TaNAC67-like gene and its encoded protein and application

Abstract

The application belongs to the technical field of plant genetic engineering, and relates to a TaNAC67-Like gene, a coding protein thereof and application. The application provides the TaNAC67-Like gene, wherein the TaNAC67-Like gene: a) codes the TaNAC67-Like protein; and b) has a nucleotide sequence as shown in SEQ ID NO:1. The application explicitly defines the positive regulation function of the TaNAC67-Like gene in drought resistance, and provides a brand-new gene resource and technical idea for wheat drought resistance trait improvement, drought resistance molecular breeding and high-quality drought-resistant wheat variety cultivation.

Description

Technical Field

[0001] This invention belongs to the field of plant genetic engineering technology, and relates to the TaNAC67-Like gene and its encoded protein and its applications. Background Technology

[0002] Wheat (Triticum aestivum) is one of the three major food crops, and its stable and high yield is directly related to food security and the stable development of the agricultural economy. Against the backdrop of global climate change, the frequency and impact of extreme weather disasters such as drought are increasing year by year, and their scope is continuously expanding. These have become core abiotic stress factors restricting wheat growth and development, and reducing wheat yield and quality. Statistics show that approximately 26.7% of my country's existing wheat-growing area suffers from drought stress year-round. Severe drought can cause significant yield reductions or even crop failure, greatly hindering large-scale, high-quality, and stable wheat production. Therefore, identifying efficient and stable drought-resistant functional genes in wheat, elucidating their molecular regulatory mechanisms of drought resistance, and breeding new drought-resistant wheat varieties are urgent technical challenges to be solved in the field of wheat stress-resistance breeding.

[0003] NAC transcription factors (NAM, ATAF1 / 2, CUC2) are an important family of transcription factors unique to plants. Members of this family are widely involved in a series of key physiological processes, including plant growth and development regulation, secondary metabolism synthesis, and responses to biotic and abiotic stresses. They are core regulatory factors in the molecular regulatory network of plant stress resistance. Existing molecular mechanism studies have confirmed that NAC transcription factors participate in plant stress resistance responses through a hierarchical regulatory model. They can directly activate MYB46 and MYB83, and synergistically activate the expression of downstream functional genes such as MYB58, MYB63, and MYB85, as well as key genes for lignin biosynthesis, thereby regulating plant cell wall synthesis and remodeling processes and enhancing plant stress tolerance.

[0004] Existing crop stress resistance studies have confirmed the conserved function of NAC transcription factors in regulating lignin synthesis and mediating abiotic stress responses. In rice, OsNAC17 positively regulates the expression of lignin synthesis-related genes, promoting the accumulation of lignin in leaves and root tissues and significantly enhancing the drought tolerance of rice plants. In celery, AgNAC1 can regulate the transcriptional levels of key genes in the lignin synthesis pathway, altering the efficiency of lignin biosynthesis and the structure of cell wall components, thereby improving celery's adaptability to salt stress. These studies fully demonstrate that the participation of NAC family genes in plant abiotic stress responses through the regulation of lignin synthesis pathways is a conserved regulatory mechanism in the plant kingdom, possessing broad potential for breeding applications.

[0005] Although existing research has confirmed the important role of NAC transcription factors in crop stress resistance regulation, the exploration of functional genes of the NAC family in wheat that can specifically regulate drought stress response remains insufficient. In particular, the drought resistance regulatory function, molecular mechanism, and breeding application of the TaNAC67-Like homolog have not yet been developed or reported, and there is still a gap in high-quality gene resources for wheat drought resistance molecular breeding. Therefore, exploring and identifying the drought resistance function of wheat TaNAC67-Like genes and elucidating their molecular mechanism of regulating wheat drought stress response can provide new theoretical basis for the study of wheat drought resistance molecular mechanisms. At the same time, it can provide new gene resources and technical support for creating new drought-resistant wheat germplasm and breeding new drought-resistant and high-yielding wheat varieties, which has important theoretical research value and agricultural production application value. Summary of the Invention

[0006] To address the shortcomings of existing wheat drought resistance improvement technologies, this invention provides the TaNAC67-Like gene and its novel application in regulating wheat drought resistance.

