Application of OsbZIP34 gene in enhancing low-temperature stress resistance of rice in seedling stage

Abstract

The invention relates to application of a rice OsbZIP34 gene in enhancing low-temperature stress resistance in a rice seedling stage, and belongs to the technical field of plant genetic engineering. Specifically, the rice OsbZIP34 gene participating in positive regulation and control of low-temperature stress resistance is obtained through cloning, an overexpression strain and a knockout line of the rice OsbZIP34 gene are constructed, and it is found that compared with a wild plant, the overexpression plant of the rice OsbZIP34 gene shows the characteristic of higher low-temperature resistance, the survival rate is remarkably increased, and the knockout line shows the opposite phenotype. In conclusion, the invention discloses a new function of the rice OsbZIP34 gene, and provides a new target and a new resource for genetic breeding for improving the low-temperature stress resistance of rice.

Description

Technical Field

[0001] This invention relates to the field of plant genetic engineering technology, and more particularly to... OsbZIP34 Application of genes in enhancing rice seedling resistance to low temperature stress. Background Technology

[0002] Rice, a key global food crop, is extremely sensitive to low-temperature stress. Severe cold damage significantly hinders seed germination and seedling growth, leading to substantial yield losses. Statistics show that cold stress causes billions of US dollars in rice yield losses globally each year (Lou et al., 2007), and 3 to 5 million tons in China (Zhu et al., 2015; Sun et al., 2022). Therefore, improving the cold resistance of rice seedlings is of urgent practical significance for mitigating cold damage losses and promoting direct seeding cultivation technology. To address this challenge, researchers have explored various strategies. Among them, pre-treatment with chemicals (such as melatonin) to enhance seed vigor can effectively alleviate cold stress and promote germination and seedling formation (Li et al., 2021). In addition, foliar spraying with optimized chemical formulations is another effective approach (Sun et al., 2020; Back, 2021), but its high cost limits large-scale application.

[0003] At the genetic improvement level, introducing cold-resistant genes through genetic engineering is a core strategy for mitigating the damage caused by low temperatures. Currently, several genes related to cold resistance in rice have been identified, such as... OsCTB4a , OsMAPK3 , OsCBL7 and OsSAPK6 These genes regulate downstream target genes through phosphorylation, participating in the cold adaptation process of rice (Zhang et al., 2017; Li et al., 2022; Jia et al., 2022; Guo et al., 2022; Lou et al., 2022). However, cold resistance in rice is a complex quantitative trait controlled by multiple genes (Chen et al., 2018; Zu et al., 2023; Ding et al., 2023), and the number of cloned and functionally validated cold resistance genes remains limited. Therefore, discovering new cold resistance genes and elucidating their molecular mechanisms is crucial for improving the cold resistance breeding system.

[0004] In the transcriptional regulatory network, DREB1 (dehydration response element binding factor 1) is a key hub in response to low temperature. Low temperature signals activate through multiple pathways. DREB1The expression of [a specific gene] regulates downstream cold-regulatory genes (Ding et al., 2020; Zhang et al., 2022; Li et al., 2022). In rice, OsDREB1 Genes have been shown to respond to cold induction and enhance plant cold resistance (Dubouzet et al., 2003; Ito et al., 2006). Besides DREB1, other transcription factors also constitute a complex regulatory network. For example, rice bZIP73, by coordinating the balance of ABA and ROS, regulates cold resistance in conjunction with bZIP71 (Liu et al., 2018). Recent studies have also revealed several regulatory modules centered around DREB1, such as OsWRKY63-OsWRKY76-OsDREB1B and OsNAC5-OsABI5 (Zhang et al., 2022; Li et al., 2023). Nevertheless, the specific mechanisms by which transcription factor-mediated signaling pathways promote cold resistance still require in-depth analysis, and screening and identifying new cold resistance-related transcription factors is a current research focus.

[0005] The bZIP transcription factor family contains a highly conserved bZIP domain, consisting of a DNA-binding basic region and a dimer-forming leucine zipper region (Wang et al., 2008). Proteins in this family typically recognize cis-acting elements such as ABRE (ABA-responsive elements) and regulate downstream gene expression through post-translational modification or dimerization (Schütze et al., 2008). In rice, OsbZIP34 As a member of the bZIP family, its function and mechanism in low-temperature stress response are still unknown and require further investigation. Summary of the Invention

[0006] Based on existing technologies, this invention has discovered a method for overexpressing [the gene] in rice plants. OsbZIP34 Genes can enhance the resistance of rice plants to low-temperature stress during the seedling stage; therefore, in one aspect, this invention provides rice... OsbZIP34 The application of genes in improving the resistance of rice to low-temperature stress. On the other hand, this invention also provides a method for improving the resistance of rice seedlings to low-temperature stress, comprising: overexpressing rice... OsbZIP34 Genes. Furthermore, this invention also provides rice. OsbZIP34 Application of genes in genetic breeding to improve rice seedling resistance to low temperature stress.

