Application of TeLCYE protein and biological material thereof in increasing carotenoid content and salt tolerance of plants

Abstract

The invention belongs to the technical field of biology, and particularly relates to application of TeLCYE protein and a biological material thereof in increasing the carotenoid content and salt tolerance of plants, the TeLCYE protein is derived from marigold, and the TeLCYE protein is a protein with an amino acid sequence shown as SEQ ID NO: 1; the related biological material of the TeLCYE protein comprises a nucleic acid molecule for coding the TeLCYE protein, and the nucleic acid molecule is a DNA molecule with a nucleotide sequence as shown in SEQ ID NO.2; the invention relates to application of TeLCYE protein and related biological materials thereof in improvement of carotenoid content and salt tolerance of plants. Cultivating a transgenic plant capable of improving the carotenoid content and the salt tolerance; the TeLCYE protein and the coding gene thereof provided by the invention have important application values in improvement of the carotenoid content of plants and stress resistance research, and have wide application space and market prospects in the agricultural field.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to application of TeLCYE protein and biological materials thereof in improving the carotenoid content and salt tolerance of plants. Background Art

[0002] Carotenoids are an important class of terpenoid compounds. In plants, they not only participate in the photoprotection mechanism in photosynthesis, but also act as antioxidants to reduce the damage of reactive oxygen to cells. In addition, carotenoids are precursors of vitamin A and are essential for human health. Its synthesis pathway involves multiple key enzyme genes, such as PSY (phytoene synthase), PDS (phytoene dehydrogenase) and LCY (lycopene cyclase). The regulation of these genes directly affects the types and content of carotenoids. Marigold (Tagetes erecta) has become a research hotspot because its flowers are rich in lutein (a type of carotenoid). The carotenoid content in its flowers is as high as 11%-17%, and it has significant antioxidant and coloring functions. It is widely used in feed additives and natural pigment extraction. Studies have shown that the expression of carotenoid metabolism-related genes (such as TeLCY) in mature marigold flowers is significantly upregulated, indicating that its developmental stage-specific regulatory mechanism may provide targets for genetic engineering. TeLCYE belongs to the lycopene cyclase family, catalyzing the conversion of lycopene to ε-carotene and affecting the balance of the carotenoid branch pathway. In marigold, the expression level of this gene is positively correlated with carotenoid accumulation. The accumulation of carotenoids is closely related to plant stress resistance. For example, transgenic plants overexpressing the Lycium barbarum LcLCY gene enhance antioxidant capacity and reduce ROS accumulation under salt stress by increasing carotenoid content, thereby improving stress resistance. The carotenoid metabolism genes of marigold may participate in environmental stress responses such as high temperature, drought or saline-alkali stress through similar mechanisms.

[0003] Salinization is one of the main forms of land degradation, with about 1 billion hectares of land affected by salinization worldwide. Salinization leads to a decline in soil fertility and crop yields, which seriously threatens food security. The high salt and alkalinity of saline-alkali land seriously affect plant growth, leading to a decline in agricultural output. Modifying plants through molecular biology to adapt them to saline-alkali environments is an effective way to solve this problem.

[0004] Therefore, cloning the key gene TeLCYE in marigold carotenoid biosynthesis, using genetic engineering technology to increase the carotenoid content of plants, while improving the salt resistance of plants, and breeding new plant varieties with high carotenoid content are important ways to improve crop quality and production performance. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies in the prior art and provide a TeLCYE protein and a biomaterial thereof for use in improving the carotenoid content and salt tolerance of plants.

[0006] In order to achieve the purpose of the present invention, the present invention is implemented by adopting the following technical solutions.

[0007] A TeLCYE protein, wherein the TeLCYE protein is derived from marigold; the TeLCYE protein is a protein having an amino acid sequence as shown in SEQ ID NO: 1; wherein the amino acid sequence as shown in SEQ ID NO: 1 is:

[0008] .

[0009] As a preferred embodiment of the present invention, the TeLCYE protein can be artificially synthesized, or its encoding gene can be synthesized first and then obtained by biological expression.

[0010] As a preferred embodiment of the present invention, the TeLCYE protein can be expressed, detected, traced and / or purified by in vitro DNA recombination technology using a protein tag.

