CN101570791B - Special primer for identifying which variety of Glu-A3 protein subunit is carried in wheat as well as application thereof - Google Patents
Special primer for identifying which variety of Glu-A3 protein subunit is carried in wheat as well as application thereof Download PDFInfo
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- CN101570791B CN101570791B CN2009100861334A CN200910086133A CN101570791B CN 101570791 B CN101570791 B CN 101570791B CN 2009100861334 A CN2009100861334 A CN 2009100861334A CN 200910086133 A CN200910086133 A CN 200910086133A CN 101570791 B CN101570791 B CN 101570791B
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Abstract
The invention discloses a special primer for identifying which type of Glu-A3 protein subunit is carried in wheat as well as an application thereof. The provided special primer comprises at least one pair of the following seven pairs of primers: the primer pair constituted by nucleotides shown in sequence 18 and 20; the primer pair constituted by the nucleotides shown in sequence 19 and 21; the primer pair constituted by the nucleotides shown in sequence 18 and 22; the primer pair constituted by the nucleotides shown in sequence 19 and 23; the primer pair constituted by the nucleotides shown in sequence 18 and 24; the primer pair constituted by the nucleotides shown in sequence 18 and 25; and the primer pair constituted by the nucleotides shown in sequence 18 and 26. The utilization of thespecial primer, a reagent box or a method of the invention can screen the variety of the wheat at the early stage of development, thereby optimizing the hybrid combination, accelerating the breedingspeed and reducing the breeding cost. The method has simple operation, stability, reliability and low cost, thereby being applicable to popularization and having important significance for researchingthe related characters of the wheat quality and improving the wheat quality. The invention will play an important role in wheat quality breeding.
Description
Technical field
The present invention relates to differentiate that wheat is the primer special and the application thereof of carrying which kind of Glu-A3 protein protomer kind.
Background technology
The wheat storage protein mainly comprises glutenin and prolamine.The difference of the two is that the former can form multichained construction through intermolecular disulfide bond, and the latter then exists with monomer subunit form.Glutenin accounts for 35～45% of seed storage protein, is one of staple of gluten.The peptide interchain disulfide linkage and the intramolecular disulfide linkage of glutenin can mutually combine, and mostly its protein is the polare Aminosaeren composition in addition, very easily forms macromole aggregate (Polymer), makes dough have certain elasticity.Good elasticity and ductility are the bases that makes high quality bread, so selection contains good HMW-GS and LMW-GS is one of major objective of marker assisted selection in the Wheat Quality Improvement.According to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Sodium-dodecyl-sulphatepolyacrylamide gel electrophoresis; SDS-PAGE) mobility; Glutenin can be divided into hmw glutenin subunit (High-molecular-weight glutenin subunits; HMW-GS) and low molecular weight glutenin subunit (Low-molecular-weight glutenin subunits, LMW-GS).HMW-GS is respectively by the genes encoding that is positioned at the first homology cohort karyomit(e) long-armed Glu-A1, Glu-B1 and Glu-D1 site (general designation Glu-1); LMW-GS is then respectively by the Glu-A3 that is positioned at galianconism, Glu-B3 and Glu-D3 site genes encoding (Payne etc., 1987 of (being referred to as Glu-3); Singh and Shepherd, 1988).Effective choice need be based upon on the basis to the evaluation of different subunits and evaluation, to satisfy different breeding objectives.
Because the LMW-GS number is more, molecular weight is little and overlapped with a large amount of prolamines on electrophoretogram, traditional unidirectional SDS-PAGE is difficult to it is distinguished.After Singh and Shepherd (1988) had developed two step SDS-PAGE methods, this problem was just solved largely.In ensuing research; RPLC (Reversed-phase high performance liqnid chromatography; RP-HPLC) and the capillary vessel electrophoretic technique (Capillary electrophoresis CE) also is used to differentiate different low molecular weight glutenin subunits (Bean and Lookhart, 2000); But since these technology comparatively complicacy or application charges are higher, all not by large-scale application.Therefore to LMW-GS and and Q factor between the research that concerns fall behind more than HMW-GS.
Have confidential relation between LMW-GS and the wheat quality, the influence of different subunits also is not quite similar.Payne etc. (1984) connect the quality trait of low molecular weight glutenin subunit and tetraploid for the first time.Branlard and Felix (1994) think that HMW-GS can explain 18%～55% of gluten strength and visco-elasticity variation, and LMW-GS can explain 20% of its variation.Thereby, when the concerning of research glutenin and wheat quality, should take all factors into consideration the effect of HMW-GS and LMW-GS.The research of Cornish etc. (1993) shows, subunit combinations is that the ductility of kind of 2*, 7+8,2+12, Glu-A3b, Glu-B3b and Glu-D3b is best.Also have some researchs to think; For different Glu-3 subunits, the kind tool that is combined as Glu-A3b, Glu-B3b and Glu-D3b is best ductility, and subunit combinations is that the extensibility of kind of Glu-A3b, Glu-B3b and Glu-D3c is also better; Subunit combinations is Glu-A3b, Glu-B3b and Glu-D3b; Glu-A3b, Glu-B3b and Glu-D3c, and the kind of Glu-A3c, Glu-B3b and Glu-D3c, its bread quality is best.More detailed research has been done to Chinese wheat breed Glu-1 and Glu-3 site allelic variation in (2005) such as He etc. (2005) and Liu; The relation of clear and definite Glu-A3 and Glu-B3 and bread and noodle quality; Think the effect of d and b allelotrope dough ductility in Glu-B3 site greater than other allelotrope, Glu-A3d and Glu-B3d are a bit larger tham other allelotrope to the contribution of Chinese extra dry white wine noodle quality.
Molecule marker is a kind of genetic marker that polymorphum designed of utilizing with objective trait close linkage or relevant gene.Molecule marker does not receive genetic effect and Effect of Environmental, and is lower to technical requirements, thereby the result is reliable, and can select in generation morning, thereby shortens breeding cycle greatly.Lack easy, effective authentication method, limited analysis and utilization LMW-GS.Thereby the relevant mark of exploitation LMW-GS is for carrying out the LMW-GS functional study, and molecular marker assisted selection (Marker-assistant selection MAS) has significance.
Summary of the invention
The purpose of this invention is to provide and differentiate that wheat is the primer special and the application thereof of carrying which kind of Glu-A3 protein protomer kind.
Discriminating wheat provided by the invention is the primer special that carries the wheat breed of which kind of Glu-A3 protein protomer, comprises in following seven pairs of primers at least one pair of:
(1) primer formed of Nucleotide shown in the sequence 20 of Nucleotide and sequence table shown in the sequence 18 of sequence table is to (gluA3a primer to);
(2) primer formed of Nucleotide shown in the sequence 21 of Nucleotide and sequence table shown in the sequence 19 of sequence table is to (gluA3b primer to);
(3) primer formed of Nucleotide shown in the sequence 22 of Nucleotide and sequence table shown in the sequence 18 of sequence table is to (gluA3ac primer to);
(4) primer formed of Nucleotide shown in the sequence 23 of Nucleotide and sequence table shown in the sequence 19 of sequence table is to (gluA3d primer to);
(5) primer formed of Nucleotide shown in the sequence 24 of Nucleotide and sequence table shown in the sequence 18 of sequence table is to (gluA3e primer to);
(6) primer formed of Nucleotide shown in the sequence 25 of Nucleotide and sequence table shown in the sequence 18 of sequence table is to (gluA3f primer to);
(7) primer formed of Nucleotide shown in the sequence 26 of Nucleotide and sequence table shown in the sequence 18 of sequence table is to (gluA3g primer to).
Said primer special also can be made up of said seven pairs of primers.
Said primer special can be applicable to prepare differentiates that wheat is the test kit that carries the wheat breed of which kind of Glu-A3 protein protomer.
It is the test kit that carries the wheat breed of which kind of Glu-A3 protein protomer that the present invention also protects a kind of wheat of differentiating, comprises described primer special.
The present invention also protects and a kind ofly differentiates that wheat to be measured is the method for carrying the wheat breed of which kind of Glu-A3 protein protomer, comprises the steps:
1) genomic dna with wheat to be measured is a template, carries out pcr amplification with said primer special;
2) detect pcr amplification product, judge that through the size of pcr amplification product wheat to be measured is the wheat breed that carries which kind of Glu-A3 protein protomer.
Judgement criteria is following: the genome with wheat to be measured is a template, adopts the gluA3a primer to carrying out PCR, if obtain the amplified production of 529bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3a.Genome with wheat to be measured is a template; Adopt the gluA3ac primer to carrying out PCR; If obtain the amplified production of 573bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3a or Glu-A3c, the PCR result associating right with the gluA3a primer; If use the Glu-A3a primer to not obtaining the amplified production of 529bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3c.Genome with wheat to be measured is a template, adopts the gluA3b primer to carrying out PCR, if obtain the amplified production of 894bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3b.Genome with wheat to be measured is a template, adopts the gluA3d primer to carrying out PCR, if obtain the amplified production of 967bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3d.Genome with wheat to be measured is a template, adopts the gluA3e primer to carrying out PCR, if obtain the amplified production of 158bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3e.Genome with wheat to be measured is a template, adopts the gluA3f primer to carrying out PCR, if obtain the amplified production of 552bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3f.Genome with wheat to be measured is a template, adopts the gluA3g primer to carrying out PCR, if obtain the amplified production of 1345bp, the Glu-A3 protein protomer of said wheat to be measured is Glu-A3g.
