CN103305529B - ZmWRKY50 gene improves the application of plant alumite - Google Patents

Abstract

The present invention relates to plant stress-resistance field, particularly, relate to the application that ZmWRKY50 gene improves the alumite of plant.The present invention is by the detection of RT-qPCR, and result proves that the expression of ZmWRKY50 gene is subject to AlCl ₃induction in various degree and suppression.Along with AlCl ₃the continuous increase of concentration, the expression of ZmWRKY50 gene is first raised in reduction.Work as AlCl ₃during concentration overrich, produce serious toxic action to plant, the expression of ZmWRKY50 gene also correspondingly receives suppression, illustrates at appropriate AlCl ₃in concentration range, the expression of ZmWRKY50 gene is induced, and just regulates and controls the acid-aluminum stress of plant, improves the antiacid aluminium ability of plant.

Description

Application of ZmWRKY50 gene to improve plant aluminum tolerance

technical field

The invention relates to the field of plant stress resistance, in particular to the application of the ZmWRKY50 gene to improve the aluminum tolerance of plants.

Background technique

As an important food, feed and economic ternary crop in my country, corn is affected by various adverse external environments such as pests, drought, salt damage, cold damage, and high temperature during its growth, thereby inhibiting its growth and development. Applying modern molecular biology techniques to excavate important stress-resistant genes and carry out genetic transformation or aggregate them with other excellent traits through molecular marker-assisted selection is one of the effective methods to obtain maize varieties with strong stress tolerance and improve maize yield. one. Functional research on stress-related genes is a prerequisite for the development and utilization of these genes. WRKY transcription factors play an important regulatory role in plant biotic stress, abiotic stress response and growth and development. WRKY transcription factors can participate in many different signaling pathways, and their functions are diverse. Therefore, the WRKY gene and its regulated target genes are potentially important stress resistance genes.

During the growth process, plants will be affected by various adverse external environments such as pests and diseases, drought, salt damage, cold damage, high temperature, soil pH, and lack of mineral nutrients, which inhibit the growth and development of plants and lead to regional limitations in plant distribution. sex. At the same time, plants have formed a series of mechanisms to adapt to and resist various adversity stresses in the long-term evolution process. When plants are under adversity stress, this extracellular stimulus signal enters the cell through cascade signal transduction, activates transcription factors, promotes their binding to downstream target genes and promotes the expression of the gene at the transcriptional level, thus responding to the plant's response reaction. At present, ten large transcription factor families have been discovered in higher plants. The WRKY gene family is one of them. In addition to regulating the plant hormone signal transduction pathway, it is also widely involved in plant biotic and A series of physiological activities such as biosynthesis, organ development and plant aging.

Due to the limited germplasm resources such as resistance to stress and disease and its genetic mechanism is very complicated, it is difficult to organically combine the traits of disease resistance and stress resistance with the yield and quality of crops in the process of conventional breeding to breed resistant varieties that meet the needs of agricultural production crops. The rapid development of modern molecular biology technology provides a good opportunity to solve this problem. Therefore, applying modern molecular biology techniques to excavate important plant disease-resistant and stress-resistant genes and genetically transform them is one of the effective methods to obtain crop varieties with strong stress tolerance and to solve and improve agricultural yields. WRKY transcription factor is one of the ten largest transcription factor families in higher plants. WRKY transcription factors participate in various stress responses of plants, and play an important regulatory role in the growth and development of plants. Therefore, WRKY genes and their regulated target genes are potentially important stress resistance genes. So far, there are very few reports on the involvement of WRKY transcription factors in plant aluminum resistance. The research on the WRKY transcription factor family in maize under abiotic stress is still in its infancy. At present, maize WRKY gene has been found to be related to maize disease and salt resistance, but there is no report related to antacid aluminum resistance.

Contents of the invention

The purpose of the present invention is to provide the application of maize ZmWRKY50 gene for antacid aluminum.

Another object of the present invention is to provide a method for providing plant antacid aluminum.

