阅读说明：本技术 一种植物启动子及其应用 (Plant promoter and application thereof ) 是由 彭巍 傅明辉 邓志威 陆梓晴 周淑贤 于 2020-06-24 设计创作，主要内容包括：本申请属于基因工程的技术领域,尤其涉及一种植物启动子及其应用。本申请提供了一种植物启动子,包括脱落酸响应元件、诱导厌氧响应元件、光响应元件、启动子和增强子区域作用元件、赤霉素响应元件、调控分生组织特异活性元件、茉莉酸响应元件、乙烯响应元件、干旱、高盐、低温响应元件、抗脱落酸响应元件、转录核心启动子元件、GGTGC功能区和转录起始点。本申请公开的新型植物启动子,该启动子能够在植物中启动目的基因的高效表达。(The application belongs to the technical field of genetic engineering, and particularly relates to a plant promoter and application thereof. The application provides a plant promoter, which comprises abscisic acid response elements, induced anaerobic response elements, light response elements, promoter and enhancer region action elements, gibberellin response elements, regulation and control meristem specific activity elements, jasmonic acid response elements, ethylene response elements, drought, high-salt, low-temperature response elements, abscisic acid response elements, transcription core promoter elements, GGTGC functional regions and transcription starting points. Disclosed herein are novel plant promoters capable of promoting efficient expression of a gene of interest in a plant.)

1. A plant promoter comprising an abscisic acid-responsive element, an induced anaerobic-responsive element, a light-responsive element, a promoter-and-enhancer region-acting element, a gibberellin-responsive element, a regulatory meristem-specific active element, a jasmonic acid-responsive element, an ethylene-responsive element, drought, high-salinity, low-temperature-responsive element, an anti-abscisic acid-responsive element, a transcriptional core promoter element, a GGTGC functional region, and a transcription initiation point.

2. The plant promoter of claim 1, wherein the abscisic acid-responsive element is selected from the group consisting of AAGAA-motif and ABRE;

the induced anaerobic response element is selected from ARE;

the photoresponsive element is selected from Box 4, G-Box and TCCC-motif;

the promoter and enhancer region acting elements are selected from CAAT-box;

the gibberellin-responsive element is selected from CARE;

the element for regulating and controlling the meristem-specific activity is selected from CCGTCC-box;

the jasmonic acid response element is selected from the group consisting of CGTCA-motif and TGACG-motif;

the ethylene responsive element is selected from ERE;

the drought, high salinity, low temperature responsive element is selected from MYB;

the anti-abscisic acid-responsive element is selected from MYC;

the transcription core promoter element is selected from the group consisting of TATA-box;

the transcription initiation point is selected from TSSP.

3. The plant promoter according to claim 1, having any one of the nucleotide sequences shown in (i), (ii) or (iii):

(I) is shown as SEQ ID NO. 1;

(II) a sequence having at least 70% homology with the sequence shown as SEQ ID No. 1;

(III) a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more nucleotides in the sequence shown in SEQ ID NO. 1.

4. The plant promoter according to claim 1, having any one of the nucleotide sequences shown in (iv), (v) or (vi):

(IV) is shown as SEQ ID NO. 2;

(V) a sequence having at least 60% homology with the sequence shown as SEQ ID No. 2;

(VI) a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more nucleotides in the sequence shown in SEQ ID NO. 2.

5. The plant promoter according to claim 1, having any one of the nucleotide sequences shown in (VII), (VIII) or (IX):

(VII) shown as SEQ ID NO. 3;

(VIII) a sequence having at least 50% homology to the sequence shown as SEQ ID No. 3;

(IX) a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more nucleotides in the sequence shown in SEQ ID NO. 3.

6. The plant promoter according to any one of claims 3 to 5, wherein the substitution is of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65 nucleotides.

7. Use of a plant promoter according to any one of claims 1 to 6 for promoting expression of a gene of interest in a plant.

8. Use according to claim 7, wherein the plant is selected from the group consisting of duckweed, tobacco and Arabidopsis thaliana.

9. The use according to claim 7, wherein the plant promoter is specifically: the plant promoter is used for promoting the specific expression of a target gene in the root, stem or leaf tissue of a plant.

10. An expression vector comprising the plant promoter according to any one of claims 1 to 6.

Technical Field

The application belongs to the technical field of genetic engineering, and particularly relates to a plant promoter and application thereof.