Said primer special, said test kit and said method all can be applicable to wheat breeding.
Said wheat specifically can be common wheat.
Utilize primer special of the present invention, test kit or method growing the early screening wheat breed, thereby optimize cross combination, accelerate breeding speed, reduce the breeding cost.Method provided by the invention is simple to operate, reliable and stable, with low cost, is suitable for applying, and is significant with the improvement wheat quality to research wheat quality correlated character.The present invention will play a significant role in Wheat Breeding for Quality.
Description of drawings
Fig. 1 is the pcr amplification detected result of gluA3a primer to 7 materials in the his-and-hers watches 1.
Fig. 2 is the pcr amplification detected result of gluA3b primer to 7 materials in the his-and-hers watches 1.
Fig. 3 is the pcr amplification detected result of gluA3ac primer to 7 materials in the his-and-hers watches 1.
Fig. 4 is the pcr amplification detected result of gluA3d primer to 7 materials in the his-and-hers watches 1.
Fig. 5 is the pcr amplification detected result of gluA3e primer to 7 materials in the his-and-hers watches 1.
Fig. 6 is the pcr amplification detected result of gluA3f primer to 7 materials in the his-and-hers watches 1.
Fig. 7 is the pcr amplification detected result of gluA3g primer to 7 materials in the his-and-hers watches 1.
Fig. 8 is SDS-PAGE test section material Glu-A3 subunit result; Wherein representes the purpose band.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment like no specified otherwise, is ordinary method.Used test materials among the following embodiment like no specified otherwise, is to buy from routine biochemistry reagent shop and obtains.Primer among the following embodiment is synthetic gives birth to worker's completion with dna sequencing by Shanghai.The wheat lines that all are used in following examples is all available from country of Chinese Academy of Agricultural Sciences farm crop quality saving center.
The acquisition of embodiment 1, common wheat low molecular weight glutenin Glu-A3 locus gene
Used wheat breed is seen table 1.
Table 1 is used to clone Glu-A3 locus gene and the wheat breed of analyzing Glu-A3 subunit specific mark
Seven wheat breeds in the table 1 are wheat (the Vawser MJ that contains definite Glu-A3 protein protomer; Cornish GB; Shepherd KW (2002) Rheological dough properties of Aroonaisolines differing in glutenin subunit composition.In:Cereals 2002; Proceedings of the 52nd Australian Cereal Chemistry Conference.Christchurch; New Zealand. (Eds CK Black, JF Panozzo, CW Wrigley; IL Bateyand N Larsen) pp.53-58. (Cereal Chemistry Division, Royal AustralianChemical Institute)).
Go up Glu-A3 site correlated series (GENBANK ACCESSION NO.EU189087, AB062868, FJ441117 and EF426565) according to GenBank, it is following to design 3 pairs of primers:
Primer is to first: P1 (upstream primer): 5 '-AAACAGAATTATTAAAGCCGG-3 ';
P2 (downstream primer): 5 '-CAGGCAAGACAAACTTATGAA-3 ';
Primer pair B:
P3 (upstream primer): 5 '-CATCTCTGATCCATCCAAACAT-3 ';
P4 (downstream primer): 5 '-GTGATCCAACTATATATTACTAGAC-3 ';
Primer is to third:
P5 (upstream primer): 5 '-TTCAGATGCAGCCAAACAA-3 ';
P6 (downstream primer): 5 '-CAAACTTATGAACTTGCAAGG-3 '.
Utilize the genomic dna of the wheat in above-mentioned 3 pairs of primer his-and-hers watches 1 to carry out pcr amplification respectively.
In the pcr amplification, the PCR reaction system of per 20 μ l is following: the template DNA of 50ng, Taq enzyme 1U (TaKaRa), upstream and downstream primer (5 μ molL
-1) each 1.2 μ l, dNTP (2.5mmolL
-1, TaKaRa) 0.8 μ l, 10 * PCR damping fluid, 2 μ l are with sterile distilled water postreaction system to 20 μ l.
PCR response procedures: 94 ℃ of 5min; 94 ℃ of 35s then, 57 ℃ (first and second pairs of primers) or 58 ℃ (the 3rd pair of primer) annealing 35s, 72 ℃ are extended 1min 30s, totally 38 circulations; Last 72 ℃ of 5min, 4 ℃ of insulations.
Primer obtains a Glu-A3 locus gene, called after GluA3-1 to the pcr amplification product order-checking back of first.GluA3-1 comprises 7 allelic variations (seeing the sequence 7 of sequence 1 to the sequence table of sequence table) altogether.
Obtained a Glu-A3 locus gene, called after GluA3-2 after the pcr amplification product order-checking of primer pair B.GluA3-2 comprises totally 4 allelic variations (seeing the sequence 11 of sequence 8 to the sequence table of sequence table) altogether.
Primer has obtained a Glu-A3 locus gene, called after GluA3-3 after to the order-checking of third pcr amplification product.GluA3-3 comprises 6 allelic variations (seeing the sequence 17 of sequence 12 to the sequence table of sequence table) altogether.
The acquisition of embodiment 2, common wheat low molecular weight glutenin Glu-A3 subunit specific mark
According to 3 Glu-A3 genes of above-mentioned acquisition and the corresponding relation between the Glu-A3 protein protomer, right behind the sequence alignment according to the Auele Specific Primers of 7 pairs of identification of the difference design Glu-A3 protein protomers between the allele variation, see table 2.
Table 2 differentiates that the Auele Specific Primer of Glu-A3 protein protomer is right
Utilize the genomic dna of the wheat in above-mentioned 7 pairs of primer his-and-hers watches 1 to carry out pcr amplification respectively.
The PCR reaction system is following: the template DNA of 50ng, Taq enzyme 1U (TaKaRa), upstream and downstream primer (5 μ molL
-1) each 1.2 μ l, dNTP (2.5mmolL
-1, TaKaRa) 0.8 μ l, 10 * PCR damping fluid, 2 μ l are with sterile distilled water postreaction system to 20 μ l.
PCR response procedures: 94 ℃ of 5min; 94 ℃ of 35s then, 60 ℃ of annealing 35s, 72 ℃ are extended 1min30s, totally 38 circulations; Last 72 ℃ of 5min, 4 ℃ of insulations.
After the PCR reaction finishes; Amplified production is carried out 1.5% agarose gel analysis, 200V voltage electrophoresis 20 minutes, (EB); 0.5% ethidium bromide (EB) dyeing is 10 minutes then, after the zero(ppm) water rinsing on Gel Doc XRSystem imaging system with the uv lamp scanning imagery and deposit computingmachine in.
The detected result that adopts gluA3a to carry out the product of pcr amplification is seen Fig. 1.Among Fig. 1, M: molecular weight standard; 1-7: sample 1-7.The wheat sample that contains the Glu-A3a protein protomer shows the special band of a 529bp.The detected result that adopts gluA3b to carry out the product of pcr amplification is seen Fig. 2.Among Fig. 2, M: molecular weight standard; 1-7: sample 1-7.The wheat sample that contains the Glu-A3b protein protomer shows the special band of a 894bp.The detected result that adopts gluA3ac to carry out the product of pcr amplification is seen Fig. 3.Among Fig. 3, M: molecular weight standard; 1-7: sample 1-7.The wheat sample standard deviation that contains Glu-A3a and Glu-A3c protein protomer shows the special band of a 573bp.The detected result that adopts gluA3d to carry out the product of pcr amplification is seen Fig. 4.Among Fig. 4, M: molecular weight standard; 1-7: sample 1-7.The wheat sample that contains the gluA3d protein protomer shows the special band of a 967bp.The detected result that adopts gluA3e to carry out the product of pcr amplification is seen Fig. 5.Among Fig. 5, M: molecular weight standard; 1-7: sample 1-7.The wheat sample that contains the gluA3e protein protomer shows the special band of a 158bp.The detected result that adopts gluA3f to carry out the product of pcr amplification is seen Fig. 6.Among Fig. 6, M: molecular weight standard; 1-7: sample 1-7.The wheat sample that contains the gluA3f protein protomer shows the special band of a 552bp.The detected result that adopts gluA3g to carry out the product of pcr amplification is seen Fig. 7.Among Fig. 7, M: molecular weight standard; 1-7: sample 1-7.The wheat sample that contains the gluA3g protein protomer shows the special band of a 1345bp.