According to the technical scheme of the present invention, first clone the ZmWRKY50 gene from the corn inbred line, and its nucleotide sequence is as shown in SEQ IN No.1:

AAACACCTCCCAACCCAATCTCCCCAGAGAGAGAGAGCCCCAAGCCAAGATCCGCGCGAAGCAAGTCACCCGGCGAAGCACCGGCTCCCATGGCCGTGGACCTGATGGGCTGCTACGCCCCGCGCCGCGCCAACGACCAGCTCGCCATCCAGGAGGCGGCGGCGGCCGGGCTCCGCAACCTGGAGCTGCTGGTGACGTCCCTGTCCACGCAGGCCGCCGCGCCGCACAGAGCCGCTGATCAGCCGTTCGGCGAGATCGCCGGCCAGGCCGTCTCCAAGTTCCGCAAGGTCATCTCCATCCTCGACCGCACGGGGCACGCCCGCTTCCGCCGCGGGCCCGTCGAGCCGCCGCCGCCGACGCCGCCGCCGCCTCCTGTCGTCCCCGGTCCTGCCCCCCTGGCGGCCGTCAGCGTGGCGCAGCCGCCGCAGAGCCTGACGCTGGACTTCACGAAGCCGAACCTGGCCGTGTCGGCCGCCACGTCCGTCACCTCCACGTCCTTCTTCTCGTCGGTCACGGCCGGCGAGGGCAGCGTCTCCAAGGGCCGGAGCCTCATGTCCTCCGGGAAGCCGCCGCTGTCTGGCCACAAGCGGAAGCCCTGCGCCGGCGCCCACTCCGAGGCCACCACCAACGGCAGCCGGTGCCACTGCTCCAAGAGAAGGAAGAACCGCGTGAAGAGGAGCATCAGAGTGCCGGCGATCAGCTCGAAGGTCGCCGACATCCCTTCGGACGAGTACTCGTGGAGGAAGTACGGCCAGAAGCCCATCAAGGGCTCCCCTTACCCACGGGGCTACTACAAGTGCAGCACTGTGCGGGGATGCCCGGCGAGGAAGCACGTGGAGCGCGCCACCGACGACCCGGCCATGCTGGTGGTGACGTACGAGGGCGAGCACCGCCACACGCCGGGCGCGGTCCAGGGGCCGAGCCCCCTGGCGACCGCGTCGCCGGTGCCCGTCGCCGTCTCCGCCGGCAACGGGCTCGTTGTCTAGTCTACTAAAAGCTA GGATTAGCTTCTCGTCTTCTTTGTTTTTTTTTTGTTTGAGCTGATGTCCGTGTAAACAAGGAAGAAGGTTGTAGAAAGAGGGAG

Furthermore, the present invention uses RT-qPCR detection to prove that the expression of the ZmWRKY50 gene is induced and inhibited by AlCl ₃ to varying degrees. With the increasing concentration of AlCl ₃ , the expression of ZmWRKY50 gene was first up-regulated and then decreased. When the concentration _of AlCl3 is too high, it will cause serious poisoning to plants, and the expression of ZmWRKY50 gene will be inhibited accordingly, indicating that within the range of appropriate AlCl3 concentration, the expression of _ZmWRKY50 gene is induced, and the acidity of plants Aluminum stress is positively regulated to improve the ability of plants to resist aluminum.

Through genetic transformation studies, it was found that overexpression of ZmWRKY50 gene can improve the aluminum tolerance of plant roots, and the roots of 35S::ZmWRKY50 plants can promote their growth when they are stressed by a specific concentration of AlCl ₃ .

It can be seen that the present invention provides the application of the ZmWRKY50 gene to improve the aluminum tolerance of plant roots.

Description of drawings

Figure 1 shows the amplification of the ZmWRKY50 gene in Figure 1, M: DL2000; 1: PCR product of ZmWRKY50;

Fig. 2 shows the influence of 60uM AlCl ₃ on ZmWRKY50 gene expression;

Fig. 3 shows the expression situation of the tissue site of ZmWRKY50 gene;

Figure 4 shows the digestion and PCR detection of the plant expression vector pCBP-ZmWRKY50, M: DL2000; 1: XbaI/SacI double digestion of pMD19-T-ZmWRKY50; 2: XbaI/SacI double digestion of pCBP; 3-4: pCBP- PCR detection of ZmWRKY50; 5: XbaI/SacI double enzyme digestion detection of pCBP-ZmWRKY50;

Figure 5 shows the PCR detection of pCBP-ZmWRKY50 transformed into Agrobacterium, M: DL2000; 1-3: the PCR detection of pCBP-ZmWRKY50;

Figure 6 shows the screening and detection of 35S::ZmWRKY50 plants, A: Basta screening of T0 generation plants; B1: ZmWRKY50 positive control; B2-12: part of ZmWRKY50 transgenic Arabidopsis; B13: ddH2O negative control; B14: wild type Arabidopsis negative control;

Figure 7 shows the effect of acid aluminum stress on the growth length of 35S::ZmWRKY50 roots;

Figure 8 shows the effect of acid aluminum stress on the net growth of 35S::ZmWRKY50 roots.

