Rice gene osglw3 and its mutants and application

Abstract

The application discloses a rice gene OsGLW3 and a mutant and application thereof. The application clones a new dominant gene OsGLW3 for controlling rice grain type, plant type and ear type, the nucleotide sequence and the protein sequence coded by the gene are shown as SEQ ID NO: 1 and 3 respectively. In the mutant RE10498, the coding region 3119T of the gene is mutated into A, leading to early termination of the translated protein, and producing elongated grains. OsGLW3 codes an unknown expression protein, and the function of the gene is verified through complementary, knockout and overexpression experiments, and OsGLW3 is a functional gain type. RE The research on the molecular mechanism of OsGLW3 gene to rice grain type formation is of great significance, and the application provides excellent gene resources for rice variety improvement and hybrid breeding, and greatly accelerates the breeding process.

Description

Technical Field

[0001] This invention relates to the fields of genetic engineering and plant genetic breeding, and more specifically, to the rice gene OsGLW3 and its mutants and applications. Background Technology

[0002] Rice (Oryza sativa L.) serves as a fundamental food source for human survival, providing energy and nutrition to billions of people worldwide, and is a key element in ensuring food security and maintaining social stability.

[0003] Grain shape is closely related to the thousand-grain weight of rice. Generally speaking, longer and wider grains tend to have greater volume and weight. When the number of panicles and grains per panicle are relatively stable, appropriately increasing grain shape-related parameters can significantly improve the yield potential of rice. For example, under ideal cultivation conditions, some large-grain rice varieties can have a thousand-grain weight that is 20%-30% higher than ordinary varieties, thus increasing yield. Throughout the past few decades of rice breeding, breeders have continuously explored methods to improve yield by modifying grain shape. From traditional hybridization breeding to modern molecular breeding techniques, grain shape has been considered one of the important selection indicators.

[0004] Furthermore, rice grain shape significantly impacts the appearance, processing, and eating quality of rice. In terms of appearance, long, slender grains with low chalkiness are more popular in the market, often considered a key characteristic of high-quality rice by consumers. Rice with uniform grain shape is easier to husk and mill during processing, reducing broken rice and increasing milling yield, thus improving processing quality. Regarding eating quality, grain shape affects the cooking characteristics and texture of rice. Long-grain rice typically has better elasticity and extensibility after cooking, resulting in softer and more palatable rice, suitable for various dishes such as fried rice; while short-grain rice is relatively stickier, better suited for specific dishes like sushi. Different regions and consumer groups have varying demands for rice quality; therefore, developing rice varieties with different grain shapes to meet diverse market needs is particularly important.

[0005] Numerous studies have shown that grain shape is regulated by multiple genes and is a complex quantitative trait. Many rice grain shape genes have been cloned. For example, GS3 was the first major gene to be cloned and studied extensively, controlling rice grain length; its different allelic variations can significantly affect grain length (Fan et al., 2006; Mao et al., 2010; Li et al., 2016). GW2 and GW5 play important regulatory roles in grain width (Liu et al., 2017; Song et al., 2007), determining grain width development by influencing processes such as cell division and cell enlargement. GL7 and GW8 can alter rice quality while regulating grain shape (Wang et al., 2012; Yuexing et al., 2015).

[0006] All cloned grain shape genes are recessive or semi-dominant mutations. To date, there are no reports of dominant slender grain superior mutant genes being used in hybrid rice breeding. Therefore, creating dominant slender grain germplasm and cloning related genes, while simultaneously elucidating their genetic mechanisms, can provide specific germplasm resources for improving the quality of hybrid rice. Summary of the Invention

[0007] The purpose of this invention is to provide the rice gene OsGLW3 and its applications.

[0008] Another object of the present invention is to provide a mutant of the rice gene OsGLW3, OsGLW3. RE And its applications.

[0009] To achieve the objectives of this invention, in a first aspect, this invention provides a rice gene OsGLW3, which is a gene encoding either protein (A) or (B) as follows:

[0010] (A) A protein consisting of the amino acid sequence shown in SEQ ID NO: 3; or

[0011] (B) A protein derived from (A) with the sequence shown in SEQ ID NO: 3 replaced, deleted or added with one or more amino acids and having the same function.

