Sugarcane ScPHR1 gene, expression cassette and application thereof

Abstract

The invention relates to the technical field of gene engineering, in particular to a sugarcane ScPHR1 gene, an expression cassette and application of the sugarcane ScPHR1 gene. The invention provides application of a sugarcane ScPHR1 gene as shown in SEQ ID NO.1 in improvement of low phosphorus stress tolerance and gray mold resistance of plants, which comprises the following steps: constructing a recombinant expression vector containing the sugarcane ScPHR1 gene as shown in SEQ ID NO.1, transforming the recombinant expression vector into agrobacterium tumefaciens, and culturing to obtain recombinant bacteria carrying the recombinant expression vector, and infecting the plant by using the recombinant bacteria carrying the recombinant expression vector. After the sugarcane ScPHR1 gene obtained by the invention is over-expressed in arabidopsis thaliana, the tolerance of the arabidopsis thaliana to low phosphate can be improved, and the disease resistance of the arabidopsis thaliana to botrytis cinerea is improved. And a basis is provided for screening excellent transgenic plants.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, and more particularly to a sugarcane ScPHR 1. Genes, expression cassettes and their applications. Background Technology

[0002] sugar cane( Saccharum Sugarcane (spp.), a pillar of the global sugar industry and an emerging energy crop, belongs to the Poaceae subtribe of the Sorghum tribe. It contributes over 80% of the world's sugar production and 40% of its bioethanol production. With breakthroughs in whole-genome sequencing technology, the research team successfully completed de novo haplotype chromosome-level genome assembly of the hybrid sugarcane variety ZZ1. In sugarcane functional genomics research, MYB transcription factors have attracted significant attention due to their diverse regulatory potential.

[0003] The MYB family is the largest family of transcription factors in plants, initially discovered through the regulation of anthocyanin biosynthesis in the aleurone layer of maize. This family is characterized by a conserved MYB base-binding domain at its N-terminus, typically composed of one or more incomplete tandem repeat sequences. Due to their wide range of roles in numerous biological processes, MYB transcription factors play an indispensable role in regulating plant development, mediating hormone signaling pathways, and enhancing tolerance to abiotic stresses. MYB-type coil-coil (MYB-CC) transcription factors are a special subgroup within the MYB superfamily, characterized by the simultaneous presence of a conserved MYB domain and a predicted coil-coil (CC) domain. These two domains jointly promote protein dimerization and functional regulation. Existing research has clearly demonstrated that MYB-CC transcription factors play a crucial role in mediating plant responses to phosphorus (Pi) deficiency stress and regulating the expression of phosphorus starvation-induced genes. Arabidopsis thaliana (… Arabidopsis thaliana Phosphorus starvation response protein 1 (AtPHR1) is the first MYB-CC protein identified to participate in plant phosphorus stress response. In Arabidopsis, PHR1 acts as a pivotal regulator of the phosphorus starvation signaling pathway, regulating the expression of key genes, including members of the phosphorus transporter PHT1 family, by binding to the PHR1 binding sequence (P1BS, GNATATNC) cis-acting element in the promoter of phosphorus starvation-induced (PSI) genes. Under phosphorus-deficient conditions, Arabidopsis... phr1The mutant exhibits multi-faceted phosphorus response disorders: on the one hand, the distribution pattern of phosphorus among root caps is changed, accompanied by decreased anthocyanin and carbohydrate accumulation and adjusted lipid composition and reshaped gene expression; on the other hand, the expression induction ability of multiple phosphate starvation-induced genes is also significantly impaired. However, overexpression of PHR1 in Arabidopsis can significantly up-regulate the expression of specific phosphate starvation-induced genes, thereby promoting the increase of phosphorus content in the plant body, accompanied by the increase of anthocyanin accumulation. Extensive studies have confirmed that this regulatory mechanism is widely conserved in evolution, and the functional orthologous genes of Arabidopsis AtPHR1 have been identified in Chlamydomonas reinhardtii (CrPSR1), rice (OsPHR2), corn (ZmPHR1) and soybean (GmPHR1) and other photosynthetic organisms; in addition to regulating phosphorus homeostasis, members of this family have also been reported to be involved in plant growth promotion, drought resistance enhancement and metal stress response and other biological processes. So far, the function of MYB-CC type genes in sugarcane has not been clearly analyzed, and its regulatory mechanism and biological significance are still unknown. SUMMARY

