Hevea brasiliensis ERF transcription factor HbERF105 gene and its application

Abstract

The invention discloses a rubber tree ERF transcription factor HbERF105 gene and an application thereof, and belongs to the technical field of genetic engineering. The invention clones an ERF transcription factor HbERF105 gene in a rubber tree, and its nucleotide sequence is shown in SEQ ID NO.3. It is proved by molecular biology experiments and genetic engineering methods that the gene actively responds to low temperature stress, can effectively improve the cold resistance of transgenic plants, provides gene resources for cold-resistant molecular breeding of rubber trees, and effectively solves the current situation of lack of cold-resistant varieties of rubber trees and low breeding efficiency.

Description

Technical Field

[0001] The invention relates to the technical field of genetic engineering, in particular to a rubber tree ERF transcription factor HbERF105 gene and an application thereof. Background Art

[0002] Rubber trees are typical tropical cash crops, native to the Amazon River Basin in Brazil. The world's recognized suitable planting area is 10°S to 15°N, while some rubber tree planting areas are located between 18°N and 24°N, which are listed as non-traditional rubber planting areas. They are the northernmost rubber planting areas in the world and often suffer from low temperatures below 10°C. When the ambient temperature is lower than the critical cold damage temperature of rubber trees (15°C), cold damage will occur to the tree body, and the lower the temperature, the more serious the cold damage. Under normal circumstances, when the temperature is ≤10°C, the metabolism of young rubber trees will be adversely affected, and photosynthesis will stop; when the temperature is ≤5°C, rubber trees will suffer from varying degrees of cold damage, and a small amount of peeling and gum will flow at the junction of the cut surface and the tree stem, and the treetops and trunks will die, resulting in a decrease in rubber production.

[0003] Low temperature stress has become the main bottleneck restricting the development of the natural rubber industry. At present, the cold-resistant breeding of rubber trees mainly adopts hybrid breeding, and it takes nearly 30 years to cultivate a new variety. The rubber tree breeding cycle is long and the efficiency is low, which can no longer meet the demand for varieties in the natural rubber industry. Therefore, it has become an important part of rubber tree breeding to discover cold-resistant genes, analyze their molecular mechanisms of responding to low temperature stress, improve the cold resistance of rubber tree varieties through molecular breeding technologies such as genetic engineering, enhance the adaptability of varieties, and speed up the breeding of rubber tree varieties. Summary of the invention

[0004] The object of the present invention is to provide a rubber tree ERF transcription factor HbERF105 gene and its application, to solve the problems existing in the above-mentioned prior art. The present invention clones a ERF transcription factor HbERF105 gene in rubber tree, and proves through molecular biology experiments and genetic engineering means that the gene actively responds to low temperature stress, can effectively improve the cold resistance of transgenic plants, provides gene resources for cold-resistant molecular breeding of rubber tree, and effectively solves the current situation of lack of cold-resistant varieties of rubber tree and low breeding efficiency.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The invention provides a rubber tree ERF transcription factor HbERF105 gene, and the nucleotide sequence of the HbERF105 gene is shown as SEQ ID NO.3.

[0007] Preferably, the amino acid sequence of the protein encoded by the HbERF105 gene is shown in SEQ ID NO.4.

[0008] The present invention also provides a recombinant vector, which contains the HbERF105 gene.

[0009] The present invention also provides a recombinant bacterium, wherein the recombinant bacterium comprises the HbERF105 gene or the recombinant vector.

[0010] The present invention also provides the use of the HbERF105 gene and the protein encoded by it, the recombinant vector or the recombinant bacteria in regulating the cold resistance of plants.

[0011] The present invention also provides a method for regulating the cold resistance of plants, comprising the steps of using genetic transformation technology to transfer the HbERF105 gene into a plant to improve the cold resistance of the plant.

[0012] The present invention also provides the use of the HbERF105 gene and the protein encoded by it, the recombinant vector or the recombinant bacteria in cultivating cold-resistant plant varieties.

