Application of RTMF1 as anti-temperature-sensitive male sterility gene in rice production

Abstract

The invention relates to the technical field of rice production, and discloses application of RTMF1 as a rice anti-thermo-sensitive male sterility gene in rice production, the sequence of the RTMF1 gene is as shown in SEQ ID NO.1, and the coded amino acid sequence is as shown in SEQ ID NO.2. The mutant rtmf1 carrying RTMF1 gene mutation is sterile at a low temperature and a normal temperature, is fertile at a high temperature, has an anti-thermo-sensitive characteristic, is opposite to a mainstream thermo-sensitive sterile line in fertility conversion mode and has a complementary characteristic, seed production is performed at the low temperature or the normal temperature by utilizing the anti-thermo-sensitive characteristic of the RTMF1 gene mutant, the problem of fertility recovery caused by the low temperature can be avoided, and the production efficiency is improved. The seed production risk is reduced and the seed production purity is improved.

Description

Technical Field

[0001] This invention relates to the field of rice production technology, and more specifically, to... RTMF1 Application of temperature-sensitive male sterility genes in rice production. Background Technology

[0002] Rice (Oryza sativa L.), as one of the major food crops, provides essential nutrients for human survival, and its high and stable yields are a crucial guarantee for global food security. Heterosis allows hybrid rice varieties to yield higher yields than self-pollinated varieties, but the hermaphroditism of rice limits the application of heterosis. In the 1970s, Yuan Longping's team published a paper on male sterility in rice and pioneered the discovery of "wild abortive" male sterile cytoplasm. Since then, heterosis has been effectively utilized in rice production, and hybrid rice yields have significantly increased. The application of male sterility in hybrid rice has continued to develop, from the "three-line method" based on cytoplasmic male sterility to the "two-line method" mainly utilizing environmentally sensitive male sterility (EGMS), further overcoming the limitations of the "three-line method" due to the constraints of restoration and conservation relationships and the cumbersome breeding procedures.

[0003] The two-line hybrid rice method, utilizing EGMS, has significant advantages in hybridization breeding, with its planting area now exceeding half. EGMS has rapidly developed and is widely used in hybrid rice, with photoperiod-temperature-sensitive male sterility (P / TGMS) being a commonly used type. P / TGMS generally refers to male sterility under long-day / high-temperature conditions and fertility under short-day / low-temperature conditions. Currently, TGMS is the primary EGMS used in large-scale seed production, and most two-line hybrid rice varieties utilize photoperiod-sensitive male sterility genes. TMS5 However, in recent years, extreme weather events have become more frequent, leading to fluctuations in fertility in two-line male-sterile lines based on TGMS, increasing seed production risks, especially in the rice-growing areas of the upper Yangtze River in my country. Reverse thermosensitive genic male-sterile lines (RTMS), which are sterile at low temperatures (generally ≤28℃) and recover fertility at high temperatures (generally ≥30℃), can compensate for the shortcomings of positive thermosensitive lines and complement TGMS, showing clear application potential. Therefore, exploring RTMS gene resources is extremely important for two-line breeding and can provide a new approach to utilizing heterosis in rice. However, very few RTMS genes or loci have been reported so far. J207S exhibits complete male sterility below 31℃, however... RTMS1 It has not yet been cloned, but has only been located on chromosome 10, in YnS. RTMS10It also exhibits anti-thermosensitive characteristics and is located on chromosome 10. Currently, only one gene with anti-thermosensitive characteristics has been reported: ARGONAUTE 1d (OsAGO1d), whose expression is induced by low temperature. Its encoded protein can bind miR2118 and miR2275 and promote phasiRNA synthesis. At low temperatures, its function is lost, leading to insufficient phasiRNA. Thermosensitive TMS two-line male-sterile lines are easily affected by the environment, resulting in unstable fertility and increased seed production risks. Anti-thermosensitive RTMS gene resources are scarce, and the mechanism is unclear, limiting further application. Because the fertility of two-line male-sterile lines is easily affected by environmental temperature, low temperatures during the rice heading stage may lead to fertility recovery, causing seed production failure and posing a significant production risk to seed companies. RTMS and TGMS show opposite trends: fertile at high temperatures and sterile at low temperatures, complementing thermosensitive characteristics. However, very few sterile materials have been discovered so far, resulting in a scarcity of resources.