For further verifying 7 pairs of practicality that Auele Specific Primer is right of exploitation, utilize 7 materials (numbering 1-7) that are used for clone gene among 121 parts of wheat lines of 7 pairs of primers detections (numbering 8-128) and the embodiment 1, the result sees table 3.The Glu-A3 site protein protomer of these materials is analyzed through SDS-PAGE, thereby can verify 7 pairs of detected results that Auele Specific Primer is right.Fig. 8 is SDS-PAGE test section material Glu-A3, Glu-B3 and Glu-D3 subunit result.Among Fig. 8,1:21; 2:26; 3:30; 4:Pav ó n (contrast); 5:8; 6:11; 7:17; 8:19; 9:31; 10:90; 11:18; 12:24; 13:25; 14:Opata (contrast); Above numeral all refers to the numbering in the table 3.Contrast is for the reference position of respective strap is provided.
The checking of the special STS mark of table 3Glu-A3 protein protomer
The result finds that in 128 parts of materials, the detected result of two kinds of methods is consistent.
Sequence table
< 110>Institute of Crop Science, Chinese Academy of Agricultural Science
< 120>differentiate that wheat is the primer special and the application thereof of carrying which kind of Glu-A3 protein protomer kind
<130>CGGNARY92315
<160>26
<210>1
<211>1501
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>1
aaacagaatt?attaaagccg?gtgcaaagaa?ggaaaagagg?tggtgtcccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?cacaagcatc?aaagccaaac?aacactagtt 120
aacaccaatc?cacaatgaag?accttcctcg?tctttgccct?cctcgctctt?gcggcggcaa 180
gtgccgttgc?gcaaatttca?cagcaacaac?aacaaccacc?attttcacag?caacaacaac 240
caccattttc?gcagcaacag?caatcaccat?tttcgcaaca?acaacaacaa?ccaccatttt 300
tgcagcaaca?acaaccaccg?ttttcacaac?agccaccaat?ttcacagcag?caacaaccac 360
cattttcaca?gcaacaacaa?ccacaatttt?cacagcaaca?gcagccacca?tattcgcagc 420
aacaacagcc?accatattcg?cagcaacaac?aaccaccatt?ttcgcagcaa?cagcaaccac 480
cattttcgca?gcaacaacaa?ccaccatttt?tgcagcaaca?acaacaacca?ccatttacac 540
agcaacaaca?accatcgttt?tcacaacagc?caccaatttc?acagcagcag?cagcagcaac 600
aacaacaaca?acaaccattt?acacagcaac?aacaaccacc?attttcacaa?cagccaccaa 660
tttcacagca?gcaacaacca?ccattttcgc?agcaacaaca?accaccattt?tcacagcaac 720
aacaaatacc?agttattcat?ccatctgttt?tgcagcagct?aaacccatgc?aaggtattcc 780
tccagcagca?gtgcatccct?gtggcaatgc?agcgatgtct?tgctaggtca?caaatgttgc 840
agcagagcat?ttgccatgtg?atgcagcaac?aatgttgcca?gcagttgcgg?caaatccccg 900
agcaatcccg?ccatgagtca?atccgtgcta?tcatctactc?tatcatcctg?cagcagcagc 960
agcagcaaca?acaacaacaa?caacagggtc?agagtatcat?ccaatatcag?caacaacaac 1020
cccaacagtt?gggccaatgt?gtctcccaac?cccaacagca?gttgcagcag?caactcgggc 1080
aacaacctca?acaacaacaa?ttggcacatg?gtaccttttt?gcagccacac?cagatagctc 1140
agcttgaggt?gatgacttcc?attgcactcc?gtaccctgcc?aacaatgtgc?agtgtcaatg 1200
tgccgttgta?cgaaaccacc?actagtgtgc?cattaggcgt?tggcatcgga?gttggtgtct 1260
actgataaga?aaagatctct?agtaatatat?agttggatca?ccgttgttta?gtcgatggat 1320
atgtcaatgt?agcggtgaca?aataaagtgt?cacacaacgt?catgtgtgac?ccgctcaaac 1380
tagttgttta?aattctgaaa?taaaatacaa?ataaaggtgt?attaagaaaa?tgttcatatc 1440
ggcattgtgt?ggatgtcgat?ctgattgcca?tgcatgcaag?ttcataagtt?tgtcttgcct 1500
g 1501
<210>2
<211>1525
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>2
aaacagaatt?attaaagccg?gtgcaaagaa?ggaaaagagg?tggtgtcccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?cacaagcatc?aaagccaaac?aacactagtt 120
aacaccaatc?cacaatgaag?accttcctcg?tctttgccct?cctcgctctt?gcggcggcaa 180
gtgccgttgc?gcaaatttca?cagcaacaac?aaccaccacc?attttcgcag?caacaacaac 240
caccattttc?acagcaacaa?caaccaccat?tttcgcagca?acagcaatca?ccattttcgc 300
aacaacaaca?acaaccacca?tttttgcagc?aacaacaacc?accgttttca?caacaaccac 360
caatttcaca?gcagcaacaa?ccaccatttt?cacagcaaca?acaaccacaa?ttttcacagc 420
aacagcagcc?accatattcg?cagcaacaac?aaccaccata?ttcgcagcaa?caacaaccac 480
cattttcgca?gcaacagcaa?ccaccatttt?cgcagcaaca?acaaccacca?ttttcgcagc 540
aacaacaaca?accaccattt?acacagcaac?aacaaccacc?gttttcacaa?cagccaccaa 600
tttcacagca?gcagcaacag?caacaacaac?aacaacaacc?atttacacag?caacaacaac 660
caccgttttc?acaacagcca?ccaatttcac?agcagcaaca?accaccattt?tcgcagcaac 720
aacaaccacc?attttcacag?caacaacaaa?taccagttat?tcatccatct?gttttgcagc 780
agctaaaccc?atgcaaggta?ttcctccagc?agcagtgcat?ccctgtggca?atgcagcgat 840
gtcttgctag?gtcacaaatg?ttgcagcaga?gcatttgcca?tgtgatgcag?caacaatgtt 900
gccagcagtt?gcggcaaatc?cccgagcaat?cccgccatga?gtcaatccgt?gctatcatct 960
actctatcat?cctgcagcag?cagcagcagc?aacaacaaca?acaacaacag?ggtcagagta 1020
tcatccaata?tcagcaacaa?caaccccaac?agttgggcca?atgtgtctcc?caaccccaac 1080
agcagttgca?gcagcaactc?gggcaacaac?ctcaacaaca?acaattggca?catggtacct 1140
ttttgcagcc?acaccagata?gctcagcttg?aggtgatgac?ttccattgca?ctccgtaccc 1200
tgccaacaat?gtgcagtgtc?aatgtgccgt?tgtacgaaac?caccactagt?gtgccattag 1260
gcgttggcat?cggagttggt?gtctactgat?aagaaaagat?ctctagtaat?atatagttgg 1320
atcaccgttg?tttagtcgat?ggatatgtca?atgtagcggt?gacaaataaa?gtgtcacaca 1380
acgtcatgtg?tgacccgctc?aaactagttg?tttaaattct?gaaataaaat?acaaataaag 1440
gtgtattaag?aaaatgttca?tatcggcatt?gtgtggatgt?cgatctgatt?gccatgcatg 1500
caagttcata?agtttgtctt?gcctg 1525
<210>3
<211>1501
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>3
aaacagaatt?attaaagccg?gtgcaaagaa?ggaaaagagg?tggtgtcccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?cacaagcatc?aaagccaaac?aacactagtt 120
aacaccaatc?cacaatgaag?accttcctcg?tctttgccct?cctcgctctt?gcggcggcaa 180
gtgccgttgc?gcaaatttca?cagcaacaac?aacaaccacc?attttcacag?caacaacaac 240
caccattttc?gcagcaacag?caatcaccat?tttcgcaaca?acaacaacaa?ccaccatttt 300
tgcagcaaca?acaaccaccg?ttttcacaac?agccaccaat?ttcacagcag?caacaaccac 360
cattttcaca?gcaacaacaa?ccacaatttt?cacagcaaca?gcagccacca?tattcgcagc 420
aacaacagcc?accatattcg?cagcaacaac?aaccaccatt?ttcgcagcaa?cagcaaccac 480
cattttcgca?gcaacaacaa?ccaccatttt?cgcagcaaca?acaacaacca?ccatttacac 540
