CN102851299B - Application of OsDLS1 gene in regulating and controlling rice leaf senescence and heading stage - Google Patents

Abstract

The invention relates to an application of an OsDLS1 gene which is a family member of HD-ZipIII in regulating and controlling rice leaf senescence and heading stage, wherein the nucleotide sequence of the OsDLS1 gene is represented by SEQ ID NO.1, protein with 855 amino acids coded by the OsDLS1 has the sequence represented by SEQ ID No.2, and the mRNA sequence of the OsDLS1 gene is represented by SEQ ID No.3. According to the nucleotide sequence of the OsDLS1 gene disclosed herein, the rice yield can be further raised, the loss of the rice yield caused by poor environment is reduced, a design scheme can be provided for molecular breeding of rice, and an important meaning of ensuring food safety is achieved.

Description

The OsDLS1 gene is in the application aspect adjusting and controlling rice leaf senile and heading stage

This research obtains the subsidy of project of national nature science fund project (No. 31171515) and Tianjin application foundation and cutting edge technology research plan main project (No. 11JCZDJC17900).

Technical field

The invention belongs to the paddy rice integral body situation of growing and improve technical field, relate to the breeding for paddy rice, the research work of breed improvement, the HD-Zip III family member OsDLS1 gene of more specifically saying so, and this gene is in the application aspect adjusting and controlling rice leaf senile and heading stage.

Background technology

China is maximum in the world rice producing country and country of consumption, and rice yield is directly connected to China's grain security, and therefore improving rice yield has very important strategic meaning to guaranteeing China's agricultural sustainable and stable development and social stability.China has obtained important breakthrough in the breeding of hybrid rice super high-yielding, hybrid rice output increases substantially.But the especially easy early ageing of inter-subspecies hybrid paddy rice of hybrid rice makes seed circularity poor (Yuan Longping 1990).The Plant Senescence is the coefficient result of external environment and idiotype network, is the result of gene program expression in the plant.Under the influence of bad external environment, aging early takes place in rice leaf regular meeting, and the too early flavescence of blade is withered, thereby makes the rice nutrition poor growth, causes its reproductive organ grouting not to be full of, and unfilled gtains increases, and has influenced the output of paddy rice greatly.Because rice leaf is one of topmost green organs of paddy rice, be that paddy rice carries out the topmost organ of photosynthesis generation nutritive substance, it is disorderly that rice leaf early ageing often causes that source, storehouse relation transforms, and generation and the transportation of nutritive substance are obstructed, thereby cause the paddy rice underproduction.In addition, rice leaf early ageing causes the too early out of season disintegration of cellularstructure, and endomembrane system destroys, and macromolecular substance degraded etc. makes whole plant decline.Therefore, from paddy rice, separate important function gene in the old and feeble regulatory pathway, further investigate its function, understand fully its regulatory pathway in the rice leaf aging course, the mechanism of research rice leaf aging, development and improve the old and feeble regulated and control network of rice leaf, be conducive to solve the early ageing problem of late growth stage of rice, for further raising rice yield, the loss of the rice yield that minimizing causes because of poor environment, for the molecular breeding of paddy rice provides design, guarantee that grain security has great importance.

In addition, be the Main Agronomic Characters of paddy rice the heading stage of paddy rice, China is from area, low latitude Hainan (~ 22 ° of N, the short day torrid areas) to area, Heilungkiang, high latitude northeast (~ 45 ° of N, the long day cold district) there is cultivated rice to distribute, even in Heihe (~ 50 ° of N) also has a small amount of distribution (Xu Wenxia, 1999).The adaptability at the distribution that cultivated rice is so broad and the Main Agronomic Characters-heading stage of paddy rice is closely related, be that paddy rice is in long-term cultivation domestication process, under the artificial selection condition, some adapt to the mutational variety of local environment condition and preserve, make it under local condition, to ear and bloom, and applied.Therefore can promote furnishing fresh evidence and vision to the adaptive molecular mechanism research of rice ear sprouting period for the adaptability in the domestication history of paddy rice and paddy rice future, provide scientific basis for breeding, the breed improvement of paddy rice.

Plant leaf is most important to growth and development of plant, and it can be by photosynthesis with luminous energy and CO ₂Change into the utilizable organic compound of plant, for growing of itself and other histoorgan of plant.In recent years, rice leaf is crossed presenility and is caused the paddy rice underproduction to cause domestic attention, and some investigators study from the angle of plant physiology, for example the Ca of lower concentration ²⁺Can improve chlorophyll content and protein content in the blade, improve peroxidase activity, delay the senescence process (Zhu Cheng etc. 2002) of blade; In addition, by reducing source, storehouse ratio, can slow down the decline (Duan Jun etc. 1997) of protein content and chlorophyll content in the hybrid rice blade.Though the research of existing many decades aspect Physiology and biochemistry both at home and abroad is still unclear and perfect for the gene regulatory network of plant leaf aging.

Under the normal growth condition, the plant leaf aging is the process of an active.Plant leaf is old and feeble mainly by the leaf age information Control, when leaf development ripe and reach certain leaf age after, blade cell starts old and feeble program automatically.Though the aging of plant leaf mainly is that it still is subjected to the influence of external environment by an active process of inherent Gene Handling.The extraneous factor of inducing plant aging mainly comprises some biologies and abiotic stress.Biotic factor mainly shows infecting of disease and pest, and after plant was subjected to disease insect bite evil, on the one hand, plant can start self-protective mechanism, shows a kind of allergy, thus the inducing plant aging.In fact, this allergy that produces of plant also is a kind of programmed cell death (Van Doorn, 2005).The abiotic factor of inducing plant leaf senile comprises physical abuse, arid, the high heat of high temperature, reaches oxidative stress etc.These abiotic stress normal with different hormone approach (regulatory pathways such as ethene, ABA, JA) interact the aging of plant organ, for example in the Sunflower Receptacle, HD-Zip transcription factor family member HAHB4Transcription factor expression amount under sick worm erosion, physical abuse and water stress condition rises rapidly.In addition, when handling with ethene or methyl jasmonate MeJA, HAHB4Transcriptional level also increase sharply.Overexpression HAHB4The transfer-gen plant of gene is before and after handling with mechanical wounding, and the content of JA and ethene has all increased, and shows HAHB4The resultant quantity of positive regulation JA and ethene, thereby the aging (Manavella of positive regulation plant leaf Et al. 2008).

The plant leaf aging at first begins to spread to centre and the base portion of leaf from blade tip or leaf margin, and blade flavescence gradually is withered to the whole blade death that comes off.In the process of leaf senile, obvious variation is exactly chlorophyllous degraded.In Arabidopis thaliana and paddy rice, identified the functional gene of several controlling chlorophyll degradeds recently by several green mutant that stagnate.Wool grass undergraduate course study group of Fudan University utilizes the model plant Arabidopis thaliana to identify a key gene regulating and control chlorophyll degradation in the green plant organ senescence process AtNYE1(Ren Et al.2007), disclosed plant in senescence process, how the photosynthetic essential substance chlorophyll of plant leaf is by procedural degraded.Paddy rice NYC1Gene is from the stagnant green mutant of paddy rice Nyc1Middle clone obtains, NYC1Be the integral part of LHCPII (light-harvesting complex II) among the photosystem II (PSII), be responsible for chlorophyllous degraded (Kusaba Et al. 2007). Nyc1In the mutant, in plant leaf senescence phase, chlorophyll can not correctly be degraded, thereby makes the mutant blade still show green in old and feeble period.(Sato such as Sato Et al. 2009) reported one similar Nyc1Mutant Nol, this mutation type surface with Nyc1Similar, chlorophyll can be kept the long period in the ripening stage, and the chloroplast grana laminated structure also keeps stable in ripe senescence phase. Nyc1nolDouble-mutant can suppress chlorophyllous degraded lastingly.Another paddy rice green mutant that stagnates SgrBlade is compared with the wild-type plant, can keep the green of longer time in the old and feeble later stage, has postponed leaf senile.Cross expression SGRObviously accelerated the degraded of leaf chlorophyll, blade presents pale brown color table type, shows SGRPositive regulation rice leaf aging (Park Et al.2007).(Jiang such as Jiang Et al.2007) also reported one from the stagnant green mutant of paddy rice SgrMiddle cloned genes SGR, the chloroplast transit peptides of a deduction of this genes encoding, the low abundance of performance composing type is expressed under the normal growth condition, but dark induce or the ABA treatment condition under, expression level increases, and expression level descends under phytokinin CTK treatment condition.This gene of overexpression has accelerated the aging of rice leaf, shows this SGRCan the aging of positive regulation rice leaf.

