Application of rice os-er-ant1 gene

Abstract

The application relates to an application of a rice Os-ER-ANT1 gene. The nucleotide sequence of the gene is shown in SEQ ID No. 1, the encoded amino acid sequence is shown in SEQ ID No. 2, the gene can regulate rice photorespiration amino acid metabolism, can be used for rice amino acid metabolism improvement breeding, and provides a new genetic resource for improving crop quality and yield.

Description

Technical Field

[0001] This invention belongs to the field of molecular biology, and specifically relates to the application of a rice gene Os-ER-ANT1 that regulates amino acid metabolism in improving crop quality and yield. Background Technology

[0002] Rice yield is influenced by a variety of factors, including the number of effective panicles, the number of grains per panicle, and the thousand-grain weight. Meanwhile, the photosynthetic efficiency of rice is also a crucial factor affecting yield; however, photorespiration, a process involved in photosynthesis, is generally considered to consume energy and reduce carbon assimilation efficiency. Nevertheless, photorespiration also plays a protective role in plant adaptation to environmental changes, such as mitigating photoinhibition, rapidly adapting to low CO2 environments, promoting nitrogen assimilation, and coupling amino acid metabolism.

[0003] Photorespiration is a complex metabolic process involving multiple organelles, including chloroplasts, peroxisomes, and mitochondria. Ribose-1,5-bisphosphate carboxylase / oxygenase (Rubisco) is the main enzyme involved. Under CO2 conditions, it catalyzes the carboxylation of ribose-1,5-bisphosphate to produce 3-phosphoglycerate (3-PGA), while under oxygen conditions, it undergoes oxygenation to produce 3-PGA and 2-phosphoglycolic acid (2-PG). The conversion of 2-PGA to 3-PGA in photorespiration is energy-intensive, but it also plays an important physiological role through interactions with glycine and serine produced by photorespiration, coupled with nitrogen assimilation, via amino acid metabolism.

[0004] The endoplasmic reticulum (ER) also plays a crucial role in photorespiratory metabolism, and the ER is home to ATP / ADP transporters ( Us-IS- ANT1 It is believed that it can provide ATP to the ER. In Arabidopsis and rice... Us-ER-ANT1 The deletion mutant exhibits a typical photorespiration phenotype in air, such as inhibited growth or dwarfism, but can grow normally under high CO2 conditions. Furthermore, Arabidopsis thaliana... is-ant1 The mutant exhibits glycine accumulation due to inhibited glycine decarboxylase (GDC), negatively impacting plant development. However, this glycine accumulation is not the sole cause of its dwarfing phenotype. Studies have shown that the endoplasmic reticulum plays a crucial role in photorespiration metabolism, but... Us-ER-ANT1 Its specific physiological functions and its effects on amino acid metabolism are not yet fully understood.

[0005] Key enzymatic steps in the photorespiration pathway are distributed across multiple subcellular organelles, and amino acid metabolism plays a crucial role in the metabolic connections between these organelles. However, Us-ER-ANT1 Its role in linking photosynthesis and amino acid metabolism remains unclear.

[0006] This study investigated the effects of air, elevated CO2 conditions, and the transition from high CO2 to air on rice. is-ant1 A comprehensive analysis of the amino acid metabolism of the mutant revealed ER-ANT1 This invention relates to a potential mechanism by which endoplasmic reticulum ATP / ADP transporters may be involved in amino acid metabolism under photorespiratory conditions, providing new insights into the role of endoplasmic reticulum ATP / ADP transporters in rice photorespiration and amino acid metabolism. The invention also relates to a rice gene regulating amino acid metabolism. Us-ER-ANT1 This provides new gene resources for the genetic improvement of rice amino acid metabolism, and is expected to be used to breed new rice varieties with high photosynthetic efficiency and high yield. Summary of the Invention

[0007] The purpose of this invention is to provide rice genes. Us-ER-ANT1 Applications. Us-ER-ANT1 This gene plays a crucial role in the photorespiratory metabolism of rice, regulating amino acid metabolism by influencing ATP / ADP transport and energy metabolism. Therefore, cloning this gene can not only elucidate the molecular mechanisms regulating amino acid metabolism in rice but also provide new gene resources for the genetic improvement of crop amino acid metabolism.

