Camellia sinensis csnac72 gene, protein and application thereof in breeding of resistant variety and regulating of anthracnose susceptibility

Abstract

This invention discloses the CsNAC72 gene and protein of tea plants, and their application in breeding anthracnose-resistant varieties and regulating anthracnose susceptibility, belonging to the field of genetic engineering technology. The nucleotide sequence of the CDS region of this gene is shown in SEQ ID NO.1. This invention further provides the application of this gene in regulating anthracnose resistance / susceptibility in tea plants and constructs anthracnose-resistant and susceptible tea plant models. The beneficial effects of this invention are: clarifying the role of tea plants in regulating anthracnose resistance / susceptibility. CsNAC72 The role of a gene in the regulation of anthracnose infection was investigated. This gene exhibited differential expression characteristics at different time points of anthracnose infection in tea trees, and could respond to the time-specific induction of anthracnose infection. This clarified its biological function in mediating anthracnose resistance in tea trees, and provided key gene resource support for molecular breeding of tea trees for disease resistance.

Description

Technical Field

[0001] This invention relates to the fields of genetic engineering and plant breeding technology, specifically to tea trees. CsNAC72 Genes, proteins, and their applications in breeding anthrax-resistant varieties and regulating anthrax susceptibility. Background Technology

[0002] Tea is one of my country's most important economic woody crops. Tea leaves contain various substances beneficial to human health, such as flavonoids, tea polyphenols, theanine, and alkaloids. Tea trees thrive in humid and warm environments, which provide favorable conditions for the occurrence of anthracnose. Anthracnose is a fungal disease that is prevalent in late spring, early summer, and autumn, primarily affecting the leaves and young shoots of tea trees, leading to leaf drop, weakened trees, reduced tea yield, and decreased tea quality. Currently, research on tea anthracnose largely focuses on differential gene expression analysis based on transcriptomics, while the discovery of defense-related genes and their biological functions in mediating disease resistance in tea trees are less understood, and their application in disease resistance is severely lacking.

[0003] Chinese patent application CN118440954A discloses a tea tree CsLAC23 gene and its application in resisting anthracnose infection. This patent proposes the application of the tea tree CsLAC23 gene in regulating anthracnose infection in tea trees and in breeding new anthracnose-resistant tea varieties. It proposes an antisense oligonucleotide to inhibit CsLAC23 gene expression. After inhibiting CsLAC23 gene expression and infecting the tea trees with anthracnose, tissue staining and reactive oxygen species (ROS) content measurements showed that the tea leaves with inhibited CsLAC23 gene expression exhibited significantly better DAB and NBT staining effects compared to the control group, and the activities of key ROS scavenging enzymes POD and SOD were lower than those in the control group. Inhibiting CsLAC23 gene expression made the tea trees more sensitive to anthracnose infection, indicating that CsLAC23 enhances the tea tree's resistance to anthracnose stress. However, whether other tea tree genes also have anti-anthrax infection effects has not yet been reported. Summary of the Invention

[0004] The technical problem to be solved by this invention is how to propose a new tea tree gene that can resist anthrax and apply it to resist anthrax infection.

[0005] The present invention solves the above-mentioned technical problems through the following technical means:

[0006] This invention proposes a tea tree CsNAC72 The gene, whose CDS region nucleotide sequence is shown in SEQ ID NO.1.

[0007] The present invention also proposes the above-mentioned tea tree CsNAC72 The protein encoded by the gene has the amino acid sequence shown in SEQ ID NO.2.

[0008] The present invention also proposes the above-mentioned tea tree CsNAC72 Application of genes or proteins in regulating anthracnose resistance in tea trees.

[0009] Preferred, by promoting CsNAC72 Gene expression is used to enhance tea plants' resistance to anthracnose; by inhibiting... CsNAC72 Gene expression is used to reduce the tea plant's resistance to anthracnose.

