CN110029125A

CN110029125A - Paddy gene ORYsa;The genetic engineering application of SQD1

Info

Publication number: CN110029125A
Application number: CN201910213757.1A
Authority: CN
Inventors: 孙淑斌; 孙雅菲; 王小文
Original assignee: Nanjing Agricultural University
Current assignee: Nanjing Agricultural University
Priority date: 2019-03-20
Filing date: 2019-03-20
Publication date: 2019-07-19
Anticipated expiration: 2039-03-20
Also published as: CN110029125B

Abstract

The invention discloses Deficiency Xn Rice Plants phosphorus responsive genes ORYsa；The genetic engineering application of SQD1.Including the gene in terms of improving Phosphorus in Soil absorption and use efficiency and improve rice yield and influence seed viability application.Deficiency Xn Rice Plants phosphorus responsive genes ORYsa；The cDNA sequence SEQ ID NO.1 of the SQD1 and its amino acid sequence SEQ ID NO.2 of coding.Gene of the present invention is the report for the first time in monocotyledon, is played an important role in terms of improving Phosphorus in Soil absorption and use efficiency and yield.

Description

Paddy gene ORYsa；The genetic engineering application of SQD1

Technical field

The present invention relates to field of plant genetic project technology, are related to paddy gene ORYsa；The genetic engineering application of SQD1.

Background technique

Rice has extensive plantation in the whole world, is the most important cereal crops in Asia, and the even more main grain in China is made One of object, China's Monitoring of Paddy Rice Plant Area account for the 1/3 of global cereal crops cultivated area, and rice yield accounts for national total grain output 50%, it is the most important crop (Hu Peisong etc., 2002) for guaranteeing China's grain security.

Phosphorus is the nutrition member of very easy shortage in one of macronutrient needed for plant growth and much soil One of element.Although the total phosphorus content in soil is often very high, P use efficiency is only the pass of limiting plant growth and productivity Key factor.It is this it is contradictory occur be because the soil liquid in titanium pigment mean concentration be about 1 μM, rarely exceed 10 μM (Bieleski, 1973) much can not meet the growth needs of plant, become a big limitation sex factor of plant growth. Therefore can plant efficiently have vital influence to plant growth using titanium pigment a small amount of in soil.

For a long time, continuous exploitation and utilization of the people to this non-renewable resources of phosphorus, so that the utilizability of phosphorus will Peak-peak is reached in the year two thousand thirty.This will cause in partial development country's agriculturally bigger economic input.On the other hand, While alleviation crops lack phosphorus, the not scientifical use of phosphate fertilizer leads to the decline of soil quality, although compared with the past, agricultural The absolute magnitude of phosphorus no longer lacks in soil, but very low using efficiency.The New Crop Varieties initiative of High-Efficiency Utilization of Phosphorus can subtract Few phosphate fertilizer, improves P use efficiency, and all having to agricultural production and ecological environment has very great meaning (Wu et al.,2013；Lopez-Arredondo et al.,2014).

Thio isorhamnose and thio isorhamnose diglyceride synthase gene in research before, in arabidopsis SQD1 is to be participated in the regulation process of dynamic equilibrium between phosphatide and thioester under the conditions of scarce phosphorus, but not by phosphorus starvation induced gene Disclosing the gene has the function of promoting arabidopsis growth.

Summary of the invention

The purpose of the present invention is to provide the genetic engineering applications of Deficiency Xn Rice Plants phosphorus responsive genes OsSQD1.

The purpose of the present invention can be achieved through the following technical solutions:

The present invention provides the genetic engineering application of Deficiency Xn Rice Plants phosphorus responsive genes OsSQD1, gene order accession number is Os05g0387200.For the function of studying OsSQD1, it is thus necessary to determine that and clone and obtain the accurate sequence of OsSQD1.By according to number Sequence is provided according to library and carries out RT-PCR, and clone has obtained the complete coding region sequence of OsSQD1, encodes region nucleotide sequence such as SEQ Shown in ID NO.1, the amino acid sequence of coding is as shown in SEQ ID NO.2.OsSQD1 gene contains 2 exons altogether, in 1 Containing son, code area overall length 1440bp encodes 479 amino acid.According to the sequence being cloned into, we construct the super table of OsSQD1 Up to rotaring carrier, and rice transformation, raise its expression quantity.