[0011] As a preferred embodiment of the present invention, the protein tag includes: Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag and / or SUMO tag.

[0012] A TeLCYE protein-related biological material, wherein the related biological material is any one of the following related biological materials:

[0013] (1) A nucleic acid molecule encoding a TeLCYE protein, wherein the nucleic acid molecule is a DNA molecule having a nucleotide sequence as shown in SEQ ID NO.2;

[0014] (2) A recombinant vector containing the nucleic acid molecule described in (1);

[0015] (3) A recombinant microorganism containing the nucleic acid molecule described in (1);

[0016] (4) A transgenic plant cell line containing the nucleic acid molecule described in (1);

[0017] (5) Transgenic plant tissue containing the nucleic acid molecule described in (1);

[0018] (6) Transgenic plant organs containing the nucleic acid molecule described in (1);

[0019] (7) A transgenic plant containing the nucleic acid molecule described in (1);

[0020] (8) Tissue culture produced by regenerable cells of transgenic plants containing the nucleic acid molecule described in (1);

[0021] (9) Protoplasts produced by tissue culture containing the nucleic acid molecule described in (1).

[0022]

[0023] As a preferred embodiment of the present invention, a recombinant vector containing the TeLCYE encoding gene expression cassette can be constructed using a plant expression vector. The plant expression vector can be a Gateway system vector or a binary Agrobacterium vector, such as pGWB411, pGWB412, pGWB405, pBin438, pCAMBIA1300, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1301, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb. When using TeLCYE to construct a recombinant vector, any enhanced, constitutive, tissue-specific or inducible promoter can be added before its transcription initiation nucleotide, such as cauliflower mosaic virus (CAMV) 35S promoter, ubiquitin gene Ubiqutin promoter (pUbi), etc., which can be used alone or in combination with other plant promoters; in addition, when using the gene of the present invention to construct a plant expression vector, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signal and the start codon are extensive and can be natural or synthetic. The translation initiation region can come from the transcription initiation region or the structural gene.

[0024] An application of a TeLCYE protein or a TeLCYE protein-related biomaterial, wherein the application is any one of the following applications:

[0025] (1) Increase the content of α-carotene in plants;

[0026] (2) preparing products that increase the content of α-carotene in plants;

[0027] (3) Increase the lutein content of plants;

[0028] (4) preparing products with increased lutein content in plants;

[0029] (5) Improve plant salt tolerance;

[0030] (6) preparing products that improve plant salt tolerance;

[0031] (7) Improve the rooting of plants under salt stress conditions;

[0032] (8) preparing products for improving the rooting of plants under salt stress conditions;

[0033] (9) Improve the growth of plants under salt stress conditions;

[0034] (10) preparing a product for the growth of plants under salt stress conditions;

[0035] (11), preparing the survival rate of plants under salt stress conditions;

[0036] Wherein, the plant is Arabidopsis thaliana.

[0037] A method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance is to obtain transgenic plants by increasing the expression of a gene encoding a TeLCYE protein in a target plant and / or the content of the TeLCYE protein and / or the activity of the TeLCYE protein.

[0038] As a preferred embodiment of the present invention, the content of α-carotene and / or lutein and the salt tolerance of the transgenic plant are higher than those of the target plant.

[0039] As a preferred embodiment of the present invention, the method for increasing the expression level of the gene encoding the TeLCYE protein in the target plant and / or the content of the TeLCYE protein and / or the activity of the TeLCYE protein is to express or overexpress the TeLCYE protein in the target plant.

[0040] As a preferred embodiment of the present invention, the method for expressing or overexpressing TeLCYE protein is to introduce the gene encoding the TeLCYE protein into the target plant.

[0041] As a preferred embodiment of the present invention, the introduction is carried out by introducing a plant expression vector carrying a gene encoding the TeLCYE protein into the target plant.

[0042] As a preferred embodiment of the present invention, the plant expression vector is transformed into plant cells or tissues by conventional biological methods using Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, Agrobacterium-mediated, and the transformed plant cells or tissues are cultivated into plants.

[0043] As a preferred embodiment of the present invention, the plant expression vector is pCAMBIA1301-TeLCYE.

