Camellia sinensis gene CsGSII-1b and application thereof in improving plant cold tolerance
By cloning and regulating the CsGSII-1b gene in tea trees, the problem of tea trees being susceptible to frost damage at low temperatures has been solved, the cold resistance of tea trees has been improved, new ideas and practical references have been provided for tea tree breeding, and the sustainable development of the tea industry has been promoted.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- ANHUI AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-03-16
- Publication Date
- 2026-06-05
AI Technical Summary
Tea trees are susceptible to frost damage under low temperature conditions, resulting in losses in yield and quality. Current technology lacks effective methods to improve cold resistance.
The CsGSII-1b gene of tea plant was cloned and verified. The low temperature tolerance of tea plant was regulated by overexpressing or inhibiting the CsGSII-1b gene in tea plant through genetic engineering.
It improved the low-temperature tolerance of tea trees, reduced damage under low-temperature stress, promoted the breeding of resistance traits in tea trees, and provided theoretical and practical references for the sustainable development of the tea industry.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of genetic engineering technology, specifically to the CsGSII-1b gene in tea trees and its application in improving plant cold resistance. Background Technology
[0002] The tea plant (Camellia sinensis (L.) O. Kuntze) is an important economic crop primarily harvested for its tender buds, and it thrives in warm climates but is sensitive to cold. Tea leaves can be processed into beverages and contain various beneficial components with health benefits. Currently, high-quality green tea has become a core driver of the consumer market and holds a dominant position. However, in recent years, tea trees have frequently suffered from low-temperature damage such as frost or late spring cold snaps during the winter and early spring. Damage to the new shoots of the tea trees has resulted in significant losses in the yield and quality of spring tea, sometimes even leading to total crop failure. Low-temperature freezing damage has become one of the most significant and serious natural disasters in tea production, threatening the healthy and sustainable development of the tea industry.
[0003] Glutamine synthetase (GS) was the first enzyme isolated, purified, and identified from plants. It is widely distributed in all organisms and was also the first enzyme discovered to catalyze the assimilation of ammonia from inorganic salts into organic nitrogen forms. Specifically, it utilizes the energy released from the hydrolysis of ATP to ADP to catalyze the synthesis of glutamine from ammonia and glutamate, playing a crucial role in the nitrogen metabolism pathway (GS / GOGAT cycle) of organisms. Nitrogen is an essential element for plant growth and development, playing a vital role in crop yield and quality. In recent years, increasing research has revealed that plant GS can respond to various abiotic stresses. Therefore, exploring the biological function of glutamine synthetase in regulating the cold resistance of tea trees is beneficial for understanding the complex molecular mechanisms of tea tree cold resistance, providing new insights into the cold resistance mechanism of tea trees, and offering important theoretical basis for breeding tea trees with resistance traits. Summary of the Invention
[0004] The purpose of this invention is to provide the CsGSII-1b gene of tea tree and its application in improving the cold resistance of plants. It provides a new idea for the cold resistance mechanism of tea tree and provides a theoretical and practical reference basis for the breeding of tea tree resistance traits.
[0005] In one aspect of the present invention, the present invention proposes a tea plant CsGSII-1b gene. According to an embodiment of the present invention, the tea plant CsGSII-1b gene is a tea plant glutamine synthase gene, and the nucleotide sequence of the tea plant CsGSII-1b gene is shown in SEQ ID NO.1 of the sequence listing.
[0006] In another aspect of the invention, the present invention proposes an application of the CsGSII-1b gene in improving plant cold tolerance. According to an embodiment of the invention, inhibition of CsGSII-1b gene expression in tea plants can improve the plant's low-temperature tolerance.
[0007] Compared with the prior art, the beneficial effects of the present invention are:
[0008] This invention marks the first cloning and verification of the function of the glutamine synthase gene GSII-1b (i.e., CsGSII-1b) in regulating the formation of cold resistance in tea plants. This invention also provides a recombinant plasmid containing the CsGSII-1b gene and a transgenic engineered bacterium. This invention enriches the research on glutamine synthase in tea plants, provides new insights into the mechanism of cold resistance in tea plants, and offers a theoretical and practical reference basis for breeding tea plants with resistance traits. Attached Figure Description
[0009] Figure 1 In the diagram, A and B represent the expression pattern of the tea tree gene CsGSII-1b under 4℃ low-temperature treatment and the expression status in different tissues, respectively, in Example 2 of this invention.