Plant senescence often is subjected to phytohormone Regulation, and plant hormone all worked in each period of leaf senile, and aging has the positive regulation effect to ethene for plant leaf, increases ethylene content and can accelerate the plant leaf aging.The Guo Hong of Peking University the has defended study group's systematic research signal transduction pathway of plant hormone ethene regulation and control plant leaf aging, and disclose different plant hormones are regulated and control plant leaf jointly by interaction aging (Chen Etal. 2009; Zhong Et al. 2009,2010).Jing etc. have reported the regulatory pathway that old and feeble approach that ethylene signaling and leaf age are relied on combines, and utilize Arabidopis thaliana Old1( Onset of leaf death 1) mutant clone gone out the key gene in this regulatory pathway OLD1Give Old1Mutant applies exogenous ethylene then can accelerate early stage leaf senile, shows OLD1Effect (the Jing of negative regulation ethene in leaf age dependence leaf senile Et al. 2005).(Cytokinin CTK) has important effect for delaying The Plant Senescence to phytokinin.Phytokinin is mainly synthetic at root, transfers to the plant other parts by xylem and works.(Kim such as Kim Et al. 2006) utilize Arabidopis thaliana function gain mutation body Ore12-1The clone has obtained the CTK acceptor gene AHK3, find that it plays a crucial role in the leaf senile regulation and control of CTK mediation.

Some receptor kinases are also related to the plant leaf aging by evaluation, and Wang Ningning study group of Nankai University has identified a soybean receptor kinase gene GmSARK, cross and express the senescence process that this gene has accelerated soybean leaves, show that this gene relates to the old and feeble regulation and control of soybean leaves (Li Et al.2006).These receptor kinases are targets of WARK transcription factor, regulate and control leaf senile (Robatzek jointly with regulation and control WARK transcription factor interaction Et al.2002).In addition, the plant leaf aging is subjected to the adjusting of sugared concentration in inducing of optical condition and the blade, Arabidopis thaliana hexokinase gene HXK1Be a gene that typically is subjected to the aging of intensity of illumination regulation and control plant leaf, this gene is from the insensitive mutant of glucose Gin2Middle clone obtains, and when the external source glucose concn reached 6%, the wild-type vine growth and development was stagnated, and Gin2Still can normal growth.Under the normal illumination condition, wild-type and mutant blade performance normal growth, yet, under high intensity of illumination is handled, the rapid old and feeble flavescence of wild-type plant leaf, and Gin2Though the mutant plant has reduced increment, its leaf senile postpones, leaf look dark green (Moore Et al.2003).Show HXK1Regulate and control The Plant Senescence by integrating exterior illumination signal and inherent gene signal.

Summary of the invention

The object of the present invention is to provide HD-Zip III family member OsDLS1Gene and application thereof.

Above-mentioned purpose of the present invention is to be achieved by following method:

HD-Zip III family member OsDLS1The nucleotides sequence of gene is classified as shown in the SEQ ID No.1, OsDLS1855 the amino acid whose protein sequences of encoding are shown in the SEQ ID No.2.

HD-Zip III family member of the present invention OsDLS1The mRNA sequence of gene is shown in the SEQ ID No.3.

The present invention further discloses HD-Zip III family member OsDLS1The application aspect preparation adjusting and controlling rice leaf senile and rice ear sprouting period of gene and protein.

Experimental result of the present invention shows:

(1) HD-Zip III family member OsDLS1Gene can the adjusting and controlling rice blade aging, after the forfeiture of OsDLS1 gene function or descending, accelerated the aging of rice leaf, when OsDLS1Delayed the aging of rice leaf after the gene function rise (cross and express).

(2) OsDLS1Gene can promote the heading of paddy rice under the long day condition, and after the forfeiture of OsDLS1 gene function or descending, paddy rice is postponed the heading stage under the long day condition.

Description of drawings:

Fig. 1 OsDLS1 transcription factor structural domain and subfamily member thereof; A:OsDLS1 transcription factor structural domain wherein; B: HD-ZIP III subfamily member kinship in the paddy rice;

The phenotype of Fig. 2 RNAi transfer-gen plant; The phenotype of A:RNAi transfer-gen plant wherein.The at first blade tip flavescence of RNAi rotaring gene plant blade, rust staining appears in the blade middle and lower part, and rust staining extends to whole blade afterwards, finally makes the blade withered death.B:RNAi transfer-gen plant destination gene expression detects, and CK is the empty carrier transfer-gen plant.C: OsDLS1The expression of homologous gene in the RNAi transfer-gen plant;

Fig. 3 crosses the phenotype of express transgenic plant; A wherein: the phenotype of crossing the express transgenic plant.When the wild-type plant leaf begins to dry up, cross the express transgenic plant leaf and also be in complete green, do not show withered.B: cross express transgenic plant destination gene expression and detect, WT is that to spend 11, OV1 ~ OV5 in the wild-type be the express transgenic plant;

Fig. 4 OsDLS1Expression in different tissues; A wherein: OsDLS1Expression under leaf, the second internode stem, fringe in stem, young fringe, ripe floral organ and the young tender meristematic tissue.B: OsDLS1Expression at the leaf different development stage.

Embodiment:

In order to explain enforcement of the present invention more fully, provide HD-Zip III family member OsDLS1 the preparation embodiment of gene.These embodiments only are to explain rather than limit the scope of the invention.Wherein used reagent all has commercially available.

Embodiment 1

1 vegetable material: spend 11(to have commercially available in the japonica rice variety) (Oryza sativa L. ssp. Japonica) is used for this experiment.

Method

2.1 the structure of RNAi carrier

We have made up two RNAi carriers, and an interference region section is positioned at 5 '-UTR district, and another interference region section is positioned at 3 '-UTR district.Interfere sections to obtain from the oryza sativa genomic dna amplification for two that are used for structure RNAi carrier, the primer sequence is: RNAi-3 ' UTR:IHD1:5 '-CACCATCCATGGTTTCCAG-3 ' and IHD2:5 '-GGTAACTCAATGCCGATTGC-3 '.RNAi-5 ' UTR:IHD3:5 '-CACCGGTCGAGAGA GAAAGT-3 ' and IHD4:5 '-TCCTCCTCGTCGCTGCTTGT-3 '.The fragment pENTR that amplification is good ^TMDirectional TOPO Cloning Kits (Invitrogen, USA) according to the program construction on the specification sheets in purpose carrier pANDA35HK, cut and PCR method is identified in the method rice transformation material that correct clone mediates with agricultural bar and spent 11 (Hiei et al. 1994) by enzyme.The pcr amplification condition is: 1 min/95 ℃; 30 cycles (30 sec/94 ℃, 30 sec/59 ℃, 30 sec/72 ℃); 5 min/72 ℃.

Reaction system following (cumulative volume 20 μ L):

DNA 10－30ng

10 * Buffer (contains 20 mM Mg ²⁺) 2 μ L

dNTP（2.5mM） 　　0.2mM

PrimerF (IHD1/IHD3) 0.2μM

PrimerR (IHD2/IHD4) 0.2μM

ExTaq enzyme (TaKaRa) 1 U

DdH ₂O mends to 20 μ L

2.2 cross the structure of expression vector

Have complete ORF's OsDLS1The cDAN of gene obtains by the RT-PCR method from the rice seedling of 15 days sizes, introduces in design of primers SmaI and SacTwo restriction enzyme sites of I are used for corresponding restriction enzyme site with carrier and are connected.The primer sequence of amplification is: OSHDF:5 '-ATGA CCCGGGCGAGTTCTTGCTGGGTTGTTG-3 ' and OSHDR:5 '-TTGC GAGCTCACGGTGGTGGTATTCAGGTCT-3 '.Connect back transformed into escherichia coli DH5 α (TaKaRa, Dalian), cut by enzyme and identify and after PCR identified, positive colony carried out sequencing analysis that the correct carrier that checks order is with spending 11 in the agrobacterium mediation method conversion.The pcr amplification condition is: 1 min/95 ℃; 35 cycles (30 sec/95 ℃, 30 sec/63 ℃, 3min/72 ℃); 5 min/72 ℃.

Reaction system following (cumulative volume 20 μ L):

cDNA　　　　　　　 1μL

10 * Buffer (contains 20 mM Mg ²⁺) 2 μ L

dNTP（2.5mM） 　　 0.2　mM

OSHDF 0.2μM

OSHDR 0.2μM

Phusion Taq（NEB） 1U

DdH ₂O mends to 20 μ L

2.3 OsDLS1The expression of gene in the different tissues organ

In order to analyze OsDLS1The expression of gene in paddy rice different tissues organ, rice leaf, stem, young fringe, ripe floral organ and young tender meristematic tissue are collected, (Invitrogen, (Invitrogen USA) handled digested genomic dna to total RNA with DNase I after USA) reagent extracted with TRIzol.(China) cDNA is synthesized in reverse transcription to total RNA of 1 μ g for TaKaRa, Dalian with the M-MLV ThermoScript II.Synthetic cDNA uses OsDLS1The special primer of gene carries out PCR reaction amplification, with paddy rice OsActin1Doing confidential reference items analyzes relatively OsDLS1Expression of gene.Genetic expression special primer sequence is:

FDF:5 '-CACCACGCGACTTTTGGACT-3 ' and FDR:5 '-GGAGATTCATAGAGCGGTCG-3 '.Amplification program is 1 min/95 ℃; 30 cycles (30 sec/94 ℃, 30 sec/59 ℃, 30 sec/72 ℃); 5 min/72 ℃.