[0010] The present invention also proposes the above-mentioned tea tree CsNAC72 Application of genes or proteins in the breeding of anthracnose-resistant tea varieties.

[0011] The present invention also proposes the above-mentioned tea tree CsNAC72 Application of genes or proteins in regulating the susceptibility of tea trees to anthracnose.

[0012] Preferably, by inhibiting CsNAC72 Gene expression is used to enhance the susceptibility of tea plants to anthracnose; by promoting... CsNAC72 Gene expression is used to reduce the susceptibility of tea plants to anthracnose.

[0013] This invention also proposes a model of anthracnose-susceptible tea trees, which contains inhibitory agents. CsNAC72 Products that express genes.

[0014] Preferably, the product comprises a VIGS recombinant viral vector carrying the sequence shown in SEQ ID NO.3.

[0015] The present invention also proposes a VIGS recombinant viral vector, which uses the VIGS vector as the base vector and carries the sequence shown in SEQ ID NO.3.

[0016] This invention also proposes a tea plant model for resistance to anthracnose, which contains a tea plant overexpressing the herb *Tea styrax*. CsNAC72 Gene products.

[0017] This invention also proposes a tea tree expression vector pCAMBIA1305- CsNAC72 It was obtained by digesting the fragment shown in SEQ ID NO: 1 into the pCAMBIA1305 vector.

[0018] This invention also proposes a method for cultivating anthracnose-resistant plant varieties, using tea trees... CsNAC72 Genes are introduced into target plants to obtain transgenic plants with enhanced resistance to anthracnose.

[0019] Preferably, it includes the following steps: (1) Cloning tea trees CsNAC72 Gene; (2) Constructing tea trees CsNAC72 Gene overexpression vectors; (3) Tea tree CsNAC72 Transgenic plants with enhanced anthracnose resistance were obtained by transforming the target plants with gene overexpression vectors.

[0020] Preferably, the plant includes one or more of tea trees and tobacco.

[0021] The beneficial effects of this invention are as follows: 1. This invention proposes a tea tree CsNAC72 The role of the gene in regulating anthracnose infection in tea trees was investigated. The expression level of the gene varied at different time points of anthracnose infection in tea trees, and it could respond to the pathogen infection at different time points. This study verified the biological function of the gene in mediating anthracnose resistance in tea trees and provided key gene resources for disease-resistant tea breeding.

[0022] 2. This invention inhibits [the growth of tea plants] within the plant. CsNAC72 Gene expression and infection with anthrax bacteria revealed that the lesion area was significantly larger than that of the control group; the key enzymes POD and SOD for ROS scavenging activity were significantly lower than those in the control group, while the H2O2 content was significantly increased.

[0023] 3. This invention utilizes transient overexpression within the tea plant. CsNAC72 After infection with anthracnose, the necrotic area of ​​the leaves was significantly smaller than that of the control; the content of H2O2, the main component of ROS, was significantly reduced; and the activities of POD and SOD, the key enzymes for scavenging ROS, were significantly higher than those of the control group.

[0024] Of course, implementing any product or method of the present invention does not necessarily require achieving all of the advantages described above at the same time. Attached Figure Description

[0025] Figure 1 The tea tree in Embodiment 1 of the present invention CsNAC72 Schematic diagram of gene expression patterns at different time points during anthrax infection; Figure 2 The tea tree in Embodiment 1 of the present invention CsNAC72 Schematic diagrams of gene expression patterns and transcriptional activity in different tissues; Figure 3 The tea tree in Embodiment 1 of the present invention CsNAC72 Subcellular localization of proteins in tobacco; Figure 4 In Embodiment 1 of the present invention CsNAC72 The results of detecting leaf phenotype, lesion area, and antioxidant enzyme activity in tea plants with transient gene overexpression after infection with anthracnose are shown in the following sub-figures: a represents the results after overexpression. CsNAC72b shows the increase in expression levels; c shows the leaf phenotype under natural light conditions and the leaf phenotype under chlorophyll fluorescence imaging conditions; df shows the statistical results of lesion area after overexpression treatment; and df show the results of the determination of peroxidase (POD) activity, superoxide dismutase (SOD) activity and hydrogen peroxide (H2O2) content in tobacco leaves of the treatment group and the control group, respectively.