Background

With the vigorous implementation of national construction and the rapid development of industrialization process, more and more water bodies such as rivers and lakes and the like generate serious eutrophication phenomena. Since long ago, people utilize aquatic plants to purify eutrophic water, which is a universal, economical and efficient method. However, large-scale aquatic plants such as water hyacinth are easy to cause channel blockage, difficult to salvage and low in recovery value, and the random growth of the water hyacinth causes secondary pollution and other environmental problems, the aquatic plants have very wide development prospect in purifying eutrophic water, and the duckweed is expected to solve the dilemma.

The duckweed is called Duck grass and is a perennial herb, can live in lakes and rivers all the year round, is distributed in all parts of the world, and is widely distributed in tropical and temperate freshwater lakes and part of estuaries near the sea. Spirodela polyrhiza is a water surface floating plant of the genus Spirodela of the family Spirodelaceae, and has the characteristics of strong growth adaptability, good purification of eutrophic water body, easy salvage and recovery, and the like. The area of the duckweed water purification body is very wide, and most of organic sewage can be almost absorbed, such as: domestic sewage, human and animal excrement waste water, and waste liquid and waste material discharged from processing factories such as food and the like. In addition, the duckweed can absorb various heavy metal elements, has great potential as biological energy and has good medicinal value.

Therefore, the production of transgenic duckweed by genetic engineering technology is widely concerned, the plant promoter is the basis for constructing transgenic plants, the plant promoter is a DNA sequence which is positioned at the upstream of the 5' end of a gene initiation codon (ATG) and can be accurately combined with RNA polymerase, the direction and the efficiency of gene transcription are determined, and the plant promoter is the core for regulating and controlling the gene transcription. However, only a few promoters have been identified and widely used in duckweed.

Disclosure of Invention

In view of the above, the present application develops a novel plant promoter capable of promoting efficient expression of a target gene in a plant.

In a first aspect, the present application provides a plant promoter comprising an abscisic acid-responsive element, an induced anaerobic-responsive element, a light-responsive element, promoter and enhancer region-acting elements, gibberellin-responsive elements, regulatory meristem-specific active elements, jasmonic-responsive elements, ethylene-responsive elements, drought, high-salinity, low-temperature-responsive elements, anti-abscisic acid-responsive elements, transcriptional core promoter elements, GGTGC functional regions, and a transcriptional initiation point.

Preferably, the abscisic acid-responsive element is selected from the group consisting of AAGAA-motif and ABRE;

the induced anaerobic response element is selected from ARE;

the photoresponsive element is selected from Box 4, G-Box and TCCC-motif;

the promoter and enhancer region acting elements are selected from CAAT-box;

the gibberellin-responsive element is selected from CARE;

the element for regulating and controlling the meristem-specific activity is selected from CCGTCC-box;

the jasmonic acid response element is selected from the group consisting of CGTCA-motif and TGACG-motif;

the ethylene responsive element is selected from ERE;

the drought, high salinity, low temperature responsive element is selected from MYB;

the anti-abscisic acid-responsive element is selected from MYC;

the transcription core promoter element is selected from the group consisting of TATA-box;

the transcription initiation point is selected from TSSP.

It is noted that the nucleotide sequence of the GGTGC functional region is GGTGC.

Preferably, it has any one of the nucleotide sequences shown in (I), (II) or (III):

(I) is shown as SEQ ID NO. 1;

(II) a sequence having at least 70% homology with the sequence shown as SEQ ID No. 1;

(III) a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more nucleotides in the sequence shown in SEQ ID NO. 1.