[0008] This invention is achieved through the following technical solution:

[0009] This invention provides the application of the Os-ER-ANT1 gene in regulating amino acid metabolism during photorespiration in rice. The nucleotide and amino acid sequences of this gene are as follows:

[0010] A. Nucleotide sequence: The DNA sequence shown in SEQ ID NO: 1;

[0011] B. Amino acid sequence: The amino acid sequence shown in SEQ ID NO: 2.

[0012] Containing the above ER-ANT1 The application of gene knockout vectors, overexpression vectors, transgenic strains, and recombinant bacteria also falls within the scope of protection of this invention.

[0013] This invention utilizes targeted knockout of rice Us-ER-ANT1 Mutant material of gene (LOC_Os11g43960) Us-ER-ANT1 This mutant exhibits stunted growth and abnormal amino acid metabolism in air, particularly abnormal accumulation of glycine, glutamic acid, and alanine. Experimental results show that under high CO2 conditions, the mutant's amino acid metabolism partially recovers, while in air, it exhibits drastic changes in amino acid content, especially the accumulation of glycine. Us-ER-ANT1 The gene is an adenosine nucleotide transporter located in the endoplasmic reticulum, indicating that... Us-ER-ANT1 It is a key regulator of amino acid metabolism. Currently, there is no information regarding... Us-ER-ANT1Studies on amino acid metabolism in rice photorespiration have been reported, therefore the target gene of this invention... Us-ER-ANT1 It is a new gene that regulates amino acid metabolism in rice.

[0014] The above-mentioned rice Us-ER-ANT1 The application of the gene also includes its application in rice amino acid metabolism improvement breeding. The methods for improving rice amino acid metabolism include genetic engineering breeding and hybridization breeding. The method involves genetic engineering breeding, where the gene contains… Us-ER-ANT1 Gene knockout vectors are introduced into plant cells, knocking out genes in the plant. Us-IS- ANT1 Materials obtained through genetic modification that alter amino acid metabolism, such as those with knockout genes in rice. Us-ER-ANT1 Genes can be used to breed rice varieties with abnormal photorespiration and amino acid metabolism. Hybrid breeding: Using rice materials with the aforementioned amino acid metabolism mutations as parents, the amino acid metabolism of rice is improved through hybridization and selection. is-ant1 Mutant rice was hybridized with rice varieties intended for improvement to achieve targeted improvement of rice amino acid metabolism.

[0015] The present invention has the following advantages and effects:

[0016] 1. ER-ANT1 Gene deletion, manifested in rice Us-ER-ANT1 Gene inactivation and phenotypic analysis showed that the mutant exhibited slow growth and abnormal amino acid metabolism. This provides new gene resources for the genetic improvement of rice amino acid metabolism and offers new target genes for molecular design breeding of rice.

[0017] 2. There is currently no information regarding... Us-ER-ANT1 Research on the regulatory function of genes in rice amino acid metabolism. Therefore, the target gene of this invention... Us-ER-ANT1 This is a novel gene regulating amino acid metabolism in rice. It holds significant theoretical importance in elucidating the mechanisms of amino acid metabolism and provides new evidence for broadening our understanding of the existing molecular mechanisms regulating amino acid metabolism in rice. Attached Figure Description

[0018] Figure 1 For Example 1, wild type and is-ant1 The phenotype of the mutant and its leaf amino acid content, Figures A and B are wild-type WT and is-ant1 The phenotypes of mutants after 15 and 30 days of culture are shown in Figure C, which shows the amino acid content of leaves from plants harvested at 6 PM after 12 days of growth under natural conditions. Figure D shows the amino acid content of plants grown under high CO2 conditions. is-ant1 The amino acid content of leaves of wild-type plants and medium-sized plants was compared, with the WT of wild-type plants being 11.

[0019] Figure 2Example 2: Wild type and... is-ant1 Comparison of free amino acid content in mutants; Figure A shows the results of transfer from high CO2 treatment for 12 hours to natural conditions for wild-type and... is-ant1 The changes in total amino acid content of the plant are shown in Figure B, which shows the proportion of each amino acid, and Figure C, which shows the changes in the content of glycine, valine, isoleucine, leucine, phenylalanine, and histidine.

[0020] Figure 3 Example 2 Wild type and is-ant1 Changes in glutamate content in mutants; Figure A shows changes in glutamate and glutamine content; Figure B shows Fd-GOGAT activity and mRNA levels; Figure C shows changes in control ZH8015 and... es7 A comparison of whole plants of the mutant grown in paddy fields; Figure D shows the control ZH8015 and... es7 Comparison of leaf phenotypes of mutants grown in paddy fields; Figure E shows the leaf phenotypes of 3-week-old ZH8015 and... es7 Amino acid content of leaves in ambient air.

