Application of rice monosaccharide transporter gene OsPLT1 and transporter of rice monosaccharide transporter gene OsPLT1 in regulation and control of growth period of rice and amplification primer

Abstract

The invention belongs to the field of biological genes, and discloses a rice monosaccharide transporter gene OsPLT1 and application of a transporter of the rice monosaccharide transporter gene OsPLT1 in regulation and control of the growth period of rice, and the nucleotide sequence of the rice monosaccharide transporter gene OsPLT1 is shown as SEQ ID NO: 1. The invention further discloses an amplification primer for the rice monosaccharide transporter gene OsPLT1. The invention provides a novel application of a rice OsPLT1 gene and an encoding protein thereof in regulating and controlling the growth period of rice. When the OsPLT1 is knocked out from the wild type ZH11 through a CRISPR / Cas9 method, the growth process of the rice can be accelerated, and the yield of the rice is not influenced, so that the OsPLT1 gene can be used for regulating the growth period of the plant and improving the annual yield of the plant, and has relatively high economic and application values.

Description

Technical Field

[0001] The present invention belongs to the field of biological genes, and in particular relates to a rice monosaccharide transfection gene OsPLT1 and its transporters in shortening the growing period of rice. Background Art

[0002] There are three major sugar transporter families in rice, including sucrose transporters (SUT), sugar efflux transporters (SWEET) and monosaccharide transporters (MST), covering a total of 92 genes. Among them, the functions of MST proteins are less reported. According to the different transport substrates, they can be divided into plasma membrane monosaccharide transporters (STPs), inositol transporters (INTs), tonoplast monosaccharide transporters (TMTs), tonoplast glucose transporters (VGTs), plastid monosaccharide transporters (GMTs), early response dehydration-like proteins (ERDs) and polyol transporters (PLTs). There are 6 polyol transport genes in Arabidopsis ( PLT1-6 : At2g16120 , At2g16130 , At2g18480 , At2g20780 , At3g18830 , At4g36670 ). Studies have shown that these genes, in addition to transporting alcohols, are also involved in the transport of monosaccharides such as glucose and xylose. AtPLT1 ( At2g16120 )and AtPLT2 ( At2g16130 ) is a transporter of xylitol and fructose, mainly expressed in pollen and young xylem; AtPLT5 ( At4g36670 ) transports sorbitol, xylitol, erythritol, and xylose, and is most strongly expressed in roots, leaf vascular tissues, and specific floral organs. PLT Related studies have shown that its transport substrates are not limited to sugar alcohols, but its research mainly focuses on tissue-specific expression, subcellular localization and transport substrates. AtPLT1 / 2 / 5 The mutant materials of these genes have no significant difference from the wild type, so the biological functions of these genes are not clear. PLT The biological functions and mechanisms of action in plants are also rarely reported. PLT The relevant research is a blank area. Summary of the invention

[0003] The technical problem to be solved by the present invention is to overcome the deficiencies and defects mentioned in the above background technology and provide a rice monosaccharide transporter gene OsPLT1 Application of its transporter in regulating rice growth period and amplification primers.

[0004] In order to solve the above technical problems, the technical solution proposed by the present invention is: A rice monosaccharide transporter gene OsPLT1 Application in regulating the growth period of rice, the rice monosaccharide transporter gene OsPLT1 The nucleotide sequence is shown in SEQ ID NO:1.

[0005] Based on a general inventive concept, the present invention also provides a rice monosaccharide transporter gene OsPLT1 The encoded transporter is used in regulating the growth period of rice, and the amino acid sequence of the transporter is shown in SEQ ID NO: 2.

[0006] The above application, preferably, is to knock out in wild-type rice by CRISPR / Cas9 method OsPLT1, The short-cultivation period rice obtained.

[0007] The above application is preferably from OsPLT1 Selecting monosaccharide transporter genes from gDNA sequences OsPLT1 The target sequence was edited, and the Cas9-OsPLT1 vector was constructed. Then it was transformed into Agrobacterium by heat shock transformation method, and finally cloned into rice by genetic transformation for expression. OsPLT1 Gene-edited transgenic lines.

[0008] In the above application, preferably, the monosaccharide transporter gene OsPLT1 The edited target sequences are SEQ ID NO:3 and SEQ ID NO:4.

