Phosphatase coding gene EcPAP19 and application thereof

Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a phosphatase coding gene EcPAP19 and application thereof. The nucleotide sequence of the phosphatase coding gene EcPAP19 is as shown in SEQ ID NO: 1. In the embodiment of the invention, the acid phosphatase gene EcPAP19 is cloned from eichhornia crassipes, and through overexpression of the EcPAP19 gene in rice, it is found that under the condition of organophosphorus, contrast rice is poor in utilization of organophosphorus, so that phosphorus-lacking growth limitation is caused; and the transgenic rice can utilize organic phosphorus, and the growth vigor of the transgenic rice is not obviously changed compared with that of a normal culture medium, so that the EcPAP19 gene is a key gene for promoting the absorption of the plant to the phosphorus element. The invention has important application value in the aspects of enhancing phosphorus absorption of plants and breeding new varieties.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant genetic engineering, and in particular relates to a phosphatase encoding gene EcPAP19 and an application thereof. Background Art

[0002] Phosphorus is an essential nutrient for all living things on Earth. In plants, it is a key element involved in photosynthesis and respiration, and is also a major element in the synthesis of nucleic acids and plasma membranes. However, plants can only obtain phosphorus through passive absorption of inorganic phosphate from the environment through their roots. As a result, the phosphorus fertilizer utilization rate of crops is generally less than 25%, making it crucial to improve its utilization efficiency.

[0003] Therefore, in-depth exploration of the key genes that regulate plant phosphorus absorption is of great significance for in-depth analysis of the molecular mechanism of plant phosphorus absorption and the cultivation of new plant varieties that efficiently utilize phosphorus. Summary of the Invention

[0004] The present invention aims to provide a phosphatase encoding gene EcPAP19 and its application. By overexpressing the EcPAP19 gene in rice, it was found that the transgenic rice can utilize organic phosphorus, indicating that the EcPAP19 gene is a key gene that promotes plant phosphorus absorption.

[0005] The present invention provides a phosphatase encoding gene EcPAP19, the nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO: 1.

[0006] The present invention also provides the use of the phosphatase encoding gene EcPAP19 described above in enhancing phosphorus absorption by plants and / or creating new germplasm with enhanced phosphorus absorption.

[0007] The present invention also provides a primer pair for amplifying the phosphatase encoding gene EcPAP19 described above, wherein the primer pair comprises a forward primer and a reverse primer;

[0008] The nucleotide sequence of the forward primer is shown in SEQ ID NO: 2;

[0009] The nucleotide sequence of the reverse primer is shown in SEQ ID NO: 3.

[0010] The present invention also provides a biological material that overexpresses the phosphatase encoding gene EcPAP19 described above, wherein the biological material comprises one or more of a recombinant expression vector containing the phosphatase encoding gene EcPAP19, a recombinant microorganism containing the phosphatase encoding gene EcPAP19, a transgenic cell line containing the phosphatase encoding gene EcPAP19, a recombinant microorganism containing the recombinant expression vector, and a transgenic cell line containing the recombinant expression vector.

[0011] As a preferred embodiment, the basic skeleton of the recombinant expression vector containing the phosphatase encoding gene EcPAP19 includes the pBWA(V)HS vector.

[0012] The present invention also provides the use of the primer pair or the biological material described above in enhancing the absorption of phosphorus by plants.

[0013] The present invention also provides the use of the above-mentioned primer pair or the above-mentioned biological material in creating a new germplasm with enhanced phosphorus absorption.

[0014] The present invention also provides a method for enhancing plant absorption of phosphorus, comprising the following steps: overexpressing the phosphatase encoding gene EcPAP19 in the target plant genome to obtain a plant with enhanced phosphorus absorption; the nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO: 1.

[0015] The present invention also provides a method for creating a new germplasm with enhanced phosphorus absorption, characterized in that it includes the following steps: overexpressing the phosphatase encoding gene EcPAP19 in the target plant genome to obtain the new germplasm with enhanced phosphorus absorption; the nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO: 1.

[0016] As a preferred embodiment, the plant includes rice.

