A gene involved in the rice pal-dependent salicylic acid synthesis pathway and application thereof

Abstract

The application belongs to the field of plant genetic engineering, and discloses a gene involved in a PAL-dependent SA synthesis pathway of rice and application thereof, wherein the nucleotide sequence of OSD2 is SEQ ID NO:1, the nucleotide sequence of a protein coding region is SEQ ID NO:2, and the amino acid sequence of a protein coding protein is SEQ ID NO:3; the OSD2 gene is knocked out in the background of Zhonghua No. 11 through CRISPR / Cas9, a mutant OSD2 is obtained, in which the SA content of rice leaves is reduced to 15% of the wild type, phenotype analysis and hormone determination are carried out, a pUbi-OSD2-pMDC32 overexpression vector is constructed, and OSD2 overexpression plants are created. The OSD2 functional loss leads to a significant decrease in the SA content of rice, and the rice is more sensitive to X. oryzae. The OSD2 overexpression has a strong resistance to X. oryzae.

Description

Technical Field

[0001] This invention belongs to, but is not limited to, the field of plant genetic engineering technology, and particularly relates to a gene involved in the PAL-dependent salicylic acid synthesis pathway in rice and its application. Background Technology

[0002] Salicylic acid (SA) is an important plant hormone that participates directly or indirectly in plant growth, development, and stress responses through crosstalk with other hormones. SA is one of many phenolic compounds synthesized by plants, participating in multiple life processes such as seed germination, cellular respiration, cell growth, stomatal opening and closing, flowering, and senescence. Furthermore, SA is a key signal regulating heat production and disease resistance.

[0003] Plant ammonia synthesis primarily occurs through two secondary metabolic pathways originating from shikimic acid: the isochorismate synthase (ICS) pathway and the phenylalanine ammonia-lyase (PAL) pathway. Plants synthesize SA via both the ICS and PAL pathways, and the SA signaling pathway plays a crucial role in plant disease resistance. In rice, ICS1 mainly participates in chlorophyllin biosynthesis but not in SA biosynthesis. The PAL pathway in rice is not fully understood. Studies have shown that loss of function of the gene AIM1 (ABNORMALINFLORESCENCE MERISTEM 1), which is involved in the PAL-dependent SA synthesis pathway, reduces the endogenous SA content in rice to about 10% of the wild-type level, indicating that the PAL pathway is the main pathway for SA synthesis in rice. In barley inoculated with *Pseudomonas syringae* pv. *syringae*, endogenous salicylates (SA) increased during pathogen infection. However, in rice, SA levels did not increase during infection with *Pseudomonas syringae*, *Magnaporthe grisea*, or *Rhizoctonia solani*. Salicylate hydroxylasase (NahG) can block SA accumulation in the body. In NahG transgenic Arabidopsis, SA levels significantly increased after *Pto DC3000* pathogen invasion, indicating that SA plays an important role in Arabidopsis disease resistance. Rice expressing NahG showed a significant increase in ROS content after *Bacillus oryzae* invasion, suggesting that decreased SA levels lead to increased susceptibility to *Bacillus oryzae*. Mutations in OsS5h increase SA content in rice and enhance its resistance to bacterial blight. Overexpression of OsS5h reduces endogenous SA in rice and increases its susceptibility to bacterial blight, indicating a positive correlation between SA content and rice resistance to bacterial blight. Recent studies suggest that SA may also mediate defense signaling in wheat. In summary, these results indicate that, at least in some monocotyledonous plants, SA is a signal of the defense response, and endogenous SA content is positively correlated with plant disease resistance.

[0004] Rice is the staple crop for more than half of the world's population and is also a monocotyledonous model plant with a high basal salicylic acid (SA) level. Rice production is frequently affected by various field diseases, the most destructive of which is bacterial bacterial blight caused by Xanthomonas oryzae. Bacterial blight can affect rice growth, development, and reproduction. Therefore, cultivating high-yielding and stable-yielding new crop varieties is of great significance for ensuring food security.

[0005] Based on the above analysis, the urgent technical problems that need to be solved in the existing technology are:

[0006] Rice cannot accumulate large amounts of saprophytic acid (SA) after pathogen invasion, which prevents the plant from effectively utilizing SA to resist external pathogen invasion. In plant disease resistance improvement, any increase in resistance is often accompanied by changes in growth and development and a decrease in yield. Therefore, it is crucial to improve rice's resistance to pathogens while ensuring stable yields. Summary of the Invention

[0007] To address the problems existing in the prior art, this invention provides a gene involved in the PAL-dependent salicylic acid synthesis pathway in rice and its application.