[0004] The purpose of the present application is to provide a sugarcane ScPHR 1 gene, an expression cassette and application thereof 。

[0005] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions: The present application provides a sugarcane Saccharum officinarum Saccharum officinarum ScPHR 1 gene in improving plant low-phosphorus stress tolerance and resistance to gray mold, comprising the following steps: constructing a recombinant expression vector containing the sugarcane Saccharum officinarum Saccharum officinarum ScPHR 1 gene, transforming the recombinant expression vector into Agrobacterium, culturing to obtain a recombinant bacterium carrying the recombinant expression vector, and using the recombinant bacterium carrying the recombinant expression vector to infect plant plants; The construction method of the recombinant expression vector is: connecting the sugarcane Saccharum officinarum Saccharum officinarum ScPHR 1 gene and pBI121-GFP vector or pCambia2300 vector; The Agrobacterium is GV3101 or EHA105; The plant is Arabidopsis or sugarcane.

[0006] The present application also provides the application of the sugarcane Saccharum officinarum Saccharum officinarum ScPHR 1 gene in cultivating transgenic plants with low-phosphorus stress capacity and resistance to gray mold; the plant is Arabidopsis or sugarcane.

[0007] The present application also provides a sugarcane Saccharum officinarum Saccharum officinarum ScPHR1. The application of a protein encoded by a gene in improving the low phosphorus stress tolerance and resistance to gray mold of a plant; the protein is shown as SEQ ID NO. 2.

[0008] The application further provides a sugarcane ScPHR 1. The application of a protein encoded by a gene in cultivating a transgenic plant with low phosphorus stress tolerance and resistance to gray mold; the plant is Arabidopsis thaliana or sugarcane.

[0009] The application further provides an expression cassette of a sugarcane ScPHR 1. gene, comprising any of the following: (1) a recombinant expression vector: connecting a sugarcane ScPHR 1. gene with a pBI121-GFP vector or a pCambia2300 vector; (2) a recombinant bacterium: transferring a sugarcane ScPHR 1. gene or a recombinant expression vector into Agrobacterium GV3101 or EHA105.

[0010] The application further provides the application of the expression cassette in improving the low phosphorus stress tolerance and resistance to gray mold of a plant; the plant is Arabidopsis thaliana or sugarcane.

[0011] The application further provides the application of the expression cassette in cultivating a transgenic plant with low phosphorus stress tolerance and resistance to gray mold; the plant is Arabidopsis thaliana or sugarcane.

[0012] The application further provides a reagent for detecting the expression amount of a sugarcane ScPHR 1. gene in a plant body in the evaluation of the low phosphorus stress tolerance of a plant; the reagent is a primer pair; the primer pair is shown as SEQ ID NO. 11-12.

[0013] The application further provides a reagent for detecting the expression amount of a sugarcane ScPHR 1. gene in a plant body in the evaluation of the resistance to gray mold of a plant; the reagent is a primer pair; the primer pair is shown as SEQ ID NO. 11-12.

[0014] The application further provides a method for improving the low phosphorus stress tolerance and resistance to gray mold of a plant, overexpressing a sugarcane ScPHR 1. gene in a plant body; or constructing a recombinant expression vector containing a sugarcane ScPHR 1. gene, transforming Agrobacterium with the recombinant expression vector, culturing to obtain a recombinant bacterium carrying the recombinant expression vector, and using the recombinant bacterium carrying the recombinant expression vector to infect a plant. the Agrobacterium is GV3101 or EHA105; the plant is Arabidopsis thaliana or sugarcane.

