CN113151295B - Rice Thermosensitive Male Sterile Gene OsFMS1 and Its Application - Google Patents

Abstract

The invention relates to the field of plant genetic engineering, in particular to a rice temperature-sensitive male sterile gene OsFMS1 and application thereof. Is characterized by no pollen formation at high temperature (about 22 ℃); while pollen is fully viable at low temperature (about less than 22 ℃), this strain is designated as fullmale-sterile (fms 1). The rice fertility gene OsFMS1 can cause the anther of rice to be aborted after over-expression or induced specific expression, can be used for cultivating a new temperature-sensitive male sterile line, can be applied to creation of the sterile line and hybrid rice seed production, and has important application value.

Description

Rice Thermosensitive Male Sterile Gene OsFMS1 and Its Application

technical field

The invention relates to the field of plant genetic engineering, in particular to a rice temperature-sensitive male sterile gene OsFMS1 and its application.

Background technique

Rice (Oryza sativa L.) is an important food crop, the staple food for half of the world's population, and an important monocot model plant. The successful creation of three-line and two-line hybrid rice has solved the problem of large-scale utilization of plant heterosis and multiplication, provided technical support for the large-scale promotion and application of hybrid rice, and made important contributions to world food security. Compared with three-line hybrid rice, two-line hybrid rice has wider restorer lines and a simpler seed production process, and it occupies an increasingly important position in hybrid rice production. The successful popularization of two-line hybrid rice benefits from the effective utilization of the photothermosensitive male sterility gene.

At present, several important photothermal-sensitive male sterility genes have been cloned in rice, such as P/TMS12 (Zhou et al, 2012); PMS3 (Ding et al, 2012); CSA (Zhang et al, 2013); TMS2 (Chueasiri et al 2014); TMS5 (Zhou et al 2014); TMS9-1 (Qi et al 2014), PMS1 (Zhou et al, 2014; Fan et al 2016), etc., but these genes cannot fully meet the requirements of breeding production As required by practice, it is still necessary to continuously discover new thermosensitive genic male sterile genes and in-depth study of their mechanism of action, which will help to quickly create excellent sterile lines, promote rice hybrid seed production and breeding, and thus increase rice yield.

In view of this, the present invention is proposed.

Contents of the invention

The present disclosure covers the following embodiments:

The isolated nucleic acid has a nucleotide sequence as shown in SEQ ID NO:1.

Isolated protein resulting from nucleic acid expression as described above.

A recombinant DNA construct comprising a nucleic acid as described above operably linked to at least one heterologous regulatory sequence.

A host cell containing a nucleic acid as described above, or a protein as described above, or transformed with a recombinant DNA construct as described above.

A method of producing transgenic rice comprising introducing a recombinant DNA construct as described above into a plant of the genus Oryza;

Wherein said nucleic acid is expressed in rice to constitute thermosensitive male sterility.

Use of the transgenic Oryza plant produced as described above for the production of rice propagation material suitable for sexual reproduction, vegetative propagation or tissue culture of regenerable cells.

The present invention found a male sterile mutant in the T-DNA insertion mutant library of the rice variety Zhonghua 11 (ZH11), which cannot produce mature pollen grains, and belongs to pollenless male sterile, and its fertility is regulated by temperature, specifically It shows that under the condition of high temperature (about 22°C), no pollen is formed; while under the condition of low temperature (about 22°C), the pollen is all fertile, so the strain is named full male-sterile (fms1) . The above-mentioned rice fertility gene OsFMS1 can be overexpressed or induced to specifically express and can abort rice anthers, which can be used to breed new temperature-sensitive male sterile lines, and can be applied to the creation of sterile lines and hybrid rice seed production, which has important Value.

Description of drawings

In order to more clearly illustrate the specific implementation of the present invention or the technical solutions in the prior art, the following will briefly introduce the accompanying drawings that need to be used in the specific implementation or description of the prior art. Obviously, the accompanying drawings in the following description The drawings show some implementations of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

Figure 1 is a schematic diagram of pollen fertility of ZH11 and Osfms1 mutants under high temperature conditions in Example 1; the wild type (WT), heterozygous (WT/osfms1) and homozygous mutants (osfms1/osfms1) in the picture A; B is the wild type Type (WT), heterozygous (WT/osfms1) and homozygous mutant (osfms1/osfms1) young panicle; C, D are flowers of wild type and homozygous mutant; E, F are wild type and homozygous mutant pollen I2/KI staining.

