Application of a dogtail photoperiod-regulated gene SvEhd1 in shortening the research cycle of rice resistance to blast

Abstract

This invention discloses a photoperiod regulation gene of foxtail grass. SvEhd1 Its application in shortening the research cycle of rice resistance to rice blast falls under the fields of genetic engineering and plant breeding. SvEhd1 The gene nucleotide sequence is shown in SEQ ID NO.1. This invention utilizes cloning... SvEhd1 Genes and their promoters were used to construct recombinant expression vectors, which were then transformed into the blast-susceptible rice variety "Taipei 309," resulting in transgenic rice materials with significantly shortened growth periods. Under natural and artificial long-day conditions, the heading time of the transgenic plants was significantly earlier than that of the wild type, the generation rate was accelerated, and the blast susceptibility of the original variety was not altered. The transgenic materials and methods provided by this invention can effectively shorten the experimental cycle for verifying the function of rice blast resistance genes and evaluating resistance, providing an efficient experimental system for rice disease resistance research.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, specifically to a photoperiod regulation gene in foxtail grass. SvEhd1 Application in shortening the research cycle of rice resistance to rice blast. Background Technology

[0002] Cultivated rice, as one of the world's most important staple crops, provides food for one-third of the global population. However, nearly 30% of rice yield is lost each year due to rice blast. Therefore, elucidating the molecular mechanisms of rice resistance to rice blast and cultivating disease-resistant materials has always been an important research direction in the fields of rice genetics and breeding and plant pathology.

[0003] Currently, research on rice blast resistance mainly relies on techniques such as genetic transformation and gene editing to verify the function and elucidate the mechanisms of candidate resistance genes or regulatory factors. However, rice itself has a long growth cycle, typically requiring three to four months or even longer from sowing to grain filling. Furthermore, the process of obtaining transgenic rice involves multiple steps, including tissue culture, Agrobacterium infection, resistance screening, plant regeneration, and multi-generational stable genetic verification, resulting in a long overall experimental cycle. This significantly restricts the efficiency of functional research on genes related to rice blast resistance. Therefore, the length of the rice growth period is a major limiting factor affecting the time frame of gene function research.

[0004] In existing technologies, research on shortening the rice growth period mainly focuses on using rice population resources or mutant libraries to identify early flowering sites and breed early-maturing varieties. However, the extent of growth period shortening is limited due to genetic diversity within the species. In contrast, there are fewer reports on research and applications of using growth period regulating genes from cross-species sources to modify the rice growth period, especially in the construction of experimental materials for shortening research cycles, where mature and effective technical solutions have not yet been developed.

[0005] Therefore, there is an urgent need to provide a new technical approach that, without altering rice blast resistance, introduces growth period regulating factors into rice varieties that are susceptible to rice blast and have high genetic transformation efficiency, thereby constructing rice experimental materials with significantly shortened growth periods, high transformation efficiency, and suitability for rice blast resistance research, thus accelerating the research process related to rice blast resistance.

[0006] This invention utilizes genetic transformation techniques, combining the conservation and differentiation of genes in the growth period of grasses, to screen out genetic loci that can significantly shorten the growth period of rice. By combining the high genetic transformation efficiency and susceptibility to rice blast of the recipient rice, experimental materials suitable for rapid functional verification of rice blast resistance genes are constructed. Summary of the Invention

[0007] The purpose of this invention is to provide a photoperiod regulation gene for foxtail grass. SvEhd1Application in shortening the research cycle of rice resistance to rice blast.

[0008] To achieve the above-mentioned objectives, the technical solution adopted in this application is as follows: Firstly, this application provides a photoperiod regulation gene from *Setaria viridis*. SvEhd1 Application in the preparation of experimental materials for rice with shortened growth period.

[0009] Secondly, this application provides a photoperiod regulation gene for *Setaria viridis*. SvEhd1 Application in shortening the research cycle of rice resistance to rice blast.

[0010] Thirdly, this application provides a recombinant expression vector for shortening the research cycle of rice resistance to rice blast.

[0011] Fourthly, this application provides a host cell containing a recombinant expression vector.

[0012] Fifthly, this application provides a method for constructing experimental materials for rice blast resistance with a shortened growth period.