[0015] The application has the following advantages: The application clones a sugarcane phosphate starvation response gene ScPHR1 Subcellular localization shows that, ScPHR1 the ScPHR1 protein is mainly located in the nucleus.The application also constructs a 35S promoter-driven ScPHR1 overexpression vector, and obtains transgenic Arabidopsis thaliana, and the transgenic Arabidopsis thaliana shows resistance to low-phosphorus stress and gray mold. The application provides a theoretical basis for cultivating new germplasm resistant to low-phosphorus stress and fungal diseases.

[0016] The application preliminarily discloses a sugarcane phosphate starvation response gene ScPHR1 As a new type of plant stress factor, the ScPHR1 protein has potential application value in improving the resistance of plants to low-phosphorus stress and fungal diseases, enriches the functional cognition of the sugarcane phosphate starvation response protein in plant immunity, provides potential genetic resources for disease prevention and control of sugarcane and its crops, and has good application prospect and popularization value. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 is the primary structure of the ScPHR1 protein; Figure 2 is the subcellular localization of the ScPHR1 protein in tobacco; Figure 3 is a TA cloning plasmid containing ScPHR1 the gene; Figure 4 is the relative expression amount determination result of the wild type and the transgenic Arabidopsis thaliana; ScPHR1 Figure 5 is the seed germination condition statistics of the wild type and the transgenic Arabidopsis thaliana seeds in the normal culture medium and the low-phosphorus culture medium (the upper left graph represents the seed germination condition of the Arabidopsis thaliana seeds grown in the normal culture medium; the upper right represents the seed germination condition of the Arabidopsis thaliana seeds grown in the low-phosphorus culture medium; the lower left graph represents the seed germination rate statistics of the Arabidopsis thaliana seeds grown in the normal culture medium; and the lower right graph represents the seed germination rate statistics of the Arabidopsis thaliana seeds grown in the low-phosphorus culture medium; CK represents the normal culture medium; LP is the low-phosphorus culture medium; and WT represents the wild type); Figure 6 is the leaf area statistics result of the wild type and the transgenic Arabidopsis thaliana seeds in the normal culture medium and the low-phosphorus culture medium (CK represents the normal culture medium; LP is the low-phosphorus culture medium; and WT represents the wild type); Figure 7The growth and the result of the electrolyte leakage assay of wild type and transgenic Arabidopsis thaliana after spraying spores of Botrytis cinerea. DETAILED DESCRIPTION

[0018] Sugarcane ScPHR

[0019] Sugarcane ScPHR 1 Protein encoded by the gene, as shown in SEQ ID NO. 2, SEQ ID NO. 2: MQSQKSRVLGAMSSSLPILPNPLKGSFPKPCNPQHIPMSRQLPDDSMPLRNDIHQSASLHPKAGVIGAPYSGYSASPLDSVSNHDSQLMVAPYISQSSIFEAFQSLSDNTPGTHTEAAWFTSSMDVSPLYTDNIAAPDDNQIQSIRPAITSDETAKQNDWWADIMNDDWKDILDATATDSRSKAMIQTSNSATSLPAVNQSASSHSMEICPVASPPNSSNASVAKQRMRWTPELHECFVDAVNQLGGSEKATPKGVLKLMKVDGLTIYHVKSHLQKYRTARYKPDLSEGTSEKRTATEELVLDLKTSMDLTEALRLQMEVQKRLHEQLEIQRKLQLRIEEQGKYLQMMFEKHSRSSMEKVQDPSSRDTVAKPSTLSQSANKDNCATMDPDRTGDSAKTAELGEGSSGLGVKQKLVEIESDAEGATDDGSKISQEKRRKLQDS.

[0020] Sugarcane of SEQ ID NO. 1 ScPHR 1 Primer pair for detecting the expression amount of the gene in plants, as shown in SEQ ID NO. 3~4; SEQ ID NO. 3 (upstream primer): 5'-ATGCAGTCTCAAAAGAGCAGA-3'; SEQ ID NO. 4 (downstream primer): 5'-TTAACTATCTTGCAGTTTGCGC-3'.

[0021] The technical solutions provided by the present application will be described in detail below in combination with examples, but they should not be understood as limiting the scope of protection of the present application.