[0013] The present invention also provides a method for cultivating cold-resistant plant varieties, comprising the steps of using genetic transformation technology to transfer the HbERF105 gene into a plant body to construct a transgenic cold-resistant plant with stably overexpressed HbERF105 gene.

[0014] The present invention also provides the use of a reagent for detecting the expression level of the HbERF105 gene in the early screening of cold-resistant plant varieties.

[0015] The present invention also provides application of the reagent for detecting the HbERF105 gene in cultivating cold-resistant plant varieties.

[0016] In the present invention, the above-mentioned plants include but are not limited to Arabidopsis thaliana and rubber tree.

[0017] The present invention discloses the following technical effects:

[0018] (1) This invention cloned for the first time a rubber tree ERF-type transcription factor gene HbERF105. The gene DNA sequence has no introns, an open reading frame of 825 bp, and encodes 274 amino acids, laying a theoretical foundation for the subsequent research and application of this gene.

[0019] (2) The present invention confirms that the HbERF105 gene of rubber trees is related to cold resistance and can actively respond to low temperature stress. It also discloses the expression pattern of the gene in rubber trees and its response mode under low temperature stress, providing technical support for the establishment of a method for screening cold-resistant varieties of rubber trees.

[0020] (3) The present invention confirms that in the study of rubber tree genetic breeding, the use of transgenic technology to overexpress the HbERF105 gene in plants can accelerate the plant's adaptation to the environment, increase the plant's survival rate, and effectively improve its cold resistance, which has a positive effect on improving the cold resistance of varieties, cultivating new varieties, and accelerating the breeding process. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0022] Figure 1 The figure is the result of subcellular localization of HbERF105 protein; the arrow points to the position of HbERF105 protein;

[0023] Figure 2 This is a statistical diagram of the expression pattern of HbERF105 in leaves of different varieties of rubber trees under low temperature stress;

[0024] Figure 3 The figure shows the PCR identification results of the transgenic Arabidopsis T2 generation strains; OE1-OE7 are the numbers of the transgenic Arabidopsis T2 generation strains, CK is the wild-type Arabidopsis strain, and M is a marker;

[0025] Figure 4 This is the phenotype of the growth status of Arabidopsis before low temperature stress;

[0026] Figure 5 This is the phenotype diagram of the growth status of Arabidopsis thaliana when low temperature stress is completed;

[0027] Figure 6 This is the phenotype of the growth status of Arabidopsis thaliana after 7 days of low temperature stress;

[0028] Figure 7 This is the phenotype of the growth status of Arabidopsis thaliana after 14 days of low temperature stress. DETAILED DESCRIPTION

[0029] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0030] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0031] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0032] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0033] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0034] The technical solution of the present invention is as follows:

[0035] The present invention clones a rubber tree ERF transcription factor gene HbERF105 from rubber tree variety 93114, analyzes the structural composition of the gene, and determines that the HbERF105 protein is localized in the nucleus by subcellular localization technology. The present invention also treats rubber tree seedlings with low temperature stress, detects the expression pattern of the HbERF105 gene in the seedlings under low temperature stress, and reveals that the HbERF105 gene may be related to the cold resistance of rubber trees.

[0036] In order to verify the regulatory effect of the HbERF105 gene on the cold resistance of rubber trees, the present invention uses the model organism Arabidopsis thaliana as the recipient plant, constructs a HbERF105 transgenic overexpression recombinant vector and a transgenic overexpression positive plant, and subjects the transgenic overexpression positive plant to low temperature stress treatment, confirming that the overexpression of the HbERF105 gene can improve the adaptability of plants to low temperature environments.

[0037] The method for constructing the recombinant vector of the present invention can be carried out by methods known in the art.

[0038] In some embodiments of the present invention, the method for constructing the recombinant vector is any of the following methods:

[0039] Enzyme ligation method, homologous recombination method, overlap extension PCR method.

[0040] In some embodiments of the present invention, the recombinant vector is constructed by enzyme ligation.

[0041] In the present invention, the expression pattern of the HbERF105 gene can be detected by methods known in the art.