[0012] The nucleotide sequence of the gene OsGLW3 is as follows:

[0013] I) The nucleotide sequence shown in SEQ ID NO: 1;

[0014] II) A nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 1 that has been substituted, deleted and / or added with one or more nucleotides and expresses a protein with the same function;

[0015] III) A nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO: 1 under stringent conditions and expresses a protein with the same function, wherein the stringent conditions are hybridization at 65°C in 0.1×SSPE containing 0.1% SDS or 0.1×SSC containing 0.1% SDS, followed by washing the membrane with the same solution; or

[0016] A nucleotide sequence that has more than 90% homology with the nucleotide sequences of I), II) or III) and expresses the same functional protein.

[0017] Its cDNA sequence is:

[0018] i) The nucleotide sequence shown in SEQ ID NO: 2;

[0019] ii) A nucleotide sequence of the nucleotide sequence shown in SEQ ID NO: 2 that has been substituted, deleted and / or added with one or more nucleotides and expresses a protein with the same function;

[0020] iii) A nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO: 2 under stringent conditions and expresses a protein with the same function, wherein the stringent conditions are hybridization at 65°C in 0.1×SSPE containing 0.1% SDS or 0.1×SSC containing 0.1% SDS, followed by washing the membrane with the same solution; or

[0021] iv) Nucleotide sequences that have more than 90% homology with the nucleotide sequences of i), ii) or iii) and express the same functional protein.

[0022] Secondly, the present invention provides biological materials containing the gene OsGLW3, including but not limited to recombinant DNA, expression cassettes, transposons, plasmid vectors, viral vectors, engineered bacteria, or non-renewable plant parts.

[0023] Thirdly, the present invention provides any of the following applications of the gene OsGLW3 or biological materials containing the gene:

[0024] (1) Regulate plant grain type, plant type and spike type;

[0025] (2) Used for plant variety improvement;

[0026] (3) Used to prepare transgenic plants.

[0027] In this invention, the rice grain type includes grain length, grain width, and grain length-to-width ratio; the plant type includes plant height; and the panicle type includes panicle length.

[0028] In this invention, the plant is a grass, preferably rice.

[0029] Fourthly, the present invention provides a rice OsGLW3 gene mutant, OsGLW3. RE It is a gene that encodes either protein (a) or (b) as follows:

[0030] (a) A protein consisting of the amino acid sequence shown in SEQ ID NO: 4; or

[0031] (b) Proteins derived from (a) with the sequence shown in SEQ ID NO: 4 substituted, deleted or added with one or more amino acids and having the same function.

[0032] Gene mutant OsGLW3 RE The mutation at position 3119 of the coding region of the rice OsGLW3 gene, where a T-to-A base was changed, caused premature termination of protein translation.

[0033] Fifthly, the present invention provides a gene mutant OsGLW3. RE Biological materials, including but not limited to recombinant DNA, expression cassettes, transposons, plasmid vectors, viral vectors, engineered bacteria, or non-renewable plant parts.

[0034] Sixthly, the present invention provides the gene mutant OsGLW3. RE Or any of the following applications of biological materials containing the said gene mutant:

[0035] (1) Regulate plant grain type, plant type and spike type;

[0036] (2) Used for plant variety improvement;

[0037] (3) Used to prepare transgenic plants.

[0038] Seventhly, the present invention provides a method for increasing the length of rice grains, narrowing the grain width, increasing the length-to-width ratio, reducing grain weight, increasing plant height, making the flag leaf thinner and longer, and increasing the panicle length. The method includes: using genetic engineering techniques to overexpress the mutant gene OsGLW3 in rice. RE .

[0039] The overexpression method can be selected from the following 1) to 5), or any combination thereof:

[0040] 1) By importing a plasmid containing the gene;

[0041] 2) By increasing the copy number of the aforementioned genes on plant chromosomes;

[0042] 3) By altering the promoter sequence of the aforementioned genes on plant chromosomes;

[0043] 4) By operatively linking a strong promoter to the gene;

[0044] 5) By introducing enhancers.

