CN109439672B - Application of gene ESP for regulating development of rice panicle - Google Patents

Abstract

The invention discloses an application of a gene ESP for regulating and controlling development of rice ears, belonging to the field of plant genetic engineering. The invention clones the gene ESP for regulating the development of rice ears by adopting a map-based cloning method, and the gene ESP has the nucleotide sequence shown as SEQ ID NO: 1; and the important function of the ESP gene in regulating the rice panicle type is proved through mutant phenotype analysis and complementary analysis. The cloning of the ESP gene not only enriches the molecular mechanism of the regulation of the rice panicle type, has important theoretical significance in disclosing the development mechanism of the rice panicle, but also provides a new clue for researching the evolution of the rice panicle type. Meanwhile, a new gene resource is provided for the genetic improvement of the rice panicle type, and a new material selection is provided for the molecular design breeding of rice. The method can cultivate the vertical dense-panicle rice variety by improving the expression of the ESP in the rice, has strong operability, and opens up a new way for widening the genetic basis of the existing vertical panicle type variety.

Description

Application of gene ESP for regulating development of rice panicle

Technical Field

The invention belongs to the field of plant genetic engineering, in particular relates to application of a gene ESP for regulating development of rice panicles, and specifically relates to application of a gene ESP for controlling rice panicle type separated cloning by using a map-based cloning strategy, biological function verification and genetic transformation of the gene and application of the gene ESP as a panicle development regulating gene in rice panicle type genetic improvement.

Background

Rice is an important food crop in China and even the world, and rice is taken as staple food by more than half of the population all over the world. The panicle length and the grain density of the rice are important factors influencing the yield, so that the elucidation of the genetic basis and the molecular control mechanism of panicle development is helpful for improving the panicle type and the yield of the rice.

The panicle type of the rice has important significance in natural evolution and artificial domestication research. In the domestication process from wild rice to cultivated rice, the panicle type of the rice is changed greatly, the wild rice is loose and has low density of panicle type, while the cultivated rice is relatively compact and has high density of panicle type. Therefore, the research on the genetic mechanism of the development of the panicle can provide strong evidence and clues for the evolutionary research of rice.

The epigenetic variation greatly enriches the phenotypic diversity of eukaryotes, and the epigenetic allele can provide a new genetic resource for crop breeding. At present, relatively few reports on important agronomic traits controlled by epigenetic inheritance exist in rice, and the case of applying epigenetic variation to rice breeding is more rare. Therefore, the cloning of the gene for controlling important agronomic traits of rice by epigenetic inheritance has important significance for genetic improvement and enrichment of variety genetic diversity of rice.

The spike type is an important agronomic trait of rice, is determined by a plurality of factors including spike length, primary and secondary stem number, glume flower/seed number, seed weight and the like, and has important significance for the modeling of the spike type of the rice and the improvement of the rice yield by clarifying a spike forming mechanism. In recent years, scientists have conducted extensive and intensive studies on the regulation of the panicle type of rice, and have made important progress (koyan, li courage, progress in the genetic regulation of the panicle type of rice. plant proceedings, 2017(01): 19-29). However, the current knowledge of the molecular regulation mechanism of spike formation is still very limited.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide the gene ESP for regulating and controlling the development of rice ears. The gene plays an important role in rice panicle type regulation, so that the cloning of the gene is beneficial to enriching the molecular mechanism of the rice panicle type regulation.

The invention screens a mutant esp (epidemic short panicle) from the offspring of the wild rice middle flower 11, the mutant rice spike is shortened, the grain density is increased, and the spike is upright. Genetic analysis shows that the mutation is incompletely dominant. Mutant candidate genes Os01g0356951 have been obtained by using a map-based cloning strategy, and no research on the functions of the genes exists at present. Therefore, the ESP gene of the present invention is a novel gene for regulating the development and panicle type of rice panicle.