[0022] Example 1

[0023] 1, Sugarcane total RNA extraction and RT-qPCR detection

[0024] Total RNA was extracted from the sugarcane variety “Zhongzhe No. 1” using the Eastep® Super Total RNA Extraction Kit (Promega). Using 1 μg of total RNA as a template, reverse transcription was performed using the HiScript II First-Strand cDNA Synthesis Kit (Vazyme). Real-time quantitative PCR was performed on a LightCycler 96 system (Roche) using AceQ® qPCR SYBR® Green Master Mix (Vazyme). The PCR amplification program was set as follows: 95℃ pre-denaturation for 30 s; followed by 40 cycles of amplification, each cycle consisting of 95℃ for 10 s, 60℃ for 30 s, and a final hold at 95℃ for 10 s.

[0025] 2. Sugarcane ScPHR 1. Acquisition of genes

[0026] Using the sugarcane genome database (NCBI accession number: GWHEQVP00000000) as a reference, forward primer ScPHR1-F (5'-ATGCAGTCTCAAAAGAGCAGA-3', SEQ ID NO.3) and reverse primer ScPHR1-R (5'-TTAACTATCTTGCAGTTTGCGC-3', SEQ ID NO.4) were designed. Using the obtained cDNA as a template, PCR amplification was performed using the high-fidelity enzyme PhantaMax Super-Fidelity DNA Polymerase (Vazyme). The reaction system was as follows: 25 μL 2×Phanta Master Mix, 1 μL forward primer (10 μM), 1 μL reverse primer (10 μM), 2 μL cDNA, and dd... O was added to a final volume of 50 μL. The PCR reaction program was set as follows: 95℃ pre-denaturation for 30 s; followed by 32 cycles: 95℃ denaturation for 15 s, 60℃ annealing for 15 s, 72℃ extension for 30 s; and a final extension at 72℃ for 10 min after the cycle.

[0027] The PCR product was ligated into a TA cloning vector, transformed into *E. coli* DH5α competent cells, and positive clones were screened and sequenced for verification. Sequencing confirmed the formation of [the desired clone]. ScPHR1The open reading frame (ORF) of the gene, with a nucleotide sequence shown in SEQ ID NO.1, is 1329 bp in length. BlastX alignment analysis (http: / / www.ncbi.nlm.nih.gov) indicates that this ORF encodes 442 amino acids, and the predicted amino acid sequence of the ScPHR1 protein is shown in SEQ ID NO.2. This protein lacks an N-terminal signal peptide and transmembrane domains. Conserved domains were predicted using the Conserved Domains database (https: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi).

[0028] The results show that ScPHR1 has two structural domains, MYB and CC, and belongs to the MYB-CC family. Figure 1 ).

[0029] Example 2

[0030] ScPHR1 Subcellular Localization in Tobacco

[0031] Will ScPHR1 The full-length coding sequence (CDS) was cloned into the pBI121 vector fused with green fluorescent protein (GFP) (pBI121-GFP) to obtain the recombinant expression vector. The recombinant expression vector was then transformed into Agrobacterium tumefaciens strain GV3101. Positive transformants were picked and cultured in LB medium containing 50 mg / L Kan and 20 mg / L Rif (rifampicin) at 28°C with shaking for 2 days. The bacterial cells were collected by centrifugation at 4000 rpm for 5 min, and the bacterial concentration was adjusted to OD600 of 0.7 using Agrobacterium resuspension (10 mmol / L MgCl2, 10 mmol / L 2-(N-morpholino)ethanesulfonic acid, 150 μmol / L acetylsyleugenone). Select tobacco leaves that are 5-6 leaf-aged, uniformly grown, and in good condition. Using a sterile syringe, inject a resuspended solution containing ScPHR1 into the underside of the tobacco leaves. Return the tobacco to the plant culture incubator for further cultivation. After 24 hours of darkness followed by 24 hours of light exposure, observe and capture the fluorescence signals of GFP and red fluorescent protein (RFP) using an Olympus FV3000 laser confocal microscope. Results are as follows: Figure 2 The data shows that ScPHR1 is primarily located in the cell nucleus. This indicates that the gene mainly functions within the cell nucleus.