[0010] Figure 2 In the diagram, A, B, C, and D represent the expression analysis of gene CsGSII-1b overexpression in tea leaves, the phenotypic analysis of overexpressed leaves under low-temperature treatment, the Fv / Fm value, and the malondialdehyde content analysis, respectively, in Example 3 of the present invention; E, F, G, and H represent the chlorophyll fluorescence of damaged plants with antisense oligonucleotide inhibition of CsGSII-1b, the expression level change, the Fv / Fm value, and the malondialdehyde content analysis, respectively, in Example 3 of the present invention.
[0011] Figure 3 In the diagram, A, B, and C represent the verification diagram of Arabidopsis thaliana lines overexpressing gene CsGSII-1b in Example 4 of this invention, the phenotypic analysis diagram of the overexpressing lines under low temperature stress, and the quantitative diagram of chlorophyll fluorescence Fv / Fm value, respectively. Detailed Implementation
[0012] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.
[0013] Unless otherwise stated, the terms used in this invention generally have the meanings commonly understood by those skilled in the art.
[0014] The present invention will be described in further detail below with reference to specific preparation and application examples and data. It should be understood that these examples are merely illustrative of the invention and are not intended to limit the scope of the invention in any way.
[0015] In the following embodiments, the various processes and methods not described in detail are conventional methods known in the art. All primers used are indicated upon their first appearance, and subsequent use of the same primers is identical to their initial indication.
[0016] Example 1: Cloning and Sequence Structure Analysis of the CsGSII-1b Gene
[0017] CsGSII-1b is the glutamine synthase gene from tea plants. Its cloning and sequence structure analysis are as follows:
[0018] The tea plant material used in this experiment was the national-level superior tea variety "Shuchazao," planted in the Agricultural Industrial Park of Anhui Agricultural University. Tender leaves were used for RNA extraction. Total RNA was extracted using the RNAprep Pure Plant Kit (Tiangen, Beijing, China) according to the instructions. The RNA content and quality were detected using a spectrophotometer.
[0019] First-strand reverse transcription: Using 1 μg of RNA as a template, prepare the following according to the PrimeScript II 1st Strand cDNASynthesis Kit (Takara Biotech, China) instructions: add 0.6 μl of Oligo dT Primer (50 μM), 0.4 μl of Random 6 mers (50 μM), 1 μl of dNTP Mixture (10 mM each), and bring the RNase-free dH2O to a final volume of 10 μl. Denature at 65°C for 5 min and immediately place on ice. Then add 4 μl of 5×PrimerScript buffer, 0.5 μl of RNase Inhibitor (40 U), 1 μl of PrimeScript RTase (200 U), and bring the dH2O to a final volume of 20 μl. Incubate at 42°C for 45 min, then at 95°C for 5 min to inactivate the reverse transcriptase. After optimization, use an appropriate amount of the reverse transcription product for subsequent PCR. The gene sequence was amplified by PCR using cDNA first strand as an RT-PCR template and conventional methods.
[0020] The upstream primer is: (5'-ATGTCTCTTCTTTCCGATCTTTGC-3'),
[0021] The downstream primer is: (5'- TTACGGTTTCCAGAGGATGGTG -3').
[0022] The 50 μl PCR reaction system consisted of: 25 μl of 2×Phanta Max buffer, 1.0 μl of dNTP Mix, 2 μl each of forward and reverse primers, 1 μl of Phanta Max Supper-Fidelity DNA Polymerase, 2 μl of template, and 17 μl of ddH2O.