[0021] Figure 4 Example 2: Correlation between alanine content, AGAT1 activity, and AGAT1 mRNA levels; Figure A shows the phylogenetic tree of eight transaminases in Arabidopsis and rice that are presumed to be related to photorespiration; Figure B shows the wild-type and... is-ant1 Changes in alanine content in plants, as shown in Figure C. Us-AGAT1 Gene expression levels, Figure D shows wild-type and... is-ant1 After the mutant was transferred from a high CO2 environment to the air for 7 days, the alanine content... AGAT1 Comparison of transcriptional levels and AGAT1 enzyme activity.

[0022] Figure 5 For Example 2, under photorespiration conditions, is-ant1 The phosphoserine pathway that induces serine synthesis; Figure A shows the wild-type and... is-ant1 The changes in serine content in mutant leaves. Figure B shows the gene expression analysis of genes encoding phosphoserine phosphatase (PSP), phosphoserine transaminase (PSAT), and 3PGA dehydrogenase (3-PGDH) under high CO2 and natural conditions.

[0023] Figure 6 Example 3: Wild-type and is-ant1 plant NAD + And NADH content; plants were grown under high CO2 conditions for 2 weeks and then transferred to natural conditions, and leaf materials were harvested at 0, 1, 8 and 12 hours.

[0024] Figure 7 Example 4 Wild type and is-ant1The mutant's phenotype was observed when it grew in a high CO2 environment and then transferred to ambient air; Figure A shows the wild type and... is-ant1 The mutant was grown under high CO2 conditions. Figure B shows the phenotype of the plant after 2 weeks of growth under natural conditions. Figure C shows the photorespiration rate determined using the Rubisco enzyme kinetic estimation method. Figure D shows the photorespiration rate at a photon flux density of 1100 μmol / m³. 2 s 1 The temperature was 28℃ and the CO2 concentration was approximately 400 μmol. 1 The net photosynthetic rate was determined under the specified conditions. Detailed Implementation

[0025] The present invention will be further illustrated below with reference to specific embodiments, but these embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field. Unless otherwise specified, the reagents and materials used in the following embodiments are commercially available.

[0026] The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and substance of the invention are within the protection scope of the present invention.

[0027] In this example, the rice variety is wild-type japonica rice Zhonghua 11 (a publicly available rice variety, commercially available), the pCubi1390 vector is a commonly used gene expression vector, and Escherichia coli DH5α and Agrobacterium tumefaciens EHA105 are commonly used strains, which are preserved in most molecular biology laboratories.

[0028] The main reagents used in the examples were: restriction endonucleases, Taq polymerase, T4 ligase, Pyrobest Taq polymerase, KOD, etc., purchased from biotechnology companies such as TAKARA (Dalian), Promega, NEB, and ABM; plasmid miniprep kits and agarose gel extraction kits were purchased from Shanghai Jierui Biotechnology Co., Ltd.; culture media, agar powder, agarose, ampicillin (Amp), kanamycin (Kan), rifampin (Rif), etc., were purchased from Sigma; other chemical reagents used in the examples were imported or domestically produced analytical grade. Primers used in the examples were synthesized by Beijing Ruiboxingke Biotechnology Co., Ltd., and sequencing was performed by Beijing Qingke Biotechnology Co., Ltd.

[0029] The testing methods involved are as follows:

[0030] Gas exchange measurement

[0031] Gas exchange measurements were performed using a GFS-3000 system. High CO2 conditions (i.e., non-photorespiration conditions) were employed in a sealed artificial climate chamber with continuous dry ice supply. The temperature range was 28–30°C, relative humidity range was 70–90%, and photon flux density ranged from ~300 μmol / m³. 2 s 1 Two weeks after growth, gas exchange was measured in the youngest, fully expanded leaves of each three-leaf seedling. Under normal growth conditions (i.e., photorespiration conditions, temperature setting: 28℃, relative humidity: 70%, photon flux density: 1100 μmol / m³), the gas exchange was measured. 2 s 1 CO2: 400 μmol 1 ), measuring the net CO2 assimilation rate of leaves (A net ) and intercellular CO2 concentration (C i Mitochondrial respiration rate under light (R) d ) by nighttime dark breathing rate (R n The latter was replaced by a method for measuring photorespiration in the dark. The photorespiration rate (P0) was calculated using the Rubisco enzyme kinetic estimation method. R ),

[0032] ,

[0033] in Γ * This is the CO2 compensation point for the chloroplast carboxylation site. C c The CO2 concentration in chloroplasts is calculated using the following formula:

[0034] ,

[0035] Among them, mesophyll electrical conductivity (g) m = 0.45 mol m 2 s 1 ).