The plant promoter of the present application can be obtained by artificial synthesis or by PCR. Specifically, the sequence of the first plant promoter disclosed by the application is shown in SEQ ID No.1 and is named as SpPHT1 promoter 3, and the nucleotide sequence of the SpPHT1 promoter 3 is as follows: GGGCACTCGTTGCTCCATCTCGGCTAATTTAACGAAATAATTGGAATGAGAGAGATGCTAAAAAAGGGGTCTTCCGTTTGGATAGTGATGCATCCCTCTGTCTCTCTCTCCTCCGGTAGGGCCTTCAAGGTTTTCGTTGAAACTCATCGGAAGATCAGTCTAAAGTTAAGGCAAGTGTCGTGTCGTCTTTCTCTCTCACGTTCCTGGACCCGTGATCGTTTGATTCCAGGAACTTTGGTGATCGATTGATTCTTGAGAGTCCAAAAATCGGTCAACGACTGCGAAATATTTTGAGCGGATTTTCATGTGCATCAATTTAAAATTGATATTTATTTATTTTGTGCTGTCTGGGCTTTTCAATTTCTTCAGAGAAATCCTGGGATCCTCTTCTTTCACCCTCGGATCTCATCCTCTGGACCTTCCATCTCTCTGGTTTCTTCCTCTGTAATTCAACTTATTATATGTACACACCGTCCTCTAAGATTTTGTGGATTCTTTACTTTGCAGGATCTGAAGGAGATTAAAAGGGGGTTAGACCATGGCAAGAGATCAGCTTCAGGTCCTCACCGCGCTAGATGTCGCCAAGACGCAGTGGTATCACTTCACGGCGATCGTGATCGCCGGCATGGGGTTCTTTACCGACGCCTACGACC are provided.

Specifically, the method for obtaining the plant promoter of SEQ ID NO.1 by the PCR method comprises the following steps: SpPHT1 promoter 3 (SpPHT1 gene promoter sequence, SEQ ID NO.1) is amplified by taking total duckweed genome DNA as a template. The PCR reaction system is as follows: DNA 2. mu.L, 10 XPCR Buffer 2.5. mu.L, SpPHT1-Q1-F (SEQ ID No.4) 1. mu.L, dNTP (10mM each) 1. mu.L, SpPHT1-Q1-R (SEQ ID No.5) 1. mu.L, LA Taq 0.5. mu.L, ddH₂O17. mu.L. PCR amplification conditions: pre-denaturation at 95 ℃ for 3 min; denaturation at 95 ℃ for 30s, annealing at 63.1 ℃ for 30s, extension at 72 ℃ for 1min, and 30 cycles; extension at 72 ℃ for 5 min. Wherein: the nucleotide sequence of SpPHT-Q1-F (SEQ ID No.4) is as follows: 5'-CCCAAGCTTGGGCACTCGTTGCTCCAT-3' (with Hind III cleavage sites at the 5 ' end); the nucleotide sequence of SpPHT1-Q1-R (SEQ ID No.5) is as follows: 5'-GCTCTAGAGGTCGTAGGCGTCGGTAA-3' (with an XbaI cleavage site at the 5 ' end).

Preferably, the nucleotide sequence has any one of the nucleotide sequences shown In (IV), (V) or (VI):

(IV) is shown as SEQ ID NO. 2;

(V) a sequence having at least 60% homology to the sequence shown as SEQ ID No. 2;

(VI) a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more nucleotides in the sequence shown in SEQ ID NO. 2.

The plant promoter of the present application can be obtained by artificial synthesis or by PCR. Specifically, the sequence of the second plant promoter of the present application is shown in SEQ ID No.2 and is named SpPHT1 promoter 2, the 5' end of SpPHT1 promoter 2 has a plurality of action elements more than SpPHT1 promoter 3, the nucleotide sequence of SpPHT1 promoter 2 is: TTGTGGTCGTGCGTGTGACGCTATCTAGTTAATTTTTTCAATCCATGCCGGATCATTTTTTTTTTCTAGCATTAGATAGCGGATTGGGGCACTCGTTGCTCCATCTCGGCTAATTTAACGAAATAATTGGAATGAGAGAGATGCTAAAAAAGGGGTCTTCCGTTTGGATAGTGATGCATCCCTCTGTCTCTCTCTCCTCCGGTAGGGCCTTCAAGGTTTTCGTTGAAACTCATCGGAAGATCAGTCTAAAGTTAAGGCAAGTGTCGTGTCGTCTTTCTCTCTCACGTTCCTGGACCCGTGATCGTTTGATTCCAGGAACTTTGGTGATCGATTGATTCTTGAGAGTCCAAAAATCGGTCAACGACTGCGAAATATTTTGAGCGGATTTTCATGTGCATCAATTTAAAATTGATATTTATTTATTTTGTGCTGTCTGGGCTTTTCAATTTCTTCAGAGAAATCCTGGGATCCTCTTCTTTCACCCTCGGATCTCATCCTCTGGACCTTCCATCTCTCTGGTTTCTTCCTCTGTAATTCAACTTATTATATGTACACACCGTCCTCTAAGATTTTGTGGATTCTTTACTTTGCAGGATCTGAAGGAGATTAAAAGGGGGTTAGACCATGGCAAGAGATCAGCTTCAGGTCCTCACCGCGCTAGATGTCGCCAAGACGCAGTGGTATCACTTCACGGCGATCGTGATCGCCGGCATGGGGTTCTTTACCGACGCCTACGACC are provided.

