An artificial combination module and its application in improving the stress resistance and herbicide resistance of plants

Abstract

The present disclosure relates to an artificial combined module and its application in improving the stress resistance and herbicide resistance of plants. The nucleotide sequence of the artificial combined module is shown in SEQ ID NO.1. The artificial combined module is inserted with a stress resistance functional module and a herbicide resistance functional module. The present disclosure combines the stress resistance functional module and the herbicide resistance functional module through biosynthesis technology to assemble a functional module with dual functions of stress resistance and herbicide resistance, which has good application prospects in cultivating plants with stress resistance and herbicide resistance.

Description

Technical Field

[0001] The present disclosure relates to the field of biotechnology, and in particular, to an artificial combination module and its application in improving plant stress resistance and herbicide resistance. Background Art

[0002] Soil salinization, frequent droughts and prolonged high temperatures are the most destructive abiotic stresses in global agriculture, which significantly reduce agricultural productivity through adverse effects on seed germination, plant growth and development, plant vitality and crop yields. According to conservative estimates, droughts in early spring and late autumn in most parts of the north and the south and frequent and persistent high temperatures in summer can cause crop failure in years with severe damage to crop production. However, weeds have strong adaptability, and extreme weather provides them with more growth space and resources. For weeds with better tolerance to extreme weather, they will further occupy crop space.

[0003] At present, the impact of extreme weather on agriculture is mainly addressed through physical or chemical methods to increase crop yields, such as applying organic fertilizers, improving soil structure, spraying plant stimulants to increase plant tolerance, and using herbicides to control weed growth. However, the application of herbicides can also affect the growth of crops. The extensive use of herbicides has increased the cost of agricultural production, and has also failed to fundamentally solve the existing problems.

[0004] Using synthetic biology to construct artificial combination modules to improve plant stress resistance is a widely used strategy in agriculture. At present, artificial combination modules are only targeted at drought resistance, salt resistance and high temperature resistance. In order to further promote the development of agriculture, it is urgent to provide an artificial combination module that is resistant to herbicides, drought, salt and high temperature. Summary of the invention

[0005] The purpose of the present disclosure is to improve the stress resistance and herbicide resistance of plants.

[0006] In order to achieve the above-mentioned object, the first aspect of the present disclosure provides an artificial combination module, the nucleotide sequence of the artificial combination module is shown as SEQ ID NO.1.

[0007] Optionally, the artificial combination module is inserted with a stress resistance function module and a herbicide resistance function module;

[0008] The nucleotide sequence of the stress resistance functional module is from position 1 to position 6674 of SEQ ID NO.1;

[0009] The nucleotide sequence of the herbicide resistance functional module is from position 6710 to position 11673 of SEQ ID NO.1.

[0010] Optionally, the stress resistance functional module includes a first promoter, an RBS binding site, a first spacer sequence, a SipA protein encoding gene, a second spacer sequence, a DosH protein encoding gene and a first terminator connected sequentially from upstream to downstream;

[0011] Wherein, the first promoter is the Gmubi promoter; and the RBS binding site is rpiL.

[0012] Optionally, the herbicide resistance functional module includes a second promoter, a third spacer sequence, a GR79 gene, a fourth spacer sequence, a GAT gene, a fifth spacer sequence, a second terminator, a sixth spacer sequence, a third promoter, a seventh spacer sequence, a DICX4 gene, an eighth spacer sequence and a third terminator connected sequentially from upstream to downstream;

[0013] Among them, the second promoter is 2xpCaMV35S promoter; the third promoter is p-FMV promoter.

[0014] The second aspect of the present disclosure provides application of the artificial combination module described in the first aspect in improving the stress resistance and herbicide resistance of plants.

[0015] Optionally, the improving the stress resistance and herbicide resistance of the plants is to improve the stress resistance and herbicide resistance of the plants during plant variety breeding.

[0016] Optionally, the improving the stress resistance of the plant is to improve the drought resistance, high salt resistance and high temperature resistance of the plant cells.

