CN116656698A - Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops - Google Patents
Application of corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops Download PDFInfo
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Abstract
The invention belongs to the technical field of plant biology, and discloses application of a corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops, in particular to application in improving drought resistance of corn, and application in germplasm resource improvement, genetic breeding and preparation of corresponding drought resistant transgenic crops. Also disclosed is the expression of the gene in yeast to promote the tolerance of yeast cells to osmotic stress caused by sorbitol, and the overexpression in Arabidopsis to promote germination of transgenic plants under osmotic stress. The invention verifies that the corn gene Zm00001d018037 has the function of improving the drought stress resistance of corn by creating a gene editing mutant, transgenic over-expressing corn genetic material and drought resistance phenotype detection, thereby providing a new technical means for cultivating new strain of drought-resistant corn and creating germplasm resources, and having important agricultural application value and wide practical prospect.
Description
Technical Field
The invention relates to the technical field of biology, in particular to application of a corn gene Zm00001d018037 in improving drought resistance of monocotyledonous crops, and especially in improving drought resistance of corn.
Background
Drought stress seriously affects the growth and development of plants and the yield of crops, and has become an important research topic for stress of plants. Corn is the first crop worldwide, its yield and quality impact human society survival and development, but corn growth and yield are also severely threatened by drought stress. In long-term drought research, components playing important roles in drought stress response of a plurality of plants have been identified, but the research on identification and action pathways of the components is mainly realized by mode plants such as arabidopsis thaliana, and the like, so that the research on important functions and improvement components of drought stress in important crops such as corn, rice and wheat is relatively less, and the identification of the important components and the drought response pathways in response to the drought stress has more theoretical and practical significance.
The protein coded by the corn Zm00001d018037 gene belongs to a NIP channel protein family member, the CDS length of the corn Zm00001d018037 gene is 885 bp, the protein codes for the NIP channel protein with 295 amino acids, and at present, no research report on the gene in the drought stress reaction of corn is seen in the prior art.
Disclosure of Invention
The technical problem to be solved by the invention is how to alleviate the influence of drought stress on monocotyledonous crops, especially on the growth and development of corn and the yield, namely how to improve the drought stress resistance of corn.
The technical scheme of the invention is as follows: the application of the corn gene Zm00001d018037 in improving the drought resistance of monocotyledonous crops is provided, and the application is preferably in improving the drought resistance of corn.
The invention also provides application of the maize gene Zm00001d018037 in germplasm resource improvement, genetic breeding and preparation of corresponding drought-resistant transgenic crops.
Furthermore, the invention also provides a method for improving the drought stress response of monocotyledonous crops, which integrates the corn gene Zm00001d018037 into cells, tissues or organs of the monocotyledonous crops and enables the monocotyledonous crops to be overexpressed, wherein the monocotyledonous crops are preferably corn.
The invention also provides a method for cultivating the high drought-resistant corn, which comprises the following steps: transforming corn gene Zm00001d018037 into corn young embryo, co-culturing, resistance screening, callus induction, subculture and differentiation to obtain T0 generation transgenic corn, and continuous selfing and target gene PCR detection to obtain stable T 3 And (3) obtaining the maize with the excessive expression of Zm00001d018037, namely the maize with high drought resistance, by using the generation homozygous transgenic line.
Since osmotic stress is one of the most important aspects of plants subjected to drought stress, the invention also provides application of the maize gene Zm00001d018037 in improving the osmotic stress tolerance of yeast cells, so that the maize gene Zm00001d018037 is expressed in the yeast, and the osmotic stress tolerance of the yeast cells is improved.
The invention also provides application of the maize gene Zm00001d018037 in improving seed germination capacity under the osmotic stress of arabidopsis, so that the maize gene Zm00001d018037 is expressed in arabidopsis, and the seed germination capacity under the osmotic stress of transgenic arabidopsis is improved.
The prior art predicts that maize gene Zm00001d018037 encodes an NIP channel protein, but no report has been made regarding its role in plant abiotic stress. The inventor discovers that all the gene editing mutant lines cause triplet codon shift by creating a corn gene Zm00001d018037 gene editing mutant line, and reveals the gene editing mutant line of a corn gene Zm00001d018037 through experiments, which shows a drought stress sensitive phenotype, and the corn gene Zm00001d018037 is overexpressed in transgenic corn to promote the enhancement of drought tolerance of the transgenic corn. The gene plays an important role in drought stress response of plants, so that a new technical means is provided for cultivating excellent crop strains with enhanced drought tolerance by using the gene.