Fig. 2 is the light-temperature identification of osfms1 in Example 2; Zhonghua 11 (ZH11 is a wild-type control), 10 hours and 14 hours of light for 10 hours and 14 hours respectively; pollen I ₂ /KI staining.

Fig. 3 is the map of the temperature-sensitive male sterility gene OsFMS1 gene location and cloning results in Example 3; A is the OsFMS1 gene location; B is the T-DNA insertion co-segregation analysis, wherein A1-A5 is wild type, and B1-B14 is hybrid Synzygous, C1-C9 are homozygous mutants; C is a schematic diagram of the T-DNA insertion site; D is the expression of genes upstream and downstream of the T-DNA insertion site identified by semi-quantitative PCR; E is the identification of the T-DNA insertion site by qPCR Expression of downstream genes.

Figure 4 shows the overexpression of the OsFMS1 gene in an example; wherein, A is the OsFMS1 transgene overexpression plant; B is the floret of the OsFMS1 transgene overexpression plant; C is the pollen I ₂ /KI staining of the OsFMS1 transgene overexpression plant.

Detailed ways

Reference will now be made in detail to embodiments of the invention, one or more examples of which are described below. Each example is provided by way of explanation, not limitation of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. For example, features illustrated or described as part of one embodiment can be used on another embodiment to yield a still further embodiment.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field of the invention. The terms used herein in the description of the present invention are for the purpose of describing specific embodiments only, and are not intended to limit the present invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

noun definition

Before setting forth the details of the present invention, several terms used in this specification should be understood.

"Oryza" or "rice" or "rice" is a genus (Oryza) of the family Poaceae, which preferably includes the species O. sativa, further includes indica O. s. indica, japonica O. s. japonica or tropical japonica O. s. javanica subspecies.

"Agronomically superior (trait)", as used herein, refers to a genotype that possesses superior or optimal performance of a number of discernible traits that allow a producer to harvest a commercially important product. Including but not limited to seed yield, germination vigor, vegetative vigor, disease resistance, greenness, growth rate, total biomass or accumulation rate, fresh weight at maturity, dry weight at maturity, fruit yield, grain yield, Total plant nitrogen content, fruit nitrogen content, seed nitrogen content, nitrogen content in vegetative tissue, total plant free amino acid content, fruit free amino acid content, seed free amino acid content, vegetative tissue free amino acid content, total plant protein content, fruit Protein content, seed protein content, protein content in vegetative tissue, drought tolerance, nitrogen uptake, root lodging, harvest index, stalk lodging, plant height, ear height, ear length, salt tolerance, tiller number, panicle size , early seedling vigor and emergence under low temperature stress.

"Phenotype" means a detectable characteristic of a cell or organism.

A "cross" is the mating of two parental plants.

"Nucleic acid", "polynucleotide", "nucleic acid sequence", "nucleotide sequence" and "nucleic acid fragment" are used interchangeably and refer to single-stranded or double-stranded polymers of RNA and/or DNA, either The selection contains synthetic, non-natural or altered nucleotide bases. - Nucleotides (usually in their 5'-monophosphate form) are designated by the following single-letter codes: "A" for adenylic acid or deoxyadenylic acid, "C" for cytidylic acid or deoxycytidine acid, and "G" means guanylic acid or deoxyguanylic acid, corresponding to RNA or DNA, respectively; "U" means uridine acid; "T" means deoxythymidylic acid; "R" means purine (A or G); "Y" represents pyrimidine (C or T); "K" represents G or T; "H" represents A or C or T; "I" represents inosine; and "N" represents any nucleotide.

"Isolated" means a substance, such as a nucleic acid molecule and/or protein, that is substantially free of components that normally accompany or interact with it in its naturally occurring environment, or that has been removed from said Components removed. Isolated polynucleotides can be purified from host cells in which they naturally occur. Conventional nucleic acid purification methods known to the skilled artisan can be used to obtain isolated polynucleotides. The term also encompasses recombinant polynucleotides and chemically synthesized polynucleotides.