[0026] Figure 5 This refers to the virus-induced gene silencing (VIGS) mediated in Example 1 of the present invention. CsNAC72 The effect of gene silencing on anthracnose resistance in tea trees is shown in the figure below: a) shows the results of real-time quantitative PCR analysis, indicating the effect of gene silencing on anthracnose resistance in tea leaves after transient silencing. CsNAC72 Changes in gene expression levels; b represents the treatment group (PTRV2- CsNAC72 Leaf phenotypes and chlorophyll fluorescence imaging of the control group (PTRV2 unloaded) and the treatment group (PTRV2-) under anthracnose infection and non-infection conditions; c shows the leaf phenotypes of the control group (PTRV2 unloaded) and the treatment group (PTRV2-). CsNAC72 The statistical results of the average lesion area after tea leaves were inoculated with anthracnose (n=20); df respectively CsNAC72 Results of the determination of hydrogen peroxide (H2O2), peroxidase (POD) and superoxide dismutase (SOD) content in anthracnose-infected leaves after transient gene silencing treatment; different letters in the figure indicate significant differences between groups (P<0.05, n=5). Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise defined, the technical terms used below have the same meaning as understood by those skilled in the art.

[0028] Unless otherwise specified, the test materials and reagents used in the following examples are commercially available or prepared by known methods.

[0029] Unless otherwise specified, all techniques or conditions described in the embodiments can be performed in accordance with the techniques or conditions described in the literature in this field or in the product manual. Unless otherwise specified, the quantitative experiments in the following embodiments are all repeated three times or more, and the results are averaged.

[0030] Example 1: 1. Tea tree CsNAC72 Cloning of genes (1) Transfer fresh leaf samples of the 'Shuchazao' tea variety to centrifuge tubes containing steel balls and grind them into powder using a ball mill. Subsequent steps were performed according to the FastPure® Universal Plant Total RNA Isolation Kit (Novizan) instructions to extract RNA.

[0031] (2) Reverse transcription to generate the first strand: According to the PrimeScript II 1st Strand cDNA Synthesis Kit (Takara Biotech, China) instructions, 1 μg of RNA was used as a template, and 1 μL of Random 6mers, 1 μL of dNTP Mixture, and RNase-Free H2O were added to a final volume of 10 μL. The mixture was denatured at 65°C for 5 min and immediately placed on ice for 2 min. Then, 4 μL of 5×PrimerScript buffer, 0.5 μL of RNase Inhibitor, 1 μL of PrimerScript RTase, and ddH2O were added to a final volume of 20 μL. The mixture was incubated at 30°C for 10 min, 42°C for 1 h, 95°C for 5 min, and 70°C for 15 min. An appropriate amount of reverse transcribed cDNA was used for subsequent PCR amplification.

[0032] (3) Using the first strand of cDNA as a template, amplification was performed by PCR. CsNAC72 Gene.

[0033] The upstream primer is 5'-ATGGGAGTTGCAGAAACCG-3' (SEQ ID NO.4). Downstream primer: 5'-TTACTGTCGAAGCCTAAATCC-3' (SEQ ID NO.5).

[0034] The reaction mixture consisted of 25 μL of LA Taq premix, 1 μL each of forward and reverse primers, 1 μL of template, and 9.5 μL of ddH2O. The PCR amplification program was as follows: 94℃ for 3 min, 94℃ for 30 sec, 60℃ for 30 sec, 72℃ for 40 sec, and 72℃ for 10 min, for 30 cycles.