The invention has the following beneficial effects: the invention provides the application of the corn gene Zm00001d018037 in improving the drought resistance of monocotyledonous crops, especially in improving the drought resistance of corn, and in germplasm resource improvement, genetic breeding and preparation of corresponding drought-resistant transgenic crops, and also provides the technical scheme that the expression of the gene in yeast promotes the osmotic stress tolerance of yeast cells to sorbitol, and the overexpression in Arabidopsis promotes the germination of transgenic plants under osmotic stress, and provides a new technical means for cultivating new corn varieties with high drought resistance and innovation of excellent corn germplasm resources, so that the invention has important agricultural application value and wide practical prospect.
Drawings
FIG. 1 is an illustration of sequencing and identification of the basic structure, the gene editing target position and the gene editing mutant Zm00001d018037-d1 strain and the Zm00001d018037-d2 strain of the maize gene Zm00001d 018037; wherein A is the description of the basic structure and the gene editing target position of the corn gene Zm00001d018037, B is the description of the sequencing identification of the gene editing type of the corn gene Zm00001d018037-d1 strain, and C is the description of the sequencing identification of the gene editing type of the corn gene Zm00001d018037-d2 strain.
FIG. 2 is an illustration of the detection of drought tolerance, detection of moisture content of plant tissue before and after severe drought treatment, and detection of accumulation of membranous oxidation products (MDA) of plant tissue before and after severe drought treatment for maize gene Zm00001d018037 gene editing mutants (ZmNIP 2a-d1 and ZmNIP2a-d 2) after severe drought treatment; wherein A is the description of drought tolerance detection of maize gene Zm00001d018037 gene editing mutants (ZmNIP 2a-d1 and ZmNIP2a-d 2) after severe drought treatment, B is the description of water content detection of plant tissues before and after severe drought treatment, and C is the description of accumulation detection of membranous oxidation products (MDA) of plant tissues before and after severe drought treatment.
FIG. 3 is a schematic representation of maize genesZm00001d018037Description of PCR detection of transcript levels in overexpressed maize lines.
FIG. 4 is a schematic representation of maize genesZm00001d018037And (5) gene editing and identification of transgenic super-expression genetic material seedling drought resistance phenotype.
FIG. 5 is a diagram showing that expression of ZmNIP2a in yeast promotes osmotic stress tolerance of yeast.
FIG. 6 is an illustration of germination rate statistics after 4 days of osmotic stress tolerance during germination of ZmNIP2a gene-promoted mode plant Arabidopsis thaliana, and overexpression of ZmNIP2a gene; wherein A is an illustration of osmotic stress tolerance during germination of Arabidopsis thaliana which is a ZmNIP2a gene promotion mode, and B is an illustration of germination rate statistics after 4 days of osmotic stress of Arabidopsis thaliana over-expressed by the ZmNIP2a gene.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The methods used in the examples are conventional methods known to those skilled in the art, and the reagents used, if not specified, are commercially available products. The corn gene editing mutant and the transgenic super-expression material are derived from a corn molecular breeding and gene editing platform in a national key laboratory of the stress adaptation and improvement of co-constructed crops of Henan university, and corn B73-329 is taken as an experimental base material.
For the convenience of expression and simplicity and clarity of experimental records, the corn Zm00001d018037 gene is simply expressed as ZmNIP2a, and the nucleotide sequence is shown as NO.1 in the ST.26 sequence table; the single nucleotide addition (+ bp) gene editing mutant of the maize Zm00001d018037 gene is expressed asZmNIP2a-d1，The nucleotide sequence is shown as NO.2 in a ST.26 sequence table; the deletion (-7 bp) gene editing mutant of the maize Zm00001d018037 gene is expressed asZmNIP2a-d2，The nucleotide sequence is shown as NO.3 in the ST.26 sequence table.