[0017] Beneficial effects: The present invention provides a phosphatase encoding gene EcPAP19, the nucleotide sequence of which is shown in SEQ ID NO: 1. In one embodiment of the present invention, the acid phosphatase gene EcPAP19 was cloned from water hyacinth. By overexpressing the EcPAP19 gene in rice, it was found that under organophosphate conditions, the control rice had poor organic phosphorus utilization, resulting in phosphorus deficiency and growth restriction; while the transgenic rice could utilize organic phosphorus, and its growth was not significantly different from that in normal culture medium, indicating that the EcPAP19 gene is a key gene that promotes plant phosphorus absorption. The present invention has important application value in enhancing plant phosphorus absorption and breeding new varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below.

[0019] Figure 1 is the phylogenetic tree of EcPAP19 protein;

[0020] Figure 2 for the subcellular localization of EcPAP19 protein;

[0021] Figure 3 The utilization of organic and inorganic phosphorus by transgenic rice and wild-type rice. DETAILED DESCRIPTION

[0022]

[0023] As a specific embodiment, the phosphatase encoding gene EcPAP19 was cloned from a water hyacinth plant and is the purple acid phosphatase encoding gene EcPAP19 of water hyacinth. As a specific embodiment, acid phosphatases are a family of hydrolases commonly found in organisms that can hydrolyze organophosphorus compounds under acidic conditions. When plants are exposed to low inorganic phosphate, their roots can secrete acid phosphatase to degrade organophosphorus compounds into inorganic phosphate for plant absorption. Purple acid phosphatase is an important component of acid phosphatase, named for the purple color of its aqueous solution. It has a relatively wide pH adaptability and temperature stability and can act on a wide range of substrates. Under weakly acidic conditions of pH 4.0 to 7.0, purple acid phosphatase can catalyze the hydrolysis of organic phosphate monoesters and anhydrides and release inorganic phosphate that can be absorbed by plants, thereby improving phosphorus utilization efficiency in soil or water. It plays a key role in plant phosphorus absorption and utilization.

[0024] The present invention also provides the use of the aforementioned phosphatase-encoding gene, EcPAP19, for enhancing plant phosphorus uptake and / or creating new germplasm with enhanced phosphorus uptake. As a specific embodiment, the present invention cloned an acid phosphatase, EcPAP19, from the plant Eichhornia crassipes. Transcriptome and fluorescence quantitative expression analysis demonstrated that EcPAP19 plays an important role in phosphorus uptake in Eichhornia crassipes.

[0025] The present invention also provides a primer pair for amplifying the phosphatase encoding gene EcPAP19 described above, wherein the primer pair comprises a forward primer and a reverse primer;

[0026] The nucleotide sequence of the forward primer is shown in SEQ ID NO: 2;

[0027] The nucleotide sequence of the reverse primer is shown in SEQ ID NO: 3.

[0028] The sequences of the primer pairs of the present invention are as follows:

[0029] Forward primer (SEQ ID NO: 2): 5′-ATGGCCGGGAGGTCCTCAGG-3′;

[0030] Reverse primer (SEQ ID NO: 3): 5'-TACAGAGCTGGGTTCAGGTGAAG-3'.

[0031] The present invention also provides a biomaterial that overexpresses the phosphatase-encoding gene EcPAP19 described above. The biomaterial comprises one or more of a recombinant expression vector containing the phosphatase-encoding gene EcPAP19, a recombinant microorganism containing the phosphatase-encoding gene EcPAP19, a transgenic cell line containing the phosphatase-encoding gene EcPAP19, a recombinant microorganism containing the recombinant expression vector, or a transgenic cell line containing the recombinant expression vector. The basic framework of the recombinant expression vector containing the phosphatase-encoding gene EcPAP19 of the present invention comprises a pBWA(V)HS vector. In one embodiment, the recombinant expression vector containing the phosphatase-encoding gene EcPAP19 comprises a pBWA(V)HS-EcPAP19 overexpression vector obtained by inserting the EcPAP19 gene into a pBWA(V)HS vector. In one embodiment, the recombinant microorganism containing the phosphatase-encoding gene EcPAP19 comprises Agrobacterium tumefaciens GV3101 containing the EcPAP19 gene.

