CN110240641B - Rice DPS1 gene and application of coded protein thereof - Google Patents

Abstract

The invention provides a rice DPS1 gene, and a coded protein and application thereof. The rice DPS1 gene sequence is shown as SEQ ID No.2, and the coding protein sequence is shown as SEQ ID No. 1. The invention discovers that the tip spikelet is shriveled and necrosed before the mutant of the rice DPS1 gene blooms, and the middle spikelet close to the tip degenerated spikelet is sterile, so that the seed setting rate and the spike grain number are remarkably reduced. The wild DPS1 gene is transformed into the rice DPS1 mutant, the mutant ear top degeneration phenotype can be recovered, and the DPS1 gene is shown to directly control the ear top degeneration by regulating cell programmed death, can be applied to the oriented design of the ear type of rice, can be used as an important gene resource, is beneficial to developing a molecular marker in molecular breeding practice to carry out early identification and elimination on the ear top degeneration, and has important guiding significance for improving the rice yield.

Description

Rice DPS1 gene and application of coded protein thereof

Technical Field

The invention belongs to the field of genetic engineering, and particularly relates to a rice DPS1 gene, a coded protein and application thereof.

Background

The rice is one of the most important grain crops in China and even the world, more than 60% of people in China use the rice as staple food, and the demand on the grain is increased along with the acceleration of the urbanization process, the reduction of the cultivated land area, the application of biological energy and the like. Therefore, increasing rice yield is of great significance to solve the increasingly serious food problems. The rice ear yield is the most basic unit for forming the rice yield, and the large ear grain is always one of the important targets of rice breeding in the world. However, the degradation of the top of the spike is a common biological phenomenon in rice breeding and field production, so that the reduction of the volume of the spike and the number of grains per spike are obviously reduced, and the yield per spike of rice is seriously influenced. Due to the complex genetic basis of the ear tip degeneration, the ear tip degeneration is easily influenced by environmental conditions such as temperature and humidity during the ear differentiation, the phenotype identification is difficult or unstable, and the molecular mechanism of the ear tip degeneration is still poorly understood at present. Therefore, the novel gene for controlling the rice ear top degeneration is separated, the action mechanism of the novel gene is researched, the genetic control mechanism of the ear top degeneration is favorably clarified, the yield loss caused by the ear top degeneration in breeding and production practices is overcome, and the novel gene has important theoretical and practical significance for high-yield and stable-yield breeding of rice.

The main factors of rice yield include the ear traits such as effective ear number per unit area, grain number per ear, thousand grain weight and seed setting rate. The formation of the rice spike starts from a shoot tip meristem, and after the rice spike is converted into a spike stalk meristem, the rice spike is differentiated through a primary branch and a secondary branch to finally form a spikelet. Although there is currently no uniform standard for division of the development stage of spikelets, the process is essentially the same. When the external environmental conditions and internal factors are suitable for flower formation induction, a plurality of small bulges are formed on a apical meristem (SAM), and the stem apical meristem SAM is changed into an Inflorescence Meristem (IM) rice to enter a reproductive growth stage. Some small bulges turn into Primary Branch Meristems (PBM) and the main cob of the inflorescence; after a period of time, the apical meristem of the inflorescence stops developing to form a degenerate point, followed by the formation of a primary shoot meristem to form a secondary shoot meristem (SBM) and a Spikelet Meristem (SM), which develop into a shoot and a spikelet, respectively. Finally, glume-protecting, palea and lemma, and flower organ primordium such as stamen and pistil are successively formed on the spikelet meristem. After the spike structure is formed, the rice enters a germ cell forming stage, the development of female germ cells (embryo sacs) and the development of male germ cells (pollen) are carried out simultaneously, and a complete spikelet is formed after the process is completed. With the research and development of molecular biology, more and more genes related to development of rice spikelet are currently identified and cloned, many of these genes are involved in the regulation of various meristem transitions and initiation, such as axillary meristem initiation regulation genes LAX1, LAX2, MONOCULM1(MOC1), etc., meristem differentiation time regulation genes APO1(ANICLE ORGANIZATION 1), TAW1 (TAWAWAWAWAWAWAWA 1), FZP (frizzy panicle), etc., and additionally hormones such as auxin and cytokinin have important regulation effects on the activity of meristem during spikelet development.

The ear branch number of rice directly determines the ear grain number, and the ear grain number is a key factor directly related to the rice yield. However, when the rice spike is in the development stage of florets, the formed florets are easy to degenerate, which is mainly characterized in that glume flowers and branches at the back of the spike degenerate and whiten, and the spike aborts and falls off after being mature. The rice ear degeneration mainly comprises two types, namely ear degeneration at the base of the ear and ear degeneration at the top of the ear. The environmental factors influencing the degradation of the rice spikelets mainly comprise the external temperature, humidity, illumination, the change of nutrient conditions in soil and the like. The stratum corneum development defect and active oxygen accumulation are two important physiological factors causing the sterile degeneration of spikelets, and the stratum corneum not only has the protection effect on external stimulation, but also serves as a selective permeable membrane for exchanging water and nutrient substances to avoid excessive water loss. In addition, it affects cell-to-cell communication by modulating the pathway of signaling molecules. Cuticle-deficient mutants often exhibit partial or complete sterility, thus resulting in a decrease in seed set rate. Abnormal accumulation of Reactive Oxygen Species (ROS) in plants can also cause damage to cells and their organelles, leading to programmed cell death. The tip floret degenerated mutant paab1-1 accumulates too high hydrogen peroxide and MDA at the degenerated part. However, little is currently known about the genetic control and molecular mechanisms of spikelet degeneration.

Regarding the genetic mechanism research of the rice panicle base, Yamagishi and the like find 3 main effect QTLs related to the abortion of florets at the rice panicle base before flowering on the 1 st, 10 th and 11 th chromosomes by using backcross inbred line populations of the temperate japonica rice Akihikari and the African upland rice IRAT109 as materials. Li et al (2009) clone a spike basal degeneration-related gene SP1, wherein the gene encodes a transport protein containing a PTR (protein transport receptor) domain, the expression level in phloem of young spikes is high, the mutant primary branch primordium of the gene can be normally initiated and differentiated, but the elongation of the primary branch of the base is seriously hindered, so that the basal branch and florets are degenerated and died, and finally the mutant shows that the weight of the spike and thousand grains is reduced, but the specific molecular mechanism of SP1 for regulating the basal glume degeneration is not clear. Phylogenetic analysis shows that the homology of SP1 and nitrate transporter AtNRT1.2 in Arabidopsis is high, so SP1 is presumed to supply nitrogen to young ears through transporting nitrate, thereby maintaining the normal growth and development of young ears.