[0032] Example 3

[0033] Arabidopsis transformation experiment

[0034] 1. Construction of overexpression vectors

[0035] A ScPHR1 hyperrecombinant expression vector driven by the 35S promoter was constructed. It contains... ScPHR1 TA cloning plasmid of gene ( Figure 3 Using a template, specific primers were designed: the forward primer ScPHR1-2300-F (5'-CAGGTCGACTCTAGAGGATCCATGCAGTCTCAAAAGAGCAGA-3', SEQ ID NO.5); and the reverse primer ScPHR1-2300-R (5'-TGGGTACATACGCGTGGTACCTTAACTATCTTGCAGTTTGCGC-3', SEQ ID NO.6). The PCR amplification reaction system and procedure were the same as in Example 2. Subsequently, following the seamless cloning method of Example 2, the purified amplification product was ligated into the pCambia2300 vector, which had been linearized by double digestion with SacⅠ and XbaⅠ. The ligation product was transformed into E. coli DH5α competent cells, and positive clones were screened by colony PCR and verified by sequencing. Finally, the over-recombinant expression vector was obtained and named ScPHR1-OE.

[0036] 2. Agrobacterium-mediated genetic transformation in Arabidopsis thaliana

[0037] The ScPHR1-OE recombinant expression vector was transformed into Agrobacterium EHA105 strain to obtain recombinant bacteria, which were then used for Arabidopsis genetic transformation experiments. The specific steps of the Agrobacterium-mediated transformation method are as follows: (1) Preparation of receptor materials Wild-type Arabidopsis thaliana (Col-0, Columbia type) with good growth status was selected as the transformation recipient.

[0038] (2) Prepare Agrobacterium-containing liquid

[0039] A single colony of Agrobacterium containing the target gene (recombinant single colony) was inoculated into a liquid culture medium, activated, and cultured to the logarithmic growth phase. After harvesting the bacterial suspension, the bacterial cells were resuspended in a permeate (1 / 2 MS powder + 10% sucrose + 400 μl / L Silwet-77) to achieve an OD600 value of 0.9, thus obtaining an Agrobacterium suspension.

[0040] (3) Infection

[0041] Immerse the inflorescence of Arabidopsis thaliana in Agrobacterium suspension, gently shake to ensure full contact between the inflorescence and the bacterial solution, and soak for 1 minute.

[0042] (4) Cultivation

[0043] Cover the plant with plastic wrap after the flower dipping treatment to maintain humidity, and incubate in the dark for 1 day. Then place it under normal incubation conditions. After 3 days, the plastic wrap can be removed, and the plant can continue to be incubated until it matures.

[0044] (5) Screening

[0045] The seeds of T0 generation Arabidopsis were collected, and the seeds were sown on 1 / 2MS medium containing kanamycin (40 mg / L) for growth screening to obtain positive plants. The positive Arabidopsis seedlings were transplanted into soil for continuous growth. After maturation, T1 generation seeds were collected for continuous screening, and the steps were repeated to obtain positive homozygous Arabidopsis seeds of T2-T3 generation.

[0046] A total of 3 positive homozygous Arabidopsis lines with significantly up-regulated expression were screened out (ScPHR1-OE-1, ScPHR1-OE-4, and ScPHR1-OE-8), which were used for further experimental verification. S

[0047] Example 4

[0048] Detection of transgenic Arabidopsis

[0049] Real-time fluorescence quantitative PCR (RT-qPCR) analysis was performed using qScPHR1-F / R specific primers (SEQ ID NO. 11-12), and the specific operation was performed according to the AceQ qPCR SYBR Green Master Mix (Vazyme) kit instructions. Sugarcane ScGAPDH gene and Arabidopsis AtActin2 gene were used as internal references, and the relative expression of the target gene was calculated by 2 -ΔΔCT - method. The reaction system was as follows: 10 μL 2x AceQ qPCR SYBR Green Master Mix, 0.4 μL forward primer (2 μM), 0.4 μL reverse primer (2 μM), 2 μL cDNA (200 ng / μL), and ddH2O was added to a total volume of 20 μL. The PCR reaction program was set as follows: 95°C pre-denaturation for 30 s; followed by 45 cycles of amplification: 95°C for 15 s, 60°C for 30 s; the melting curve analysis step was: 95°C for 15 s, 95°C for 60 s, 97°C for 1 s; finally, the temperature was reduced to 37°C and maintained for 30 s.

