Application of rice os slrl2 gene

Abstract

The application discloses application of a rice OsSLRL2 gene and relates to the technical field of plant genetic engineering. The rice OsSLRL2 gene is overexpressed in rice, and a rice strain obtained by the overexpression can reduce the plant height of the rice, improve rice quality and enhance the resistance of the rice to panicle sprouting; and the coding region sequence of the rice OsSLRL2 gene is shown as SEQ ID NO. 1. The overexpression of the rice OsSLRL2 gene in the rice can significantly improve the cooking and eating quality and the nutritional quality of rice, significantly enhance the resistance of the rice to panicle sprouting, and also has a certain inhibition on the plant height; and the application has a good application prospect in the breeding practice of cultivating new rice varieties with high yield and high quality.

Description

Technical Field

[0001] The present application relates to the technical field of plant genetic engineering, and in particular to an application of the rice OsSLRL2 gene. Background Art

[0002] Rice (Oryza sativa L.) is the staple food for more than half the world's population, and my country is the world's largest rice producer and consumer. With population growth and rising living standards, improving rice quality, yield, and resistance is crucial to ensuring food security and improving people's quality of life.

[0003] Rice quality includes processing quality, appearance quality, cooking and tasting quality, and nutritional quality. Among these, cooking and tasting quality and nutritional quality are the most important factors for consumers when purchasing rice. The main physical and chemical properties influencing rice quality include amylose content (AC), gel consistency (GC), and protein content. Appropriately reducing the AC of rice can significantly improve its cooking and tasting quality. Furthermore, rice protein content is positively correlated with its nutritional quality; the higher the protein content, the higher the nutritional value.

[0004] Pre-harvest sprouting refers to the phenomenon that the grains sprout directly on the panicle of the mother plant due to rainy weather just before the harvest of the crop. Rice panicle sprouting will cause significant losses in rice yield, as well as a decline in rice processing quality, nutritional quality, edible quality and seed vitality, and lose its commercial value. The longer the continuous high temperature and rainy weather during the rice harvest season, the more serious the damage caused by panicle sprouting. Therefore, cultivating excellent new rice varieties with good panicle sprouting resistance will help reduce the damage caused by panicle sprouting. Therefore, cloning new genes that can enhance rice panicle sprouting resistance and cultivating new rice varieties with panicle sprouting resistance will help to achieve high and stable rice yields, thereby ensuring my country's food security.

[0005] The factors that influence rice panicle germination primarily include external environmental factors and internal factors. Light and temperature are the primary environmental factors, while internal factors include seed dormancy, panicle morphology, grain water absorption characteristics, and endogenous hormones. Seed dormancy is the primary factor influencing panicle germination resistance. Some of the rice dormancy-regulating genes that have been cloned, such as OsRF1, also affect other agronomic traits of rice, impacting the application of target genes in rice breeding practices. Summary of the Invention

[0006] The present invention aims to solve at least one of the technical problems existing in the prior art or related art.

[0007] To this end, the present invention provides an application of a rice OsSLRL2 gene, wherein the rice OsSLRL2 gene is used for at least one of reducing rice plant height, improving rice quality, and enhancing rice panicle sprouting resistance; the coding region sequence of the rice OsSLRL2 gene is shown in SEQ ID NO.1.

[0008] Furthermore, the improving rice quality includes: reducing the amylose content of rice and increasing the protein content of rice.

[0009] Furthermore, the enhanced rice panicle germination resistance slightly delays rice seed germination but does not affect the final germination rate.

[0010] Furthermore, the application method is: overexpressing the rice OsSLRL2 gene in rice.

[0011] Furthermore, the rice OsSLRL2 gene is a dominantly expressed gene in rice seeds, and the protein of the rice OsSLRL2 gene is located in the nucleus of rice seeds.

[0012] Furthermore, the rice OsSLRL2 gene is overexpressed in rice using an overexpression recombinant vector; the overexpression recombinant vector comprises the rice OsSLRL2 gene, and the target vector is a plant binary expression vector.

[0013] Furthermore, the method for preparing the overexpression recombinant vector comprises:

[0014] The plant binary expression vector was double-digested with Sal I and Xba I, and the digested plant binary expression vector was ligated with a PCR amplification product containing the full-length cDNA of the rice OsSLRL2 gene using a homologous recombinase.