[0004] Based on this, we provide RTMF1 The application of temperature-sensitive male sterility genes in rice production has important practical significance. Summary of the Invention

[0005] In view of this, the present invention proposes RTMF1 The application of temperature-sensitive male sterility genes in rice production aims to solve at least one of the aforementioned background technical problems.

[0006] This invention provides a rice temperature-sensitive male sterility gene RTMF1, the sequence of which is shown in SEQ ID NO.1, and the encoded amino acid sequence is shown in SEQ ID NO.2. The mutant gene of RTMF1 is... rtmf1, That The sequence is shown in SEQ ID NO.3, and the encoded amino acid sequence is shown in SEQ ID NO.4.

[0007] The present invention also provides an application of the rice temperature-sensitive male sterility gene RTMF1 and its mutant gene in rice production, including using rice sterile lines carrying RTMF1 gene mutations for seed production and propagation, or introducing them into different rice genetic backgrounds through hybridization and backcrossing to cultivate new temperature-sensitive male sterile lines.

[0008] The present invention also provides a method for cultivating a temperature-sensitive male-sterile rice line, which includes mutating the RTMF1 gene in a normal rice variety.

[0009] Preferably, the RTMF1 gene of rice varieties is mutated using physical, chemical, or biological techniques. The mutations include various types of mutations that cause premature termination of translation of the protein encoded by the gene, changes in amino acids, or changes in gene expression levels.

[0010] This invention also provides the application of the temperature-sensitive male-sterile rice line described above in rice hybrid seed production, self-pollination propagation, and hybridization to cultivate new male-sterile lines.

[0011] The present invention also provides biomaterials related to the rice temperature-sensitive male sterility gene RTMF1 described in the above technical solution, characterized in that the biomaterials are any one of the following (1) to (4): (1) An expression cassette containing the rice temperature-sensitive male sterility gene RTMF1 or a partial continuous sequence as described in claim 1; (2) A recombinant vector containing the rice temperature-sensitive male sterility gene RTMF1 or a partial continuous sequence as described in claim 1; (3) Recombinant microorganisms containing the rice thermosensitive male sterility gene RTMF1 or a partial continuous sequence as described in claim 1; (4) Recombinant microorganisms containing the recombinant vector described in (2).

[0012] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) The present invention carries RTMS The mutant strain with the gene mutation is infertile at low and normal temperatures, but fertile at high temperatures, exhibiting anti-thermosensitive characteristics. This is the opposite of the fertility conversion mechanism of mainstream thermosensitive male-sterile lines, and it also possesses complementary characteristics. RTMF1 The temperature-sensitive properties of gene mutants allow for seed production at low or normal temperatures, avoiding fertility recovery issues caused by low temperatures, reducing seed production risks, and improving seed purity.

[0013] (2) By transferring this gene in different backgrounds, sterile lines with suitable fertility conversion temperatures can be obtained, expanding its application range in different ecological regions and providing a new and effective way to utilize the heterosis of rice. Attached Figure Description

[0014] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings: Figure 1 WT at room temperature in Example 1 and rtmf1 A schematic diagram of fertility-related phenotypic results; Figure 2 WT at high and low temperatures rtmf1 A schematic diagram of fertility-related phenotypic results; Figure 3 In Example 2 RTMF1 A schematic diagram of the gene cloning and functional verification results; Figure 4 In Example 3 rtmf1 A schematic diagram of genetic analysis results in other contexts. Detailed Implementation

[0015] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.

[0016] Furthermore, regarding the numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included within this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0017] 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 or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0018] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This specification and embodiments are merely exemplary.

[0019] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0020] This invention proposes RTMF1 As an application of the temperature-sensitive male sterility gene in rice production, the aforementioned RTMF1 The gene sequence is shown in SEQ ID NO.1, and the encoded amino acid sequence is shown in SEQ ID NO.2. RTMF1 The mutated gene is rtmf1, Its sequence is shown in SEQ ID NO.3, and the encoded amino acid sequence is shown in SEQ ID NO.4.