[0009] The above application, preferably, the OsPLT1 The gene-edited transgenic strains were identified by PCR amplification, with the identification primers being Cas9::OsPLT1, the upstream primer sequence being 5'-CGTGTTAGCTCGGATCTGTA -3' (as shown in SEQ ID NO: 5), and the downstream primer sequence being 5'-GGCGAAGTTGGAGACGTA -3' (as shown in SEQ ID NO: 6).

[0010] Based on a general inventive concept, the present invention also provides a method for producing a monosaccharide transporter gene in rice. OsPLT1 Preferably, the amplification primer is G::OsPLT1, and the sequences of the forward primer and the reverse primer are shown in SEQ ID NO: 7 and SEQ ID NO: 8, respectively. The amplification primer G:: OsPLT1 is used to amplify the monosaccharide transport gene of rice. OsPLT1 Specific CDS sequence, suitable for verifying monosaccharide transporter genes OsPLT1 Expression profile of the rice monosaccharide transporter gene OsPLT1 The nucleotide sequence is shown in SEQ ID NO:1.

[0011] Based on a general inventive concept, the present invention also provides a method for producing a monosaccharide transporter gene in rice. OsPLT1 The amplification primer is OE::OsPLT1-GFP, and the sequences of the forward primer and the reverse primer are shown in SEQ ID NO: 9 and SEQ ID NO: 10, respectively. The amplification primer OE::OsPLT1-GFP is used to amplify the CDS sequence of the rice monosaccharide transport gene OsPLT1, and is suitable for the subcellular localization experiment of the monosaccharide transporter OsPLT1 in rice.

[0012] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides rice OsPLT1 New use of genes and their encoded proteins in regulating rice growth period; knocked out in wild-type ZH11 by CRISPR / Cas9 method OsPLT1 It can accelerate the growth process of rice without affecting the yield of rice. OsPLT1 Genes can be used to regulate the growth period of plants and increase the annual yield of plants, and have high economic and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 is obtained in Example 1 of the present invention OsPLT1 The spatiotemporal expression pattern results of (where P: panicle, YP: young panicle, LB: leaf blade, LS: leaf sheath, S: stem, N: node, SB: stem base); Figure 2 is the subcellular localization result of OsPLT1 obtained in Example 1 of the present invention; Figure 3 is obtained in Example 2 of the present invention OsPLT1 Two different mutation types; Figure 4 is obtained in Example 2 of the present invention OsPLT1 The tillering stage phenotypes of mutants and wild type; Figure 5 is obtained in Example 2 of the present invention OsPLT1 The hydroponic seedling phenotypes of mutants and wild type; Figure 6 is obtained in Example 2 of the present invention OsPLT1 The heading phenotype and heading rate statistics of mutants and wild type; Figure 7 is obtained in Example 2 of the present invention OsPLT1 The mutant and wild-type ear development phenotypes; Figure 8The maturity obtained in Example 2 of the present invention OsPLT1 Statistics of yield-related agronomic traits of mutants and wild type. DETAILED DESCRIPTION

[0014] In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively and meticulously below in conjunction with the accompanying drawings and preferred embodiments of the present invention, but the protection scope of the present invention is not limited to the following specific embodiments.

[0015] Unless otherwise defined, all professional terms used below have the same meanings as those generally understood by those skilled in the art. The professional terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention.

[0016] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0017] Embodiment 1: A rice monosaccharide transporter gene OsPLT1 The nucleotide sequence is shown in SEQ ID NO: 1; the amino acid sequence of the transporter encoded by it is shown in SEQ ID NO: 2.

[0018] SEQ ID NO: 1: SEQ ID NO: 2: MTKDDAVPVAVAPAKRPPINKYAFGCALLASMNSVLLGYDISVMSGAQIFMKEDLKITDTQIEILAGVINIYSLFGSLAAGMTSDWLGRRYTMVLAAAIFFTGALLMGLAPNYAFLMAGRFVAGIGVGY ALMIAPVYTAEVAPTSARGFLTSFPEVFNNSGILLGYVSNFAFARLPVHLSWRAMFLVGAVPPIFLGIAVLAMPESPRWLVMRGRIEDARRVLLKTSDSPDEAEDRLLDIKKAVGIPEDASDGEDVVAIV RANKASQGEGVWKELLLNPTRPVRRMLVAGLGLMFIQQATGVDCVVMYSPRVFERAGIKSKTNSLGASMAVGVCKTFFIPIATLLLDRVGRRPLLLASGGGMAIFLFTLATSLLMMDRRPEGEAKALGA ISIAAMLSFVASFASGLGPVAWVYTSEIYPVRLRAQAAAIGTGLNRLMSGATTMSFLSLSNAITIAGSFYLYASIAAAGWVFMYFFLPETKGKSLEDTVKLFGKDTDDDDDVDTSRHERKRSTELSAQH* Rice monosaccharide transporter gene OsPLT1 The CRISPR-Cas9 technology and its transporter were discovered in rice using plant molecular genetics and physiological and biochemical methods. After research, we also found that the mutation of its gene significantly shortened the growth period of rice but did not affect the yield, which provides a practical basis for enhancing the transport and distribution capacity of photosynthetic products through genetic improvement of varieties and continuously improving the annual yield of rice.