[0013] Sixthly, this application provides a transgenic rice experimental material with a shortened growth period, constructed by a method.

[0014] Seventhly, this application provides an application of transgenic rice experimental material in shortening the experimental cycle for verifying the function of rice blast resistance genes or evaluating resistance.

[0015] Eighthly, this application provides a primer set for identifying experimental materials of transgenic rice.

[0016] The first aspect of this application provides a photoperiod regulation gene from *Setaria viridis*. SvEhd1 Application in preparing experimental materials for rice with shortened growth periods; *Setaria viridis* photoperiod regulatory genes. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1. The protein encoded by this gene can significantly shorten the growth period of rice, thereby accelerating the generational succession of rice and shortening the experimental cycle for verifying the function of the rice blast resistance gene. Those skilled in the art can obtain the relevant information through the following methods. SvEhd1 Equivalent genes: (1) obtained through database retrieval; (2) by SvEhd1 Gene fragments were obtained by screening foxtail grass genome libraries or cDNA libraries using probes; (3) according to SvEhd1 Oligonucleotide primers were designed based on gene sequence information, and the genome, mRNA, and cDNA of *Setaria viridis* were obtained by PCR amplification; (4) In SvEhd1 (5) Obtain the gene by modifying it using genetic engineering methods based on the gene sequence; (6) Obtain the gene by chemical synthesis.

[0017] Furthermore, the rice experimental material was Taipei 309, a rice variety susceptible to rice blast.

[0018] The second aspect of this application provides a photoperiod regulation gene from *Setaria viridis*. SvEhd1 Application in shortening the research cycle of rice blast resistance, the photoperiod regulatory gene of foxtail grass. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1.

[0019] Furthermore, applications include the use of foxtail grass photoperiod regulation genes. SvEhd1 Transgenic rice materials with significantly shortened growth periods were obtained by transferring the genetic material into rice, which can be used for verification of the function of rice blast resistance genes or for resistance evaluation.

[0020] A third aspect of this application provides a recombinant expression vector for shortening the research cycle of rice resistance to rice blast, the recombinant expression vector containing a photoperiod regulation gene from *Setaria viridis*. SvEhd1 and its promoter, including the foxtail grass photoperiod regulatory gene. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1, and the nucleotide sequence of the promoter is shown in SEQ ID NO.3. The isolated and cloned... SvEhd1 The gene encodes a type B response regulatory protein, comprising a phosphoacceptor receiver (REC) domain and a MYB-like DNA-binding domain. This invention constructs a [protein name missing] using molecular cloning technology. SvEhd1 The recombinant expression vector of the gene was used to obtain transgenic positive homozygous lines in the rice variety "Taipei 309". SvEhd1 The genetically modified homozygous lines had a significantly shorter growth period than the wild-type "Taipei 309" plant under both natural and artificial long-day conditions in Nanjing, indicating that... SvEhd1 This shortened the growth period of rice and verified that... SvEhd1 Its role in constructing research materials for extremely early-maturing rice blast resistance. Given... SvEhd1 The role of this gene in regulating rice growth period suggests that it has potential application value in breeding extremely early-maturing, blast-resistant rice experimental materials. This invention also provides a basis for utilizing… SvEhd1 This laid the theoretical and applied foundation for the cultivation of extremely early maturing varieties.

[0021] Furthermore, the vector was the plant expression vector pCAMBIA1301.

[0022] The fourth aspect of this application provides a host cell containing a recombinant expression vector, wherein the host cell is Agrobacterium tumefaciens EHA105.

[0023] The fifth aspect of this application provides a method for constructing experimental materials for rice blast resistance with a shortened growth period, comprising the following steps: (1) Cloning the photoperiod regulatory gene of foxtail grass as shown in SEQ ID NO.1 SvEhd1 and its promoter as shown in SEQ ID NO.3; (2) The gene and promoter are constructed into a plant expression vector to obtain a recombinant expression vector; (3) The recombinant expression vector was transformed into Taipei 309, a rice variety susceptible to rice blast, to obtain transgenic rice plants; (4) Screening to obtain homozygous transgenic rice materials with significantly shortened growth period.

[0024] Furthermore, in step (3), the transformation method is Agrobacterium-mediated genetic transformation.