[0021] The specific SEQ ID NO.1 is: ATGACGGCGCCGGCGGACAAGGGGAAGAAGGCCAAGACCGACGCCGACGGCGGCGCCGCCGAGGAGAACGAGCAGATCGACGGCGCCCTCGTCCTCTCCATCGAGAAGCTCCAGGAGATCCAGGACGAGCTCGAGAAGGTCAATGAGGAAGCTAGTGACAAGGTTTTGGAGGTCGAGCAGAAATACAGTGAGATTCGCAGACCTGTCTATCTCCGAAGGAGTGACGTTATCCAAACAATCCCCGACTTCTGGCTGACAGCGTTTCTGAGTCATCCTCTACTTAGTGAGCTTTTGACCGAAGAGGATCAAAAGATGTTCAAGTACCTGGAGTCTGTCGACGTGGATGATTCTAAAGATGTCAAGTCAGGCTACTCCATAACTCTTACCTTCTCCGAGAACCCGTACTTTGAAGACAAAGAGCTCACGAAGACATATGCCTTCGCTGATGACGGAACAACCACAATAAATGCTACTAGCATTAAGTGGAAAGAAGGAATGGAAATTGCAAATGGGAATGCCAAGAAGAAAGGGAGCAAGCGACCATTGGTTGAGGAAAGTTTCTTCACCTGGTTTACTGATACAGAGCACAAGAGTCTTGCTGATGGTGTGCAAGATGAGGTGGCTGAGATCATCAAGGAAGACCTGTGGCCCAATCCATTGAAGTATTTCAATAATGAGGCTGAAGAGTTAGGAGAGGATGACGACGAAGAGGGGTCTGATGCTGATGAGGGTGAAGAGGATGAGGAGGAGGAGAACTGA; The specific SEQ ID NO.2 is as follows: MTAPADKGKKAKTDADGGAAEENEQIDGALVLSIEKLQEIQDELEKVNEEASDKVLEVEQKYSEIRRPVYLRRSDVIQTIPDFWLTAFLSHPLLSELLTEEDQKMFKYLESVDVDDSKDVKSGYSITLTFSENPYFEDKELTKTYAFADDGTTTINATSIKWKEGMEIANGNAKKKGSKRPLVEESFFTWFTDTEHKSLADGVQDEVAEIIKEDLWPNPLKYFNNEAEELGEDDDEEGSDADEGEEDEEEEN。

[0022] SEQ ID NO.3 is specifically: ATGACGGCGCCGGCGGACAAGGGGAAGAAGGCCAAGACCGACGCCGACGGCGGCGCCGCCGAGGAGAACGAGCAGATCGACGGCGCCCTCGTCCTCTCCATCGAGAAGCTCCAGGAGATCCAGGACGAGCTCGAGAAGGTCAATGAGGAAGCTAGTGACAAGGTTTTGGAGGTCGAGCAGAAATACAGTGAGATTCGCAGACCTGTCTATCTCTGAAGGAGTGACGTTATCCAAACAATCCCCGACTTCTGGCTGACAGCGTTTCTGAGTCATCCTCTACTTAGTGAGCTTTTGACCGAAGAGGATCAAAAGATGTTCAAGTACCTGGAGTCTGTCGACGTGGATGATTCTAAAGATGTCAAGTCAGGCTACTCCATAACTCTTACCTTCTCCGAGAACCCGTACTTTGAAGACAAAGAGCTCACGAAGACATATGCCTTCGCTGATGACGGAACAACCACAATAAATGCTACTAGCATTAAGTGGAAAGAAGGAATGGAAATTGCAAATGGGAATGCCAAGAAGAAAGGGAGCAAGCGACCATTGGTTGAGGAAAGTTTCTTCACCTGGTTTACTGATACAGAGCACAAGAGTCTTGCTGATGGTGTGCAAGATGAGGTGGCTGAGATCATCAAGGAAGACCTGTGGCCCAATCCATTGAAGTATTTCAATAATGAGGCTGAAGAGTTAGGAGAGGATGACGACGAAGAGGGGTCTGATGCTGATGAGGGTGAAGAGGATGAGGAGGAGGAGAACTGA。