"Polypeptide", "peptide", "amino acid sequence" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. These terms apply to amino acid polymers in which one or more amino acid residues are an artificial chemical analog of the corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers. The terms "polypeptide", "peptide", "amino acid sequence" and "protein" may also include modifications including, but not limited to: glycosylation, lipid linkage and sulfation, gamma-carboxylation of glutamic acid residues, Hydroxylation and ADP-ribosylation.

"Recombinant" refers to the artificial combination of two otherwise separate segments of sequence, eg, by chemical synthesis or by manipulating the separate nucleic acid segments using genetic engineering techniques. "Recombinant" also includes reference to cells or vectors that have been modified by the introduction of heterologous nucleic acid, or cells derived from such modified cells, but does not encompass naturally occurring events (e.g., spontaneous mutation, natural transformation/ Transduction/transposition) changes to a cell or vector, such naturally occurring events as those that occur without deliberate human intervention.

"Recombinant DNA construct" refers to a combination of nucleic acid segments that do not normally occur together in nature. Thus, a recombinant DNA construct may contain regulatory and coding sequences derived from different sources, or from the same source but arranged differently than as found in nature. In some specific embodiments, the recombinant DNA construct is a plasmid or a virus. In some embodiments, the recombinant DNA construct of the present invention contains regulatory elements commonly used in genetic engineering, such as enhancers, promoters, internal ribosome entry sites (IRES) and other expression control elements (such as transcription termination signals , or polyadenylation signal and poly U sequence, etc.).

The present invention relates to an isolated nucleic acid whose nucleotide sequence is shown in SEQ ID NO:1.

The nucleic acid fragment gene claimed in the present invention also includes one or more highly homologous nucleic acid fragments shown in the nucleotide sequence SEQ ID NO: 1, and has the same highly homologous function of regulating temperature-sensitive male sterility. The equivalent body sequence of the source.

The highly homologous functional equivalent sequence includes a DNA sequence capable of hybridizing to DNA having the sequence shown in SEQ ID NO:1 under stringent conditions. The "stringent conditions" used in the present invention are well known and include, for example, hybridization at 60°C for 12 to 16 hours in a hybridization solution containing 400mM NaCl, 40mM PIPES (pH6.4) and 1mM EDTA, and then at 65°C with a mixture containing 0.1 Wash with %SDS and 0.1% SSC washing solution for 15-60 minutes.

The functional equivalent body sequence also includes at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity with the sequence shown in SEQ ID NO: 1, and has the same regulatory thermosensitive male sterility Functional gene sequences can be isolated from any plant. Wherein, the percentage of sequence identity can be obtained by known bioinformatics algorithms, including Myers and Miller algorithm (Bioinformatics, 4 (1): 11-17, 1988), Needleman-Wunsch global comparison method (J.Mol. Biol., 48(3):443-53, 1970), Smith-Waterman local alignment method (J.Mol.Biol., 147:195-197, 1981), Pearson and Lipman similarity search method (PNAS, 85 (8): 2444-2448, 1988), the algorithm of Karlin and Altschul (Altschul et al., J. Mol. Biol., 215(3): 403-410, 1990; PNAS, 90: 5873-5877, 1993). This is familiar to those skilled in the art.

The above-mentioned nucleic acid can regulate the rice temperature-sensitive male sterility function. Specifically, under high temperature (about 22°C) conditions, no pollen is formed; and under low temperature (about 22°C) conditions, all pollen is fertile. The above-mentioned rice fertility gene OsFMS1 can be overexpressed or induced to specifically express and can abort rice anthers, which can be used to breed new temperature-sensitive male sterile lines, and can be applied to the creation of sterile lines and hybrid rice seed production, which has important Value.

The present invention also relates to isolated proteins expressed from nucleic acids as described above.

The present invention also relates to recombinant DNA constructs comprising a nucleic acid as described above operably linked to at least one heterologous regulatory sequence.

The invention also relates to host cells containing a nucleic acid as described above, or a protein as described above, or transformed with a recombinant DNA construct as described above.

The nucleic acid fragments provided by the present invention can be inserted into plasmids, cosmids, yeast artificial chromosomes, bacterial artificial chromosomes or any other vectors suitable for transformation into host cells. Preferred host cells are bacterial cells, especially for cloning or storing polynucleotides, or for transforming plant cells, such as Escherichia coli, Agrobacterium, Agrobacterium tumefaciens and Agrobacterium rhizogenes.