[0019] 1. Spatiotemporal expression patterns of rice monosaccharide transporter genes To obtain the rice monosaccharide transporter gene OsPLT1To investigate the expression pattern of OsPLT1, RNA was extracted from various parts of rice ZH11 at different growth stages, with three replicates for each sample. Total RNA was extracted using TRIzol, and the first-strand cDNA was synthesized using the Hiscript II Q RT SuperMix kit (Vazyme). Real-time fluorescence quantitative PCR analysis was performed on a StepOnePlus instrument using the qPCR premix ChamQUniversal SYBR qPCR Master Mix (Vazyme). The PCR amplification primers were G::OsPLT1, and its forward and reverse primers were 5'- CTTCCTCTCGCTCTCCAA-3' (as shown in SEQ ID NO: 7) and 5'- TCATCGTCGTCGTCTGT -3' (as shown in SEQ ID NO: 8), respectively. Ubiquitin was used as an internal standard with primers 5'- GCTCCGTGGCGGTATCAT -3' (as shown in SEQ ID NO: 11) and 5'- CGGCAGTTGACAGCCCTAG -3' (as shown in SEQ ID NO: 12).

[0020] The specific steps are as follows: Total RNA extraction: 1. After tissue sampling, freeze in liquid nitrogen and grind into powder. Take 0.1 g sample and add it to a 1.5 mL centrifuge tube. Add 1 mL TRIzol, mix quickly, and let stand at room temperature for 5 min. 2. Add 200 µL chloroform to the centrifuge tube, shake vigorously for 30 s, and let stand at room temperature (25°C) for 10 min. 3. Centrifuge at 4°C 12000 rpm for 10 min. 4. Carefully transfer 500 µL of supernatant to a new 1.5 mL centrifuge tube. 5. Add an equal volume of isopropanol, gently invert to mix, let stand at room temperature (25°C) for 10 min. Centrifuge at 4°C 12000 rpm for 10 min, and discard the supernatant. 6. Add 1 mL 75% ethanol and gently invert to suspend the precipitate. 7. Centrifuge at 4°C 8000 rpm for 5 min, and discard the supernatant. 8. Repeat steps 6 and 7, blow the RNA precipitate to a translucent gel in a clean bench, add 40 µL DEPC water to dissolve RNA.

[0021] RNA quality detection and concentration determination: 1. Electrophoresis detection of RNA integrity: 1% agarose gel fast electrophoresis, detection of RNA molecular integrity, observation of 18S and 28S bands, 28S band brightness is about 2 times the brightness of 18S band, indicating that RNA integrity is good; 2. Nucleic acid analyzer detection of RNA purity and concentration: DEPC water is used to read BLANK, NanoDrop to determine RNA concentration. The absorption peak of nucleic acid is at 260 nm, the maximum absorption peak of protein is at 280 nm, and OD 260 / 280 The purity of RNA samples between 1.8-2.0 meets the standard and can be used for subsequent fluorescence quantitative experiments.

[0022] Synthesis of the first strand of cDNA: Total RNA was used to synthesize the first strand of cDNA using the reverse transcription kit Hiscript II Q RT SuperMix. The total amount of cDNA for each sample was 1 µg. cDNA was stored at -20°C for future use. The reverse transcription system was as follows: Total RNA (1 µg), 4 × gDNA wiper Mix 4 µL, 5 × HiScript® II qRT SuperMix II 4µL, and RNA-free water was added to 20 µL. Reaction conditions: incubate at 42°C for 15 min, and inactivate at 85°C for 5 s. After the reaction was completed, an equal volume of RNA-free water was added to dilute it by half, and after mixing, it was stored in a refrigerator at -20°C for future use.