The invention also aims to provide application of the gene ESP for regulating and controlling the development of rice ears.

The purpose of the invention is realized by the following technical scheme:

the invention provides an application of a gene ESP for regulating development of rice ears in regulating development and ear types of the rice ears.

Furthermore, the gene ESP for regulating the development of the rice panicle is applied to rice panicle type improvement breeding.

Furthermore, the gene ESP for regulating the development of the rice panicle is applied to culturing the vertical dense panicle rice variety.

The gene ESP for regulating rice spike development is the following nucleotide sequence A or B:

A. sequence listing SEQ ID NO: 1;

B. the analogue obtained by the above A through base insertion, deletion or substitution still has the function of regulating the development and the spike type of the rice.

The application of the recombinant expression vector, the transgenic cell line and the recombinant strain containing the gene ESP for regulating and controlling the development of the rice panicle also belongs to the protection scope of the invention.

The rice ESP mutant plant of the invention is slightly short, the spike length is obviously shortened, the spike shape is upright and the density of the planted grain is increased.

2) Gene expression analysis shows that the expression of ESP gene in mutant is obviously improved. However, sequencing analysis revealed that no DNA sequence change was observed in the ESP gene (including the promoter region and the 3' -end downstream region of the gene). Therefore, we speculate that epigenetic variation of the gene of interest may occur.

3) Based on the above presumption, the present applicant analyzed methylation modification variation by bisulfite sequencing. DNA methylation modification is closely related to gene expression regulation. Generally, methylation, if it occurs, tends to inhibit gene expression, while demethylation causes overexpression of the gene. The research of the invention shows that the ESP gene overexpression is caused by the methylation abnormality (demethylation) of the adjacent section at the downstream of the 3' end of the ESP gene.

4) As the mutant phenotype is not caused by ESP functional deletion, in order to further verify the function, an ESP overexpression vector pCUbi1390-ESP-OE driven by a Ubi (Ubiquitin) promoter of corn is constructed, a wild type rice middle flower 11 is transformed through mediation of agrobacterium (EHA105), and a transgenic positive plant is detected through PCR. The results show that the ESP overexpression transgenic plant has similar phenotype to the mutant. The function complementation test shows that the candidate gene ESP obtained by the invention is a target gene for controlling the rice panicle type.

5) The application of the rice ESP gene refers to the application of the gene in rice panicle type improvement breeding. The rice panicle type improved breeding mode comprises transgenic breeding and crossbreeding, and the method comprises the following steps: transgenic breeding, wherein a recombinant expression vector containing an ESP gene is introduced into plant cells, and the expression level of the ESP gene is changed to obtain transgenic rice with changed panicle type, for example, the rice with improved expression of the ESP gene can be used for breeding vertical dense panicle rice varieties. And (3) cross breeding, namely, using the transgenic rice with the ear type variation as a parent, and improving the ear type of the rice by a cross breeding method, namely, hybridizing the obtained transgenic rice with the changed ear type with a rice variety to be improved, and then breeding the rice variety with the improved ear type.

The recombinant expression vector containing the ESP gene can be constructed by the existing plant expression vector; the plant expression vector comprises a binary agrobacterium vector and a vector which can be used for plant microprojectile bombardment, such as pCAMBIA1300, pCAMBIA2300, pCUbi1390 and the like; the recombinant expression vector containing ESP gene can be introduced into host plant cell or tissue by various physical, chemical and biological methods such as gene gun, microinjection, protoplast mediation, pollen tube channel, agrobacterium mediation and the like; the host plant to be introduced is preferably rice.

When constructing a recombinant expression vector containing the ESP gene, any one of enhanced, constitutive, tissue-specific or inducible promoters, such as 35S promoter, Ubi promoter, etc., may be added before the transcription initiation nucleotide of the ESP gene.

In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used may be modified for use, including the addition of antibiotic resistance genes or other marker genes.