[0032] The present invention also provides the use of the primer pair or the biological material described above in enhancing the absorption of phosphorus by plants.

[0033] The present invention also provides the use of the above-mentioned primer pair or the above-mentioned biological material in creating a new germplasm with enhanced phosphorus absorption.

[0034] The present invention also provides a method for enhancing phosphorus absorption in plants, comprising the following steps: overexpressing the phosphatase encoding gene EcPAP19 in the genome of a target plant to obtain a plant with enhanced phosphorus absorption; the nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO: 1. In an embodiment of the present invention, the pBWA(V)HS-EcPAP19 overexpression vector is introduced into Agrobacterium GV3101, and the overexpression vector is transformed into the genome of the target plant using an Agrobacterium-mediated transgenic method to obtain a plant with enhanced phosphorus absorption.

[0035] The present invention also provides a method for creating a new germplasm with enhanced phosphorus absorption, comprising the steps of overexpressing the phosphatase-encoding gene EcPAP19 in the genome of a target plant to obtain the new germplasm with enhanced phosphorus absorption; the nucleotide sequence of the phosphatase-encoding gene EcPAP19 is shown in SEQ ID NO: 1. In an embodiment of the present invention, the pBWA(V)HS-EcPAP19 overexpression vector is introduced into Agrobacterium GV3101, and the overexpression vector is transformed into the genome of the target plant using Agrobacterium-mediated transgenic methods to obtain the new germplasm with enhanced phosphorus absorption. The plants described in the present invention include rice.

[0036] Specific examples of the present invention show that after overexpressing the phosphatase encoding gene EcPAP19 in Nipponbare rice, under organophosphate conditions, the control rice has poor utilization of organophosphate, resulting in limited growth due to phosphorus deficiency; while the transgenic rice can utilize organophosphate, and its growth does not change significantly from that in normal culture medium. This indicates that the transgenic rice can secrete EcPAP19 protein and deliver it outside the root system, hydrolyzing organophosphate in the water into inorganic phosphate, thereby promoting the plant's absorption of phosphorus.

[0037] To further illustrate the present invention, a phosphatase encoding gene EcPAP19 and its application provided by the present invention are described in detail below in conjunction with examples, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Unless otherwise specified, the present invention has no special requirements for the raw materials, and commercially available products known to those skilled in the art can be used.

[0039] Example

[0040] 1. Extraction of Total RNA and cDNA Synthesis from Eichhornia crassipes: Eichhornia crassipes samples were quickly frozen in liquid nitrogen and ground into powder. Total RNA was extracted using the RNAsimple Total RNA Kit (Beijing Tiangen Biochemical Technology Co., Ltd.). Total RNA was reverse transcribed into cDNA using the HiScript III 1st Strand cDNA Synthesis Kit (Nanjing Novozymes Biotech Co., Ltd.).

[0041] 2. Cloning of the Eichhornia crenata purple acid phosphatase gene EcPAP19: The Eichhornia crenata EcPAP19 gene sequence is shown in SEQ ID NO: 1. Based on this sequence, a pair of primers were designed: forward primer: 5'-ATGGCCGG GAGGTCCTCAGG-3' (SEQ ID NO: 2) and reverse primer: 5'-TACAGAGCTGGGTTC AGGTGAAG-3' (SEQ ID NO: 3). PCR amplification was performed using the cDNA reverse-transcribed in step 1 as a template. The PCR reaction conditions were: 94°C pre-denaturation for 30 seconds; 98°C denaturation for 10 seconds, 55°C annealing for 5 seconds, and 72°C extension for 30 seconds for a total of 30 cycles; and 72°C extension for 10 minutes. The PCR products were electrophoresed on a 1% agarose gel, and bands of the appropriate length were cut out using a gel imaging system. The fragments were recovered using the FastPure Gel DNA Extraction Mini Kit-DC301 (Nanjing Novozymes Biotechnology Co., Ltd.), and then amplified by PCR using primers with homology arms (CS554-EcPAP19-F: 5'-catttggagaggacacgcATGGCCGG GAGGTCCTCAGG-3' (SEQ ID NO: 4); CS554-EcPAP19-R: 5'-tcgcccttgctcac catgaaTACAGAGCTGGGTTCAGGTGAAG-3' (SEQ ID NO: 5), where lowercase letters represent homology arms). After length detection by electrophoresis, the fragments were excised from the gel and recovered using the same steps as above. The recovered fragments were stored at -20°C.