[0036] Leaf Fd-GOGAT activity assay

[0037] Soluble total protein was extracted from rice leaves at 0-4°C using HEPES buffer (50 mmol / L). 1 (pH 7.5), add 0.1% β-mercaptoethanol, 1 mM ethylenediaminetetraacetic acid (EDTA) and 5 mmol L 1The MgCl2-Fd-GOGAT activity assay used reduced methyl ozonafil as an electron donor. The reaction mixture consisted of 50 mmol / L... 1 HEPES (pH 7.5), 10 mmol L 1 Glutamine, 10 mmol L 1 2-Ketoglutarate, 15 mmol L 1 The mixture consisted of shellac methyl ester and crude extract with a final volume of 0.5 mL. After pre-incubation at 30°C for 5 min, a reducing agent solution (12 mmol / L) was added. 1 Na2S2O4, 13 mmol L 1 The reaction was initiated with NaHCO3. After incubation at 30°C for 20 min, 0.5 mL (6%) salicylsulfonic acid was added, and the reaction was terminated by vigorous shaking. Fd-GOGAT activity was calculated by measuring the increase in glutamic acid using an amino acid analyzer. A blank control (without glutamine) was added in each experiment.

[0038] Leaf AGAT activity assay

[0039] The assay of AGAT activity is similar to that of GGAT activity, the only difference being that the amino donor (alanine) and pyruvate dehydrogenase are modified as coupling enzymes. The AGAT assay includes 10 mmol L... 1 L-alanine, 10 mmol L 1 Glyoxylate and 100 μL crude enzyme (for enzyme assay) or 100 μL 50 mmol / L 1 PBS (potassium salt, pH 7.4) was used as a control at 100 mmol / L. 1 Add 0.5 U pyruvate dehydrogenase and 0.25 mmol L to HEPES-NaOH (pH 8.0). 1 CoA and 0.2 mmol L 1 NADH, final volume 0.5 mL.

[0040] Amino acid content determination

[0041] Rice leaves were harvested using surgical scissors, rapidly frozen in liquid nitrogen, and then ground into powder using a pestle in a mortar. 0.2 g of the powder sample was extracted on ice for 40 min with 2 mL of 6% (w / v) 5-salicylsulfonic acid. Total free amino acids were determined using an amino acid analyzer equipped with a column, eluted for 148 min at 570 and 440 nm using a pH-dependent lithium buffer gradient elution program. For each amino acid, the mean of three replicates was calculated, with each replicate consisting of 10–12 plants.

[0042] NADH / NAD + Ratio determination

[0043] NAD + NADH was determined using a quantitative kit from Nanjing Jiancheng. NAD+ was determined using the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide) reduction method. + And NADH content. Rice leaves (~200 mg) were first grown in a high CO2 environment, then grown in ambient air for 0, 1, 8, and 12 hours. The tissues were first ground in an ice bath, extracted with 2 mL of NADH / NAD+ alkaline / acidic extraction buffer, and boiled for 5 minutes. After cooling in ice, the samples were subjected to 10,000... g Centrifuge at 4°C for 10 min. Then transfer the supernatant to a new centrifuge tube, mix with the corresponding volume of extract, and perform a neutralization reaction. Incubate the mixture at 4°C for 10,000 minutes. g Centrifuge for 10 min and collect the supernatant for analysis. Mix 200 μL of the sample supernatant with reagents A, B, and C, and incubate at room temperature for 25 min. The sample is then analyzed at 20,000... g Centrifuge for 5 min, mix the precipitate with reagent D, and measure the absorbance of each tube at 570 nm. NAD + The formula for calculating NADH content is as follows:

[0044]

[0045] W represents the fresh weight of the sample to be tested, and V represents the volume of the extract (2 ml).