[0042] In some embodiments of the present invention, the detection reagent used to detect the expression pattern of the HbERF105 gene is a reagent using any of the following methods:

[0043] polymerase chain reaction, nucleic acid hybridization, immunological detection and mass spectrometry.

[0044] The polymerase chain reaction can be further selected from fluorescent quantitative PCR (which can be combined with TaqMan probe for detection) and digital PCR;

[0045] The nucleic acid hybridization method can be further selected from in situ hybridization and transcriptome chip, etc.;

[0046] The immunological detection method can be further selected from WB, IP, IHC and antibody chip.

[0047] In some embodiments of the present invention, the detection reagent is a fluorescent quantitative PCR for detecting the gene expression.

[0048] In the embodiments of the present invention, the sources of the biological materials involved are as follows:

[0049] The vector pCAMBIA1300-GFP was purchased from Beijing Haimaboer Biotechnology Co., Ltd.; the vector pXCS-HAStrep was preserved by the National Rubber Tree Breeding Center and recorded in the literature "Zhai Qilin, An Zewei, Li Yachao, et al. Construction and application of a plant double gene co-expression vector [J]. Journal of Agricultural Biotechnology, 2013";

[0050] Agrobacterium EHA105 and GV3101 (MP90RK) were purchased from Beijing Coolbo Technology Co., Ltd.

[0051] Example 1 Cloning of HbERF105 gene

[0052] Based on the transcriptome data and genome information of rubber trees, the HbERF105 gene cloning primers ERF105-F and ERF105-R were designed. The cloning process is as follows:

[0053] The genomic DNA of rubber tree variety 93114 was extracted and amplified using ERF105-F and ERF105-R as primers.

[0054] ERF105-F: ATGGCACCACACGAAGCTTCAACCTTGG (SEQ ID NO. 1);

[0055] ERF105-R: CACAACCATGAGCCTAGAATA TCCCATCG (SEQ ID NO. 2).

[0056] Amplification system: 0.6 μL DNA template, 5 μL 2×PCR MasterMix, 0.2 μL each of primers ERF105-F (10 μM) and ERF105-R (10 μM), and 4 μL sterile ultrapure water.

[0057] Amplification program: 95°C for 3 min; 95°C for 30 s, 53°C for 30 s, 72°C for 2 min, for a total of 30 cycles; 72°C for 10 min.

[0058] The amplified product was sequenced and the obtained sequence was shown in SEQ ID NO.3. The sequencing results showed that the gene DNA sequence had no introns, the open reading frame was 825 bp, encoding 274 amino acids, and the amino acid sequence was shown in SEQ ID NO.4.

[0059] HbERF105 gene:

[0060] Atggcaccacacgaagcttcaaccttggagctcattcagcaatacctcctcactgactccaatcttgatactttttgcaattcccataacttttttaaaactgaaacccaagaatttgattgctttccgccacagcgaccagagttcaagcctcaactggttcataaatcttcgagtttgagccagcggaaacccaccatgagtaaaatatcgatacctcctccagcaaatttgaaccccatcccacctcagcaacctgtagcagtacctgcaactgagaataacaggtcggattgtagtgaagaaaagcattacagaggcgtccgccgtaggccgtggggaaaatacgcagcggagattcgagatcctaataagaaaggcgctcgtgtgtggctagggacattcaataccgctatagaagctgcaaaagcttatgatagtgctgcctttagattgcgtggaagcaaggccatactcaatttccctcttgaaattgggaatttggattccgatcattcccagcaagaatctgaattgacagataataacagtgactacgctaattgtaataataataataataataagaaaaggaagattgaggagacggagagcttggaaatcagcaataatattattaataataataagatgataaaaacagaaaatccttcaccggagagggatcaagcgacggatcccttaacgccgtcgagttggaaggggttttgggatggggaagtgaaggggatatttaatgtaccgccgttatcgccgttatcgccgcacccttcgatgggatattctaggctcatggttgtgtga (SEQ ID NO.3);

[0061] HbERF105 protein:

[0062] mapheastleliqqylltdsnldtfcnshnffktetqefdcfppqrpefkpqlvhkssslsqrkptmskisipppanlnpippqqpvavpatennrsdcseekhyrgvrrrpwgkyaaeirdpnkkgarvwlgtfntai eaakaydsaafrlrgskailnfpleignldsdhsqqeseltdnnsdyancnnnnnnkkrkieetesleisnniinnnkmiktenpsperdqatdpltpsswkgfwdgevkgifnvpplsplsphpsmgysrlmvv* (seq ID NO.4).