[0025] The sixth aspect of this application provides a transgenic rice experimental material with a shortened growth period, constructed by a method, wherein the material is transgenic with the rice shown in SEQ ID NO.1. SvEhd1 Homozygous positive plants of Taipei 309 rice.

[0026] Furthermore, the growth period of the transgenic rice experimental material was significantly shorter than that of the wild-type Taipei 309 under natural long-day or artificial long-day conditions.

[0027] The seventh aspect of this application provides the application of transgenic rice experimental materials in shortening the experimental cycle for verifying the function of rice blast resistance genes or evaluating resistance.

[0028] The eighth aspect of this application provides a primer set for identifying experimental materials of transgenic rice. The primer set includes an upstream primer and a downstream primer. The nucleotide sequence of the upstream primer is shown in SEQ ID NO.4, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.5.

[0029] Beneficial effects: This invention utilizes the photoperiod regulation gene of foxtail grass. SvEhd1 Transgenic rice materials were obtained by introducing the blast-susceptible rice variety "Taipei 309". This significantly shortens the rice growth period, accelerates generational turnover, and does not alter the recipient variety's blast susceptibility. It effectively shortens the research cycle for rice blast resistance, provides a standardized experimental material system for early-maturing rice, and offers a highly versatile and easily applicable method, providing genetic resources and a material foundation for the breeding of early-maturing rice varieties.

[0030] Compared with the prior art, the present invention has the following advantages: (1) The present invention regulates the photoperiod of foxtail grass by... SvEhd1Transgenic rice materials were obtained by introducing the blast-susceptible rice variety "Taipei 309". Experimental results showed that under natural long-day conditions in Nanjing, the average heading time of the transgenic plants was 46.7 days, nearly half that of the wild-type "Taipei 309" (95.6 days); under artificial long-day conditions, the average heading time of the transgenic plants was 74.8 days, significantly shorter than that of the wild-type (255.7 days). This indicates... SvEhd1 Genes can significantly shorten the rice growth period and accelerate the generational turnover rate.

[0031] (2) The recipient variety selected in this invention, “Taipei 309”, is a rice blast-sensitive variety with high genetic transformation efficiency. While significantly shortening the growth period, the transgenic material retains the rice blast-sensitive characteristics of the original variety, making it suitable for functional verification of rice blast-related genes and resistance evaluation studies, thus avoiding interference factors introduced by changing the disease resistance background.

[0032] (3) Research on rice resistance to rice blast usually involves gene function verification and multi-generation genetic stability analysis. Due to the long growth cycle of rice, the overall experimental cycle is long and the efficiency is low. The transgenic rice material provided by this invention has a significantly shortened growth period, and can complete the entire life cycle from sowing to grain filling in a shorter time. This significantly accelerates the experimental process of functional verification, genetic analysis, and resistance evaluation of genes related to rice blast resistance, and improves research efficiency.

[0033] (4) The invention constructs SvEhd1 The transgenic "Taipei 309" material possesses characteristics such as a clear genetic background, a stable and shortened growth period, and maintained susceptibility to rice blast disease. It can be used as a standardized experimental material for rice blast resistance research. This material facilitates comparison and verification between different laboratories, contributing to the standardization and efficiency of rice disease resistance research.

[0034] (5) The transgenic material construction method provided by this invention is not limited to a specific promoter type or transformation method, and has strong versatility and scalability. In addition to "Taipei 309", this method can also be applied to other rice varieties with high genetic transformation efficiency and susceptibility to rice blast, providing a technical path for constructing early-maturing rice materials suitable for different research purposes.

[0035] (6) This invention verifies the source of foxtail grass. SvEhd1 The gene has the function of significantly shortening the growth period in rice, providing a new cross-species gene resource for early-maturing rice breeding. At the same time, the obtained transgenic material can also be used as intermediate breeding material to cultivate new early-maturing rice varieties. Attached Figure Description

[0036] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0037] Figure 1 For the present invention SvEhd1 Gene structure diagram; Figure 2 For the carrying of the present invention SvEhd1 Recombinant vector structure diagram of gene and its promoter sequence; Figure 3 Wild-type "Taipei 309" (TP309) provided for embodiments of the present invention and SvEhd1 Transgenic plant (TP309:: SvEhd1 (A statistical chart showing the reproductive period under natural and artificial long-day conditions in Nanjing during summer.) Figure 4 Wild-type "Taipei 309" (TP309) provided for embodiments of the present invention and SvEhd1 Transgenic plant (TP309:: SvEhd1 Phenotypic diagram under natural long daylight in Nanjing during summer. Detailed Implementation

[0038] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar to or equivalent to those described herein may be used in the implementation or testing of this invention. Reagents not specifically described in detail herein are conventional reagents and are commercially available; methods not specifically described in detail are conventional experimental methods and are known from the prior art.