[0008] This invention is achieved by providing a gene involved in the PAL-dependent salicylic acid synthesis pathway in rice, namely OSD2 (Oryza Sativa SA-Deficient gene 2), with the nucleotide sequence SEQ ID NO: 1.

[0009] Furthermore, the salicylic acid synthesis gene further includes a DNA sequence with more than 90% homology to the DNA sequence shown in SEQ ID NO: 1.

[0010] Furthermore, the salicylic acid synthesis gene further includes one or more alleles or gene derivatives with base alterations generated through base substitution, deletion, or addition.

[0011] Furthermore, the salicylic acid synthesis gene further comprises a DNA molecule capable of hybridizing with the DNA sequence shown in SEQ ID NO: 1 under stringent conditions.

[0012] Another object of the present invention is to provide a protein encoded by the rice salicylic acid synthesis gene, wherein the nucleotide sequence of the encoded protein is SEQ ID NO: 2.

[0013] Furthermore, the encoded protein amino acid sequence is SEQ ID NO: 3;

[0014] The encoded protein further includes an amino acid sequence that has more than 90% homology with the amino acid sequence shown in SEQ ID NO: 3;

[0015] The encoded protein further includes proteins and protein analogs that have undergone one or more amino acid alterations based on the amino acid sequence shown in SEQ ID NO: 3 through amino acid substitution, deletion, or addition.

[0016] The encoded protein further includes a fusion protein formed by linking the protein shown in SEQ ID NO: 3 with other tagged proteins.

[0017] Another objective of this invention is to provide a plant genetic transformation vector constructed using the SA synthetic gene, wherein the plant genetic transformation vector includes an expression vector that upregulates OSD2; the expression vector that upregulates OSD2 includes a fusion expression vector constructed using a recombinant promoter or a tissue-specific promoter.

[0018] The plant genetic transformation vector further comprises: a DNA sequence containing the DNA sequence shown in SEQ ID NO: 1, or a DNA sequence having more than 90% homology with the DNA sequence shown in SEQ ID NO: 1, or an allele or gene derivative with one or more base changes generated by base substitution, deletion, or addition of the nucleotide sequence shown in SEQ ID NO: 1, or a DNA molecule that can hybridize with the DNA sequence shown in SEQ ID NO: 1 under strict conditions.

[0019] Furthermore, the plant genetic transformation vector further includes an OSD2 expression upregulation or downregulation vector, and downregulates the expression of the gene shown in SEQ ID NO: 3 through CRISPR / Cas9 technology, T-DNA insertion technology, EMS mutagenesis, RNA interference technology, and gene silencing technology;

[0020] The plant genetic transformation vector upregulates or downregulates the expression level or activity of the protein shown in SEQ ID NO: 3 using relevant protein regulators.

[0021] Another objective of this invention is to provide a recombinant bacterium, plant callus, and cell line expressed using the aforementioned plant genetic transformation vector.

[0022] Another objective of this invention is to provide a method for regulating the resistance of rice leaves to bacterial blight. The method specifically includes: knocking out the OSD2 gene, which leads to a decrease in the salinity (SA) of rice leaves and sensitivity to bacterial blight; and overexpressing the OSD2 gene, which increases the resistance of rice to bacterial blight.

[0023] Based on the above technical solutions and the technical problems solved, the advantages and positive effects of the technical solution to be protected by this invention are as follows:

[0024] The present invention provides the rice SA synthesis gene OSD2 and its application, demonstrating that rice OSD2 can regulate rice SA synthesis, create its overexpression materials, and increase rice disease resistance. Attached Figure Description

[0025] Figure 1This invention provides the identification of the OSD2 gene involved in the SA biosynthesis pathway in rice: A. Location of the Cas9-induced mutagenesis target motif (red arrow) within the OSD2 gene coding region (orange box); B. Sanger sequencing peak diagram of the osd2 mutant; C. Encoded amino acid sequence information of wild-type and osd2 mutant; D. Phenotype of wild-type and osd2 mutant plants at the heading stage, scale bar 10 cm; E. SA and SAG content in leaves of 14-day-old wild-type and osd2 mutant plants; F. Phenotype and lesion length statistics of wild-type and osd2 mutant plants 14 days after Xoo inoculation, scale bar 3 cm. Statistical analysis was performed using a two-tailed student's t-test, ***p < 0.001. Data are expressed as mean ± standard deviation, n = 4 (E) and n = 20 (F) biologically independent samples.