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] On one hand, the present invention provides a TaNAC67-Like gene, wherein the TaNAC67-Like gene: a) Encodes the TaNAC67-Like protein; b) The open reading frame has a nucleotide sequence as shown in SEQ ID NO:1.

[0009] On the other hand, the present invention provides a TaNAC67-Like protein having an amino acid sequence as shown in SEQ ID NO:2.

[0010] It also provides recombinant expression vectors, expression cassettes, transgenic cell lines or recombinant strains containing the TaNAC67-Like gene as described in this invention.

[0011] The present invention also provides the application of the TaNAC67-Like gene in the creation of TaNAC67-Like gene RNAi materials.

[0012] The invention also provides the application of the TaNAC67-Like gene in regulating wheat drought resistance, including: overexpressing the TaNAC67-Like gene to improve wheat drought resistance.

[0013] The present invention also provides the application of the TaNAC67-Like protein in regulating wheat drought resistance.

[0014] The invention also provides the application of the recombinant expression vector, expression cassette, transgenic cell line or recombinant strain described in this invention in regulating wheat drought resistance.

[0015] A method for breeding drought-resistant wheat varieties is also provided, comprising: overexpressing the TaNAC67-Like gene described in this invention in wheat plants to obtain TaNAC67-Like gene overexpressing wheat plants.

[0016] Furthermore, the TaNAC67-Like gene plays a positive regulatory role in wheat drought resistance.

[0017] A method for cultivating drought-resistant transgenic plants is also provided, the method comprising the following steps: S1. Increase the expression of the TaNAC67-Like gene in the target plant to obtain transgenic plants; S2. Increase the content and / or activity of TaNAC67-Like protein in the target plant to obtain transgenic plants; among which, The transgenic plant exhibits higher drought resistance than the target plant; The plant in question is wheat.

[0018] A method for cultivating drought-resistant transgenic plants is also provided, the method comprising the following steps: To increase the expression of the TaNAC67-Like gene in target plants to obtain transgenic plants; and / or By increasing the content and / or activity of TaNAC67-Like protein in target plants, transgenic plants are obtained; among them, The transgenic plant exhibits higher drought resistance than the target plant; The plant in question is wheat.

[0019] SEQ ID NO:1 is as follows:

[0020] SEQ ID NO: 2 is specifically as follows: MIDPQANLDELYCIWLPNTTLSVTGESTGTTTPCASLAVRAVYKSSDYYPGTESSLSQEESDATPHLDLKPRLAVDRGGPGAGGRPAGRTCKSWMGNYVFPLNLPPGYHFAPSGDELIVHYLRRKLDGLPPHLPIFNDVTPIIDYRPEQITDEFRDFGEGARWYFFTKRTRKYATGSRPD RTTPGNGFWKATGPVREIPICPGGKLVGRARTLVFYTGPEKPTHWTMYEYENRTSEAEAN DKNVDKLGEWVLCTIQKSQRCDQKKEAGGKAKSKAGGKKRKGKARNGADLVQIQSAEVPEIAWPEKVDEAYVFDDLEQTGCPKKRPQIEQHEPPRSETMMVDDYDYNGIQYIWDDEYMPPPPLPLPLEYEVAPHVAGVGYSYNNMMYQPANSYLHAAHYAVEQDGYAGGHLGSLLGTGNIPCAYRPQHSCAAGITNLPWSRGATTSGQVEP.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention clarifies that the TaNAC67-Like gene, with the nucleotide sequence shown in SEQ ID NO:1, is a positively regulating gene in the wheat drought stress response process. It elucidates for the first time the molecular regulatory mechanism of the TaNAC67-Like protein in drought resistance, confirming that this protein can specifically bind to the promoter of the lignin synthesis gene TaCYP450 and activate its expression. This improves the molecular regulatory network of wheat drought resistance and fills the technical gap in the prior art where the drought resistance regulatory mechanism of this gene was unclear. Furthermore, relying on VIGS transient silencing technology, this invention clearly demonstrates, from the perspective of physiological indicators such as plant relative water content and proline content, that silencing the TaNAC67-Like gene significantly weakens the drought resistance of wheat plants. Furthermore, by constructing stable overexpression vectors and RNAi interference vectors, this invention successfully created drought-resistant lines with TaNAC67-Like gene overexpression and gene silencing, obtaining stable wheat drought-resistant germplasm materials. This provides reliable experimental materials, novel functional genes, and comprehensive theoretical support for subsequent in-depth research on the molecular mechanisms of wheat drought resistance, targeted improvement of drought resistance traits, and breeding of new drought-resistant varieties. It has significant scientific research value and application prospects for enriching wheat stress-resistance breeding gene resources and improving wheat's adaptability to drought stress. Attached Figure Description