Specifically, the method for obtaining the SEQ ID NO.2 plant promoter in the PCR mode comprises the following steps: SpPHT1 promoter 2 (SpPHT1 gene promoter sequence, SEQ ID NO.2) is amplified by taking total duckweed genome DNA as a template. The PCR reaction system is as follows: DNA 2. mu.L, 10 XPCR Buffer 2.5. mu.L, SpPHT1-Q2-F (SEQ ID No.6) 1. mu.L, dNTP (10mM each) 1. mu.L, SpPHT1-Q1-R (SEQ ID No.5) 1. mu.L, LA Taq 0.5. mu.L, ddH₂O17. mu.L. PCR amplification conditions: pre-denaturation at 95 ℃ for 3 min; denaturation at 95 ℃ for 30s, annealing at 63.1 ℃ for 30s, extension at 72 ℃ for 1min, and 30 cycles; extension at 72 ℃ for 5 min. Wherein: the nucleotide sequence of SpPHT-Q2-F (SEQ ID No.6) is as follows: 5'-CCCAAGCTTTTGTGGTCGTGCGTGTGA-3' (with Hind III cleavage sites at the 5 ' end); the nucleotide sequence of SpPHT1-Q1-R (SEQ ID No.5) is as follows:5'-GCTCTAGAGGTCGTAGGCGTCGGTAA-3' (with an XbaI cleavage site at the 5 ' end).

Preferably, it has any one of the nucleotide sequences shown by (VII), (VIII) or (IX):

(VII) shown as SEQ ID NO. 3;

(VIII) a sequence having at least 50% homology to the sequence shown as SEQ ID No. 3;

(IX) a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more nucleotides in the sequence shown in SEQ ID NO. 3.

The plant promoter of the present application can be obtained by artificial synthesis or by PCR. Specifically, the sequence of the third plant promoter of the present application is shown in SEQ ID No.3 and is named as SpPHT1 promoter 1, the 5' end of SpPHT1 promoter 1 has a plurality of action elements more than SpPHT1 promoter 2, and the nucleotide sequence of SpPHT1 promoter 1 is: ACTGGTAGTGCGGTATCCAGCAAGACAAGTGACAGCCAATTGGTGGCCTATATTAATCGTCGTTAGATCCAGAAGATCACTCCGAGGCGACTTCCGTCACGGCGGTGAGGACTTGCCGAGGAGTTTTGCCGAATTTCATACCGTCGTATCAAGCGGAAGCAGGGGTGGACGGAAGTTGAATTAGGAAGTTCGGCTGGGTCCCCGTCCTCACAAGTGTTGACTTTCAAGTTCAACACAGCGGATATTCCTTCATTTGATACCCATCCGAGGCCCACCACCTTGCCCCTCTCCCTCCCATAAGAATTCCTGGTTCTCCTCTTCCATGCAGCCAGTTCTCTCCCTTCCATTCCATTATCCTCTCCCTCCCTCCCTCCCACCTTCCCATCCTCTCTCTCTCTCTCTCTCTCTGTTCCCTCCCTTCTTCCTGTTTGGCTGGACGTTCTTCGCGGTTGGAGAAGGTATGCTTCCGGAGTGAGTTGTGGATAATTCTCTCTCTCTCTCTCTCTCGCTCTCGCTCTGTTATCGGCCTGACTGTAGAAAGATCATGCGTTTGACGACTTGAACGCGGTCAAGAGAGGGTAGTTCAACAAGTTTGTGGTCGTGCGTGTGACGCTATCTAGTTAATTTTTTCAATCCATGCCGGATCATTTTTTTTTTCTAGCATTAGATAGCGGATTGGGGCACTCGTTGCTCCATCTCGGCTAATTTAACGAAATAATTGGAATGAGAGAGATGCTAAAAAAGGGGTCTTCCGTTTGGATAGTGATGCATCCCTCTGTCTCTCTCTCCTCCGGTAGGGCCTTCAAGGTTTTCGTTGAAACTCATCGGAAGATCAGTCTAAAGTTAAGGCAAGTGTCGTGTCGTCTTTCTCTCTCACGTTCCTGGACCCGTGATCGTTTGATTCCAGGAACTTTGGTGATCGATTGATTCTTGAGAGTCCAAAAATCGGTCAACGACTGCGAAATATTTTGAGCGGATTTTCATGTGCATCAATTTAAAATTGATATTTATTTATTTTGTGCTGTCTGGGCTTTTCAATTTCTTCAGAGAAATCCTGGGATCCTCTTCTTTCACCCTCGGATCTCATCCTCTGGACCTTCCATCTCTCTGGTTTCTTCCTCTGTAATTCAACTTATTATATGTACACACCGTCCTCTAAGATTTTGTGGATTCTTTACTTTGCAGGATCTGAAGGAGATTAAAAGGGGGTTAGACCATGGCAAGAGATCAGCTTCAGGTCCTCACCGCGCTAGATGTCGCCAAGACGCAGTGGTATCACTTCACGGCGATCGTGATCGCCGGCATGGGGTTCTTTACCGACGCCTACGACC are provided.