The homozygote strain of acquisition is subjected to solution culture, measures seedling stage different parts total phosphorus concentration, can extract phosphorus concentration Etc. physical signs.

The homozygote strain of acquisition is subjected to potted plant experiment, measurement is sampled in maturity period Divisional, and it is dense to measure each position phosphorus Spend difference.Sampling sites include: down a leaf, two leaves, leaf sheath, stem, fringe handle and seed.

Said gene ORYsa；The genetic engineering application of SQD1 is intended to improve rice P acquisition and utilize, Yi Jiti The tolerance that high rice coerces phosphorus element in environment.

Beneficial effect

All there are many different from the response mechanism of dicotyledonous model plant arabidopsis phosphorus as unifacial leaf model plant for rice. Gene ORYsa of the present invention；The function of SQD1 is the report for the first time in monocotyledon.We pass through in multiple genome databases In retrieval compare, it is determined that the homologous target gene of SQD1 in rice, and being obtained in rice by the means of PCR clone The code area complete sequence of OsSQD1 gene.We identify the expression pattern of OsSQD1 gene by molecular biology method With subcellular localization feature.The carrier of SQD1-Ox is constructed, the expression of excessive SQD1 passes through a series of identification of Physiological Experiments It was found that: while OsSQD1 overexpression increases the width of seed, improve rice yield；At the same time, soil is being improved It plays an important role in terms of middle P acquisition utilization efficiency.Titanium pigment contains under the conditions of also improving vegetative growth of rice plants phase scarce phosphorus The content of tatal phosphorus of amount and reproductive stage stem, fringe and seed, and significantly improve to experience outside rice and lack phosphorus signal Susceptibility improves the efficiency that phosphorus element is reallocated in rice, suitable to cultivate to improve plant tolerant to low-phosphorus ability, nutrientuse efficiency New rice variety for phosphorus lean soil provides guarantee.

Detailed description of the invention

Fig. 1: paddy gene ORYsa；Expression pattern of the OsSQD1 at different tissues position.

Fig. 2: paddy gene ORYsa；Expression pattern of the OsSQD1 at different scarce phosphorus time points.

Fig. 3: the phenotype of wild type and OsSQD1 overexpression material full ripe stage.

Fig. 4: in potted plant experiment, the seed morphology and mass of 1000 kernel of wild type and OsSQD1 overexpression material.

Fig. 5: the phenotype and biomass of wild type and overexpression material under the conditions of normal phosphorus supply and scarce phosphorus.

Fig. 6: wild type and OsSQD1-Ox material are in the full phosphorus of vegetative growth phase different parts under different phosphate treatment conditions With titanium pigment content.

Fig. 7: the full phosphorus under the conditions of maturity period normal phosphorus supply and scarce phosphorus in wild type and OsSQD1-Ox strain different parts contains Amount.

Fig. 8: wild type and OsSQD1-Ox material overground part and root Pht1 family member under the conditions of normal phosphorus supply and scarce phosphorus Expression.

Fig. 9: pTCK303 plasmid map.

Specific embodiment

The acquisition of embodiment 1, gene order

AtSQD1 is one and is located on Article 5 chromosome that coding synthesizes thio isorhamnose diglyceride synthase SQD1 Gene.SQD1 enzyme is in the nature short-chain dehydrogenase superfamily (Short-Chain Dehydrogenase superfamily； SDR uridine 5'-diphosphate thio isorhamnose synthase family (the UDP-sulfoquinovose synthase family in)； SDR52E a member in) has NAD⁺Binding site, can in conjunction with ferredoxin glutamine synthase, and receive from Inferior sulfate radical (the SO that the external world is transmitted to₃ ^2-).Homology search is carried out in NCBI according to arabidopsis SQD1 gene order, in NCBI Homologous gene OsSQD1 in rice is obtained in database, gene number is (same to RAB_DB) Os05g0387200.By Blast and GenomeScan analysis shows: OsSQD1 is located on rice Article 5 chromosome, specific location Chr5: 18738602..18741791.Rice SQD1 gene contains 2 exons and 1 introne, and the protein-coding region of gene is 1440bp encodes 479 amino acid.