[0022] Figure 1 Phylogenetic tree diagram of the TaNAC67-Like gene.

[0023] Figure 2 This is an image showing the electrophoresis results of the TaNAC67-Like gene. M represents the DNA molecular weight marker; lanes 1 and 2 represent the amplified products of the TaNAC67-Like specific fragment.

[0024] Figure 3 This is a diagram analyzing the gene expression patterns of the TaNAC67-Like gene under drought stress. Different lowercase letters represent the expression patterns observed under Duncan's multiple comparison test. P At the level <0.05, the differences between groups are significant; the same lowercase letter indicates no significant difference between groups.

[0025] Figure 4 This is a comparative diagram of drought resistance phenotypes in wheat plants after transient silencing of the TaNAC67-Like gene using VIGS. A represents phenotypic verification of successful VIGS silencing system construction; B represents phenotypic identification of silent plants after drought stress treatment; C represents the determination of silencing efficiency, proline content, and survival rate of silent plants. represent P The difference was statistically significant at levels <0.05. represent P The difference at the <0.01 level was highly significant.

[0026] Figure 5 This figure shows the results of drought resistance phenotype identification between T2 generation TaNAC67-Like overexpressing lines and RNAi stably silenced lines. WT represents the wheat variety Fielder; TaNAC67L-OE represents overexpressing wheat plants; and TaNAC67L-RNAi represents silenced wheat plants.

[0027] Figure 6 This is a graph validating the results of the EMSA experiment. Lane 1 represents the negative control of the MBP-tagged protein (MBP) and the labeled probe (promoter-Probe); Lane 2 shows that the TaNAC67L-MBP fusion protein can bind to the labeled probe to form a migration band; Lanes 3 and 4 represent the binding reaction of the TaNAC67L-MBP fusion protein to the labeled probe after the addition of an unlabeled competing probe; Shift band represents the migration band; Free Probe represents the free probe without protein binding. Detailed Implementation

[0028] The technical solution of the present invention will be described below with reference to embodiments. However, the present invention is not limited to the following embodiments. Unless otherwise specified, the experimental methods and detection methods described in each embodiment are conventional methods; unless otherwise specified, the reagents and materials can be purchased commercially. Furthermore, unless otherwise specified, the percentage content and material ratios involved in the embodiments refer to mass percentage content and mass ratio.

[0029] Figure 1 Phylogenetic tree diagram of the TaNAC67-Like gene.

[0030] Figure 2 This is an image showing the electrophoresis results of the TaNAC67-Like gene. M represents the DNA molecular weight marker; lanes 1 and 2 represent the amplified products of the TaNAC67-Like specific fragment, with a corresponding fragment size of 1356 bp, and two biological replicates are set up; the specific amplified fragment of the TaNAC67-Like gene is 1356 bp in size.

[0031] Figure 3 This is a diagram analyzing the gene expression patterns of the TaNAC67-Like gene under drought stress. Different lowercase letters represent the expression patterns observed under Duncan's multiple comparison test. P At the level <0.05, the differences between groups are significant; the same lowercase letter indicates no significant difference between groups.