Compared with the degradation of the spikelets at the base of rice, the degradation of the spikelets at the top of rice is more common in agricultural production. For example, the ear degeneration rate of the early high-yield variety agricultural cultivation 58 reaches 15% -20%, which seriously affects the rice yield, and the excellent indica rice variety 9311 which has been sequenced and widely applied in production also has a serious ear top degeneration phenomenon. In recent years, Quantitative Trait Loci (QTLs) associated with head degeneration have been reported in succession. Xuhuashan et al (2007) detected 5 QTLs on chromosomes 1, 2, 4, 7 and 9 using the japanese sunny and 9311 replacement line population, where the ds-9 locus on chromosome 9 had a relatively large effect on spikelet top spikelet degeneration. Tan et al detected a qPDS3/qMDS3 locus on chromosome 3, which contributed 11% to the retrogression of the apical spikelet, and the locus and the QTL locus on chromosome 9 had an interactive effect in regulating the retrogression of the apical spikelet. And et al found 6 QTLs controlling tassel top degeneration on chromosomes 1, 2, 3, 5, 6 and 7 using a recombinant inbred line population. QTL sites associated with the degeneration of the apical spikelet were detected on chromosome 4 in Zijuan san et al. Homosowei and the like found 5 QTLs for controlling the top degeneration of the panicle by using a panicle top degeneration mutant L-05261, distributed on chromosomes 3, 4, 5 and 8, and found that a large amount of hydrogen peroxide substances are accumulated in the degenerated spikelets by using 3,3' -Diaminobenzidine (DAB) detection, presumably being a main cause of the top spikelet degeneration. Cheng et al found a spike top degeneration QTL qPAA8 affected by lighting conditions and localized it within the range of 68kb, and also found that spike top degeneration was associated with the accumulation of hydrogen peroxide. Active oxygen is used as a signal molecule, the amount of the active oxygen in a plant body is strictly regulated, and the active oxygen can cause male sterility of rice under the condition of excessive accumulation.

Because the degradation of the top of the ear is greatly influenced by environmental factors, the difficulty in finely positioning QTL sites and separating candidate genes is high. In recent years, researchers at home and abroad use mutants to identify a series of candidate genes for regulating and controlling top spikelet degeneration. Qi et al (2013) report that OsPUP7 gene mutation results in rice plant height increase, heading period extension and complete degeneration and sterility of top spikelet. Chemically mutagenizing a Korean japonica rice variety Hwachengbyeo to obtain a spike tip degeneration mutant paa-h, wherein the mutant plant is dwarf, small in kernel, small and upright in spike shape, and degenerates the top spikelet, so that the spike grain number and thousand kernel weight are both remarkably reduced. Through a map-based cloning method, a candidate gene is positioned in a 71kb interval on a 4 th chromosome, and sequencing finds that the coding region of the LOC _ Os04G56160 gene in the interval has a G-to-A base mutation, so that glycine is changed into glutamic acid. However, the molecular mechanism of the gene for regulating the degeneration of the apical spikelet is still to be further researched. The rice ear degeneration mutant tut1(tutou1) shows that the root is shortened, the cells at the top end of the leaf are dead, the plant height is reduced, the tillering is reduced, and the like, and the most remarkable phenotype is the defect of the tip spikelet degeneration of the inflorescence. The TUT1 gene is located on chromosome 1 by using a map-based cloning method, and has expression in spikelet, seed bud, root, coleoptile, stem node and leaf, and the expression level is highest in floral organs. The gene codes cAMP receptor protein inhibitor and can promote actin nucleation and in vitro polymerization. In addition, the tut1 mutant ear tip degeneration phenotype is closely related to the disintegration and defect of the cuticle, suggesting that the formation of the cuticle affects the development of the rice ear. Recently, the ear tip degenerated gene OsALMT7(aluminum-activated plate transporter) and SPL6 (squarosan promoter-binding protein-like) were cloned successively. The OsALMT7 gene is obtained by cloning a panicle top floret degenerated mutant paab1-1 by a Wanjian academy team (2018), and the panicle top degenerated mutant is a programmed cell death and DNA fragmentation process in the late growth stage of young panicles. The OsALM7 gene is located on chromosome 2 and encodes a plasma membrane protein with a function of transporting malic acid. The results show that the OsALMT 7-mediated malic acid transportation plays an important role in maintaining the normal growth and development of the rice ears. The SPL6 gene presents a top high-level expression pattern in the process of ear development, encodes a rice SBP-box family transcription factor, and inhibits the transcription of an endoplasmic reticulum stress induction factor IRE1(ER-stress sensor), thereby controlling the output intensity of a stress signal. The SPL6 gene deletion mutant apical spikelet presents obvious cell programmed death characteristics under normal growth conditions, and the transcription level and the protein level of an endoplasmic reticulum stress induction factor (ER-stress sensor) IRE1 are remarkably increased, so that the output of an endoplasmic reticulum stress signal is out of control and downstream gene overstimulation expression is caused, and the apical spikelet cell senescence degeneration and the spike baldness character are caused.

So far, although a plurality of top spikelet degeneration-associated genes have been cloned in rice, the molecular mechanism of top spikelet degeneration is not clearly analyzed, and a more complete and fine rice top spikelet degeneration molecular regulation network is to be constructed. In addition, a gene that can be effectively applied to reduce the rate of spikelet degeneration has not been discovered. Therefore, the separation of the mutant with stable spike top degeneration phenotype, the cloning of related genes and the clarification of the functions of the mutant have important guiding significance for further analyzing the rice spike top degeneration regulation mechanism and developing molecular markers in molecular breeding practice to early identify and eliminate the unfavorable character.

Disclosure of Invention

The invention aims to provide a gene capable of reducing the rice top spikelet degradation rate and application thereof.

The rice DPS1 protein provided by the invention has the following characteristics:

1) an amino acid sequence shown as SEQ ID No. 3; or

2) Protein which is derived from the protein 1) and has equivalent activity and is obtained by substituting, deleting and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID No. 3.

The present invention provides a gene encoding rice DPS1 protein, which has:

1) a nucleotide sequence shown as SEQ ID No. 2; or

2) The nucleotide sequence shown in SEQ ID No.2 is substituted, deleted and/or added with one or more nucleotides; or

3) Nucleotide sequences which hybridize under stringent conditions with the DNA sequences defined in 1).

The biological material containing the gene belongs to the protection scope of the invention, and the biological material is a vector, a transgenic cell line, an engineering bacterium, a host cell or an expression cassette.

The invention obtains a rice tip degeneration mutant dps1(degenerated panicle and spike 1, dps1) by screening through a method of inducing japonica rice Sanchi rice 16 by EMS.

The invention discovers that the splice site of the 11 th exon and the 11 th intron of the DPS1 gene in the mutant DPS1 mutant has a single base mutation from G to A (the mutation is positioned at the 7427 th nucleotide of SEQ ID No. 1) by comparing the mutant with the DPS1 genome sequence of the Saint rice 16. Based on this mutation, one skilled in the art can develop molecular markers for breeding assistance based on this site.