[0026] Figure 2 The following are the expression patterns of OSD2 provided in this embodiment of the invention: A. The expression of the OSD2 gene in the roots, stems, leaves, and spikes of 70-day-old Zhonghua 11 plants. The expression levels of the gene were normalized with the internal reference gene OsUBQ5. B. The expression of the OSD2 gene in the leaves of Zhonghua 11 plants inoculated with Xoo at 0, 12, 24, 48, and 72 hours. The expression levels of the OSD2 gene after inoculation with Xoo were normalized with the corresponding control group. Statistical analysis was performed using a two-tailed student's t-test. *p < 0.05, ***p < 0.001. Data are expressed as mean ± standard deviation, n = 3 (A and B) biologically independent samples.

[0027] Figure 3 This invention provides the subcellular localization of OSD2 in rice protoplasts, with mCherry-AIM1 used as a peroxisome localization marker, and a scale bar of 10 μm.

[0028] Figure 4 The OSD2 gene overexpression provided in this embodiment of the invention enhances the disease resistance of rice: A. Expression level of pUbi-OSD2-pMDC32 / ZH11 overexpression material at the tillering stage in the field; B. Field-grown plants of Zhonghua 11 and pUbi-OSD2-pMDC32 / ZH11 inoculated with Xoo phenotype at the tillering stage, scale bar 3 cm; C. Statistical analysis of lesion length on leaves of Zhonghua 11 and three OSD2 overexpression lines 14 days after inoculation with Xoo strain. Statistical analysis was performed using the two-tailed Student's t-test, ***p <0.001. Data are expressed as mean ± standard deviation, n = 3 (A) and n = 20 (C) independent biological replicates. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0030] This invention provides a gene involved in the PAL-dependent salicylic acid synthesis pathway in rice, the gene being OSD2, with the nucleotide sequence SEQ ID NO: 1.

[0031] The salicylic acid synthesis gene further includes a DNA sequence with more than 90% homology to the DNA sequence shown in SEQ ID NO: 1.

[0032] The salicylic acid synthesis gene further includes one or more alleles or gene derivatives with altered bases produced by base substitution, deletion, or addition.

[0033] The salicylic acid synthesis gene further includes a DNA molecule capable of hybridizing with the DNA sequence shown in SEQ ID NO: 1 under stringent conditions.

[0034] This invention provides a protein encoded by the rice salicylic acid synthesis gene, wherein the encoded protein has the nucleotide sequence SEQ ID NO: 2.

[0035] The encoded protein amino acid sequence is SEQ ID NO: 3;

[0036] The encoded protein further includes an amino acid sequence that has more than 90% homology with the amino acid sequence shown in SEQ ID NO: 3;

[0037] The encoded protein further includes proteins and protein analogs that have undergone one or more amino acid alterations based on the amino acid sequence shown in SEQ ID NO: 3 through amino acid substitution, deletion, or addition.

[0038] The encoded protein further includes a fusion protein formed by linking the protein shown in SEQ ID NO: 3 with other tagged proteins.

[0039] This invention provides a plant genetic transformation vector constructed using the SA synthetic gene, wherein the plant genetic transformation vector includes an expression vector that upregulates OSD2; the expression vector that upregulates OSD2 includes a fusion expression vector constructed using a recombinant promoter or a tissue-specific promoter.

[0040] The plant genetic transformation vector further comprises: a DNA sequence containing the DNA sequence shown in SEQ ID NO: 1, or a DNA sequence having more than 90% homology with the DNA sequence shown in SEQ ID NO: 1, or an allele or gene derivative with one or more base changes generated by base substitution, deletion, or addition of the nucleotide sequence shown in SEQ ID NO: 1, or a DNA molecule that can hybridize with the DNA sequence shown in SEQ ID NO: 1 under strict conditions.

[0041] The plant genetic transformation vector further includes an OSD2 expression upregulation or downregulation vector, which downregulates the expression of the gene shown in SEQ ID NO: 3 through CRISPR / Cas9 technology, T-DNA insertion technology, EMS mutagenesis, RNA interference technology, and gene silencing technology.

[0042] The plant genetic transformation vector upregulates or downregulates the expression level or activity of the protein shown in SEQ ID NO: 3 using relevant protein regulators.

[0043] This invention provides a recombinant bacterium, plant callus, and cell line expressed using the plant genetic transformation vector.

