CN114752605B

CN114752605B - Rice OsOFP22 s Gene and method for increasing grain length, thousand grain weight and improving amylose content of rice by using same

Info

Publication number: CN114752605B
Application number: CN202210594164.6A
Authority: CN
Inventors: 李钱峰; 白皇; 刘巧泉; 任新雨; 张昌泉; 张�林; 陆陈雅
Original assignee: Yangzhou University
Current assignee: Yangzhou University
Priority date: 2022-05-27
Filing date: 2022-05-27
Publication date: 2023-07-21
Anticipated expiration: 2042-05-27
Also published as: CN114752605A

Abstract

The invention discloses a rice OsOFP22 ^s Gene and method for improving grain length, thousand grain weight and amylose content of rice by using same, osOFP22 of rice ^s The gene is highly expressed in seeds, can specifically improve rice grain related characters including grain length, thousand grain weight and amylose content, has no obvious influence on other main agronomic characters of rice plants, and over-expresses OsOFP22 in rice by utilizing a genetic engineering technology ^s Can obviously improve the thousand grain weight of rice and improve the grain shape and the cooking taste quality; by comparison, osOFP22 ^s The over-expressed rice is superior to the control in grain length and thousand grain weight, the amylose content is obviously reduced compared with the control, the cooking taste quality is obviously improved, and OsOFP22 ^s The gene has good application prospect in high-yield and high-quality breeding of rice.

Description

Rice OsOFP22 s Gene and method for increasing grain length, thousand grain weight and improving amylose content of rice by using same

Technical Field

The invention belongs to the technical field of plant genetic engineering, and in particular relates to a gene OsOFP22 for improving rice grain length, thousand seed weight and amylose content ^s And uses thereof, and also relates to polypeptide sequences encoded by the genes.

Background

Rice (Oryza sativa L.) is a staple food of more than half of the world population, and China is the first major country of global rice yield and consumption. With the growth of population and the gradual decrease of cultivated land area, the improvement of the yield per unit area of rice becomes the most important way for improving the total yield of grains in order to ensure the national grain safety. Meanwhile, with the improvement of the living standard of people and the opening of the rice market, rice is widely traded as commodity, and people also put forward higher and higher requirements on the quality of rice appearance, cooking taste and the like. However, the rice breeding in China always takes high yield as a main target, the quality improvement progress is relatively slow, so that the quality rice in China has fewer varieties and has weak competitiveness in the international rice market. Therefore, it is important to further improve the rice quality on the basis of ensuring the yield. At present, the breeding of new rice varieties with high quality and high yield has become the most important goal of rice breeding in China.

The grain shape of the rice is not only an important factor for determining the yield of the rice, but also an important index for measuring the appearance, processing and other qualities of the rice. In addition, the grain shape is less affected by the environment, and the genetic effect is obvious. Therefore, improving the grain shape of rice is an effective way for realizing high-yield and high-quality breeding of rice. In order to accelerate the breeding process of rice grain shape improvement, besides adopting a conventional breeding method, more important is to discover more available grain shape genes, perfect a molecular network for regulating and controlling rice grain shapes, construct a high-efficiency molecular breeding technical system and cultivate new rice varieties with specific grain shapes through a molecular design breeding strategy.

The main indexes of the rice grain shape property comprise grain length, grain width, grain thickness, length-width ratio and the like. Wherein grain length not only affects the appearance and milling quality of rice, but also has a significant positive correlation with thousand grain weight of rice. Before, the gene GS9 for specifically regulating the grain shape of the rice is separated and cloned by a map cloning method, the grain of the rice material is knocked out, the appearance quality is improved, and the main agronomic characters such as plant height, thousand grain weight and the like are not obviously influenced. The grain shape property is obviously improved by knocking out the GS9 gene in different rice varieties. Selecting a protein which interacts with GS9 protein in a rice seed cDNA library by using the GS9 protein as a bait, and selecting an oval protein family member OsOFP22 ^s Can directly interact with GS9 and further verify in plants by using luciferase complementation assayInteraction of two proteins.

Disclosure of Invention

The invention aims at the problems and provides a gene OsOFP22 for improving rice grain length, thousand seed weight and amylose content ^s And applications thereof.