In the early stage of growth and development, the vegetative growth and reproductive growth of the dps1 mutant are normal, have no obvious difference compared with a wild type, and continue until young spikes develop to about 10 cm; when the young spike reaches about 12cm, the young spike at the top of the mutant spike is pale and stagnates in development, and obvious degradation phenomenon occurs; with the further development of the rice ears, the degraded small ears at the top end continuously increase, and when the ears reach the final length before the flowering period, the degradation phenomenon becomes more obvious and is expressed as shrivelled necrosis, the degraded small ears account for about 10% of the whole ears, the middle small ears close to the degraded small ears at the top are also affected and are expressed as sterile, and the maturing rate and the ear number are extremely reduced obviously. The dps1 mutant has reduced activity of antioxidant system enzymes SOD and CAT in the top spikelet, and hydrogen peroxide accumulation, and programmed cell death characterized by nuclear DNA damage occurs.

The invention provides application of the rice DPS1 protein or the coding gene thereof or the biological material containing the gene in preparing transgenic plants.

The invention provides application of the rice DPS1 protein or the coding gene thereof or the biological material containing the gene in rice panicle type improvement or genetic breeding.

The invention provides application of the rice DPS1 protein or the coding gene thereof or the biological material containing the gene in improving the rice setting percentage, the grain number per ear or the rice yield.

The invention provides application of the rice DPS1 protein or the coding gene thereof or the biological material containing the gene in restoring the apical spikelet fertility.

The invention provides application of the rice DPS1 protein or the coding gene thereof or a biological material containing the gene in improving the activities of enzymes SOD and CAT of a spikelet antioxidant system at the top of a spike of rice or improving programmed cell death of the spike of rice.

The invention provides application of the rice DPS1 protein or the coding gene thereof or the biological material containing the gene in reducing the rice spike top degeneration.

The invention further provides a molecular marker related to the rice top spikelet degeneration trait, the molecular marker is an SNP molecular marker of nucleotide 7427 of SEQ ID No.1, and the polymorphism of the SNP molecular marker is G/A, wherein single base mutation from G to A occurs at the splicing site of the 11 th exon and the 11 th intron of the DPS1 gene.

Based on the application of the DPS1 gene provided by the invention, a person skilled in the art can understand that the molecular marker of the gene can be used for breeding rice, screening high-yield rice or eliminating degraded rice at the top of ears.

The DPS1 gene has the function of regulating the rice top spikelet degeneration, can reduce or reduce the incidence rate of the rice top spikelet degeneration, further improves the rice yield, can be applied to the directional design of the rice spike type, can be used as an important gene resource, is beneficial to developing a molecular marker in molecular breeding practice to carry out early identification and elimination on the spike top degeneration, and has important guiding significance for improving the rice yield.

Drawings

FIG. 1 shows the phenotype of dps1 mutant and wild-type san Dioscorea rice 16.

FIG. 2 is a diagram showing the location and structure of the DPS1 gene.

FIG. 3 is the structural diagram of vector pCAMBIA 1305.1: DPS 1.

FIG. 4 shows the structure of vector pCAMBIA1305.1-APFHN DPS 1.

FIG. 5 shows that both pCAMBIA 1305.1:: DPS1 and pCAMBIA1305.1-APFHN:: DPS1 transformed rice DPS1 mutant can restore its phenotype.

FIG. 6 is an analysis chart of the expression pattern of the DPS1 gene in various rice tissues.

FIG. 7 shows that the rice dps1 mutant exhibited reduced activity of hydrogen peroxide accumulation in the apical spikelet and antioxidant system, and programmed cell death characterized by nuclear DNA damage.

FIG. 8 is a diagram showing the comet analysis of the cells of the spikelet at the tip of the dps1 mutant.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

The rice (Oryza sativa) variety Saint Rice 16 used in the following examples was a standard variety, and the rice panicle top degeneration mutant dps1 was obtained from the institute of crop science, Chinese academy of agricultural sciences.

Example 1 acquisition and phenotypic analysis of mutants

The japonica rice san-dao 16 was mutagenized by Ethyl Methanesulfonate (EMS), and a spike tip degeneration mutant dps1(degenerated panicle and spike 1, dps1) was obtained by screening (FIG. 1, A). In the early stage of growth and development, the vegetative growth and reproductive growth of the dps1 mutant were normal, with no significant difference compared to the wild type, and continued until young ears developed to around 10cm (fig. 1B, fig. 1C); when the young ear reaches about 12cm, the young ear at the top of the mutant ear is pale and stagnated in development, and obvious degeneration phenomenon appears (figure 1D, figure 1E); as the ear of rice further develops, the tip degenerated spikelet continues to increase, and this degeneration becomes more pronounced as the spike reaches final length before anthesis, manifested as shriveling and necrosis (fig. 1F).

Statistical results show that the number of degenerated glume flowers per ear tip of the dps1 mutant accounted for approximately 10% of the total ears, resulting in a significant reduction in the number of spikelets per ear of the dps1 mutant compared to the wild type (japonica rice san. oryzae 16) (WT: 226. + -.22; dps1: 171. + -.13) (G of FIG. 1, H of FIG. 1).

Further examining the development of the middle spikelet close to the tip degenerated spikelet, the results show that the middle part of the spike is affected and shows sterility, while the growth of the base spikelet is good, which finally results in the spike grain number (WT:214 + -20; dps1:66 + -10) and the setting rate of the dps1 mutant being significantly reduced (WT: 94.91% + -1.97%, dps1: 38.35% + -5.2%) (I in FIG. 1, J in FIG. 1). The above results indicate that the degradation of the tassel top of the dps1 mutant has a serious adverse effect on agronomic traits.

Example 2 acquisition of the Rice DPS1 Gene

The dps1 mutant was hybridized with phenotypically normal indica variety Huanghuazhan, F₁Normal in the case of F₂In the generation segregating population, the normal individual plant and the mutant individual plant meet the segregation ratio of 3:1, thereby indicating that the mutation character is controlled by a pair of recessive genes. With F₂The 30 mutants of (2) are used as materials, and the candidate genes are positioned between Indel markers M1 and M7 of chromosome 5 by using 170 Indel markers uniformly distributed on 12 chromosomes of rice. For fine localization of the gene, F₂The population was expanded to 472 mutant individuals and the candidate genes were located in the 28.85kb range between INDEL markers M3 and M5 (A of FIG. 2). The interval contains 6 open reading frames, wherein the gene sequence with the number of Os05g0395300 has gene function related to phenotype, and for this purpose, the full-length genome DNA of the gene is subjected to PCR amplification in 2 segments, each segment is about 3.6kb in size, the primers are shown in Table 1, and the sequencing results of wild type and mutant are analyzed by using DNAStar software. In the dps1 mutant, a single base mutation from G to A occurred at the splicing site of the 11 th exon and 11 introns of the gene (B in FIG. 2). RT-PCR results indicated that this mutation resulted in a missplicing of mRNA, resulting in two erroneous transcripts that differed from the normal wild-type (C in FIG. 2). The normal transcript of the gene in wild type was sequenced using the sequencing primers shown in table 1. The sequencing result shows that the result of the nucleotide sequence is shown as SEQ ID NO.2, and the result of the amino acid sequence is shown as SEQ ID NO. 3. The primer sequences referred to in example 2 are shown in Table 1Shown in the figure.