[0023] The PCR reaction program was as follows: 95℃ for 3 min, 95℃ for 15 sec, 59℃ for 15 sec, 72℃ for 30 sec, 35 cycles, 72℃ for 5 min, and 4℃ to finish. The PCR product, CsGSII-1b gene, was purified and recovered, then ligated into the PGEX4T-1 vector (Biofeng, Shanghai, China) to obtain the PGEX4T-1::CsGSII-1b plasmid. This plasmid was transformed into *E. coli* competent cells DH5α (WEIDI, Shanghai, China). After positive clones grew and were verified by colony PCR, single clones that were verified were selected for sequencing. The nucleotide sequence of the CsGSII-1b gene is shown in SEQ ID NO.1 of the sequence listing, as follows:
[0024]
[0025] The protein sequence encoded by the CsGSII-1b gene is shown in SEQ ID NO.2, and is as follows:
[0026] MSLLSDLCNLNLSESTEKIIAEYIWIGGSGMDLRSKARTLNAPVSDPSKLPQWNYDGSSTGQAPGEDSEVILYPQAIYKDPFRRGNNILVMCDAYTPGGEPIPTNKRFDAAKIFSHPDVVAEEPWYGIEQEYTLLQKEVKWPIGWPVGGYPGPQGPYYCGIGADKAFGRDIVDAHYKA CLYAGINISGINGEVMPGQWEFQVGPSVGISSGDQLWMARYILERITEIAGVVVSFDPKPIEGDWNGAGAHTNYSTKSMRSDGGFEVIKKAIEKLGLKHKEHIAAYGEGNERRLTGKHETADINTFLWGVANRGASIRVGRDTEKAGKGYFEDRRPASNMDPYVVTSMIANTTILWKP.
[0027] Example 2: Differential expression analysis of the CsGSII-1b gene
[0028] (1) Expression of CsGSII-1b gene in different tissues of tea plant
[0029] Seven tissues from the national-level superior tea variety Shuchazao were used to analyze gene expression. These seven tissues included buds, one-leaf, two-leaf, three-leaf, four-leaf, young stems, and roots. These samples were also used for total RNA extraction and first-strand cDNA synthesis. The reverse transcription product (cDNA first strand) was diluted 30-fold as a template and synthesized using Hieff. TM qPCR SYBR ® GreenMaster Mix (No Rox) (Yeasen, Shanghai, China) was used to prepare a 20 μl reaction system: 2.0 μl of reverse transcription product diluted 30-fold, 0.4 μl each of forward and reverse primers (10 pmol / μl), and 10 μl of Hieff. TM qPCR SYBR ®Green MasterMix, 7.2 μl ddH2O, 3 replicates per reaction. Then, on a Bio-rad CFX-96 instrument, the following program was used: ① 95℃ for 5 min; ② 39 cycles of 95℃ for 10 sec, 60℃ for 30 sec, and 72℃ for 30 sec; ③ Melting curves were plotted at 0.1°C / sec from 65℃ to 95℃. Upstream primer: (5'-AAGTGAAGTGGCCGATTGGT-3'), downstream primer: (5'-TTGGAATTCCCACTGACCCG-3'). Using the tea plant ACTIN gene as an internal control, upstream primer: (5'-GCCATATTTGATTGGAATGG-3'), downstream primer: (5'-GGTGCCACAACCTTGATCTT-3'). The relative expression levels of CsGSII-1b in different tissues were calculated using the instrument's built-in analysis software.
[0030] (2) Analysis of the expression pattern of CsGSII-1b under low temperature stress
[0031] Tea tree branches of uniform growth were selected for the experiment and placed in a 4℃ incubator. A control group was placed at room temperature. The first or second tender leaves of both control and treated plants were collected at different time points (0d, 1d, and 3d) in triplicate. The leaves were then frozen in liquid nitrogen at -80℃ for analysis of CsGSII-1b gene expression. RNA extraction and quantitative PCR were performed using the same methods.
[0032] Figure 1 This study examines the expression changes of CsGSII-1b under 4℃ low-temperature treatment and the expression differences in different tissues of tea plants. Figure 1 As shown, using the tea cultivar Shuchazao as the research material, CsGSII-1b exhibited a downregulated expression pattern without low-temperature treatment, and the expression pattern in different tissues showed tissue specificity. Figure 1 The qPCR results showed that CsGSII-1b expression was negatively correlated with the tea plant's response to low-temperature stress. CsGSII-1b was expressed in a constitutive manner in all tissues. This suggests that CsGSII-1b may be closely related to the tea plant's response to low-temperature stress.