[0044] This invention provides a method for regulating the resistance of rice leaves to bacterial blight. The method specifically includes: knocking out the OSD2 gene, which leads to a decrease in the salinity (SA) of rice leaves and sensitivity to bacterial blight; and overexpressing the OSD2 gene, which increases the resistance of rice to bacterial blight.

[0045] This invention knocks out the Os10g0503300 gene using CRISPR / Cas9 technology and verifies its function. Through research on its basic biological functions, it was found that it can participate in the PAL-dependent SA synthesis pathway and participate in rice disease resistance.

[0046] Required experimental material: Wild-type japonica rice Zhonghua 11 (ZH11)

[0047] Essential primer information for gene editing: The ZH11-background rice osd2 mutant was created using CRISPR / Cas9 gene editing technology. Specific primers were GGCAGAAGTGCTCGAGCCTCACGT and AAACACGTGAGGCTCGAGCACTTC as the knockout target. After constructing the relevant vector, it was sent to Wuhan Boyuan Biotechnology Co., Ltd. for transgenic processing. Identification primers: TGGTGCCCTTGGTGTCC and GGGCTGAGGTTCCACATC were used for genome cloning. The PCR products were sent to the sequencing department of Hangzhou Qingke Biotechnology Co., Ltd. for sequencing.

[0048] Experimental methods:

[0049] SA extraction and determination

[0050] 1) Take approximately 50 mg of leaf tissue, quickly freeze it in liquid nitrogen, and store it in a -80°C freezer until use;

[0051] 2) Finely grind using a cryogenic grinder, and extract with 500 μL of methanol and internal standard (D6-SA) in a rotary mixer at 4°C for at least 4 hours.

[0052] 3) Centrifuge at 12000 g for 10 minutes at 4℃, and extract the remaining mixture once at 4℃ with 500 μL of 100% methanol for more than 1 hour.

[0053] 4) Centrifuge at 12000 g for 10 minutes at 4℃, mix the supernatant and dry with nitrogen; then dissolve the residue in 300 μL of 30% methanol and quantify SA and SAG using an ExionLC (AB SCIEX) high performance liquid chromatography (HPLC) instrument and a QTRAP 5500 mass spectrometer (AB SCIEX).

[0054] White leaf blight inoculation experiment:

[0055] 1) Introduce rice bacterial blight virus P6 (Xanthomonas oryzae pv. oryzae (Xoo) Philippinestrain P6 (PXO99) A The strain was streaked on modified medium and incubated at 28°C for 2–3 days;

[0056] 2) Streak the strain again to activate it;

[0057] 3) Resuspend the bacterial cells in sterile water to OD. 600 = 1.0, and immediately used for plant inoculation;

[0058] 4) Inoculate the materials grown in the field or in the culture room when they reach the tillering stage. One week before inoculation, change the conditions of the culture room to 30 / 28℃, 12 h / 12 ​​h, and 80% air humidity.

[0059] 5) For materials grown in the field, disease incidence should be recorded and photographed 14 days after inoculation, and for materials grown in the culture room, disease incidence should be recorded and photographed 21 days after inoculation.

[0060] Rice protoplast preparation and transformation:

[0061] 1) Prepare wild-type rice seedlings that have been hydroponically grown for 2-3 weeks;

[0062] 2) Use a sharp razor to cut the plant material into small pieces 0.5–1 mm wide and place them in an enzymatic hydrolysate for digestion;

[0063] 3) Vacuum treatment for 10 minutes (2–3 times), 20–22℃, and stand still in the dark for 3–4 hours;

[0064] 4) Gently vortex the enzyme solution to release protoplasts, and the solution will gradually turn green;

[0065] 5) Filter the protoplast enzyme solution into a 50 mL conical centrifuge tube using a 35-75 μm nylon membrane (wall-adherent filtration);

[0066] 6) Centrifuge at 100 g, speed 1, for 3 minutes. Centrifuge the protoplasts to the bottom of the centrifuge tube. Repeat 3–5 times to collect more protoplasts.

[0067] 7) Use W5 solution to suspend the protoplasts at the bottom, centrifuge at 100 g for 3 minutes to transfer the protoplasts to the bottom;

[0068] 8) Centrifuge at 100 g for 3 minutes at a speed of 1 to bring the protoplasts to the bottom and suspend them in MMG solution;

[0069] 9) Examine the protoplast quality under a microscope, place on ice for 15–30 minutes, and allow for transformation;

[0070] 10) Add 10 μL of plasmid (concentration of 1,500 ng / μL) to 100 μL of protoplasts and mix gently.