TABLE 1

Example 3 pcambial 305.1: : DPS1 vector transformed rice DPS1 mutant

In order to carry out a function complementation experiment, a DPS1 gene function complementation vector driven by a self promoter and an over-expression vector driven by a rice ACTIN1 promoter are respectively constructed. The DPS1 gene function complementary vector is driven by gene self promoter, 2011bp before ATG of translation initiation site is selected as gene promoter, and PCR amplification is carried out by taking genome DNA as template, wherein the used amplification primer is DPS1-gDNA-C shown in table 2. EcoRI site is introduced into the 5 'end, SpeI site is introduced into the 3' end, the length of PCR product is 9685bp (shown as a sequence in SEQ ID NO. 1), and finally the self promoter and the whole genome DNA (shown as a sequence in SEQ ID NO. 1) are cloned into pCAMBIAl305.1 vector to form a complementary vector driven by the self promoter (FIG. 3).

In addition, an overexpression vector was constructed using the plant binary expression vector pCAMBIAl305.1-APFHN, and PCR amplification was performed using cDNA as a template, using amplification primers such as DPS1-CDS-C shown in Table 2. Introducing NcoI site at 5 'end and SpeI site at 3' end, the PCR product length is 1587bp (shown as SEQ ID No.2 sequence), recombining into NcoI and SpeI sites of pCAMBIA1305.1-APFHN, driven by constitutive high expression rice Actin1 promoter. The constructed vector is shown in FIG. 4.

The constructed complementary vector and the overexpression vector are transferred into agrobacterium EHA105 by an electric shock method, rice dps1 mutant seeds are induced to callus to be used as receptor materials, and the rice is transformed by an agrobacterium-mediated transformation method. The functional complementation vector driven by its own promoter yielded 28 independent transformed lines, 22 of which were restored to the wild type phenotype (A of FIG. 5, C of FIG. 5, F of FIG. 5). Whereas the overexpression vector driven by the ACTIN1 promoter yielded 10 independent transformation lines, 8 of which were restored to the wild type phenotype (B in FIG. 5, C in FIG. 5, G in FIG. 5). In contrast to the wild type (D in FIG. 5) and dps1 mutant (E in FIG. 5), it was found that the transgenic lines no longer showed a retrogression of the top of the panicle, with both the seed set and floret architecture restored to wild type levels. These results indicate that it is indeed the mutation in the DPS1 gene that caused the tip-degenerated phenotype of the DPS1 mutant ears. The primer sequences referred to in example 3 are shown in table 2.

TABLE 2

Example 4 expression Pattern of Rice DPS1 Gene

The expression of DPS1 in tissues such as leaves, leaf sheaths, stems, roots, scions and anthers at different developmental stages is analyzed by using the Real-time PCR method, and as a result, as shown in FIG. 6, although DPS1 is expressed in all tissues, the expression level in each tissue is different, wherein the expression level is highest in the leaves, anthers and ears at the later stage of development, and the ear before flowering (P16) is significantly higher than the ears at other stages (P3, P8, P10 and P12), the expression pattern is consistent with the functional effect of the gene, and the top spikelet degeneration phenomenon of the ear before flowering of the DPS1 mutant is particularly obvious. The primer sequences referred to in example 4 are shown in table 3.

TABLE 3

Example 5 dps1 mutant Top spikelet Hydrogen peroxide accumulation and reduction of enzyme Activity of antioxidant System

Reactive Oxygen Species (ROS) is an important factor in causing programmed cell death, and studies have shown that Reactive oxygen species can induce cell death in plant and animal tissues. It is therefore speculated that an increase in reactive oxygen species levels in the tassel of the dps1 mutant leads to an increase in cell death. Hydrogen peroxide is a relatively stable active oxygen molecule, and DAB (3,3' -diaminobenzidine) staining was performed on the wild type and the mutant in this example to detect accumulation of hydrogen peroxide. The results show that dps1 mutant top spikelet glumes appear to be darker dark brown precipitate under DAB staining, while wild type top spikelet glumes under the same treatment are not significantly darker (a of fig. 7). Further taking the top spikelet tissues of the wild type and the mutant at different development stages, and carrying out quantitative analysis on the hydrogen peroxide content of the top spikelet tissues, wherein the results show that the hydrogen peroxide content is not different from that of the wild type in 6cm spikelets of dps1 mutant; this difference was even more pronounced when the young panicles reached 8cm, where the hydrogen peroxide content in the dps1 mutant began to be significantly higher than in the wild type, especially when the young panicles reached the 12cm, 15cm stage (fig. 7B).

This example also determined the level of Malondialdehyde (MDA), an important indicator of membrane lipid peroxidation by reactive oxygen species, and found that the MDA level of the mutant top spikelets was significantly higher than that of the wild type (fig. 7C). The experiments show that the spikelets at the tops of the mutants have active oxygen accumulation, and the reduction of the active oxygen scavenging capacity is also suggested, so that the activity of the enzyme of an antioxidant system is further measured. The results show a significant reduction in the activity of superoxide dismutase (SOD) and Catalase (CAT) in the mutant spikelets (D of fig. 7, E of fig. 7). It is speculated that defects in the reactive oxygen scavenging system in the dps1 mutant lead to reactive oxygen accumulation, which in turn leads to programmed death of the apical spikelet cells.

In order to further confirm that the dps1 mutant has DNA damage based on programmed cell death, the dps1 mutant apical spikelet cells were subjected to comet analysis, which can effectively detect and quantify the degree of single-and double-strand nick damage in the cells. After lysis of the cells of the desired tissue, staining with a fluorescent dye that binds to DNA, the cells with a high degree of DNA damage migrate to form a "comet" like image as chromosomal DNA migrates out of the nucleus, while the undamaged DNA portion remains spherical. Approximately 45% of the cells at the tip of the dps1 mutant spikelets showed high levels of DNA damage (50% tailing), 30% of 29% of the cells with 30% tailing, 10% of 14% with only 12% having the lowest level of damage (1% tailing). Whereas only 3% of cells in the wild-type top spikelet had a high degree of DNA damage (50% tailing), while most of the cells (43%) had the lowest degree of damage (1% tailing). It can be seen that programmed cell death, characterized by degradation of nuclear DNA, occurred in the dps1 mutant, which in turn resulted in the degeneration of the top spikelet (FIG. 8).