[0044] As a preferred embodiment of the present invention, the improvement of plant quality is mainly reflected in increasing at least one of the α-carotene and lutein content of the plant.

[0045] Beneficial Effects

[0046] Compared with the prior art, the protein encoded by the TeLCYE gene provided by the present invention can improve the accumulation and stress resistance of plant α-carotene and lutein; overexpression of the TeLCYE gene can improve the accumulation and salt tolerance of plant α-carotene and lutein; the content of α-carotene and lutein in transgenic plants is significantly increased compared with wild-type Arabidopsis plants; the content of α-carotene is 1.13 times that of wild-type plants; the content of lutein is 1.13 times and 1.07 times that of wild-type plants; under salt stress, the transgenic plants show a good growth state, and the root length of the overexpressed transgenic Arabidopsis is 140-150% higher than that of the wild-type Arabidopsis; the TeLCYE protein and its encoding gene provided by the present invention have important application value in the research of improving the content of carotenoids and stress resistance of plants. The present invention will have broad application space and market prospects in the field of agriculture. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 It is the verification diagram of the recombinant plasmid map and the result of restriction digestion of the target gene fragment; among them, lane 1 is the EcoRV restriction digestion band of the target fragment: 1106bp, 2161bp, 2992bp, 5324bp;

[0048] Figure 2 PCR identification of T1 generation TeLCYe transgenic Arabidopsis thaliana; M is DL2000DNAMarker; + is positive control; - is negative control; O is wild-type Arabidopsis thaliana; 1-14 are T1 generation transgenic Arabidopsis thaliana lines;

[0049] Figure 3 This is a comparison of root growth between TeLCYE transgenic and wild-type Arabidopsis under salt stress. DETAILED DESCRIPTION

[0050] The present invention will be further described with reference to the embodiments and the accompanying drawings.

[0051] As Example 1 of the present invention, a TeLCYE protein is provided, wherein the TeLCYE protein is derived from marigold; the TeLCYE protein is a protein having an amino acid sequence as shown in SEQ ID NO: 1. The amino acid sequence shown in SEQ ID NO: 1 is:

[0052] .

[0053] As Example 2 of the present invention, the TeLCYE protein can be artificially synthesized, or its encoding gene can be synthesized first and then obtained by biological expression.

[0054] As Example 3 of the present invention, the TeLCYE protein can be expressed, detected, traced and / or purified by in vitro DNA recombination technology using a protein tag.

[0055] As Example 4 of the present invention, the protein tag includes: a Flag tag, a His tag, a MBP tag, a HA tag, a myc tag, a GST tag and / or a SUMO tag.

[0056] A TeLCYE protein-related biological material, wherein the related biological material is any one of the following related biological materials:

[0057] (1) A nucleic acid molecule encoding a TeLCYE protein, wherein the nucleic acid molecule is a DNA molecule having a nucleotide sequence as shown in SEQ ID NO.2;

[0058] (2) A recombinant vector containing the nucleic acid molecule described in (1);

[0059] (3) A recombinant microorganism containing the nucleic acid molecule described in (1);

[0060] (4) A transgenic plant cell line containing the nucleic acid molecule described in (1);

[0061] (5) Transgenic plant tissue containing the nucleic acid molecule described in (1);

[0062] (6) Transgenic plant organs containing the nucleic acid molecule described in (1);

[0063] (7) A transgenic plant containing the nucleic acid molecule described in (1);

[0064] (8) Tissue culture produced by regenerable cells of transgenic plants containing the nucleic acid molecule described in (1);

[0065] (9) Protoplasts produced by tissue culture containing the nucleic acid molecule described in (1).

[0066]

[0067] As Example 5 of the present invention, a recombinant vector containing the TeLCYE encoding gene expression cassette can be constructed using a plant expression vector. The plant expression vector can be a Gateway system vector or a binary Agrobacterium vector, such as pGWB411, pGWB412, pGWB405, pBin438, pCAMBIA1300, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1301, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb. When using TeLCYE to construct a recombinant vector, any enhanced, constitutive, tissue-specific or inducible promoter can be added before its transcription initiation nucleotide, such as cauliflower mosaic virus (CAMV) 35S promoter, ubiquitin gene Ubiqutin promoter (pUbi), etc., which can be used alone or in combination with other plant promoters; in addition, when using the gene of the present invention to construct a plant expression vector, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signal and the start codon are extensive and can be natural or synthetic. The translation initiation region can come from the transcription initiation region or the structural gene.