[0035] (4) The PCR product obtained in (3) was recovered using a gel extraction kit and ligated into the pEASY-T1 vector (Promega, Shanghai, China) to obtain the recombinant plasmid (pEASY-T1::CsNAC72). This plasmid was then transformed into E. coli competent cells Trans1-T1 and sent to General Biotech for sequencing to obtain the desired results. CsNAC72 The nucleotide sequence of the gene is shown in SEQ ID NO.1. CsNAC72 The specific amino acid sequence encoded by the gene is shown in SEQ ID NO.2.

[0036] 2. CsNAC72 Gene expression patterns under anthrax infection Two-year-old 'Longjing 43' cuttings with uniform growth and free from pests and diseases were selected and cultured in the artificial climate chamber of Anhui Agricultural University (temperature 25℃, humidity 70%, 16 h light, 8 h dark). Anthracnose mycelial blocks were carefully inoculated onto PDA plates using an inoculation needle and cultured at 28℃ in the dark for 7 to 10 days. Aseptic techniques were used throughout the process. ddH2O was added to the plates, and pathogenic spores were scraped from the plates using a sterile spreader. The spores were filtered through sterile gauze and then transferred to sterile Erlenmeyer flasks. A vortex mixer was used to vortex the spores to ensure even distribution. Cell counting was performed under an optical microscope using a hemocytometer. The spores were diluted with sterile water to prepare a 10-1... 7 CFU / mL spore suspension. Wipe two leaves of tea seedlings with 75% ethanol, then wipe again with ddH2O, and let stand until the moisture on the leaves evaporates. Make quincunx-shaped holes in the leaves with a pin, add 50 μL of spore suspension to each side (sterile water was used for the control group). Then wrap the leaves with plastic wrap, cover the entire tea seedling with a large plastic bag, and place it in an artificial climate chamber to maintain humidity above 70%. Samples were taken at 1, 4, 7, 10, and 13 days and immediately flash-frozen in liquid nitrogen, stored at -80℃ for later use.

[0037] RNA was extracted from the samples and reverse transcribed into cDNA. The reverse transcription product was diluted 9-fold and used as a template. A 10 μL reaction mixture was prepared using 2×AceQ Universal qPCR SYBR® Master Mix (Vazyme, Nanjing, China): 5 μL 2×AceQ Universal qPCR SYBR® Master Mix, 1.2 μL diluted reverse transcription product, 0.3 μL each of forward and reverse primers, and 3.2 μL ddH2O. Each sample was tested in triplicate (biological replicates) and triplicate (technical replicates). Quantitative analysis was then performed on a Bio-rad CFX instrument using the following program: 95℃ for 3 min, 95℃ for 10 sec, 56℃ for 30 sec, 65℃ for 5 sec, and 95℃ for 5 sec, for 39 cycles. qRT-PCR was then used for detection. CsNAC72 The changes in gene expression at different time points of anthrax infection, compared with the control group (CK), were observed in the anthrax infection treatment group (Cg). CsNAC72 Gene expression levels significantly increased at 1, 4, 7, 10, and 13 days, with the most significant increase observed in the treatment group at 7 days (e.g., ...). Figure 1 (As shown).

[0038] 3. CsNAC72 Gene expression patterns and subcellular localization in different tissues of tea plants (1) Tea tree CsNAC72 Gene expression patterns in different tissues of tea plants Different tissues, including buds, one-leaf, two-leaf, three-leaf, mature leaves, stems, flowers, and roots, were taken from the national-level superior tea variety 'Shuchazao'. Total RNA and first-strand cDNA were extracted from eight organs for synthesis. Quantitative results showed that... CsNAC72 The gene is expressed in all eight organs, with the highest expression level in the flower, followed by the fruit and root, while the expression level is relatively low in the stem and leaf (e.g., Figure 2 (As shown).

[0039] The upstream primer is 5'-CTAACGAGGGACGAAGAG-3' (SEQ ID NO.6). Downstream primer 5'-TCATCCAACTTTGCGCTTCCAT-3' (SEQ ID NO.7).