[0023] SEQ ID NO.4 is specifically: MTAPADKGKKAKTDADGGAAEENEQIDGALVLSIEKLQEIQDELEKVNEEASDKVLEVEQKYSEIRRPVYL。

[0024] The present invention also provides an application of the rice temperature-sensitive male sterility gene RTMF1 and its mutant gene in rice production, including using rice sterile lines carrying RTMF1 gene mutations for seed production and propagation, or introducing them into different rice genetic backgrounds through hybridization and backcrossing to cultivate new temperature-sensitive male sterile lines.

[0025] In this invention, the seed production is preferably carried out under conditions of low temperature ≤28℃ or normal temperature ≤28℃.

[0026] In this invention, the propagation is preferably carried out under high temperature conditions of ≥30°C.

[0027] This invention also provides a method for cultivating a temperature-sensitive male-sterile rice line, comprising: RTMF1 The gene mutation was introduced into rice varieties, RTMF1 The gene sequences are shown in SEQ ID NO.1 and SEQ ID NO.2.

[0028] In this invention, the RTMF1 Gene mutations include those induced in rice varieties using physical (such as various types of radiation), chemical (such as EMS mutagenesis), or biological techniques (such as gene editing). RTMF1 Genes can mutate, and mutations can take many forms, including those that cause premature termination of the translation of the protein encoded by the gene, changes in amino acids, or alterations in gene expression levels.

[0029] This invention also provides the application of the temperature-sensitive male-sterile rice line described above in rice hybrid seed production, self-pollination propagation, and hybridization to cultivate new male-sterile lines.

[0030] The present invention also provides biomaterials related to the rice temperature-sensitive male sterility gene RTMF1 described in the above technical solution, characterized in that the biomaterials are any one of the following (1) to (4): (1) An expression cassette containing the rice temperature-sensitive male sterility gene RTMF1 or a partial continuous sequence as described in claim 1; (2) A recombinant vector containing the rice temperature-sensitive male sterility gene RTMF1 or a partial continuous sequence as described in claim 1; (3) Recombinant microorganisms containing the rice thermosensitive male sterility gene RTMF1 or a partial continuous sequence as described in claim 1; (4) Recombinant microorganisms containing the recombinant vector described in (2).

[0031] Example 1 RTMF1 Fertility observation of gene mutants (1) Materials: 9311 background obtained by EMS mutagenesis RTMF1 mutant ( rtmf1 ) and wild type (WT).

[0032] (2) Method: rtmf1 The mutant and WT were planted under different temperature conditions (low temperature ≤23℃, normal temperature 25℃~28℃, high temperature ≥30℃) and their fertility-related phenotypes were observed.

