagt catgttaa 5388

Claims (7)

1. Any one of the following uses of the protein:

d1 Application in regulating and controlling flowering time of rice;

d2 Application of the compound in preparing products for regulating and controlling flowering time of rice;

the protein is a protein with an amino acid sequence of sequence 1.

2. Use of a biological material related to a protein according to claim 1, wherein the biological material is selected from the group consisting of:

d1 Application in regulating and controlling flowering time of rice;

d2 Application of the product in preparing products for regulating and controlling rice flowering time;

the biological material is any one of the following B1) to B7):

b1 A nucleic acid molecule encoding the protein of claim 1;

b2 An expression cassette comprising the nucleic acid molecule according to B1);

b3 A recombinant vector containing the nucleic acid molecule according to B1) or a recombinant vector containing the expression cassette according to B2);

b4 A recombinant microorganism containing the nucleic acid molecule according to B1), or a recombinant microorganism containing the expression cassette according to B2), or a recombinant microorganism containing the recombinant vector according to B3);

b5 A transgenic plant cell line containing the nucleic acid molecule according to B1) or a transgenic plant cell line containing the expression cassette according to B2);

b6 A transgenic plant tissue containing the nucleic acid molecule according to B1) or a transgenic plant tissue containing the expression cassette according to B2);

b7 A transgenic plant organ containing the nucleic acid molecule according to B1) or a transgenic plant organ containing the expression cassette according to B2).

3. Use according to claim 2, characterized in that: b1 The nucleic acid molecule is b 11) or b 12) or b 13) as follows:

b11 A DNA molecule with a coding sequence of a sequence 2 in a sequence table;

b12 A DNA molecule shown as a sequence 2 in a sequence table;

b13 A DNA molecule shown in a sequence 3 in a sequence table.

4. A method for producing a plant with a premature flowering time, comprising expressing the protein of claim 1 in a recipient plant, or increasing the content or activity of the protein of claim 1 in the recipient plant to obtain a plant with a premature flowering time;

the plant rice.

5. The method of claim 4, wherein: the method is carried out by introducing a gene encoding the protein of claim 1 into the recipient plant and expressing the gene.

6. The method of claim 5, wherein: the coding gene is the nucleic acid molecule according to B1) of claim 2 or 3.

7. Use according to any one of claims 1 to 3, or a method according to any one of claims 4 to 6, wherein: the flowering time intermediate is now at the heading stage.

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