Detailed ways

Example 1

1. Acquisition and cloning of ZmWRKY50 gene sequence

In the maizesequence database, the cDNA and protein sequences of the ZmWRKY50 gene were obtained.

The cDNA sequence of ZmWRKY50 gene was analyzed, and the primer W50F/R was designed. W50F: tatctagagacaaacacctcccaaccc; W50R: atgagctcctccctctttctacaaccttcttc;

Using the cDNA of maize inbred line 178 as a template, the ZmWRKY50 gene was specifically amplified.

The reaction system is:

Table 1 PCR reaction system

The PCR reaction program is: 95°C for 3min; 95°C for 30sec, 60°C for 30sec, 72°C for 1.5min 35cycles; 72°C for 5min. The PCR product was detected by 1.0% agarose gel electrophoresis, and the target band was recovered using the agarose gel recovery kit of Omega Company.

Amplification of the ZmWRKY50 gene

Use the specific primer W50F/R of ZmWRKY50 to amplify the cDNA of these two genes from maize inbred line 178, obtain the fragment of 1087bp of ZmWRKY50 gene (as Fig. The sequence homology of the inbred line B73 was 99.63%, indicating that the amplified fragments were the ZmWRKY50 gene.

2. Expression analysis of ZmWRKY50 gene under acid aluminum stress

2-1. Different acid aluminum stress treatments of maize inbred line materials

Select plump corn inbred line 178 seeds and sterilize them with 75% ethanol for 2 minutes, soak in 1℅ NaClO solution for 8-10 minutes and shake constantly during the disinfection, and then rinse with deionized water for 3-5 times until the residual NaClO is rinsed clean. Then soak in deionized water for 4 hours, and finally germinate with filter paper in the dark at 28°C. After about 2-3 days, transfer the germinated corn to a floating plate, and culture it with Hoagland nutrient solution under the conditions of light (16h)/darkness (8h) at 28°C for hydroponics, change the culture medium twice a week, and wait until the seedlings grow to Stress treatment was carried out at the three-leaf stage.

2-2. Acid aluminum stress:

(1) Cultivate the seedlings with deionized water containing 200 μM CaCl for 24 hours, and adjust the pH to 4.2;

(2) Treat the seedlings with a solution containing 0 μM, 30 μM, 60 μM, 120 μM, 240 μM, 500 μM AlCl ₃ for 0 h, 6 h, 12 h, 24 h, 30 h. Parts taken: whole plant, 0h treatment as control.

Maize inbred line 178 at the seedling stage was treated with 60uM AlCl ₃ for 0h, 1h, 6h, 12h, 24h, 30h. RT-qPCR analysis found that ZmWRKY50 gene expression was significantly induced by AlCl ₃ stress. During the 1-hour treatment period, the expression level was up-regulated by about 2 times. Especially when the treatment time was 6 hours, the expression of ZmWRKY50 gene showed a peak, and compared with the control, the expression intensity increased by about 6.8 times, as shown in Figure 7 . With the extension of AlCl ₃ stress time, the expression of ZmWRKY50 gene hardly changed. Therefore, ZmWRKY50 gene is induced and expressed by AlCl ₃ stress, and is strongly induced in a short period of time, which may positively regulate the response of plants to acid aluminum stress (see Figure 2).

2-3. Extraction of corn total RNA

For the removal of DNA in RNA samples and reverse transcription of RNA, use the TakaRa kit to remove residual genomic DNA in RNA. The reaction system (1) is as follows:

2-4. Analysis of spatiotemporal expression pattern of ZmWRKY50 gene

2-1 The cDNA of the treated sample was used as a template, ACT2 and GAPDH were used as internal reference genes, and the relative quantitative analysis of the expression of the ZmWRKY50 gene under the acid-aluminum stress condition was carried out by fluorescent quantitative PCR technology.