[0045] Expression vectors carrying the target gene can be introduced into plant cells using conventional biotechnological methods such as Ti plasmids, plant virus vectors, direct DNA transformation, microinjection, and electroporation (Weissbach, 1998, Method for Plant Molecular Biology VIII, Academy Press, New York, pp. 411-463; Geiserson and Corey, 1998, Plant Molecular Biology, 2nd Edition).

[0046] Eighthly, the present invention provides the application of transgenic rice obtained according to the above method in plant breeding.

[0047] The breeding methods include, but are not limited to, transgenic, hybrid, backcross, self-crossing or asexual reproduction.

[0048] By employing the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0049] (I) This invention provides the rice gene OsGLW3. This invention clones a novel rice gene, OsGLW3, that controls grain shape. The coding region of this gene has a 3119T mutation to A, resulting in gain-of-function, specifically increased grain length and decreased grain width and weight. The OsGLW3 gene is of great significance for studying the molecular mechanism of rice grain shape formation. This invention provides a new gene resource for rice grain shape improvement breeding, greatly advancing the rice breeding process.

[0050] (ii) By crossing mutants carrying OsGCW3 with sterile lines, the grain shape and appearance quality of hybrid rice can be improved.

[0051] (III) By constructing OsGLW3 RE Transgenic plants of wild-type Zhonghui 8015, after overexpression vector, exhibit increased grain length. This can be used to improve rice grain length and adapt it to market demands. Attached Figure Description

[0052] Figure 1 Mature kernels of ZH8015 and RE10498 are shown in the preferred embodiment of this invention. The scale bar is 5 mm. CF: Kernel width (C), kernel length (D), length-to-width ratio (E), and thousand-kernel weight (F) of ZH8015 and RE10498. Data: Mean ± Standard Deviation (n = 20). P-values ​​were obtained using a two-tailed t-test.

[0053] Figure 2This is a comparison of agronomic traits between ZH8015 and RE10498 in a preferred embodiment of the present invention. A, B: Comparison of plant type and panicle type of ZH8015 and RE10498, scale bar: A, 10cm; B, 5cm. CM: Panicle length (C), number of primary branches (D), number of secondary branches (E), flag leaf length (F), flag leaf width (G), plant height (H), number of effective panicles (I), total number of grains per plant (J), seed setting rate (K), number of filled grains per panicle (L), and yield per plant (M) of ZH8015 and RE10498. Data: mean ± standard deviation (n = 20). P-value was obtained by two-tailed t-test.

[0054] Figure 3 This is a cytological observation of glumes ZH8015 and RE10498 in a preferred embodiment of the present invention. A: Phenotype, scale bar 5mm; B: Cross-sectional view of glumes, scale bar 50μm; C: Enlarged view of the area within the red box in Figure B, scale bar 100μm; DE: Total length (D) and cell width (E) of the inner thin-walled cells in the cross-section; F: Scanning electron microscopy of the outer surface of the glumes, scale bar 400μm; GH: Number of cells per unit area (G), cell length (H), and cell width (I). Data: Mean ± Standard Deviation. P-values ​​were obtained using a two-tailed t-test.

[0055] Figure 4 This is a distribution chart of the number of times the seed quality data of 314 individual plants in the F2 generation of the present invention were tested. A: 1000-grain weight; B: length-to-width ratio; C: grain length; D: grain width.

[0056] Figure 5 This is a distribution map of Δ_All-index on chromosomes in MutMap in a preferred embodiment of the present invention.

[0057] Figure 6 This is a functional verification of OsGLW3 in a preferred embodiment of the present invention. COM represents the genomic sequence of wild-type OsGLW3 transformed into the mutant, and OE represents OsGLW3 overexpressed in the mutant within the wild type. RE The coding sequence, KO-1 is the OsGLW3 knockout in the wild type. ZH KO-2 is a mutant in which OsGLW3 is knocked out. RE A. Grain phenotype; B. Grain length-to-width ratio; C. Grain length; D. Grain width. Data: mean ± standard deviation (n > 6), Duncan multiple comparisons.