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

<110> institute of crop science of Chinese academy of agricultural sciences

<120> rice DPS1 gene and application of encoded protein thereof

<130> KHP191112936.3

<160> 31

<170> SIPOSequenceListing 1.0

<210> 1

<211> 9685

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

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cggacggcat ccgacggcca ccgtttccgt tccgcacgcc ccgtgctgct tttgctcttg 1860

gcgatttgcg gaagttgttg ttgtcccctt gctcctcgct ccgcttccat ccatattcca 1920

ccgcatcgcg cgcggggtta gggaggggcc tggtggtggt cggggctctg acgcggcggg 1980

gcggcggagg tggtggtggg gagggcgaga gatggctgcg gcggtgggga gggcgcgggg 2040

ggcggcgagc ctggcggcgg cggtgagggt ggcggcggcg acggcgaggc cggcgtcgag 2100

cgtcgccgcg gcggccgggc tggggttcgc ggggctggtc gtgcaggggg aggacacgcc 2160

gttcgggtcg gtgtggtggt gggcgtacgc ggggatatcc tgcttcctgg tgctcttcgc 2220

cgggatcatg tcggggctca ccctcggcct catgtcgctc ggcctcgtcg agctggagat 2280

cctccagcgc agcggcaccg atgccgagaa ggcccaggct ggtatgatcc ggaaccctag 2340

ccgcgttctt ctctcctccc ccgtgtcgtg atcgatctgg ttgcggtttg aggagccatt 2400

tctaggaggg tcttatgttg gaacagccca cttgtgctcg atctgatcgc aattttgcgg 2460

gcgattctta tggggtgttg agttgggata gcaggtgcta tgttacgtca cattggggat 2520

tcacgatatc tgtaacacgc catgattagt ttctgctatt ttggaatgag gatctattgc 2580

tgttattttg ctaaggattt gctgactctg atcgtgccgg tcatactcta gatcctctgt 2640

ttgttgtacc agttgtattt attttctatt ttaagttaac agaagcgaat atacatcaat 2700

aatgctgata atcagaaaaa gtagaacaca caaaagggta gtaaggttcc tctgccaaaa 2760

aggtagtaag gtgataagaa aaggagacgt tggtgggagt tggctttatt caagattcac 2820

ctctttctgg gggagatata aaatgatcct ttggtcagtg agaattggaa ctatgcaacg 2880

gataggcata gagatgtacg cgctagaagt tcctgggcct gccatgttgc ttgtgggtag 2940

cgtgcagctg agccttttca cacataagat atgatccata gttttttggt gcttttgttt 3000

gtactactaa ttttctgtga aatgatctag ttctgtctga aatgcaatat gggcatttta 3060

gatgttcacc atttgatcta atgtatatgc taaaattgat aacttgatat ggtagtagtg 3120

atggtgcttg tgtagtatta ttacaggcac tttcagcacc tcaataactc gttttcactt 3180

tttagcagat gctactgttg ctactgtatt ttcatggata gttctgttgt tgatcgtttg 3240

gctacttgta tgtcatcttt ttgtagctgc catcctccca gttgttcaaa agcagcacca 3300

gcttcttgtc accctgctct tgtgtaatgc ttgtgccatg gaggtatgta actctggttg 3360

catttctttt ttatcattta ccacactatt ttgtactgcg aacatgcaaa tataaagggt 3420

attagcagat gggcatattt gatagtatgt tattgttttg acacatttta actcatagct 3480

tgtttggaga gaagaatgct atatgctgaa gttgcctcta ttggttcagt tttgtctgta 3540

ctgcggcatg atttcctttg ttagcatctg tggaaatcca caaacgattt tacttgaatt 3600

gtaatatcta atttggaaga gtctagtgtc tttaattctt tgttaggtcc ctgaatgtaa 3660

ggtcatctac acttgatgat taatgctact ggcctactgc tacaagtcta tttgtcattg 3720

taatgtgcat taatatatct gagcatgtac cataaaatcc aacaaagtgg ctatattgaa 3780

gtttatatat caatattata taccttattt aaatatctag ttccctttaa tatttgaagt 3840

tctattaggt tttgcttgtt aactcgttat gtccataaaa aaaaatatgt tcaatctgct 3900

cacctactca ttaaatttgc taataatata acgcatttgg aaaattggaa ttgatttcac 3960

acattagaag ttctggtata tgattaggct cctataaagt ataaagtacg aatgtacggc 4020

ataagtggac tgctaagtgc tataacattt tctaggatcc tgagttctta acatgttctc 4080

tagatatatt attttctgct atgcacaaag gatgcccgag acaaatattt gtaaacttct 4140

ctttgcacat agtatggtat attagcttat tttacatgga tatgttttac cttcccccaa 4200

ccatggactt cctttcagaa ttttatcagt agatacacca tgcattctat tcatatacct 4260

tttacattgt tttgtttagc cagtgactct tccaatttgt ttctttggat cgtttgcagg 4320

ctcttcctat attccttgac aggatttttc atcctgttgt tgctgttatc ttatcagtaa 4380

cgtttgttct tgcttttgga gaggtattat atattatgtc caaactctat agtttctcta 4440

tggtcttgca gactgtctga ttgattattt aatttttaac gccaggttat accacaagca 4500

atctgtacta ggtatggatt ggctgtgggt gctaactttg tatggcttgt acgcatcctc 4560

atgatcatct gctatcctat ttcttatccc atagggaagg taagtccgca aaattaaatt 4620

gacttagttc aagtgtccct actataatga agtcttttgg agcttctcta atggcaggga 4680

cttttaattg tcaatttgtc atgaattaga tattgtaaca gttgtgcttc cgagtcatct 4740

cttcattcag ttttggtacc atagttgatg aaccctatta caaagtgaaa acaatatgcc 4800

ttcacaaaat gacctgtgta tatgtctttc gatttgagaa tttgttatca gcttatacta 4860

tattaggtta atcaatgccc ttagcttgta gtgctatgga agcttctatt actaaaacac 4920

catgcagcat gcaagctcat gtgattacaa atttgtgttt catgcattat gctataatgc 4980

agtgcatatg tttaaatgca cgcttaactt ggtgtacaaa cagtgtattt tctttcagat 5040

tttaatatgt tgtcttaggt ttaatgaaaa cagacttgat ttttgtacta tagaataaca 5100

tgtatatttt attttgtttt gaaatatttt ttgtttaagt cttccaaatt tacttgatat 5160

gatttaagga ctagttacta aaagtgattg aaatatatat tgtccatctg tcgcagctcc 5220

tagattgtgc tcttgggcat aatgaatctg cactttttag gcgagctcaa cttaaagctc 5280

ttgtctctat ccatagcaaa gaggtaacat gaccttacta tgtaacttat taataagtta 5340

acatcttctt tgaaggttat atcacagttc aatgcttttc tgccattgca ggctggcaag 5400

ggtggagagc ttacccatga tgagactaca atcataagtg gagccttgga cttgactgag 5460

aaggtgcgcg tcattttaaa atttgacttc taatttataa tgaataatag aatctggaga 5520

atcgttggca atcaaaattt aagtgaactt gttacaaagt tagatgttgt ttgacttgtt 5580

ttctagatat gacttgtttt ctatcttttg gttttagggg attgaaacat tagactccct 5640

attgtctttt atgtattaat ttgtattaat atttaactat gagatagctt ctagagtgat 5700

tttgatgtat atgatactga tatatttaat ggcttcatgc tttatggttg tcttcctatt 5760

tttgtttagt atttaacagc tttcagcttc acatcagaaa tgacttaatc catgctgcaa 5820