[0050] ScGAPDH The internal reference primers of the gene were as follows:qScGAPDH-F: 5'-CACGGCCACTGGAAGCA-3' (SEQ ID NO. 7); qScGAPDH-R: 5'-TCCTCAGGGTTCCTGATGCC-3' (SEQ ID NO. 8).

[0051] AtActin2 The internal reference primers of the qAtActin2-F: 5'-AGTGTCTGGATCGGTGGTTC-3' (SEQ ID NO. 9); qAtActin2-R: 5'-CCCCAGCTTTTTAAGCCTTT-3' (SEQ ID NO. 10).

[0052] qScPHR1-F: 5'-TTCCCAAAGCCTTGTAAC-3' (SEQ ID NO. 11); qScPHR1-R: 5'-TAGCCTGAATATGGTGCC-3' (SEQ ID NO. 12).

[0053] As shown in Table 1, the positive homozygous Arabidopsis lines were obtained by comparing the wild type Arabidopsis. ScPHR1 The gene expression amount is shown in Table 2 (the vertical coordinate is the expression amount). Figure 4

[0054] Figure 4 As shown in Table 3, the gene expression amount in the positive homozygous Arabidopsis lines is significantly higher than that of the wild type. ScPHR1

[0055] Example 5

[0056] ScPHR1 Verification of the function of the gene

[0057] The T3 generation of transgenic Arabidopsis seeds were first sterilized with 75% alcohol for 1 min, and then the alcohol was poured out. Then, 50% 84 disinfectant was used for sterilization for 2 min, and then poured out. Sterilized ddH2O was used for washing for 2 min, and then poured out, and the above steps were repeated 5 times. Finally, 0.1% agarose was added for resuspension, and the seeds were sown on 1 / 2MS medium containing 0.078 mM K2HPO4(LP, low-phosphorus medium) and 0.625 mM K2HPO4(CK, normal medium) after dark treatment for 2 d in a 4°C environment. The seed germination rate and leaf size were observed. The results are shown in Table 4. Figure 5~6

[0058] Figure 5 As shown in Table 5, the germination rate of wild type Arabidopsis seeds in low-phosphorus medium is lower than that of transgenic Arabidopsis.

[0059] Figure 6 As shown in Table 6, the leaf area of wild type Arabidopsis seeds in low-phosphorus medium is significantly lower than that of transgenic Arabidopsis.

[0060] Example 6

[0061] Determination of the resistance of transgenic Arabidopsis to Botrytis cinerea

[0062] The preserved Botrytis cinerea (​​​Botrytis. cinerea ) B. cinerea strain B05.10 was cultured on potato dextrose agar (PDA) medium at 22°C for 7 days, and conidia were collected and resuspended in ultrapure water. The conidial concentration was adjusted to 1 x 10 / mL. For whole-plant infection experiments, conidial suspension was sprayed evenly on the leaf surface of the plant using a hand-held sprayer. The inoculated plants were covered with a transparent cover to maintain a high humidity environment of more than 90% to promote fungal infection.

[0063] The leaves of B. cinerea-infected Arabidopsis plants were placed on the surface of sterile ultrapure water and treated by floating at room temperature for 3 h. Subsequently, the initial conductivity (E ) of the solution was measured using a DDSJ-308F conductivity meter (Lei Cun). Then, the sample was boiled for 30 min, and after cooling to room temperature, the final conductivity (E ) of the solution was recorded. The electrolyte leakage rate (%) was calculated according to the following formula: = (E / E ) x 100.

[0064] The results are shown in Figure 7 .

[0065] Figure 7 It is shown that, compared with the wild type control, the necrosis of the leaves of the transgenic material and the electrolyte leakage rate are significantly reduced after inoculation with B. cinerea, indicating that the transgenic Arabidopsis overexpressing ScPHR1 has enhanced resistance to B. cinerea.