[0033] Example 3: Functional verification of the CsGSII-1b gene in tea plants
[0034] 3.1 Transient overexpression experiment in vivo
[0035] (1) Construction of CsGSII-1b-PCAMBIA1305 vector
[0036] Using the cDNA plasmid as a template, PCR amplification was performed with the upstream primer (5'-CAGATCACTAGTATGTCTCTTCTTTCCGATCTTTGC-3') and the downstream primer (5'-CACATGGATCCTTACGGTTTCCAGAGGATGGTG-3'). The PCR products were analyzed by 1% agarose gel electrophoresis to confirm specificity, followed by purification and elution. The PCAMBIA1305 vector was linearized by double enzyme digestion, using Spe1 and BamH1 as digestion sites. The digestion system consisted of: 41.4 μl ddH2O, 5 μl 10ⅹK Buffer, 1.6 μl PCAMBIA1305_GFP Vector, 1 μl Spe1, and 1 μl BamH1, incubated at 37°C for 2 h. The digestion products were analyzed by 1% agarose gel electrophoresis to confirm cleavage, followed by purification and elution. Recombinant reactions were performed using a recombinase (GenRec Assembly MasterMix Kit, GENERAL BIOL) to digest the linearized PCAMBIA1305 vector and purified PCR product. The reaction mixture consisted of 5 μl of 2×GenRec Assembly Master Mix, 1.5 μl of PCR product, and 3.5 μl of vector, incubated at 50 °C for 40 min. The resulting product was then transformed into competent E. coli DH5α cells (WEIDI, Shanghai, China). After positive clones emerged and were verified by colony PCR, single clones that were verified were selected for sequencing to obtain the CsGSII-1b-PCAMBIA1305 vector.
[0037] (2) Agrobacterium injection permeation method for tea leaves
[0038] The CsGSII-1b-PCAMBIA1305 vector was transformed into Agrobacterium GV3101 using the freeze-thaw method, and positive clones were identified by colony PCR. Single clones verified by colony PCR were picked and inoculated into 3 ml of liquid LB medium (containing 50 μg / ml Rifampicin). ＋ and 50mg / ml Kan ＋ Incubate at 28℃ and 200 rpm for 16-18 hours until OD reaches its maximum. 600 =0.8-1.2. Inoculate 3 ml of overnight cultured Agrobacterium into 50 ml of liquid LB medium containing (50 μg / ml rifampicin). ＋ and 50mg / ml Kan ＋ Incubate at 28℃ and 200 rpm for 6-8 hours, collect bacterial cells by centrifugation, and treat with 10 mM MgCl2 and 10 mM... 2- The bacterial cells were resuspended in a resuspending solution with (N-morpholino)ethanesulfonic acid adjusted to pH 5.6 until OD was reached.600 =0.9-1. Add 1 / 1000 100μM acetylsylgenone (As) to the bacterial solution and incubate in the dark at 28℃ for 2 hours. Select healthy early-maturing tea seedlings of the same growth period, make a small incision on the lower epidermis of the leaves 1-2 cm from the leaf tip or petiole, and inject the bacterial solution using a disposable 1ml syringe (without the needle) to allow the solution to penetrate the entire leaf tissue. After injection, treat the seedlings in the dark at room temperature for 24 hours, then resume normal culture for 24 hours. Quickly fix the injected leaves with liquid nitrogen and store them at -80℃.
[0039] (3) Low-temperature treatment and biochemical index determination of overexpression samples
[0040] To investigate the role of CsGSII-1b in the response of tea plants to low-temperature stress, CsGSII-1b overexpression samples were treated at -2℃ for 2 h, then allowed to recover to room temperature for 30 min, and changes in chlorophyll fluorescence intensity (FV / FM) were observed. Subsequently, the malondialdehyde (MDA) content of the low-temperature treated samples was determined according to the kit (catalog number: BC0020) (Solarbio, Beijing, China).
[0041] 3.2 In vivo antisense oligonucleotide inhibition experiment
[0042] (1) Antisense oligonucleotide primer design
[0043] Oligonucleotide primers were designed based on the coding sequence of the CsGSII-1b clone. The design was completed on the website http: / / sfold.wadsworth.org / cgi-bin / soligo.pl, and the primer sequences are shown below:
[0044] AsODN-1, (5' - GGAAGTGACAACGTACGGATCCATGTTTG -3');
[0045] AsODN-2, (5' - TTTGCCTGCCTTCTCTGTGTCCCGGCCAA -3');
[0046] AsODN-3, (5' - TCCAGTCACCCTCAATAGGTTTAGGGTCG -3');
[0047] AsODN-4, (5' - GGTGCATTCAGGGTCCTGGCTTTGCTTCT -3');
[0048] Primers were dissolved in sterile water to prepare oligonucleotide primer solutions, with ddH2O as a control. New tea shoots of the same growth period and in good condition were selected, and their bottom ends were immersed in 1.5 ml centrifuge tubes containing 1 ml of 20 μM primer solution. The centrifuge tubes were then placed in a room temperature incubator for 24 hours of normal incubation. The shoots were then rapidly fixed with liquid nitrogen and stored at -80°C.