[0071] 11) Add 110 μL of PEG4000, mix gently, and incubate at room temperature for 20 minutes;

[0072] 12) Then add 1 mL of W5 solution and mix gently;

[0073] 13) 100 g, centrifuge at a speed of 1 for 3 minutes, and remove the supernatant;

[0074] 14) Add 1 mL of W5 solution, 100 g, centrifuge at speed 1 for 3 minutes, and wash the protoplasts;

[0075] 15) Discard the supernatant, add 1 mL of W5 solution, and incubate at 20°C in the dark for 12–16 hours to observe subcellular localization.

[0076] PCR primer information and reaction conditions:

[0077] Carrier construction and transformation:

[0078] Table 1 Seamless Cloning Reaction System

[0079]

[0080] 1) Add the target fragment and the vector according to Table 1, vortex mix, and react at 37°C for 30 minutes;

[0081] 2) After the reaction is complete, 5 µL of the ligation product is transferred into DH5α competent cells, mixed well, and incubated on ice for 20–30 minutes, gently tapping the container every 10 minutes to mix.

[0082] 3) Heat shock in a 42℃ water bath for 90 seconds, then immediately place on ice and incubate for 3–8 minutes, add antibiotic-free LB culture and activate by shaking in a 37℃ shaker for 45–60 minutes;

[0083] 4) After activation, centrifuge at low speed, discard excess culture medium, resuspend the bacterial cells, and spread them onto LB solid medium containing the corresponding carrier resistance. Finally, invert the spread and dried plates in a 37°C incubator overnight.

[0084] After transforming the recombinant vector plasmid into E. coli, the successful construction and transformation of the vector were verified by colony PCR, plasmid PCR, and plasmid enzyme digestion.

[0085] 1) Colony PCR Identification: Prepare the PCR reaction system in advance, pick a single colony from the previous inverted culture and add it to the system. Amplify the target fragment by PCR. If the size of the amplified band is correct, the vector construction and transformation are considered successful. The specific reaction system is shown in Table 2.

[0086] Table 2. PCR reaction system 2)

[0088] 3) PCR reaction conditions are shown in Table 3:

[0089] Table 3. PCR reaction procedure

[0090]

[0091] 3) Expand the colony system that was identified as positive by colony PCR, extract plasmids and perform plasmid PCR for further verification. The PCR reaction system and conditions are the same as above.

[0092] 4) After the reaction is complete, agarose gel electrophoresis is performed. If the band size is correct, the recombinant vector has been successfully constructed and sent to the sequencing department of Hangzhou Qingke Biotechnology Co., Ltd. for sequencing.

[0093] Example 1: Identification of OSD2 gene involved in the rice SA biosynthesis pathway

[0094] To elucidate the PAL pathway in SA biosynthesis, we knocked out the Os10g0503300 gene using CRISPR / Cas9 technology and verified its function. Os10g0503300, predicted to be benzyl alcohol O-benzoyltransferase, was named OSD2. This gene is 1428 bp in length, intronless, and encodes a protein of 475 amino acids. Knockout using CRISPR / Cas9 yielded three independent homozygous lines. In line osd2-1, an A base was inserted at position 334 of the ATG, resulting in a frameshift mutation starting at position 112. In line osd2-2, a T base was inserted at position 334 of the ATG, resulting in a frameshift mutation starting at position 112. In line osd2-3, a C base was inserted at position 335 of the ATG, resulting in a frameshift mutation starting at position 112. Figure 1 (A–C in the original text). The osd2 mutant showed no significant difference from WT during growth ( Figure 1 The D in the sample showed that the endogenous SA and SAG contents in the 14-day-old leaves decreased to less than 15% of those in the wild type. Figure 1 The presence of E in the data indicates that the OSD2 gene plays a crucial role in the synthesis of SA in rice. Subsequently, Xoo inoculation experiments were conducted in the field at the tillering stage on wild-type and osd2 mutants. Disease incidence was statistically analyzed 14 days after inoculation. The results showed that the lesion length of the osd2 mutant was significantly increased compared to the wild type, with increases of 60.3%, 43.7%, and 41.7% in three independent lines compared to the wild type, respectively. Figure 1 The results above indicate that OSD2 is involved in the synthesis of SA in rice, and the deletion of the OSD2 gene leads to a significant decrease in the SA content of rice, making it more susceptible to pathogens.