[0058] Figure 7This section analyzes the expression pattern of OsGLW3 in a preferred embodiment of the present invention. A: Expression levels of the OsGLW3 gene in different locations in ZH8015 and RE10498. Root: root; Node: node; Leaf: leaf; P0-1cm, P1-3cm, P3-5cm, P5-7cm, P7-10cm, P10-13cm, P13-17cm represent different spike lengths. GUS staining of transgenic plants: root (B), stem (C), leaf (D), leaf sheath (E), spike (F), and glumes (G, H). The scale bar for B and F is 2cm, and the scale bars for G and H are 2mm. Data: mean ± standard deviation. P-values ​​were obtained using a two-tailed t-test. Detailed Implementation

[0059] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the examples are conducted under conventional experimental conditions, such as those described in Sambrook et al., Molecular Cloning: a Laboratory Manual (Sambrook J & Russell DW, 2001), or as recommended by the manufacturer's instructions.

[0060] The rice material used in this invention, Zhonghui 8015 (ZH8015), is a strong three-line restorer line bred by crossing the restorer line Zhonghui 218 as the female parent and Zhonghui 8006 as the male parent. In this invention, ZH8015 was mutagenized with ethyl methanesulfonate, and the slender grain mutant RE10498 was screened from its mutant library. RE10498 and ZH8015 were crossed, and the grains of their BC1F2 were tested. Grain width and thousand-grain weight showed a continuous normal distribution, while grain length and length-to-width ratio showed varying degrees of segregation. Chi-square tests on grain length showed a segregation ratio of 1:3, indicating that grain length is controlled by a pair of dominant genes. The candidate gene LOC_Os03g21270 was obtained using the MutMap method. In its coding region, position 3119 is changed from thymine (T) to adenine (A), resulting in a leucine mutation into a stop codon, causing premature termination of translation. We hypothesized that LOC_Os03g21270 was the target gene and named it OsGLW3. We then verified the gene's function through transgenic analysis.

[0061] Spatiotemporal expression and GUS staining of OsGLW3 gene: RNA was extracted from various rice tissue samples, and spatiotemporal expression analysis of OsGLW3 was performed; the pCAMBLA1305-GUS vector was constructed to verify the spatiotemporal expression results.

[0062] Example 1: Gene Localization

[0063] We identified a novel rice grain type mutant, RE10498, which, compared to the wild type, exhibited increased grain length, decreased grain width, and decreased grain weight. Scanning electron microscopy of the glumes revealed a decrease in the width of the epidermal cells and a reduction in the number of transverse cells, consistent with the grain type changes. Genetic analysis indicated that grain length was controlled by a single dominant gene pair. Using the MutMap method, we obtained the candidate gene LOC_Os03g21270, whose coding region showed a single base change (T>A) at position 3119, resulting in a leucine mutation to a stop codon, causing premature translation termination. Subsequent transgene complementation, knockout, and overexpression experiments confirmed that LOC_Os03g21270 was the cause of the mutant phenotypic changes, and it was named OsGLW3. Among these, after constructing a CRISPR / Cas9 gene knockout vector and transforming it into ZH8015, the transgenic plants showed no significant changes in grain size. However, after transforming into the mutant RE10498, the transgenic plants had shorter grain length, increased grain width, and increased grain weight, similar to the wild type. Gene function was verified using a complementation experiment. Transforming the mutant RE10498 with the pOsGLW3::OsGLW3 complementation vector resulted in grain length, width, and aspect ratio that were between those of the wild type and the mutant. Gene function was further verified using overexpression with pUbi::OsGLW3. RE Transformation of wild-type ZH8015 with the overexpression vector resulted in transgenic plants with longer grains, narrower grains, and lower grain weight, similar to the mutant. OsGLW3 encodes an expression protein of unknown function and is a novel, previously unreported gene in rice.