Embodiment 2, ORYsa；The expression pattern of SQD1 gene is identified

2.1, the extraction of total serum IgE and transcription synthesis the first chain of cDNA

It selects rice varieties " OryzasativaLcv.Nipponbare ", after rice seedling long to 10 days, 1 week post-sampling of complete nutrition liquid culture is ( One leaf, two leaf, four leaf, two leaf leaf sheath, four leaf leaf sheath, rhizome engaging portion, the root system tip of a root, root elongation area and maturation Area).Rice seedling age to transplanting in 30 days carries out potting processing into basin Portland, and seedling age 60 days whens are sampled (boot leaf, three leaf, flag Leaf leaf sheath, three leaf leaf sheath, bar, stem knot and fringe handle).TriZol reagent extracted total RNA is respectively adopted, uses agarose gel electrophoresis It identifies total serum IgE quality, rna content is then measured on spectrophotometer.Using the total serum IgE of acquisition as template, obtained by reverse transcription The first chain of rice cDNA is obtained, is used for subsequent experiment.The synthesis step of the first chain of cDNA: with the water-treated PCR pipe of DEPC (300 μ l), is added 5 μ g, oligodT 1 μ L (25ng/ μ L), dNTP (10mmoL/L) 2 μ L of total serum IgE, 65 DEG C of water-bath 5min, fast Speed is placed in cooled on ice, adds 5 × reaction buffer 4 μ L, M-MLV reverse transcriptase (200U/ μ L) 1 μ L, RNase inhibitor, 0.5 μ L, DEPC water to total volume is 20 μ L (operating on ice above).Slightly centrifugation is placed on 42 DEG C of water-baths 1h, 70 DEG C of water-bath 10min, It is subsequently placed in cooling rapidly on ice.Products therefrom is cDNAs, is placed in -20 DEG C of preservations.

2.2 ORYsa；The tissue-specific expression pattern of SQD1 gene is identified

" OryzasativaLcv.Nipponbare " cDNA obtained using step 2.1 is template, according to the sequence of the 3 ' parts UTR of rice Os SQD1 gene Column, designing following OsSQD1 gene specific primer P1, P2 amplification length is that 117bp fragment length identifies ORYsa；OsSQD1 gene Expression pattern.

P1 CTACTACAACGCCAAGCA(SEQ ID NO.3)

P2 ACCAGCCTCCTCTTAGAC(SEQ ID NO.4)

PCR specific steps are as follows: the cDNA obtained using step 1 is template, using ABI StepOnePlus (purchased from ABI biology Company) and SYBR PremixExTaq Kit (being purchased from TaKaRa company) progress gene quantification PCR analysis.According to TaKaRa reagent Illustrate the reaction system using 20 μ l: 10 μ l of SYBR PremixExTaq, each 2 μ l of 0.4 μ l, cDNA template of upstream and downstream primer, 0.4 6.8 μ l of μ l, ddH2O of ROX Reference Dye (50x).Reaction condition are as follows: 40 circulations, 95 DEG C of 1min, 95 DEG C of 5s, 60 DEG C of 30s, 95 DEG C of 15s, 60 DEG C of 1min, 95 DEG C of 5s.Quantitative result analyzes and identifies OsSQD1 gene according to the calculating of 2- Δ Ct method Spatial and temporal expression profile, the result is shown in Figure 1.It entrusts Jin Sirui biotech company in Nanjing to be sequenced and determines that sequence is ORYsa；OsSQD1 Segment.

Fig. 1 can be seen that in the vegetative growth of rice plants stage, and the relative expression quantity of the 3rd week breeding time and the 6th week OsSQD1 are equal Be overground part be higher than root, wherein the 3rd week different leaves (first and second and newest expansion leaf) relative expression quantity without obviously it is poor Different, rhizome junction and root expression quantity are significantly lower than blade (Figure 1A)；Expression quantity of the OsSQD1 equally in blade in 6th week Highest, wherein old leaf (the first leaf and third leaf) expression quantity is apparently higher than most young leaves (Figure 1A).In generative growth phase, breeding time OsSQD1 is mainly expressed in rhizome junction, leaf sheath, blade and immature flower spike within 12nd week, and the expression in root Measure relatively low (Figure 1B)；Breeding time the 14th week, expression quantity highest of the OsSQD1 in blade, bennet and floral axis, in addition to this, leaf Sheath, stipes take second place, and the expression quantity of root system and rhizome junction is minimum (Figure 1B).