[0039] The first aspect of this application provides a photoperiod regulation gene from *Setaria viridis*. SvEhd1 Application in preparing experimental materials for rice with shortened growth periods; *Setaria viridis* photoperiod regulatory genes. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1. The protein encoded by this gene can significantly shorten the growth period of rice, thereby accelerating the generational succession of rice and shortening the experimental cycle for verifying the function of the rice blast resistance gene. Those skilled in the art can obtain the relevant information through the following methods. SvEhd1 Equivalent genes: (1) obtained through database retrieval; (2) by SvEhd1 Gene fragments were obtained by screening foxtail grass genome libraries or cDNA libraries using probes; (3) according to SvEhd1Oligonucleotide primers were designed based on gene sequence information, and the genome, mRNA, and cDNA of *Setaria viridis* were obtained by PCR amplification; (4) In SvEhd1 (5) Obtain the gene by modifying it using genetic engineering methods based on the gene sequence; (6) Obtain the gene by chemical synthesis.

[0040] In some embodiments, the rice experimental material is Taipei 309, a rice variety susceptible to rice blast.

[0041] A second aspect of this application provides a photoperiod regulation gene from *Setaria viridis*. SvEhd1 Application in shortening the research cycle of rice blast resistance, the photoperiod regulatory gene of foxtail grass. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1.

[0042] In some embodiments, the application includes using the photoperiod regulation gene of foxtail grass. SvEhd1 Transgenic rice materials with significantly shortened growth periods were obtained by transferring the genetic material into rice, which can be used for verification of the function of rice blast resistance genes or for resistance evaluation.

[0043] A third aspect of this application provides a recombinant expression vector for shortening the research cycle of rice blast resistance, the recombinant expression vector containing a foxtail grass photoperiod regulatory gene. SvEhd1 and its promoter, including the foxtail grass photoperiod regulatory gene. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1, and the nucleotide sequence of the promoter is shown in SEQ ID NO.3. The isolated and cloned... SvEhd1 The gene encodes a type B response regulatory protein, which includes a phosphoacceptor receiver (REC) domain and a MYB-like DNA binding domain.

[0044] In some embodiments, the vector is the plant expression vector pCAMBIA1301.

[0045] The fourth aspect of this application provides a host cell containing a recombinant expression vector, wherein the host cell is Agrobacterium tumefaciens EHA105.

[0046] The fifth aspect of this application provides a method for constructing experimental materials for rice with shortened growth period and resistance to rice blast, comprising the following steps: (1) Cloning the photoperiod regulatory gene of foxtail grass as shown in SEQ ID NO.1 SvEhd1 and its promoter as shown in SEQ ID NO.3; (2) The gene and promoter are constructed into a plant expression vector to obtain a recombinant expression vector; (3) The recombinant expression vector was transformed into Taipei 309, a rice variety susceptible to rice blast, to obtain transgenic rice plants; (4) Screening to obtain homozygous transgenic rice materials with significantly shortened growth period.

[0047] In some embodiments, in step (3), the transformation method is Agrobacterium-mediated genetic transformation.

[0048] The sixth aspect of this application provides a transgenic rice experimental material with a shortened growth period, constructed by a method, wherein the material is transgenic with the rice shown in SEQ ID NO.1. SvEhd1 Homozygous positive plants of Taipei 309 rice.

[0049] In some embodiments, the growth period of transgenic rice experimental materials under natural or artificial long-day conditions is significantly shorter than that of wild-type Taipei 309.

[0050] The seventh aspect of this application provides the application of transgenic rice experimental materials in shortening the experimental cycle for verifying the function of rice blast resistance genes or evaluating resistance.