[0007] In this regard, the present invention includes, but is not limited to, the following: In one aspect, the present invention provides rice OsbZIP34The application of the gene in genetic breeding for improving rice seedling resistance to low temperature stress and / or rice variety improvement, i.e., overexpression of the aforementioned rice OsbZIP34 Gene.

[0008] In another aspect, the present invention provides a rice-containing... OsbZIP34 The application of gene recombinant vectors in genetic breeding for improving rice seedling resistance to low-temperature stress and / or rice variety improvement, i.e., overexpression of the aforementioned rice... OsbZIP;34 Gene.

[0009] In one aspect, the recombinant vector of the present invention comprises an overexpression promoter Ubi.

[0010] Preferably, the recombinant vector of the present invention is a recombinant pCAMBIA1301 vector containing the overexpression promoter Ubi.

[0011] In one aspect, the present invention provides the application of genetically engineered bacteria in improving rice seedling resistance to low-temperature stress and / or in genetic breeding for rice variety improvement, wherein the genetically engineered bacteria contains rice OsbZIP34 Gene recombinant vector, overexpressing the rice OsbZIP34 Genes. Preferably, the genetically engineered bacteria of the present invention are genetically engineered Escherichia coli or Agrobacterium.

[0012] In one aspect, the genetically engineered bacteria of the present invention are genetically engineered Agrobacterium.

[0013] In one aspect, the genetically engineered bacterium described in this invention is a genetically engineered Agrobacterium GV3101.

[0014] In another aspect, the present invention provides a method for genetic breeding to improve the resistance of rice seedlings to low-temperature stress and / or to improve rice varieties, the method comprising: overexpressing rice OsbZIP34 Gene.

[0015] In one aspect, the rice variety improvement described in this invention aims to enhance the resistance of rice seedlings to low-temperature stress.

[0016] In another aspect, the present invention provides a method for obtaining rice plants with improved traits, comprising the following processing steps: (1) Infecting rice callus tissue with genetically engineered bacteria; and (2) Infected rice callus tissue was cultured into rice plants; The genetically engineered bacteria contain rice OsbZIP34 Gene recombinant vector, overexpressing the rice OsbZIP34 Gene.

[0017] In another aspect, the present invention provides the application of rice OsbZIP34 protein in improving rice seedling resistance to low temperature stress and / or in genetic breeding for rice variety improvement.

[0018] In one aspect, the amino acid sequence of the rice OsbZIP34 protein of the present invention is shown in SEQ ID NO: 3.

[0019] In one aspect, the improved trait described in this invention is enhanced resistance to low-temperature stress during the rice seedling stage.

[0020] In one aspect, the present invention provides for constructing overexpression OsbZIP34 The method / steps for gene vectors include: [the following steps are described in the original text, which are not directly related to the gene vector method / steps described in the original text.] OsbZIP34 The nucleotide sequence of the gene coding region was ligated into the vector pCAMBIA1301, with the primers designed as follows: OsbZIP34 -ox-F: 5'-ACTAGGGTCTCGCACCATGGCATCCTCGGC-3'; (SEQ ID NO: 4) OsbZIP34 -ox-R: 5'-ACTAGGGTCTCTCGCCCCCTTTGCTGTCATCAT-3'; (SEQ ID NO: 5).

[0021] In one aspect, the present invention also provides for constructing overexpression OsbZIP34 The method / steps for genetically engineered bacteria include: overexpressing the strain described in this invention... OsbZIP34 The gene vector was transformed into the Agrobacterium strain.

[0022] In one aspect, the present invention also provides for obtaining overexpression OsbZIP34 The method / steps for producing gene-modified rice plants include: using the overexpression method described in this invention. OsbZIP34 Genetically engineered bacteria infect rice callus tissue, which is then differentiated and rooted.

[0023] In one aspect, the rice of the present invention OsbZIP34 The gene encodes the amino acid sequence shown in SEQ ID NO: 3.