Compared with the prior art, the invention has the following advantages and effects:

1. the invention clones an epigenetic modification regulated rice panicle development gene ESP from rice by a map-based cloning method, and the mutant phenotype analysis and complementary analysis show that the obtained ESP gene has an important role in rice panicle type regulation, provides a new gene resource for rice panicle type genetic modification, and provides a new material selection for rice molecular design breeding.

2. The rice ears can be divided into three types, i.e., curved ears, semi-upright ears and upright ears. In the grouting and maturing period, the vertical spike shape can effectively reduce shading, is beneficial to improving the colony structure and the light receiving situation, and has an important effect on increasing the yield of the rice. The method can cultivate vertical dense-panicle rice varieties by improving the expression of the ESP in the rice, has strong operability, and opens up a new way for widening the genetic basis of the existing vertical panicle type varieties.

3. The invention proves the function of ESP in the regulation of the rice panicle type, enriches the molecular mechanism of the regulation of the rice panicle type, has important theoretical significance in disclosing the development mechanism of the rice panicle, and provides a new clue for researching the rice panicle type evolution.

Drawings

FIG. 1 shows the phenotypes of the esp mutant and wild type (japonica rice, mid-flower 11) in example 1; wherein, panels A and B are respectively plant type and panicle type of chimeras and mutant hybrids in wild type and mutant hybrids, and panels C, D and E are respectively plant phenotype, main stem and scion morphology at the panicle development stage of wild type and mutant homozygote; WT is a wild type, Epi-sp (+/-) is a mutant hybrid, Epi-normal is a spike of rice in which the chimera has a normal phenotype in the mutant hybrid, Chimeric is a chimera in the mutant hybrid, Epi-sp is a spike of rice in the mutant hybrid, and Epi-sp (-/-) is a mutant homozygote.

FIG. 2 shows the panicle and panicle traits of flower 11 in the esp mutant and wild type in example 2, wherein Panel A shows the panicle and grain type of the wild type and mutant hybrids, and FIGS. B, C and D show the panicle length, primary branch length and primary branch number of the wild type and mutant hybrids, respectively.

FIG. 3 is a map-based cloning and expression analysis of ESP genes in example 3, wherein, Panel A is a marker and candidate gene analysis for map-based cloning, Panel B is a relative expression analysis of 3 genes (Os01g0356900, Os01g0357100 and Os01g0357200) in the localization region shown in Panel A, Panel C is an expression of ESP gene (Os01g0356951) in wild type, mutant heterozygote and mutant homozygote, Panel D is a gene expression analysis of ESP gene in normal rice panicle (N, i.e., Epi-normal) and mutant phenotype (sp, i-sp) rice panicle of chimeric mutant, and Panel E is a gene expression analysis of ESP gene in the progeny of chimera with different phenotypes; 25sRNA is an internal reference gene for gene expression analysis.

FIG. 4 is DNA methylation and gene expression analysis of the ESP gene in example 4, wherein FIG. A and FIG. B show the difference in DNA methylation at the 3' -end of the ESP gene between the wild type and the mutant, and FIG. C shows the expression analysis of the ESP gene after flower 11 in the wild type was treated with the methylation inhibitor 5-aza-dC.

FIG. 5 shows the phenotype of ESP gene overexpressed transgenic plant in example 5, wherein panel A shows the phenotype of wild type and individual plant overexpressing ESP gene, and panel B shows the phenotype of wild type and rice ear overexpressing ESP gene; OE is an overexpression transgene phenotype.

Detailed Description

The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.

The test methods in the following examples, in which specific experimental conditions are not specified, are generally performed according to conventional experimental conditions or according to the experimental conditions recommended by the manufacturer. The materials, reagents and the like used are, unless otherwise specified, reagents and materials obtained from commercial sources.