[0042] 3. Sequence alignment and phylogenetic tree construction: All protein sequences of PAP gene family members in Eichhornia crassipes, Pseudomonas aeruginosa, Arabidopsis thaliana, and rice were aligned using MAFFT, and the aligned sequence files were then used to construct a phylogenetic tree using IQ-TREE. The parameters were modified so that the tree construction method was based on the most appropriate model after calculation, and the bootstrap value was 1000.

[0043] 4. Real-time quantitative PCR reaction: Total RNA from different parts of Eichhornia crassipes (stolons, roots, and leaves) treated with different phosphorus concentrations (DP: 0 mg / L, LP: 5.0 mg / L, MP: 50 mg / L, HP: 100 mg / L) was used as a template (RNA extraction was performed according to step 1). Forward primer: 5'-AGCACAGATTCCATACGCCTTAC-3' (SEQ ID NO: 6) and reverse primer: 5'-CGTCTCGCTTCGGTTCACTT-3' (SEQ ID NO: 7) were used as primers. Real-time quantitative PCR reaction was performed using ChamQ SYBR qPCR Master Mix (Nanjing Novozymes Biotech Co., Ltd.) with EcUBC as the internal reference gene. The extracted total RNA was treated with the following system to remove genomic DNA: RNase-free ddH2O 16 μL, 4×gDNAwiperMix 4 μL, RNA 1 pg -1 The reverse transcription reaction system consisted of 4 μL of 5× HiScript III qRT SuperMix and 16 μL of the first step reaction solution. The reaction temperature was 37°C for 15 min and 85°C for 5 s.

[0044] 5. Subcellular localization: pART-CAM-EGFP-CS544 (Nanjing Jisi Huiyuan Biotechnology Co., Ltd.) was double-digested with EcoRI and XhoI (Thermo Fisher Scientific). The digestion reaction consisted of 1 μg of plasmid, 2 μL of 10× FastDigest Green Buffer, 1 μL of XhoI, 1 μL of EcoRI, and 20 μL of ddH2O. The mixture was placed in a PCR instrument and digested at 37°C for 15 min. The digested product was electrophoresed on a 1% agarose gel and recovered using a gel imaging system (same as step 2). The pART-CAM-EGFP-CS544-EcPAP19 recombinant plasmid was constructed by homologous recombination using 300 ng of linearized vector, 60 ng of insert, 5 μL of 2×CE Mix, and 10 μL of ddH2O. The plasmid was cloned using DH5α and transformed into GV3101 Agrobacterium (Nanjing Enzymepulis Biotechnology Co., Ltd.) with the following steps: 1) Remove the GV3101 competent cell from -80°C and quickly place it on ice to thaw for 5 minutes. 2) Add the pART-CAM-EGFP-CS544-EcPAP19 recombinant plasmid and mix well. Place it on an ice box for 5 minutes, in liquid nitrogen for 5 minutes, in a 37°C water bath for 5 minutes, and in an ice bath for 5 minutes. 3) Add 900 μL of YEB liquid medium without antibiotics, mix well, and culture at 28°C with a shaker at 200 rpm for 3 hours. 4) Centrifuge at 6000 rpm for 1 minute, discard 900 μL of supernatant, retain 100 μL of supernatant, gently pipette the resuspended bacterial block, and spread it on a YEB plate containing 1‰ Kan. Place it upside down in a 28°C incubator for 3 days. The transformed bacterial solution was evenly spread on YEB solid medium containing 1‰ Rif and 1‰ Kan, and cultured in a 28℃ incubator for 60h. A sterilized white pipette tip was used to pick up a single colony, placed in a 2mL sterilized centrifuge tube, and 1mL of YEB liquid medium containing 1‰ Rif and 1‰ Kan was added. The culture was incubated at 200rpm and 28℃ until the bacterial solution became turbid. 100μL of bacterial solution was taken into a sterilized 150mL conical flask, and 50mL of YEB liquid medium containing 1‰ Rif and 1‰ Kan was added. The culture was incubated at 28℃ and 200rpm until the OD 600 The bacterial solution was transferred to a 50 mL sterile centrifuge tube, centrifuged at 3000 rpm for 10 min, and the supernatant was discarded. The suspension was resuspended with 10 mM MgCl2, 10 mM MES and 100 μM acetosyringone to an OD of 0. 600 The suspension was placed in the dark for 3 hours and then injected into the tobacco leaves from the back with a sterile syringe. After culturing overnight for 12 hours, the leaves were cultured under normal light conditions for 48 hours and observed under a confocal microscope.