[0049] (2) Analysis of antisense oligonucleotide inhibition of CsGSII-1b expression
[0050] Total RNA was extracted from both the treated and control samples, and first-strand cDNA was synthesized via reverse transcription. The expression of the CsGSII-1b gene was detected using qPCR. The results showed that CsGSII-1b expression was significantly inhibited.
[0051] (3) Low-temperature treatment and biochemical index determination of antisense oligonucleotide inhibitory samples
[0052] To investigate the role of CsGSII-1b in the response of tea plants to low-temperature stress, samples with inhibited CsGSII-1b expression were treated at -2℃ for 1 h, then allowed to recover to room temperature for 30 min, and changes in chlorophyll fluorescence intensity (FV / FM) were observed. The malondialdehyde (MDA) content of the subsequently low-temperature treated samples was determined according to the kit (catalog number: BC0020) (Solarbio, Beijing, China). Approximately 0.1 g of freshly ground sample powder was added to 1 ml of extraction buffer and homogenized in an ice bath. The mixture was centrifuged at 8000 g at 4℃ for 10 min, and the supernatant was placed on ice for testing. In the experimental group, 600 μl of MDA detection working solution, 200 μl of the test sample, and 200 μl of reagent three were added sequentially. In the control group, 600 μl of MDA detection working solution, 200 μl of distilled water, and 200 μl of reagent three were added sequentially. The mixture was incubated in a 100℃ water bath for 60 min, then cooled in an ice bath and centrifuged at 10000 g at room temperature for 10 min. Transfer the supernatant to a 1 ml glass cuvette and measure the absorbance of each sample at 450 nm, 532 nm, and 600 nm. Calculate ΔA450 = A450 for each sample. 测定 -A450 空白 ΔA532=A532 测定 -A532 空白 ΔA600=A600 测定 -A600 空白 MDA content (nmol / g) = 5 * [6.45 * (ΔA532 - ΔA600) - 1.29 * ΔA450] / W.
[0053] like Figure 2As shown, overexpression of CsGSII-1b significantly exacerbated low-temperature damage to tea leaves, leading to a significant decrease in the Fv / Fm ratio and a significant increase in malondialdehyde (MDA) content. In the antisense oligonucleotide inhibition experiment, CsGSII-1b expression was significantly suppressed compared to the control. After low-temperature treatment, compared to the control group, plants with suppressed CsGSII-1b expression showed less damage, a significantly increased Fv / Fm ratio, and a significantly decreased MDA content. These results indicate that the CsGSII-1b gene can negatively regulate the low-temperature tolerance of tea plants.
[0054] Example 4: Functional validation of CsGSII-1b overexpression in Arabidopsis thaliana
[0055] (1) Genetic transformation in Arabidopsis thaliana
[0056] The CsGSII-1b-PCAMBIA1305 vector was constructed as above. An appropriate amount of wild-type Arabidopsis seeds were added to deionized water and vernalized for 72 hours before sowing. After sowing, the seeds were covered with plastic wrap and placed under suitable conditions (60% humidity; 23℃; 16h light / 8h dark photoperiod) to await germination. After germination, seedlings of uniform size were selected for transplanting and normal culture. The CsGSII-1b-PCAMBIA1305 vector was transformed into Agrobacterium GV3101 using a freeze-thaw method, and positive clones were identified by colony PCR. Positive colonies containing the target gene were picked and cultured in 50 mL of LB broth containing the corresponding antibiotic at 28℃ and 200 rpm for approximately 24 hours. 50 mL of the cultured bacterial solution was added to 200 mL of fresh LB broth containing the corresponding antibiotic, and cultured with shaking for 6-8 hours until OD (Organic Dioxide) was reached. 600 The bacterial cells were collected by centrifugation at approximately 1.0, and then resuspended in 5% sucrose solution. The final concentration was OD100. 600 The concentration is approximately 0.8. Add 0.1% silwet L-77 and shake well. After about a month of planting, the Arabidopsis thaliana plants will begin to flower. Select healthy plants as the plants to be transformed. Before transformation, continuously remove the terminal inflorescences to encourage the plants to produce more flower buds. The plants to be transformed should be thoroughly watered the day before transformation.