In the embodiment, the pCUbi1390 vector is a commonly used overexpression vector, pEasy-T1 (Beijing holotype gold), Escherichia coli DH5 α and Agrobacterium EHA105 are commonly used strains, most molecular biology laboratories have preservation, and the rice variety is wild japonica rice medium flower 11 (a publicly used rice variety, which is commercially available).

The main reagents used In the examples were restriction enzymes, Taq enzyme, T4 ligase, Pyrobest Taq enzyme, KOD, RNase A, 5 XAll-In-One RT MasterMix reverse transcription kit and the like available from organisms such as TAKARA (Dalian), Promega, NEB, ABM and the like, dNTPs available from Genestar, plasmid miniprep kit and agarose gel recovery kit available from Shanghai Czeri bioengineering, TR L zol RNA extraction kit available from Invitrogen, MS medium, agar powder, agarose, ampicillin (Amp), kanamycin (Kan), gentamicin sulfate (Gen), rifampicin (Rif) and the like antibiotics and the like available from Sigma.

Example 1 analysis of phenotype and genetic Effect of Rice esp mutants

A plant which is slightly short, has obviously shortened ear length, is upright in ear shape and has increased grain density is screened from the progeny of the planted wild japonica rice mid-flower 11, and the plant is named as a rice esp mutant.

In grain filling stage and mature stage, plants and panicle types of the wild type and the mutant heterozygote are photographed respectively, the phenotype is shown in figures 1A and 1B, the panicle of the esp heterozygote mutant is shorter and the panicle density is increased compared with the wild type (figures 1A and 1B). Interestingly, the mutation has a low frequency of spontaneous reversion mutation phenomena, so that different tillers of individual plants and even different ear branches of the same rice ear can present the phenomenon of coexistence of wild type and mutant type. The phenotype of the esp homozygous mutant and the wild type at the seedling stage is shown in FIG. 1C, and the plant of the esp homozygous mutant is short, short in leaves and weak in apical dominance. At the early stage of ear development, ear development of the esp homozygous mutant was severely affected, and the ear tip was stunted, necrotic, resulting in failure to form a viable ear (fig. 1D and 1E).

In order to analyze the genetic characteristics of the esp mutant, the characteristics of 210 plants of selfed progeny of the mutant were investigated and analyzed and divided into 3 phenotypes, wherein 58 plants are similar to the wild-type phenotype, 103 plants show the short-ear and dense-ear phenotypes (heterozygous mutant), 49 plants show the dwarf phenotype (homozygous mutant), and the segregation ratio of the three phenotypes is 1:1.78:0.84 (1: 2:1, chi-shaped phenotype is satisfied)²＝0.85，P>0.05). The above results indicate that the esp mutant phenotype belongs to a single gene-controlled incomplete dominant mutation.

Example 2 analysis of ear traits of esp mutant

The esp heterozygous mutant and the wild rice panicle before the completion of the filling to the yellow maturity are photographed, and the panicle characters such as panicle length, grain type, the number of branches per time and the length of branches per time are statistically analyzed, and the results are shown in fig. 2. The panicle length of the ESP heterozygous mutant is significantly shortened (fig. 2A and 2B), the length of the primary branch is also significantly shortened (fig. 2C), and the grain type and the number of the primary branches are not significantly changed (fig. 2A and 2D), which shows that the ESP mainly influences the development of the primary branch of the rice panicle, further influences the panicle type and the panicle length, and basically has no influence on the number of the primary branches and the grain type.

Example 3 map-based cloning of ESP Gene and analysis of candidate Gene expression

F constructed by using esp mutant and indica rice 74 (publicly used rice variety, commercially available)₂Population (esp is hybridized with Hua-nong-nonglutinous indica 74 to obtain F₁，F₁Selfing to obtain F₂Population), ESP was genetically mapped between SSR markers RM583 and RM9 on chromosome 1, with a genetic distance of 42.4 cM. An Indel (insertion-deletion) marker was developed between these two markers according to the japanese sunny and 9311 reference sequencesThe ESP gene was further located in the 51.2kb region of rice chromosome 1 (FIG. 3A). Within this interval there are 4 genes, Os01g0356900, Os01g0356951, Os01g0357100 and Os01g 0357200.