[0046] In the following examples, the method described in the example of patent CN104673804B was first followed to obtain the Os-ER-ANT1 gene (LOC_Os11g43960) from Zhonghua 11 rice by targeted knockout of the rice Os-ER-ANT1 gene. is-ant1 Mutants. These were observed in rice under conditions of air, elevated CO2, and transition from high CO2 to air. is-ant1A comprehensive analysis of the phenotype and amino acid metabolism of the mutant and conventional wild-type WT plants (Zhonghua 11) was conducted to reveal... ER-ANT1 Potential mechanisms that may be involved in amino acid metabolism under photorespiration conditions.

[0047] Example 1 Wild type and is-ant1 The phenotype of the mutant and its leaf amino acid content

[0048] Wild-type WT and Us-are-ant1 Photographs were taken of the mutant plants after 15 and 30 days of natural growth, as shown. Figure 1 As shown, Figure A represents wild-type WT and is-ant1 Figure B shows the phenotype of the mutant after 15 days of culture, while Figure B shows the phenotype after 30 days of culture.

[0049] Grown for 12 days under natural conditions, and under high CO2 conditions (i.e., non-photorespiration conditions), in a sealed artificial climate chamber with continuous dry ice supply, at a temperature range of 28-30°C, a relative humidity range of 70-90%, and a photon flux density range of ~300 μmol / m³. 2 s 1 Wild-type WT and Us-are-ant1 The amino acid content of the leaves of the mutant plants was analyzed. The results are as follows: Figure 1 As shown in C and D, it can be seen that under photorespiration conditions (natural conditions, ~0.4% CO2), is-ant1 The mutant exhibits slow growth and abnormal amino acid metabolism; however, under non-photorespiration conditions (high CO2 conditions, 1.5% CO2), the content of most amino acids in the mutant is not different from that in the wild type.

[0050] Example 2: Wild-type and is-ant1 Amino acid content analysis of mutants.

[0051] To better understand is-ant1 Metabolic changes associated with the photorespiration phenotype in mutants were observed under high carbon dioxide conditions (i.e., non-photorespiration conditions) in a closed artificial climate chamber with continuous exposure to dry ice, at a temperature range of 28–30°C, a relative humidity range of 70–90%, and a photon flux density range of ~300 μmol / m³. 2 s 1 Treat for 12 hours, then transfer to natural conditions (~0.4% CO2). Detect wild-type and... is-ant1The changes in leaf amino acids of plants at different times after being transferred to natural conditions were studied. Meanwhile, the ZH8015 and es7 mutants grown in paddy fields were used as controls. Figure 3 C and D show the comparison of the whole plant and leaf phenotypes of the ZH8015 and es7 mutants grown in paddy fields, respectively.

[0052] Figure 2 The wild type and the wild type at different times after being transferred to natural conditions are given. is-ant1 The free amino acid content of plant leaves is shown in Figure A, which shows the change in total amino acid content. Figure B shows the proportion of each amino acid. Figure C shows the changes in the content of glycine, valine, isoleucine, leucine, phenylalanine, and histidine.

[0053] Figure 3 A represents the wild-type and [other types] at different times after being transferred to natural conditions. is-ant1 Changes in glutamate and glutamine content in plant leaves. Figure B shows Fd-GOGAT activity and mRNA levels. Figure E shows the amino acid content in the leaves of 3-week-old control groups ZH8015 and es7 in ambient air.

[0054] Figure 4 The correlation between alanine content, AGAT1 activity, and AGAT1 mRNA levels is shown. Figure A shows the phylogenetic tree of eight transaminases in Arabidopsis thaliana and rice that are presumed to be related to photorespiration. Figure B shows the wild-type and... is-ant1 Changes in alanine content in plants. Figure C shows... Us-AGAT1 Gene expression levels. Figure D shows wild-type and... is-ant1 After the mutant was transferred from a high CO2 environment to the air for 7 days, the alanine content... AGAT1 Comparison of transcriptional levels and AGAT1 enzyme activity.

[0055] Figure 5 Under photorespiration conditions, is-ant1 The phosphoserine pathway that induces serine synthesis. Figure A shows the wild-type and... is-ant1 Changes in serine content in mutant leaves. Figure B shows the gene expression analysis of genes encoding phosphoserine phosphatase (PSP), phosphoserine transaminase (PSAT), and 3PGA dehydrogenase (3-PGDH) under high CO2 and natural conditions.