Creating gene editing mutant and transgenic super-expression corn genetic material
(1) According to the gene coding sequence of the corn Zm00001d018037 gene, selecting a target (double target, target 1 sequence: ACGCCGTCGGACACATCTC, target 2 sequence: GAAGATGTCCGCCAGCGAC, wherein target 1 is the main component), designing primers according to the target sequence (four primer sequences are Zm00001d018037MT1T2-F: AATAATGGTCTCAGGCGACGCCGTCGGACACATCTC, zm00001d018037MT1T2-F0: GACGCCGTCGGACACATCTCGTTTTAGAGCTAGAAATAGC, zm00001d018037MT1T2-R0: GTCGCTGGCGGACATCTTCCGCTTCTTGGTGCC, zm00001d018037MT1T2-R: ATTATTGGTCTCTAAACGTCGCTGGCGGACATCTTC) respectively, carrying out four-primer PCR amplification by taking an intermediate vector pCBC-MT1T2 as a template, and purifying a PCR amplified fragment. The amplified fragment was subjected to cleavage ligation with the gene editing final vector pBUE411 in the following manner:
taking 5 mu L of the liquid, enzyme cutting, connecting products to convert competent cells of escherichia coli, culturing overnight on an antibiotic LB culture medium containing kanamycin, and screening positive clones. The positive transformants were cultured overnight (37 ℃ C., 220 rpm) on liquid LB medium. And extracting the successfully connected recombinant plasmid DNA, and sequencing to confirm the existence of the designed target sequence in the recombinant vector. And (3) transferring the successfully constructed gene editing vector into an agrobacterium EHA105 strain, submitting the constructed crop adversity adaptation of the Henan university province and improving a national key laboratory corn molecular breeding and gene editing platform to perform genetic material creation work. And breeding the created gene editing T0 generation material to obtain T1 generation gene editing germplasm. And (5) planting a gene editing strain, taking corn leaves of the strain, and extracting DNA.
The SLS extraction method of the strain corn DNA is as follows: taking 0.1 g of the maize leaves of the strain, placing the maize leaves in a 2 mL centrifuge tube which is sterilized in advance and is provided with small steel balls, and quick-freezing the maize leaves in liquid nitrogen; the quick frozen sample was shaken in a mill at 60 Hz frequency for 1 min, 750 μl of SLS DNA extract (1% SLS, 0.02M EDTA (ph=8.0), 0.1M Tris-HCl (ph=8.0), 0.1M NaCl) was added to the sample, and vigorously shaken to mix it thoroughly. Adding 750 mu L of phenol: chloroform: isoamyl alcohol (25:24:1) and fully shaking, standing for 5 min at room temperature, and centrifuging at high speed for 10 min at 4 ℃ and 12,000 rpm; transferring the supernatant to a centrifuge tube of 1.5 mL, adding equal volume of isopropanol, mixing upside down, and standing in a refrigerator at 4deg.C for 30 min; high-speed centrifugation at 12,000 rpm at 4℃for 10 min to precipitate DNA; washing the DNA with 75% ethanol for 2 times; and (3) dissolving DNA by adding 100 mu L of sterilized ultrapure water, and using the DNA as a template. The amplified DNA fragments were sequenced and aligned by specific primer amplification (forward primer: GCAGTGCGTAGGAAGGAACT; reverse primer: GGTGTGCGTTGTTATACGCC), and the gene editing type of the created gene editing strain was confirmed (as shown in FIG. 1B).
(2) Gene editing mutant drought tolerance detection:
wild type B73-329 andZmNIP2a-d1、ZmNIP2a-d2the mutants are respectively planted in nutrition bowls, the weight of nutrition soil in each nutrition bowl is the same, and the mutants are cultivated in a greenhouse. After 10 days of growth, watering was stopped, drought treatment was performed, and drought phenotype during the treatment was observed. After 5 days of drought treatment, both gene-edited lines exhibited a more severe leaf wilting and curling phenotype, while no leaf curl was apparent in response to controls B73-329 (fig. 2A). The relative water content of the leaf of the plant at this time was examined (as follows), and the relative water content of the leaf of the two gene editing mutants was significantly lower than that of B73-329 (FIG. 2B). Active oxygen accumulation and oxidative damage are one of the important hazards of drought to plants. Detection of B73-329 and after drought treatmentZmNIP2a-d1、 ZmNIP2a-d2The content of Malondialdehyde (MDA) which is a membrane oxidation product in the mutant shows that the MDA content of two gene editing strain tissues after drought treatment is obviously higher than that of a control B73-329 (figure 2C), and the damage of the surface by drought stress is more serious.
The relative water content detection method comprises the following steps: the fresh weight of the plant tissue is weighed and recorded as W1, then the plant tissue is placed in an oven for full drying at 65 ℃, the dry weight is obtained by weighing, W2 is obtained by recording, and the relative water content of the plant tissue can be obtained by using a formula of [ relative water content= (W1-W2)/W1 x 100% ].