[0023] Real-time fluorescence quantitative PCR primer design and specificity analysis: qPCR primers were designed according to the full length of gene cDNA using the software primer5. Primers with good specificity and no complementary bases were selected and sent to Qingke Biotechnology Co., Ltd. for synthesis to obtain the amplification primer G::OsPLT1. The cDNA of rice at different stages was used as a template, and the primer specificity was detected by agarose gel electrophoresis. The reaction system was as follows: cDNA 1 µL, G::OsPLT1 forward primer (10 µM) 0.5 µL, G::OsPLT1 reverse primer (10µM) 0.5 µL, 1.1×T3 Super PCR Mix (Qingke Biotechnology) 18 µL. Reaction conditions: 95℃, 3 min; 95℃, 10 s; 55℃, 10 s; 72℃, 10 s; 36 cycles in total, 72℃, 5 min.

[0024] After the primers for fluorescence quantitative PCR were detected correctly, the first strand of cDNA synthesized by reverse transcription was used as a template and quantified by a two-step method on a Roche LightCycler®96 real-time fluorescence quantitative PCR instrument. Three technical replicates were set for each gene and each sample. The reaction system was as follows: 1 µL cDNA, 0.5 µL G::OsPLT1 forward primer (10 µM), 0.5 µL G::OsPLT1 reverse primer (10 µM), 10 µL 2 × ChamQ Universal SYBR qPCR Master Mix, and 8 µL ddH2O. Reaction conditions: (1) Pre-denaturation stage: 95°C, 10 min; (2) Amplification stage: 95°C, 10 s, 60°C, 30 s, for a total of 40 cycles; (3) Melting curve stage: 96°C 15 s, 60°C 60 s, and 95°C 15 s.

[0025] The results are as follows Figure 1 As shown, OsPLT1 It is expressed in all tissues at all stages, with relatively high expression in leaves and leaf sheaths during the tillering stage, leaves, leaf sheaths and stems during the jointing stage, stems, nodes and ears during the booting and heading stages, and ears during the grain filling stage.

[0026] 2. Rice monosaccharide transporter gene OsPLT1 Subcellular localization For research OsPLT1 To enhance the cell specificity of expression, we constructed a transformation vector carrying OsPLT1-GFP. OsPLT1 The CDS sequence of OE::OsPLT1-GFP (excluding the terminator) was amplified using the forward and reverse primers 5'- Gagctcggtacccgg ggatcc ATGACCAAGGACGACGCCG-3' (as shown in SEQ ID NO: 9, Bm I recognition site is italicized) and 5'-CTTCTCTTTGCCCAT GTCGAC GTGCTGAGCGCTCAGCTC -3' (as shown in SEQ ID NO: 10, Sal I recognition sites are italicized). Use Bm I and Sal The fragment was cut by endonuclease I, and the amplified fragment was cloned into the pCAMBIA1300-GFP vector to generate the OsPLT1-GFP vector, which was further transformed into rice protoplasts by a PEG-mediated method.

[0027] The specific steps are as follows: shake the OsPLT1-GFP bacterial suspension, extract the OsPLT1-GFP plasmid using the Solebow Plasmid Extraction Kit and concentrate the plasmid. The concentration steps are as follows: (1) mix all plasmids of the same gene into one tube; (2) freeze the centrifuge tube containing the plasmid at -80℃; (3) seal the centrifuge tube with sealing film and poke two holes in the tube cap with a needle; (4) freeze dry the plasmid using a freeze dryer; (5) add ddH2O to adjust the concentration to ≥1 µg and store at -20℃ for later use.

[0028] Preparation of rice protoplasts: (1) Cultivate rice ZH11 in 1 / 2MS and grow it in the dark for 11-14 days. Select 15 yellow flower seedlings with good growth status and cut them into thin strips with disposable sterile blades. The thinner the better. (2) Place them in a culture dish containing 10 mL of enzymatic solution, evacuate for 5 min, and enzymolyze them in a shaker in the dark for 4-5 h (28℃, 45 rpm). (3) Filter the enzymatic solution into a 10 mL round-bottom centrifuge tube using a 150-mesh sieve. (4) Centrifuge at 200 g for 1 min at 4℃, discard the supernatant, gently add 1 mL of W5 (154 mM NaCl; 125 mM CaCl2; 5 mM KCl; 2 mM MES) along the wall, and gently resuspend the protoplasts until there is no precipitation at the bottom. (5) Incubate on ice for 40 min. (6) Remove the supernatant, add 2 mL of W5 and gently invert to mix. (7) Cut off the tip of the yellow pipette and pipette 12 10 µL of the mixed sample is gently transferred into a hemocytometer and placed under a microscope for counting; (8) The remaining protoplasts are incubated on ice for 40 min and the supernatant is discarded.