[0017] The third aspect of the present disclosure provides a recombinant expression vector, wherein the recombinant expression vector is a backbone vector into which the artificial combination module described in the first aspect is inserted.

[0018] A fourth aspect of the present disclosure provides a transformant, into which the artificial combination module described in the first aspect or the recombinant expression vector described in the third aspect is inserted.

[0019] A fifth aspect of the present disclosure provides a method for improving the stress resistance and herbicide resistance of a plant, the method comprising:

[0020] Introducing the recombinant expression vector described in the third aspect into a target plant and / or a target plant seed;

[0021] Alternatively, the transformant described in the fourth aspect is used to infect the target plant and / or target plant seeds.

[0022] Through the above technical solution, the artificial combination module disclosed in the present invention includes a stress resistance function module and a herbicide resistance function module; the present invention uses biosynthesis technology to combine the stress resistance function module and the herbicide resistance function module to assemble a functional module with dual functions of stress resistance and herbicide resistance. It has a good application prospect in cultivating plants with stress resistance and herbicide resistance.

[0023] Other features and advantages of the present disclosure will be described in detail in the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The accompanying drawings are used to provide a further understanding of the present disclosure and constitute a part of the specification. Together with the following specific embodiments, they are used to explain the present disclosure but do not constitute a limitation of the present disclosure. In the accompanying drawings:

[0025] Figure 1 It is a vector construction diagram of the artificial combination module disclosed in the present invention.

[0026] Figure 2 It is a construction diagram of the artificial combination module of the present disclosure.

[0027] Figure 3 This is a picture of the transgenic rapeseed plant that is tolerant to high salt and drought.

[0028] Figure 4 This is a picture of the herbicide-resistant transgenic rapeseed plant disclosed in the present invention.

[0029] Figure 5 This is a picture of the transgenic corn plants disclosed herein that are resistant to high salt and drought. DETAILED DESCRIPTION

[0030] The specific implementation of the present disclosure is described in detail below in conjunction with the accompanying drawings. It should be understood that the specific implementation described herein is only used to illustrate and explain the present disclosure, and is not used to limit the present disclosure.

[0031] A first aspect of the present disclosure provides an artificial combination module, the nucleotide sequence of the artificial combination module is shown as SEQ ID NO.1.

[0032] The artificial combination module of the present disclosure is inserted with a stress resistance function module and a herbicide resistance function module; wherein the nucleotide sequence of the stress resistance function module is from position 1 to position 6674 of SEQ ID NO.1; the nucleotide sequence of the herbicide resistance function module is from position 6710 to position 11673 of SEQ ID NO.1. The stress resistance function module and the herbicide resistance function module are connected by the sequence shown in positions 6675 to 6709 of SEQ ID NO.1.

[0033] The present invention utilizes modern synthetic biology design methods to optimize and transform stress-resistant elements. Through the artificial design of protein functional elements, the tissue specificity of promoters and the stress response design, an anti-herbicide module that specifically responds to different types of herbicides and an anti-stress functional module that responds to different abiotic stresses are artificially constructed, and the above two modules are assembled to form an artificial combination module with dual effects of stress resistance and herbicide resistance for intelligent response and directional expression, named EgDICW. The full length of the artificial combination module disclosed in the present invention is 11673bp. It has good application prospects in cultivating plants with stress resistance and herbicide resistance. Among them, stress resistance refers to the ability to resist drought, high salt and high temperature.

[0034] In one embodiment of the present disclosure, the stress resistance functional module includes a first promoter, an RBS binding site, a first spacer sequence, a SipA protein encoding gene, a second spacer sequence, a DosH protein encoding gene and a first terminator connected sequentially from upstream to downstream;

[0035] Wherein, the first promoter is the Gmubi promoter; and the RBS binding site is rpiL.