The present invention also relates to a method for producing transgenic rice, comprising introducing a recombinant DNA construct as described above into a plant of the genus Oryza;

Wherein said nucleic acid is expressed in rice to constitute thermosensitive male sterility.

In some embodiments, the recombinant DNA construct is introduced into rice by host cell introduction as described above.

The methods of the present invention can also directly introduce the nucleic acid into plants to produce transgenic rice, prepared using transformation methods known to those skilled in the field of plant biotechnology. Any method can be used to transform the recombinant expression vector into plant cells to produce the transgenic plants of the invention. Transformation methods can include direct and indirect transformation methods. Suitable direct methods include liposome-mediated transformation, introduction using a gene gun, electroporation, and pollen tube passage, among others.

In some embodiments, the recombinant DNA construct is introduced into rice by hybridization.

In some embodiments, the method includes:

1). Determining whether the first Oryza plant has the nucleic acid fragment shown in SEQ ID NO: 1;

2). Optionally verifying the temperature-sensitive male sterile phenotype of the first Oryza plant;

3). crossing said first Oryza plant with a second Oryza plant to produce progeny plants;

4). Optionally use the progeny plants described in step 3) as raw materials, and repeat steps 1)-3) 2-10 times to produce other progeny plants.

In some embodiments, the second Oryza plant is an agronomically elite variety.

The present invention also relates to the use of a transgenic Oryza plant produced by the method described above for the production of rice propagation material, wherein the propagation material is suitable for sexual reproduction, vegetative propagation or tissue culture of regenerable cells;

In some embodiments, said reproductive material suitable for sexual reproduction is selected from microspores, pollen, ovaries, ovules, embryo sacs and egg cells.

In some embodiments, said propagation material suitable for vegetative propagation is selected from cuttings, roots, stems, cells, protoplasts.

In some embodiments, said propagation material suitable for tissue culture of regenerable cells is selected from the group consisting of leaves, pollen, embryos, cotyledons, hypocotyls, meristematic cells, roots, root tips, anthers, flowers, seeds and stem.

Embodiments of the present invention will be described in detail below in conjunction with examples.

The rice variety Zhonghua 11 used in the examples is a standard variety.

Phenotype identification and genetic analysis of embodiment 1 fms1 mutant

The OsFMS1 mutant was found in the T1 generation of the Zhonghua 11T-DNA insertion mutant library. The homozygous mutants were semi-dwarfed, with dark green leaves, half-covered panicles (Fig. 1A, B), and no fertile seeds at maturity. The seed setting rate of heterozygous plants was 31.1±8.4%, which was significantly lower than that of wild type (85.4±7.2%). When the florets were observed under a dissecting microscope, the stigma of the mutant florets developed normally, the number and structure of floral organs were normal, but the filaments were long and thin, and the anthers shrank and turned white (Fig. 1C, D). The anthers of florets before flowering were stained with 1% I2/KI, and it was found that there were no mature pollen grains in the mutant anthers (Fig. 1E, F).

In the T1 generation lines, wild type: semi-sterile: complete sterile plants = 5:14:9, conforming to the segregation relationship of 1:2:1 (χ ² _c =0.43<χ ² _0.05 =3.84). Harvest the wild-type and semi-sterile plants and plant the T2 generation, and the wild-type will no longer segregate; the semi-sterile plants will segregate into wild-type, semi-sterile and complete sterile plants, and the segregation ratio is 1:2:1 separation relationship.

Table 1 Plant Separation Situation

The homozygous mutants were used as parents to cross with the original parent Zhonghua 11 respectively. When the homozygous mutant is used as the male parent, no seeds are formed; when the homozygous mutant is used as the female parent, seeds can be obtained normally, F1 is semi-sterile, F2 generation wild type: semi-sterile: completely sterile plants = 15 :32:12, conforming to the separation relationship of 1:2:1 (χ ² _c =0.46<χ ² _0.05 =3.84).

Based on the above phenotype and genetic analysis results, it was determined that the OsFMS1 mutant was apollen male sterile and controlled by a pair of incompletely dominant nuclear genes.