[0068] An application of a TeLCYE protein or a TeLCYE protein-related biomaterial, wherein the application is any one of the following applications:

[0069] (1) Increase the content of α-carotene in plants;

[0070] (2) preparing products that increase the content of α-carotene in plants;

[0071] (3) Increase the lutein content of plants;

[0072] (4) preparing products with increased lutein content in plants;

[0073] (5) Improve plant salt tolerance;

[0074] (6) preparing products that improve plant salt tolerance;

[0075] (7) Improve the rooting of plants under salt stress conditions;

[0076] (8) preparing products for improving the rooting of plants under salt stress conditions;

[0077] (9) Improve the growth of plants under salt stress conditions;

[0078] (10) preparing a product for the growth of plants under salt stress conditions;

[0079] (11), preparing the survival rate of plants under salt stress conditions;

[0080] Wherein, the plant is Arabidopsis thaliana.

[0081] A method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance is to obtain transgenic plants by increasing the expression of a gene encoding a TeLCYE protein in a target plant and / or the content of the TeLCYE protein and / or the activity of the TeLCYE protein.

[0082] As Example 6 of the present invention, the content of α-carotene and / or lutein and the salt tolerance of the transgenic plant are higher than those of the target plant.

[0083] As Example 7 of the present invention, the method for increasing the expression level of the gene encoding the TeLCYE protein in the target plant and / or the content of the TeLCYE protein and / or the activity of the TeLCYE protein is to express or overexpress the TeLCYE protein in the target plant.

[0084] As Example 8 of the present invention, the method for expressing or overexpressing TeLCYE protein is to introduce the gene encoding the TeLCYE protein into the target plant.

[0085] As Example 9 of the present invention, the introduction is carried out by introducing a plant expression vector carrying a gene encoding the TeLCYE protein into the target plant.

[0086] As Example 10 of the present invention, the plant expression vector is transformed into plant cells or tissues by conventional biological methods using Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, and Agrobacterium-mediated, and the transformed plant cells or tissues are cultivated into plants.

[0087] As Example 11 of the present invention, the plant expression vector is pCAMBIA1301-TeLCYE.

[0088] As Example 12 of the present invention, the improvement of plant quality is mainly reflected in increasing at least one of the α-carotene and lutein content of the plant.

[0089] As Example 13 of the present invention, the marigold TeLCYE protein and its encoding gene were obtained.

[0090] The nucleotide sequence shown in SEQ ID NO: 2 was synthesized by artificial chemical synthesis (commissioned to Shanghai Bioengineering Co., Ltd., the same below), and the protective base CGC and NdeI restriction site were added to the 5' end of the nucleotide sequence shown in SEQ ID NO: 2, and the protective base CCG and XhoI restriction site were added to the 3' end to obtain the corresponding gene fragment.

[0091] As Example 14 of the present invention, a TeLCYE gene overexpression vector was constructed.

[0092] Primers TeLCYE-F / R were used to amplify the TeLCYE sequence without a signal peptide using the synthetic TeLCYE gene as a template. Primers AtLCYETP-F / R were used to amplify the AtLCYE signal peptide sequence using the Arabidopsis genome as a template. The two amplification products were mixed at a molar ratio of 1:1 and amplified using primers TeLCYE-F / R as a template to obtain the TeLCYE sequence containing the Arabidopsis signal peptide. Subsequently, the vector was amplified using p35s-F / R, and pET-gfp was amplified using gfp-F / R. The fragment was connected to the vector using homologous recombination, and the TeLCYE gene was assembled into the backbone pET-gfp using the ClonExpress Ultra homologous recombination kit (Vazyme Biotech, Nanjing, China). The recombinant strain was obtained after incubation at 50°C for 15 minutes. Plasmid p35s-At-LCYE was obtained after correct sequencing using sequencing primers E1, E2, and E3. Figure 1 shown.