[0036] According to the present disclosure, the nucleotide sequence of the Gmubi promoter is positions 1 to 1416 of SEQ ID NO.1, the nucleotide sequence of the RBS binding site rpiL is positions 1417 to 1785 of SEQ ID NO.1, the nucleotide sequence of the first spacer sequence is positions 1786 to 1851 of SEQ ID NO.1, the nucleotide sequence of the SipA protein encoding gene is positions 1852 to 5355 of SEQ ID NO.1, the nucleotide sequence of the second spacer sequence is positions 5356 to 5421 of SEQ ID NO.1, the nucleotide sequence of the DosH protein encoding gene is positions 5422 to 6318 of SEQ ID NO.1, and the nucleotide sequence of the first terminator is positions 6319 to 6674 of SEQ ID NO.1.

[0037] In one embodiment of the present disclosure, the herbicide resistance functional module includes a second promoter, a third spacer sequence, a GR79 gene, a fourth spacer sequence, a GAT gene, a fifth spacer sequence, a second terminator, a sixth spacer sequence, a third promoter, a seventh spacer sequence, a DICX4 gene, an eighth spacer sequence and a third terminator connected sequentially from upstream to downstream;

[0038] Among them, the second promoter is 2xpCaMV35S promoter; the third promoter is p-FMV promoter.

[0039] According to the present disclosure, the nucleotide sequence of the 2xpCaMV35S promoter is 6710th to 7489th of SEQ ID NO.1, the nucleotide sequence of the third spacer sequence is 7490th to 7496th of SEQ ID NO.1, the nucleotide sequence of the GR79 gene is 7497th to 9065th of SEQ ID NO.1, the nucleotide sequence of the fourth spacer sequence is 9066th to 9128th of SEQ ID NO.1, the nucleotide sequence of the GAT gene is 9129th to 9569th of SEQ ID NO.1, the nucleotide sequence of the fifth spacer sequence is 9570th to 9575th of SEQ ID NO.1, the nucleotide sequence of the second terminator is 9576th to 9750th of SEQ ID NO.1, the nucleotide sequence of the sixth spacer sequence is 9751st to 9758th of SEQ ID NO.1, and the nucleotide sequence of the p-FMV promoter is SEQ ID NO.1 is from 9759 to 10322, the nucleotide sequence of the seventh spacer sequence is from 10323 to 10326 of SEQ ID NO.1, the nucleotide sequence of the DICX4 gene is from 10327 to 11412 of SEQ ID NO.1, the nucleotide sequence of the eighth spacer sequence is from 11413 to 11420 of SEQ ID NO.1, and the nucleotide sequence of the third terminator is from 11421 to 11673 of SEQ ID NO.1.

[0040] The second aspect of the present disclosure provides application of the artificial combination module described in the first aspect in improving the stress resistance and herbicide resistance of plants.

[0041] In one embodiment of the present disclosure, the improving the stress resistance and herbicide resistance of plants is to improve the stress resistance and herbicide resistance of plants during plant variety breeding. The plants can be selected from corn, rapeseed, etc., preferably corn and / or rapeseed.

[0042] In one embodiment of the present disclosure, the improving the stress resistance of plants is to improve the drought resistance, high salt resistance and high temperature resistance of plant cells.

[0043] In the present disclosure, the herbicide in the herbicide resistance includes glyphosate and / or dicamba. High salt tolerance refers to a salt concentration of 300 mM; high temperature tolerance refers to a temperature of 30°C-45°C.

[0044] The third aspect of the present disclosure provides a recombinant expression vector, wherein the recombinant expression vector is a backbone vector into which the artificial combination module described in the first aspect is inserted.

[0045] According to the present disclosure, the backbone vector may be pJET, pBI-121 or PLUS.

[0046] In the present disclosure, by constructing a recombinant expression vector, expression frames of stress-resistant genes (i.e., SipA protein encoding gene and DosH protein encoding gene) and expression frames of herbicide-resistant genes (i.e., GR79 gene, GAT gene and DICX4 gene) are formed, so that the stress-resistant genes and herbicide-resistant genes are expressed in the target plants, thereby making the target plants have stress-resistant and herbicide-resistant functions.