[0029] PEG-mediated protoplast transformation: Estimate the number of protoplasts based on the protoplast concentration and add MMG (0.6 M Mannitol; 15 mM MgCl2; 4 mM MES) to make the final protoplast concentration approximately 2×10 5 / mL; prepare the following reaction solution in a 1.5mL round-bottom centrifuge tube: 150 µL protoplasts, 15 µg GFP plasmid (1.0 µg / µL), mix evenly; then add 165 µL PEG transformation solution, gently invert up and down several times to promote the fusion of PEG and protoplasts, let stand at room temperature for 5 minutes, add 1 mL W5 to terminate the reaction; centrifuge at 200 g for 5 min, remove the supernatant, resuspend the protoplasts with 200 µL W5, and culture overnight at 22℃ in the dark; observe the GFP signal of the protoplasts using a laser confocal microscope.

[0030] The results are as follows Figure 2 As shown, rice monosaccharide transporter gene OsPLT1 Localized on the cell membrane of rice protoplasts.

[0031] Embodiment 2: Rice monosaccharide transport gene of the present invention OsPLT1 Application in the cultivation of short-growing rice, including OsPLT1 Construction of mutant materials, phenotypes at different growth and development stages, and yield factor analysis.

[0032] 1. OsPLT1 Obtaining gene knockout materials In order to verify the rice monosaccharide transport gene transporter of the present invention OsPLT1 In rice, we used CRISPR-Cas9 gene editing technology to OsPLT1 The Cas9 / g RNA vector was constructed with target sequence 1: 5'-CCCATCAACAAGTACGCCTTCGG-3' (as shown in SEQ ID NO: 3) and target sequence 2: 5'-GGGCATGACGTCCGACTGGCTGG-3' (as shown in SEQ ID NO: 4), and Agrobacterium tumefaciens GV3101 was transformed. The positive strain was used for genetic transformation of wild-type rice ZH11 to obtain OsPLT1 Gene-edited transgenic strains. The transgenic strains were identified by PCR amplification, with the primers Cas9::OsPLT1, the upstream primer sequence: 5'-CGTGTTAGCTCGGATCTGTA -3' (as shown in SEQ ID NO: 5), and the downstream primer sequence: 5'-GGCGAAGTTGGAGACGTA -3' (as shown in SEQ ID NO: 6). After identification, two knockout strains with different mutation types were obtained. plt1-1 , plt1-2 .

[0033] The results are as follows Figure 3 As shown, plt1-1 A single base C is deleted near target site 1 and target site 2, resulting in OsPLT1 The gene encoding 31 amino acids terminates prematurely; plt1-2 Only a single T is inserted near target site 1, resulting in OsPLT1 The gene encoding 39 amino acids terminates prematurely.

[0034] 2.Os PLT1 Effects of gene knockout on rice growth To evaluate Os PLT1 The effects of gene knockout on rice germination and seedling growth were compared between wild-type ZH11 and plt1Seed germination and seedling growth phenotypes of mutants. Seeds were soaked in 5000-fold diluted prochloraz for 16 h, rinsed with running water, and then soaked in sterile distilled water overnight at 37°C. Seeds were sown on completely moistened filter paper and cultured in an incubator at 28°C with 12 h of light and 12 h of darkness. Photos were taken from the first to the fourth day, and the germination rate was counted. After the seeds turned white, they were moved to hydroponics and cultured for 14 days to observe the growth of ZH11 and plt1 The growth of mutants, and the plant height and dry weight were calculated. The values ​​are the mean ± standard deviation of 10 repeated experiments, and the P value is the statistical test of Student's t -test analysis* P <0.05,** P <0.01.

[0035] The results are as follows Figure 4 As shown, compared with the wild type ZH11 grown in the field, plt1 The mutant had no effect on seed germination and seedling growth.