[0056] Combination Figure 2-5 It can be seen that wild type and is-ant1 The total free amino acid content at 0 h was similar. However, after being transferred to natural conditions, the wild-type and... is-ant1The amino acid content of the leaves underwent various changes. Specifically, the contents of glutamate (Glu), glutamine (Gln), and alanine (Ala) decreased; serine levels initially decreased, but accumulated after one week. Alanine:glyoxylate aminotransferase (AGAT) is a key enzyme in the glycine synthesis pathway, catalyzing the conversion of alanine to glycine. AGAT1 is an endoplasmic reticulum localized protein... is-ant1 The mutant exhibits higher activity, which may be due to the decreased alanine level and the large accumulation of glycine (Gly).

[0057] Example 3 Wild-type and is-ant1 plant NAD + and NADH content.

[0058] Combine wild type and is-ant1 Plants were grown under high CO2 conditions for two weeks and then transferred to natural conditions. Leaf materials were harvested at 0, 1, 8, and 12 hours to test NAD. + And NADH content, the results are as follows Figure 6 As shown.

[0059] Glycine from photorespiration is the main source of NADH in leaves. Determination of wild-type and... is-ant1 NAD during CO2 conversion + The abundance of NADH and the results showed that under high CO2 conditions, wild-type and is-ant1 There is no difference in NADH between them, while is-ant1 mutant NAD + Lower than the wild type. After being transferred to natural conditions, the wild type's NADH level increased, while... is-ant1 Higher levels of NAD in mutants; higher levels in wild-type. + The content increases, while is-ant1 NAD in mutants + Almost no change ( Figure 6 This indicates that in er- ant1 NAD in mutants + Metabolic patterns have changed. The above results indicate that... Us-ER-ANT1 Missing NAD + Metabolism leads to NADH / NAD + The ratio increased.

[0060] Example 4 Wild type and is-ant1 The mutant's phenotype was observed when it grew in a high CO2 environment and then transferred to ambient air.

[0061] Combine wild type and is-ant1 After growing under high CO2 conditions for 2 weeks, the plants were transferred to natural conditions and their phenotypes were assessed, including the photorespiration rate and net photosynthetic rate.

[0062] Figure 7 Figure A represents the wild type and is-ant1 The mutants were grown under high CO2 conditions. Figure B shows the phenotype of plants transferred to natural conditions for 2 weeks. Figure C shows the phenotypes of wild-type and wild-type plants after 2 weeks of growth under natural conditions, as determined by Rubisco enzyme kinetic estimation. is-ant1 Photorespiration rate of the plant. Figure D shows the photorespiration rate at a photon flux density of 1100 μmol / m³. 2 s 1 Temperature is 28℃, CO 2 The concentration is approximately 400 μmol. 1 Determining wild-type and under the conditions is-ant1 Net photosynthetic rate of plants after being transferred to natural conditions for 2 weeks.

[0063] It can be seen that under high CO2 conditions, is-ant1 The mutants grew similarly to the wild-type plants; however, after approximately two weeks of exposure to ambient air, is-ant1 The mutant withered and nearly died. Under natural growth conditions, is-ant1 The mutant's photorespiration and net photosynthetic rate were suppressed.

[0064] In conclusion, this invention concludes that: rice is-ant1 The mutant exhibits slow growth and abnormal amino acid metabolism, but under high carbon dioxide conditions (1.5% CO2), the content of most amino acids in the mutant is not significantly different from that in the wild type. Us-IS- ANT1 Deficiency causes NADH / NAD + An increased ratio leads to decreased GDC activity, hindering glycine breakdown and thus reducing downstream metabolites such as serine. This indicates that... Us-ER-ANT1 It is a housekeeping gene with important functions, playing a crucial role in regulating rice amino acid metabolism and can be used in rice amino acid metabolism improvement breeding.

[0065] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

[0066] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0067] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The application of the rice Os-ER-ANT1 gene in inhibiting amino acid metabolism during photorespiration in rice, characterized by: The nucleotide sequence of the Os-ER-ANT1 gene is shown in SEQ ID No. 1, and the encoded amino acid sequence is shown in SEQ ID No. 2; the application is to inhibit the amino acid metabolism of rice by knocking out the expression of the Os-ER-ANT1 gene.

2. Application of the rice Os-ER-ANT1 gene in rice amino acid metabolism improvement breeding, wherein the improvement is to inhibit rice amino acid metabolism, the nucleotide sequence of the Os-ER-ANT1 gene is shown in SEQ ID No. 1, and the encoded amino acid sequence is shown in SEQ ID No. 2; the application is to knock out the Os-ER-ANT1 gene in the plant through genetic engineering.

Images