[0047] A fourth aspect of the present disclosure provides a transformant, into which the artificial combination module described in the first aspect or the recombinant expression vector described in the third aspect is inserted.

[0048] According to the present disclosure, the host of the transformant is Agrobacterium tumefaciens EHA105.

[0049] A fifth aspect of the present disclosure provides a method for improving the stress resistance and herbicide resistance of a plant, the method comprising:

[0050] Introducing the recombinant expression vector described in the third aspect into a target plant and / or a target plant seed;

[0051] Alternatively, the transformant described in the fourth aspect is used to infect the target plant and / or target plant seeds.

[0052] In the present disclosure, the infected target plant may be a callus of the infected target plant.

[0053] The present disclosure is further illustrated below by way of examples, but the present disclosure is not limited thereby.

[0054] Unless otherwise specified, the raw materials, reagents, instruments and equipment involved in the embodiments of the present disclosure can be obtained by purchase.

[0055] If no specific experimental conditions are specified in the examples disclosed herein, all are based on conventional conditions familiar to those skilled in the art, such as the conditions described in Sambrook et al. Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer.

[0056] Cloning vector pJET: a commercial product from ThermoFisher;

[0057] Shuttle vector: pBI-121: stored in this laboratory;

[0058] Agrobacterium tumefaciens EHA105: maintained in this laboratory.

[0059] Example 1

[0060] This example is used to illustrate the preparation of recombinant Agrobacterium tumefaciens of artificial combination modules:

[0061] 1. Experimental methods:

[0062] like Figure 1 and 2 As shown in the figure, different types of herbicide resistance functional modules were designed through synthetic biology, and functional modules that specifically respond to different abiotic stresses were designed and constructed, and a new stress resistance functional circuit with intelligent response and directional expression was assembled, named EgDICW (also known as artificial combination module). The full-length nucleic acid sequence of the stress resistance functional circuit EgDICW was obtained by artificial chemical synthesis. Its size is 11673bp, which was cloned into the vector pJET to construct a recombinant cloning plasmid pJET-EGDICW containing a complete stress resistance functional circuit, and sequenced and verified.

[0063] Then pass Eco RI and Hin The stress-resistant circuit EGICW fragment containing sticky ends and the shuttle vector pBI-121 vector fragment were obtained by double digestion with dIII. The stress-resistant circuit EGICW was connected to the pBI-121 vector to construct a plant expression vector pBI-EGDICW. The expression vector was transformed into Agrobacterium tumefaciens EHA105, and positive recombinant strains were screened by kanamycin antibiotic resistance, and verified by colony PCR sequencing.

[0064] 2. Experimental results:

[0065] The full-length nucleic acid sequence of the stress-resistant functional circuit EgDICW was obtained by artificial chemical synthesis, with a total length of 11673bp.

[0066] The plant expression vector pBI-EGDICW containing the functional line EGDICW was successfully constructed and transformed into Agrobacterium tumefaciens EHA105.

[0067] The insertion sequence was verified to be correct by PCR, restriction digestion and sequencing, and the strain was named EHA-EGDICW.

[0068] 3. Experimental conclusion:

[0069] The construction of recombinant Agrobacterium tumefaciens EHA-EGDICW expressing the stress resistance functional circuit EGDICW was completed.

[0070] Example 2

[0071] This example is used to illustrate the construction of transgenic rapeseed.

[0072] The target plant selected in this example is Brassica napus, and rapeseed seeds 84100-18 are stored in this laboratory.

[0073] 1. Experimental methods:

[0074] Take rapeseed seeds and use 75% ethanol and 0.1% HgCl 2 Soak and sterilize, place evenly in plant tissue culture medium, and culture in a 24ºC tissue culture room for one week. Use sterilized surgical scissors to remove the hypocotyls of rapeseed seedlings, place them on the pre-culture medium, and culture them under light for 2-3 days to pre-culture explants.