[0015] Furthermore, the method for amplifying the PCR amplification product containing the full-length cDNA of the rice OsSLRL2 gene comprises:

[0016] Using RNA of wild-type Nipponbare rice as a template, primer sequences SLRL2-OE-F and SLRL2-OE-R were designed for reverse transcription. The sequence of the SLRL2-OE-F is shown in SEQ ID NO.3, and the sequence of the SLRL2-OE-RF is shown in SEQ ID NO.4.

[0017] Furthermore, the method for overexpressing the rice OsSLRL2 gene in rice comprises: transforming the overexpression recombinant vector into Agrobacterium, transferring the overexpression recombinant vector into rice callus through an Agrobacterium-mediated rice genetic transformation method, and obtaining transgenic rice plants after resistance screening and tissue culture.

[0018] Furthermore, the method for overexpressing the rice OsSLRL2 gene in rice specifically comprises:

[0019] Constructing engineered bacteria: transforming the overexpression recombinant vector into Agrobacterium tumefaciens strain EHA105 by electroporation, and selecting with kanamycin and rifampicin to obtain Agrobacterium containing the overexpression recombinant vector;

[0020] Transforming rice callus and obtaining rice plants: infecting rice callus with Agrobacterium containing the overexpression recombinant vector, washing off the Agrobacterium after culture, placing the rice callus on a screening medium containing a suitable antibiotic for culture to obtain rice resistant callus, transferring the rice resistant callus to a differentiation medium for differentiation culture, transferring the differentiated seedlings to a rooting medium for culture, and obtaining rice plants after hardening.

[0021] Compared with the prior art, the present invention has at least the following beneficial effects: the present invention provides an application of the rice OsSLRL2 gene, which can reduce rice plant height, improve rice quality, and enhance rice panicle germination resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Various other advantages and benefits will become apparent to those skilled in the art by reading the detailed description of the preferred embodiment below. The accompanying drawings are only for the purpose of illustrating the preferred embodiment and are not to be considered as limiting the present application. In the accompanying drawings:

[0023] Figure 1 This is the spatiotemporal expression pattern of the OsSLRL2 gene provided in the examples of the present application.

[0024] Figure 2 The subcellular localization results of OsSLRL2 protein provided in the examples of this application.

[0025] Figure 3 Plant height analysis of OsSLRL2-overexpressing transgenic rice and wild-type controls provided in the examples of this application.

[0026] Figure 4 Comparison of grain traits between the OsSLRL2 overexpressing transgenic rice and the wild-type control provided in the examples of the present application, including grain length, grain width, and 1000-grain weight.

[0027] Figure 5 The physicochemical properties of rice provided in the examples of the present application, including apparent amylose content (AAC) and gel consistency (GC), were analyzed for the OsSLRL2 overexpressing transgenic rice and the wild-type control.

[0028] Figure 6 Comparison of rice protein content between the OsSLRL2 overexpressing transgenic rice and the wild-type control provided in the examples of this application.

[0029] Figure 7 This is a comparison of the seed germination characteristics of the OsSLRL2-overexpressing transgenic rice and the wild-type control provided in the examples of this application.

[0030] Figure 8 This is a comparison of rice panicle sprouting resistance between the OsSLRL2 overexpressing transgenic rice and the wild-type control provided in the examples of the present application. DETAILED DESCRIPTION

[0031] In order to better understand the above technical solution, the technical solution of the embodiment of the present application is described in detail below through the accompanying drawings and specific embodiments. It should be understood that the embodiment of the present application and the specific features in the embodiment are detailed descriptions of the technical solution of the embodiment of the present application, rather than limitations on the technical solution of the present application. In the absence of conflict, the embodiment of the present application and the technical features in the embodiment can be combined with each other.

[0032] An embodiment of the present invention provides an application of a rice OsSLRL2 gene, wherein the rice OsSLRL2 gene is used for at least one of reducing rice plant height, improving rice quality, and enhancing rice panicle sprouting resistance; the coding region sequence of the rice OsSLRL2 gene is shown in SEQ ID NO.1.

[0033] Improving rice quality includes reducing the amylose content and increasing the protein content. Enhancing rice panicle germination resistance means slightly delaying normal germination without affecting the final germination rate.