agcaacaaca?accatcgttt?tcacaacagc?caccaatttc?acagcagcag?cagcagcaac 600
aacaacaaca?acaaccattt?acacagcaac?aacaaccacc?attttcacaa?cagccaccaa 660
tttcacagca?gcaacaacca?ccattttcgc?agcaacaaca?accaccattt?tcacagcaac 720
aacaaatacc?agttattcat?ccatctgttt?tgcagcagct?aaacccatgc?aaggtattcc 780
tccagcagca?gtgcatccct?gtggcaatgc?agcgatgtct?tgctaggtca?caaatgttgc 840
agcagagcat?ttgccatgtg?atgcagcaac?aatgttgcca?gcagttgcgg?caaatccccg 900
agcaatcccg?ccatgagtca?atccgtgcta?tcatctactc?tatcatcctg?cagcagcagc 960
agcagcaaca?acaacaacaa?caacagggtc?agagtatcat?ccaatatcag?caacaacaac 1020
cccaacagtt?gggccaatgt?gtctcccaac?cccaacagca?gttgcagcag?caactcgggc 1080
aacaacctca?acaacaacaa?ttggcacatg?gtaccttttt?gcagccacac?cagatagctc 1140
agcttgaggt?gatgacttcc?attgcactcc?gtaccctgcc?aacaatgtgc?agtgtcaatg 1200
tgccgttgta?cgaaaccacc?actagtgtgc?cattaggcgt?tggcatcgga?gttggtgtct 1260
actgataaga?aaagatctct?agtaatatat?agttggatca?ccgttgttta?gtcgatggat 1320
atgtcaatgt?agcggtgaca?aataaagtgt?cacacaacgt?catgtgtgac?ccgctcaaac 1380
tagttgttta?aattctgaaa?taaaatacaa?ataaaggtgt?attaagaaaa?tgttcatatc 1440
ggcattgtgt?ggatgtcgat?ctgattgcca?tgcatgcaag?ttcataagtt?tgtcttgcct 1500
g 1501
<210>4
<211>1443
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>4
aaacagaatt?attaaagccg?gtgcaaaaaa?ggaaaagagg?tggtgttccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?cacaagcatc?aaagccaagc?aacactagtt 120
aacaccaatc?cacaatgaag?accttcctcg?tctttgccct?cctcgctctt?gcggcggcga 180
gtgccgttgc?gcaaatttca?cagcaacaac?aaccaccact?attttcgcag?caacaacaac 240
caccattttc?acagcaacaa?caaccaccat?tttcgcagca?acagcaatca?ccattttcgc 300
aacaacaaca?accaccattt?tcgcagcaac?aacaaccacc?gttttcacaa?caaccaccaa 360
tttcacagca?gcaacaacca?caatttttgc?agcaacaaca?accaccattt?tcacagcaac 420
agcaaccacc?attttcgcag?caacaacagc?caccatattc?gcagcaacaa?caaccaccat 480
tttcacagca?acaacaacca?ccattttcgc?agcaacaaca?accaccattt?tcgcagcagc 540
aacaacaaca?acaaccacca?tttacacaac?aacaacaacc?accgttttca?caacaaccac 600
caatttcaca?gcagcaacaa?ccaccatttt?cgcagcaaca?acaaccacca?ttttcacagc 660
aacaacaaat?accagttatt?catccatctg?ttctgcagca?gctaaaccca?tgcaaggtat 720
tcctccagca?gcagtgcatc?cctgtggcaa?tgcagcgatg?tcttgctagg?tcacaaatgt 780
tgcagcagag?catttgccat?gtgatgcagc?aacaatgttg?ccagcaattg?cggcaaatcc 840
ccgagcaatc?ccgccatgag?tcaatccgtg?ctatcgtcta?ctctatcatc?ctgcagaagc 900
aacaacaaca?acaacaacaa?caacagggtc?agagtatcat?ccaatatcag?caacaacaac 960
cccaacagtt?gggccaatgt?gtctcccaac?cccaacagca?gttgcagcag?caactcgggc 1020
aacaacctca?acaacaacaa?ttgacacatg?gtgccttttt?gcagccacac?cagatagctc 1080
agcttgaggt?gatgaattcc?attgcactcc?gtaacctgcc?aaggatgtgc?agtgtcaacg 1140
tgccgttgta?cgaaaccacc?actagtgtgc?cattaggcgt?tggcatcgga?gttggtgtct 1200
actgataaga?aaagatctct?agtaatatat?agttggatca?ccgttgttta?gtcgatggat 1260
atgtcgatgc?agcggtgaca?aataatgtgt?cacacaacgt?catgtgtgac?ccgctcaaac 1320
tagttgttta?aattctgaaa?taaaatacaa?ataaagttgt?atcaagacaa?tgttcatatc 1380
ggcattgtgt?ggatatgtcg?atctgattgc?catgcttgca?ggttcataag?tttgtcttgc 1440
ctg 1443
<210>5
<211>1540
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>5
aaacagaatt?attaaagccg?gtgcaaagaa?ggaaaagagg?tggtgtcccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?cacaagcatc?aaagccaagc?aacactagtt 120
aacaccaatc?cacaatgaaa?accttcctcg?tctgtgccct?cctcgctctt?gcggcggcaa 180
gtgccgttgc?gcaaatttca?cagcaacaac?aaccaccacc?attttcgcag?caacaacaac 240
caccattttc?acagcaacaa?caaccaccat?tttcgcagca?acaacaatca?ccattttcgc 300
aacaacaaca?acaaccacca?ttttcgcagc?aacaacaacc?accattttca?caacaaccac 360
taatttcaca?gcagcaacaa?ctaccatttt?cacagcaaca?acaaccacaa?ttttcacagc 420
aacagcaacc?accatattcg?cagcaacaac?agccaccata?ttcgcagcaa?caacaaccac 480
cattttcgca?gcaacagcaa?ccaccatttt?cgcagcaaca?acaaccatca?ttttcgcagc 540
aacaacaaca?accaccattt?acacaacaac?aacaaccacc?gttttcacaa?cagtcaccaa 600
tttcacagca?gcagcaacaa?caacaacaac?aacaacaaca?accatttaca?cagcaacagc 660
aaccaccgtt?ttcacaacag?ccaccaattt?cacagcagca?acaaccacca?ttttcgcagc 720
aacaacaacc?accattttca?cagcaacaac?aaataccagt?tattcatcca?tctgttttgc 780
agcagctaaa?cccatgcatg?gtattcctcc?agcagcagtg?catccctgtg?gcaatgcagc 840
gatgtcttgc?taggtcacaa?atgttgcagc?agagcatttg?ccatgtgatg?cagcgacaat 900
gttgccagca?gttgcggcaa?atccccgagc?aatcccgcca?tgagtcaatc?cgtgctatca 960
tctactctat?catcctgcag?cagcagcagc?aacaacaaca?acaacaacaa?caacaacaac 1020
aacagggtca?gagtatcatc?caatatcagc?aacaaccacc?ccaacagttg?ggccaatgtg 1080
tctcccaacc?ccaacagcag?ttgcagcagc?aactcgggca?acaacctcaa?caacaacagt 1140
tggcacatgg?tacctttttg?cagccacacc?agatagctca?acttgaggtg?atgacttcca 1200
ttgcactccg?taccctgcca?agaatgtgca?gtgtcaatgt?gccgttgtac?gaaaccacca 1260
ctagtgtgcc?attaggcgtt?ggcatcggag?ttggtgtcta?ctgataagaa?aagatctcta 1320
gtaatatata?gttggatcac?ggttgtttag?tcgatggata?tgtcgatgta?gcggtgacaa 1380
ataaagtgtc?acacaacgtc?atgtgtgacc?cactcaaaat?agttgtttaa?attctgaaat 1440
aaaatacaaa?taaaggtgta?ttaagaaaat?gttcatatcg?gcattgtgtg?gatgtcgatc 1500
tgattgccat?gcatgcaagt?tcataagttt?gtcttgcctg 1540
<210>6
<211>1441
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>6
aaacagaatt?attaaagccg?gtgcaaagaa?ggaaaagagg?tggtgtcccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?tacaagcatc?aaagccaagc?aacactagtt 120
aacaccaatc?cacaatgaag?accttcctcg?tctttgccct?cctcgctctt?gcggcggcaa 180
gtgccgttgc?gcaaatttca?cagcaacaac?aagcaccgcc?attttcgcag?caacaacaac 240
caccattttc?acagcaacaa?caaccaccat?tttcgcagca?acaacaatca?ccattttcgc 300
aacaacaaca?acaaccacca?tttgcgcagc?aacaacaacc?accgttttca?caacaaccac 360
caatttcaca?gcagcaacaa?ccaccatttt?cacagcaaca?acaaccacaa?ttttcacagc 420
aacaacaacc?accatattcg?cagcaacaac?agccaccata?ttcgcagcaa?caacaaccac 480
cattttcgca?gcaacaacaa?ccaccatttt?cgcagcaaca?acaacaacca?ccatttacac 540
agcagcagca?gcagcaacaa?caacaacaac?catttacaca?gcaacagcaa?ccaccgtttt 600