[0024] In one aspect, the rice of the present invention OsbZIP34 The nucleotide sequence of the gene coding region is shown in SEQ ID NO: 1.

[0025] In one aspect, the rice described in this invention is a japonica rice variety, Nipponbare (Japanese white rice). Oryza.Sativa L.spp. japonica , var. Nipponbare ).

[0026] Preferably, the improved low-temperature stress resistance of rice described in this invention is manifested in that, compared to control ordinary rice plants, OsbZIP34 Rice seedlings with overexpressed genes showed significantly higher survival rates and lower ion permeability after low-temperature treatment during the seedling stage.

[0027] In one aspect, the low temperature described in this invention is below 12°C. In another aspect, the low temperature described in this invention is below 11°C, below 10°C, below 9°C, below 8°C, below 7°C, below 6°C, below 5°C, or below 4°C. In one aspect, the low temperature described in this invention is 4°C.

[0028] In one aspect, the present invention will OsbZIP34 Genes were introduced as the target genes into the japonica rice variety Nipponbare ( Oryza.Sativa L. spp. japonica , var. Nipponbare (obtained from) OsbZIP34 The T0 generation overexpressing plants of the gene were continuously self-crossed to obtain homozygous high-expressing T2 generation lines, which were named […]. OsbZIP34-ox1 and OsbZIP34-ox2 .

[0029] In one aspect, this invention utilizes CRISPR / Cas9 gene editing technology in the japonica rice variety Nipponbare (… Oryza.Sativa L. spp. japonica , var. Nipponbare ) OsbZIP34 Genes were de-functionalized to prepare... OsbZIP34 Plants with missing gene function osbzip34-1 and osbzip34-2 .

[0030] In one aspect, in this invention, OsbZIP34 The nucleotide sequence of the protein-coding region of the gene is shown in SEQ ID NO:1. OsbZIP34 The full-length sequence of the gene is shown in SEQ ID NO: 2. OsbZIP34 The gene encodes a protein belonging to the bZIP family of transcription factors, consisting of 329 amino acids. Its amino acid sequence is shown in SEQ ID NO: 3, specifically: SEQ ID NO: 1:ATGGCATCCTCGGCCGCCTCCCCGTCCTCGTCCAAGTCCGACGACGAGCGGCGGCAGGACGGGGACAGGGACACCCGGGACCCAGCGGCATCATCGTCGGCGGCCGCGGCCGCGGCGCAGACGCACGCGGAGTGGGCGGCGTCGATGCAGGCGTACTACGCGGCCGCCGCCGCCGCCGCCGGTGGCCACCCTTACGCCTGGCCCCCGCCGCAGAGCGTGCCTCCCATGGCTGGTTGCGCCGTGCCATCTGCGGCGGCGGAAGGGAAGAGCAAGAGGAAGACTTCCGGTGGCCCCTCCGGCGAGGACTCCTCCGGGAGTGGCGACGGTGGGAGCGAGGATTCATCGGAGAGGAGAGATGATGCTGACGAGAAGGGTTTGTCACCTGCAAAGTGGAGAAAATTGGGTCATCCAGACATAGAAGGTGAAACATCTCAGGCTGCTGCCATGTCTGAGCAGAATCCTGTGAAGGCGGCACCGAATTTGAATATCGGGATGGATATTTGGAGCAATTCTACAATGGCCGCCATGCCCTCAGGACAGGTGGAGGTGAATGCTGGGACGCATTTGCGGCGAGACAAGGCTTTGTCTCAGATGGATGAACGAGAACTGAAGAGGGAGAGGAGAAAACAATCTAACAGAGAGTCTGCGAGGAGATCGAGACTACGGAAGCAGGAAAGACCCACATCTTTCTGTCACCCGCCAAACTATCCTGTTACTTTTTATGGCCTGACAAACCAAGAATGTGAGGAGTTGTCCCAGAAGGTAACTGAACTGACAGCCGTAAACAGCACACTCATGACAGAACTCGACAAGCTTAAGAAGGACTGTGAAGACATGGAAGCGGAAAATTCGCAGCTAATGGATGAGATGGTGCAGTCCGAGGGCTCTAGTGTTATAGCCACTTTGAGCGTCAAGATTGACACGTCAAAAGACCGCCATGGTAGCAGTAGCCAGCTAAATAAGCACACAAATGATGACAGCAAAGGGTAG SEQ ID NO: 2: SEQ ID NO:3: MASSAASPSSSKSDDERRQDGDRDTRDPAASSSAAAAAAQTHAEWAASMQAYYAAAAAAAGGHPYAWPPPQSVPPMAGCAVPSAAAEGKSKRKTSGGPSGEDSSGSGDGGSEDSSERRDDADEKGLSPAKWRKLGHPDIEGETSQAAAMSEQNPVKAAPNLNIGM DIWSNSTMAAMPSGQVEVNAGTHLRRDKALSQMDERELKRERRKQSNRESARRSRLRKQERPTSFCHPPNYPVTFYGLTNQECEELSQKVTELTAVNSTLMTELDKLKKDCEDMEAENSQLMDEMVQSEGSSVIATLSVKIDTSKDRHGSSSQLNKHTNDDSKG* Compared with the prior art, the present invention has the following beneficial effects: (1) This invention utilizes overexpression and silencing in common rice plants, specifically Nipponbare. OsbZIP34 The gene was discovered to have a novel use in enhancing rice seedling resistance to low-temperature stress and provides an important genetic resource for breeding rice varieties resistant to low-temperature stress.