[0064] 1. Particle shape phenotype

[0065] We used EMS to mutagenesis of ZH8015 and screened for the slender-grained mutant RE10498 in the mutant library. Phenotypic analysis revealed that, compared to the wild-type ZH8015, the mutant RE10498 produced slender and lighter grains. Figure 1 The mutant RE10498 showed a 5.82% increase in grain length, a 7.53% decrease in grain width, a 14.73% increase in length-to-width ratio, and a 5.93% decrease in thousand-grain weight, all of which were highly significant.

[0066] 2. Agronomic traits and yield traits

[0067] We examined the correlation between plant type and yield traits between ZH8015 and RE10498. The results showed that the mutant RE10498 had a significantly increased plant height compared to the wild type, and its flag leaf was also slender, resulting in a slightly looser plant type. Figure 2 (A and FH). In terms of panicle type, RE10498 showed a highly significant increase in panicle length, number of primary branches, and number of secondary branches. Figure 2(B and CE). Compared with the wild type, the mutant showed a highly significant increase in the total number of grains per plant, a significantly decreased seed setting rate, and no significant difference in the number of effective tillers. Since the mutant had a 7.44% increase in the number of grains per ear, it compensated for the decrease in grain weight, ultimately resulting in no significant difference in yield per plant. Figure 2 ,IM).

[0068] 3. Cytological observation

[0069] Rice grain size is typically regulated by the number and size of glume cells. To investigate how mutants alter rice grain size, we observed the cells in the glume cross-sections of ZH8015 and RE10498 before flowering. The mutant RE10498 showed a significant decrease in both the total length and cell width of the inner parenchyma cells. Figure 3 Scanning electron microscopy (SEM) of the outer surface cells of the glumes revealed that the mutant had an increased cell number and a significantly decreased cell width compared to the wild type, while the cell length showed no significant change. Figure 3 The above results indicate that OsGLW3 makes the grains slender by regulating longitudinal glume cell proliferation and reducing transverse cell width.

[0070] 4. Genetic analysis and gene mapping

[0071] RE10498 and ZH8015 were crossed, and the grains of their BC1F2 strain were tested. Grain width and thousand-grain weight showed a continuous normal distribution, while grain length and length-width ratio showed varying degrees of segregation. Figure 4 Chi-square test on grain length showed that the grain length conformed to a segregation ratio of 1:3, indicating that grain length is controlled by a pair of dominant genes (Table 1).

[0072] Table 1. Chi-square test of kernel length in BC1F2 generation

[0073]

[0074] Note: GL-L and GL-H represent grain lengths of 9.620–10.370 mm and 10.371–11.15 mm, respectively.

[0075] Thirty plants each with long and short grains were selected from BC1F2 to construct a mixed pool. Four candidate sites were obtained using the MutMap method. Figure 5(See Table 2). In the coding region of OsR498G0305879600.01 (LOC_Os03g21270), the thymine (T) is converted to adenine (A) at position 3119, resulting in a leucine mutation that becomes a stop codon, causing premature termination of translation. The remaining three candidate sites are all located upstream or downstream of the gene; therefore, we speculate that LOC_Os03g21270 is the target gene and name it OsGLW3.

[0076] Table 2 Candidate Gene List

[0077]

[0078] Example 2: Function of gene OsGLW3

[0079] In this embodiment, a complementation experiment was used to transform pOsGLW3::OsGLW3 into the mutant RE10498. The length and width of the transgenic plants were between those of the wild type and the mutant. Furthermore, a gene knockout experiment was conducted to verify gene function. By constructing a CRISPR / Cas9 gene knockout vector and transforming it into the wild-type ZH8015, no significant changes were observed in the grain size of the transgenic plants. However, the transgenic plants transformed into the mutant RE10498 showed decreased grain length but increased grain width and weight. Furthermore, we used the OsGLW3 from the mutant... RE Using the encoded sequence as a template, construct pUBi::OsGLW3 RE When the overexpression vector was transformed into wild-type ZH8015, the transgenic plants showed increased grain length, decreased grain width, and decreased grain weight. Details are as follows:

[0080] 1. Complementarity Verification

[0081] To verify whether the variation in the candidate gene OsGLW3 led to changes in the grain size and plant type of the mutant, a transgenic complementation experiment was first conducted. The full-length 10.543 kb OsGLW3 genome was integrated into a complementation vector (pCAMBIA1300, purchased from CAMBIA). Using Agrobacterium-mediated transformation, the constructed complementation vector pOsGLW3::OsGLW3 was introduced into the RE10498 mutant. Positive transgene verification in the T0 generation was performed using a combination of hygromycin-specific primer PCR amplification and mutation site sequencing. Species testing revealed that the grain length and width of the T1 generation complementary plants were between those of the wild type and the mutant. Figure 6 ).