2.4 OsDLS1Expression analysis in RNAi and mistake express transgenic plant

In order to detect RNAi and to cross the interference effect of express transgenic plant and cross expression effect, total RNA extracts the back from the blade of the transfer-gen plant of 40 days sizes and contrast thereof (Invitrogen USA) handles digested genomic dna with DNase I.(China) cDNA is synthesized in reverse transcription to total RNA of 1 μ g for TaKaRa, Dalian with the M-MLV ThermoScript II.The cDNA that counter-rotating records is used for RT-PCR or Real-time pcr template.(China) test kit description operation is at Stratagene Mx3000P for Tiangen, Beijing according to SYBR Green PCR master mix for Real-time PCR ^TM(STRATAGENE USA) carries out amplified reaction on the PCR instrument to Thermal System.The primer sequence is with 2.3 used sequences.Each reaction is carried out 3 times and is repeated, and uses paddy rice OsActin1As the confidential reference items relative quantitative assay, 2 of Livak description is adopted in data analysis ^{-△ △ Ct} Method is carried out (Livak et al. 2001).Real-time pcr amplification condition is: 2 min/95 ℃; 40 cycles (95 ℃ of 20 sec/, 59 ℃ of 30 sec/, 68 ℃ of 30 sec/).

Interpretation of result

3.1 OsDLS1Gene structure and subfamily analysis

OsDLS1855 the amino acid whose protein of encoding contain 4 structural domains, wherein the HD-ZIP structural domain that constitutes of Homeobox and bZIP play an important role in this transcription factor (Figure 1A).Special dna sequence dna can with HD-ZIP in conjunction with the territory in conjunction with and other gene is played regulating and controlling effect.In the paddy rice, HD-Zip III subfamily has 5 members, their kinship as shown in Figure 1B, wherein OsDLS1With OsHOX32Sibship is nearest, and amino acid sequence identity is up to 87%. OsDLS1Also very high with the HD-Zip III subfamily member's of other plant homology, with the amino acid identity of ATHB9, the ATHB14 of known Arabidopis thaliana and ATHB15 also all more than 65%.The aminoacid sequence that shows this transcription factor subfamily is very conservative.

OsDLS1The RNAi transfer-gen plant obtains and phenotype analytical

In order to explore the function of this transcription factor, we adopt the reverse genetics method to make up RNAi and cross expression vector, spend in 11 during it is imported to.Two RNAi carriers have obtained 108 strain T0 altogether for transgenic positive plant (61 strain RNAi5 '-UTR, 47 strain RNAi3 '-UTR), but do not see obvious phenotype.20 RNAi5 ' of picked at random-UTR T0 obtains T1 for plant for plant and 12 RNAi3 '-UTR T0 for the plant sowing, there are 8 strain systems that phenotype is arranged in 20 RNAi5 '-UTR T1 strain systems, show rust staining on the beginning plant leaf, patch expansion is afterwards drawn out to whole blade, finally makes blade withered death (Fig. 2 A).12 RNAi3 '-UTR T1 are for there being 2 strain systems that phenotype is arranged in the strain system, and are slightly different with RNAi5 '-UTR phenotype, at first blade tip flavescence of plant leaf, and rust staining appears in the blade middle and lower part, and rust staining extends to whole blade afterwards, finally makes blade withered death (Fig. 2 A).The sampling of listing of phenotype plant will be arranged, extract RNA, with RT-PCR method testing goal expression of gene, find in the RNAi transfer-gen plant, OsDLS1Expression obviously reduce compared with the control, show that RNAi has disturbed really OsDLS1Expression of gene (Fig. 2 B).Individual plant is received kind of the T2 that the plantation acquisition is isozygotied and is used for follow-up study for transfer-gen plant.

T1 is (long day area, ~ 39 ° of N) and Hainan (~ 22 ° of N) plantation respectively in Tianjin in 2011 for the RNAi transfer-gen plant, finds the heading stage of statistics RNAi transfer-gen plant, OsDLS1The RNAi transfer-gen plant was postponed 35 days than contrast at the heading stage of Tianjin plantation, and postponed 6 days than contrast at the heading stage of Hainan plantation.Because the photoperiod of two regional field plantings is all different with temperature, we plant (LD:14h illumination, 10 h dark under the growth cabinet control condition of laboratory; SD:10h illumination, 14 h dark; Temperature all is 28 ℃) RNAi transfer-gen plant and contrast, the statistics heading date is found transfer-gen plant and contrast heading substantially simultaneously under the short day condition, but has postponed more than 40 days at the heading stage of transfer-gen plant under the long day condition.This result shows that this transcription factor can promote the heading of paddy rice under the long day condition.

Interfered other member of HD-ZIP III subfamily in order to detect constructed RNAi carrier, we with RT-PCR detected with OsDLS1The expression of several genes in the RNAi transfer-gen plant of homology, the result shows that RNAi3 ' UTR carrier is very special, can not interfere other HD-ZIP III subfamily member, but RNAi5 ' UTR carrier can be interfered OsHOX32Gene, but can not interfere other 4 members (Fig. 2 C).This may also be the slightly different reason of transfer-gen plant phenotype that the RNAi carrier of two structures obtains.

OsDLS1Crossing the express transgenic plant obtains and phenotype analytical

Cross expression vector and obtain 68 strain T0 altogether for plant, wherein 42 strains are the transgenic positive plant.6 strain blades performance delay senility is arranged in the positive transfer-gen plant, and fringe and floral organ performance deformity (Fig. 3 A).All the other plant do not have obvious phenotype difference.Individual plant is listed to take a sample and is extracted RNA, with RT-PCR method testing goal expression of gene, crosses the express transgenic plant OsDLS1The expression of expression of gene obviously strengthens (Fig. 3 B).

OsDLS1Expression analysis in different tissues

In order to detect OsDLS1Expression in the paddy rice different tissues is collected wild-type different time and different tissues material, isolation of RNA, and with the spatial and temporal expression characteristic of RT-PCR analysis purposes gene in the paddy rice different tissues, the result shows after the reverse transcription OsDLS1Expression is all arranged in the paddy rice different tissues, but higher at children tender meristematic tissue and the expression amount in the young fringe, expression amount lower (Fig. 4 A) in the floral organ of maturation.Whole growing OsDLS1All express.In order to detect OsDLS1In the expression of rice leaf different development stage, analyzed in young tender blade (15 days seedling leaves), mature leaf (60 days plant sword-like leaves) and the old and feeble blade (blade begins flavescence) with real time quantitative PCR method OsDLS1Expression, the result shows that expression amount is higher in the tender blade of children, and the expression amount in old and feeble blade lower (Fig. 4 B), shows that this transcription factor expression amount in the tender tissue of children is higher, along with the old and feeble expression amount of tissue descends.

OsDLS1Gene order (shown in the SEQ ID No.1):