[0031] (2) This invention utilizes transgenic technology to provide a method for breeding rice germplasm resistant to low-temperature stress, and obtains rice germplasm resistant to low-temperature stress. OsbZIP34 Homozygous plants with gene overexpression. Attached Figure Description

[0032] Figure 1 Rice in Example 2 OsbZIP34 Gene loss-of-function editing target sites and mutation types (A), and OsbZIP34 In gene overexpression materials OsbZIP34 Schematic diagram of gene expression levels (B).

[0033] Figure 2 Rice in Example 3 OsbZIP34 Plants with missing gene function osbzip34-1 and osbzip34-2、 rice OsbZIP34 Gene overexpression plants OsbZIP34-ox1 and OsbZIP34-ox2 Survival rates of wild-type rice plants (Nipponbare) after low-temperature stress; where A is a photograph of plant growth, scale bar = 5 cm; B and C are bar charts of plant survival rate and ion permeability. Detailed Implementation

[0034] To enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. It should be noted that the following detailed descriptions are exemplary and are only some embodiments of the present invention, not all embodiments.

[0035] Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0036] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. The experimental materials used in the embodiments of this invention are all conventional experimental materials in the art and are commercially available. Experimental methods not specifying detailed conditions are performed according to conventional experimental methods or the operating instructions recommended by the supplier.

[0037] Example 1 Rice OsbZIP34 Gene cloning, construction of gene overexpression engineered bacteria and gene editing engineered bacteria 1. Total RNA extraction from rice Total RNA was extracted from young rice leaves using the Tiangen Plant Total RNA Extraction Kit. The extracted RNA was then reverse transcribed according to the instructions of the Thermo Fisher Reverse Transcription Kit to obtain cDNA, which was stored at -20°C for later use.

[0038] 2. OsbZIP34 Construction of gene overexpression engineered bacteria A Design rice OsbZIP34 Primers for specific amplification of gene coding region sequences are as follows: OsbZIP34 -ox-F: 5'-ACTAGGGTCTCGCACCATGGGAGAGGCTAGGAG-3'; (SEQ ID NO: 4) OsbZIP;34 -ox-R: 5'-ACTAGGGTCTCTCGCCGAAAGCTGAAAATTGGCTTTC-3'; (SEQ ID NO: 5) Using the cDNA (complementary DNA) obtained by reverse transcription of total RNA from the leaves of common rice Nipponbare as a template, it was amplified by polymerase chain reaction (PCR) to obtain... OsbZIP34The nucleotide sequence of the gene exon (SEQ ID NO: 1) was obtained, and the PCR amplified fragment was ligated into the vector pCAMBIA1301 containing the overexpression promoter Ubi. After transformation into competent DH5α Escherichia coli by heat shock at 42°C, the fragment was plated.

[0039] Successfully transformed monoclonal DH5α Escherichia coli colonies were selected, and the culture solution after shaking was sent to a sequencing company for sequencing. The sequencing results showed that the vector contained... OsbZIP34 The nucleotide sequences of the gene exons were obtained, and plasmids were extracted using a plasmid extraction kit from TransGen Biotech. The plasmids were electroporated into GV3101 Agrobacterium competent cells, incubated at 28°C for two days, and then plasmids were picked for PCR verification, yielding the product containing the overexpression vector pCAMBIA1301- OsbZIP34 Agrobacterium engineered strain A.