cacaacagcc?accaatttca?cagcagcaac?aaccaccatt?tttgcagcaa?caacgaccac 660
cattttcacg?gcaacaacaa?ataccagtta?ttcatccatc?tgttttgcaa?cagctaaacc 720
catgcaaggt?attcctccaa?cagcagtgca?tccctgtggc?aatgcagcga?tgtcttgcta 780
ggtcacaaat?gttgcagcag?agcatttgcc?atgtgatgca?gcaacaatgt?tgccagcagt 840
tgcggcaaat?ccccgagcaa?tcccgccatg?agtcaatccg?tgctatcatc?tactctatca 900
tcctgcagca?gcagcagcag?caacaacaac?aacaacaaca?acaacagggt?cagagtatca 960
tccaatatca?gcaacaacaa?ccccaacagt?tgggccaatg?tgtctcccaa?cccctacagc 1020
agttgcagca?gcaactcggg?caacaacctc?aacaacaaca?attggcacac?cagatagctc 1080
agcttgaggt?gatgacttcc?attgcactcc?gtaccctgcc?aacaatgtgc?aatgtcaatg 1140
tgccgttgta?cgaaaccacc?actagtgtgc?cattaggcgt?tggcatcgga?gttggtgtct 1200
actgataaga?aaagatctct?agtaatatat?agttggatca?ccgttgttta?gtcgatggat 1260
atgtcgatgt?agcggtgaca?aataaagtgt?cacacaacgt?catgtgtgac?ccgctcaaac 1320
tagttgttta?aattctgaaa?taaaatacaa?ataaaggtgt?attaagaaaa?tgttcatatc 1380
ggcattgtgt?ggatgtcgat?ctgattgcca?tgcatgcaag?ttcataagtt?tgtcttgcct 1440
g 1441
<210>7
<211>1537
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>7
aaacagaatt?attaaagccg?gtgcaaagaa?ggaaaagagg?tggtgtcccg?gcaactataa 60
ataggcatga?agtataaaga?tcatcacaag?cacaagcatc?aaagccaagc?aacactagtt 120
aacaccaatc?cacaatgaag?accttcctcg?tctttgccct?cctcgctctt?gcggcggcaa 180
gtgccgttgc?gcaaatttca?cagcaacaac?aaccaccacc?attttcgcag?caacaacaac 240
caccattttc?acagcaacaa?caaccaccat?tttcgcagca?acaacaatca?ccattttcgc 300
aacaacaaca?acaaccacca?ttttcgcagc?aacaacaacc?accattttca?caacaaccac 360
taatttcaca?gcagcaacaa?ctaccatttt?cacagcaaca?acaaccacaa?ttttcacagc 420
aacagcagcc?accatattcg?caacaacaac?agccaccata?ttcgcagcaa?caacaaccac 480
cattttcgca?gcaacagcaa?ccaccatttt?cgcagcaaca?acaaccatca?ttttcgcagc 540
aacaacaaca?accaccattt?acacagcaac?aacaaccacc?gttttcacaa?cagtcaccaa 600
tttcacagca?gcagcagcaa?caacaacaac?aacaacaaca?accatttaca?cagcaacagc 660
aaccaccgtt?ttcacaacag?ccaccaattt?cacagcagca?acaaccacca?ttttcgcagc 720
aacaacaacc?accattttca?cagcaacaac?aaataccagt?tattcatcca?tctgttttgc 780
agcagctaaa?cccatgcaag?gtattcctcc?agcagcagtg?catccctgtg?gcaatgcagc 840
gatgtcttgc?taggtcacaa?atgttgcagc?agagcatttg?ccatgtgatg?cagcgacaat 900
gttgccagca?gttgcggcaa?atccccgagc?aatcccgcca?tgagtcaatc?cgtgctatca 960
tctactctat?catcctgcag?cagcagcagc?agcaacaaca?acaacaacaa?caacaacaac 1020
agggtcagag?tatcatccaa?tatcagcaac?aaccacccca?acagttgggc?caatgtgtct 1080
cccaacccca?acagcagttg?cagcagcaac?tcgggcaaca?acctcaacaa?caacagttgg 1140
cacatggtac?ctttttgcag?ccacaccaga?tagctcagct?tgaggtgatg?acttccattg 1200
cactccgtac?cctgccaaga?atgtgcagtg?tcaatgtgcc?gttgtacgaa?accaccacta 1260
gtgtgccatt?aggcgttggc?atcggagttg?gtgtctactg?ataagaaaag?atctctagta 1320
atatttagtt?ggatcaccgt?tgtttagtcg?atggatatgt?cgatgtagcg?gtgacaaata 1380
aagtgtcaca?caacgtcatg?tgtgacccac?tcaaactagt?tgtttaaatt?ctgaaataaa 1440
atacaaataa?aggtgtatta?agaaaatgtt?catatcggca?ttgtgtggat?gtcgatctga 1500
ttgccatgca?tgcaagttca?taagtttgtc?ttgcctg 1537
<210>8
<211>1100
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>8
catctctgat?ccatccaaac?ataattatta?aagttgatgc?aaagaaggaa?agaggtggtg 60
cccggactac?tataaatagg?catgaagtat?aaagatcatc?acaagcacaa?gcatcaaaac 120
caagcaaaac?tagttagcac?caatccatca?tgaagacctt?cctcgtcttt?gccctcctcg 180
ccgttgtggc?aacaagtgcc?attgcgcaga?tggatactag?ctacatccct?ggtttggaga 240
gaccatggca?gcagcaacca?ttaccaccac?aacagacatt?tccacaacaa?ccaccatttt 300
cacaataaca?acaacaacaa?caacaacaac?catttcctca?acaaccatca?ttttcacagc 360
aacaaccacc?attttcacag?caacaaccaa?ttctaccaca?gggaccacca?ttttcacagc 420
aaacacaacc?tgttctaccg?caacaatcac?cattttcaca?gcaacaacaa?ctaattttac 480
ctccacaaca?acaacaacag?cttccgcaac?aacaaatctc?tattgttcaa?ccatccgttt 540
tgcagcagct?aaacccatgc?aaggtattcc?tccagcagca?gtgcagccct?gtggcaatgc 600
cacaacgtct?tgctaggtcg?caaatgtggc?agcagagccg?ttgccatgtg?atgcagcaac 660
aatgttgcca?gcagttgtcg?caaatccccg?aacaatcccg?ctatgatgca?atccgtgcca 720
tcacctactc?catcatccta?caagaacaac?aacagggttt?tgtccaagct?cagcagcaac 780
aaccccaaca?gtcaggtcaa?ggtgtctccc?aatcccaaca?gcagtcgcag?cagcagctcg 840
gacaatgttc?tttccaacaa?cctcaacagc?aactgggtca?acagcctcaa?caacaacagg 900
tacaacaggg?tacctttttg?cagccacacc?agatagctca?ccttgaggtg?atgacttcca 960
ttgcgctccg?taccctgcca?acgatgtgca?gtgtcaatgt?gccattgtac?agctccacca 1020
ctagtgtgcc?attcggagtt?ggcaccggag?ttggtgccta?ctgataagaa?aagatgtcta 1080
gtaatatata?gttggatcac 1100
<210>9
<211>1097
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>9
catctctgat?ccatccaaac?ataattatta?aagttgatgc?aaagaaggaa?agaggtggtg 60
cccggactac?tataaatagg?catgaagtat?aaagatcatc?acaagcacaa?gcatcaaaac 120
caagcaaaac?tagttagcac?caatccatca?tgaagacctt?cctcgtcttt?gccctcctcg 180
ccgttgtggc?aacaagtgcc?attgcgcaga?tggatactag?ctacatccct?ggtttggaga 240
gaccatggca?gcagcaacca?ttaccaccac?aacagacatt?tccacaacaa?ccaccatttt 300
cacaataaca?acaacaacaa?caacaaccat?ttcctcaaca?accatcattt?tcacagcaac 360
aaccaccatt?ttcacagcaa?caaccaattc?taccacaggg?accaccattt?tcacagcaaa 420
cacaacctgt?tctaccgcaa?caatcaccat?tttcacagca?acaacaacta?attttacctc 480
cacaacaaca?acaacagctt?ccgcaacaac?aaatctctat?tgttcaacca?tccgttttgc 540
agcagctaaa?cccatgcaag?gtattcctcc?agcagcagtg?cagccctgtg?gcaatgccac 600
aacgtcttgc?taggtcgcaa?atgtggcagc?agagccgttg?ccatgtgatg?cagcaacaat 660
gttgccagca?gttgtcgcaa?atccccgaac?aatcccgcta?tgatgcaatc?cgtgccatca 720
cctactccat?catcctacaa?gaacaacaac?agggttttgt?ccaagctcag?cagcaacaac 780
cccaacagtc?aggtcaaggt?gtctcccaat?cccaacagca?gtcgcagcag?cagctcggac 840
aatgttcttt?ccaacaacct?caacagcaac?tgggtcaaca?gcctcaacaa?caacaggtac 900
aacagggtac?ctttttgcag?ccacaccaga?tagctcacct?tgaggtgatg?acttccattg 960
cgctccgtac?cctgccaacg?atgtgcagtg?tcaatgtgcc?attgtacagc?tccaccacta 1020
gtgtgccatt?cggagttggc?accggagttg?gtgcctactg?ataagaaaag?atgtctagta 1080
atatatagtt?ggatcac 1097
<210>10
<211>1100
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>10