AAATAAGAAACAACGAGACTCTTGATGTACCAAGCCGAGTCTGAAGAAGAGAAAAGAGAGAGAGTTCAGAAAAAGGAAACGACCCTATTCTGATCAGAAAAATTCAATCTTTCTCTCTCTTTCTCTCAGAAAAGGTCGAGAGAGAAAGTGGATGTCTTCATGTGAAGACCAGAGAGAAGACGAGAGGCGATCGCAGTGGAGTGGAGTGAAGTACTAGCAGTATTAAAACCAGCAGCAAGCGAGAGAGAGGCCACCAACCACTTCCTCCTCCAAGAAACCACCCCCACCACGAGCACGAGCACGAGCAACCATGGCGGTGGCAGAGGGAGCTGCGACTGCGAGCTACGGGCCATGACTGCGGCGAGTTCTTGCTGGGTTGTTGTTGTACAAGCAGCGACGAGGAGGAGGGGAGCAGAGGAGGAGGAGGCGGAGGGATGGCAGCGGCGGCGGTGGGGGGAAGGGGGGAGAGGCTGTCGTCCTCCTCCCCGACGGCGGCGGCGCCGCAGGTGGACGCCGGGAAGTACGTCCGGTACACGCCCGAGCAGGTGGAGGCGCTGGAGCGGGTGTACACCGAGTGCCCCAAGCCCAGCTCACTCCGCCGCCAGCAGCTCATCCGGGAGTGCCCCATCCTCAGCAACATCGAGCCCAAGCAGATCAAGGTCTGGTTTCAGAATCGCAGGTCAGTCTCATCTCCTTCTCTCTCTTCGATCTGTTTCTGCTCGGGTTTGTTCAGCAGCCATGGCGTGTGTTCTTCGGTTTCTTTTCTTGGTTTTGTGTAATGTGATGAGATTGATTGCTTTGTTGTTTGCAGTTTTTTTTCATGTTCTTGTGGGAATGGGGGATCCTTTGTGAGGAAGATTTGGGTTCTTGAGATGAGTTTTTTTTTTCTGTTTGATTTGTAGATGTCTGTAACTTTTTGGGTGTTGATTTTAGGGCTTGCAATGTGTTGGATTGGTGACGAGAAGGTTCATTTCTGGGATTTGCATCCTATATGATGGGTTTTGTTTTTTTAGTTCGTCTTGTGCAATAATGAAGCGCAATCTGTTTGCTTGAAATTTATCAGGATATTTTCTTTTATAAAACTTTGCGGCAATGATTTGAGTTCCTGTGCCCACACTGTTCTTCGCTGCATTATTCTGCTGGGTTGTGTGGTGGGTAGGGAGCGAAATGGCAGCATCATTCGTTGGTTTCCTCTTGGATTTTGTGGTGATGTGAATAGCCAAATATCCTGGATCTCTGTACAGACTTTACAGTTAATTTTGTTTGGTTTAAATTGTAAATATATGCTCTCTGATTGTTTTGTTGCTTGTGGTGACCAATGATTGGTTGGAGCAAAATTGCCTTAGGTCATTTCTTGTTTCCTGTGCTTGCCATTTTAAGGGCCATCTTTTTAACCCTGTGTTGTGGGCTGTGGTAGTGTTCATGTTTTCTTGCTATAGGCCTCCATAAGCATCACCTCCTGGTTATCATCTCGCTTTCTTTTTTCAGTGGATCACAAGTAGGGCAATAGTTTTGTCACTGGTGGTGTCACTGTGGAAGAATGTGAAGGGCAACATTTTGAGCAAGTTTGGTATATACTATTACTGTACTCATGAAGGCCAAATATAAATCAAGAATCCTTTTTAGGTAGAGGATGCTCTTGTTTAGAGTTGAGTTCACAGATGCAGACAGAAACATCACTGTCAATTAGGCAATGATTTTCACATATGGAATACTAACTGTTATTGCATAGTATATAGTCATTTGTCTCTTTTGCAAATGACCTTTTGCTTGTTCCTGTTCAATGATCCCCCTTTTAACCTTGGGGAATATTTGCTAAAATGCCTTTGCCTAACTTCCTTTGTGCTCATTTGGGCTGTGCCAATCAGTATCCATTAACAAGTATTGGAATCTTGTGTATTAGAAGACACCATCCTTTAGATAGTTTATTCATGTACTTTTGCCACTGAAATTGTGCCTTCCTTGTAATTGGCAGATGCCGGGAGAAGCAGCGCAAGGAGGCCTCCCGGCTGCAAACTGTGAACCGGAAGCTGAATGCGATGAACAAGCTGTTGATGGAGGAAAATGACCGGCTGCAGAAGCAGGTGTCCCGTCTCGTCTACGAGAACGGCTACATGCGGACTCAACTGCACAATGTGAGCTCCCTGTGCTTACAGGCTTTTGTTCTTACAATTGCTTCATATTTTGATCATCCTGTTTTCTTAATCAGTAATGTTTGTATGTACTTTGTAGCCTTCTGCAGCCACCACAGATACAAGCTGTGAGTCTGTGGTGACAAGTGGTCAGCACCATCAACAGCAAAACCCAGCAGTTCTGCATCCGCAAAGGGATGCGAACAACCCAGCAGGGTATCCACAAAACCTGGTCTATTTTCTTCTTTTGTTAGGCCGTGCTTTTTTGTATGAAAGATTGCTGAGATGATTTTATTTGCGCAGTCTTCTCGCTATTGCTGAGGAGACCTTGGCAGAGTTCATGTCGAAGGCGACAGGAACTGCTGTCGAATGGGTGCAAATGGTTGGGATGAAGGTAATGTATTCGTTGTACAAGAAAACTGTGCAGTATGTTGATTTTCTCTTTCCATTCATGTTTTTTTTGTTCAAATCAACTGAAATGAAAATTTGATTACAGCCTGGTCCGGATTCCATTGGAATCATCGCTGTTTCGCACAATTGCAGTGGCGTAGCAGCACGAGCTTGCGGCCTGGTGAGCCTTGAGCCCACAAAGGTATGATTTGTTTTCCTTGATATTGTGTTACATTTCCCATTAACTGGCATGCTACAGACCCTGATAAGCTTCATTTTAGGTTGCAGAGATCCTCAAGGATCGCCCCTCTTGGTACCGTGATTGCCGATGCGTAGATATCATCCATGTTATCCCTACGGGTAACGGCGGAACCATTGAGCTAATCTACATGCAGGTAAATATACCCAATCGTGACTCATGTGTTGCATTTTTCTCAAGCATCACTAGCTACTCTTTGTTTTTGGTAATAACCATTGCCCTTTTCAATTCCTTAGACATATGCACCGACAACTTTGGCGGCACCACGCGACTTTTGGACTCTCCGCTACACAAGTGGACTTGAAGATGGGAGTCTTGTGGTATTACTTTGATCTTTAATTGCATGCTGTCAGCTCATTAAACAAAATGCTTCCTTTCATTTTCTTGTATGGTCTATTCGTCATGTTATGATCGGACTTGTTCTGTTTGGATCATACCACATCATCATGGTTTTCAACTGAAGTGTAGTTTCTGTTGTATTAAATATTGTTTGACACCATTCCTGTTCTGTAGATCTGTGAAAGGTCATTGACTCAGTCCACTGGTGGCCCATCTGGGCCTAACACTCCGAATTTTATCAGAGCCGAGGTACTTCCCAGTGGTTATCTGATTCGACCATGTGAGGGAGGTGGCTCCATGATTTACATTGTTGATCATGTTGATTTGGATGTAAGTAACAATAGTGTTTTCACATTAACTAGTGTAATATTGAATAGTTTTGAGATACATATCTTATTAATCCTGTGTATTTTACAGGCTTGGAGTGTGCCTGAGGTTCTTCGACCGCTCTATGAATCTCCAAAGATCCTTGCACAGAAGATGACCATTGCAGTATGTGCACTGTTTTATTCTGTTGTTGAAGAGCACTTGGTCACCTCATGGTGATTGTGAGTGCATAACTTTTCTTATCACCAATGACATGGTTCATTCTGTTCAGGCATTGCGACACATTAGGCAAATTGCGCATGAGTCAAGCGGTGAAATCCCCTACGGTGCTGGTCGCCAGCCTGCTGTTTTCAGGACCTTCAGTCAAAGGCTCAGCAGGTCTGTAGCTATTTGAGGGTTCCCATGTATTAGAACCAATTGGCATACTGTCATATGACACAGTTCCATGCATGTCTCATTGCTCACGCATCTGAACTCCATACTCTAATTTGCAGAGGTTTCAATGATGCTGTTAGTGGATTTCCAGATGATGGGTGGTCTTTGTTGAGCAGTGATGGTAGTGAGGACATTACAATTTCAGTAAACTCGTCTCCAAACAAGCTTGTTGGATCCCATGTCAGCCCGAACCCTCTGTTTTCTACTGTTGGAGGTGGCATCTTGTGCGCAAAGGCATCAATGCTACTTCAGGTTGACATAGGCTCTATTTAGTTGCTCAGCTAATTATAGATTGTCTCTATCTTTGTTTCACTTTGACCAAAATTGTTCCAAATCAAAGAATTAAGCTTTACAAAATGTCATTTTTTTTATAATGTAGAATGTACCTCCTGCTCTACTTGTGCGATTTCTGAGGGAGCATCGTTCTGAATGGGCTGATCCTGGTGTTGATGCTTATTCTGCTGCTTCTTTGCGAGCCAGCCCATATGCAGTTCCAGGCTTAAGGACCAGCGGGTTTATGGGCAGTCAGGTTATACTACCCCTCGCTCATACCTTAGAGCATGAAGAGGTAATGTTGTAATTGAGTCCTTGTAGGCTTGTAGCTCCTTTTCCACAATGAATGGCAAATATTAATGCATCAACCTGGCATTCTGAATTGCAGTTCTTGGAGGTTATTAGGCTCGAAGGACATGGTTTCAGCCATGATGAGGTGCTTCTATCACGGGATATGTACCTTCTACAGGTATTGTGGACCATCCGAGTCAACTATTTATTCCTTTGCTGTTGGGCTGGACAAGTGGGCTTCGGTGGGAGGCATGCTGCTGTTCTTGGCCACCGTATGGGCCTAACCTTCTGCTCAGGGTTTCTGGTGGATCAGGTCCTCCTGCGCACGAATTATTTTTTATTTTTGTTCATAATAAAAGCTGAGATGTAAAAGGTTTAAGAGTCTTAGTTTAGGTTATGGTGCTTCGTTAAGTCTAGCAGGGTGTGCTGCAGCTCGAGGACGAGCTGCTCAAAAAGGGGGGAGTAATGTCATGGGCTTTGGGCCGGGCGTCCTAAGTCCATTGGATAGGATTAGGGTTTGTTAGGATTAGATAAGGTTTGTTAGGATTAGATTAAGGAGTCCTCCATCTATATAAGGAGGGATCCTATCTCAGGTCAATTAGGCATTAGATCAAGATTTATCTTAGTGCCCATCGGCCTGCCTTCTCAGTGCGACGGAGAGCGTCGCGCCGTTTAGGTTCAGGACCGTATTCCTTGTTATTTCATCTACTATAATCCAATCTATCCCCATTCATCCCGATTCTTCTCCACATAATCCTTGCTCATGACAATTGCCTTAAGTTGTACCTGTTTGTTGCATTTTCCTGGTTCTGAGAGATTGCTTTCTATGCAGCTTTGCAGCGGGGTTGATGAGAATGCTACTAGTGCATCTGCACAGCTTGTCTTTGCTCCTATTGACGAATCTTTTGCTGATGATGCACCACTTTTACCCTCAGGATTCCGTGTGATACCACTAGATACAAAAATGGTATGACTGGTCCATAATCTTATTCCAGAGGCCTTGTCTTTCCCATGTTGGCTCACCAATTAACCGTTATGCACTCAAACCTGCAGGATGGGCCATCTGCAACACGCACGCTCGACCTTGCATCCGCCCTTGAGGTTGGACCAGGTGGAGCTTCGCGTGCTTCAGTTGAAGCATCTGGCACATGTAACAGATCAGTCCTGACCATTGCTTTCCAATTCTCATACGAGAACCACCTTCGTGAGAGCGTGGCGGCAATGGCTAGGAGTTATGTCAGAGCTGTGATGGCATCCGTGCAGAGGGTGGCTGTGGCAATAGCTCCTTCTCGACTTGGACCCCAGATCGGAATGAAGCACCCTCCAGCCTCTCCTGAGGCGCTTACACTTGCTAGCTGGATTGGTAGGAGCTATAGGTAAGTAAAATACTTATTTTTAGCAAAAGTCTATCTCAGGTTCTTCATCATCAGAACCACCTACACATTGCCACATCTGAAGTTGTGATCAATGATGTTAATGCACTAGTTATTTCCGTGGATCTTTCAGGGCTCACACCGGAGCAGATATCCGTTGGTCTGATACCGAAGATGCCGATTCTCCCCTGGCGCTCCTGTGGAAGCACAGCGACGCAATACTGTGCTGTTCTCTGAAGGTTAGTAGTGCTGCCCACTAATTTTATGCTCATTTCATCACATGCATGTTCTGGAAACCTTGCTAATGGTCAGTTTGGTTATCTACTGGCAGCCTGCTCCTATGTTCACCTTCGCCAACAATGCCGGCCTCGACATCCTTGAGACGACGCTGGTCAACCTCCAGGACATCTCGCTCGAGATGATCCTGGATGACGAGGGCCGGAAGGCGCTGTGCTCGGAGTTCCCCAAGATCATGCAGCAGGTAAAACCACCGTGTCCCTGCCTCGCGACAATCGGTGACGCATCCATCAGAGTACTACCAGACCATGAGCAATGTTCCTGATCAAGCCTGTTTGGGGTCTGTCTATCTTCTTTCAGGGTTTCACCTATCTCCCTGGAGGCGTCTGCAAGTCGAGCATGGGGCGGCAGGCATCGTATGAGCAGGCGGTGGCGTGGAAGGTTCTTAGCGACGACGACGCGCCGCACTGCCTCGCCTTCATGCTCGTCAACTGGACCTTCATGTGATCCACCATCCATGGTTTCCAGCTTCCAGCTAGTGCTTTTGCTTTTGTGCCATTGTGGCAAACTGAACAAAGACCTGAATACCACCACCGTATTGTATTAAACATATGCGTCCATACATGCATGGTGCTTGCTTTATCCTAGCTAGTGCTAGCACCTGTGTGTATGGTCTCTCCATGTTTGAAGCTTAAAATTTTAGAACCCTGGTTTTTTTTTTTCTGCTGCTGTGTGGATGAACAAGTTTCAGGACCTATGTATCTCTATCTATCTATGCTAATGCTATGAATTGTGTGATGTGTGTATGCTTTGAGCAATCGGCATTGAGTTACCTTGGTTGGTTTATGCCGTGCTGAACCATTGTTGATGCATATGTAGCTGAAGGAAGTATTTGCTGAATGTGATTCGTATTCCTCCACCGTTAAACTAAAACAAAAGCGTTTCTACCTATCAAAACCGGCTCATTATGCTCTCAGCTTAAACCATTATCCTTTGCTGATTGTGGTACATAGTTGAGGGAGCCCAAATAGACTTTTCTCAGTGCTACGTCCTGCTCACTGATCTCGGCCACGAATAGAATCGT。