[0095] Example 2: OSD2 gene expression and induction of expression

[0096] The transcriptional expression pattern and regulation of genes are crucial to their biological functions. To investigate the transcriptional expression pattern of the OSD2 gene, qRT-PCR was used to detect the transcriptional levels in the roots, stems, leaves, and panicles of wild-type ZH11 rice at the heading stage. The results showed that the OSD2 gene was expressed in all tissues of rice. Figure 2 A) Because rice is more sensitive to Xoo after SA is missing ( Figure 1 Therefore, samples were taken from ZH11 within 72 hours after inoculation to detect the OSD2 gene induction by Xoo. The results showed that OSD2 was induced by Xoo 24 hours after inoculation, with the expression level increasing by 10.1 times. Figure 1 (B in the text). The above results indicate that SA plays an important role in the disease resistance of rice.

[0097] Example 3: Subcellular localization of OSD2 protein

[0098] Subcellular localization of genes is often closely related to their biological functions. To study the subcellular localization of the OSD2 protein, an OSD2 protein vector fused to the N-terminus of an GFP protein and promoted by a 35S promoter was constructed. Transformation was performed using rice protoplasts, and the subcellular localization was observed. The results showed that the GFP-OSD2 fusion protein co-localized with the peroxisome marker protein mCHerry-AIM1, indicating that OSD2 is localized in the peroxisome (…). Figure 3 ).

[0099] Example 4: OSD2 overexpression plants increased rice resistance to Xoo.

[0100] Overexpression of the OSD2 gene was performed using the ubiquitin promoter (Ubi) in the Zhonghua 11 background. Three overexpression lines were screened, with expression levels 2.4, 5.9, and 11.6 times that of Zhonghua 11, respectively. Figure 4 (A) Xoo inoculation experiments were conducted on the tillering stage, and the disease incidence was observed and statistically analyzed 14 days later. The results showed that, compared with the wild type, the lesion length of the three overexpression lines was reduced by 32.1%, 34.8%, and 31.2%, respectively. Figure 4 (B and C in the figure) indicates that overexpression of OSD2 can increase the resistance of rice to Xoo.

[0101] Based on Examples 1-4 above, it is shown that OSD2 is a gene involved in the biosynthesis of PAL-dependent SA, which is expressed in various plant tissues. Its expression level is induced by Xoo and it is located in the peroxisome. Overexpression of the OSD2 gene can increase the resistance of rice to Xoo.

[0102] The genes SEQ ID NO: 1 and SEQ ID NO: 2 have no introns, therefore their genomes are identical to CDs.

[0103] SEQ ID NO: 1:

[0104]

[0105] SEQ ID NO:2:

[0106]

[0107] SEQ ID NO: 3:

[0108] MAGSTAAALKFTVRRKPAELVAPAGPTPRELKKLSDIDDQDGLRFHIPVIQFYRRSAAMGGRDPAPVIRAAVARALVSYYPFAGRLRELEGRKLAVDCTGEGVLFIEADADVRLEHFGG ALQPPFPCLEELVFDVPGSSEVLGSPLLLFQVTRLACGGFILAVRLHHTMADAQGLVQFLGAVAEMARGGAAAAPSVAPVWGREMLEARSPPRPAFAHREYDEVPDTKGTIIPLDDMAH RSFFFGAREVAAVRSHLAPGIRERATTFEVLTGCLWRCRTAALAPDDDEVMRMICIVNARGGGKSGGGAGMIPEGYYGNAFAFPVAVATAGELRARPLGYAVELVRAAKGEVSVEYMRS VADLMVQRGRPHFTVVRAYLVSDVTKAGFGDLDFGWGKPAYGGPAKGGVGAIPGVASFLIPFKNAKGEDGIVVPMCLPGPAMDKFVEEMGKLMKPAAAATAATRQQPADMFAMIKSAL*

[0109] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions, and improvements made by those skilled in the art within the scope of the technology disclosed in the present invention, and within the spirit and principles of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for regulating the resistance of rice leaves to bacterial blight bacteria, characterized in that, Knocking out the OSD2 gene leads to a decrease in the saline acidity (SA) of rice leaves, making them susceptible to bacterial blight (Xanthomonas oryzae pv. oryzae). Overexpression of the OSD2 gene increases the resistance of rice to bacterial blight. The nucleotide sequence of the OSD2 gene is shown in SEQ ID NO: 1.

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