catctctgat?ccatccaaac?ataattatta?aagttgatgc?aaagaaggaa?agaggtggtg 60
cccggactac?tataaatagg?catgaagtat?aaagatcatc?acaagcacaa?gcatcaaaac 120
caagcaaaac?tagttagcac?caatccatca?tgaagacctt?cctcgtcttt?gccctccttg 180
ccgttgtggc?aacaagtgcc?attgcgcaga?tggatactag?ctgcatccct?ggtttggaga 240
gaccatggca?gcagcaacca?ttaccaccac?aacagacatt?tccacaacaa?ccaccatttt 300
cacaacaaca?acaacaacaa?caacaacaac?catttcctca?acaaccatca?ttttcacagc 360
aacaaccacc?attttcacag?caacaaccaa?ttctaccaca?gggaccacca?ttttcacagc 420
aaacacaacc?tgttctaccg?caacaatcac?cattttcaca?gcaacaacaa?ctaattttac 480
ctccacaaca?acaacaacag?cttccgcaac?aacaaatctc?tattgttcaa?ccatccattt 540
tgcagcagct?aaacccatgc?aaggtattcc?tccagcagca?gtgcagccct?gtggtaatgc 600
cacaacgtct?tgctaggtcg?caaatgtggc?agcagagcag?ttgccatgtg?atgcagcaac 660
aatgttgcca?gcagttgtcg?caaatccccg?aacaatcccg?ctatgatgca?atccgtgcca 720
tcacctaccc?catcatccta?caagaacaac?aacagggttt?tgtccaagct?cagcagcaac 780
aaccccaaca?gtcaggtcaa?ggtgtctccc?aatcccaaca?gcagtcgcag?cagcagctcg 840
gacaatgttc?tttccaacaa?cctcaacagc?aactgggtca?acagcctcaa?caacaacagg 900
tacaacaggg?tacctttttg?cagccacacc?agatagctca?tcttgaggtg?atgacttcca 960
ttgcgctccg?taccctgcca?acgatgtgca?gtgtcaatgt?gccattgtac?agctccacca 1020
ctagtgtgcc?attcggagtt?ggcaccggag?ttggtgccta?ctgataagaa?aagatgtcta 1080
gtaatatata?gttggatcac 1100
<210>11
<211>1106
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>11
catctctgat?ccatccaaac?ataattatta?aagttgatgc?aaagaaggaa?agaggtggtg 60
cccggactac?tataaatagg?catgaagtat?aaagatcatc?acaagcacaa?gcatcaaaac 120
caagcaaaac?tagttagcac?caatccatca?tgaagacctt?cctcgtcttt?gccctcctcg 180
ccattgtggc?aacaagtgcc?attgcgcaga?tggatactag?ctgcatccct?ggtttggaga 240
gacaatggca?gcagcaacca?ttaccaccac?aacagacatt?tccacaacaa?ccaccatttt 300
cacaacaaca?acaacaacaa?caacaacaac?aacaaccatt?tcctcaacaa?acatcatttt 360
cacagcaaca?accaccattt?tcacagcaac?aaccaattct?accacaggga?ccaccatttt 420
cacagcaaac?acaacctgtt?ataccgcaac?aatcaccatt?ttcacagcaa?caacaactaa 480
ttttacctcc?acaacaacaa?caacagcttc?cgcaacaaca?aatctctatt?gttcaaccat 540
ccgttttgca?gcagctaaac?ccatgcaagg?tattcctcca?gcagcagtgc?agccctgtgg 600
caatgccaca?acgtcttgct?aggtcgcaaa?tgtggcagca?gagcagttgc?catgtgatgc 660
agcaacaatg?ttgccagcag?ttgtcgcaaa?tccccgaaca?atcccgctat?gatgcaatcc 720
gtgccatcac?ctactccatc?atcctacaag?aacaacaaca?gggttttgtc?caagctcagc 780
agcaacaacc?ccaacagtca?ggtcaaggtg?tctcccaatc?ccaacagcag?tcgcagcagc 840
agctcggaca?atgttctttc?caacaacctc?aacagcaact?gggtcaacag?cctcaacaac 900
aacaggtaca?acagggtacc?tttttgcagc?cacaccagat?agctcacctt?gaggtgatga 960
cttccattgc?gctccgtacc?ctgccaatga?tgtgcagtgt?caatgtgcca?ttgtacagct 1020
ccaccactag?tgtgccattc?ggagttggca?ccggagttgg?tgcctactga?taagaaaaga 1080
tgtctagtaa?tatatagttg?gatcac 1106
<210>12
<211>1372
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>12
ttcagatgca?gccaaacaaa?tttattaaag?ctgatgcaaa?gaagaaaaga?ggtggtgccc 60
gggctactat?aaataggcat?gaagtataaa?gatcatcaca?agcacaagca?tcaaaaccaa 120
gcaacactag?ttgacatcaa?tccacaatga?agacattcct?cgtcttttcc?ctcctcgctc 180
ttgcggcggc?aagtgccgtt?gcgcaaattt?cacagcaaca?acaacaacaa?caaccatttc 240
cgcagcaaca?acaaccacca?ttttcgcagc?aacaagagcc?accatattca?cagcaatagc 300
aaccaccatt?ttcgcaacaa?caacaatcac?cattttcgca?gcaacaacaa?caacaaccac 360
catttttgca?gcaacaccaa?ccaccgtttt?cacaacagcc?accaatttca?cagcagcaac 420
aaccaccatt?ttcacagcaa?caacaaccac?cattttcaca?gcaacaacaa?ccaccatttt 480
cgcagcagca?acaacaacaa?ccaccatttt?cacaacaaca?acaaccaccg?ttttcacaac 540
agccaccaat?ttcacagcaa?caacaaccac?cattttcgca?gcaacaacaa?ccaccatttt 600
cacagcaaca?acaaatacca?gttattcatc?catctgtttt?gcagcaacta?aacccatgca 660
aggtattcct?ccagcagcag?tgcatccctg?tggcaatgca?gcgatgtctt?gctaggtcac 720
aaatgttgca?gcagaggatt?tgccatgtga?tgcagcaaca?atgttgccaa?cagttgcggc 780
aaatccccgg?gcaatcccgc?catgagtcaa?tccgtgctat?catctactct?atcatcctgc 840
agcaacaaca?acaacaacaa?caacaagttc?agagtatcat?ccaagctcag?caacaacaac 900
cccaacagtt?gggccaatgt?gtctcccaac?cccaacaaca?atcgcagcag?caactcgggc 960
aacaacctca?acaacaacaa?ttggcacagg?gtaccttttt?gcagccacac?cagatagctc 1020
aatttgaggt?gatgacttcc?attgcactcc?gtaccttgcc?aacgatgtgc?aatgtcaacg 1080
tgccgttgta?cggaaccacc?actagtgcgc?cattcggcgt?tggcactgga?gttggtgcct 1140
actgataaga?aaagatctct?attaatatat?agttggatca?ccgttgttta?gtcgatggat 1200
atgtcgatgt?agcggtgaca?aataaagtgt?cacacaacac?catgtgtgac?ccgcccaaac 1260
tagttgttta?aattctgaaa?taaaatacaa?ataaagttgt?atcaagacaa?tgttcatatc 1320
agcattgtgc?ggatgtcgat?ctgattgcca?tccttgcaag?ttcataagtt?tg 1372
<210>13
<211>1375
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>13
ttcagatgca?gccaaacaaa?tttattaaag?ctgatgcaaa?gaagaaaaga?ggtggtgccc 60
gggctactat?aaataggcct?gaagtataaa?gatcatcaca?agcacaagca?tcaaaaccaa 120
gcaacactag?ttgacaccaa?tccacaatga?agacattcct?cgtcttttcc?ctcctcgctc 180
ttgcggcggc?aagtgccgtt?gcgcaaattt?cacagcaaca?acaacaacaa?caaccatttc 240
cgcagcaaca?acaaccacca?ttttcgcagc?aacaagagcc?accatattca?cagcaatagc 300
aaccaccatt?ttcgcaacaa?caacaatcac?cattttcgca?gcaacaacaa?caacaaccac 360
catttttgca?gcaacaccaa?ccaccgtttt?cacaacagcc?accaatttca?cagcagcaac 420
aaccaccatt?ttcacagcaa?caacaaccac?cattttcaca?gcaacaacaa?ccaccatttt 480
cgcagcagca?acaacaacaa?ccaccatttt?cacaacaaca?acaaccaccg?ttttcacaac 540
agccaccaat?ttcacagcag?caacaaccac?cattttcgca?gcaacaacaa?ccaccatttt 600
cacagcaaca?acaaatacca?gttattcatc?catctgtttt?gcagcaacta?aacccataca 660
aggtattcct?ccagcagcag?tgcatccctg?tggcaatgca?gcgatgtctt?gctaggtcac 720
aaatgttgca?gcagaggatt?tgccatgtga?tgcagcaaca?atgttgccaa?cagttgcggc 780
aaatccccgg?gcaatcccgc?catgagttaa?tccgtgctat?catctactct?atcatcctgc 840
agcaacaaca?acaacaacaa?caacaacaag?tttagagtat?catccaagct?cagcgacaac 900
aaccccaaca?gttgggccaa?tgtgtctccc?aaccccaaca?acaatcgcag?cagcaactcg 960
ggcaacaacc?tcaacaacaa?caattggcac?agggtacctt?tttgcagcca?caccagatag 1020