[0036] To evaluate Os PLT1 Effects of gene knockout on tillering period of rice, field cultivated wild type (ZH11), plt1 Mutants ( plt1-1 , plt1-2 ), analysis of Os PLT1 The tillering phenotype of the genetic material was analyzed, and the tillering number and plant height were counted. The values ​​are the mean ± SD of 15 repeated experiments, and the P value is the statistical test of significant difference Student's t -test analysis* P <0.05,** P <0.01.

[0037] The results are as follows Figure 5 As shown, compared with ZH11, plt1 The tiller number and plant height of the mutant increased significantly.

[0038] To evaluate Os PLT1 The effects of gene knockout on the reproductive growth period of rice were observed from the 57th day after sowing. plt1 Mutants ( plt1-1 , plt1-2 The heading situation of the materials in the plot was counted every three days until the wild type (ZH11), plt1 Mutants ( plt1-1 , plt1-2 ) All materials reached the heading stage and OsPLT1 The phenotypes of genetic materials at heading stage were recorded by photographing; starting 30 days after sowing, samples of wild type (ZH11), plt1 Mutants ( plt1-1 , plt1-2 ) Materials: Take a whole rice plant, carefully peel off the leaves of the plant, and take photos of the tip of the main stem.

[0039] The results are as follows Figure 6 and Figure 7 As shown, compared with wild-type ZH11, plt1 The mutant phenotype showed that the heading time was advanced by nearly 8 days. plt1 The ear development process of the mutant was significantly advanced.

[0040] 3.Os PLT1 Effects of gene knockout on agronomic traits related to rice yield Field cultivation of wild type ZH11, plt1 Mutants ( plt1-1 , plt1-2 ) materials, and the yield-related agronomic traits were statistically analyzed after maturity. Compared with the wild type ZH11, plt1 There were no differences in the main ear length, number of grains per ear, number of effective tillers, thousand-grain weight, fruit setting rate, and yield per plant among the mutants ( Figure 8 ). The values ​​are the mean ± standard deviation of 15 repeated experiments, and the P value is a significant difference statistical test Student's t-test analysis *P<0.05, **P<0.01.

[0041] In summary, the rice monosaccharide transporter gene of the present invention OsPLT1 , located in the plasma membrane of rice protoplasts, its knockout mutation leads to a significant advancement of the growth period of rice, by nearly 8 days, which is not affected by the germination rate and OsPLT1 The gene knockout mutation does not affect rice yield.

Claims

1. A rice monosaccharide transporter gene OsPLT1 The application in regulating the growth period of rice is characterized in that: The rice monosaccharide transporter gene OsPLT1 The nucleotide sequence is shown in SEQ ID NO:

1.

2. A monosaccharide transporter gene from rice OsPLT1 The use of the encoded transporter in regulating the growth period of rice is characterized in that: The amino acid sequence of the transporter is shown in SEQ ID NO:

2.

3. The use according to claim 1 or 2, characterized in that: Knockout in wild-type rice by CRISPR / Cas9 approach OsPLT1, The short-cultivation period rice obtained.

4. The use according to claim 1 or 2, characterized in that: from OsPLT1 Selecting monosaccharide transporter genes from gDNA sequences OsPLT1 The target sequence was edited, and the Cas9-OsPLT1 vector was constructed. Then it was transformed into Agrobacterium by heat shock transformation method, and finally cloned into rice by genetic transformation for expression. OsPLT1 Gene-edited transgenic lines.

5. The use according to claim 4, characterized in that The monosaccharide transporter gene OsPLT1 The edited target sequences are SEQ ID NO:3 and SEQ ID NO:

4.

6. The use according to claim 4, characterized in that Said OsPLT1 The gene-edited transgenic strains were identified by PCR amplification, the identification primers were Cas9::OsPLT1, the upstream primer sequence was as shown in SEQ ID NO: 5, and the downstream primer sequence was as shown in SEQ ID NO:

6.

7. A monosaccharide transporter gene for rice OsPLT1 The amplification primer is characterized in that The amplification primer is G::OsPLT1, and the sequences of the forward primer and the reverse primer are shown in SEQ ID NO: 7 and SEQ ID NO: 8, respectively. OsPLT1 The nucleotide sequence is shown in SEQ ID NO:

1.

8. A monosaccharide transporter gene for rice OsPLT1 The amplification primer is characterized in that The amplification primer is OE::OsPLT1-GFP, and the sequences of the forward primer and the reverse primer are shown in SEQ ID NO: 9 and SEQ ID NO: 10, respectively. OsPLT1 The nucleotide sequence is shown in SEQ ID NO:1.

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