2.3 ORYsa；The response of SQD1 gene lacks the expression pattern identification of phosphorus

In order to understand the sensitivity that OsSQD1 expression responds scarce phosphorus, different scarce phosphorus time point (1 day, 3 are set It, 5 days, 7 days and phosphorus 1 day is covered after 7 days), detect its relative expression quantity.It was found that OsSQD1 is either in root or ground Portion, expression quantity variation when scarce phosphorus is handled 1 day are not obvious, and the relative expression quantity of OsSQD1 occurs aobvious since third day The up-regulation of work, and upper modulation with processing the time passage obviously increase, when handling 7 days overground part OsSQD1 expression quantity raise About 5.5 times, root about raises 6 times (Fig. 2).It lacks after phosphorus covers phosphorus element 1 day after 7 days, OsSQD1 expression quantity portion and root on the ground The expression quantity in portion drops to the level (Fig. 2) when normal phosphorus supply again.The result shows OsSQD1 to the level of supply of phosphorus element compared with For sensitivity, and respond scarce phosphorus signal in short term.

Embodiment 3, ORYsa；The acquisition and Function Identification of SQD1 overexpression material

The building of 3.1 OsSQD1 overexpression vectors

In order to construct OsSQD1 overexpression material, we select this laboratory common carrier pTCK303 as expression vector (Fig. 9).According to the cDNA sequence for the OsSQD1 that the website NCBI is announced, with DNAMAN being limited property endonuclease analysis, and Do not have in selection cDNA and KpnI and SpeI that carrier polyclone enzyme enzyme site has, it can be by target fragment directly and pTCK303 Carrier carries out digestion and enzyme even operates.Primer is respectively as shown in table (table 1).Using OryzasativaLcv.Nipponbare leaf cDNA as template, high-fidelity is used PCR Mix carries out PCR amplification and obtains the full length cDNA sequence segment of OsSQD1.PCR product is purified and recycled, clone is connected into and carries Body pEASY-Blunt.After being sequenced and determining sequence and OryzasativaLcv.Nipponbare OsSQD1 sequence indifference, with KpnI and SpeI endonuclease digestion The cloning vector of recombination, the endonuclease bamhi of acquisition is purified with after recycling, is connected to expression vector, and be transformed into Escherichia coli DH5 α competent cell.Positive monoclonal plasmid is extracted, after digestion verification and sequencing ensure correctly, recombinant plasmid is turned through electricity Change and imports in Agrobacterium EHA105.

1 OsSQD1 over-express vector of table constructs the primer

The acquisition and Function Identification of 3.2 overexpression transgenic plants

The Agrobacterium for turning to have expression vector that step 1 is obtained, further conversion to rice is (using Agrobacterium tumefaciens mediated The expression vector of building is transferred to rice OryzasativaLcv.Nipponbare kind by method).Inducing paddy rice mature embryo callus will grow to a certain size water Rice callus is chosen, and is put into agrobacterium suspension and infects 5min (callus amount there was not 50ml centrifuge tube taper position, did not stopped Shake)；Callus is taken out, is placed on sterile filter paper and drains 30-40 minutes；Callus, which is placed in, to be co-cultured on base, 28 DEG C dark culture 2.5 days.Then callus is transferred to the selection culture of carbenicillin containing 250mg/L (Car) and 50mg/L hygromycin It is screened on base.The kanamycin-resistant callus tissue of picking color cadmium yellow moves into the culture dish equipped with differential medium or differentiation tank, is put into Constant temperature incubation room seedling differentiation.Place into one to two week of strong sprout in root media, i.e. acquisition transgenic plant.