OsDLS1855 the amino acid whose protein sequences of encoding are shown in the SEQ ID No.2:

MAAAAVGGRGERLSSSSPTAAAPQVDAGKYVRYTPEQVEALERVYTECPKPSSLRRQQLIRECPILSNIEPKQIKVWFQNRRCREKQRKEASRLQTVNRKLNAMNKLLMEENDRLQKQVSRLVYENGYMRTQLHNPSAATTDTSCESVVTSGQHHQQQNPAVLHPQRDANNPAGLLAIAEETLAEFMSKATGTAVEWVQMVGMKPGPDSIGIITVSHNCSGVAARACGLVSLEPTKVAEILKDRPSWYRDCRCVDIIHVIPTGNGGTIELIYMQTHAPTTLAAPRDFWTLRYTSGLEDGSLVICERSLTQSTGGPSGPNTLNFIRAEVLPSGYLIRPCEGGGSMIYIVDHVDLDAWSVPEVLRPLYESPKILAQKMTIAALRHIRQIAHESSGEIPYGAGRQPAVFRTFSQRLSRGFNDAVSGFPDDGWSLLSSDGSEDITISVNSSPNKLVGSHVSPNPLFSTVGGGILCAKASMLLQNVPPALLVRFLREHRSEWADPGVDAYSAASLRASPYAVPGLRTSGFMGSQVILPLAHTLEHEEFLEVIRLEGHGFSHDEVLLSRDMYLLQLCSGVDENATSASAQLVFAPIDESFADDAPLLPSGFRVIPLDTKMDGPSATRTLDLASALEVGPGGASRASVEASGTCNRSVLTIAFQFSYENHLRESVAAMARSYVRAVMASVQRVAVAIAPSRLGPQIGMKHPPASPEALTLASWIGRSYRAHTGADIRWSDTEDADSPLALLWKHSDAILCCSLKPAPMFTFANNAGLDILETTLVNLQDISLEMILDDEGRKALCSEFPKIMQQGFTYLPGGVCKSSMGRQASYEQAVAWKVLSDDDAPHCLAFMLVNWTFM.

OsDLS1 mRNA sequence (shown in the SEQ ID No.3):

ATGGCAGCGGCGGCGGTGGGGGGAAGGGGGGAGAGGCTGTCGTCCTCCTCCCCGACGGCGGCGGCGCCGCAGGTGGACGCCGGGAAGTACGTCCGGTACACGCCCGAGCAGGTGGAGGCGCTGGAGCGGGTGTACACCGAGTGCCCCAAGCCCAGCTCACTCCGCCGCCAGCAGCTCATCCGGGAGTGCCCCATCCTCAGCAACATCGAGCCCAAGCAGATCAAGGTCTGGTTTCAGAATCGCAGATGCCGGGAGAAGCAGCGCAAGGAGGCCTCCCGGCTGCAAACTGTGAACCGGAAGCTGAATGCGATGAACAAGCTGTTGATGGAGGAAAATGACCGGCTGCAGAAGCAGGTGTCCCGTCTCGTCTACGAGAACGGCTACATGCGGACTCAACTGCACAATCCTTCTGCAGCCACCACAGATACAAGCTGTGAGTCTGTGGTGACAAGTGGTCAGCACCATCAACAGCAAAACCCAGCAGTTCTGCATCCGCAAAGGGATGCGAACAACCCAGCAGGTCTTCTCGCTATTGCTGAGGAGACCTTGGCAGAGTTCATGTCGAAGGCGACAGGAACTGCTGTCGAATGGGTGCAAATGGTTGGGATGAAGCCTGGTCCGGATTCCATTGGAATCATCACTGTTTCGCACAATTGCAGTGGCGTAGCAGCACGAGCTTGCGGCCTGGTGAGCCTTGAGCCCACAAAGGTTGCAGAGATCCTCAAGGATCGCCCCTCTTGGTACCGTGATTGCCGATGCGTAGATATCATCCATGTTATCCCTACGGGTAACGGCGGAACCATTGAGCTAATCTACATGCAGACACATGCACCGACAACTTTGGCGGCACCACGCGACTTTTGGACTCTCCGCTACACAAGTGGACTTGAAGATGGGAGTCTTGTGATCTGTGAAAGGTCATTGACTCAGTCCACTGGTGGCCCATCTGGGCCTAACACTCTGAATTTTATCAGAGCCGAGGTACTTCCCAGTGGTTATCTGATTCGACCATGTGAGGGAGGTGGCTCCATGATTTACATTGTTGATCATGTTGATTTGGATGCTTGGAGTGTGCCTGAGGTTCTTCGACCGCTCTATGAATCTCCAAAGATCCTTGCACAGAAGATGACCATTGCAGCATTGCGACACATTAGGCAAATTGCGCATGAGTCAAGCGGTGAAATCCCCTACGGTGCTGGTCGCCAGCCCGCTGTTTTCAGGACCTTCAGTCAAAGGCTCAGCAGAGGTTTCAATGATGCTGTTAGTGGATTTCCAGATGATGGGTGGTCTTTGTTGAGCAGTGATGGTAGTGAGGACATTACAATTTCAGTAAACTCGTCTCCAAACAAGCTTGTTGGATCCCATGTCAGCCCGAACCCTCTGTTTTCTACTGTTGGAGGTGGCATCTTGTGCGCAAAGGCATCAATGCTACTTCAGAATGTACCTCCTGCTCTACTTGTGCGATTTCTGAGGGAGCATCGTTCTGAATGGGCTGATCCTGGTGTTGATGCTTATTCTGCTGCTTCTTTGCGAGCCAGCCCATATGCAGTTCCAGGCTTAAGGACCAGCGGGTTTATGGGCAGTCAGGTTATACTACCCCTCGCTCATACCTTAGAGCATGAAGAGTTCTTGGAGGTTATTAGGCTCGAAGGACATGGTTTCAGCCATGATGAGGTGCTTCTATCACGGGATATGTACCTTCTACAGCTTTGCAGCGGGGTTGATGAGAATGCTACTAGTGCATCTGCACAGCTTGTCTTTGCTCCTATTGACGAATCTTTTGCTGATGATGCACCACTTTTACCCTCAGGATTCCGTGTGATACCACTAGATACAAAAATGGATGGGCCATCTGCAACACGCACGCTCGACCTTGCATCCGCCCTTGAGGTTGGACCAGGTGGAGCTTCGCGTGCTTCAGTTGAAGCATCTGGCACATGTAACAGATCAGTCCTGACCATTGCTTTCCAATTCTCATACGAGAACCACCTTCGTGAGAGCGTGGCGGCAATGGCTAGGAGTTATGTCAGAGCTGTGATGGCATCCGTGCAGAGGGTGGCTGTGGCAATAGCTCCTTCTCGACTTGGACCCCAGATCGGAATGAAGCACCCTCCAGCCTCTCCTGAGGCGCTTACACTTGCTAGCTGGATTGGTAGGAGCTATAGGGCTCACACCGGAGCAGATATCCGTTGGTCTGATACCGAAGATGCCGATTCTCCCCTGGCGCTCCTGTGGAAGCACAGCGACGCAATACTGTGCTGTTCTCTGAAGCCTGCTCCTATGTTCACCTTCGCCAACAATGCCGGCCTCGACATCCTTGAGACGACGCTGGTCAACCTCCAGGACATCTCGCTCGAGATGATCCTGGATGACGAGGGCCGGAAGGCGCTGTGCTCGGAGTTCCCCAAGATCATGCAGCAGGGTTTCACCTATCTCCCTGGAGGCGTCTGCAAGTCGAGCATGGGGCGGCAGGCATCGTATGAGCAGGCGGTGGCGTGGAAGGTTCTTAGCGACGACGACGCGCCGCACTGCCTCGCCTTCATGCTCGTCAACTGGACCTTCATGTGA。