[0093] As Example 15 of the present invention, TeLCYE gene was transformed into Arabidopsis thaliana.

[0094] The plant expression vector p35s-At-LCYE of the marigold TeLCYE gene constructed in Example 14 was transformed into Arabidopsis thaliana by the floral dipping method. The specific method is as follows:

[0095] 1. Agrobacterium Transformation

[0096] Take 1 μL of plasmid and add it to 50 μL of GV3101 Agrobacterium competent cells. Mix thoroughly and then transfer to an electroporation cup. After electroporation, add 1 mL of LB liquid culture medium and mix thoroughly and then transfer to a 1.5 mL centrifuge tube. Incubate in a shaker at 30°C and 180 rpm for 30 min. Pipette 50 μL of the activated Agrobacterium liquid and inoculate it on LB solid culture medium. Incubate in the dark at 30°C for 48 h.

[0097] 2. Agrobacterium infection

[0098] Pick Agrobacterium in the resuspension, prepare Agrobacterium resuspension with OD600 = 0.8-1.2, add silwet-77 to a concentration of 0.02%, dip all the inflorescences of Arabidopsis material in the bacterial solution for 2-3s, seal the film to keep the humidity >90%, and culture in the dark at 25℃ for 24h. The infection cycle is 7 days, and a total of 3 times of infection. Place the infected seedlings in 23℃ 16h / 8h light / dark culture until they set seeds; gently rub the mature fruit pods onto clean white paper, wrap them up and dry them at 37℃ for 24h. After drying, sieve with a 60-mesh sieve and store the clean seeds at 4℃. When the positive seedlings are about 20 days long, use the CTAB method to extract Arabidopsis genomic DNA and perform PCR detection. The detection method is the same as that of Agrobacterium bacteria detection.

[0099] As Example 16 of the present invention, TeLCYE gene transgenic Arabidopsis plants were detected.

[0100] 1. Screening of Arabidopsis seeds

[0101] When the T0 positive seedlings grow to the seed stage, the harvested positive seedling seeds are dried, disinfected with 75% ethanol for 10 min, washed 2-3 times with sterile water, and evenly spread on 1 / 2MS solid plates containing hygromycin (25 μg / mL) resistance, placed at 4°C for 3 days, and then the plates are taken out and placed at 23-25°C, 16h / 8h light / dark culture for 15 days.

[0102] 2. PCR Identification of Transgenic Arabidopsis

[0103] When the Arabidopsis seedlings are about 15 days old, the number of resistant seedlings is counted and transplanted to soil culture medium for further cultivation. When the seedlings grow to 6 true leaves, the genomic DNA of Arabidopsis leaves is extracted by CTAB method, and PCR detection is performed with wild-type Arabidopsis leaves as negative control. The detection method is the same as that of Agrobacterium tumefaciens. Figure 2 shown.

[0104] As Example 17 of the present invention, the carotenoid content in leaves of TeLCYE gene transgenic Arabidopsis plants was determined by high performance liquid chromatography.

[0105] Weigh about 0.1g of leaves, place in liquid nitrogen and grind thoroughly, add 600μL chloroform, vortex and mix, centrifuge for 5min. Collect the centrifugal supernatant and add 400μL saturated KOH methanol solution, vortex and saponify at room temperature for 30min, centrifuge for 5min. Collect the centrifugal supernatant and blow dry with nitrogen at room temperature. Add 50μL ethyl acetate to dissolve, mix and use for HPLC analysis.

[0106] Table 1 Carotenoid content of wild type and TeLCYE transgenic strains

[0107]

[0108] As Example 18 of the present invention, the salt tolerance of TeLCYE gene transgenic Arabidopsis plants was identified.

[0109] Transgenic Arabidopsis and wild-type seeds were sterilized and sown and subcultured on 1 / 2MS medium containing 100mM NaCl. After 1-2 weeks of stress culture, the growth and rooting of Arabidopsis plants were observed. The results showed that under salt stress conditions, Figure 3 The overexpression of Arabidopsis and the wild-type materials became smaller due to the presence of salt stress; however, the overexpression of Arabidopsis had a relatively good growth state compared with the wild-type WT. The growth potential data showed that under salt stress, the root length and fresh weight of the overexpression of Arabidopsis were 1.2-1.3 times and 1.5-1.7 times higher than those of the wild-type WT materials, respectively; indicating that overexpression of the TeLCYE gene significantly improved the salt tolerance of transgenic Arabidopsis plants.