[0063] Example 2 Subcellular localization of HbERF105

[0064] The HbERF105 gene was subcloned into the plant expression vector pCAMBIA1300-GFP. The subcloning process was as follows:

[0065] Primers ERF105-LF and ERF105-LR with Spe I and Kpn I restriction sites were designed. The genomic DNA of rubber tree variety 93114 was used as a template to clone the rubber tree gene HbERF105. The obtained gene sequence and the vector pCAMBIA1300-GFP plasmid were double-digested with Spe I and Kpn I, respectively. The target gene fragment and vector fragment after restriction digestion were recovered, and the target gene fragment and vector fragment were connected with T4 DNA ligase to subclone the HbERF105 gene into the plant expression vector pCAMBIA1300-GFP. The recombinant plasmid that was successfully cloned was named: pCAMBIA1300-HbERF105-GFP.

[0066] ERF105-LF: GG ACTAGTAT ATGGCACCACACGAAGCTTCA (SEQ ID NO. 5); the underlined part is the restriction enzyme cleavage site.

[0067] ERF105-LR: CGG GGTACC CACAACCATGAGCCTAGAA (SEQ ID NO. 6); the underlined part is the restriction enzyme cleavage site.

[0068] Enzyme digestion system: 20 μL template DNA, 5 μL 10×FD Green Buffer, 2.5 μL each of Spe I and Kpn I endonucleases, 20 μL sterile ultrapure water. Terminate the reaction after 5 min in a 37°C water bath, perform electrophoresis detection, and recover the enzyme-digested fragments.

[0069] Ligation system: 10×Buffer 1μL, pCAMBIA1300-GFP fragment 3μL, HbERF105 fragment 5μL, T4DNA ligase 1μL. Mix well and incubate in a 16℃ water bath for 16h, terminate the reaction, and transform the ligation product.

[0070] The recombinant plasmid pCAMBIA1300-HbERF105-GFP carrying the HbERF105 gene was transferred into Agrobacterium EHA105 and cultured until OD 600 When the OD value was about 0.8, the bacteria were collected and washed twice with a resuspension solution (1 / 2MS, 100 μL acetosyringone, 0.0012% L-77, pH = 5.7), and the OD value of the bacterial solution was adjusted. 600 The concentration of the bacterial solution was 0.8, and a 5mL sterile syringe was used to absorb the bacterial solution, and it was slowly injected into the tobacco leaves from the lower surface of the leaves until the tobacco leaves were moistened. The injected tobacco was first cultured in the dark for 24 hours, and then cultured normally for 2 days. The tobacco leaves at the injection position were soaked in DAPI (100ng / mL) nuclear dye for 1 hour, and the green fluorescence signal was observed under a fluorescence microscope. The results are as follows: Figure 1 As shown, the arrow points to the HbERF105 protein, which is located in the cell nucleus.

[0071] Example 3 Analysis of HbERF105 response to low temperature stress

[0072] One-year-old bagged budded seedlings of rubber tree varieties 93114 and Reyan 73397 with two stable leaves were subjected to low temperature stress. The bagged budded seedlings were transferred to an artificial climate chamber (16 h light / 8 h dark, 160 μmol·m -2 ·s -1The cells were cultured at 25 °C for two days in an oven (100 °C for 24 h, 80% light intensity, and 80% relative humidity), and then the temperature was lowered to 4 °C to simulate low temperature stress. Samples were taken at 0 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, and 48 h after treatment, and total RNA was extracted using RNAprep pure Polysaccharide and Polyphenol Plant Total RNA Extraction Kit (TIANGEN) according to the instructions. After reverse transcription into cDNA, the HbActin gene of rubber tree was used as the internal reference gene, and internal reference primer pairs ACT-QF and ACT-QR were designed. qRT-PCR was performed with gene primer pairs ERF105-QF and ERF105-QR to analyze the expression of HbERF105 in response to low temperature stress.