SEQUENCE LISTING

＜110〉Tianjin Normal University

＜120〉the OsDLS1 gene is in the application aspect adjusting and controlling rice leaf senile and heading stage

<160> 3

<170> PatentIn version 3.5

<210> 1

<211> 7140

<212> DNA

＜213〉OsDLS1 gene

<400> 1

aaataagaaa caacgagact cttgatgtac caagccgagt ctgaagaaga gaaaagagag 60

agagttcaga aaaaggaaac gaccctattc tgatcagaaa aattcaatct ttctctctct 120

ttctctcaga aaaggtcgag agagaaagtg gatgtcttca tgtgaagacc agagagaaga 180

cgagaggcga tcgcagtgga gtggagtgaa gtactagcag tattaaaacc agcagcaagc 240

gagagagagg ccaccaacca cttcctcctc caagaaacca cccccaccac gagcacgagc 300

acgagcaacc atggcggtgg cagagggagc tgcgactgcg agctacgggc catgactgcg 360

gcgagttctt gctgggttgt tgttgtacaa gcagcgacga ggaggagggg agcagaggag 420

gaggaggcgg agggatggca gcggcggcgg tggggggaag gggggagagg ctgtcgtcct 480

cctccccgac ggcggcggcg ccgcaggtgg acgccgggaa gtacgtccgg tacacgcccg 540

agcaggtgga ggcgctggag cgggtgtaca ccgagtgccc caagcccagc tcactccgcc 600

gccagcagct catccgggag tgccccatcc tcagcaacat cgagcccaag cagatcaagg 660

tctggtttca gaatcgcagg tcagtctcat ctccttctct ctcttcgatc tgtttctgct 720

cgggtttgtt cagcagccat ggcgtgtgtt cttcggtttc ttttcttggt tttgtgtaat 780

gtgatgagat tgattgcttt gttgtttgca gttttttttc atgttcttgt gggaatgggg 840

gatcctttgt gaggaagatt tgggttcttg agatgagttt tttttttctg tttgatttgt 900

agatgtctgt aactttttgg gtgttgattt tagggcttgc aatgtgttgg attggtgacg 960

agaaggttca tttctgggat ttgcatccta tatgatgggt tttgtttttt tagttcgtct 1020

tgtgcaataa tgaagcgcaa tctgtttgct tgaaatttat caggatattt tcttttataa 1080

aactttgcgg caatgatttg agttcctgtg cccacactgt tcttcgctgc attattctgc 1140

tgggttgtgt ggtgggtagg gagcgaaatg gcagcatcat tcgttggttt cctcttggat 1200

tttgtggtga tgtgaatagc caaatatcct ggatctctgt acagacttta cagttaattt 1260

tgtttggttt aaattgtaaa tatatgctct ctgattgttt tgttgcttgt ggtgaccaat 1320

gattggttgg agcaaaattg ccttaggtca tttcttgttt cctgtgcttg ccattttaag 1380

ggccatcttt ttaaccctgt gttgtgggct gtggtagtgt tcatgttttc ttgctatagg 1440

cctccataag catcacctcc tggttatcat ctcgctttct tttttcagtg gatcacaagt 1500

agggcaatag ttttgtcact ggtggtgtca ctgtggaaga atgtgaaggg caacattttg 1560

agcaagtttg gtatatacta ttactgtact catgaaggcc aaatataaat caagaatcct 1620

ttttaggtag aggatgctct tgtttagagt tgagttcaca gatgcagaca gaaacatcac 1680

tgtcaattag gcaatgattt tcacatatgg aatactaact gttattgcat agtatatagt 1740

catttgtctc ttttgcaaat gaccttttgc ttgttcctgt tcaatgatcc cccttttaac 1800

cttggggaat atttgctaaa atgcctttgc ctaacttcct ttgtgctcat ttgggctgtg 1860

ccaatcagta tccattaaca agtattggaa tcttgtgtat tagaagacac catcctttag 1920

atagtttatt catgtacttt tgccactgaa attgtgcctt ccttgtaatt ggcagatgcc 1980

gggagaagca gcgcaaggag gcctcccggc tgcaaactgt gaaccggaag ctgaatgcga 2040

tgaacaagct gttgatggag gaaaatgacc ggctgcagaa gcaggtgtcc cgtctcgtct 2100

acgagaacgg ctacatgcgg actcaactgc acaatgtgag ctccctgtgc ttacaggctt 2160

ttgttcttac aattgcttca tattttgatc atcctgtttt cttaatcagt aatgtttgta 2220

tgtactttgt agccttctgc agccaccaca gatacaagct gtgagtctgt ggtgacaagt 2280

ggtcagcacc atcaacagca aaacccagca gttctgcatc cgcaaaggga tgcgaacaac 2340

ccagcagggt atccacaaaa cctggtctat tttcttcttt tgttaggccg tgcttttttg 2400

tatgaaagat tgctgagatg attttatttg cgcagtcttc tcgctattgc tgaggagacc 2460

ttggcagagt tcatgtcgaa ggcgacagga actgctgtcg aatgggtgca aatggttggg 2520

atgaaggtaa tgtattcgtt gtacaagaaa actgtgcagt atgttgattt tctctttcca 2580

ttcatgtttt ttttgttcaa atcaactgaa atgaaaattt gattacagcc tggtccggat 2640

tccattggaa tcatcgctgt ttcgcacaat tgcagtggcg tagcagcacg agcttgcggc 2700

ctggtgagcc ttgagcccac aaaggtatga tttgttttcc ttgatattgt gttacatttc 2760

ccattaactg gcatgctaca gaccctgata agcttcattt taggttgcag agatcctcaa 2820

ggatcgcccc tcttggtacc gtgattgccg atgcgtagat atcatccatg ttatccctac 2880

gggtaacggc ggaaccattg agctaatcta catgcaggta aatataccca atcgtgactc 2940

atgtgttgca tttttctcaa gcatcactag ctactctttg tttttggtaa taaccattgc 3000

ccttttcaat tccttagaca tatgcaccga caactttggc ggcaccacgc gacttttgga 3060

ctctccgcta cacaagtgga cttgaagatg ggagtcttgt ggtattactt tgatctttaa 3120

ttgcatgctg tcagctcatt aaacaaaatg cttcctttca ttttcttgta tggtctattc 3180

gtcatgttat gatcggactt gttctgtttg gatcatacca catcatcatg gttttcaact 3240

gaagtgtagt ttctgttgta ttaaatattg tttgacacca ttcctgttct gtagatctgt 3300

gaaaggtcat tgactcagtc cactggtggc ccatctgggc ctaacactcc gaattttatc 3360

agagccgagg tacttcccag tggttatctg attcgaccat gtgagggagg tggctccatg 3420

atttacattg ttgatcatgt tgatttggat gtaagtaaca atagtgtttt cacattaact 3480

agtgtaatat tgaatagttt tgagatacat atcttattaa tcctgtgtat tttacaggct 3540

tggagtgtgc ctgaggttct tcgaccgctc tatgaatctc caaagatcct tgcacagaag 3600

atgaccattg cagtatgtgc actgttttat tctgttgttg aagagcactt ggtcacctca 3660

tggtgattgt gagtgcataa cttttcttat caccaatgac atggttcatt ctgttcaggc 3720

attgcgacac attaggcaaa ttgcgcatga gtcaagcggt gaaatcccct acggtgctgg 3780

tcgccagcct gctgttttca ggaccttcag tcaaaggctc agcaggtctg tagctatttg 3840

agggttccca tgtattagaa ccaattggca tactgtcata tgacacagtt ccatgcatgt 3900

ctcattgctc acgcatctga actccatact ctaatttgca gaggtttcaa tgatgctgtt 3960

agtggatttc cagatgatgg gtggtctttg ttgagcagtg atggtagtga ggacattaca 4020

atttcagtaa actcgtctcc aaacaagctt gttggatccc atgtcagccc gaaccctctg 4080

ttttctactg ttggaggtgg catcttgtgc gcaaaggcat caatgctact tcaggttgac 4140

ataggctcta tttagttgct cagctaatta tagattgtct ctatctttgt ttcactttga 4200

ccaaaattgt tccaaatcaa agaattaagc tttacaaaat gtcatttttt ttataatgta 4260

gaatgtacct cctgctctac ttgtgcgatt tctgagggag catcgttctg aatgggctga 4320

tcctggtgtt gatgcttatt ctgctgcttc tttgcgagcc agcccatatg cagttccagg 4380