atattatatg acaaatgcct acatttgttc ttgtgtgttg aatctctttt tgactcttta 5880

ttgagcaaaa gaagtctgtt tcttacttta caaacatagt gctcacagac tagcactttg 5940

atatgaacaa tcattttttg ttgtcttcag cagaaatttc attgttattt gtgtgcagac 6000

tgctgaggag gctatgacac ctattgagtc aactttctca ttagatgtgg actccaagtt 6060

ggattggtaa ctataaaaaa aaatacaatc acactttaga caccgtagct cttggtcatg 6120

ttccatattt attatcgatc ctattcactg caaaagttct tgttgattct atgtttgtct 6180

tatgtctttc tttcttatta gttgaactca ttcgtcaaac gtaaacaatc ttctgtctta 6240

gttatgtatt tgtggaacta agttcagaca atgtaatctt ttgtgtttta cctacatagt 6300

agttattaaa cattagcaaa tagtacgcaa atttcccccc cttttttttt tgataaggct 6360

caatgctctg ccaatcttct tatgtacttt tatttacagg gaagcaatcg gcaaaatcct 6420

tgctcggggt catagccgtg tgcctgtata ctctggaaac cctaggaaca ttattggtct 6480

cctgctggtt ggtgtcgaat tgtcaatgct gtcatggagt atttaataac ccagtgactt 6540

gtttgtatga ttcaccatgt cttattttga atttttacag gtgaaaagtc ttctgacagt 6600

tcgtgctgaa acagagacac cagttagtgc tgtttccatt agaaggattc caaggtcgtg 6660

gcacaaatct attaaattat acattagaat gtcttctcaa catgggataa aatgttcata 6720

gttgaagtat cttttttgct gtattaatac ttgcaatctc cgtaatttct gacgaataat 6780

cagtcaatga ctttatttac cttctccagg gttcctgcag atatgccttt atatgacata 6840

ctgaatgagt ttcagaaagg aagtagtcat atggctgctg ttgtgaaggc taagcccaaa 6900

attgtaccac tccctgacaa aactgaacca aacagggaag taagtggggc accacagctg 6960

actgctccct tactatccaa taatgaagaa agggtggaaa gtttggttgt tgatattgaa 7020

aaaccacaga gcaggcaggt taatggaaac aaaccctgtt ccatgcagca gaatgagatg 7080

ccatatgcaa tgtctcggtc atcagaggat atagacgatg gtgaggtcat tggtatcatc 7140

acacttgaag atgtatttga agaactattg caggtaagct taagcatgct aaattttctg 7200

tctttaatag aactaaatcc tatcacaatt gcttatgctg tttttctttt tgttctttat 7260

caaaaagatc ctagccttgg cattaaagat gcattttggg atatagttgg aatcatttat 7320

acagttcaca attatgattg gatcttctgt atctcgtatg tttacgtttt attttacagg 7380

aggagatagt ggatgaaact gatgagtatg ttgatgttca taagaggtat gtctcatcgt 7440

tgcactttct cccctcattt tgcagtgttt gagagtatac tatgtgcgag taaattttct 7500

ctattacagg atccgagtgg ccgccgccgc cgctgcatct tcggttgcca gggctccatc 7560

aattcggaga ttaacgggtc aaaagggcac agtaagtgct attttttgct acagttttat 7620

agtcatattg gtcaccatga tggtataata acacaagtta cattgataca tgaataatat 7680

tgttgcataa atttttcact aggcactgag aatattggag tatgagatat atctttaatg 7740

ttgatcttta ggaatgatca tataagcttt ctggacaggg agggtaggta ctgctaaaaa 7800

tcttattacg tggattgagc tgatgtacct gacttcaaaa ctgtacaatc acctcaatca 7860

tactggggat tgaaatctgt tcatgttcat gtcaaaaaaa aaaaaatctg ttcacgtcaa 7920

gatttacctc gtttctgtta gttcaggctc tggcatggtc agtggcggat ccaggaatac 7980

ttttgagcct gggcaagtct aacagcgaaa aacaaatata cataaatcaa ttcaaatatg 8040

tatacctttt atcatatacg tatgtaaaat tgacggtcat tgagttctaa tttaggagag 8100

gaaatccagt gtatgagaat atatcaatga gatgatggtt atataggtta gactatggag 8160

tttgtactat cttattgaaa agaaaaaaga ctaataccag aaagttatat aattttgcct 8220

ttagatcaca gttgtgatgg tgtgaaattg aattaattat ctgaaagtaa taggcattta 8280

gttaagcatg gtaatcaaca agaagattca cgtgtaggag ggcattacct ataaattgta 8340

tgctgattag ctggtgacta aacactgtaa ctaagtgctg ataaactgct gcatagtggc 8400

ggagcccggg cacttgcccg cgctggccgg gccctagctc cgccactggg catggttgaa 8460

gttggattaa cttggtgttt gtttttccac ttggaaattg atcacaaaac tttgtaggga 8520

aatttgtttt aattgatttt tcagcaacaa taagctcaga tattttaatt tgaattaggt 8580

ggttccaatt ttagacatag ggctcttaga taagctgtgg gaattgcttg aagatctgca 8640

caagtgataa aatatttgct gagatcattt tcttttattt cttgctatat cattatgcat 8700

gtttgttttc tggttacact gattgtctga tttgtggaat atggtgtctt tggttgtata 8760

atataaaaga tatgaaactg tgcagttacg aacgatatgc ttacaatttt aaacatgtta 8820

acaagatgga cttgcaaaaa tttcttgaac catcgtccct aactgctact agcttagtac 8880

atatttatat tcatgtctga agatactgca tttggtccta actgtataaa cggttttaaa 8940

tcaaataacc ttgacccatt tttttttctt tatggtctat aaactgtttt gagtttccac 9000

atatttgtga tggaagtcct acttattgat gacgttactt tttgcctctt cagggtacgc 9060

agaataggca aggacagcca actggaattt tgaagaaacc tactgaaggt gattccaatc 9120

catcaaaaca tcaggtgaac cttgttgaac ctcttttgga aaataagagg taactttaga 9180

tgtatgcatg aatatagcga tggaacgcaa aatcaaattc agaaggcttg gctaacagta 9240

aatatgagcg aactcgtaaa ttcggatttg ttgtcatggt ctcatggacc ttagcgttga 9300

attttgtgtt tatggatgat gaaagtagat aacaattccg gaatcttttg atacatgaca 9360

gtagcaacga atagtactct tggaatctga aacgttgacc tggaggactg gaacggtagt 9420

ggatatggtt acattaggaa taagtacatt tcaaaggcta attgttcatg cgaaatatat 9480

attccagatg agactgcagc acaggtgaac acaatcggct gactgtttca attctgagaa 9540

acctggttca tccctggaaa ttttggaacc atttaccagt taccacgatt ccatcaattc 9600

cagttgcctg gcttcatgaa tttaccagtt atactcttca cctagttttt tttattccca 9660

tggaatagct agcgtatcta tctcg 9685

<210> 2

<211> 1587

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

atggctgcgg cggtggggag ggcgcggggg gcggcgagcc tggcggcggc ggtgagggtg 60

gcggcggcga cggcgaggcc ggcgtcgagc gtcgccgcgg cggccgggct ggggttcgcg 120

gggctggtcg tgcaggggga ggacacgccg ttcgggtcgg tgtggtggtg ggcgtacgcg 180

gggatatcct gcttcctggt gctcttcgcc gggatcatgt cggggctcac cctcggcctc 240

atgtcgctcg gcctcgtcga gctggagatc ctccagcgca gcggcaccga tgccgagaag 300

gcccaggctg ctgccatcct cccagttgtt caaaagcagc accagcttct tgtcaccctg 360

ctcttgtgta atgcttgtgc catggaggct cttcctatat tccttgacag gatttttcat 420