ctcaatttga?ggtgatgact?tccattgcac?tccgtacctt?gccaacgatg?tgcaatgtca 1080
acgtgctgtt?gtacggaacc?accactagtg?cgccattcgg?cgttggcact?ggagttggtg 1140
cctactgata?agaaaagatc?tctattaata?tatagttgga?tcaccgttgt?ttagtcgatg 1200
gatatgtcga?tgtagcggtg?acaaataaag?tgtcacacaa?caccatgtgt?gacccgccca 1260
aactagttgt?ttaaattctg?aaataaaata?caaataaagt?tgtatcaaga?caatgttcat 1320
atcagcattg?tgcggatgtc?gatctgattg?ccatccttgc?aagttcataa?gtttg 1375
<210>14
<211>1405
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>14
ttcagatgca?gccaaacaaa?tttattaaag?ctgatgcaaa?gaagaaaaga?ggtggtggtc 60
gggctactat?aaataggcat?gaagtataaa?gatcatcaca?agcacaagca?tcaaaaccaa 120
gcaacactag?ttaacaccaa?tccacaatga?ggaccttcct?cgtctttgcc?ctcctcgctc 180
ttgcggcggc?aagtgtggtt?gtgcaaattt?cacagcaaca?acaacaacaa?ccatttccgc 240
agcaacaaca?accaccattt?tcgcagcaac?aacagccacc?attttcacaa?caacaacaac 300
caccattttc?gcaacaacaa?caatcaccat?tttcacagca?acaacaacaa?caaccacctt 360
ttttgcagca?acaacaacca?tcattttcac?aacagccacc?aatttcacag?cagcaacaac 420
caccattttc?gcagcaacaa?caaccaccat?tttcacagca?acaacaacca?ccattttcgc 480
agcagcaaca?acaacaacaa?caacaaccac?cattttcaca?acagccacca?atttcacagc 540
agcaacaacc?accatttgtg?cagcaacaac?aaccaccatt?ttcacagcaa?caacaaatac 600
cagttattca?tccatctgtt?ttgcagcaac?taaacccatg?caaggtattc?ctccagcagc 660
agtgcatccc?tgtggcaatg?cagcgatgtc?ttgctaggtc?acaaatgttg?cagcagagga 720
tttgccatgt?gatgcagcaa?caatgttgcc?agcagttgcg?gcaaatcccc?gagcaatccc 780
gccatgagtc?aattcgtgct?atcatctact?ctatcatcct?gcaacaacaa?caacaacaac 840
aacaacaaca?acaacaacaa?caacaagaac?aacaacaaca?acaacaacaa?catcaacaag 900
ttcagagtat?catccaagct?cagcaacaac?aaccccaaca?gttgggccta?tgtgtctccc 960
aaccccaaca?acagtcgcag?cagcaactcg?ggcaacaacc?tcaacaacaa?caattggcac 1020
agggtacctt?tttgcagcca?cactagatag?ctcagtttga?ggtgatgact?tccattgcgc 1080
tccgtacctt?gccaacgatg?tgcaatgtca?acgtgccgtt?gtacggaacc?accactagtg 1140
cgccattcgg?tgttggcgct?ggagttggtg?cctactgata?agaaaagatc?tcttgtaata 1200
tatagttgga?tcaccgttgt?ttagtcgatg?gatatgtcga?tgtagttgtg?acaaataaag 1260
tgtcacacaa?caccatgtgt?gacccagcca?aactagttgt?ttaaattctg?aaataaaata 1320
caaataaagt?tgtatcaaga?caatgttcat?atcagtattg?tgcggatgtc?gatctgattg 1380
ccatccttgc?aagttcataa?gtttg 1405
<210>15
<211>1381
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>15
ttcagatgca?gccaaacaaa?tttattaaag?ctgatgcaaa?gaagaaaaga?ggtggtgccc 60
gggctactat?aaataggcat?gaagtataaa?gatcatcaca?agcacaagca?tcaaaaccaa 120
gcaacactag?ttgacaccaa?tccacaatga?agacattcct?tgtctttgcc?ctcctcgctc 180
ttgcggcggc?aagtgccgtt?gcgcaaattt?cacagcaaca?acaacaacaa?ccatttccgc 240
ggcaacaaca?accaccattt?tcgcagcaac?aagagccacc?atattcacag?caacaacaac 300
caccattttt?gcaacaacaa?caatcaccat?tttagcagca?acaacaacaa?caaccaccat 360
ttttgcagca?acaccaacca?ccgttttcac?aacagccacc?aatttcacag?cagcaacaac 420
caccattttc?gcagcaacaa?caaccaccat?tttcacagca?acaacaacca?ccattttcgc 480
agcagcagca?acaacaacaa?ccaccatttt?cacagaaaca?acaaccatcg?ttttcacaac 540
agccaccaat?ttcacagcag?caacaaccac?cattttcgca?gcaacaacaa?ccaccatttt 600
cacagcaaca?acaaatacca?gttattcatc?catctgtttt?gcagcaacta?aacccatgca 660
aggtattcct?ccagcagcag?tgcatccctg?tggcaatgca?gcgatgtctt?gctaggtcgc 720
aaatgttgca?gcaaaggatt?tgccatgtga?tgcagcaaca?atgttgccaa?cagttgcggc 780
aaatccccgg?gcaatcccgc?catgagtcaa?tccgtgctat?catctactct?atcatcctgc 840
aacaacaaca?acaacaacaa?caacaacaac?aacaagttca?gagtatcatc?caagctcagc 900
aacaacaacc?ccaacaattg?ggccaatgtg?tcttccaacc?ccaacaacaa?tcgcagcagc 960
aactcgggca?acaacctcaa?caacaacaat?tggcacaggg?tacctttttg?cagccacacc 1020
agatagctca?gtttgaggtg?atgacttcca?ttgcactccg?taccttgcca?acgatgtgca 1080
atgtcaacgt?gccgttgtac?ggaaccacca?ctagtgcgcc?attcggcgtt?ggcactggag 1140
ttggtgccta?ctgataagaa?aagatctcta?gtaatatata?gttggatcac?tgttgtttag 1200
tcgatggata?tgtcgatgta?gcggtgacaa?ataaagtgtc?acacaacacc?atgtgtgacc 1260
cgcccaaact?agttgtttaa?attctgaaat?aaaatacaaa?taaagttgta?tcaagacaat 1320
gttcatatca?gcattgtgcg?gatgtcgatc?tgattgccat?ccttgcaagt?tcataagttt 1380
g 1381
<210>16
<211>1381
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>16
ttcagatgca?gccaaacaaa?tttattaaag?ctgatgcaaa?gaagaaaaga?ggtggtgccc 60
gggctactat?aaataggact?gaagtataaa?gatcatcaca?agcacaagca?tcaaaaccaa 120
gcaacactag?ttgacaccaa?tccacaatga?agacattcct?cgtcttttcc?ctcctcgctc 180
ttgcggcggc?aagtgccgtt?gcgcaaattt?cacagcaaca?acaacaacaa?caaccatttc 240
cgcagcaaca?acaaccacca?ttttcgcagc?aacaagagcc?accatattca?cagcaatagc 300
aaccaccatt?ttcgcaacaa?caacaatcac?cattttcgca?gcaacaacaa?caacaaccac 360
catttttgca?gcaacaccaa?ccaccgtttt?cacaacagcc?accaatttca?cagcagcaac 420
aaccaccatt?ttcacagcaa?caacaaccac?cattttcaca?gcaacaacaa?ccaccatttt 480
cgcagcagca?acaacaacaa?ccaccatttt?cacaacaaca?acaaccaccg?ttttcacaac 540
agccaccaat?ttcacagcag?caacaaccac?cattttcgca?gcaacaacaa?ccaccatttt 600
cacagcaaca?acaaatacca?gttattcatc?catctgtttt?gcagcaacta?aacccatgca 660
aggtattcct?ccagcagcag?tgcatccctg?tggcaatgca?gcgatgtctt?gctaggtcac 720
aaatgttgca?gcagaggatt?tgccatgtga?tgcagcaaca?atgttgccaa?cagttgcggc 780
aaatccccgg?gcaatcccgc?catgagttaa?tccgtgctat?catctactct?atcatcctgc 840
agcaacaaca?acaacaacaa?caacaacaac?aacaagttca?gagtatcatc?caagctcagc 900
gacaacaacc?ccaacagttg?ggccaatgtg?tctcccaacc?ccaacaacaa?tcgcagcagc 960
aactcgggca?acaacctcaa?caacaacaat?tggcacaggg?tacctttttg?cagccacacc 1020
agatagctca?atttgaggtg?atgacttcca?ttgcactccg?taccttgcca?gcgatgtgca 1080
atgtcaacgt?gctgttgtac?ggaaccacca?ctagtgcgcc?attcggcgtt?ggcactggag 1140
ttggtgccta?ctgataagaa?aagatctcta?ttaatatata?gttggatcac?cgttgtttag 1200
tcgatggata?tgtcgatgta?gcggtgacaa?ataaagtgtc?acacaacacc?atgtgtgacc 1260
cgcccaaact?agttgtttaa?attctgaaat?aaaatacaaa?taaagttgta?tcaagacaat 1320
gttcatatca?gcattgtgcg?gatgtcgatc?tgattgccat?ccttgcaagt?tcataagttt 1380
g 1381
<210>17
<211>1387
<212>DNA
< 213>Triticum common wheat (Triticum aestivum L.)