MDA (membrane oxidation product malondialdehyde) is measured by weighing 0.2 g experimental material into a centrifuge tube with steel balls in advance, immediately placing into a high-throughput tissue grinder after quick freezing by liquid nitrogen, vibrating and grinding for 1 min at 8,000 rpm, then adding 2 mL of 10% TCA (trichloroacetic acid) solution at 4 ℃ and 4,000 rpm, centrifuging for 10 min, sucking 1 mL supernatant, adding 1 mL of 0.5% TBA solution, uniformly mixing, reacting in a boiling water bath for 15 min, cooling, 4 ℃ and 10,000 rpm, centrifuging for 10 min, transferring 200 mu L of supernatant into an ELISA plate after centrifuging, measuring the absorbance at three positions of 532 nm, 600 nm and 450 nm in the ELISA meter, and calculating MDA content (mu mol/g) =MDA concentration (mu mol/L) multiplied by the volume of extract (mL)/weight of sample (g) according to the following formula.
(3) Transgenic overexpression genetic germplasm creation and expression detection:
using the cDNA of corn wild type B73-329 as a template, a specific primer was designed (primer sequence is as follows:
F: gcgcgccatttaaatactagtATGTCGACCAACTCGAGGTCCAACTC；
r catggtggatcccatactagtCACTTGGATGTGGTCGAGCTCGTCGTC, wherein the lower case letters are the joint sequences on the transgenic super-expression vector, the upper case letters are the primers in the coding sequence of the Zm00001d018037 gene), and PCR amplification and purification are carried out to recover amplified fragments. The amplified fragment recovered by purification was ligated with the transgenic overexpressing empty vector pcmcia 3300 digested with restriction enzymes (single step cloning ligation of novzan). The ligation product was transformed into E.coli competent cells and cultured overnight on a solid LB medium containing kanamycin, thereby selecting positive transformants. And extracting the successfully connected recombinant plasmid DNA and sequencing. And (3) transforming the recombinant vector into an agrobacterium EHA105 strain, submitting the constructed crop adversity adaptation of the Henan university province and improving a national key laboratory corn molecular breeding and gene editing platform to perform transgenic overexpression genetic germplasm creation work.
Recombinant Agrobacterium single colonies were inoculated into 2-3 mL liquid medium containing 100. Mu.g/mL kanamycin and 50. Mu.g/mL rifampicin, and shake-cultured overnight at 28 ℃. The next day was transferred to 50 mL liquid medium containing antibiotics for shake culture and resuspended to an OD600 of between 0.8-1.0 to obtain recombinant Agrobacterium suspension. And (3) infecting the maize immature embryo of B73-329 by adopting the obtained recombinant agrobacterium suspension, and then sequentially carrying out co-culture, resistance screening (the resistance screening adopts herbicide glufosinate), pre-differentiation, differentiation and rooting to obtain a T0 generation regenerated plant. Selfing the transgenic plants in the generation T0 to obtain generation T1 seeds, wherein the plants grown from the generation T1 seeds are generation T1 plants; the plants are selfed until T3 generation seeds are obtained, namely, the T3 generation seeds of the over-expression transgenic plants are obtained, the T3 generation lines obtained after the selfing of different T0 generation regenerated plants are respectively named as OE-1 and OE-2, and in the process, corn B73-329 is used as a control plant, namely, wild WT.
The genetic materials such as wild corn B73-329, transgene over-expression and the like are respectively 4 to 6 strains (OE-1 and OE-2), a few leaves are cut, and total RNA is extracted.