[0040] 3. OsbZIP34 Construction of gene-edited engineered bacteria B based on OsbZIP34 The CDS sequence of the gene (SEQ ID NO: 1) was used to search for a specific target sequence on the online website (http: / / crispr.hzau.edu.cn / CRISPR / ). The target sequence “CCCGTCCTCGTCCAAGTCCGACGACG (SEQ ID NO: 6)” was selected, and complementary primers were synthesized based on this target sequence. Bsa The restriction endonuclease p-1 restriction endonuclease was used to digest the vector pHun4c12, and the linearized vector was obtained by gel extraction. Subsequently, the fusion fragment was ligated into the linearized pHun4c12 vector (Jiang et al., 2019, Mutation of Inositol 1,3,4-trisphosphate 5 / 6-kinase 6 Impairs Plant Growth and Phytic Acid Synthesis in Rice. Plants 8(5): 114). Enzyme digestion confirmed the correct vector was obtained, and further sequencing confirmed that the target sequence had been incorporated into the vector. The correct vector was named pHun4c12- OsbZIP34 Simultaneously, the thermal shock conversion method was used to convert pHun4c12- OsbZIP34 When introduced into Agrobacterium strain EHA105, the gene-editing vector pHun4c12- is obtained. OsbZIP34 Agrobacterium engineered strain B was used for subsequent genetic transformation.

[0041] Example 2 Rice OsbZIP34 Obtaining homozygous plants with gene overexpression and homozygous plants with gene loss of function 1. Agrobacterium-mediated genetic transformation of rice Using pCAMBIA1301- OsbZIP34 Agrobacterium engineered strain A overexpression vector and strain containing pHun4c12- OsbZIP34 Gene-editing Agrobacterium B was used to infect callus tissue of Nipponbare rice, and after screening on differentiation and rooting media, the corresponding genetically transformed plants were obtained.

[0042] 2. Obtaining homozygous overexpression lines A small amount of leaves and roots from the T0 generation overexpression seedlings were taken, and total RNA was extracted from the plant materials (roots and leaves) using the RNeasy Plant RNA Mini Kit (Qiagen, Hilden, Germany). cDNA was then reverse transcribed using 1 μg of total RNA, oligo-dT18 primers, and the GoScript™ reverse transcription system (Promega). Quantitative real-time polymerase chain reaction (qRT-PCR) was performed using a SYBR Green GoTaq qPCRMaster Mix (Promega, WI, USA). Rice OsACTIN Genes were used as internal controls, and 2 were used. -ΔΔCt Method calculation OsbZIP34 Relative expression level.

[0043] Table 1 Primer list for quantitative real-time polymerase chain reaction (qRT-PCR)

[0044] T0 generation transgenic overexpression plants were self-pollinated to obtain T1 generation seeds. From each T0 generation overexpression plant, 6-9 positive plants were taken and self-pollinated again to produce T2 generation seeds, which were then segregated and analyzed. When all T2 generation seedlings produced from the positive T1 generation plants tested positive, the T1 generation plant was considered a homozygous overexpression plant, and thus the T1 generation was obtained. OsbZIP34 Transgenic pure lines with overexpression of the gene are named OsbZIP34-ox1 and OsbZIP34-ox2 In the two overexpression lines OsbZIP34 See gene expression levels. Figure 1 B; conversely, it is a heterozygous plant.

[0045] 3. Obtaining homozygous plants with gene function loss Young leaves were selected from T0 generation transgenic rice at the 3-4 leaf stage, and genomic DNA was extracted using the CTAB method. Endogenous DNA from the transgenic rice was amplified using the pHun4c12-OsbZIP34 specific primers. OsbZIP34 Gene sequencing was used to verify the PCR products, and the results showed that... OsbZIP34 The gene has undergone a single base deletion, i.e., acquired OsbZIP34 Transgenic pure lines with gene function loss are named osbzip34-1 and osbzip34-2(See) Figure 1 A).

[0046] Example 3 OsbZIP34 Experiments on the resistance of rice to low temperature stress and determination of related gene expression. The rice obtained in Example 2 OsbZIP34 Overexpression of T2 generation homozygous lines: OsbZIP34-ox-1 and OsbZIP34- ox-2 , OsbZIP34 Transgenic pure lines with gene function loss: OsbZIP34-1 and OsbZIP34-2 Compared with ordinary rice plants, low temperature stress was applied.