The aim of the invention is realized by the following technical scheme: rice OsOFP22 ^s Gene, osOFP22 ^s The gene is located on chromosome 1 of rice, and the gene number is Os05g0477200 (NCBI number), LOC_Os05g39950 (MSU number). OsOFP22 ^s The CDS full length of the gene coding region 1155bp, no intron exists, and the OsOFP22 ^s The coding region sequence of the gene is shown in SEQ ID NO.1 (ATGGGCCGGCGGAAGTTCAGGCTCTCCGACATGATGCCCAACGCGTGGTTCTACAAGCTCCGCGACATGCGCGCGCGGGGCGGCCGCGGTGCAACTGCGATGCAGCCGCCGTCGTCGTCGTCGTTGATGAGGGGGAGCAGGGCGGCGCAGCAGCAGGCGGGCACGTGGAGGCTGGGGACGTCGTCGTCGTCGTCGTCGTTGCTGCCGCACAGGGCGTCGTACTACTACACCACCCGGGACAGGGAGGTCCCGCCGCTGCCGCCGCCGCCACCGCCGAGGGGCGTGGATGATCAATTCCCTTCCCTCACGCTGTCGCCGCCGTTGCCGACGAGGAACAGCAGGAGGCGGCACAGGGTTGGGAGATTTGGTTCGACGGAGATGGATGGCGGCGAGCTCGTACTAGCGCCGTCCGACGACCACGACGGCTGCAGCCACCAGGAGCCGCCAGTGGCCGATGCGTCCGGGAGCTCCCGGTGCCGTCGCGACATGTTCATCGGGAGAGATGGCGGCCGGGGCGTGGAGTTCCGGCGCCGGGCGACGACGGTGGATGGTCCTGAGGAGGACGCCGCCGTCGATGTCAAGGTGATCACGTCGGACGCGGACATAATCATCGACCTCGGCGCTGACGACGACGACGACACGCCGGAGAGGGTGCTCCGGCCTGTCGTGACCAGGCCCGCGAGGAGGGAGCTCGACTGGTGCGAGCCGGCGGAGGTGAAGCACGTCGACCTCGCCGAGCTGATGACACCGAGAGCGAGCTCTGCCTCTGCCTCCTCGGAGAAGAGCATCAGCACGGGCAAGCCGAGGCGTTCGTCCGTGTCGTCTCGACGCCGCCTCAAGACGCGCACCAACAGCCCGCGCCTCGCCGCGTGCAGGAAAGGCAAGCCGACGGCGCGGGCAACGACGACGACGCCGACGCAGCCGCCGCTCGCGCACAGCTTCGCGGTGGTGAAGACGTCGTCGGACCCGAGGAGGGACTTCCTCGAGTCCATGGAGGAGATGATCGCCGAGAACGGCATCCGCGACGCCGGCGACCTGGAGGACCTCCTCGCCTGCTACCTCTCCCTCAACTCCGGCGAGTACCATGACCTCATCGTCGAGGTGTTCGAGCAGGTCTGGACCGGCCTCGCCGCTGCCTGTGGCGTCATGCCATGA); osOFP22 ^s 384 amino acids encoded by the gene, osOFP22 ^s The coded amino acid sequence is shown in SEQ ID NO.2 (MGRRKFRLSDMMPNAWFYKLRDMRARGGRGATAMQPPSSSSLMRGSRAAQQQAGTWRLGTSSSSSSLLPHRASYYYTTRDREVPPLPPPPPPRGVDDQFPSLTLSPPLPTRNSRRRHRV)GRFGSTEMDGGELVLAPSDDHDGCSHQEPPVADASGSSRCRRDMFIGRDGGRGVEFRRRATTVDGPEEDAAVDVKVITSDADIIIDLGADDDDDTPERVLRPVVTRPARRELDWCEPAEVKHVDLAELMTPRASSASASSEKSISTGKPRRSSVSSRRRLKTRTNSPRLAACRKGKPTARATTTTPTQPPLAHSFAVVKTSSDPRRDFLESMEEMIAENGIRDAGDLEDLLACYLSLNSGEYHDLIVEVFEQVWTGLAAACGVMP)。

The rice OsOFP22 ^s Gene overexpression recombinant vector pActin-OFP22 ^s -3Flag, characterized in that the pActin-OFP22 ^s -3Flag contains OsOFP22 ^s The gene and the vector are plant binary expression vector pActin-3Flag.