In order to identify the candidate gene ESP, the expression levels of the 4 genes in the ESP mutant and the wild type are respectively detected, and the specific steps are as follows:

(1) the Trizol method extracts RNA from the wild type and the esp mutant.

(2) Reverse transcription first strand (cDNA) synthesis was performed using 5X All-In-One RT MasterMix reverse transcription reagent from ABM, according to the instructions of the product.

(3) L una Universal qPCRMaster Mix reagent of NEB company is used for carrying out fluorescent quantitative PCR detection, and 25sRNA gene is selected as an internal reference gene to finish the operation according to the product instruction.

The primer sequences used were as follows:

Os01g0356900F(5′-3′):CATCGAGATCAAGGTGAGCA；

Os01g0356900R(5′-3′):GAAGTCGTTGAACCGGATGT；

01G0356951F(5′-3′):ATCGCTATCTCGCCGAGTC；

01G0356951R(5′-3′):CAGCAAGTCTCCACAGTCCA；

Os01g0357100F(5′-3′):GGAGGACCTCATCGACAAGA；

Os01g0357100R(5′-3′):GTTCGGGATCACGATGTTCT；

Os01g0357200F(5′-3′):GGCTTCGTCTACCAGCTCAA；

Os01g0357200R(5′-3′):CCCCTTGATCTTCAATTTGC；

25sRNAF(5′-3′):AAGGCCGAAGAGGAGAAAGGT；

25sRNAR(5′-3′):TTGGCGGGCCGTTAAGCAGAAAAGA。

the fluorescent quantitative PCR reaction program is as follows:

the results of expression analysis of 4 genes in the wild type and the esp mutant showed that there was no significant difference in the expression level of 3 genes, Os01g0356900, Os01g0357100 and Os01g0357200, in the esp mutant compared to the wild type (fig. 3B), the Os01g0356951 gene was hardly detected in the wild type, but its expression level was significantly up-regulated in the esp mutant, and the expression level of the gene was higher in the homozygote than in the heterozygote (fig. 3C). The results of the above gene expression analysis indicate that Os01g0356951 may be a candidate gene for ESP.

The expression level of Os01g0356951 of mutant plants with normal phenotype rice ears and mutant plants with mutant phenotype rice ears are named as mutant chimera plants, and the result shows that the expression level of the Os01g0356951 gene of tillers with erect dense ear phenotype of the chimera is higher than that of tillers with normal phenotype on the same plant (figure 3D). In addition, tracking detection shows that all plants with erect dense spike phenotype in the selfing progeny of the chimeric mutant have the characteristic of high expression of the Os01g0356951 gene, and the expression of the Os01g0356951 gene of the plants with normal phenotype (Epi-normal) is not different from that of the wild type (japonica rice middle flower 11) (FIG. 3E). The results show that the erect dense spike phenotype in the mutant is caused by the overexpression of the Os01g0356951 gene, and further support that the Os01g0356951 is a candidate gene of the ESP.

Example 4 methylation analysis of mutant and wild-type ESP genes

In order to search mutation sites, the invention designs primers according to rice genome sequence data, amplifies target segments of wild type and mutant, and sequencing analysis shows that no change of DNA sequence is found in a 51.2kb positioning interval, but the expression quantity of ESP candidate genes has extremely obvious difference, so that the target segments are presumed to have epigenetic variation. To this end, the present invention analyzes methylation modification variation by bisulfite sequencing, which is described in the literature (Zhang, XQ, Sun J, Cao X, Song X. epigenetic mutation of RAV6affects leaves and feedstock in rice. plant Physiol,2015,169(3): 2118. sup. 2128), and the procedure is briefly as follows:

(1) wild type (flower 11 in wild type) and esp mutant genomic DNA were treated with bisulfite, after which all unmethylated cytosines were converted to uracil, whereas methylated cytosines did not change.