[0073] ERF105-QF: AAGATTGAGGAGACGGACAGC (SEQ ID NO.7);

[0074] ERF105-QR: ATCCCAAAACCCTTCCAACT (SEQ ID NO.8);

[0075] ACT-QF: CAGTGGTCGTACAACTGGTAT (SEQ ID NO.9);

[0076] ACT-QR: ATCCTCCAATCCAGACACTGT (SEQ ID NO. 10).

[0077] The cDNA synthesis system is as follows: add 3μg RNA, 1μL random primer, and 8μL sterile ultrapure water to a 200μL PCR tube, mix well, denature in a 65℃ water bath for 5min, and then place in an ice bath; then add 4μL 5× Reaction buffer, 1μL 20U / μLRNase Inhibitor, 2μL 10mM dNTPs, and 1μL 200U / μL reverse transcriptase in sequence, mix well, and incubate at 25℃ on a PCR instrument for 5min, then incubate at 42℃ for 60min, and inactivate at 70℃ for 5min.

[0078] The reaction system of qRT-PCR was: 2 μL of cDNA, 10 μL of 2×Premix Ex Taq, 0.4 μL each of ERF105-QF (10 μM) and ERF105-QR (10 μM), and 7.2 μL of EASY Dilution.

[0079] The reaction program was: 95°C for 30 s; 95°C for 5 s, 60°C for 20 s, and 72°C for 20 s, for a total of 45 cycles.

[0080] The results are as follows Figure 2As shown, the HbERF105 gene was induced to express under low temperature stress, but the degree of induced expression in the cold-tolerant variety 93114 was stronger than that in the cold-intolerant variety Reyan 73397. In addition, its response speed to low temperature stress was also faster, and the expression level increased significantly after 0.5 h of stress.

[0081] Example 4 HbERF105 Functional Verification

[0082] The HbERF105 gene was subcloned into the plant expression vector pXCS-HAStrep, and the cloning process was as in Example 2. The successfully cloned recombinant plasmid was named: pXCS-HbERF105-HAStrep. The pXCS-HbERF105-HAStrep plasmid was then transformed into Agrobacterium GV3101 (MP90RK) and cultured until OD 600 When the concentration was about 0.8, the bacteria were collected and the infection solution (4.4 g / L MS, 10 μL / mL 6-BA, 5% sucrase, 0.03% MES and 0.0012% L-77) was added to suspend the bacteria. The prepared infection solution containing Agrobacterium was placed in a sprayer and sprayed on the inflorescence of the quasi-ecotype Arabidopsis plants (Col-0 type). When liquid droplets continued to drip from the plants, the spraying was stopped. The infected plants were cultured in the dark for 1 day and then restored to normal light culture. The infection was carried out once every 2 days for a total of 3 treatments. After the treatment was completed, the seeds were watered normally and cultured until the seeds matured to obtain T1 generation seeds.

[0083] Sow T1 generation seeds on vermiculite. When Arabidopsis grows two true leaves, spray 10 mg / L glufosinate solution to screen transgenic Arabidopsis positive plants. Treat once every 3 days. Transgenic Arabidopsis plants that survive 3 treatments are positive plants. The seeds of positive plants collected from a single plant are the T2 generation. Select T2 generation transgenic Arabidopsis strains with a survival and death ratio that meets the 3:1 separation ratio, numbered OE1-OE7, transplanted to vermiculite, and after the plants grow for 2 weeks, perform PCR identification. The PCR identification process is as follows:

[0084] The genomic DNA of the leaves of wild-type and transgenic Arabidopsis plants was extracted, and the transgenic genomic DNA was used as the test template, and the genomic DNA of the leaves of wild-type Arabidopsis plants was used as the control template (CK). The primer pair ERF105-F (SEQ ID NO.1) and ERF105-R (SEQ ID NO.2) were used for PCR amplification. The amplification system and amplification procedure were the same as those in Example 1. The amplified product was detected by gel electrophoresis. The results were as follows: Figure 3 As shown, when a specific band of 825 bp in size is detected in the template to be tested, the plant is determined to be a T2 generation positive transgenic Arabidopsis strain.