3.3 OsSQD1 overexpression affects rice yield

Pass through the observation (Fig. 3) to overexpression strain (Ox1, Ox2 and Ox3) breeding time phenotypic evaluation and fringe form, discovery In rice full ripe stage, overexpression and the phenotype of wild type have differences (Fig. 3 A).The plant height of Ox1, Ox2 and Ox3 are significantly lower than Wild type (Fig. 3 B), tiller number are then obviously (Fig. 3 C) more than wild type.To fringe form it has been observed that OsSQD1 is overexpressed Phenotype no significant difference (Fig. 3 D) between material and wild type tassel, and also no significant difference (Fig. 3 E) of setting percentage between the two.

3.4 OsSQD1 overexpression affects rice paddy seed development

We identify wild type and overexpress the seed phenotypes (Fig. 4 A) of material, overexpress the seed of Ox1, Ox2 and Ox3 Width is obviously wider than wild type (Fig. 4 B), and mass of 1000 kernel is also apparently higher than wild type (Fig. 4 C).The result shows OsSQD1 Overexpression can increase seed width, and then influence seed weight, it is known that OsSQD1 is positive regulation on influencing seed development.

Influence of 3.5 OsSQD1 to the lower growth and development of rice difference phosphorus element processing

In order to probe into function of OsSQD1 vegetative growth phase under the conditions of normal phosphorus supply (+P) and scarce phosphorus (- P), we are right Progress+P (200 μM) and-P (10 μM) water planting are handled 2 weeks long to 10 days seedling of wild type and overexpression strain respectively.As a result it shows Show, the root system for overexpressing strain becomes more flourishing (Fig. 5 A and B) compared with wild type.The plant height and wild type difference of overexpression It is not obvious (Fig. 5 C), root long has notable difference, wherein overexpression rises about 13.8% under+P-condition, overexpresses about under-P-condition Rise 17.5% (Fig. 5 D).Biomass is overexpressed compared with wild type, on the ground portion and root have obvious rising (Fig. 5 E and F)。

3.6 OsSQD1 overexpression improves titanium pigment content under the conditions of Deficiency Xn Rice Plants phosphorus

In order to verify influence of the OsSQD1 to phosphorus content in vegetative growth of rice plants stage body, the phosphorus content in material is overexpressed Also slightly have difference (Fig. 6) with wild type under different disposal.Under+P-condition, titanium pigment and content of tatal phosphorus in overexpression are all Do not change, but under-P-condition, compared with wild type, the titanium pigment content in overexpression obviously rises, and content of tatal phosphorus Have no significant change (Fig. 6).The result shows the overexpressions of OsSQD1 under-P-condition may promote internal organic phosphorus transformation For the process of Phos, just cause under the premise of total phosphorus does not change in vivo, obvious rise occurs in titanium pigment.In summary As a result, thus it is speculated that OsSQD1 mainly has an impact rice overground part and root phosphorus concentration under-P-condition, and may promote body The interior organic phosphorus transformation to Phos.

3.7 OsSQD1 influence rice maturity phosphorus element and reallocate

Pass through the wild type and OsSQD1 overexpression material (Ox1, Ox2 and Ox3) under right+P and-P-condition in the maturity period The measurement of phosphorus content in young leaves (the 6th leaf), old leaf (the second leaf), stem, boot leaf leaf sheath, fringe and seed, under discovery+P processing Phosphorus concentration is without difference in young leaves, old leaf and boot leaf leaf sheath, and overexpresses the phosphorus concentration in material stem and be above wild type (figure 7)；Phosphorus concentration decline in material old leaf is overexpressed under P-condition, the phosphorus concentration in young leaves and boot leaf leaf sheath does not change, and stem Phosphorus concentration in stalk, fringe and seed is apparently higher than wild type (Fig. 7), these results illustrate that OsSQD1 overexpression promotes breeder The accumulation of official's phosphorus element, and inhibit phosphorus element accumulation in old leaf under-P-condition.