Specifically, the method for obtaining the SEQ ID NO.3 plant promoter in the PCR mode comprises the following steps: SpPHT1 promoter 1 (SpPHT1 gene promoter sequence, SEQ ID NO.3) is amplified by taking total duckweed genome DNA as a template. The PCR reaction system is as follows: DNA 2. mu.L, 10 XPCR Buffer 2.5. mu.L, SpPHT1-Q3-F (SEQ ID No.7) 1. mu.L, dNTP (10mM each) 1. mu.L, SpPHT1-Q1-R (SEQ ID No.5) 1. mu.L, LA Taq 0.5. mu.L, ddH₂O17. mu.L. PCR amplification conditions: pre-denaturation at 95 ℃ for 3 min; denaturation at 95 ℃ for 30s, annealing at 63.1 ℃ for 30s, extension at 72 ℃ for 1min, and 30 cycles; extension at 72 ℃ for 5 min. Wherein: the nucleotide sequence of SpPHT-Q3-F (SEQ ID No.7) is as follows: 5'-CCCAAGCTTACTGGTAGTGCGGTATCC-3' (with Hind III cleavage sites at the 5 ' end); the nucleotide sequence of SpPHT1-Q1-R (SEQ ID No.5) is as follows: 5'-GCTCTAGAGGTCGTAGGCGTCGGTAA-3' (with an XbaI cleavage site at the 5 ' end).

Specifically, the plant promoters of SEQ ID NO.1 and SEQ ID NO.2 start the specific expression of target genes in the root tissues of plants; the plant promoter of SEQ ID NO.3 can promote the specific expression of target gene in the root and stem tissue of plant.

Preferably, the substitution is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65 nucleotides.

The second aspect of the application discloses the application of the plant promoter in promoting the high-efficiency expression of downstream target genes in plants.

Preferably, the plant is selected from the group consisting of duckweed, tobacco, and Arabidopsis.

Preferably, the expression of the plant promoter in promoting a target gene in a plant is specifically as follows: the plant promoter is used for promoting the specific expression of a target gene in the root, stem or leaf tissue of a plant.

In a third aspect of the present application, an expression vector comprising said plant promoter is disclosed. In particular, the expression vector may be an existing conventional expression vector, such as a recombinant T plasmid plant expression vector.

In a fourth aspect of the present application, a host cell comprising the expression vector is disclosed.

Specifically, the target gene initiated by the present application can be a gene favorable for plant growth, such as a phosphorus transporter gene, which is a transporter having affinity for phosphorus (Pi), and regulates the absorption and transport of phosphorus by plants, and has a relevant influence on the utilization rate of plant phosphorus. Phosphorus is one of the macronutrients essential for plant growth and development, and accounts for about 0.2% of the dry weight of the plant. It plays various basic biological functions as a structural element in nucleic acid and phospholipid, energy metabolism, activation of metabolic intermediates, signal transduction cascade and regulation of enzymes, overexpresses genes in plants which are beneficial to plant growth, and can improve the yield of the plants.

The fifth aspect of the application discloses a method for purifying eutrophic water, which is to plant the plant for purifying the eutrophic water, which contains the host cell, in the eutrophic water, wherein the host cell comprises the plant promoter and the target gene for promoting the growth of the plant. Specifically, the plant promoter promotes the eutrophic water purification plants to grow in a large amount in the eutrophic water, so that the eutrophic water can be effectively purified, the yield of the plants for purifying the eutrophic water can be increased, and the economic benefit of farmers can be guaranteed.