[0032] Figure 4This is a comparison of drought resistance phenotypes in wheat plants after transient silencing of the TaNAC67-Like gene using VIGS, confirming that silencing the TaNAC67-Like gene can significantly reduce the drought resistance of wheat plants. A represents the phenotypic verification of the successful construction of the VIGS silencing system; wheat leaves infected with BSMV-PDS showed photobleaching symptoms, while wheat leaves infected with BSMV-γ and BSMV-TaNAC67L showed streaky chlorosis, while the control leaves showed no changes; B represents the phenotypic identification of silent plants after drought stress treatment; C represents the determination of the silencing efficiency, proline content, and survival rate of silent plants; MOCK represents the blank infection control group; BSMV-PDS represents the positive control of the VIGS system; BSMV-γ represents the BSMV empty negative control group; BSMV-TaNAC67L represents the TaNAC67-Like gene silencing treatment group; BSMV:TaNAC67L#1, BSMV:TaNAC67L#2, and BSMV:TaNAC67L#3 represent three independent biological duplicates of the TaNAC67-Like gene silencing treatment, respectively; represent P The difference was statistically significant at levels <0.05. represent P The difference at the level <0.01 is extremely significant; BSMV:γ in C refers to the same term as BSMV-γ in A.

[0033] Figure 5 This figure shows the results of drought resistance phenotype identification between T2 generation TaNAC67-Like overexpressing lines and RNAi stably silenced lines. WT represents the wheat variety Fielder; TaNAC67L-OE represents overexpressing wheat plants; and TaNAC67L-RNAi represents silenced wheat plants.

[0034] Figure 6 This image shows the EMSA experimental results, demonstrating that the TaNAC67-Like protein specifically binds to the promoter of the lignin synthesis gene TaCYP450. Lane 1 represents the negative control (MBP-tagged protein and labeled probe, promoter-probe) with no specific binding band. Lane 2 shows the TaNAC67L-MBP fusion protein binding to the labeled probe, forming a migration band. Lanes 3 and 4 represent the binding reaction of the TaNAC67L-MBP fusion protein to the labeled probe after the addition of an unlabeled competing probe. In lanes 3-4, the migration band signal gradually weakens with the addition of 20× and 100× unlabeled competing probes, indicating that the binding of TaNAC67L to the promoter is sequence-specific. Shift band represents the migration band; Free Probe represents the free probe without bound protein.

[0035] Example 1 This embodiment provides sequence analysis of the TaNAC67-Like gene.

[0036] In this embodiment, the wheat TaNAC67-Like gene was obtained using open reading frame (ORF) cloning technology. Sequencing confirmed that the nucleotide sequence of the open reading frame of this gene is shown in SEQ ID NO:1, with a full length of 1356 bp. The TaNAC67-Like protein encoded by the TaNAC67-Like gene has the amino acid sequence shown in SEQ ID NO:2. Physicochemical analysis of the protein showed that the molecular weight of the protein encoded by this gene is approximately 50.1 kDa, and the isoelectric point is 5.67. Predictive analysis of the conserved domains of the TaNAC67-Like protein using the Uniprot database showed that the TaNAC67-Like protein contains typical NAM conserved domains, thus identifying it as a member of the NAC transcription factor family. The homologous gene sequences of TaNAC67-Like were compared using MEGA 7.0 software, and a phylogenetic tree was constructed. The evolutionary analysis results showed that wheat TaNAC67-Like (see...) Figure 1 The transcription factor *Triticum dicoccoides* NAC 67-L is most closely related to the *Erythromyces cerevisiae* TdNAC67-L transcription factor, and has the smallest evolutionary distance. Figure 1 Based on the above gene structure, conserved domains, and evolutionary characteristics, this invention names this wheat gene the TaNAC67-Like gene.

[0037] Example 2 This embodiment provides the expression pattern of the TaNAC67-Like gene.