[0075] Transfer and activate the recombinant Agrobacterium strain EHA-EGDICW expressing the stress resistance function line, collect the strain by centrifugation and resuspend it to OD 600 =1.0. Soak the pre-cultured explants in Agrobacterium solution for 90 seconds, transfer them to the co-culture medium after drying, and culture them in the dark for 2-3 days. Then transfer the well-growing explants to the induction medium for culture.

[0076] Select explants with good callus growth and transfer them to MS medium screening medium supplemented with kana antibiotics, culture them under light for 45-50 days, and then differentiate them into buds. Transfer the differentiated budding callus to MS rooting medium and culture them under light for 2 weeks. When the roots appear and the stems grow 4-5 cm, transfer them to culture soil for seedling training. After acclimatization, transplant them to the greenhouse, and PCR test positive rapeseed seedlings.

[0077] 2. Experimental results:

[0078] The stress-resistant functional circuit EGICW was transformed into rapeseed using the Agrobacterium-mediated explant co-cultivation method. After the infected rapeseed explants went through induction culture, screening culture, rooting culture, seedling training and transplantation, and PCR verification, we finally obtained the transgenic rapeseed Bn-EGDICW expressing the stress-resistant functional circuit, which can be used for subsequent stress resistance research.

[0079] 3. Experimental conclusion:

[0080] Through the Agrobacterium-mediated transformation method, the stress-resistant functional line EgDICW rapeseed Bn-EGDICW was finally obtained.

[0081] Example 3

[0082] This example is used to illustrate the stress resistance and herbicide resistance of rapeseed Bn-EGDICW in Example 2.

[0083] The experimental group used the transgenic rapeseed obtained in Example 2 (ie, transgenic rapeseed seeds); the control group used non-transgenic rapeseed (ie, wild-type seeds).

[0084] 1. Experimental methods:

[0085] Salt stress was simulated by adding NaCl; drought stress was simulated by adding polyethylene glycol PEG-6000; and herbicide stress was simulated by a mixture of glyphosate and dicamba.

[0086] The transgenic rapeseed seeds and wild-type seeds that have been identified as positive are cultured in MS solid culture. After the seedlings grow true leaves, they are transplanted into plastic pots filled with substrates and watered with MS nutrient solution. When the seedlings grow 5-6 true leaves, they are subjected to stress treatment.

[0087] Among them, the high salt stress treatment includes: watering the plants with 20 mL of salt solution with a concentration of 300 mM every day.

[0088] Drought stress treatment consisted of watering the plants with 20 mL of polyethylene glycol at a concentration of 15% per day.

[0089] Herbicide stress included: spraying a mixed herbicide of glyphosate and dicamba purchased from the market (purchased from Weifang Zhongnong United Chemical Co., Ltd.) when the rapeseed plants were in the four-leaf stage, and the concentration used was according to the instructions.

[0090] 2. Experimental results:

[0091] The results of coercion are as follows Figure 3 and 4 As shown; the research results showed that under normal growth conditions before treatment, there was no difference in the growth status of transgenic rapeseed Bn-EGDICW and wild-type rapeseed, and the agronomic traits were not affected.

[0092] exist Figure 3 In the figure, the left side shows the growth status of non-transgenic rapeseed on the 14th day in saline-alkali soil, and the right side shows the growth status of transgenic rapeseed on the 14th day in saline-alkali soil. After 7 days of 15% severe drought stress, the wild-type rapeseed began to wither and wilt, while the transgenic rapeseed Bn-EGDICW slowed down its growth rate, and the growth of leaves and stems was not significantly affected; after 14 days of drought treatment, the wild-type rapeseed had completely withered, and the transgenic rapeseed began to wilt to a certain extent. In the high-salt stress experiment, after 7 days of 300mM NaCl stress treatment, the wild-type rapeseed suffered from severe water loss and drying, while some leaves of the transgenic rapeseed Bn-EGDICW turned yellow, and its growth condition was significantly better than that of the wild type; after 14 days of high-salt treatment, the wild-type rapeseed had basically dried up and died, while the transgenic rapeseed Bn-EGDICW had curled leaves, wilted stems, and slowed growth.