[0040] (2) Subcellular localization of CsNAC72 protein in tea plant ① Constructing the target gene expression vector The pCAMBIA1305 vector was double-digested with SpeⅠ and BamHI restriction endonucleases, reacted at 37℃ for 30 min, and then subjected to agarose gel electrophoresis and gel recovery to obtain a linearized vector with restriction sites. Using the target gene plasmid as a template, the plasmid was amplified with primers containing restriction sites to obtain the target gene fragment with restriction sites. The upstream primers were: 5'-GACAGCCCAGATCACTAGTATGGGAGTTGCAGAAAACCG-3' (SEQ ID NO.8), The downstream primer 5'-CTTGCTCACCATGGATCC-3' (SEQ ID NO. 9) was used. The product containing the adapter sequence with the restriction site was recombined with the linearized vector using recombinase. The reaction system was 10 μL: 4 μL of linearized pCAMBIA1305 vector, 2 μL of the product containing the adapter sequence with the restriction site, 1 μL of recombinase E×nase II, and 2 μL of 5×CEII Buffer were added sequentially to a 200 μL PCR tube, and the reaction was carried out at 37℃ for 60 min. The recombinant product was transformed into *E. coli* competent cells Trans1-T1 and sent to General Biosciences for sequencing to obtain pCAMBIA1305- CsNAC72 Plasmid.

[0041] ② Agrobacterium transformation and tea seedling infection Add 1 μL of the recombinant plasmid to EHA105 Agrobacterium competent cells, place on ice for 5 min, incubate with 5 μL of liquid nitrogen at 37°C for 5 min, then place on ice for another 5 min, add 400 μL of liquid LB medium, and then shake on a shaker at 28°C for 2 h. Finally, take 200 μL of the bacterial culture and spread it on cells containing Kansas bacteria. + Incubate on resistant solid LB medium at 28°C in the dark for 48 h. Select single colonies for PCR verification. Add colonies with the verification band position matching the target band position to 400 μL of a solution containing Kansasone propagation agent. + Incubate the culture in resistant liquid LB medium at 28°C with a shaker for 8 hours. Transfer 100 μL of the bacterial culture to a 50 mL container containing Kansas bacteria. + The resistant liquid LB medium was cultured on a shaker at 28°C for 12 h until the OD value reached 0.8–1. The bacterial suspension was centrifuged (5000 r / min, 10 min), the supernatant was discarded, and the cells were resuspended in Agrobacterium resuspension. The OD value of the resuspended bacterial suspension at 600 nm was measured using a UV spectrophotometer to be 0.4–0.6. The suspension was allowed to stand at room temperature for 1 h. The bacterial suspension was then injected into the tobacco leaves using a syringe, ensuring the entire leaf was filled, and the mixture was placed in the dark in an artificial climate chamber for 48 h.

[0042] The infected tobacco leaf was carefully cut off with a blade, and a slide was prepared with the back facing up. The GFP signal was observed under laser confocal microscopy. Green fluorescent protein (GFP) was used for excitation at 488 nm and emission at 509 nm; nuclear localization marker (DAPI); Bright was used for bright-field visualization of cell structure; mCherry was used for membrane localization, with excitation at 587 nm and emission at 610 nm; Merge is a fusion image of GFP, DAPI, Bright, and mCherry. Subcellular localization results showed… CsNAC72 Proteins are located in the cell nucleus (e.g. Figure 3 (As shown).

[0043] 4. Transient expression of CsNAC72 in tea plants to verify its anthracnose resistance function. (1) The pCAMBIA1305-CsNAC72 plasmid was transformed into Agrobacterium, and after successful verification, the culture was shaken. Tea seedlings with uniform growth were selected. The bacterial cultures of pCAMBIA1305 and pCAMBIA1305-CsNAC72 were injected into the tea leaves using a syringe, ensuring that the injection volume of the two bacterial cultures was equal. Quantitative PCR was used to detect the expression of the CsNAC72 gene. The results showed that the expression level of CsNAC72 in the transient overexpression group was significantly higher than that in the empty vector control group (e.g., ...). Figure 4 (as shown in a).