First, these genes were cloned, and the reaction parameters were set as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 sec, Tm (see table) annealing for 30 sec, extension at 72°C for 30 sec for 35 cycles; final extension at 72°C for 5 min. PCR amplification products of each gene were detected by 1.5% agarose gel electrophoresis. The target fragment is recovered from the gel and connected to the T-vector for sequencing to check whether the fragment amplified by the primer is the target gene.

Preparation of standard curve: recover and purify the PCR product of the correctly sequenced gene, perform 10-fold gradient dilution with ddH ₂ O, and take RNA samples with a concentration of 10 ^-2 to 10 ^-9 as the standard for preparing the standard curve. The corresponding amplification efficiency E=(10 ^[-1/slope] -1)×100% was obtained for each gene through the slope of the corresponding standard curve. The reaction program is: 95°C for 30s; 95°C for 5s, Tm (primer optimum annealing temperature) for 30s, 40 cycles; 95°C for 10s; 65°C for 5s, draw a melting curve from 65°C to 95°C. After the standard curve was drawn, the loading experiment of the target gene was carried out. Using ^{the 2-△△Ct} relative quantitative formula, where △△Ct=(Ct _Target -Ct _Actin ) _x- (Ct _Target -Ct _Actin ) ₀ , the expression level of the ZmWRKY50 gene whose treatment time was 0h was set as 1, as As a control, the relative expression of the ZmWRKY50 gene in different treatment time periods was calculated quantitatively. Three repetitions were set for each sample. The PCR reaction system is as follows:

Table 2 RT-qPCR amplification system

The reaction procedure is:

95°C for 30s; 95°C for 5s, Tm (primer optimum annealing temperature) for 30s, 40cycles; 95°C for 10s; 65°C for 5s, draw the melting curve from 65°C to 95°C. The sequences of primers for quantitative PCR of the genes used are shown in Table 5:

Table 3 Fluorescent real-time quantitative PCR primers

60uM AlCl ₃ treated maize inbred line 178 at the seedling stage for 0h and 6h. By RT-qPCR analysis, it was found that the expression of ZmWRKY50 gene in roots, stems and leaves was significantly induced by AlCl ₃ stress. Among them, the expression was the strongest in the roots, indicating that the ZmWRKY50 gene was specifically expressed in the roots of maize (the results are shown in Figure 3).

2-5. Research on genetic transformation of ZmWRKY50 gene

Construction of pCBP-ZmWRKY50 overexpression vector

Since the ZmWRKY50 gene is amplified with KOD high-fidelity enzyme to form a blunt end, it needs to add a PolyA tail to the 3' end in order to connect with 19-T. The reaction system is: PCR product 7.3 μL; 10×Buffer 1 μL; dNTP 1.5 μL; rTaq0 .2 μL. The reaction program is: 72°C for 30 min, 12°C forever.

The above fragments were respectively ligated into pMD19-T vector. The ligation reaction system was: 4.5 μL of PCR recovered product, 0.5 μL of pMD19-T Vector, and 5 μL of Solution I. After connecting at 16°C for 3 hours, transform Escherichia coli DH5α strain, and after culturing overnight, pick positive single clones and send them to Invitrogen for sequencing.

A small plasmid extraction kit was used to extract the positive plasmid pMD19-T-ZmWRKY50. The pMD19-T-ZmWRKY50 and the plant expression vector pCBP were double-digested with restriction enzymes XbaI and SacI respectively; the target fragment was recovered after digestion at 37°C for 3 hours. The digestion system is as follows: 5 μl of plasmid DNA, 2 μl of 10× digestion buffer, 0.5 μl of XbaI, 0.5 μl of SacI, and 12 μl of ddH ₂ O.

The recovered ZmWRKY50 digested fragments were ligated with the corresponding pCBP digested fragments respectively. The ligation system was: 0.5 μL of vector DNA, 4.5 μL of target fragments, and 5 μL of Solution I. Ligation at 16°C for 3 hours. The recombinant plasmid was transformed into Escherichia coli DH5α competent cells, and the transformation method was as follows: (1) Take 200 μL of Escherichia coli competent cells and place them on ice until thawed. (2) Add 0.5 μL of positive plasmid DNA to competent cells, stir gently and place on ice for 30 minutes. (3) Water bath at 42°C for 90 seconds, take it out and immediately insert it into ice, and let it stand for 2 minutes. (4) Under sterile conditions, add 800-1000 μL of fresh anti-LB liquid medium, culture at 37° C. with shaking at 200 r/min for about 1 hour. (5) Pipette 200 μL of the transformed bacteria solution and evenly spread it on the LB (Kan ⁺ ) plate, and culture it upside down at 37° C. for 12-16 hours. After overnight culture, single clones were picked for colony PCR detection and plasmid digestion detection. Finally, the plasmid extraction kit was used to extract the positive plasmid pCBP-ZmWRKY50 for corresponding enzyme digestion verification, and the results obtained bands of about 1100bp, indicating that the vector was constructed correctly (Figure 4).