[0038] To clarify whether the TaNAC67-Like gene is involved in the drought stress response process in wheat, this embodiment uses Xindong 18 wheat as the experimental material for stress expression analysis. Wheat seeds were planted in a greenhouse soil environment with a culture temperature of 22℃-25℃, a light duration of 16 h / day, and a relative humidity of 60%-70%, and were conventionally cultured until the wheat reached the two-leaf-one-heart growth stage. Fresh wheat leaf samples were randomly collected, flash-frozen in liquid nitrogen, and then cryopreserved at -80℃. Total RNA was then extracted from the leaves and reverse transcribed to obtain a cDNA template for cloning the TaNAC67-Like gene. Figure 2 ).

[0039] Simultaneously, wheat seedlings with uniform growth, robust development, and free from pests and diseases (two leaves and one heart) were selected and divided into a control group and a drought stress treatment group. The treatment group was subjected to drought stress treatment with a 20% PEG (polyethylene glycol) solution. Using the TaEF-1α gene as an internal reference gene, the expression differences of the TaNAC67-Like gene under drought stress conditions were detected and analyzed using real-time quantitative PCR. Experimental data were analyzed using 2... -ΔΔCT Statistical analysis was conducted to clarify the response characteristics of the TaNAC67-Like gene to wheat drought stress.

[0040] Depend on Figure 3 The expression characteristics of the TaNAC67-Like gene in the wheat variety "Xindong 18" under drought stress were detected using real-time quantitative PCR (qRT-PCR). The mRNA expression level of this gene reached its highest value after 1 h of drought stress treatment, representing an upregulation of approximately 13.6-fold compared to 0 h. This confirms that the TaNAC67-Like gene participates in the drought stress response of wheat "Xindong 18" and plays a functional role in the plant's drought stress response.

[0041] Example 3 This embodiment explores the functional role of silencing the TaNAC67-Like gene in the wheat drought stress response process.

[0042] Construction of TaNAC67-Like gene-silenced wheat materials The specific sequence of the TaNAC67-Like gene, TaNAC67-Like-as, was obtained and amplified to successfully construct the recombinant gene silencing vector BSMV:TaNAC67-Like-as. The recombinant gene silencing vector, the positive control vector BSMV-PDS, and the empty plasmids BSMV:α, BSMV:β, and BSMV:γ (not shown in the figure) were linearized. In vitro transcription was performed on all linearized vectors to obtain the corresponding RNA transcripts. These transcripts were then inoculated into wheat seedlings using a virus-inoculation method, completing the creation of wheat materials with transient TaNAC67-Like gene silencing. After successful virus inoculation in the wheat plants, drought stress resistance identification experiments were conducted. Ten days after inoculation with the virus, functional leaf samples of wheat were collected, and the silencing efficiency of the TaNAC67-Like gene was detected by real-time quantitative PCR (qRT-PCR) to verify the gene silencing effect. Fourteen days after inoculation with the virus, wheat seedlings with consistent growth, height, and growth status were selected, and leaf tissue samples were collected to determine drought-related physiological indicators such as relative water content and proline content.

[0043] The TaNAC67-Like-as sequence is as follows: CAGAGACAATGATGGTAGACGACTATGACTACAATGGTATCCAATACATCTGGGATGATGAGTACATGCCGCCGCCGCCGCTGCCGCTGCCGCTGGAGTATGAGGTGGCGCCTCATGTTGCAGGAGTAGGATACAGCTACAACAACATGATGTATCAGCCGGCTAACAGCTACCTACATG.

[0044] Depend on Figure 4 The gene silencing efficiency test results showed that, using wheat plants inoculated with the BSMV-γ empty vector as a blank control, the mRNA expression level of the TaNAC67-Like gene was significantly downregulated in wheat plants inoculated with the BSMV:TaNAC67-Like recombinant vector (BSMV-TaNAC67), with a silencing efficiency of 39%-44%. This confirms that the VIGS silencing system in this embodiment is effective and can achieve specific silencing of the TaNAC67-Like gene. The wheat drought stress resistance test results showed that, after transient silencing of the TaNAC67-Like gene mediated by BSMV, the survival rate of wheat seedlings under drought stress significantly decreased to 35%-40%. Simultaneously, physiological index tests showed that the proline content in the leaves of TaNAC67-Like gene-silenced plants was significantly lower than that of the blank control plants.