[0085] T3 seeds were harvested from qualified individual plants. T3 pure lines that no longer separated were selected for HbERF105 gene function analysis, and three pure lines L1, L2, and L3 were obtained.

[0086] After the T3 generation Arabidopsis plants grew for 28 days, all materials and wild-type (CK) Arabidopsis plants were treated at -5°C for 2 days for low temperature stress, and then restored to normal environment cultivation. The plant survival rate was counted on the 7th and 14th days, and the phenotype was observed. The plant survival rate results are shown in Table 1, and the phenotypic status is shown in Table 1. Figure 4-Figure 7 As shown, it can be seen that the transgenic strains not only have a high survival rate, but also recover normal growth quickly, showing strong cold resistance.

[0087] Table 1 Survival rate of different strains of Arabidopsis thaliana after low temperature stress

[0088]

[0089] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of the rubber tree ERF transcription factor HbERF105 gene and the protein encoded therein, a recombinant vector containing the rubber tree ERF transcription factor HbERF105 gene, or a recombinant bacterium containing the rubber tree ERF transcription factor HbERF105 gene in regulating plant cold resistance, characterized in that: The regulating plant cold tolerance is to improve the plant cold tolerance by overexpressing the HbERF105 gene; The nucleotide sequence of the HbERF105 gene is shown in SEQ ID NO.3; the amino acid sequence of the protein is shown in SEQ ID NO.4; The plant is rubber tree or Arabidopsis thaliana.

2. A method for regulating plant cold tolerance, characterized in that: The method comprises the steps of using genetic transformation technology to transfer the rubber tree ERF transcription factor HbERF105 gene into the plant body to improve the cold resistance of the plant; The nucleotide sequence of the HbERF105 gene is shown in SEQ ID NO.3; the plant is rubber tree or Arabidopsis thaliana.

3. Application of the rubber tree ERF transcription factor HbERF105 gene and the protein encoded therein, a recombinant vector containing the rubber tree ERF transcription factor HbERF105 gene, or a recombinant bacterium containing the rubber tree ERF transcription factor HbERF105 gene in cultivating cold-resistant plant varieties, characterized in that: The cultivation of cold-resistant plant varieties is to construct transgenic cold-resistant plants with stable overexpression of the HbERF105 gene; The nucleotide sequence of the HbERF105 gene is shown in SEQ ID NO.3; the amino acid sequence of the protein is shown in SEQ ID NO.4; The plant is rubber tree or Arabidopsis thaliana.

4. A method for cultivating cold-resistant plant varieties, characterized in that: The method comprises the steps of using genetic transformation technology to transfer the rubber tree ERF transcription factor HbERF105 gene into the plant body to construct a transgenic cold-resistant plant with stably overexpressed HbERF105 gene; The nucleotide sequence of the HbERF105 gene is shown in SEQ ID NO.3; the plant is rubber tree or Arabidopsis thaliana.

5. Application of a reagent for detecting the expression of the ERF transcription factor HbERF105 gene in early screening of cold-resistant plant varieties, characterized in that: The nucleotide sequence of the HbERF105 gene is shown in SEQ ID NO.3; the plant is rubber tree or Arabidopsis thaliana; The HbERF105 gene is overexpressed in the plant.

6. Application of a reagent for detecting the ERF transcription factor HbERF105 gene of rubber tree in cultivating cold-resistant plant varieties, characterized in that: The nucleotide sequence of the HbERF105 gene is shown in SEQ ID NO.3; the plant is rubber tree or Arabidopsis thaliana; The HbERF105 gene is overexpressed in the plant.

Images