[0034] The application method involves overexpressing the rice OsSLRL2 gene in rice. Overexpression in rice includes overexpression in various rice tissues, such as rice seeds and leaves, with overexpression in rice seeds being the most important. The rice OsSLRL2 gene is a dominantly expressed gene in rice seeds. The protein of the rice OsSLRL2 gene is localized in the nucleus of rice seeds and can directly regulate the expression of target genes on chromosomes in the nucleus.

[0035] The applicants discovered in their research that the application of the rice OsSLRL2 gene provided by the embodiments of the present invention, through overexpression of the rice OsSLRL2 gene, can reduce the amylose content of rice, increase the protein content, and significantly enhance the rice's resistance to panicle sprouting. This provides valuable genetic resources and genetic germplasm for the cultivation of high-quality, high-yield, and panicle sprout-resistant rice varieties. This has the following beneficial effects:

[0036] (1) The rice OsSLRL2 gene is dominantly expressed in rice seeds, and its protein is mainly located in the cell nucleus; overexpression of the OsSLRL2 gene can appropriately reduce plant height to a certain extent.

[0037] (2) The rice OsSLRL2 gene is highly expressed in rice seeds and can significantly improve the cooking and nutritional quality of rice, including reducing the amylose content of rice and increasing the protein content of rice.

[0038] (3) The rice OsSLRL2 gene sequence was used to construct a recombinant vector that can be overexpressed in rice. It slightly delayed the germination of rice seeds but did not affect the final germination rate. However, it could significantly enhance the resistance of rice to panicle germination, thereby ensuring high-quality and stable rice yield. This gene has a good application prospect in the practice of breeding high-quality, high-yield and multi-resistant new rice varieties.

[0039] In a feasible embodiment, the rice OsSLRL2 gene is overexpressed in rice using an overexpression recombinant vector; the overexpression recombinant vector comprises the rice OsSLRL2 gene, and the target vector is a plant binary expression vector.

[0040] Specifically, the method for preparing the overexpression recombinant vector includes: double-digesting the plant binary expression vector with Sal I and Xba I, and connecting the digested plant binary expression vector with a PCR amplification product containing the full-length cDNA of the rice OsSLRL2 gene using a homologous recombinase.

[0041] The method for amplifying a PCR amplification product containing the full-length cDNA of the rice OsSLRL2 gene includes: using RNA of wild-type Nipponbare rice as a template, designing primer sequences SLRL2-OE-F and SLRL2-OE-R for reverse transcription, wherein the sequence of the SLRL2-OE-F is shown in SEQ ID NO. 3, and the sequence of the SLRL2-OE-RF is shown in SEQ ID NO. 4.

[0042] In a feasible embodiment, the method for overexpressing the rice OsSLRL2 gene in rice comprises: transforming the overexpression recombinant vector into Agrobacterium, transferring the overexpression recombinant vector into rice callus through an Agrobacterium-mediated rice genetic transformation method, and obtaining transgenic rice plants after resistance screening and tissue culture.

[0043] Specifically, the method for overexpressing the rice OsSLRL2 gene in rice comprises:

[0044] Construction of engineered bacteria: The overexpression recombinant vector is transformed into Agrobacterium tumefaciens strain EHA105 by electroporation, and the Agrobacterium containing the overexpression recombinant vector is obtained by screening with kanamycin and rifampicin.

[0045] Transforming rice callus and obtaining rice plants: infecting rice callus with Agrobacterium containing the overexpression recombinant vector, washing off the Agrobacterium after culture, placing the rice callus on a screening medium containing a suitable antibiotic for culture to obtain rice resistant callus, transferring the rice resistant callus to a differentiation medium for differentiation culture, transferring the differentiated seedlings to a rooting medium for culture, and obtaining rice plants after hardening.

[0046] Example 1 Application of a Rice OsSLRL2 Gene

[0047] An application of the rice OsSLRL2 gene, specifically comprising:

[0048] (1) Construction of rice OsSLRL2 gene overexpression recombinant vector pActin-SLRL2-3Flag:

[0049] The rice OsSLRL2 gene is located on rice chromosome 5 and is designated Os05g0574900 (NCBI accession number) and LOC_Os05g49930 (MSU accession number). The OsSLRL2 gene coding region CDS is 1503 bp long and lacks introns. The coding region sequence of the OsSLRL2 gene is shown in SEQ ID NO. 1. The OsSLRL2 gene encodes 500 amino acids, and the amino acid sequence encoded by OsSLRL2 is shown in SEQ ID NO. 2. The OsSLRL2 gene is contained in the pActin-SLRL2-3Flag vector, which is a plant binary expression vector.