[0047] 1. Low temperature stress treatment of rice To assess the cold resistance of rice plants, rice seedlings soaked for 10 days were transferred to either 4℃ (experimental group) or 30℃ (control group) for 6 days, followed by a 6-day recovery period at 30℃, and survival rates were analyzed. All treatments included three biological replicates of 30 to 40 plants for survival determination. All tests were conducted in an artificial indoor environment under the following conditions (relative humidity: 65–75%; 16 hours light / 8 hours darkness). Meanwhile, the ion permeability of different strains after low-temperature treatment was detected. The ion leakage test procedure was as follows: Ion leakage was quantified according to Ding et al. (Ding, Y., Li, H., Zhang, X., Xie, Q., Gong, Z., Yang, S. (2015). OST1 kinase modulates freezing tolerance by enhancing ICE1 stability in Arabidopsis. Dev. Cell 32: 278–289). Specifically, approximately 0.1 g of leaf tissue from low-temperature treated seedlings was placed in a 15 mL test tube, and 5 mL of deionized water was added. The sample was gently shaken at 200 rpm for 1 hour at room temperature, and the initial conductivity (A1) was recorded. The sample was then heated in boiling water for 20 minutes, and then incubated again at room temperature for 1 hour, and the final conductivity (A2) was measured. The conductivity of deionized water (A0) was used as the baseline. The ion leakage rate was calculated as: Ion leakage (%) = (A1) / (A2) / (A3) / (A4) / (A5) / (A6) / (A7) / (A8) / (A9) / (A2 ... A0) / (A2) A0).

[0048] Depend on Figure 2 It can be seen that by overexpressing homozygous lines in the T2 generation... OsbZIP34-ox1 and OsbZIP34-ox2 , OsbZIP34 Gene knockout plants osbzip34-1 and osbzip34-2After cold treatment for 6 days and then cultured under normal conditions for 6 days, the survival rate of the 10-day-old plants of the control rice variety Nipponbare was recorded. OsbZIP34 The survival rate of overexpressing rice was significantly higher than that of the control after low-temperature treatment; OsbZIP34 Gene knockout plants osbzip34-1 and osbzip34-2 The survival rate of rice after low-temperature treatment was significantly lower than that of the control group. Meanwhile, OsbZIP34 The ion permeability of the overexpression lines was significantly lower than that of the control, while OsbZIP34 Gene knockout lines are the opposite ( Figure 2 C), indicating overexpression OsbZIP34 Genes contribute to maintaining cell homeostasis under low-temperature stress, a result consistent with the aforementioned survival statistics, jointly demonstrating that... OsbZIP34 The gene can significantly enhance the resistance of rice to low temperature stress.

Claims

1. Rice OsbZIP34 The application of genes in genetic breeding for improving rice seedling resistance to low temperature stress and / or rice variety improvement is characterized by, Overexpression of the rice OsbZIP34 Gene.

2. Contains rice OsbZIP34 The application of gene recombinant vectors in genetic breeding for improving rice seedling resistance to low-temperature stress and / or rice variety improvement is characterized by, Overexpression of the rice OsbZIP34 Gene.

3. The application of a genetically engineered bacterium in improving rice seedling resistance to low-temperature stress and / or in genetic breeding for rice variety improvement, characterized in that... The genetically engineered bacteria contain rice. OsbZIP34 Gene recombinant vector, overexpressing the rice OsbZIP34 Gene.

4. A method for improving rice seedling resistance to low-temperature stress and / or for genetic breeding of rice varieties, characterized in that, The method includes: overexpressing rice OsbZIP34 Gene.

5. A method for obtaining rice plants with improved traits, characterized in that, It includes the following processing steps: (1) Infecting rice callus tissue with genetically engineered bacteria; and (2) Infected rice callus tissue was cultured into rice plants; The genetically engineered bacteria contained rice OsbZIP34 Gene recombinant vector, overexpressing the rice OsbZIP34 Gene.

6. The application according to any one of claims 1-3 or the method according to claim 4 or 5, characterized in that, The rice OsbZIP34 The gene encodes the amino acid sequence shown in SEQ ID NO:

3.

7. The application or method according to claim 6, characterized in that, The rice OsbZIP34 The nucleotide sequence of the gene coding region is shown in SEQ ID NO:

1.

8. Application of rice OsbZIP34 protein in improving rice seedling resistance to low temperature stress and / or in genetic breeding for rice variety improvement.

9. The application according to claim 8, characterized in that, The amino acid sequence of the rice OsbZIP34 protein is shown in SEQ ID NO: 3.

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