The first strand of cDNA obtained by reverse transcription was amplified by PCR to give a target band. After amplification, the product containing the target gene is ligated to an intermediate vector, and the ligation product is transformed into E.coli competent cells. Shaking bacteria, selecting single colony, enzyme cutting, identifying and sequencing. If the sequencing result is correct, extracting plasmid, cutting out target gene and connecting it into final expression vector pActin-OFP22 ^s -3Flag。

Further, the overexpression recombinant vector pActin-OFP22 ^s The preparation method of the-3 Flag comprises the following steps:

the plant binary expression vector pCAMBIA1300 is digested with Sam I and Xba I, and the digested vector is digested with homologous recombination enzymes to obtain the vector containing OsOFP22 ^s Ligation of PCR amplified products of full-length cDNA of the gene.

Preferably, the polypeptide comprises OsOFP22 ^s The PCR amplification method of the full-length cDNA of the gene is as follows:

the primer sequences were as follows using wild type RNA as template:

sequence name	Sequence(s)	Sequence numbering
			OFP22 ^s -OE-F	5-TAGGTAGAAGAGGTACCCGGGCTCTGGCCTGGCCCCCCA-3	SEQ ID NO.3
OFP22 ^s -OE-R	5-GTAATCTCCGTCGACTCTAGATGGCATGACGCCACAGGC-3	SEQ ID NO.4

。

Further, the rice OsOFP22 ^s The gene is applied to the aspects of increasing the grain length, thousand grain weight and improving the amylose content of rice.

Preferably, the method of application is as follows: with said rice OsOFP22 ^s The gene sequence is used for constructing a recombinant vector which can be overexpressed in rice, transforming agrobacterium, transferring the vector into rice callus by using an agrobacterium-mediated rice genetic transformation method, and obtaining rice plants after resistance screening and tissue culture.

Preferably, the recombinant vector overexpressed in rice is the overexpression recombinant vector pActin-OFP22 ^s -3Flag。

Preferably, the application method is as follows:

(1) Constructing engineering bacteria: pActin-OFP22 ^s The 3Flag vector was transformed into Agrobacterium strain EHA105 by electric shock, and screened for the presence of pActin-OFP22 by kanamycin and rifampicin ^s -agrobacterium of 3Flag vector;

(2)pActin-OFP22 ^s -3Flag vector transformation of rice callus and obtaining rice seedlings regenerated: with pActin-OFP22 ^s EHA105 of the 3Flag vector infects rice callus, and after co-culturing for 3 days in a 28 ℃ culture room and washing out agrobacterium with a liquid medium, the rice callus is placed on a screening medium containing a proper antibiotic for culturing; after two rounds of culture, resistant calli can be obtained, and the resistant calli are transferredThe differentiation culture medium is used for carrying out differentiation culture to obtain seedlings, and the differentiated seedlings are transferred to a rooting culture medium for culture and transplanted after hardening.

Molecular detection of transgenic plants: according to OsOFP22 ^s CDS sequence of gene coding region and pActin-OFP22 ^s The 3Flag vector sequence creates forward and reverse primers, respectively, which are used to identify if the over-expression material was created successfully. The primer sequences were as follows:

sequence name	Sequence(s)	Sequence numbering
			pActin-F	5-TGCTGCTTCGTCAGGCTTAG-3	SEQ ID NO.5
OFP22s-OE-cs-R	5-CGTTGGGCATCATGTCGG-3	SEQ ID NO.6

The PCR product was 320bp.

Fluorescent reagent quantitative RT-PCR analysis: and (3) taking the extracted and stored rice leaf RNA, and carrying out reverse transcription to obtain the first strand cDNA. And (3) carrying out reagent quantitative PCR analysis, wherein the rice gene Actin01 is an endogenous reference gene, and detecting whether the expression quantity is improved. The primer sequences were as follows:

sequence name	Sequence(s)	Sequence numbering
			OFP22 ^s -qRT-F	5-GACATGATGCCCAACGC-3	SEQ ID NO.7
OFP22 ^s -qRT-R	5-AACCAAATCTCCCAACCCT-3	SEQ ID NO.8

。

The invention has the following beneficial effects: (1) OsOFP22 in the present invention ^s The protein amino acid and OFP22 protein are less than 88 amino acids at N terminal, and the expression analysis result shows that the identified short transcript OsOFP22 ^s The expression quantity in the rice seeds is far higher than that of OFP22 long transcripts, and the results of the three-generation transcriptome sequencing analysis of the rice development seeds show that OsOFP22 ^s Transcripts are the most predominant form of presence in rice seeds.