[0045] Example 4 This embodiment further verifies the biological function of the TaNAC67-Like gene in the wheat drought stress response process by constructing transgenic wheat materials with TaNAC67-Like gene overexpression and RNAi interference.

[0046] Based on the CDS sequence of the wheat TaNAC67-Like gene, the pCub-TaNAC67-Like overexpression vector was constructed using gene cloning technology. Simultaneously, specific amplification primers were designed and synthesized from the conserved 200 bp-400 bp region of the TaNAC67-Like gene. Using Gateway cloning technology, the gene-specific interference fragment was precisely inserted into the pBWA(V)BU vector, completing the construction of the TaNAC67-Like gene RNAi interference vector. The successfully constructed recombinant vector was introduced into *Agrobacterium tumefaciens* EHA105 competent strain. Through an *Agrobacterium*-mediated genetic transformation system, immature embryos of Fielder wheat were transformed to obtain transgenic wheat positive plants.

[0047] For the transgenic wheat lines obtained through transformation, positive screening was performed using a combination of Bar test strip detection and PCR molecular identification to obtain stably inherited transgenic lines. Subsequent RT-qPCR data for quantitative gene expression detection are expressed as the mean ± standard error (SE) of three independent biological samples. T2 generation TaNAC67-Like overexpression homozygous transgenic plants were selected as the experimental group, with wild-type Fielder (WT) wheat plants serving as the blank control group. Both groups were simultaneously cultured under uniform and suitable environmental conditions. When the plants reached the two-leaf-one-heart seedling stage, both groups were uniformly subjected to 20% PEG drought stress treatment. To ensure experimental reproducibility and data reliability, three biological replicates were set up for each group, with 10 uniformly growing and healthy wheat plants planted in each replicate. Throughout the process, the light, temperature, humidity, and other growth environmental conditions, as well as the drought stress treatment conditions, were strictly controlled to ensure the uniqueness of experimental variables.

[0048] Depend on Figure 5 Based on the results of Bar test strip detection and qRT-PCR quantitative verification, plants propagated to the T2 generation were used as the core experimental material for drought resistance verification experiments. After 7 days of continuous drought stress treatment, phenotypic observations showed that under normal and suitable growth conditions, there were no significant differences in plant type, leaf color, growth rate, and other morphological characteristics between the TaNAC67-Like gene overexpressing transgenic plants and wild-type Fielder plants. Under continuous drought stress treatment, wild-type wheat plants showed more pronounced leaf wilting and yellowing, while the TaNAC67-Like-OE transgenic plants (TaNAC67L-OE) exhibited significantly milder drought symptoms, with leaves maintaining a better green state, and significantly better water retention and drought resistance than wild-type plants. Conversely, the RNAi interference line (TaNAC67L-RNAi) showed obvious leaf curling and wilting after drought stress treatment, with some leaves turning yellow, and the degree of wilting was significantly more severe than that of wild-type (WT) plants. This indicates that the TaNAC67-Like gene plays a positive regulatory role in wheat drought stress.

[0049] Example 5 This embodiment uses gel migration arrest assay (EMSA) to verify the in vitro specific binding relationship between TaNAC67-Like protein and TaCYP450 gene promoter.

[0050] The MBP vector backbone was linearized using a double digestion system of NdeI and BamHI. To achieve precise homologous recombination ligation between the target fragment and the MBP vector backbone, an NdeI restriction linker sequence was introduced at the end of the upstream primer for MBP vector backbone amplification. The specific linker sequence was: 5'-GGAAGGATTTCACATATGATGATTGATCCACAAGCAA-3'; and a BamHI restriction linker sequence was introduced at the end of the downstream primer. The specific linker sequence was: 5'-TGCAGGGAATTCGGATCCCGGCTCTACCTGCCCGCTC-3'. Simultaneously, a probe named GATCATGGGATGCGTGGATGGGCCATG was designed for the cloning and amplification of the wheat TaCYP450 gene promoter fragment.