[0082] 2. Knockout Verification

[0083] The knockout vector we used was the pCas9-sgRNA vector (MIAO et al., 2013). This vector has prokaryotic resistance to spectinomycin and eukaryotic resistance to hygromycin. AarI was used as the restriction site during vector construction. The target sequence of the gene was designed using an online website (http: / / cbi.hzau.edu.cn / cgi-bin / CRISPR2 / CRISPR). The high-scoring sequence N18 (5'-TGAAGCAGACCTCATGAG-3') was selected, and the corresponding adapter was added to form the F primer (5'-AGATGATCCGTGGCATGAAGCAGACCTCATGAGGTTTTAGAGCTATGC-3'). The F primer sequence was reverse complementary to form the R primer. Add 1 μL of F and R primers to a 10 μL system, anneal at 94℃ for 10 min, then anneal at 0.1℃ / s to 15℃, and hold at 15℃ for 10 min to complete annealing. Take 1 μL of the annealing product and recombinant it with the enzyme-digested pCas9-sgRNA vector plasmid, transform it into DH5α, and plate it on spectinomycin-resistant solid medium. After successful single-clone sequencing, the plasmid was sent to Wuhan Boyuan Biotechnology Co., Ltd., and transformed into ZH8015 background. Compared with ZH8015, there was no significant difference in seed length and length-to-width ratio in positive plants, but a slight difference in seed width. Figure 6 Subsequently, we transformed pCas9-sgRNA recombinant plasmids containing the same sgRNA into the RE10498 background. Compared with RE10498, the positive plants showed decreased grain length and increased grain width, exhibiting grains more similar to ZH8015. Figure 6 The above results indicate that RE10498 is a gain-of-function mutant.

[0084] 3. Overexpression verification

[0085] To further verify the presence of OsGLW3 in the mutant RE Regulation of grain shape. We amplified the OsGLW3 mutant sequence (SEQ ID NO: 5) using RE10498 cDNA as a template, and homologously recombinated it into the overexpression vector pCAMBIA1301 (purchased from CAMBIA, maize Ubiquitin promoter) to construct the fusion vector pUbi::OsGLW3. RE The vector was introduced into ZH8015 using Agrobacterium-mediated transformation (Wuhan Boyuan Biotechnology Co., Ltd.). Families with high expression levels were selected from T2-stable positive transgenic plants, and the results showed that OsGLW3 was overexpressed in the ZH8015 background. RE This will result in longer, narrower, and lighter kernels. Figure 6 ).

[0086] The above research results indicate that OsGLW3 REThis manifests as functional gains: increased grain length, decreased grain width, and decreased grain weight.

[0087] Example 3: Expression pattern analysis of gene OsGLW3

[0088] To investigate the spatial expression of the gene OsGLW3, we used quantitative real-time PCR to analyze its expression levels in different parts of the plant. The results showed that OsGLW3 was expressed in roots, stems, leaves, leaf sheaths, and panicles, but its expression level was higher in the panicle. Figure 7 (A). Furthermore, we validated this result using transgenic plants with β-glucuronidase (GUS) reporter gene expression initiated by the OsGLW3 promoter: GUS staining was detected in roots, stems, leaves, leaf sheaths, and spikes, with the deepest staining in the spikes. Figure 7 (BH). This indicates that OsGLW3 is constitutively expressed, and its expression level is highest in the spikelet.

[0089] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

[0090] References:

[0091] 1. FAN CC, Genetics, 112(6): 1164-1171. DOI: 10.1007 / s00122-006-0218-1.