(2) Rice OsOFP22 in the invention ^s The gene has high expression in rice seeds, the protein product of the gene can directly interact with GS9, and more importantly, osOFP22 ^s After over-expression, the rice grain shape, thousand grain weight, amylose and other characters can be synergistically improved, and the rice has no obvious negative effect on other main agronomic characters, thus showing good breeding application value.

(3) Rice OsOFP22 is used in the invention ^s Construction of Gene sequence recombinant vector overexpressable in Rice, rice OsOFP22 ^s After the gene is over-expressed, the grain length and thousand grain weight of the rice are both improved compared with those of a control, and the grain width, thousand grain weight and physicochemical quality of the rice are obviously affected，OsOFP22 ^s The over-expressed rice has softer taste, improves the steaming quality of rice to a certain extent, and has important significance in improving the yield and quality of rice.

Drawings

FIG. 1 is GS9 and OsOFP22 ^s And (5) yeast double hybrid interaction verification of the protein.

FIG. 2 shows the in vivo luciferase complementation assay of tobacco to verify GS9 and OsOFP22 ^s Is described in (a) and (b) interact with each other.

FIG. 3 is an OsOFP22 ^s And OFP22 in seed.

FIG. 4 is a flower 11 and OsOFP22 ^s The phenotype of the mature seed grain of the over-expression material is that the top is middle flower 11 and the lower three rows are OsOFP22 respectively ^s Three different homologs of the over-expressed material.

FIG. 5 wild type flower 11 and OsOFP22 ^s Statistical comparison of the grain types of the overexpressed material rice.

FIG. 6 is a wild type flower 11 and OsOFP22 ^s Statistical comparison of thousand grain weight of rice over-expressed material.

FIG. 7 is a wild type flower 11 and OsOFP22 ^s Plant type comparison of the overexpressed material.

FIG. 8 is a wild type flower 11 and OsOFP22 ^s Comparison of rice amylose content (AAC) of the overexpressing material.

Detailed Description

The following examples are provided to illustrate the invention, but not to limit the scope of the invention, for the understanding of the invention.

The experimental procedures, which are not specified in the following examples, were carried out according to conventional procedures, using materials and being commercially available.

Example 1

GS9 and OsOFP22 ^s Verification of interaction

The interaction between proteins is the basis of cell vital activity, and is also an important component of the biochemical reaction network of cells, and has important significance for regulating and controlling cell signals. GS9 is an important granulocyte gene, in order to verify GS9 and OsOFP22 ^s ProteinsInteraction and further study of OsOFP22 ^s Specific protein segment of protein interaction with GS9 we will OsOFP22 ^s Full-length and N-and C-terminal truncated gene fragments are respectively constructed on pGADT7 vectors to form pGADT7-OsOFP22 ^s 、pGADT7-OsOFP22 ^s N、pGADT7-OsOFP22 ^s C multiple constructs (FIG. 1). The primer sequences were as follows:

sequence name	Sequence(s)	Sequence numbering
			OFP22 ^s -F	5-CGGAATTCATGGGCCGGCGGAAGTTCAG-3	SEQ ID NO.9
OFP22 ^s -R	5-CGGGATCCTCATGGCATGACGCCACAGG-3	SEQ ID NO.10
			OFP22 ^s -N-F	5-CGGAATTCATGGGCCGGCGGAAGTTCAG-3	SEQ ID NO.11
OFP22 ^s -N-R	5-CGGGATCCTCAGCTGTGCGCGAGCGGCG-3	SEQ ID NO.12
			OFP22 ^s -C-F	5-CGGAATTCATGTTCGCGGTGGTGAAGAC-3	SEQ ID NO.13
OFP22 ^s -C-R	5-CGGGATCCTCATGGCATGACGCCACAGG-3	SEQ ID NO.14
			GS9C1-F	5-GGAATTCCATATGCAGAGCAGCAGCAAGCG-3	SEQ ID NO.15
GS9C1-R	5-CGGAATTCCTAGCCTCTGGTTCGTATG-3	SEQ ID NO.16

。

After co-transformation of the above vector and pGBKT7-GS9C1 into Saccharomyces AH109, it was screened and identified on the corresponding auxotroph medium. The results showed full-length OsOFP22 ^s The protein interacts with GS9C1, truncated OsOFP22 ^s Only OsOFP22 ^s C interacts with GS9C1 (FIG. 1), illustrating OsOFP22 ^s Is the critical region that mediates its interaction with GS 9.