3.8 OsSQD1 overexpressions improve response of the rice to environment Low phosphorus stress

In order to probe into influence of the overexpression of OsSQD1 to rice phosphate transport protein Pht1 family member, by glimmering The means of Fluorescent Quantitative PCR, we have detected Pht1 family member in wild type and OsSQD1-Ox material (OsSQD1-Ox1, Ox2 And Ox3) relative expression quantity (Fig. 8) in normal phosphorus supply (+P) and scarce phosphorus (- P) overground part and root.As a result, it has been found that+P is handled Overground part in PT gene relative expression quantity compared with wild type no significant difference, overexpress material (OsSQD1-Ox1, Ox2 The obviously up-regulation (Fig. 8) compared with wild type with the relative expression quantity of the OsPT1 and OsPT4 of Ox3)-P processing overground part；With it is wild Type is compared, and the relative expression quantity of the overexpression material root OsPT1 and OsPT2 of+P processing obviously rises, the super table under-P processing There is different degrees of up-regulation (Fig. 8) up to root OsPT2,4 and 6.It can be seen that the overexpression of OsSQD1 is to Pht1 family The expression of gene is whole to be presented the trend promoted, and promotes more significant under the acting on of Pht1 family gene expression-P-condition. This result also partial interpretation why-P-condition under overexpression in phosphorus content rise.

In conclusion the ORYsa that the present inventor provides；The engineer application of OsSQD1 is to report for the first time in rice.ORYsa； The overexpression gene of OsSQD1 imports plant, and plant phosphorous use efficiency is increased while improving rice yield and absorbs speed Rate provides guarantee to cultivate the new rice variety that high yield, high P acquisition and its internal phosphorus element are efficiently reallocated.In order to simple For the sake of, ORYsa in the present invention；OsSQD1 is labeled as OsSQD1 sometimes.

Sequence table

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aatctagtgc tcaaatcatg tactgcgaca agaccgaaca ggccatttgt cgctcgtgct 180

agcgctgctg tacaaggaca gacacaaaca ccccttactg gaagtcagca agcatctggg 240

cactcatcct ctaaacccaa aaaggtcatg gttatcggcg gagatggcta ctgcggctgg 300

gcaaccgcac ttcatctctc caataaagga tacgaggttg ccattgttga caatcttgtg 360

cggcgccttt ttgatcacca acttggcctg gattccctca cacccatagc ctccatccag 420

aatcgcatcc gtcggtggaa ggctctaacc ggaaagacga ttcagctcta tgttggtgat 480

atatgtgact ttgatttcct ttcagaagcc ttcaagtctt tcgagccgga ttctgctgtc 540

cactttggtg agcagagatc agcgccatat tctatgattg atcgttcccg tgcggtattc 600

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aagcaagcga gctccttcta ccatctaagc aaagtgcatg actcgcacaa tatagcattt 840

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agaacagatg aaactgcaat gcatgaagaa ctatccaaca ggtttgatta tgatggcgtc 960

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gagctcgcaa tcgccaaccc agccaaacca ggtgagttca gggtcttcaa ccagttcacc 1140

gagcagttct cggtcaacga gctggcaaag ctggttacgg ccgccggtgc aaagcttggg 1200

ctggaggtgc agaccaagtc ggtgcccaac ccgcgggtgg aagcggagga acactactac 1260

aacgccaagc acaccaagct catggagctc ggcctggagc cccacctgct gtcggactcg 1320

ctcctcgact cgctgctcaa cttcgccgtc cagtacaagg acagggtcga cacggcgcag 1380

atcatgccca gcgtgtcgtg gaagaagatg ggggcgaagc cgaagacggt gtccgtctaa 1440

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Ala Val Lys Thr Tyr Ser Lys Ser Pro Gly Tyr Cys Cys Asn Val Thr