[0050] The preparation method of the overexpression recombinant vector pActin-SLRL2-3Flag is as follows:

[0051] The plant binary expression vector pActin-3Flag was double-digested with Xba I and Sal I, and the digested vector was ligated with the PCR amplification product containing the full-length cDNA of the OsSLRL2 gene using homologous recombinase.

[0052] The PCR amplification method for the full-length cDNA containing the OsSLRL2 gene is as follows:

[0053] Using wild-type Nipponbare rice RNA as a template, the primer sequences are as follows:

[0054]

[0055] The first-strand cDNA obtained by reverse transcription is amplified using PCR to generate the target band. Following amplification, the product containing the target gene is ligated to the intermediate vector and transformed into competent E. coli cells. The cells are shaken, and single colonies are selected for enzyme digestion, identification, and sequencing. If the sequencing result is correct, the plasmid is extracted, the target gene is excised, and ligated into the vector, ultimately generating the overexpression recombinant vector pActin-SLRL2-3Flag.

[0056] (2) Construction of engineered bacteria: The pActin-OsSLRL2-3Flag vector was transformed into the Agrobacterium tumefaciens strain EHA105 by electroporation, and the Agrobacterium containing the pActin-OsSLRL2-3Flag vector was obtained by selection with kanamycin and rifampicin;

[0057] (3) Transformation of rice callus with the pActin-OsSLRL2-3Flag vector and obtaining rice regenerated seedlings: First, the mature Zhonghua 11 seeds were shelled and disinfected by soaking in 2% sodium hypochlorite solution for 1-2 h. During the disinfection period, the seeds were continuously shaken and washed, and then washed 3-5 times with sterile distilled water. The immature embryos were peeled out using pointed tweezers and a scalpel on a sterile clean bench. The embryos were transferred to an induction medium, and the grown callus tissue was used for subsequent genetic transformation. After colony PCR verification, a single Agrobacterium colony was inoculated into 4 mL of LB liquid medium containing 50 mg / L kanamycin and cultured overnight at 28°C and 250 rpm. The next day, 1% of the inoculum was transferred to 50 mL of the same medium and cultured under the same conditions for 6-8 hours until the logarithmic growth phase. The cells were centrifuged at 4000 rpm and 4°C for 5 minutes, and the cells were collected and resuspended in 10 mL of AAM liquid medium containing 100-400 μmol / L acetosyringone. The cells were then inoculated into the peeled rice callus for 20 minutes. After infection, the bacterial solution was discarded and the residual bacterial solution was removed with sterile filter paper. The callus tissue was then separated. The calli were transferred to N6D2C medium and cultured at 28°C in the dark for 3 days. After 3 days, the calli were transferred to N6D2S1 medium containing 600 mg / L cephalosporin and 25 mg / L hygromycin for the first round of screening culture. Two weeks later, the calli were transferred to new N6D2S2 medium containing 300 mg / L cephalosporin and 50 mg / L hygromycin for the second round of screening culture. The resistant calli with strong activity were then transferred to pre-differentiation and differentiation medium for culture, and the differentiated seedlings were transferred to 1 / 2MS0 rooting medium for culture. After hardening, they were transplanted to the transgenic rice experimental field.

[0058] (4) Molecular Detection of Transgenic Plants: Genomic DNA was extracted from the leaves of the transgenic rice to be tested. Forward and reverse primers were designed based on the coding sequence of the target gene OsSLRL2 in the pActin-OsSLRL2-3Flag vector and the pActin promoter sequence, respectively. These primers were used to perform molecular identification of the overexpressing rice material. The primer sequences are as follows:

[0059] sequence name sequence Sequence number pActin-F 5-TGCTGCTTCGTCAGGCTTAG-3 SEQ ID NO.5 OsSLRL2-OE-R 5-TGGAAGGAGCTGGAGAAGAT-3 SEQ ID NO.6

[0060] PCR amplification and gel electrophoresis are used to test whether the expression vector has been successfully transferred into rice. The electrophoresis band size of the PCR product should be 384bp. If the amplified fragment is the same size as the target fragment, the plant is positive; otherwise, it is negative. On the other hand, the hygromycin resistance of rice leaves is tested in vitro to further confirm the selection of positive transgenic plants. The hygromycin detection primers are as follows:

[0061] sequence name sequence Sequence number Hyg-F 5-GCTTCTGCGGGCGATTTGTGT-3 SEQ ID NO.7 Hyg-R 5-GGTCGCGGAGGCTATGGATGC-3 SEQ ID NO.8

[0062] The application of the rice OsSLRL2 gene provided in the examples of this application is further discussed below.