Example 2 Photothermal identification of fms1 mutants

The fms1 mutant showed no pollen male sterility and no mature seeds under the long-day high-temperature conditions in Hangzhou summer (heading around August 10, long-day high temperature), and matured around October 1 (low temperature, short day). The spikes are subtracted, and the newly drawn young spikes can bear fruit normally. The rice piles and harvested seeds of OsFMS1 homozygous plants are sent to Hainan for planting. Around March 10 (short day, low temperature) the earing of OsFMS1 mutations returns to normal, and can be harvested. Normal fruiting suggested that the mutant might be controlled by photoperiod or temperature. Use the light incubator to set 4 temperature and light combinations. The culture conditions are: high temperature long day: 28°C + 14h light; high temperature short day 28°C + 10h light; low temperature long day 22°C + 14h light; low temperature day 22°C +10h light. Investigating the pollen fertility of plant spikelets, it was found that ZH11 had normal and fertile pollen in all treatments, while OsFMS1 showed normal and fertile pollen only under the low temperature condition of 22°C, and showed no pollen phenotype under high temperature condition (Figure 2) . It was confirmed that the pollen fertility of the OsFMS1 mutant was mainly regulated by temperature, and OsFMS1 was a temperature-sensitive genic sterile mutant.

Example 3 Location and cloning of OsFMS1 gene

The OsFMS1 mutant was used as the female parent and the indica rice variety Longtefu B was used as the male parent to prepare the mapping population, and the individuals whose phenotype was extremely recessive were used as the mapping population for gene mapping. Using 192 pairs of polymorphic SSR primers covering 12 rice chromosomes, the mutated gene was initially located, and the mutated gene was located on chromosome 4, and then the candidate gene was further located using SSR primers and newly designed STS markers within a physical distance of 103 kb (Fig. 3A).

The SSR primer and STS primer are as follows:

tag name F(5'-3') R(5'-3') RM3288 CAATCTGGAGGCACTGTCACG AGTGACAAGATGAAGCCAACAGC RM6089 CGATGGCCAGCGTGATCTCC CCACCGAATCGAATAACCACAAGC RM3474 ACCTCACCTTTCCCTCGATTGG GTTGGTTGCTTCCTCCCATACG RM3276 ACAGGTCGATCTCGATGAACTCC CTCTTCTCCGTCTCGACTCTTCC RM3836 CGGAATCACCAAATTTTCTCTCTCAGC CGCAAGAAACGGAAACGAAACC 4-1 AGTGCTGTCAGAGAGAAAGCAATCC GGAGCCATCGAGTGTCATCACC 4-4 ACTTTGCTCTGAACTTGCAGTGG GCTAGCTGCTATCAGGATTCACG 4-11 TTGGAAGCCAACTCAAGTTACCC GAAGCAGGGATGTGAGAGATGG 4-15 AGACCATAATGCCACAATCC TTGATTTGTGCTCTTCCCAG 4-18 CAGAAGAACAAACTCCACAG AAATCGATCCTCTGCAGTGC

At the same time, in order to determine whether the mutant phenotype co-segregates with T-DNA, that is, whether the mutant is an insertion mutation. All single plants of the T1 generation were selected, and the HPT gene in the T-DNA was used for PCR amplification. It was found that both heterozygous and homozygous mutants could amplify the target band, while the wild type had no amplified product. In the T2 generation, all wild-type strains had no amplified products, and in the strains that continued to segregate, wild-type had no amplified products, and both heterozygous and homozygous mutants could amplify the target band (Fig. 3B) . It indicated that T-DNA co-segregated with the mutant phenotype, and the mutant was probably caused by T-DNA insertion.

The flanking sequence of the T-DNA insertion site was isolated by TAIL-PCR technology, and sequenced by the rice genome annotation database TIGR (http://rice.plantbiology.msu.edu/) and RiceGAAS (http://ricegaas.dna .affrc.go.jp/rgadb/) analysis found that the T-DNA was inserted into the above-mentioned finely positioned 103kb interval, located between two genes in the same direction (Figure 3C)