[0051] The eighth aspect of this application provides a primer set for identifying experimental materials of transgenic rice. The primer set includes an upstream primer and a downstream primer. The nucleotide sequence of the upstream primer is shown in SEQ ID NO.4, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.5. Example 1

[0052] A photoperiod regulation gene of foxtail grass according to the present invention SvEhd1 Application in preparing experimental materials for rice with shortened growth periods; *Setaria viridis* photoperiod regulatory genes. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1. The protein encoded by this gene can significantly shorten the growth period of rice, thereby accelerating the generational succession of rice and shortening the experimental cycle for verifying the function of the rice blast resistance gene. Those skilled in the art can obtain the relevant information through the following methods. SvEhd1 Equivalent genes: (1) obtained through database retrieval; (2) by SvEhd1 Gene fragments were obtained by screening foxtail grass genome libraries or cDNA libraries using probes; (3) according to SvEhd1 Oligonucleotide primers were designed based on gene sequence information, and the genome, mRNA, and cDNA of *Setaria viridis* were obtained by PCR amplification; (4) In SvEhd1 (5) Obtain the gene by modifying it using genetic engineering methods based on the gene sequence; (6) Obtain the gene by chemical synthesis.

[0053] The rice experimental material was Taipei 309, a rice variety susceptible to rice blast.

[0054] A photoperiod regulation gene of foxtail grass according to the present invention SvEhd1 Application in shortening the research cycle of rice blast resistance, the photoperiod regulatory gene of foxtail grass. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1.

[0055] Applications include the use of foxtail grass photoperiod regulation genes. SvEhd1 Transgenic rice materials with significantly shortened growth periods were obtained by transferring the genetic material into rice, which can be used for verification of the function of rice blast resistance genes or for resistance evaluation.

[0056] This invention provides a recombinant expression vector for shortening the research cycle of rice blast resistance, the recombinant expression vector containing a foxtail grass photoperiod regulatory gene. SvEhd1 and its promoter, including the foxtail grass photoperiod regulatory gene. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1, and the nucleotide sequence of the promoter is shown in SEQ ID NO.3. The isolated and cloned... SvEhd1 The gene encodes a type B response regulatory protein, which includes a phosphoacceptor receiver (REC) domain and a MYB-like DNA binding domain.

[0057] The vector was the plant expression vector pCAMBIA1301.

[0058] The present invention provides a host cell containing a recombinant expression vector, wherein the host cell is Agrobacterium tumefaciens EHA105.

[0059] This invention provides SvEhd1 Application of transgenic experimental materials in shortening the research cycle of rice blast disease; in the specific implementation of this invention, the... SvEhd1 The gene is located on chromosome 9 of *Setaria viridis* and contains a phosphoacceptor receiver (REC) domain and a MYB-like DNA-binding domain. The gene structure is as follows: Figure 1 As shown; the SvEhd1 The nucleotide sequence of the gene is shown in SEQ ID NO.1, specifically: In the specific implementation of this invention, the SvEhd1 The amino acid sequence encoded by the gene is shown in SEQ ID NO.2, specifically: In the specific implementation of this invention, the SvEhd1 The promoter sequence of the gene is shown in SEQ ID NO.3, specifically: The present invention also provides a method for identifying the above-described scheme. SvEhd1The primer set for the genetically modified plant is characterized by comprising an upstream primer with nucleotide sequences as shown in SEQ ID NO.4 and a downstream primer as shown in SEQ ID NO.5.

[0060] The primer sequences for specific identification of the vector insert fragment are as follows: Upstream primer: 5'-CAAGCGATCATACATATCTAAG-3' (SEQ ID NO.4); Downstream primer: 5'-CATGAATCCCAACTGTAGC-3' (SEQ ID NO.5).

[0061] To further illustrate the present invention, the following description, in conjunction with the accompanying drawings and embodiments, explains the invention. SvEhd1 The application of genes in the development of experimental materials for the breeding of very early-maturing rice resistant to rice blast is described in detail, but it should not be construed as limiting the scope of protection of this invention. Example 2

[0062] The present invention provides a method for constructing experimental materials for rice with shortened growth period and resistance to rice blast, comprising the following steps: (1) Cloning the photoperiod regulatory gene of foxtail grass as shown in SEQ ID NO.1 SvEhd1 and its promoter as shown in SEQ ID NO.3; (2) Genes and promoters are constructed into plant expression vectors to obtain recombinant expression vectors; (3) The recombinant expression vector was transformed into Taipei 309, a rice variety susceptible to rice blast, to obtain transgenic rice plants; the transformation method was Agrobacterium-mediated genetic transformation.