cttaaggacc agcgggttta tgggcagtca ggttatacta cccctcgctc ataccttaga 4440

gcatgaagag gtaatgttgt aattgagtcc ttgtaggctt gtagctcctt ttccacaatg 4500

aatggcaaat attaatgcat caacctggca ttctgaattg cagttcttgg aggttattag 4560

gctcgaagga catggtttca gccatgatga ggtgcttcta tcacgggata tgtaccttct 4620

acaggtattg tggaccatcc gagtcaacta tttattcctt tgctgttggg ctggacaagt 4680

gggcttcggt gggaggcatg ctgctgttct tggccaccgt atgggcctaa ccttctgctc 4740

agggtttctg gtggatcagg tcctcctgcg cacgaattat tttttatttt tgttcataat 4800

aaaagctgag atgtaaaagg tttaagagtc ttagtttagg ttatggtgct tcgttaagtc 4860

tagcagggtg tgctgcagct cgaggacgag ctgctcaaaa aggggggagt aatgtcatgg 4920

gctttgggcc gggcgtccta agtccattgg ataggattag ggtttgttag gattagataa 4980

ggtttgttag gattagatta aggagtcctc catctatata aggagggatc ctatctcagg 5040

tcaattaggc attagatcaa gatttatctt agtgcccatc ggcctgcctt ctcagtgcga 5100

cggagagcgt cgcgccgttt aggttcagga ccgtattcct tgttatttca tctactataa 5160

tccaatctat ccccattcat cccgattctt ctccacataa tccttgctca tgacaattgc 5220

cttaagttgt acctgtttgt tgcattttcc tggttctgag agattgcttt ctatgcagct 5280

ttgcagcggg gttgatgaga atgctactag tgcatctgca cagcttgtct ttgctcctat 5340

tgacgaatct tttgctgatg atgcaccact tttaccctca ggattccgtg tgataccact 5400

agatacaaaa atggtatgac tggtccataa tcttattcca gaggccttgt ctttcccatg 5460

ttggctcacc aattaaccgt tatgcactca aacctgcagg atgggccatc tgcaacacgc 5520

acgctcgacc ttgcatccgc ccttgaggtt ggaccaggtg gagcttcgcg tgcttcagtt 5580

gaagcatctg gcacatgtaa cagatcagtc ctgaccattg ctttccaatt ctcatacgag 5640

aaccaccttc gtgagagcgt ggcggcaatg gctaggagtt atgtcagagc tgtgatggca 5700

tccgtgcaga gggtggctgt ggcaatagct ccttctcgac ttggacccca gatcggaatg 5760

aagcaccctc cagcctctcc tgaggcgctt acacttgcta gctggattgg taggagctat 5820

aggtaagtaa aatacttatt tttagcaaaa gtctatctca ggttcttcat catcagaacc 5880

acctacacat tgccacatct gaagttgtga tcaatgatgt taatgcacta gttatttccg 5940

tggatctttc agggctcaca ccggagcaga tatccgttgg tctgataccg aagatgccga 6000

ttctcccctg gcgctcctgt ggaagcacag cgacgcaata ctgtgctgtt ctctgaaggt 6060

tagtagtgct gcccactaat tttatgctca tttcatcaca tgcatgttct ggaaaccttg 6120

ctaatggtca gtttggttat ctactggcag cctgctccta tgttcacctt cgccaacaat 6180

gccggcctcg acatccttga gacgacgctg gtcaacctcc aggacatctc gctcgagatg 6240

atcctggatg acgagggccg gaaggcgctg tgctcggagt tccccaagat catgcagcag 6300

gtaaaaccac cgtgtccctg cctcgcgaca atcggtgacg catccatcag agtactacca 6360

gaccatgagc aatgttcctg atcaagcctg tttggggtct gtctatcttc tttcagggtt 6420

tcacctatct ccctggaggc gtctgcaagt cgagcatggg gcggcaggca tcgtatgagc 6480

aggcggtggc gtggaaggtt cttagcgacg acgacgcgcc gcactgcctc gccttcatgc 6540

tcgtcaactg gaccttcatg tgatccacca tccatggttt ccagcttcca gctagtgctt 6600

ttgcttttgt gccattgtgg caaactgaac aaagacctga ataccaccac cgtattgtat 6660

taaacatatg cgtccataca tgcatggtgc ttgctttatc ctagctagtg ctagcacctg 6720

tgtgtatggt ctctccatgt ttgaagctta aaattttaga accctggttt ttttttttct 6780

gctgctgtgt ggatgaacaa gtttcaggac ctatgtatct ctatctatct atgctaatgc 6840

tatgaattgt gtgatgtgtg tatgctttga gcaatcggca ttgagttacc ttggttggtt 6900

tatgccgtgc tgaaccattg ttgatgcata tgtagctgaa ggaagtattt gctgaatgtg 6960

attcgtattc ctccaccgtt aaactaaaac aaaagcgttt ctacctatca aaaccggctc 7020

attatgctct cagcttaaac cattatcctt tgctgattgt ggtacatagt tgagggagcc 7080

caaatagact tttctcagtg ctacgtcctg ctcactgatc tcggccacga atagaatcgt 7140

<210> 2

<211> 855

<212> PRT

＜213〉OsDLS1 amino acid

<400> 2

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

1 5 10 15

Ser Pro Thr Ala Ala Ala Pro Gln Val Asp Ala Gly Lys Tyr Val Arg

20 25 30

Tyr Thr Pro Glu Gln Val Glu Ala Leu Glu Arg Val Tyr Thr Glu Cys

35 40 45

Pro Lys Pro Ser Ser Leu Arg Arg Gln Gln Leu Ile Arg Glu Cys Pro

50 55 60

Ile Leu Ser Asn Ile Glu Pro Lys Gln Ile Lys Val Trp Phe Gln Asn

65 70 75 80

Arg Arg Cys Arg Glu Lys Gln Arg Lys Glu Ala Ser Arg Leu Gln Thr

85 90 95

Val Asn Arg Lys Leu Asn Ala Met Asn Lys Leu Leu Met Glu Glu Asn

100 105 110

Asp Arg Leu Gln Lys Gln Val Ser Arg Leu Val Tyr Glu Asn Gly Tyr

115 120 125

Met Arg Thr Gln Leu His Asn Pro Ser Ala Ala Thr Thr Asp Thr Ser

130 135 140

Cys Glu Ser Val Val Thr Ser Gly Gln His His Gln Gln Gln Asn Pro

145 150 155 160

Ala Val Leu His Pro Gln Arg Asp Ala Asn Asn Pro Ala Gly Leu Leu

165 170 175

Ala Ile Ala Glu Glu Thr Leu Ala Glu Phe Met Ser Lys Ala Thr Gly

180 185 190

Thr Ala Val Glu Trp Val Gln Met Val Gly Met Lys Pro Gly Pro Asp

195 200 205

Ser Ile Gly Ile Ile Thr Val Ser His Asn Cys Ser Gly Val Ala Ala

210 215 220

Arg Ala Cys Gly Leu Val Ser Leu Glu Pro Thr Lys Val Ala Glu Ile

225 230 235 240

Leu Lys Asp Arg Pro Ser Trp Tyr Arg Asp Cys Arg Cys Val Asp Ile

245 250 255

Ile His Val Ile Pro Thr Gly Asn Gly Gly Thr Ile Glu Leu Ile Tyr

260 265 270

Met Gln Thr His Ala Pro Thr Thr Leu Ala Ala Pro Arg Asp Phe Trp

275 280 285

Thr Leu Arg Tyr Thr Ser Gly Leu Glu Asp Gly Ser Leu Val Ile Cys

290 295 300

Glu Arg Ser Leu Thr Gln Ser Thr Gly Gly Pro Ser Gly Pro Asn Thr

305 310 315 320

Leu Asn Phe Ile Arg Ala Glu Val Leu Pro Ser Gly Tyr Leu Ile Arg

325 330 335

Pro Cys Glu Gly Gly Gly Ser Met Ile Tyr Ile Val Asp His Val Asp

340 345 350

Leu Asp Ala Trp Ser Val Pro Glu Val Leu Arg Pro Leu Tyr Glu Ser

355 360 365

Pro Lys Ile Leu Ala Gln Lys Met Thr Ile Ala Ala Leu Arg His Ile

370 375 380

Arg Gln Ile Ala His Glu Ser Ser Gly Glu Ile Pro Tyr Gly Ala Gly

385 390 395 400

Arg Gln Pro Ala Val Phe Arg Thr Phe Ser Gln Arg Leu Ser Arg Gly

405 410 415

Phe Asn Asp Ala Val Ser Gly Phe Pro Asp Asp Gly Trp Ser Leu Leu