Combined with the results of gene mapping, two genes upstream and downstream of the T-DNA insertion site were selected for expression analysis. The cDNA of the young panicle from the third stage of development to the third stage of V was selected for semi-quantitative PCR. Through the analysis of the experimental results, it was found that the expression of the ORF1 gene (LOC_Os04g50030) downstream from the T-DNA insertion site was significantly increased, while the expression of the other three candidate genes had no significant difference (Figure 3D). qRT-PCR was used to further detect the expression level changes of the OsFMS1 gene in each period, and it was found that the expression level of the OsFMS1 gene increased significantly in each period ( FIG. 3E ). It indicated that ORF1 gene (LOC_Os04g50030) was the candidate gene of OsFMS1. Sequencing analysis of the amplified full-length ORF product, and through database comparison analysis, it was found that the OsFMS1 gene has no introns and encodes an AT-HOOK transcription factor. At present, there is no report on the cloning and functional research of this gene.

The primer sequences used in the RT-PCR are as follows:

Example 4 Overexpression of OsFMS1 gene

A UBI promoter-driven OsFMS1 gene overexpression vector was constructed to transform wild-type Zhonghua 11 plants, and the amplification primers were: OsFMS1-BamHI: (5'-TGCTGGATCCGTACATGAGCTAGTGGAG-3'); OsFMS1-KpnI: (3'-CCAGGTACCCTTGCGATGTCTCCTTTC-5' ), the amplification primers contain two restriction sites, BamHI and KpnI, for connection to the pCAMBIA1301-Ubi/Nos vector. The reaction conditions were: 94°C pre-denaturation for 7 minutes; 94°C/1min, 60°C/30s, 72°C/3min, 35 cycles; 72°C extension for 7min. The PCR reaction system is:

The amplified product was identified and recovered, digested with BamHI and KpnI, and connected to the pCAMBIA1301-Ubi/Nos vector that was also digested with BamHI and KpnI to construct the pCAMBIA1301-Ubi-OsFMS1 vector, and transformed into Huahua by Agrobacterium-mediated method 11 wild-type plants. The obtained transgenic seedlings were identified.

The transformed plants had normal fertility under low temperature conditions, and under high temperature conditions, the transformed plants showed similar phenotypes to OsFMS1 (Fig. 4A), anther characteristics and pollen fertility were also similar to OsFMS1 phenotypes (Fig. 4B). The anthers of florets before flowering were stained, and it was found that no mature pollen grains were formed in the anthers of overexpression plants (Fig. 4C). Therefore, it was determined that LOC_Os04g50030 was the OsFMS1 gene.

The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, should be considered as within the scope of this specification.

The above-mentioned embodiments only express several implementation modes of the present invention, and the descriptions thereof are relatively specific and detailed, but should not be construed as limiting the patent scope of the invention. It should be noted that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

Claims (8)

1. A method for producing transgenic rice, comprising introducing a recombinant DNA construct into rice; The recombinant DNA construct comprises a nucleic acid with a nucleotide sequence operably linked to at least one heterologous regulatory sequence as shown in SEQ ID NO: 1; Wherein said nucleic acid is expressed in rice to constitute thermosensitive male sterility. 2. The method according to claim 1, wherein said recombinant DNA construct is introduced into rice by gene gun-mediated transformation method, pollen tube passage method or liposome transformation method. 3. The method according to claim 1, wherein said recombinant DNA construct is introduced into rice by hybridization. 4. The method of claim 3, comprising: 1). Determining whether the first rice plant has the nucleic acid fragment shown in SEQ ID NO: 1; 2). Verifying the thermosensitive male sterile phenotype of the first rice plant; 3). Crossing said first rice plant with a second rice plant to produce progeny plants; 4). Using the progeny plants described in step 3) as raw materials, repeat steps 1)-3) 2-10 times to produce other progeny plants. 5. The method of claim 4, the second rice plant being an agronomically elite variety. 6. Use of the transgenic rice produced by the method according to any one of claims 1 to 5 for producing rice propagation material, wherein the propagation material is suitable for sexual reproduction or tissue culture of regenerable cells. 7. The use according to claim 6, the propagating material suitable for sexual reproduction is selected from the group consisting of microspores, pollen, ovary, ovules, embryo sacs and egg cells. 8. purposes according to claim 6, the described propagating material that is suitable for the tissue culture of regenerable cell is selected from leaf, pollen, embryo, cotyledon, hypocotyl, meristem cell, root, anther, flower, Seeds and stems.

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