cctgttgttg ctgttatctt atcagtaacg tttgttcttg cttttggaga ggttatacca 480

caagcaatct gtactaggta tggattggct gtgggtgcta actttgtatg gcttgtacgc 540

atcctcatga tcatctgcta tcctatttct tatcccatag ggaagctcct agattgtgct 600

cttgggcata atgaatctgc actttttagg cgagctcaac ttaaagctct tgtctctatc 660

catagcaaag aggctggcaa gggtggagag cttacccatg atgagactac aatcataagt 720

ggagccttgg acttgactga gaagactgct gaggaggcta tgacacctat tgagtcaact 780

ttctcattag atgtggactc caagttggat tgggaagcaa tcggcaaaat ccttgctcgg 840

ggtcatagcc gtgtgcctgt atactctgga aaccctagga acattattgg tctcctgctg 900

gtgaaaagtc ttctgacagt tcgtgctgaa acagagacac cagttagtgc tgtttccatt 960

agaaggattc caagggttcc tgcagatatg cctttatatg acatactgaa tgagtttcag 1020

aaaggaagta gtcatatggc tgctgttgtg aaggctaagc ccaaaattgt accactccct 1080

gacaaaactg aaccaaacag ggaagtaagt ggggcaccac agctgactgc tcccttacta 1140

tccaataatg aagaaagggt ggaaagtttg gttgttgata ttgaaaaacc acagagcagg 1200

caggttaatg gaaacaaacc ctgttccatg cagcagaatg agatgccata tgcaatgtct 1260

cggtcatcag aggatataga cgatggtgag gtcattggta tcatcacact tgaagatgta 1320

tttgaagaac tattgcagga ggagatagtg gatgaaactg atgagtatgt tgatgttcat 1380

aagaggatcc gagtggccgc cgccgccgct gcatcttcgg ttgccagggc tccatcaatt 1440

cggagattaa cgggtcaaaa gggcacaggt acgcagaata ggcaaggaca gccaactgga 1500

attttgaaga aacctactga aggtgattcc aatccatcaa aacatcaggt gaaccttgtt 1560

gaacctcttt tggaaaataa gaggtaa 1587

<210> 3

<211> 528

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 3

Met Ala Ala Ala Val Gly Arg Ala Arg Gly Ala Ala Ser Leu Ala Ala

1 5 10 15

Ala Val Arg Val Ala Ala Ala Thr Ala Arg Pro Ala Ser Ser Val Ala

20 25 30

Ala Ala Ala Gly Leu Gly Phe Ala Gly Leu Val Val Gln Gly Glu Asp

35 40 45

Thr Pro Phe Gly Ser Val Trp Trp Trp Ala Tyr Ala Gly Ile Ser Cys

50 55 60

Phe Leu Val Leu Phe Ala Gly Ile Met Ser Gly Leu Thr Leu Gly Leu

65 70 75 80

Met Ser Leu Gly Leu Val Glu Leu Glu Ile Leu Gln Arg Ser Gly Thr

85 90 95

Asp Ala Glu Lys Ala Gln Ala Ala Ala Ile Leu Pro Val Val Gln Lys

100 105 110

Gln His Gln Leu Leu Val Thr Leu Leu Leu Cys Asn Ala Cys Ala Met

115 120 125

Glu Ala Leu Pro Ile Phe Leu Asp Arg Ile Phe His Pro Val Val Ala

130 135 140

Val Ile Leu Ser Val Thr Phe Val Leu Ala Phe Gly Glu Val Ile Pro

145 150 155 160

Gln Ala Ile Cys Thr Arg Tyr Gly Leu Ala Val Gly Ala Asn Phe Val

165 170 175

Trp Leu Val Arg Ile Leu Met Ile Ile Cys Tyr Pro Ile Ser Tyr Pro

180 185 190

Ile Gly Lys Leu Leu Asp Cys Ala Leu Gly His Asn Glu Ser Ala Leu

195 200 205

Phe Arg Arg Ala Gln Leu Lys Ala Leu Val Ser Ile His Ser Lys Glu

210 215 220

Ala Gly Lys Gly Gly Glu Leu Thr His Asp Glu Thr Thr Ile Ile Ser

225 230 235 240

Gly Ala Leu Asp Leu Thr Glu Lys Thr Ala Glu Glu Ala Met Thr Pro

245 250 255

Ile Glu Ser Thr Phe Ser Leu Asp Val Asp Ser Lys Leu Asp Trp Glu

260 265 270

Ala Ile Gly Lys Ile Leu Ala Arg Gly His Ser Arg Val Pro Val Tyr

275 280 285

Ser Gly Asn Pro Arg Asn Ile Ile Gly Leu Leu Leu Val Lys Ser Leu

290 295 300

Leu Thr Val Arg Ala Glu Thr Glu Thr Pro Val Ser Ala Val Ser Ile

305 310 315 320

Arg Arg Ile Pro Arg Val Pro Ala Asp Met Pro Leu Tyr Asp Ile Leu

325 330 335

Asn Glu Phe Gln Lys Gly Ser Ser His Met Ala Ala Val Val Lys Ala

340 345 350

Lys Pro Lys Ile Val Pro Leu Pro Asp Lys Thr Glu Pro Asn Arg Glu

355 360 365

Val Ser Gly Ala Pro Gln Leu Thr Ala Pro Leu Leu Ser Asn Asn Glu

370 375 380

Glu Arg Val Glu Ser Leu Val Val Asp Ile Glu Lys Pro Gln Ser Arg

385 390 395 400

Gln Val Asn Gly Asn Lys Pro Cys Ser Met Gln Gln Asn Glu Met Pro

405 410 415

Tyr Ala Met Ser Arg Ser Ser Glu Asp Ile Asp Asp Gly Glu Val Ile

420 425 430

Gly Ile Ile Thr Leu Glu Asp Val Phe Glu Glu Leu Leu Gln Glu Glu

435 440 445

Ile Val Asp Glu Thr Asp Glu Tyr Val Asp Val His Lys Arg Ile Arg

450 455 460

Val Ala Ala Ala Ala Ala Ala Ser Ser Val Ala Arg Ala Pro Ser Ile

465 470 475 480

Arg Arg Leu Thr Gly Gln Lys Gly Thr Gly Thr Gln Asn Arg Gln Gly

485 490 495

Gln Pro Thr Gly Ile Leu Lys Lys Pro Thr Glu Gly Asp Ser Asn Pro

500 505 510

Ser Lys His Gln Val Asn Leu Val Glu Pro Leu Leu Glu Asn Lys Arg

515 520 525

<210> 4

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

gccactagcc aagtgacata t 21

<210> 5

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

gaacattata tttgatgcca tgc 23

<210> 6

<211> 19

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

caacgatcca cgggccaac 19

<210> 7

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

acgacggacg tgcgtgcgtg 20

<210> 8

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

ggctgtatag acgttctttg c 21

<210> 9

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

ggctgacaca atcttgatga cg 22

<210> 10

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

gtcgacattg tcgtggtcgt c 21

<210> 11

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

ggcgatgtca ttgcgccaac 20

<210> 12

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

gcccatgcgt tgcaactgga t 21

<210> 13

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

caagcacagt aaaagacctc g 21

<210> 14

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

cttactatca actccgagct g 21

<210> 15

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

catatggcag catgtgtaca gg 22

<210> 16

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