Transformation of Arabidopsis with pCBP-ZmWRKY50 Overexpression Vector

(1) Agrobacterium-mediated transformation of Arabidopsis inflorescence

Take 200 μL of Agrobacterium EHA105 competent cells, add 1 μL of carrier plasmid DNA after thawing, stir gently, mix well, and ice-bath for 30 minutes to carry out Agrobacterium transformation, then quick-freeze in liquid nitrogen for 1 minute, and after 5 minutes in 37°C water bath, add 1 mL of no Antibiotic fresh YEB medium. Finally, 28 ℃, 150r/min shaking culture 4h. Pipette 300 μL of the bacterial solution and spread it on a YEB (containing 50 μg/mL kanamycin and 50 μg/mL rifampicin) culture plate, culture it upside down at 28°C for about 48 hours, and pick a single colony for bacterial liquid PCR identification. Positive colonies were inoculated in YEB liquid medium containing 50 μg/mL kanamycin and 50 μg/mL rifampicin. Expand the culture to OD600 of about 1.2-1.6 under the condition of 28°C and 200r/min. Finally, 4000r/min was centrifuged for 10min to collect the bacteria, and the bacteria were diluted to OD600 value of 1.0 with 10% sucrose water, and SilwetL-77 with a volume ratio of 1/10,000 was added, and the Arabidopsis inflorescence was soaked in the bacteria solution for about After 1 min, the infected Arabidopsis thaliana was cultured in the dark for 16 h and then returned to normal culture. Figure 5 shows that the pCBP-ZmWRKY50 overexpression vector has been successfully transformed into Agrobacterium.

(2) Molecular detection of 35S::ZmWRKY50 Arabidopsis plants

As shown in Figure 6, put the transgenic Arabidopsis seeds of T ₀ generation in a 1.5mL centrifuge tube, add 1mL of 75% alcohol to soak for 1min, pour off the alcohol, then add 1mL of 0.1% sodium hypochlorite to sterilize for 10min, and use sterile The seeds were washed with ddH ₂ O for 6 times, the supernatant was discarded, and finally the seeds were coated with 0.1% sterile agar water on the selection medium containing 15 mg/LBasta for selection. After 7 days, the Arabidopsis seedlings that survived the screening were transplanted into the substrate soil for growth. Or sow the transgenic Arabidopsis seeds of the T ₀ generation in nutrient soil, and spray the surface of the Arabidopsis leaves with 15 mg/L of Basta solution when they have four leaves for screening. When the plant grows to 8-10 leaves, use the kit to extract the genomic DNA of the leaf, identify the positive transgenic plant by PCR, obtain the target fragment of about 440bp, and harvest it as a single plant (T ₁ generation).

35S::ZmWRKY50 Arabidopsis thaliana selected 35SW50F/R primers for genome transgene PCR detection. The partial sequence of the 35S promoter gene and the internal partial sequence of the target gene are selected as primer pairs for detection primers. The length of the detected ZmWRKY50 gene product is about 446bp. After the band with the same size as the target fragment was obtained by PCR, the band was recovered and ligated with T carrier for sequencing.

Table 4 Information of detection primers

Analysis of Aluminum Tolerance of 2-635S::ZmWRKY50 Arabidopsis Plants

(1) According to the above screening method for transgenic Arabidopsis plants, screen 35S::ZmWRKY50 plants and obtain homozygous T ₃ generation seeds. The total RNA of T3 generation homozygous strains was extracted and used as a template, and qW50F/ _R was used as primers for semi-quantitative PCR to detect the expression of ZmWRKY50 gene in different homozygous strains. The two lines with the highest expression levels were selected as representatives to analyze the aluminum tolerance phenotype of 35S::ZmWRKY50 plants.