The extraction method of the specific corn RNA comprises the following steps: preparing a 1.5 mL RNase-free centrifuge tube with small steel balls, taking about 0.5 g fresh plant leaves, placing the fresh plant leaves in the centrifuge tube, covering the centrifuge tube with a cover, quick-freezing the fresh plant leaves in liquid nitrogen for 5-10 seconds, immediately placing the fresh plant leaves in a high-throughput tissue grinding instrument, and vibrating and grinding the fresh plant leaves at 8,000 rpm for 1 min to enable the leaves to fully crush powder (wearing masks and gloves in the process to prevent pollution of exogenous RNase); adding 1 mL pre-cooled Trizol on ice into the centrifuge tube, quickly mixing the mixture upside down, and standing the mixture at room temperature for 5 min; adding 200 μl of chloroform, shaking vigorously for 15 s, standing at room temperature for about 5 min, centrifuging at 1, 2000 rpm for 15 min at 4deg.C; separating the mixture into three layers after centrifugation, transferring the upper transparent water phase containing RNA into a 1.5 mL centrifuge tube without RNase, slowly sucking to avoid sucking into the middle and lower organic phase, and discarding the precipitate; adding isopropanol with the same volume, slightly reversing and uniformly mixing, standing for 10-20 min at room temperature, and precipitating RNA; centrifuging at 12,000 rpm and 4 ℃ for 10 min, obtaining RNA by a small amount of white precipitate at the bottom of the centrifuge tube, and discarding the supernatant; adding 1 mL of 75% ethanol, blowing, rinsing and precipitating, centrifuging at 12,000 rpm and 4 ℃ for 5 min, discarding the supernatant, and rinsing twice repeatedly; pouring the centrifuge tube into an ultra-clean bench after uncapping, naturally airing for 10 min, and fully volatilizing alcohol; 50 μl DEPC treated water was added to the centrifuge tube, RNA was fully dissolved, and the concentration was measured for reverse transcription to synthesize cDNA.
Preparation of cDNA: the extracted RNA was mixed in an RNase-free centrifuge tube using the Norpran HiScript 3 RT SuperMix for qPCR kit as follows
Genomic DNA removal system
| System component | Volume of |
| RNase-free ddH2O | to 16 μL |
| 4×gDNA wiper buffer | 4 μL |
| Template RNA | 1 pg-1μg |
After gently stirring and mixing, the mixture was reacted for 2 min at 42℃using a PCR instrument. After the completion of the reaction, 5X HIScript qRT SuperMix was directly added to the reaction tube.
Reverse transcription reaction system
| System component | Volume of |
| 5×HIScript qRT SuperMix | 4 μL |
| Reaction solution of the first step | 16 μL |
After mixing by gentle blowing, the mixture was reacted at 37℃for 15 minutes using a PCR instrument, and reacted at 85℃for 5 s. The product can be used for qPCR reaction immediately after measuring the concentration, or stored at-20 ℃.
Performing reverse transcription to obtain cDNA, performing PCR amplification with specific primer with the sequence of F5'-GTACGTACGTGTGCTAGCTAG-3' and R5'-GATGTCATGGATCTCGTTGTTG-3' to obtain amplified product, detecting the amplified product to obtain the relative expression of transgenic overexpressed strain gene,
the cDNA prepared by the reverse transcription kit is used as a template, and PCR amplification (F: 5'-GTACGTACGTGTGCTAGCTAG-3'; R: 5'-GATGTCATGGATCTCGTTGTTG-3') is performed by using specific primers to perform the identification work of the transcript level of the transgenic overexpressed strain.
The NIP2a transcripts in the overexpressing lines OE-1 and OE-2 were far higher than in the corresponding controls (B73-329), as shown in FIG. 2: the genetic materials such as transgene over-expression and the like reach about 20 times and 25 times higher than the relative expression quantity of wild genes of B73-329 respectively in corn strains OE-1 and OE-2 (figure 3).
Under the condition of the greenhouse, the genetic materials such as the earth cultivated wild corn (B73-329, namely wild WT), corn Zm00001d018037 transgene overexpression and the like reach 4 to 6 strains of corn strains (OE-1 and OE-2) respectively, and for comparison, corresponding gene editing mutant strains are planted at the same timeZmNIP2a-d1；ZmNIP2a-d2). Normally growing for 10 days, then stopping water supply, and performing drought treatment. After 5 days of drought treatment, compared with the wild nature (B73-329, wild nature WT) of the corresponding control material, the corn Zm00001d018037 gene edits nucleotide deletion mutationZmNIP2a-d1The method comprises the steps of carrying out a first treatment on the surface of the Coding sequence shift mutant of corn Zm00001d018037 geneZmNIP2a-d2) The sensitive phenotype was shown, drought tolerance was significantly reduced, while the transgenic overexpressed maize lines (OE-1 and OE-2) showed significantly increased drought tolerance (as shown in FIG. 4), and the above experiment was repeated 8 times with completely consistent results.