[0063] (4) Screening to obtain homozygous transgenic rice materials with significantly shortened growth period.

[0064] This invention discloses a transgenic rice experimental material with a shortened growth period, constructed by a method thereof. The material is transgenic with the rice shown in SEQ ID NO.1. SvEhd1 Homozygous positive plants of Taipei 309 rice.

[0065] The growth period of the transgenic rice experimental materials was significantly shorter than that of the wild-type Taipei 309 under natural or artificial long-day conditions.

[0066] The present invention relates to the application of a transgenic rice experimental material in shortening the experimental cycle for verifying the function of rice blast resistance genes or evaluating resistance.

[0067] The present invention provides a primer set for identifying experimental materials of transgenic rice. The primer set includes an upstream primer and a downstream primer. The nucleotide sequence of the upstream primer is shown in SEQ ID NO.4, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.5. Experimental Example 1

[0068] SvEhd1 Gene cloning, vector construction, and identification of positive plants 1. Photoperiod-related genes in foxtail grass SvEhd1 Isolation and cloning: to obtain SvEhd1 The gene contains the full-length sequence of the promoter. Using the *Setaria viridis* genome sequence from a public database as a reference sequence, primers were designed (all primers have a BamHI restriction site at the 5' end). The forward primer sequence is shown in SEQ ID NO. 6; the reverse primer sequence is shown in SEQ ID NO. 7. Using the *Setaria viridis* cultivar “ME034v” as a template, the candidate gene fragment was amplified using PCR technology. The PCR program was as follows: 95℃ pre-denaturation for 5 minutes, 95℃ denaturation for 30 seconds, 60℃ annealing for 45 seconds, 68℃ extension for 10 minutes, for a total of 33 cycles, followed by incubation at 72℃ for 10 minutes, and finally isothermal treatment at 10℃. The PCR products were then recovered by gel extraction.

[0069] The amplification primer sequences are as follows: Upstream primer: 5'-AGCTCGGTACCCGGGGATCCCTAACGGCAACGGAACAT-3' (SEQ ID NO.6); Downstream primer: 5'-AGGTCGACTCTAGAGGATCCCGCCCTGACATAATCTGAAT-3' (SEQ ID NO. 7); 2. SvEhd1 Ligation of the gene to the basic vector: The PCR product and the basic vector pCAMBIA1301 plasmid were simultaneously digested with the restriction endonuclease BamHI and purified. The candidate gene fragment was then ligated into the basic vector pCAMBIA1301 using homologous recombinase.

[0070] 3. PCR molecular detection: Using the transformed DNA as a template, PCR reaction was performed using primers specific to the vector insert fragment. The primer sequences for the insert fragment are shown in the sequence listing; the forward primer sequence is shown in SEQ ID NO.4; and the reverse primer sequence is shown in SEQ ID NO.5. Experimental Example 2

[0071] verify SvEhd1 The role of genes in the growth period of rice In order to determine SvEhd1The regulatory role of genes on the growth period of rice, using wild-type "Taipei 309" and transgenic material TP309:: SvEhd1 As experimental materials, they were planted in Nanjing summer fields under natural long-day and artificial long-day (16 hours of light, 8 hours of darkness) conditions to observe the length of their respective growth periods.

[0072] Under the natural long-day conditions in Nanjing, such as Figure 3 As shown, the average heading time of the wild-type "Taipei 309" is 95.6 days, while that of TP309:: SvEhd1 The average heading time was 46.7 days, nearly half that of the wild type. The phenotypic differences between the two are as follows: Figure 4 As shown, TP309:: SvEhd1 The earlier heading time results in shorter plants compared to the wild type "Taipei 309". Under artificial long-day conditions, such as... Figure 3 As shown, the average heading time of the wild-type "Taipei 309" is 255.7 days, while that of TP309:: SvEhd1 The average heading time was 74.8 days, significantly shorter than the wild-type growth period. Based on these results, it can be concluded that the transgenic line TP309:: SvEhd1 It has a shorter growth cycle than the wild type. SvEhd1 It has an early-maturing effect on rice.