[0057] The prepared transformation solution was placed in a container, and the Arabidopsis inflorescences were gently immersed in the transformation solution for about 60 seconds. Afterward, the inflorescences were placed in the dark for 24 hours, followed by normal culture, and the seeds were harvested. The harvested Arabidopsis seeds were placed in centrifuge tubes, sterilized with 1 ml of 75% ethanol for 1 min, then sterilized with 10% NaClO for 5 min, and rinsed 5-6 times with sterile water. The seeds were then aspirated with a pipette tip and sown on 1 / 2 MS solid medium containing hygromycin. Vernalization was carried out at 4℃ in the dark for 72 hours, followed by transfer to a culture room at 23℃ with a photoperiod of 16 h light / 8 h dark. After about two weeks, resistant plants with green leaves and normal root development were selected and transplanted into a cultivation substrate for further culture. The cultivation substrate was fully saturated with water before transplanting, and covered with plastic wrap after transplanting. This was removed after about 3 days, and subsequent management was the same as above. T2 generation seeds were harvested for experiments. RNA was extracted from Arabidopsis seedlings, and PCR was performed using gene-specific primers to detect the expression of the target gene. After culturing the transgenic plants at -8℃ for 2.5 hours, the culture dishes were removed and then transferred to a normal culture room for further culture. The survival of the seedlings was observed after 3 days.
[0058] Figure 3 This section describes the expression analysis of CsGSII-1b in Arabidopsis thaliana overexpression lines and wild-type strains, as well as the expression analysis of overexpression lines under low-temperature treatment. Figure 3 As shown, PCR analysis revealed that CsGSII-1b was integrated into Arabidopsis plants during the overexpression experiment. The expression level of the CsGSII-1b gene in transgenic (OE) plants was significantly higher than that in wild-type (WT) plants. Both CsGSII-1b overexpression lines were subjected to -3℃ for 2 hours. Compared to wild-type Arabidopsis, the transgenic lines showed significantly increased damage and a significantly higher FV / FM value, indicating that CsGSII-1b overexpression reduced the plant's tolerance to low temperatures.
[0059] In summary, the expression pattern of CsGSII-1b is associated with the tea plant's response to low-temperature stress. Overexpression of this gene can reduce the plant's cold tolerance, a result verified in CsGSII-1b-overexpressing transgenic Arabidopsis thaliana. Furthermore, inhibiting CsGSII-1b expression using antisense oligonucleotide technology significantly reduced leaf damage at low temperatures in tea plants. These results indicate that the CsGSII-1b gene participates in the tea plant's response to low-temperature stress. Cloning this gene will not only facilitate the investigation of the role of glutamine synthase in the tea plant's cold tolerance process but also contribute to promoting genetic improvement aimed at enhancing tea plant cold tolerance, thus promoting the sustainable development of the tea industry. This invention has significant application value.
[0060] This invention marks the first cloning and verification of the glutamine synthase gene CsGSII-1b, which regulates the tea plant's response to low-temperature stress. This gene plays a crucial regulatory role in the tea plant's response to low-temperature stress. This invention also provides a recombinant plasmid containing the CsGSII-1b gene and a transgenic engineered bacterium. This invention enriches the research on transcription factors in tea plants, provides new insights into the cold resistance mechanism of tea plants, and offers a theoretical and practical reference basis for breeding tea plants with resistance traits.
[0061] The above description is merely an example and illustration of the structure of the present invention. Those skilled in the art can make various modifications or additions to the specific embodiments described, or use similar methods to replace them, as long as they do not deviate from the structure of the present invention or exceed the scope defined in the claims, all of which should fall within the protection scope of the present invention.
Claims
1. The CsGSII-1b gene of tea plant, characterized by: The CsGSII-1b gene is a glutamine synthase gene from tea plants, and the nucleotide sequence of the CsGSII-1b gene from tea plants is shown in SEQ ID NO.
1.
2. The application of the CsGSII-1b gene in tea plants to enhance cold resistance, characterized by: The inhibition of CsGSII-1b gene expression in tea plants according to claim 1 can improve the plant's low-temperature tolerance.