The studies of the inventors demonstrate that Zm00001d018037 gene improves drought stress response and promotes osmotic stress tolerance of plants. The functional identification of the gene under drought and osmotic stress of plants shows the innovation of the invention. These achievements have important theoretical and production significance for promoting drought stress tolerance of plants and the like.
Since osmotic stress is one of the most important aspects of drought stress, the present invention also investigated the expression of the maize Zm00001d018037 gene in yeast and promoting the growth of yeast under osmotic stress conditions.
Constructing a ZmNIP2a yeast expression fusion vector (pYES 2) vector system in which the ZmNIP2a gene is driven by GAL promoter and glucose is the promoter inhibitor, so that in a medium containing glucose,ZmNIP2agenes are not normally expressed, and in other glycogen media, such as in galactose-containing media,ZmNIP2acan be expressed, thereby causing the difference of growth vigor under the osmotic stress of yeast.
HandleZmNIP2aConstruction of the Gene coding sequence topYES2In the fusion vector formed, the constructed fusion vector is then transformed into a yeast (INVSV 1) strain. On glucose-containing medium, there was no significant difference in the vigor of sorbitol-treated and untreated transformants, whereas on sorbitol-supplemented galactose yeast medium, along with dilution of the yeast concentration gradient, the vigor of the yeast cells carrying the ZmNIP2a recombinant transformants was significantly better than that of the yeast cells transformed with the pYES2 empty vector, i.e. ZmNIP2a promoted yeast growth under osmotic stress caused by sorbitol. (as shown in fig. 5).
Since osmotic stress is one of the most important aspects of drought stress, the invention also researches the germination condition of the arabidopsis seeds under the osmotic stress condition promoted by the Zm00001d018037 gene of corn.
Method for creating transformation by adopting agrobacterium to infect arabidopsis thalianaZmNIP2aAnd (3) detecting the phenotype of the transgenic overexpressed Arabidopsis strain under osmotic stress. Normal arabidopsis seeds (wild type and transgenic overexpression) were inoculated on MS and ms+sorbitol medium, the phenotype of the seeds germinated for 6 days (a), and germination rate statistics at 4 days (B). The above experimental results show that overexpression in ZmNIP2a arabidopsis thaliana promotes germination under osmotic stress. "x" indicates that the difference is extremely significant.
Compared with wild type Arabidopsis thaliana (Col-0), the transgenic overexpressing Arabidopsis thaliana strain (OE-4, OE-5) exhibited enhanced tolerance to osmotic stress (MS+150 mM sorbitol) in the seed germination stage, and the germination rate of the transgenic overexpressing strain and the like was significantly higher than that of WT when grown for 4 days (FIG. 6).
The foregoing is merely a preferred embodiment of the invention, and it should be noted that modifications could be made by those skilled in the art without departing from the principles of the invention.
Claims (10)
1. The application of the corn gene Zm00001d018037 in improving the drought resistance of monocotyledonous crops.
2. The use according to claim 1, wherein the monocotyledonous crop is maize.
3. The use according to claim 1 or 2, in germplasm resource improvement.
4. The use according to claim 1 or 2, in genetic breeding.
5. Use according to claim 1 or 2, for the preparation of corresponding drought-resistant transgenic crops.
6. A method for improving the drought stress response of monocotyledonous crops, characterized in that the maize gene Zm00001d018037 according to claim 1 is integrated into monocotyledonous crop cells, tissues or organs and overexpressed.
7. The method of claim 6, wherein the monocot crop is maize.
8. A method for cultivating high drought resistant corn, comprising the steps of: transforming corn gene Zm00001d018037 into corn young embryo, co-culturing, resistance screening, callus induction, subculturing and differentiation to obtain T 0 Transgenic corn with stable T is obtained through continuous selfing and target gene PCR detection 3 And (3) obtaining the maize with the excessive expression of Zm00001d018037, namely the maize with high drought resistance, by using the generation homozygous transgenic line.
9. The use of the maize gene Zm00001d018037 for improving the osmotic stress tolerance of a yeast cell, characterized in that the maize gene Zm00001d018037 is expressed in the yeast and the osmotic stress tolerance of the yeast cell is improved.
10. The application of the corn gene Zm00001d018037 in improving the seed germination capacity of arabidopsis thaliana under the osmotic stress is characterized in that the corn gene Zm00001d018037 is expressed in arabidopsis thaliana, and the seed germination capacity of transgenic arabidopsis thaliana under the osmotic stress is improved.
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