[0045] 6. Transgenic verification: transgenic vector construction and rice transformation: The EcPAP19 gene was amplified using specific primers (forward primer: 5'-ATGGCCGGGAGGTCCTCAGG-3' (SEQ ID NO: 2), reverse primer: 5'-TACAGAGCTGGGTTCAGGTGAAG-3' (SEQ ID NO: 3)) and then constructed into the pBWA(V)HS vector (Li Q, Xu J, Yang L, Zhou X, Cai Y and Zhang Y (2020) Transcriptome Analysis of Different Tissues Reveals Key Genes Associated With Galanthamine Biosynthesis in Lycoris longituba. Front. Plant Sci. 11: 519752. doi: 10.3389 / fpls.2020.519752) to form the 35S overexpression vector pBWA(V)HS-EcPAP19. The recombinant vector pBWA(V)HS-EcPAP19 was transformed into Agrobacterium tumefaciens strain GV3101 by electroporation (same method as in step 5), and then transformed into the rice variety Nipponbare using a callus method (Hiei, Y., Ohta, S., Komari, T., and Kumashiro, T. (1994). Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6:271-282.). Nipponbare containing an empty pBWA(V)HS-gbd vector was used as a control. The control and transgenic plants were cultured in Yoshida medium (Coolaber) for complete rice and Yoshida medium containing organic phosphorus instead of inorganic phosphorus. The rice growth was observed after 7 days of culture.

[0046] 7. Experimental Results

[0047] 1) The protein sequence encoded by the cloned EcPAP19 gene was compared with other PAPs in water hyacinth, rice PAP, PAP of pickerelleaf, and PAP of Arabidopsis thaliana to construct a phylogenetic tree. The results showed that EcPAP19 had the closest evolutionary relationship with PcPAP24 of pickerelleaf and OsPAP10 of rice ( Figure 1 ).

[0048] 2) Under phosphorus deficiency conditions, EcPAP19 was abundantly expressed in the roots of Eichhornia crassipes, indicating that it plays an important role in the phosphorus uptake of the roots (Table 1).

[0049] 3) Subcellular localization EcPAP19 protein is located outside the cell, indicating that EcPAP19 protein can be secreted outside the root system ( Figure 2 ).

[0050] 4) EcPAP19 can be expressed in the rice variety Nipponbare (Table 2). Under organic phosphorus conditions, the control rice poorly utilizes organic phosphorus, resulting in phosphorus-deficient growth restriction; however, the transgenic rice can utilize organic phosphorus, and its growth is not significantly different from that in normal culture medium (Tables 3 and Figure 3 ), indicating that transgenic rice can secrete EcPAP19 protein and deliver it to the outside of the root system, hydrolyzing organic phosphorus in the water into inorganic phosphate, and promoting the plant's absorption of phosphorus.