ggagtcatct tgtccggttt 20

<210> 17

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 17

cctgctttgg aatggtggta 20

<210> 18

<211> 37

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 18

acatgattac gaattcaact cacgcatgca tgctgct 37

<210> 19

<211> 40

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 19

tcgggtacag actagtcgag atagatacgc tagctattcc 40

<210> 20

<211> 36

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 20

cgaacgatag ccatggatgg ctgcggcggt ggggag 36

<210> 21

<211> 40

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 21

ggtaggatcc actagtcctc ttattttcca aaagaggttc 40

<210> 22

<211> 19

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 22

aaccagctga ggcccaaga 19

<210> 23

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 23

acgattgatt taaccagtcc atga 24

<210> 24

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 24

gaggagatag tggatgaaac tg 22

<210> 25

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 25

gatggattgg aatcaccttc ag 22

<210> 26

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 26

ggtggggagg gcgagagatg gct 23

<210> 27

<211> 27

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 27

gacaataggg agtctaatgt ttcaatc 27

<210> 28

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 28

gaatctggag aatcgttggc aatc 24

<210> 29

<211> 28

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 29

ctatattcat gcatacatct aaagttac 28

<210> 30

<211> 26

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 30

gaggaggcta tgacacctat tgagtc 26

<210> 31

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 31

caaggatttt gccgattgct tc 22

Claims (7)

1. The application of the encoding gene of the rice DPS1 protein or the biological material containing the encoding gene in the improvement of the panicle type of the rice DPS1 mutant; the rice DPS1 mutant has single base mutation from G to A at the splicing site of the 11 th exon and the 11 th intron of the DPS1 gene, namely, the 7427 th nucleotide of SEQ ID No. 1; the amino acid sequence of the rice DPS1 protein is shown as SEQ ID No. 3; the nucleotide sequence of the coding gene is shown as SEQ ID No. 2; the biological material is a carrier, engineering bacteria or an expression box; the spike type is the number of grains per spike.

2. The coding gene of the rice DPS1 protein or the application of the biological material containing the coding gene in improving the seed setting rate and the spike grain number of the rice DPS1 mutant or improving the yield of the rice DPS1 mutant; the rice DPS1 mutant has single base mutation from G to A at the splicing site of the 11 th exon and the 11 th intron of the DPS1 gene, namely, the 7427 th nucleotide of SEQ ID No. 1; the amino acid sequence of the rice DPS1 protein is shown as SEQ ID No. 3; the nucleotide sequence of the coding gene is shown as SEQ ID No. 2; the biological material is a carrier, engineering bacteria or an expression box.

3. The application of the coding gene of the rice DPS1 protein or the biological material containing the coding gene in restoring the spike fertility at the top of the DPS1 mutant; the rice DPS1 mutant has single base mutation from G to A at the splicing site of the 11 th exon and the 11 th intron of the DPS1 gene, namely, the 7427 th nucleotide of SEQ ID No. 1; the amino acid sequence of the rice DPS1 protein is shown as SEQ ID No. 3; the nucleotide sequence of the coding gene is shown as SEQ ID No. 2; the biological material is a carrier, engineering bacteria or an expression box.

4. The application of the coding gene of the rice DPS1 protein or the biological material containing the coding gene in improving the activity of enzymes SOD and CAT of the spikelet antioxidant system at the top of the rice DPS1 mutant spike or improving the programmed cell death of the spikelet of the rice DPS1 mutant; the rice DPS1 mutant has single base mutation from G to A at the splicing site of the 11 th exon and the 11 th intron of the DPS1 gene, namely, the 7427 th nucleotide of SEQ ID No. 1; the amino acid sequence of the rice DPS1 protein is shown as SEQ ID No. 3; the nucleotide sequence of the coding gene is shown as SEQ ID No. 2; the biological material is a carrier, engineering bacteria or an expression box.

5. The use of a gene encoding rice DPS1 protein or the biomaterial of claim 3 for reducing the top spikelet degeneration of a rice DPS1 mutant; the rice DPS1 mutant has single base mutation from G to A at the splicing site of the 11 th exon and the 11 th intron of the DPS1 gene, namely, the 7427 th nucleotide of SEQ ID No. 1; the amino acid sequence of the rice DPS1 protein is shown as SEQ ID No. 3; the nucleotide sequence of the coding gene is shown as SEQ ID No. 2; the biological material is a carrier, engineering bacteria or an expression box.

6. The SNP molecular marker related to the rice top spikelet degeneration trait is characterized in that the nucleotide sequence of the SNP molecular marker is shown as SEQ ID No.1, wherein the 7427 th nucleotide is G/A.

7. The use of the molecular marker of claim 6 for elimination of top spikelet degenerated rice.

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