[0110] Table 2 Comparison of leaf growth traits between TeLCYE transgenic Arabidopsis and wild-type Arabidopsis under salt stress (100 mM NaCl)

[0111]

[0112]

[0113] The preferred embodiments of the present invention are described above with reference to the accompanying drawings, but the scope of the rights of the present invention is not limited thereto. Any modification, equivalent substitution and improvement made by a person skilled in the art without departing from the scope and essence of the present invention should be within the scope of the rights of the present invention.

Claims

1. A TeLCYE protein, characterized in that: The TeLCYE protein is derived from marigold; the TeLCYE protein is a protein with an amino acid sequence as shown in SEQ ID NO:

1.

2. A TeLCYE protein according to claim 1, characterized in that: The TeLCYE protein can be expressed, detected, traced and / or purified by in vitro DNA recombinant technology using a protein tag.

3. A TeLCYE protein according to claim 2, characterized in that: The protein tags include: Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag and / or SUMO tag.

4. A TeLCYE protein-related biomaterial, characterized in that: The relevant biological material is any one of the following relevant biological materials: (1) A nucleic acid molecule encoding a TeLCYE protein, wherein the nucleic acid molecule is a DNA molecule having a nucleotide sequence as shown in SEQ ID NO.2; (2) A recombinant vector containing the nucleic acid molecule described in (1); (3) A recombinant microorganism containing the nucleic acid molecule described in (1); (4) A transgenic plant cell line containing the nucleic acid molecule described in (1); (5) Transgenic plant tissue containing the nucleic acid molecule described in (1); (6) Transgenic plant organs containing the nucleic acid molecule described in (1); (7) A transgenic plant containing the nucleic acid molecule described in (1); (8) Tissue culture produced by regenerable cells of transgenic plants containing the nucleic acid molecule described in (1); (9) Protoplasts produced by tissue culture containing the nucleic acid molecule described in (1).

5. An application of TeLCYE protein or a biomaterial related to TeLCYE protein, characterized in that: The application is any one of the following applications: (1) Increase the content of α-carotene in plants; (2) preparing products that increase the content of α-carotene in plants; (3) Increase the lutein content of plants; (4) preparing products with increased lutein content in plants; (5) Improve plant salt tolerance; (6) preparing products that improve plant salt tolerance; (7) Improve the rooting of plants under salt stress conditions; (8) preparing products for improving the rooting of plants under salt stress conditions; (9) Improve the growth of plants under salt stress conditions; (10) preparing a product for the growth of plants under salt stress conditions; (11), preparing the survival rate of plants under salt stress conditions; Wherein, the plant is Arabidopsis thaliana.

6. A method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance, characterized in that: The method is to obtain a transgenic plant by increasing the expression amount of a gene encoding a TeLCYE protein in a target plant and / or the content of the TeLCYE protein and / or the activity of the TeLCYE protein; the content of α-carotene and / or lutein and salt tolerance of the transgenic plant are higher than those of the target plant.

7. A method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance according to claim 6, characterized in that: The method for increasing the expression amount of the gene encoding the TeLCYE protein in the target plant and / or the content of the TeLCYE protein and / or the activity of the TeLCYE protein is to express or overexpress the TeLCYE protein in the target plant.

8. The method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance according to claim 7, characterized in that: The method for expressing or over-expressing TeLCYE protein is to introduce the coding gene of TeLCYE protein into the target plant.

9. The method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance according to claim 8, characterized in that: The introduction is carried out by introducing a plant expression vector carrying a gene encoding the TeLCYE protein into the target plant.

10. A method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance according to claim 9, characterized in that: The plant expression vector is transformed into plant cells or tissues by conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, and Agrobacterium-mediated transformation, and the transformed plant cells or tissues are cultivated into plants.

11. A method for cultivating transgenic plants with increased α-carotene and / or lutein content and salt tolerance according to claim 10, characterized in that: The plant expression vector is pCAMBIA1301-TeLCYE.

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