[0092] 2、LIU J F,CHEN J,ZHENG X M,WU F Q,LIN Q B,HENG Y Q,TIAN P,CHENG Z J,YU X W,ZHOU KN,ZHANG X,GUO X P,WANG J L,WANG H Y and WAN J M,2017.GW5 acts inthe brassinosteroid signalling pathway to regulate grain width and weight inrice.Nature Plants,3:17043.DOI:10.1038 / nplants.2017.43.

[0093] 3、MAO H L,SUN S Y,YAO J L,WANG C R,YU S B,XU C G,LI X H and ZHANG QF,2010.Linking differential domain functions of the GS3 protein to naturalvariation of grain size in rice.Proc Natl Acad Sci U S A,107(45):19579-19584.DOI:10.1073 / pnas.1014419107.

[0094] 4、MIAO J,Guo D S,ZHANG J Z,HUANG Q P,QIN G J,ZHANG X,WAN J M,GU H Yand QU L J,2013.Targeted mutagenesis in rice using CRISPR-Cas system.CellResearch,3;23(10):1233-1236.DOI:10.1038 / cr.2013.123.

[0095] 5、SONG X J,HUANG W,SHI M,ZHU M Z and LIN H X,2007.A QTL for ricegrain width and weight encodes a previously unknown RING-type E3 ubiquitinligase.Nature Genetics,39(5):623-630.DOI:10.1038 / ng2014.

[0096] 6. WANG SK, WU K, YUAN QB, LIU XY, LIU ZB, LIN XY, ZENG RZ, ZHU HT, DONG GJ, QIAN Q, ZHANG GQ, FU XD, 2012. Control of grain size, shape and quality by OsSPL 16in rice.Nature Genetics, 44(8):950-954.DOI: 10.1038 / ng.2327.

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[0098] 8. Li Yang, Xu Xiaoyan, Yan Ming, Feng Fangjun, Ma Xiaosong and Mei Hanwei, 2016. Study on improvement of rice grain type using functional molecular markers of GS3 gene. Shanghai Journal of Agricultural Sciences (1): 1-5. DOI: 10.15955 / j.issn1000-3924.2016.01.01.

Claims

1. Rice genes OsGLW3 Or the application of the genetically modified biomaterials containing the aforementioned genes in regulating the reduction of grain length and the increase of grain width in rice, wherein, The gene OsGLW3 is a gene encoding the following protein (A): (A) a protein consisting of the amino acid sequence shown in SEQ ID NO:3; its nucleotide sequence is: I) the nucleotide sequence shown in SEQ ID NO:

1.

2. Rice OsGLW3 Gene mutants OsGLW3 RE Its characteristics are, It is the gene encoding the following protein (a): (a) A protein consisting of the amino acid sequence shown in SEQ ID NO:

4.

3. A biological material containing the gene mutant of claim 2, wherein the biological material is recombinant DNA, expression cassette, transposon, plasmid vector, viral vector, or engineered bacteria.

4. The following applications of the gene mutant of claim 2 or the biomaterial of claim 3: Regulating rice grains can increase grain length, narrow grain width, increase length-to-width ratio, decrease grain weight, increase plant height, make flag leaves thinner and longer, and increase panicle length.

5. A method for making rice grains longer, narrower, with a higher length-to-width ratio, lower in weight, taller, with thinner and longer flag leaves, and with a longer panicle, characterized in that: The method includes: using genetic engineering techniques to overexpress a gene mutant in rice. OsGLW3 RE Among them, gene mutants OsGLW3 RE As described in claim 2.

6. The method according to claim 5, characterized in that, The overexpression method is selected from the following 1) to 5), or any combination thereof: 1) By importing a plasmid containing the gene; 2) By increasing the copy number of the aforementioned genes on the rice chromosome; 3) By altering the promoter sequences of the aforementioned genes on the rice chromosome; 4) By operatively linking a strong promoter to the gene; 5) By introducing enhancers.

7. The application of the transgenic rice obtained according to the method of claim 5 or 6 in rice breeding.

8. The application according to claim 7, characterized in that, Breeding methods include transgenic, hybridization, backcrossing, self-pollination, or asexual reproduction.

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