The plant promoter disclosed by the application is derived from duckweed, the sequence of the plant promoter discovered by the application is not reported in any prior art, and the public database such as GenBank and the like does not have any comment on the promoter and the characteristics of the promoter. The invention discovers and identifies the function and the starting efficiency of the promoter for the first time. The method comprises the steps of cloning for the first time to obtain a duckweed SpPHT1 promoter (SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3), obtaining a transgenic plant by utilizing a plant transgenic engineering technical means, and determining the position and the enzymic property of GUS expression of a reporter gene by histochemical staining and a fluorescence spectrophotometry method, so that the expression regulation and control characteristics of the duckweed SpPHT1 promoter are analyzed. The application discovers that a plurality of functional elements in the azolla SpPHT1 promoter sequence are closely related to the control of downstream target gene expression, and the result can provide a new gene resource for purifying eutrophic water and breeding crops.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.

FIG. 1 shows the sequence analysis of SpPHT1 promoter 1 in the example of the present application; wherein AAGAA-motif: an abscisic acid-responsive element; ABRE: an abscisic acid-responsive element; ARE: an induced anaerobic response element; box 4: a light responsive element; CAAT-box: promoter and enhancer regions common elements of action; CARE: a gibberellin-responsive element; CCGTCC-box: regulating meristem-specific active elements; CGTCA-motif: a jasmonic acid responsive element; ERE: an ethylene responsive element; G-Box: a light responsive element; g-box: a photoreaction modulating element; MYB: drought, high salinity, low temperature response elements; MYC: an anti-abscisic acid-responsive element; TATA-box: a transcription core promoter element; TCCC-motif: a light responsive element; TGACG-motif: a jasmonic acid responsive element; GGTGC functional region (denoted Unnamed 1 in the figure): from maize, the function of which is unknown; TSSP: a transcription initiation point;

FIG. 2 is an electrophoresis diagram of a plant promoter (a is SpPHT1 promoter 3; b is SpPHT1 promoter 2; and c is SpPHT1 promoter 1) provided in the examples of the present application;

FIG. 3 shows the staining results of transgenic tobacco and wild-type roots according to the examples of the present application. a) The method comprises the following steps Root of transgenic tobacco Z-SpPHT1-Q1, b): root of transgenic tobacco Z-SpPHT1-Q2, c): root of transgenic tobacco Z-SpPHT1-Q3, d): root, e) of transgenic tobacco Z-SpPHT1-PBI 121: roots of wild-type WT tobacco;

FIG. 4 shows the staining results of transgenic tobacco and wild-type stem according to the example of the present application. a) The method comprises the following steps Stems of transgenic tobacco Z-SpPHT1-Q1, b): stem, c) of transgenic tobacco Z-SpPHT 1-Q2: stem, d) of transgenic tobacco Z-SpPHT 1-Q3: stem, e) of transgenic tobacco Z-SpPHT1-PBI 121: stems of wild-type WT tobacco;

FIG. 5 shows the staining results of transgenic tobacco and wild-type leaves according to the examples of the present application. a) The method comprises the following steps Leaf of transgenic tobacco Z-SpPHT1-Q1, b): leaf of transgenic tobacco Z-SpPHT1-Q2, c): leaf of transgenic tobacco Z-SpPHT1-Q3, d): leaf of transgenic tobacco Z-SpPHT1-PBI121, e): leaves of wild-type WT tobacco;

FIG. 6 is a BSA standard curve provided in examples herein;

FIG. 7 is a MU standard curve provided in accordance with an embodiment of the present application;

FIG. 8 shows the GUS enzyme activity assay of transgenic tobacco Z-SpPHT1-PBI121, transgenic tobacco Z-SpPHT1-Q1, Z-SpPHT1-Q2, Z-SpPHT1-Q3 and wild type WT tobacco in three different tissues of root, stem and leaf.

Detailed Description

The present application provides a novel plant promoter capable of specifically promoting the efficient expression of a gene of interest in a tissue of a rhizome leaf in a plant.

The technical solutions in the embodiments of the present application will be described clearly and completely below, and it should be understood that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

The raw materials and reagents used in the following examples are commercially available or self-made.