420 425 430

Ser Ser Asp Gly Ser Glu Asp Ile Thr Ile Ser Val Asn Ser Ser Pro

435 440 445

Asn Lys Leu Val Gly Ser His Val Ser Pro Asn Pro Leu Phe Ser Thr

450 455 460

Val Gly Gly Gly Ile Leu Cys Ala Lys Ala Ser Met Leu Leu Gln Asn

465 470 475 480

Val Pro Pro Ala Leu Leu Val Arg Phe Leu Arg Glu His Arg Ser Glu

485 490 495

Trp Ala Asp Pro Gly Val Asp Ala Tyr Ser Ala Ala Ser Leu Arg Ala

500 505 510

Ser Pro Tyr Ala Val Pro Gly Leu Arg Thr Ser Gly Phe Met Gly Ser

515 520 525

Gln Val Ile Leu Pro Leu Ala His Thr Leu Glu His Glu Glu Phe Leu

530 535 540

Glu Val Ile Arg Leu Glu Gly His Gly Phe Ser His Asp Glu Val Leu

545 550 555 560

Leu Ser Arg Asp Met Tyr Leu Leu Gln Leu Cys Ser Gly Val Asp Glu

565 570 575

Asn Ala Thr Ser Ala Ser Ala Gln Leu Val Phe Ala Pro Ile Asp Glu

580 585 590

Ser Phe Ala Asp Asp Ala Pro Leu Leu Pro Ser Gly Phe Arg Val Ile

595 600 605

Pro Leu Asp Thr Lys Met Asp Gly Pro Ser Ala Thr Arg Thr Leu Asp

610 615 620

Leu Ala Ser Ala Leu Glu Val Gly Pro Gly Gly Ala Ser Arg Ala Ser

625 630 635 640

Val Glu Ala Ser Gly Thr Cys Asn Arg Ser Val Leu Thr Ile Ala Phe

645 650 655

Gln Phe Ser Tyr Glu Asn His Leu Arg Glu Ser Val Ala Ala Met Ala

660 665 670

Arg Ser Tyr Val Arg Ala Val Met Ala Ser Val Gln Arg Val Ala Val

675 680 685

Ala Ile Ala Pro Ser Arg Leu Gly Pro Gln Ile Gly Met Lys His Pro

690 695 700

Pro Ala Ser Pro Glu Ala Leu Thr Leu Ala Ser Trp Ile Gly Arg Ser

705 710 715 720

Tyr Arg Ala His Thr Gly Ala Asp Ile Arg Trp Ser Asp Thr Glu Asp

725 730 735

Ala Asp Ser Pro Leu Ala Leu Leu Trp Lys His Ser Asp Ala Ile Leu

740 745 750

Cys Cys Ser Leu Lys Pro Ala Pro Met Phe Thr Phe Ala Asn Asn Ala

755 760 765

Gly Leu Asp Ile Leu Glu Thr Thr Leu Val Asn Leu Gln Asp Ile Ser

770 775 780

Leu Glu Met Ile Leu Asp Asp Glu Gly Arg Lys Ala Leu Cys Ser Glu

785 790 795 800

Phe Pro Lys Ile Met Gln Gln Gly Phe Thr Tyr Leu Pro Gly Gly Val

805 810 815

Cys Lys Ser Ser Met Gly Arg Gln Ala Ser Tyr Glu Gln Ala Val Ala

820 825 830

Trp Lys Val Leu Ser Asp Asp Asp Ala Pro His Cys Leu Ala Phe Met

835 840 845

Leu Val Asn Trp Thr Phe Met

850 855

<210> 3

<211> 2568

<212> DNA

＜213〉OsDLS1 gene

<400> 3

atggcagcgg cggcggtggg gggaaggggg gagaggctgt cgtcctcctc cccgacggcg 60

gcggcgccgc aggtggacgc cgggaagtac gtccggtaca cgcccgagca ggtggaggcg 120

ctggagcggg tgtacaccga gtgccccaag cccagctcac tccgccgcca gcagctcatc 180

cgggagtgcc ccatcctcag caacatcgag cccaagcaga tcaaggtctg gtttcagaat 240

cgcagatgcc gggagaagca gcgcaaggag gcctcccggc tgcaaactgt gaaccggaag 300

ctgaatgcga tgaacaagct gttgatggag gaaaatgacc ggctgcagaa gcaggtgtcc 360

cgtctcgtct acgagaacgg ctacatgcgg actcaactgc acaatccttc tgcagccacc 420

acagatacaa gctgtgagtc tgtggtgaca agtggtcagc accatcaaca gcaaaaccca 480

gcagttctgc atccgcaaag ggatgcgaac aacccagcag gtcttctcgc tattgctgag 540

gagaccttgg cagagttcat gtcgaaggcg acaggaactg ctgtcgaatg ggtgcaaatg 600

gttgggatga agcctggtcc ggattccatt ggaatcatca ctgtttcgca caattgcagt 660

ggcgtagcag cacgagcttg cggcctggtg agccttgagc ccacaaaggt tgcagagatc 720

ctcaaggatc gcccctcttg gtaccgtgat tgccgatgcg tagatatcat ccatgttatc 780

cctacgggta acggcggaac cattgagcta atctacatgc agacacatgc accgacaact 840

ttggcggcac cacgcgactt ttggactctc cgctacacaa gtggacttga agatgggagt 900

cttgtgatct gtgaaaggtc attgactcag tccactggtg gcccatctgg gcctaacact 960

ctgaatttta tcagagccga ggtacttccc agtggttatc tgattcgacc atgtgaggga 1020

ggtggctcca tgatttacat tgttgatcat gttgatttgg atgcttggag tgtgcctgag 1080

gttcttcgac cgctctatga atctccaaag atccttgcac agaagatgac cattgcagca 1140

ttgcgacaca ttaggcaaat tgcgcatgag tcaagcggtg aaatccccta cggtgctggt 1200

cgccagcccg ctgttttcag gaccttcagt caaaggctca gcagaggttt caatgatgct 1260

gttagtggat ttccagatga tgggtggtct ttgttgagca gtgatggtag tgaggacatt 1320

acaatttcag taaactcgtc tccaaacaag cttgttggat cccatgtcag cccgaaccct 1380

ctgttttcta ctgttggagg tggcatcttg tgcgcaaagg catcaatgct acttcagaat 1440

gtacctcctg ctctacttgt gcgatttctg agggagcatc gttctgaatg ggctgatcct 1500

ggtgttgatg cttattctgc tgcttctttg cgagccagcc catatgcagt tccaggctta 1560

aggaccagcg ggtttatggg cagtcaggtt atactacccc tcgctcatac cttagagcat 1620

gaagagttct tggaggttat taggctcgaa ggacatggtt tcagccatga tgaggtgctt 1680

ctatcacggg atatgtacct tctacagctt tgcagcgggg ttgatgagaa tgctactagt 1740

gcatctgcac agcttgtctt tgctcctatt gacgaatctt ttgctgatga tgcaccactt 1800

ttaccctcag gattccgtgt gataccacta gatacaaaaa tggatgggcc atctgcaaca 1860

cgcacgctcg accttgcatc cgcccttgag gttggaccag gtggagcttc gcgtgcttca 1920

gttgaagcat ctggcacatg taacagatca gtcctgacca ttgctttcca attctcatac 1980

gagaaccacc ttcgtgagag cgtggcggca atggctagga gttatgtcag agctgtgatg 2040

gcatccgtgc agagggtggc tgtggcaata gctccttctc gacttggacc ccagatcgga 2100

atgaagcacc ctccagcctc tcctgaggcg cttacacttg ctagctggat tggtaggagc 2160

tatagggctc acaccggagc agatatccgt tggtctgata ccgaagatgc cgattctccc 2220

ctggcgctcc tgtggaagca cagcgacgca atactgtgct gttctctgaa gcctgctcct 2280

atgttcacct tcgccaacaa tgccggcctc gacatccttg agacgacgct ggtcaacctc 2340

caggacatct cgctcgagat gatcctggat gacgagggcc ggaaggcgct gtgctcggag 2400

ttccccaaga tcatgcagca gggtttcacc tatctccctg gaggcgtctg caagtcgagc 2460

atggggcggc aggcatcgta tgagcaggcg gtggcgtgga aggttcttag cgacgacgac 2520

gcgccgcact gcctcgcctt catgctcgtc aactggacct tcatgtga 2568

Claims (1)

1.HD-Zip III family member OsDLS1The application of gene in delaying the rice leaf aging, described OsDLS1The aminoacid sequence of genes encoding is shown in SEQ ID No.2.

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