[0073] In summary, this invention utilizes genetic engineering technology to target genes related to photoperiod in foxtail grass. SvEhd1 By introducing the universally susceptible, high-genetic-transformation-rate rice variety "Taipei 309," experiments have verified that, under both natural and artificial long-day conditions in Nanjing, transgenic plants exhibit earlier heading and a shorter growth cycle compared to wild-type plants. This invention is the first experimental confirmation of this... SvEhd1 This gene can significantly shorten the rice growth period. Applying it to the modification of rice varieties with high susceptibility and high conversion rates helps shorten the experimental cycle for verifying functional genes for rice blast resistance and assessing resistance. Simultaneously, this invention also provides a means to utilize… SvEhd1 Genetically bred early-maturing rice varieties have laid a solid theoretical and applied foundation.

[0074] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope. The scope of protection of the present invention is defined by the appended claims, specification, and their equivalents.

Claims

1. A photoperiod regulatory gene in foxtail grass SvEhd1 Its application in preparing experimental materials for rice with shortened growth period is characterized by: The photoperiod regulatory gene of foxtail grass SvEhd1 The nucleotide sequence is shown in SEQ ID NO.

1.

2. The application according to claim 1, characterized in that: The rice experimental material was Taipei 309, a rice variety susceptible to rice blast.

3. A photoperiod regulatory gene in foxtail grass SvEhd1 Its application in shortening the research cycle of rice resistance to rice blast is characterized by: The photoperiod regulatory gene of foxtail grass SvEhd1 The nucleotide sequence is shown in SEQ ID NO.

1.

4. The application according to claim 3, characterized in that: The application includes the use of foxtail grass photoperiod regulation genes. SvEhd1 Transgenic rice was introduced into rice plants to obtain transgenic rice materials with significantly shortened growth periods.

5. A recombinant expression vector for shortening the research cycle of rice resistance to rice blast, characterized in that: The recombinant expression vector contains a photoperiod regulation gene from *Setaria viridis*. SvEhd1 and its promoter, including the foxtail grass photoperiod regulatory gene. SvEhd1 The nucleotide sequence is shown in SEQ ID NO.1, and the nucleotide sequence of the promoter is shown in SEQ ID NO.

3.

6. The recombinant expression vector according to claim 5, characterized in that: The vector is the plant expression vector pCAMBIA1301.

7. A host cell containing the recombinant expression vector of claim 5 or 6, characterized in that: The host cell was Agrobacterium tumefaciens EHA105.

8. A method for constructing experimental materials for rice blast resistance with a shortened growth period, characterized in that... Includes the following steps: (1) Cloning the photoperiod regulatory gene of foxtail grass as shown in SEQ ID NO.1 SvEhd1 and its promoter as shown in SEQ ID NO. 3; (2) The gene and promoter are constructed into a plant expression vector to obtain a recombinant expression vector; (3) The recombinant expression vector was transformed into Taipei 309, a rice variety susceptible to rice blast, to obtain transgenic rice plants; (4) Screening to obtain homozygous transgenic rice materials with significantly shortened growth period.

9. The method according to claim 8, characterized in that: In step (3), the transformation method is Agrobacterium-mediated genetic transformation.

10. A transgenic rice experimental material with a shortened growth period, constructed by the method of claim 8, characterized in that: The material described is transferred as shown in SEQ ID NO.

1. SvEhd1 Homozygous positive plants of Taipei 309 rice.

11. The transgenic rice experimental material according to claim 10, characterized in that: The genetically modified rice experimental material had a significantly shorter growth period than the wild-type Taipei 309 under natural or artificial long-day conditions.

12. The application of the transgenic rice experimental material according to claim 10 or 11 in shortening the experimental cycle for verifying the function of rice blast resistance genes or evaluating resistance.

13. A primer set for identifying the transgenic rice experimental material of claim 12, characterized in that: The primer set includes an upstream primer and a downstream primer. The nucleotide sequence of the upstream primer is shown in SEQ ID NO.4, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.5.

Images