[0066] From the above examples, it can be seen that a new sugarcane phosphate starvation response gene ScPHR1 is identified from sugarcane, and the function of resistance to low phosphorus and B. cinerea is analyzed. The tobacco subcellular localization results show that ScPHR1 is located in the nucleus. The results of transgenic Arabidopsis treated under low phosphorus conditions show that the overexpressed ScPHR1 can help Arabidopsis to improve the tolerance to low phosphate environment. The results of spraying the transgenic Arabidopsis with B. cinerea conidial liquid show that the overexpressed ScPHR1 can help Arabidopsis to improve the disease resistance to B. cinerea.

[0067] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. Sugarcane as shown in SEQ ID NO.1 ScPHR The application of gene 1 in improving plant tolerance to low phosphorus stress and resistance to gray mold is characterized by, The steps include: constructing a sugarcane containing the sugarcane shown in SEQ ID NO.

1. ScPHR 1. A recombinant expression vector for the gene was used to transform Agrobacterium tumefaciens into the recombinant expression vector. The recombinant bacteria carrying the recombinant expression vector were then used to infect plant plants. The method for constructing the recombinant expression vector is as follows: sugarcane ScPHR The gene was ligated to either the pBI121-GFP vector or the pCambia2300 vector; The Agrobacterium is GV3101 or EHA105; The plant in question is Arabidopsis thaliana or sugarcane.

2. The sugarcane shown in SEQ ID NO.1 ScPHR Application of gene 1 in the breeding of transgenic plants with low phosphorus stress resistance and gray mold resistance; the plant is Arabidopsis thaliana or sugarcane.

3. Sugarcane as shown in SEQ ID NO.1 ScPHR The application of a protein encoded by a gene in improving plant tolerance to low phosphorus stress and resistance to gray mold; the protein is shown in SEQ ID NO.

2.

4. Sugarcane as shown in SEQ ID NO.1 ScPHR The application of the protein encoded by gene 1 in the cultivation of transgenic plants with low phosphorus stress resistance and gray mold resistance; the plant is Arabidopsis thaliana or sugarcane.

5. A sugarcane expressing the form shown in SEQ ID NO.1 ScPHR A gene expression cassette, characterized in that, Including any of the following: (1) Recombinant expression vector: sugarcane ScPHR The gene was ligated to either the pBI121-GFP vector or the pCambia2300 vector; (2) Recombinant bacteria: sugarcane ScPHR 1 gene or recombinant expression vector was transferred into Agrobacterium GV3101 or EHA105.

6. The application of the expression cassette according to claim 5 in improving plant tolerance to low phosphorus stress and resistance to gray mold; wherein the plant is Arabidopsis thaliana or sugarcane.

7. The application of the expression cassette according to claim 5 in the cultivation of transgenic plants with low phosphorus stress resistance and gray mold resistance; wherein the plant is Arabidopsis thaliana or sugarcane.

8. A method for detecting the sugarcane shown in SEQ ID NO.1 ScPHR The application of reagents for measuring gene expression levels in plants in evaluating plant resilience to low phosphorus stress; the reagents are primer pairs; the primer pairs are shown in SEQ ID NO. 11~12.

9. A method for detecting the sugarcane shown in SEQ ID NO.1 ScPHR The application of reagents for measuring gene expression levels in plants in evaluating plant resistance to gray mold; the reagents are primer pairs; the primer pairs are shown in SEQ ID NO. 11~12.

10. A method for improving plant tolerance to low phosphorus stress and resistance to gray mold, characterized in that, The sugarcane shown in SEQ ID NO.1 ScPHR 1. Overexpression of the gene in plants; or construction of a sugarcane containing the gene shown in SEQ ID NO.

1. ScPHR 1. A recombinant expression vector for the gene was used to transform Agrobacterium tumefaciens into the recombinant expression vector. The recombinant bacteria carrying the recombinant expression vector were then used to infect plant plants. The Agrobacterium is GV3101 or EHA105; The plant in question is Arabidopsis thaliana or sugarcane.

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