[0044] (2) After 48 h of culture, observe whether expression is observed under a laser confocal microscope. After confirming expression, use a pin to prick quincunx-shaped holes at the injection site, then infect with anthracnose mycelial blocks, wrap with plastic wrap, and use blank culture medium as a control. Observe the phenotype of tea leaves and take pictures after 2 days. The results showed that the lesion area of ​​tea seedlings infected with anthracnose mycelial blocks in the control group was significantly larger than that of tea seedlings overexpressing the protein. Chlorophyll fluorescence showed that the lesion area of ​​tea seedlings after overexpressing CsNAC72 protein was smaller than that of the pCAMBIA-1305 empty vector. Calculation of lesion area showed that the lesion area of ​​tea seedlings after overexpressing CsNAC72 protein was significantly lower than that of the control group (e.g., ...). Figure 4 (As shown in b and c).

[0045] (3) Determination of peroxidase activity and hydrogen peroxide content Tea leaf samples were taken from different sections and immediately flash-frozen in liquid nitrogen. The contents of reactive oxygen species and hydrogen peroxide were then determined according to the method described in section 5 above. Compared to the pCAMBIA-1305 empty vector, tea leaves overexpressing CsNAC72 showed less damage, with significantly increased POD and SOD activities, and significantly decreased H2O2 content (e.g., ...). Figure 4 (As shown in df). This indicates that CsNAC72 mediates the tea plant's resistance to anthracnose infection.

[0046] 5. Virus-induced gene silencing (VIGS) validates the anthrax-resistant function of CsNAC72. (1) Design of VIGS probe The VIGS probe was designed based on the CsNAC72 gene sequence. The specific name and sequence are as follows: PTRV2-CsNAC72-1: 5'-TGAGTAAGGTTACCGAATTCATGGGAGTTGCAGAAACCGAC-3' (SEQ ID NO.3) The designed probe was sent to Shanghai Sangon Biotech Co., Ltd. for synthesis.

[0047] (2) VIGS and anthrax infection ① Select one-year-old 'Longjing 43' cuttings with uniform growth and free from pests and diseases. Inject the diluted probe into two leaves using a 1 mL syringe. Use sterile water as a control. After 14 days, collect the two leaves and freeze them in liquid nitrogen to extract RNA. Use the cDNA obtained from reverse transcription as a template for qPCR verification to screen the best probe (SEQ ID NO.3). The results showed that after 14 days of silencing, the expression level of CsNAC72 was significantly lower than that of the water control group (e.g., ...). Figure 5 (as shown in a).

[0048] ② Tea seedlings were treated with the selected optimal VIGS probe, with PTRV2 empty vector used as a control. After 14 days, tea leaves were infected with anthracnose spore suspension. Leaves infected for 3 days and leaves from the control group were taken for phenotypic observation, and leaf phenotypes were observed using an IMAGE-PAM modulated fluorescence spectrometer (WALZ, Germany).

[0049] (3) Phenotypic statistics and physiological index determination of anthracnose-infected tea leaves after CsNAC72 gene silencing ① The area of ​​lesions after VIGS inhibition was statistically analyzed, and ImageJ software was used for area calculation. The sample size for each treatment was 20 tea leaves. Results showed that the VIGS inhibition group (PTRV2- CsNAC72 The leaf lesion area was significantly larger than that of the control group (PTRV2) (e.g.) Figure 5 (As shown in b and c).