[0063] (I) Spatiotemporal expression pattern of the OsSLRL2 gene

[0064] Samples were collected from roots, stems, leaves, sheaths, and seeds at different developmental stages of wild-type rice plants. The samples were crushed and ground in liquid nitrogen, and total RNA was extracted. After reverse transcription, the designed OsSLRL2 gene-specific qRT-PCR primers were used to analyze the spatiotemporal expression of the OsSLRL2 gene. The results showed that OsSLRL2 is a gene that is dominantly expressed in rice seeds, and its expression level increases with the progress of seed development. Figure 1 The qRT-PCR primer sequences for the target gene OsSLRL2 and the internal reference gene Actin1 are as follows:

[0065] sequence name sequence Sequence number SLRL2-qRT-F 5-CCATGGTGCTCTCAACCAATC-3 SEQ ID NO.9 SLRL2-qRT-F 5-AATCCTTGTCATCATCGTCCTT-3 SEQ ID NO.10 Actin1-qRT-F 5-CCAAGGCCAATCGTGAGAAGA-3 SEQ ID NO.11 Actin1-qRT-R 5-AATCAGTGAGATCACGCCCAG-3 SEQ ID NO.12

[0066] (II) Subcellular localization results of OsSLRL2 protein

[0067] In order to understand the specific location of OsSLRL2 gene expression products in cells, we fused OsSLRL2 gene with GFP tag gene, constructed OsSLRL2-GFP expression vector and transferred it into rice protoplasts. After 12 hours, cells were selected and placed under laser confocal microscope to observe fluorescence signals. The results showed that OsSLRL2-GFP protein was expressed in the cell nucleus. Figure 2 The relevant vector construction primer sequences are as follows:

[0068]

[0069] (III) Investigation of plant morphology of transgenic rice plants overexpressing OsSLRL2

[0070] To understand whether overexpression of the OsSLRL2 gene affects rice plant shape, we measured the plant height of the OsSLRL2 gene overexpressing homozygous materials and their wild-type control Zhonghua 11 (ZH11) in the field. The results showed that overexpression of the OsSLRL2 gene could reduce the rice plant height to a certain extent. Figure 3 As shown, overexpression of OsSLRL2 gene has the potential to enhance rice lodging resistance.

[0071] (IV) Effects of OsSLRL2 gene overexpression on rice grain length, grain width, and 1000-grain weight

[0072] Compared with the wild-type control Zhonghua 11, overexpression of the OsSLRL2 gene had no significant effect on grain length, grain width, and 1000-grain weight of rice. Figure 4 As shown, overexpression of the OsSLRL2 gene has no effect on major yield traits such as grain shape, and is helpful to specifically improve rice quality such as cooking and eating.

[0073] (V) Investigation of OsSLRL2 gene overexpression in improving rice physical and chemical quality traits

[0074] The physical and chemical properties of rice grains of OsSLRL2 gene overexpressing rice and its wild type control Zhonghua 11 were measured. The results showed that overexpression of OsSLRL2 gene could significantly reduce the apparent amylose content of rice grains by about 5%, and slightly increase the gel consistency. Figure 5 The above results indicate that the cooking and eating quality of rice with overexpression of OsSLRL2 gene has been improved to a certain extent. In addition, the protein content of rice with overexpression of OsSLRL2 has also been significantly increased by 7% to 15% compared with the wild-type control. Figure 6 The above results indicate that overexpression of the OsSLRL2 gene also improves the nutritional quality of rice.

[0075] (VI) OsSLRL2 gene overexpression slightly delays rice seed germination

[0076] Under normal germination conditions, the germination speed of OsSLRL2 gene overexpressing rice seeds was slightly slower than that of wild type control seeds (about 12 hours), but the final germination rate still reached 100%. Figure 7 Therefore, overexpression of the OsSLRL2 gene had little effect on the normal germination of rice seeds.