(2) Designing a primer, and carrying out PCR amplification by using the genomic DNA before treatment as a template and using ESP-TTRgF and ESP-TTRgR; PCR amplification was performed using the treated genomic DNA as a template and ESP-TTRgF and ESP-TTRBSR. The primer sequences are as follows:

ESP-TTRgF(5′-3′)：GGTGATAATTTTGGAGGTTGTAG；

ESP-TTRgR(5′-3′)：TGCAGGATTGAATTCCTGATAAACT；

ESP-TTRBSR(5′-3′)：TACAAAATTAAATTCCTAATAAACT。

(3) and purifying the target fragment, performing TA cloning, and selecting positive clones for sequencing.

(4) Sequences determined by bisulfite treatment were aligned to untreated sequences, the methylation sites and numbers were counted, and the wild type and mutants were analyzed for methylation differences.

As a result, DNA demethylation occurred in the downstream of the 3' end of the ESP gene (FIGS. 4A and 4B), and the specific methylation mutation site occurred in the 289-601 bp region of the downstream of the ESP gene (FIG. 4B).

Many studies have shown that DNA demethylation can often lead to overexpression of a gene (L i, et al, single-base resolution maps of mutated and wild-type genes and regulations of DNA methyl in plant gene expression 2012,13: 300).

To further elucidate the relationship between ESP gene expression and methylation modification, the present invention treated the germinated seeds of flower 11 in the wild type with the methylation inhibitor 5-azadeoxycytidine (5-aza-2' deoxycytidine, 5-aza-dC), the methylation inhibitor treatment method was described in the literature (Chang S, Pikaard CS. Transcript profiling in Arabidopsis thaliana variants. J Biol Chemi,2005,280:796-804), and then 7-day-old seedlings were taken for expression analysis of the genes (primers for expression analysis of ESP were 01G0356951F and 01G 0356951R). The results show that the expression level of the ESP gene was significantly up-regulated in seedlings treated with the methylation inhibitor 5-aza-dC compared to seedlings not treated with 5-aza-dC (FIG. 4C), suggesting that demethylation of the downstream segment of the ESP gene causes overexpression of the ESP gene, which is a key factor in regulating ESP gene expression, leading to mutant phenotype.

Example 5 functional complementation assay

In order to verify the function of an ESP gene, the invention constructs an ESP overexpression vector driven by a maize ubi (ubiquitin) promoter by utilizing a pCubi1390 vector, names pCubi1390-ESP-OE, transforms a rice variety middle flower 11 through mediation of agrobacterium EHA105, and performs ESP function complementation verification, and the steps are as follows:

(1) the ESP gene sequence (SEQ ID NO: 1) was used as a template to design the following primers:

1390-ESP-KpnI-F：5′-GGGGTACCAGATCGAAAAAATCGCTATCTC-3′；

1390-ESP-BamHI-R：5′-CGGGATCCCTAGTTCTTTTAAAAATTTG-3′。

(2) PCR amplification of ESP gene is carried out by using primer pair, obtained target gene fragment is cut by KpnI and BamHI, recovered and connected to linearized pCUbi1390 vector recovered after cutting by the same enzyme to form pCUbi1390-ESP-OE vector, and sequencing is carried out, wherein DNA L alignment KitVer.2.0 kit is used for connecting ESP gene and expression vector pCUbi1390 after cutting by double enzyme.

(3) Agrobacterium EHA105 was transformed with the correctly sequenced pCUbi1390-ESP-OE vector.

(4) The agrobacterium EHA105 strain containing recombinant plasmid pCUbi1390-ESP-OE is transformed into the callus of wild type medium flower 11, and the callus with hygromycin resistance, differentiation, rooting, seedling hardening and transplanting are screened to the field to obtain transgenic plants after preculture, dip dyeing and coculture.