(2) Effect of AlCl ₃ stress on root growth of 35S::ZmWRKY50 plants

The homozygous T ₃ generation seeds of 35S::ZmWRKY50 plants grown on 1/2MS medium for 7 days were transplanted into 1/2MS medium with AlCl ₃ concentrations of 0, 50uM, 100uM, 150uM, pH 4.2, and wild-type Arabidopsis was used as a control, and the effect of AlCl ₃ on root growth length of transgenic Arabidopsis plants was observed after 10 days. The T ₃ seeds of the two pure line strains L3 and L9 with the highest expression levels and the wild-type seeds were sterilized and planted on 1/2 MS medium for growth. 7d to measure the root length of the plant, and then transplant the L3 and L9 seedlings in the 1/2MS medium with AlCl ₃ concentrations of 0, 50uM, 100uM, 150uM, and pH 4.2, and wild-type Arabidopsis as a control. After 7 days of cultivation Measure its root length again, calculate the root growth length of L3 and L9 and WT plant.

The T ₃ seeds of the two pure line strains L3 and L9 with the highest expression levels and the wild-type seeds were sterilized and then grown on 1/2 MS medium. After 7 days, the L3 and L9 seedlings were transplanted in the 1/2MS medium with AlCl ₃ concentrations of 0, 50uM, 100uM, 150uM, and pH 4.2. The wild-type Arabidopsis was used as the control. After 10 days, the AlCl ₃ effect on the transgenic Arabidopsis The effect of root growth length of mustard plants is shown in Figure 7. With the increase of AlCl ₃ concentration, the root length of WT plants gradually shortened, and the difference in root length between WT and 35S::ZmWRKY50 plants became more and more obvious, indicating that AlCl ₃ stresses the growth of WT plants and inhibits the growth and elongation of roots. However, the increase of AlCl ₃ concentration did not significantly inhibit the growth of 35S::ZmWRKY50 roots, and when the AlCl ₃ concentration was 150uM, the roots of 35S::ZmWRKY50 plants were not inhibited but became longer and thicker, and the lateral roots increased , showing a vigorous growth trend. It shows that 35S::ZmWRKY50 plants have a certain ability of tolerance to aluminum, and the ZmWRKY50 gene positively regulates the aluminum stress of plants.

The concentration of AlCl ₃ was 0, 50uM, 100uM, 150uM, respectively, and the 1/2MS medium with pH 4.2 was stressed for 7 days, and the net semiminal rootlength (NSRL) of L3 and L9 transgenic lines and WT plants =FRSL-ISRL) as shown in FIG. 8 . With the increase _of AlCl3 concentration, the root net growth length of WT gradually decreased. When the concentration of AlCl ₃ was 0uM, there was no significant difference in the net root growth length of L3, L9 and WT plants. When the concentration of AlCl ₃ was 50uM, the root growth of WT decreased more than that of L3 and L9, and the net root growth of L3 and L9 was significantly higher than that of WT plants. When treated with 100uM AlCl ₃ , the root net growth of WT and L3, L9 plants continued to decrease, but the root net growth of L3, L9 plants was significantly higher than that of WT plants. When the concentration of AlCl ₃ was 150uM, the net root growth of WT continued to decrease, but the net root growth of L3 and L9 plants increased significantly, and the difference between the two reached the largest. It shows that under AlCl ₃ stress conditions, the roots of L9 and L3 plants are more suitable for growth than the roots of WT plants, and the plants have tolerance to aluminum.

According to the detection of RT-qPCR in the present invention, the result proves that the expression of ZmWRKY50 gene is induced and inhibited by AlCl ₃ in different degrees. With the increasing concentration of AlCl ₃ , the expression of ZmWRKY50 gene was first up-regulated and then decreased. When the concentration _of AlCl3 is too high, it will cause serious poisoning to plants, and the expression of ZmWRKY50 gene will be inhibited accordingly, indicating that within the range of appropriate AlCl3 concentration, the expression of _ZmWRKY50 gene is induced, and the acidity of plants Aluminum stress is positively regulated to improve the ability of plants to resist aluminum.

Through genetic transformation studies, it was found that overexpression of ZmWRKY50 gene can improve the aluminum tolerance of plant roots, and the roots of 35S::ZmWRKY50 plants can promote their growth when they are stressed by a specific concentration of AlCl ₃ .

Claims (1)

1.ZmWRKY50 gene improves the application of the alumite of plant root, and wherein, the nucleotide sequence of described gene is as shown in SEQ IN No.1, and described plant is corn or Arabidopis thaliana.