[0033] WT at room temperature and rtmf1 Fertility-related phenotypic results such as Figure 1 As shown, A is a schematic diagram of the plant at the heading stage, B is a schematic diagram of the spikelet at the heading stage, C is a schematic diagram of the floret, D is a schematic diagram of the anther, E and F are schematic diagrams of pollen grains stained with I2-KI, G and H are statistical graphs of the spike and seed setting rate at maturity, I and J are statistical graphs of the spike and seed setting rate after pollination with WT pollen, K and L are schematic diagrams of anther cross-sectional analysis, and MQ is a schematic diagram of the scanning electron microscopy results of anthers and pollen grains. based on Figure 1 It can be seen that, rtmf1 Male infertility occurs at normal temperatures. rtmf1 Male sterility is caused by abnormal development of the anther cuticle, tapetum, and pollen exine. Initially, results from semi-thin sections showed... rtmf1 Pollen defects are caused by delayed tapetum degradation, hindered microspore vacuolization, and abnormal undulations on the inner surface of the tapetum (based on...). Figure 1 (K and L are known). Then, scanning electron microscopy (SEM) was used to examine... rtmf1 The observations further confirmed this. rtmf1 The sterility phenotype is caused by abnormalities in the anther cuticle, the Ubsites on the inner surface of the tapetum, and the pollen exwall (based on...). Figure 1 (as can be seen from MQ). WT under high and low temperature conditions rtmf1 Fertility-related phenotypic results such as Figure 2 As shown, based on Figure 2 It can be known that: rtmf1 After high-temperature treatment during the heading stage in the field, the anthers returned to yellow and were more plump, similar to those of WT (based on...). Figure 2 (As can be seen from A and B). The I2-KI staining experiment also confirmed this. rtmf1 Most of the pollen grains were able to be stained, and the staining depth was consistent with that of WT (based on...). Figure 2 (As can be seen from C). Subsequently, regarding... rtmf1 Cross-sectional observation of the anthers revealed results similar to those of WT, where pollen grains were filled with inclusions and the tapetum layer on the inner surface of the anthers was completely degraded (based on...). Figure 2 (As can be seen from D). During the rice ripening period, rtmf1 The ears of grain are able to partially bear fruit (based on) Figure 2 As can be seen from E), this further confirms... rtmf1 The fertility was partially restored. SEM observation was used to observe... rtmf1 The anthers and pollen grains were examined, and the results showed that the cuticle of the anthers was arranged relatively normally, with spacing similar to that of WT, and the morphology of the Ubsite had been restored. The pollen grains were plump and no longer shriveled and collapsed, and the exine structure of the pollen grain surface was also significantly restored (based on...). Figure 2 (As can be seen from JN in the middle) Further rtmf1 Phenotypic analysis under low-temperature conditions in the field showed that, compared with WT, its anthers were extremely small and white, and I2-KI staining revealed no pollen production (based on...). Figure 2 (As can be seen from FH). Cross-sectional observation of the anthers showed that the pollen grains of WT were filled with inclusions, and the tapetum was completely degraded, while rtmf1 The degradation of the tapetum layer was delayed and accelerated, with most pollen grains in the anther chambers undergoing complete degradation, while some chambers showed significant degradation residues (based on...). Figure 2 (As can be seen from I). SEM observation shows that... rtmf1 The anthers show almost no pollen formation, a smooth surface without a cuticle, and no Ubsite formation on the inner surface (based on...). Figure 2 (As can be seen from OR).

[0034] In conclusion, rtmf1 The infertility phenotype responds to temperature, specifically by reducing or partially restoring fertility defects at high temperatures, while normal and low temperatures result in complete male infertility. It exhibits thermosensitive characteristics and is a thermosensitive male infertility mutant.

[0035] Example 2 RTMF1 Functional verification of genes Through MutMap analysis, determine RTMF1 To control the gene causing temperature-sensitive male infertility, RTMF1 The results of gene cloning and functional verification are as follows Figure 3 As shown, A is a schematic diagram of MutMap results analysis, B is a schematic diagram of mutation sites and gene editing sites, C1-E3 are schematic diagrams of knockout plant phenotypes, F1-F3 are schematic diagrams of complementary plant phenotypes, and G1-J2 are schematic diagrams of knockout plant phenotypes at different temperatures. Specifically: First, MutMap was used to analyze the F2 population, and the results showed that... RTMF1 A mutation at a specific site in the gene causes premature generation of a terminator, leading to premature termination of translation. Its CDS is shown in SEQ ID NO.1, and the amino acid sequence encoded by this gene is shown in SEQ ID NO.2. This gene mutation is linked to the sterility phenotype. The gene encodes a histone chaperone protein (such as...). Figure 3 (As shown in A and B). Subsequently, CRISPR / Cas9 gene editing technology was used to edit this gene in the japonica rice variety ZH11 (e.g., ...). Figure 3(As shown in Figure B). Homozygous plants were obtained at both target sites 1 and 2. Mutations in these gene-edited plants led to premature termination of mistranslation. Phenotypic observation of the gene-edited plants showed that homozygous knockout plants all exhibited significant male sterility and were unable to produce seeds later (e.g., ...). Figure 3 (As shown in C1-E3).