[0051] Table 1 Analysis of the expression of EcPAP19 gene in different tissues of Eichhornia crassipes

[0052] organize Phosphorus treatment <![CDATA[Relative expression level (2 -ΔΔCt )]]> Standard deviation Significance of difference (same tissue) runners DP 3.245 1.214 a runners LP 0.552 0.217 b runners MP 0.373 0.156 b runners HP 0.410 0.260 b root DP 16.955 6.507 a root LP 2.604 0.563 b root MP 3.249 1.387 b root HP 0.945 0.607 b leaf DP 4.272 2.081 a leaf LP 0.364 0.087 b leaf MP 0.141 0.217 b leaf HP 0.136 0.130 b

[0053] Table 2 Expression of EcPAP19 gene in transgenic rice

[0054] Sample name <![CDATA[Relative expression level (2 -ΔΔCt )]]> Standard deviation CK Complete Nutrition 1 0.000 0.000 CK Complete Nutrition 2 0.000 0.000 CK Complete Nutrition 3 0.000 0.000 CK Organophosphorus 1 0.000 0.000 CK Organophosphorus 2 0.000 0.000 CK Organophosphorus 3 0.000 0.000 GM Complete Nutrition 1 9316.174 962.421 GM Complete Nutrition 2 34606.265 3054.709 GM Complete Nutrition 3 76261.748 1977.358 Genetically modified organophosphate 1 39497.078 4569.282 Genetically modified organophosphate 2 45710.420 6823.594 Genetically modified organophosphate 3 5537.109 420.801

[0055] Table 3 Utilization of organic and inorganic phosphorus by transgenic rice and wild-type rice

[0056] deal with Plant height Standard deviation Significance CK organophosphate 24.7 1.99833 b CK complete nutrition 29.125 1.98053 a Genetically modified organophosphates 30.6 1.57692 a Genetically modified complete nutrition 29.15 0.91469 a

[0057] Thus, the present invention cloned the acid phosphatase gene EcPAP19 in water hyacinth and overexpressed the EcPAP19 gene in rice. It was found that under organophosphate conditions, the control rice poorly utilized the organic phosphorus, resulting in phosphorus-deficient growth restriction. However, the transgenic rice could utilize the organic phosphorus, and its growth was not significantly different from that in normal culture medium. This indicates that the EcPAP19 gene is a key gene that promotes plant phosphorus absorption. The present invention has important application value in enhancing plant phosphorus absorption and breeding new varieties.

[0058] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A phosphatase encoding gene EcPAP19, characterized in that: The nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO:

1.

2. Use of the phosphatase encoding gene EcPAP19 according to claim 1 in enhancing phosphorus absorption in plants and / or creating new germplasm with enhanced phosphorus absorption.

3. A primer pair for amplifying the phosphatase encoding gene EcPAP19 according to claim 1, characterized in that: The primer pair includes a forward primer and a reverse primer; The nucleotide sequence of the forward primer is shown in SEQ ID NO: 2; The nucleotide sequence of the reverse primer is shown in SEQ ID NO:

3.

4. A biomaterial that overexpresses the phosphatase encoding gene EcPAP19 according to claim 1, characterized in that: The biological material includes one or more of a recombinant expression vector containing the phosphatase encoding gene EcPAP19, a recombinant microorganism containing the phosphatase encoding gene EcPAP19, a transgenic cell line containing the phosphatase encoding gene EcPAP19, a recombinant microorganism containing the recombinant expression vector, and a transgenic cell line containing the recombinant expression vector.

5. The biomaterial according to claim 4, characterized in that The basic skeleton of the recombinant expression vector containing the phosphatase encoding gene EcPAP19 includes the pBWA(V)HS vector.

6. Use of the primer pair according to claim 3 or the biomaterial according to claim 4 or 5 for enhancing phosphorus absorption by plants.

7. Use of the primer pair according to claim 3 or the biomaterial according to claim 4 or 5 in creating a new germplasm with enhanced phosphorus absorption.

8. A method for enhancing phosphorus absorption by plants, characterized in that: The following steps are involved: The phosphatase encoding gene EcPAP19 is overexpressed in the target plant genome to obtain a plant with enhanced phosphorus absorption; the nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO:

1.

9. A method for creating a new germplasm that enhances phosphorus absorption, characterized in that: The following steps are involved: The phosphatase encoding gene EcPAP19 is overexpressed in the target plant genome to obtain the new germplasm with enhanced phosphorus absorption; the nucleotide sequence of the phosphatase encoding gene EcPAP19 is shown in SEQ ID NO:

1.

10. The method according to claim 9, characterized in that The plants include rice.

Images