20 25 30

Gln Phe Gln Ser Ser Lys Cys Ser Asn Leu Val Leu Lys Ser Cys Thr

35 40 45

Ala Thr Arg Pro Asn Arg Pro Phe Val Ala Arg Ala Ser Ala Ala Val

50 55 60

Gln Gly Gln Thr Gln Thr Pro Leu Thr Gly Ser Gln Gln Ala Ser Gly

65 70 75 80

His Ser Ser Ser Lys Pro Lys Lys Val Met Val Ile Gly Gly Asp Gly

85 90 95

Tyr Cys Gly Trp Ala Thr Ala Leu His Leu Ser Asn Lys Gly Tyr Glu

100 105 110

Val Ala Ile Val Asp Asn Leu Val Arg Arg Leu Phe Asp His Gln Leu

115 120 125

Gly Leu Asp Ser Leu Thr Pro Ile Ala Ser Ile Gln Asn Arg Ile Arg

130 135 140

Arg Trp Lys Ala Leu Thr Gly Lys Thr Ile Gln Leu Tyr Val Gly Asp

145 150 155 160

Ile Cys Asp Phe Asp Phe Leu Ser Glu Ala Phe Lys Ser Phe Glu Pro

165 170 175

Asp Ser Ala Val His Phe Gly Glu Gln Arg Ser Ala Pro Tyr Ser Met

180 185 190

Ile Asp Arg Ser Arg Ala Val Phe Thr Gln His Asn Asn Val Ile Gly

195 200 205

Thr Leu Asn Val Leu Phe Ala Ile Lys Glu Phe Ser Glu Glu Cys His

210 215 220

Leu Val Lys Leu Gly Thr Met Gly Glu Tyr Gly Thr Pro Asn Ile Asp

225 230 235 240

Ile Glu Glu Gly Phe Ile Thr Ile Thr His Asn Gly Arg Thr Asp Thr

245 250 255

Leu Pro Tyr Pro Lys Gln Ala Ser Ser Phe Tyr His Leu Ser Lys Val

260 265 270

His Asp Ser His Asn Ile Ala Phe Thr Cys Lys Ala Trp Gly Ile Arg

275 280 285

Ala Thr Asp Leu Asn Gln Gly Val Val Tyr Gly Val Arg Thr Asp Glu

290 295 300

Thr Ala Met His Glu Glu Leu Ser Asn Arg Phe Asp Tyr Asp Gly Val

305 310 315 320

Phe Gly Thr Ala Leu Asn Arg Phe Cys Val Gln Ala Ala Val Gly His

325 330 335

Pro Leu Thr Val Tyr Gly Lys Gly Gly Gln Thr Arg Gly Tyr Leu Asp

340 345 350

Ile Arg Asp Thr Val Gln Cys Val Glu Leu Ala Ile Ala Asn Pro Ala

355 360 365

Lys Pro Gly Glu Phe Arg Val Phe Asn Gln Phe Thr Glu Gln Phe Ser

370 375 380

Val Asn Glu Leu Ala Lys Leu Val Thr Ala Ala Gly Ala Lys Leu Gly

385 390 395 400

Leu Glu Val Gln Thr Lys Ser Val Pro Asn Pro Arg Val Glu Ala Glu

405 410 415

Glu His Tyr Tyr Asn Ala Lys His Thr Lys Leu Met Glu Leu Gly Leu

420 425 430

Glu Pro His Leu Leu Ser Asp Ser Leu Leu Asp Ser Leu Leu Asn Phe

435 440 445

Ala Val Gln Tyr Lys Asp Arg Val Asp Thr Ala Gln Ile Met Pro Ser

450 455 460

Val Ser Trp Lys Lys Met Gly Ala Lys Pro Lys Thr Val Ser Val

465 470 475

<210> 3

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ctactacaac gccaagca 18

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accagcctcc tcttagac 18

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tcggtaccat gaaaatggca catat 25

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tcactagtta gacggacacc gtctt 25

Claims

1. Deficiency Xn Rice Plants phosphorus responsive genes ORYsa；Application of the SQD1 in raising rice phosphorus element is absorbed and utilized, the Deficiency Xn Rice Plants phosphorus Responsive genes ORYsa；SQD1 gene is Os05g0387200 in the accession number of Genbank.

2. Deficiency Xn Rice Plants phosphorus responsive genes ORYsa；SQD1 is improving rice yield and is influencing to apply in seed development, the rice Lack phosphorus responsive genes ORYsa；Accession number Os05g0387200 of the SQD1 gene in Genbank.

3. Deficiency Xn Rice Plants phosphorus responsive genes ORYsa；The rice that SQD1 efficiently reallocates in the high P acquisition of exploitation and its internal phosphorus element The application of new varieties.

4. Deficiency Xn Rice Plants phosphorus responsive genes ORYsa；SQD1 is in the engineer application for improving phosphate transport protein function, the rice Lack phosphorus responsive genes ORYsa；Accession number Os05g0387200 of the SQD1 gene in Genbank.