[0036] Using homozygous edited plants, a segregating F2 hybrid population was further constructed, confirming the co-segregation of the sterility phenotype and gene mutation. RTMF1 Key genes controlling male fertility (such as...) Figure 3 (As shown in C1-E3). Simultaneously, a gene complementation vector was constructed, and transgenic plants were obtained through genetic transformation. Phenotypic observation of the obtained complementary plants revealed that the anther and pollen staining results were consistent with the wild type, and later, fruit setting consistent with the wild type was observed (e.g., ...). Figure 3 (As shown in C1-C3, F1-F3), further confirming that RTMF1 The reproductive control function. Despite RTMF1 -ko strains produce almost no pollen, but field observations have shown that... RTMF1 -ko1-1 under low temperature conditions, anther development defects are also more severe than under normal temperature conditions (e.g. Figure 3 (As shown in G1-J2). Therefore, it can be concluded that... RTMF1 -ko fertility also responds to temperature in the Japonica rice background.

[0037] In summary, gene knockout and functional complementation have verified that... RTMF1 It refers to the gene that controls temperature-sensitive male infertility.

[0038] Example 3 Verification rtmf1 Retaining antithermally sensitive properties even in other genetic backgrounds The verification results are as follows Figure 4 As shown, Figure 4 for rtmf1 A schematic diagram of genetic analysis results in other contexts, where HT represents high temperature, NT represents normal temperature, and LT represents low temperature. Specifically: Using the maintainer XN1B as the recurrent parent, rtmf1 Transplanted into the rice XN1B background. Through hybridization and continuous backcrossing, the BC3F2 line was constructed. Near-isogenic line NIL- rtmf1 At normal temperatures, male sterility is characterized by pollen-producing flowers, while fertility recovers at higher temperatures. Furthermore, at lower temperatures, most anthers exhibit pollenless male sterility (e.g., pollenless flowers). Figure 4 (As shown in AL). This result indicates... rtmf1 It can maintain antithermally sensitive properties even under different genetic backgrounds.

[0039] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the specific implementation of the present invention. Any modifications or equivalent substitutions that do not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A rice thermosensitive male sterility gene RTMF1 and its encoded protein, characterized in that, The RTMF1 The gene sequence is shown in SEQ ID NO.1, and the encoded amino acid sequence is shown in SEQ ID NO.2; RTMF1 The mutated gene is rtmf1 Its sequence is shown in SEQ ID NO.3, and the encoded amino acid sequence is shown in SEQ ID NO.

4.

2. A rice thermosensitive male sterility gene RTMF1 The application of its mutated gene in rice production, as described in claim 1, is characterized in that... Including the use of carrying rtmf1 Genetically mutated rice male-sterile lines are used for seed production and propagation, or introduced into different rice genetic backgrounds through hybridization and backcrossing, in order to cultivate new temperature-sensitive male-sterile lines.

3. A method for cultivating a temperature-sensitive male-sterile rice line, characterized in that, Including normal rice varieties RTMF1 Gene mutation, the RTMF1 The gene sequence is shown in SEQ ID NO.1, the protein sequence is shown in SEQ ID NO.2, and it also contains a sequence that has more than 95% protein homology with it.

4. The cultivation method according to claim 3, characterized in that, Using physical, chemical, or biological techniques to improve rice varieties RTMF1 Genes can mutate, and mutations can take many forms, including those that cause premature termination of the translation of the protein encoded by the gene, changes in amino acids, or alterations in gene expression levels.

5. The application of the rice temperature-sensitive male sterile line as described in claim 3 or 4 in rice hybrid seed production, self-pollination propagation, and hybridization to cultivate new sterile lines.

6. A rice thermosensitive male sterility gene as described in claim 1 RTMF1 The related biomaterials are characterized by, The biomaterial is any one of the following (1) to (4): (1) Contains the rice temperature-sensitive male sterility gene as described in claim 1 RTMF1 Or a partially continuous sequence of expression boxes; (2) Contains the rice temperature-sensitive male sterility gene as described in claim 1 RTMF1 Or a recombinant vector containing a partially continuous sequence; (3) Contains the rice temperature-sensitive male sterility gene as described in claim 1 RTMF1 Or recombinant microorganisms with partially continuous sequences; (4) Recombinant microorganisms containing the recombinant vector described in (2).

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