[0077] (VII) Overexpression of the OsSLRL2 gene significantly enhances rice panicle sprouting resistance

[0078] The main panicles of rice from the OsSLRL2 overexpressing materials and their wild-type controls were harvested directly in the field about 30 days after flowering. High temperature and high humidity conditions were simulated in the laboratory to induce panicle germination. The results showed that the panicle germination rate of the OsSLRL2 overexpressing materials was significantly lower than that of the wild-type controls at each time point tested. The difference was particularly significant after prolonged treatment. Statistics showed that the enhanced panicle germination resistance of rice was about 60% at 6 days. Figure 8 The results of this experiment showed that overexpression of the OsSLRL2 gene can significantly enhance the panicle sprouting resistance of rice. Therefore, the OsSLRL2 gene has great application value in improving the panicle sprouting resistance of rice varieties.

[0079] It is easy for those skilled in the art to understand that, under the premise of no conflict, the above-mentioned advantageous methods can be freely combined and superimposed. The above is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present application should be included in the scope of protection of the present application. The above is only a preferred embodiment of the present application. It should be pointed out that for ordinary technicians in this technical field, several improvements and variations can be made without departing from the technical principles of the present application. These improvements and variations should also be regarded as the scope of protection of the present application.

Claims

1. An application of the rice OsSLRL2 gene, characterized in that: The rice OsSLRL2 gene is overexpressed in rice to improve at least one of rice quality and rice panicle sprouting resistance; the coding region sequence of the rice OsSLRL2 gene is shown in SEQ ID NO.1; wherein, The improved rice quality comprises: reducing the amylose content of rice and increasing the protein content of rice; The enhanced rice panicle germination resistance slightly delays the germination of rice seeds but does not affect the final germination rate.

2. The use of the rice OsSLRL2 gene according to claim 1, characterized in that: The rice OsSLRL2 gene is a dominantly expressed gene in rice seeds, and the protein of the rice OsSLRL2 gene is located in the cell nucleus of the rice seeds.

3. The use of the rice OsSLRL2 gene according to claim 1, characterized in that: The rice OsSLRL2 gene is overexpressed in rice by using an overexpression recombinant vector; the overexpression recombinant vector contains the rice OsSLRL2 gene, and the target vector is a plant binary expression vector.

4. The use of the rice OsSLRL2 gene according to claim 3, characterized in that: The method for preparing the overexpression recombinant vector comprises: The plant binary expression vector was double-digested with Sal I and Xba I, and the digested plant binary expression vector was ligated with a PCR amplification product containing the full-length cDNA of the rice OsSLRL2 gene using a homologous recombinase.

5. The use of the rice OsSLRL2 gene according to claim 4, characterized in that: The method for amplifying the PCR amplification product containing the full-length cDNA of the rice OsSLRL2 gene comprises: Using RNA of wild-type Nipponbare rice as a template, primer sequences SLRL2-OE-F and SLRL2-OE-R were designed for reverse transcription. The sequence of the SLRL2-OE-F is shown in SEQ ID NO.3, and the sequence of the SLRL2-OE-RF is shown in SEQ ID NO.

4.

6. The use of the rice OsSLRL2 gene according to claim 3, characterized in that: The method for overexpressing the rice OsSLRL2 gene in rice comprises: transforming the overexpression recombinant vector into Agrobacterium, transferring the overexpression recombinant vector into rice callus by an Agrobacterium-mediated rice genetic transformation method, and obtaining transgenic rice plants after resistance screening and tissue culture.

7. The use of the rice OsSLRL2 gene according to claim 6, characterized in that: The method for overexpressing the rice OsSLRL2 gene in rice specifically comprises: Constructing engineered bacteria: transforming the overexpression recombinant vector into Agrobacterium tumefaciens strain EHA105 by electroporation, and selecting with kanamycin and rifampicin to obtain Agrobacterium containing the overexpression recombinant vector; Transforming rice callus and obtaining rice plants: infecting rice callus with Agrobacterium containing the overexpression recombinant vector, washing off the Agrobacterium after culture, placing the rice callus on a screening medium containing a suitable antibiotic for culture to obtain rice resistant callus, transferring the rice resistant callus to a differentiation medium for differentiation culture, transferring the differentiated seedlings to a rooting medium for culture, and obtaining rice plants after hardening.

Images