[0050] ② Determination of hydrogen peroxide content and peroxidase activity The H2O2 content, POD, and SOD enzyme activities of tea leaves infected with VIGS for 14 days were determined. The specific steps are as follows: Total protein was extracted from the sample; 0.06 g of the sample was weighed and ground into powder using liquid nitrogen; then transferred to a pre-chilled mortar with 540 μL of 10×PBS; and ground into a homogenate on ice; centrifuged at 12,000 g for 10 min; the supernatant was collected for analysis; based on H2O... 2、The instructions for the POD and SOD test kits (Nanjing Jiancheng) indicate that H2O2 content, POD, and SOD enzyme activity were measured respectively. After anthracnose fungus infected tea leaves, compared to PTRV2 treatment, inhibiting CsNAC72 gene expression significantly increased H2O2 content. Figure 5 d), both POD and SOD enzyme activities decreased significantly (e.g. Figure 5 (As shown in the figure). These results indicate that the CsNAC72 gene enhances the tea plant's resistance to anthracnose by positively regulating the antioxidant enzyme system and maintaining reactive oxygen species (ROS) metabolic balance. Silencing CsNAC72 not only reduces the activity of antioxidant enzymes but also exacerbates the accumulation of ROS, leading to increased oxidative stress and ultimately weakening the tea plant's disease resistance.

[0051] The nucleotide sequence shown in SEQ ID NO.1 of this invention is specifically as follows: The amino acid sequence shown in SEQ ID NO.2 of this invention is specifically as follows: MGVAETDLLSQLRLPPGFRFYPTDEELLVQYLCRKVAGYHFSLQIIAEIDLYKFDPWVLPSKAIFGEKEWYFFSPRDRKYPNGSRPNRVAGSGYWKATGTDKIITNEGRRVGIKKALVFYVGKAPKGTKTNWIMHEYRLSEPPKKNGSAKLDDWVLCRIYKKNSSAQKPISGVS SKELSHGSSSSSSSQFDDMLESLPEINDRFFTLPRMNSLKTIQQDEKFNIQNLTSGNFDWASLAGAVTSLPELVPGGQSQAQTQAQGHLGNNNNMYNDMYVPSIPPLSRVDSPCERFGASVEEEVQSGLRTQHRFENSGFFQQNSNGFAQSFSNSPDPFGIRYPTQQGGFRLRQ The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A type of tea tree CsNAC72 Genes, characterized by, The nucleotide sequence of its CDS region is shown in SEQ ID NO.

1.

2. The tea tree as described in claim 1 CsNAC72 Gene-encoded proteins are characterized by, Its amino acid sequence is shown in SEQ ID NO.

2.

3. The tea tree as described in claim 1 CsNAC72 The application of the gene or the protein described in claim 2 in regulating anthracnose resistance / susceptibility in tea trees.

4. The tea tree as described in claim 1 CsNAC72 The application of the gene or the protein described in claim 2 in the breeding of anthracnose-resistant tea varieties.

5. A model of anthracnose-susceptible tea trees, characterized in that, This model contains methods to inhibit the tea tree as described in claim 1. CsNAC72 Products that express genes.

6. The anthracnose-sensitive tea tree model according to claim 5, characterized in that, The product includes a VIGS recombinant viral vector carrying the sequence shown in SEQ ID NO.

3.

7. A tobacco model resistant to anthrax, characterized in that, The model contains the overexpression of the tea plant of claim 1 in tobacco. CsNAC72 Gene products.

8. A tea tree expression vector pCAMBIA1305- CsNAC72 Its characteristics are, It was obtained by digesting the fragment shown in SEQ ID NO: 1 into the pCAMBIA1305 vector.

9. A method for cultivating anthracnose-resistant plant varieties, characterized in that, The tea tree as described in claim 1 CsNAC72 Genes are introduced into target plants to obtain transgenic plants with enhanced resistance to anthracnose.

10. The cultivation method according to claim 9, characterized in that, Specifically, the following steps are included: (1) Cloning tea trees CsNAC72 Gene; (2) Constructing tea trees CsNAC72 Gene overexpression vectors; (3) Tea tree CsNAC72 Transgenic plants with enhanced anthracnose resistance were obtained by transforming the target plant with a gene overexpression vector; the plant was a tea tree.

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