To further verify GS9 and OsOFP22 ^s Whether there was interaction in plants or not, we performed a luciferase complementation assay (Split Firefly Luciferase Comp lamentation Assay) validation. Full-length OsOFP22 ^s And GS9C1 were cloned into JW771 (35S-NLuc) and JW772 (35S-CLuc) vectors, respectively, with the primer sequences shown below:

。

the above-constructed correct vectors were transformed into Agrobacterium GV3101, respectively, inShake-cultured overnight in LB medium (5 mL) containing kanamycin. The appropriate combination is set, and the mixture is injected into tobacco epidermal cells, and after 48 hours, the activity of luciferase is detected by using a plant living molecule imaging system (CCD imaging system), so as to verify interaction. The results are shown in FIG. 2. Empty vector combinations NLuc and CLuc, empty vector and OsOFP22 ^s Or GS9C1 combinations, which contained only OsOFP22, were free of fluorescent signals ^s And GS9C 1. This experiment demonstrates GS9 and OsOFP22 ^s Also has interactions in plant cells.

Example 2

OsOFP22 ^s Gene overexpression recombinant vector pActin-OFP22 ^s Creation of-3 Flag

First strand cDNA was synthesized using wild type Japanese sunny RNA as a template, and OsOFP22 was used ^s The oligonucleotides at the 5 'and 3' ends of the gene coding sequence are used as PCR primers, and the sequences of the primers are as follows:

。

The plant binary expression vector pActin-3Flag (see www.cambia.org for details) was digested with Sam I and Xba I, and the digested vector was digested with homologous recombination enzymes to obtain a vector containing OsOFP22 ^s Ligation of PCR amplified products of full-length cDNA of the gene; obtaining the overexpression recombinant vector pActin-OFP22 ^s -3Flag; DH 5. Alpha. E.coli competent cells were transformed, screened by kanamycin on LB plate medium and verified by sequencing. Wherein the sequencing primer sequences are as follows:

sequence name	Sequence(s)	Sequence numbering
			p1300-3Flag-F	5-ATGGGGCTCTCGGATGTAGA-3	SEQ ID NO.21
p1300-3Flag-R	5-CGATCATAGGCGTCTCGCAT-3	SEQ ID NO.22

。

Example 3

Rice OsOFP22 ^s Method for applying gene in aspect of improving grain length, thousand grain weight and amylose content of rice

(2)pActin-OFP22 ^s -3Flag vector transformation of rice callus and obtaining rice seedlings regenerated: firstly, removing shells of mature Zhonghua 11 seeds, soaking and sterilizing the seeds for 1-2 hours by using a 2% sodium hypochlorite solution, continuously oscillating and washing the seeds during the sterilization period, then washing the seeds by using sterilized distilled water for 3-5 times, peeling young embryos on a sterile ultra-clean workbench by using pointed tweezers and a scalpel, transferring the embryos to an induction culture medium, and using the grown callus for subsequent genetic transformation. The single colony of the agrobacterium after colony-picking PCR verification is inoculated into 4mL of LB liquid medium containing 50mg/L kanamycin, and cultured at 28 ℃ and 250rpm overnight; transferring the culture medium into 50mL of the same culture medium according to the inoculum size of 1% in the next day, and performing expansion culture for 6-8h to logarithmic phase under the same conditions; centrifuging at 4000rpm and 4 ℃ for 5min, collecting thalli, then suspending in 10mL AAM liquid culture medium containing 100-400 mu mol/L acetosyringone, and subsequently infecting the peeled rice callus for 20min; after infection, pouring out the bacterial liquid, sucking the residual bacterial liquid by using sterile filter paper, transferring the callus into N ₆ D ₂ Culturing on C medium at 28deg.C in dark for 3 days, transferring the callus to N containing 600mg/L cephalosporin and 25mg/L hygromycin after 3 days ₆ D ₂ S ₁ Performing first-round screening culture on the culture medium; two weeks later transfer to new N containing 300mg/L cephalosporin and 50mg/L hygromycin ₆ D ₂ S ₂ Performing a second round of screening culture on the culture medium; transferring the resistant callus with strong activity to differentiation medium for differentiation culture, transferring the differentiated plantlet to 1/2MS ₀ Culturing on rooting culture medium, hardening seedling, and transplanting to transgenic rice test field.