[0093] exist Figure 4 In the figure, the left side shows the growth status of non-transgenic rapeseed on the 14th day after spraying with herbicides, and the right side shows the growth status of transgenic rapeseed on the 14th day after spraying with herbicides. The wild-type rapeseed was completely killed by the mixed herbicide treatment of glyphosate and dicamba, while the transgenic rapeseed Bn-EGDICW was able to resist two different herbicides.

[0094] 3. Experimental conclusion:

[0095] The stress resistance genes in the artificial combination module disclosed in the present invention were successfully expressed in rapeseed, which significantly improved the high salt tolerance, drought resistance, and herbicide resistance of the rapeseed plants.

[0096] Example 4

[0097] This example is used to illustrate the construction of transgenic corn.

[0098] 1. Experimental methods:

[0099] Corn seeds were peeled and lysed with 75% ethanol and 0.1% HgCl 2 Soak and sterilize, place evenly on plant tissue culture medium, and culture in a 24ºC tissue culture room for 2 weeks. Use sterilized surgical scissors to remove rice callus, place on pre-culture medium, and culture in the dark for 2 weeks.

[0100] Transfer and activate the recombinant Agrobacterium strain EHA-EGDICW expressing the stress resistance circuit, collect the strain by centrifugation and resuspend it to OD600=1.0. Soak the pre-cultured explants in the Agrobacterium bacterial solution for 30 minutes, transfer them to the co-culture medium after drying, and culture them in the dark for 2-3 days. Then transfer them to the induction medium for culture.

[0101] Select callus tissue and transfer it to MS screening medium supplemented with kana antibiotics, culture it in the dark for 2 weeks, screen it again and culture it in the dark for 2 weeks, and then culture it for 1 week. Transfer the differentiated callus to MS rooting medium, wait for the root system to appear and the stem to grow 4-5cm, and then transfer it to the greenhouse, and test the positive corn seedlings by PCR.

[0102] 2. Experimental results:

[0103] The stress-resistant functional circuit EgDICW was transformed into corn through the Agrobacterium-mediated callus co-culture method. After the infection of corn callus tissue, induction culture, resistance screening culture, rooting culture and seedling transplantation, and PCR verification, the transgenic corn Co-EGDICW expressing the stress-resistant functional circuit was finally obtained, which can be used for subsequent stress resistance research.

[0104] 3. Experimental conclusion:

[0105] Through the Agrobacterium-mediated transformation method, the stress-resistant functional line EGDICW corn Co-EGDICW was finally obtained.

[0106] Example 5

[0107] This example is used to illustrate the stress resistance and herbicide resistance of corn Co-EGDICW in Example 4.

[0108] The experimental group used the transgenic corn (i.e., transgenic rapeseed seeds) obtained in Example 4; the control group used non-transgenic corn (i.e., wild-type seeds).

[0109] 1. Experimental methods:

[0110] Salt stress was simulated by adding NaCl; drought stress was simulated by adding polyethylene glycol PEG-6000; and herbicide stress was simulated by a mixture of glyphosate and dicamba.

[0111] The stress treatment was carried out by watering. The wild-type corn and positive transgenic corn Co-EGDICW seeds were germinated and cultured in MS medium for seedling emergence. When the corn seedlings grew to 2 leaves and 1 heart, the stress treatment was carried out about 12 days later, and the growth status of the plants was observed.

[0112] Among them, the high salt stress treatment includes: watering the plants with 20 mL of salt solution with a concentration of 300 mM every day.

[0113] Drought stress treatment consisted of watering the plants with 20 mL of polyethylene glycol at a concentration of 15% per day.

[0114] 2. Experimental results:

[0115] The results of coercion are as follows Figure 5 As shown; the study showed that under non-stress conditions, the emergence and growth of transgenic corn Co-EGDICW were no different from those of wild-type rice.