<400>17
ttcagatgca?gccaaacaaa?tttattaaag?ctgatgcaaa?gaagaaaaga?ggtggtgccc 60
gggctactat?aaataggcat?gaagtataaa?gatcatcaca?agcacaagca?tcaaaaccaa 120
gcaacactag?ttgacaccaa?tccacaatga?agacattcct?tgtctttgcc?ctcctcgctc 180
ttgcggcagc?aagtgccgtt?gcgcaaattt?cacagcaaca?acaacaacaa?ccatttccgc 240
ggcaacaaca?accaccattt?ttgcagcaac?aagagccacc?atattcacag?caacagcaac 300
caccattttc?gcaacaacaa?caatcaccat?tttagcagca?acaacaacaa?caaccaccat 360
ttttgcagca?acaccaacca?ccgttttcac?aacagccacc?aatttcacag?cagcaacaac 420
caccattttc?gcagcaacaa?caaccaccat?tttcacagca?acaacaacca?ccattttcgc 480
agcagcaaca?acaacaacaa?ccaccatttt?cacagaaaca?acaaccatcg?ttttcacaac 540
agccaccaat?ttcacagcag?caacaaccac?cattttcgca?gcaacaacaa?ccaccatttt 600
cacagcaaca?acaaatacca?gttattcatc?catctgtttt?gcagcaacta?aacccatgca 660
aggtattcct?ccagcagcag?tgcatccctg?tggcaatgca?gcgatgtctt?gctaggtcgc 720
aaatgttgta?gcaaaggatt?tgccatgtga?tgcagcaaca?atgttgccaa?cagttgcggc 780
aaatccccgg?gcaatcccgc?catgagtcaa?tccgtgctat?catctactct?atcatcctgc 840
aacaacaaca?acaacaacaa?caacaacaac?aacaacaaca?agttcagagt?atcatccaag 900
ctcagcaaca?acaaccccaa?caattgggcc?aatgtgtctt?ccaaccccaa?caacaatcgc 960
agcagcaact?cgggcaacaa?cctcaacaac?aacaattggc?acagggtacc?tttttgcagc 1020
cacaccagat?agctcagttt?gaggtgatga?cttccattgc?actccgtacc?ttgccaacga 1080
tgtgcaatgt?caacgtgccg?ttgtacggaa?ccaccactag?tgcgccattc?ggcgttggca 1140
ctggagttgg?tgcctactga?taagaaaaga?tctctagtaa?tatatagttg?gatcactgtt 1200
gtttagtcga?tggatatgtc?gatgtagcgg?tgacaaataa?agtgtcacac?aacaccatgt 1260
gtgacccgcc?caaactagtt?gtttaaattc?tgaaataaaa?tacaaataaa?gttgtatcaa 1320
gacaatgttc?atatcagcat?tgtgcggatg?tcgatctgat?tgccatcctt?gcaagttcat 1380
aagtttg 1387
<210>18
<211>21
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>18
aaacagaatt?attaaagccg?g 21
<210>19
<211>19
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>19
ttcagatgca?gccaaacaa 19
<210>20
<211>19
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>20
ggttgttgtt?gttgcagca 19
<210>21
<211>22
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>21
gctgtgcttg?gatgatactc?ta 22
<210>22
<211>19
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>22
gtggctgttg?tgaaaacga 19
<210>23
<211>19
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>23
tggggttggg?agacacata 19
<210>24
<211>19
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>24
ggcacagacg?aggaaggtt 19
<210>25
<211>19
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>25
gctgctgctg?ctgtgtaaa 19
<210>26
<211>21
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>26
aaacaacggt gatccaacta?a 21
Claims (6)
1. differentiate that wheat to be measured is the primer special that carries the wheat breed of which kind of Glu-A3 protein protomer, is made up of following seven pairs of primers:
(1) primer of the composition of Nucleotide shown in the sequence 20 of Nucleotide and sequence table shown in the sequence 18 of sequence table is right;
(2) primer of the composition of Nucleotide shown in the sequence 21 of Nucleotide and sequence table shown in the sequence 19 of sequence table is right;
(3) primer of the composition of Nucleotide shown in the sequence 22 of Nucleotide and sequence table shown in the sequence 18 of sequence table is right;
(4) primer of the composition of Nucleotide shown in the sequence 23 of Nucleotide and sequence table shown in the sequence 19 of sequence table is right;
(5) primer of the composition of Nucleotide shown in the sequence 24 of Nucleotide and sequence table shown in the sequence 18 of sequence table is right;
(6) primer of the composition of Nucleotide shown in the sequence 25 of Nucleotide and sequence table shown in the sequence 18 of sequence table is right;
(7) primer of the composition of Nucleotide shown in the sequence 26 of Nucleotide and sequence table shown in the sequence 18 of sequence table is right.
2. differentiate that wheat to be measured is the test kit that carries the wheat breed of which kind of Glu-A3 protein protomer for one kind, comprise the described primer special of claim 1.
3. the said primer special of claim 1 differentiates that in preparation wheat to be measured is the application of carrying in the test kit of wheat breed of which kind of Glu-A3 protein protomer.
4. differentiate that wheat to be measured is the method for carrying the wheat breed of which kind of Glu-A3 protein protomer for one kind, comprise the steps:
1) genomic dna with wheat to be measured is a template, carries out pcr amplification with the said primer special of claim 1;
2) detect pcr amplification product, judge that through the size of pcr amplification product wheat to be measured is the wheat breed that carries which kind of Glu-A3 protein protomer.
5. the said primer special of claim 1, the said test kit of claim 2 are differentiating that wheat is the application of carrying in the wheat breed of which kind of Glu-A3 protein protomer.
6. application as claimed in claim 5 is characterized in that: said wheat is a common wheat.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100861334A CN101570791B (en) | 2009-06-02 | 2009-06-02 | Special primer for identifying which variety of Glu-A3 protein subunit is carried in wheat as well as application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100861334A CN101570791B (en) | 2009-06-02 | 2009-06-02 | Special primer for identifying which variety of Glu-A3 protein subunit is carried in wheat as well as application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101570791A CN101570791A (en) | 2009-11-04 |
| CN101570791B true CN101570791B (en) | 2012-03-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100861334A Expired - Fee Related CN101570791B (en) | 2009-06-02 | 2009-06-02 | Special primer for identifying which variety of Glu-A3 protein subunit is carried in wheat as well as application thereof |
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| Country | Link |
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| CN (1) | CN101570791B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104480198A (en) * | 2014-11-20 | 2015-04-01 | 首都师范大学 | Molecular marker and application of low-molecular-weight glutelin Glu-A3a gene of common wheat |
| CN105734148B (en) * | 2016-04-01 | 2019-06-21 | 中国科学院遗传与发育生物学研究所 | A kind of pair of common wheat Glu-A1 gene locus difference allelic variation carries out the methods and applications of parting |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1778966A (en) * | 2004-11-18 | 2006-05-31 | 中国农业科学院作物育种栽培研究所 | Inspection of wheat polyphenol oxidase activity characteristic and its special primer |
-
2009
- 2009-06-02 CN CN2009100861334A patent/CN101570791B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1778966A (en) * | 2004-11-18 | 2006-05-31 | 中国农业科学院作物育种栽培研究所 | Inspection of wheat polyphenol oxidase activity characteristic and its special primer |
Non-Patent Citations (2)
| Title |
|---|
| Ikeda TM等.Characterization of low-molecular-weight glutenin subunit genes and their protein products in common wheats.《Theor Appl Genet》.2005,第112卷327-334. * |
| Zhang W等.Characterisation and marker development for low molecular weight glutenin genes from Glu-A3 alleles of bread wheat (Triticum aestivum.L).《Theor Appl Genet》.2004,第108卷(第7期),1409-1419. * |
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| CN101570791A (en) | 2009-11-04 |
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