(3) Molecular detection of transgenic plants: extracting target genome DNA in transgenic rice leaves to be detected according to OsOFP22 ^s CDS sequence of gene coding region and pActin-OFP22 ^s The 3Flag vector sequence creates forward and reverse primers, respectively, which are used to identify if the over-expression material was created successfully. The primer sequences were as follows:

。

And detecting whether the expression vector is successfully transferred into the rice or not through PCR amplification and gel electrophoresis. The size of the electrophoresis band of the PCR product is 320bp, if the plant with the same size as the target fragment is amplified, the plant is positive, otherwise, the plant is negative; on the other hand, screening positive transgenic plants were further confirmed by detecting hygromycin resistance of rice leaves ex vivo. Hygromycin detection primers were as follows:

sequence name	Sequence(s)	Sequence numbering
			Hyg-F	5-GCTTCTGCGGGCGATTTGTGT-3	SEQ ID NO.23
Hyg-R	5-GGTCGCGGAGGCTATGGATGC-3	SEQ ID NO.24

。

Example 4

Investigation of OsOFP22 gene overexpression in improving grain length and thousand grain weight of rice

OsOFP22 ^s The gene has high application value in the aspects of rice grain length and thousand grain weight after over-expression. After over-expression, the grain length is obviously improved compared with the control, and is shown in a specific figure 4, and a specific data statistical comparison is shown in a figure 5. Thousand kernel weight was significantly increased compared to the wild-type control, see in particular fig. 6. The gene and the genetic engineering technical means provided by the invention can obviously improve the rice yield on the premise of not affecting other agronomic traits of rice (figure 7), and have great application value.

Example 5

OsOFP22 ^s Gene overexpression obviously reduces the content of rice amylose

Analysis was carried out on the amylose content (AAC) which is the most important factor affecting the cooking taste quality (ECQ) of rice, and the results show that OsOFP22 ^s The overexpression of (2) significantly reduced AAC in rice (fig. 8). The results indicate OsOFP22 ^s The taste of the over-expressed rice is softer and glutinous, and the cooking quality of the rice is improved to a certain extent.

The foregoing description is only a preferred embodiment of the present invention, but is not intended to limit the present invention, and one skilled in the art may make possible variations and modifications to the present invention or modify equivalent embodiments with the technical content disclosed above without departing from the scope of the technical solution of the present invention, and any simple modifications, equivalent variations and modifications made to the above embodiments according to the technical substance of the present invention fall within the scope of the technical solution of the present invention.

Sequence listing

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<211> 30

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 15

ggaattccat atgcagagca gcagcaagcg 30

<210> 16

<211> 27

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 16

cggaattcct agcctctggt tcgtatg 27

<210> 17

<211> 39

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 17

acgggggacg agctcggtac catgggccgg cggaagttc 39

<210> 18

<211> 39

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 18

cgcgtacgag atctggtcga ctggcatgac gccacaggc 39

<210> 19

<211> 38

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 19

tacgcgtccc ggggcggtac ccagagcagc agcaagcg 38

<210> 20

<211> 39

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 20

cgaaagctct gcaggtcgac ctagcctctg gttcgtatg 39

<210> 21

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 21

atggggctct cggatgtaga 20

<210> 22

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 22

cgatcatagg cgtctcgcat 20

Claims

1. Overexpression of Rice OsOFP22 ^s Application of gene in aspects of improving grain length, thousand grain weight and reducing amylose content of rice, osOFP22 ^s The coding region sequence of the gene is shown as SEQ ID NO. 1.

2. The use according to claim 1, said OsOFP22 ^s The coded amino acid sequence is shown as SEQ ID NO. 2.

3. Root of Chinese characterThe use according to claim 1 or 2, characterized in that the method of use is as follows: with said rice OsOFP22 ^s The gene sequence constructs a recombinant vector capable of being over-expressed in rice, converts agrobacterium, transfers the vector into rice callus by using an agrobacterium-mediated rice genetic transformation method, and generates rice plants after resistance screening and tissue culture.