The specific method of rice transgenosis refers to the application of Chinese invention patent CN201510009631.4 and rice B3 transcription factor family gene RAV 2.

The result of the functional complementation test is shown in FIG. 5, wherein, the overexpression vector pCUbi1390-ESP-OE of the ESP gene is transformed into the wild type middle flower 11 to obtain the transgenic positive strain OE, the rice ear of which is shortened, the grain density is increased, the ear is upright, and the phenotype is similar to that of the mutant. The results of the above studies indicate that the ESP mutant phenotype is indeed caused by overexpression of the ESP gene. In addition, this example also shows that the method for obtaining the vertical dense-ear rice variety by up-regulating the ESP gene expression has strong operability.

In summary, the present invention concludes: the ESP gene transcription is activated to generate mutation phenotype due to DNA methylation modification variation of the adjacent segment at the downstream of the 3' end of the ESP gene; the ESP gene has an important negative regulation effect on the rice panicle type; the method has strong operability on rice panicle genetic improvement by regulating the expression level of the ESP gene.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Sequence listing

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Claims (10)

1. Gene for regulating development of rice spikeESPThe application in adjusting the development and the spike shape of the rice spike is characterized in that:

the gene for regulating and controlling the development of rice earsESPIs SEQ ID NO: 1.

2. The gene for regulating development of rice panicle according to claim 1ESPIn regulating development and panicle type of riceThe application of (2), which is characterized in that:

gene for regulating development of rice spikeESPThe application in the rice ear type improvement breeding.

3. The gene for regulating development of rice panicle according to claim 1ESPThe application in adjusting the development and the spike shape of the rice spike is characterized in that:

gene for regulating development of rice spikeESPThe application in cultivating upright dense-ear rice variety.

4. The gene for regulating development of rice panicle according to claim 2ESPThe application in adjusting the development and the spike shape of the rice spike is characterized in that: the rice panicle type improved breeding mode comprises transgenic breeding and crossbreeding.

5. The gene for regulating development of rice panicle according to claim 4ESPThe application in adjusting the development and the spike shape of the rice spike is characterized in that:

the transgenic breeding is to containESPThe recombinant expression vector of the gene is introduced into plant cells by changingESPObtaining the spike variation transgenic rice by the expression level of the gene;

the hybrid breeding is to improve the panicle type of rice by using the panicle type mutated transgenic rice as a donor parent and a hybrid breeding method, namely, to cross the obtained panicle type mutated transgenic rice with a rice variety to be improved and then breed the panicle type improved rice variety;

said composition containsESPThe recombinant expression vector of the gene is constructed by the existing plant expression vector.

6. The gene for regulating development of rice panicle according to claim 5ESPThe application in adjusting the development and the spike shape of the rice spike is characterized in that: in the construction of said compound containingESPWhen recombinant expression vectors of genes are usedESPThe transcription initiation nucleotide of the gene is added with any one of enhancement type, composition type and tissue specificityOr an inducible promoter.

7. The gene for regulating development of rice panicle according to claim 5ESPThe application in adjusting the development and the spike shape of the rice spike is characterized in that: the plant expression vector is used after being modified, and antibiotic resistance genes are added.

8. Gene for regulating development of rice spikeESPThe recombinant expression vector, the transgenic cell line or the recombinant strain are applied to the regulation of the development and the panicle type of the rice panicle, and are characterized in that:

the gene for regulating and controlling the development of rice earsESPIs SEQ ID NO: 1.

9. Use according to claim 8, characterized in that:

gene for regulating development of rice spikeESPThe recombinant expression vector, the transgenic cell line or the recombinant strain in the rice panicle type improvement breeding.

10. Use according to claim 8, characterized in that:

gene for regulating development of rice spikeESPThe recombinant expression vector, the transgenic cell line or the recombinant strain of (2) are applied to the cultivation of the vertical dense-ear rice variety.

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