[0051] The TaCYP450 gene sequence is as follows:

[0052] like Figure 6 As shown, based on the conserved ABRE motif in the TaCYP450 gene promoter sequence, biotin-labeled specific probes were designed using 10 bp sequence fragments upstream and downstream of this motif. Simultaneously, an unlabeled homologous sequence identical to the labeled probe was used as a cold competitive probe for specific competitive control. Prokaryotic expression and purification of the MBP-TaNAC67-Like fusion protein were also performed to obtain high-purity experimental protein samples for subsequent EMSA interaction verification experiments. This embodiment strictly followed the standard operating procedures of the EMSA kit for probe-protein binding reactions and electrophoresis detection. Experimental results showed that in the blank lane (lane 1) with only the biotin-labeled probe and no TaNAC67-Like protein, only a fast-migrating free probe band was detected, with no specific binding band generated. After adding purified TaNAC67-Like protein to the system, the labeled probe containing the ABRE core element of CGTG could specifically bind to the TaNAC67-Like protein, resulting in a significantly delayed DNA-protein complex specific binding band in the electrophoresis pattern (lane 2). Furthermore, gradient dilutions of the unlabeled cold competitive probe and competitive binding experiments showed that as the concentration of the cold competitive probe gradually decreased, the binding band intensity of the DNA-protein complex continuously weakened, and the competitive binding effect exhibited a concentration-dependent change (lanes 3 and 4). In summary, the EMSA results confirm that the TaNAC67-Like protein can directly and specifically bind to the CGTG core ABRE element in the promoter region of the TaCYP450 gene.

[0053] As described above, the basic principles, main features, and advantages of the present invention have been well described. The above embodiments and specifications are merely descriptions of preferred embodiments of the present invention, and the present invention is not limited to the above embodiments. Various changes and improvements made to the technical solutions of the present invention by those skilled in the art without departing from the spirit and scope of the present invention should fall within the protection scope defined by the present invention.

Claims

1. The TaNAC67-Like gene, characterized by, The TaNAC67-Like gene: a) Encodes the TaNAC67-Like protein; b) The open reading frame has a nucleotide sequence as shown in SEQ ID NO:

1.

2. TaNAC67-Like protein, characterized in that, The TaNAC67-Like protein has the amino acid sequence shown in SEQ ID NO:

2.

3. A recombinant expression vector, expression cassette, transgenic cell line, or recombinant strain containing the TaNAC67-Like gene as described in claim 1.

4. The application of the TaNAC67-Like gene as described in claim 1 in the creation of TaNAC67-Like gene RNAi materials.

5. The application of the TaNAC67-Like gene as described in claim 1 in regulating wheat drought resistance, characterized in that, include: Overexpression of the TaNAC67-Like gene improves drought resistance in wheat.

6. The application of the TaNAC67-Like protein according to claim 2 in regulating wheat drought resistance.

7. The application of the recombinant expression vector, expression cassette, transgenic cell line or recombinant strain as described in claim 3 in regulating wheat drought resistance.

8. A method for breeding drought-resistant wheat varieties, characterized in that, include: Overexpressing the TaNAC67-Like gene as described in claim 1 in wheat plants yields TaNAC67-Like gene overexpressing wheat plants.

9. The method according to claim 8, characterized in that, The TaNAC67-Like gene plays a positive regulatory role in wheat drought resistance.

10. A method for cultivating drought-resistant transgenic plants, characterized in that, The method includes the following steps: S1. Increase the expression of the TaNAC67-Like gene in the target plant to obtain transgenic plants; S2. Increase the content and / or activity of TaNAC67-Like protein in the target plant to obtain transgenic plants; among which, The transgenic plant exhibits higher drought resistance than the target plant; The plant in question is wheat.

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