[0116] exist Figure 5 In the figure, the right side shows the growth status of non-GMO corn in saline-alkali soil on the 7th day, and the left side shows the growth status of GMO corn in saline-alkali soil on the 7th day.

[0117] After 7 days of severe drought stress of 15%, the wild-type corn began to wither, yellow, fall leaves and wilt, while the transgenic corn Co-EGDICW slowed down its growth rate, and the growth of leaves and stems was not significantly affected. In the high-salt stress experiment, after 7 days of 300mM NaCl stress treatment, the wild-type corn suffered from severe water loss and withering, while the transgenic corn Co-EGDICW had some yellowing leaves and its growth condition was significantly better than that of the wild type.

[0118] 3. Experimental conclusion:

[0119] The stress-resistant genes in the artificial combination module disclosed in the present invention were successfully expressed in corn, which significantly improved the high-salt and drought resistance of corn plants.

[0120] The preferred embodiments of the present disclosure are described in detail above in conjunction with the accompanying drawings; however, the present disclosure is not limited to the specific details in the above embodiments. Within the technical concept of the present disclosure, a variety of simple modifications can be made to the technical solution of the present disclosure, and these simple modifications all fall within the protection scope of the present disclosure.

[0121] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present disclosure will not further describe various possible combinations.

[0122] In addition, various embodiments of the present disclosure may be arbitrarily combined, and as long as they do not violate the concept of the present disclosure, they should also be regarded as the contents disclosed by the present disclosure.

Claims

1. An artificial combination module, characterized in that: The nucleotide sequence of the artificial combination module is shown in SEQ ID NO.

1.

2. The artificial combination module according to claim 1, wherein: The artificial combination module is inserted with a stress resistance function module and a herbicide resistance function module; The nucleotide sequence of the stress resistance functional module is from position 1 to position 6674 of SEQ ID NO.1; The nucleotide sequence of the herbicide resistance functional module is from position 6710 to position 11673 of SEQ ID NO.

1.

3. The artificial combination module according to claim 2, wherein: The stress resistance functional module includes a first promoter, an RBS binding site, a first spacer sequence, a SipA protein encoding gene, a second spacer sequence, a DosH protein encoding gene and a first terminator connected in sequence from upstream to downstream; Wherein, the first promoter is the Gmubi promoter; and the RBS binding site is rpiL.

4. The artificial combination module according to claim 2, wherein: The herbicide resistance functional module includes a second promoter, a third spacer sequence, a GR79 gene, a fourth spacer sequence, a GAT gene, a fifth spacer sequence, a second terminator, a sixth spacer sequence, a third promoter, a seventh spacer sequence, a DICX4 gene, an eighth spacer sequence and a third terminator connected sequentially from upstream to downstream; Among them, the second promoter is 2xpCaMV35S promoter; the third promoter is p-FMV promoter.

5. Use of the artificial combination module according to any one of claims 1 to 4 in improving the stress resistance and herbicide resistance of plants, characterized in that: The plant is corn and / or rapeseed; The improving the stress resistance of plants is to improve the drought resistance, high salt resistance and high temperature resistance of plant cells.

6. The use according to claim 5, wherein: The method of improving the stress resistance and herbicide resistance of plants is to improve the stress resistance and herbicide resistance of plants during plant variety breeding.

7. A recombinant expression vector, characterized in that: The recombinant expression vector is a backbone vector into which the artificial combination module according to any one of claims 1 to 4 is inserted.

8. A transformant, characterized in that The transformant is inserted with the artificial combination module according to any one of claims 1 to 4 or the recombinant expression vector according to claim 7.

9. A method for improving the stress resistance and herbicide resistance of plants, characterized in that: The method includes: Introducing the recombinant expression vector according to claim 7 into a target plant and / or a target plant seed; Alternatively, using the transformant according to claim 8 to infect a target plant and / or a target plant seed; The plant is corn and / or rapeseed; The improving of the stress resistance of plants is to improve the drought resistance, high salt resistance and high temperature resistance of plant cells.

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