CN1904050A - Wheat ear germinating resistance related gene and its application - Google Patents
Wheat ear germinating resistance related gene and its application Download PDFInfo
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Abstract
The present invention discloses a wheat head germination resistance related gene and its application. Said wheat head germination resistance related gene has one of the following nucleotide sequences: (1), nucleotide sequence of SEQ ID No.2 in sequence table; (2), DNA sequence, said DNA sequence and DNA sequence of SEQ ID No.2 in sequence table have above 90% of homology and can be used for coding protein with identical function; and (3), nucleotide sequence which can be hybridized with DNA sequence limited by SEQ ID No.2 in sequence table under the high stringent condition. Said gene can be used for screening head germination resistance wheat or head germination sensitive wheat.
Description
Technical field
The present invention relates to a kind of wheat ear germinating resistance related gene and application thereof.
Background technology
Wheat ear germinating (Pre-harvest Sprouting, be called for short PHS) be meant that wheat runs into wet weather or (the Groos C of the head sprouting under wet environment before results, Gay G, Perretant M R, Gervais L, Bernard M, DedryverF, Charmet G.Study of the relationship between pre-harvest sprouting and graincolor by quantitative trait loci analysis in a white * red grain bread-wheat cross.Theor Appl Genet, 2000,104:39-47).Because a series of biochemical reaction has taken place in its seed inside during wheat ear germinating, particularly enzymic activitys such as some carbohydrate degrading enzymes, proteolytic ferment raise, the reserve substance in degraded embryo and the endosperm, the output and the processing quality of reduction wheat.In the more time of rainfall, even the wheat breed seed outward appearance that has is not germinateed, but the nutritive substance that stores begun to transform, can influence yield and quality equally.
States such as Japan, Britain, the U.S., Canada, Australia, Brazil, Germany, Sweden, India all are subjected to fringe germination harm, and Canada and Australia are particularly serious.According to Derera, from 11 planting times of 1973-1984, Australia has 7% wheat to be subjected to the fringe harm of germinateing every year on average, and the most serious 1984, fringe germination hazard rating was up to 20%.The most of white wheat kind of northern China is germinateed extremely sensitive to fringe, meet rain harvesting time and very easily cause wheat ear germinating, and for example area, Beijing-Tianjin Ji 1995 and calendar year 2001 fringe germination are serious, cause enormous economic loss.Therefore, breeding scholar and biochemist take much count of wheat ear germinating both at home and abroad, and have carried out big quantity research at aspects such as its authentication method, resistance mechanism, hereditary mechanism and molecule markers.
The molecule marker of wheat ear germinating is domestic and international one of priority fields of study in recent years.Anderson etc. (1993) have oriented 10 RFLP marks relevant with ear germinating resistance, wherein Xcn1.bcd1434 and Xcn1.cdo431 are on 1AS karyomit(e), Xcn1.cdo64 is on 2AS karyomit(e), Xcn1.Wg996a and Xcn1.bcd120a are on 2AL karyomit(e), Xcn1.bdcd450 is on 5DL karyomit(e), Xcn1.cdo545 is on 4AL karyomit(e), Xcn1.bcd450 is on 5DL karyomit(e), Xcn1.bcd426 is (Anderson J A on 6BL karyomit(e), Sorrells M E, Tanksley SD.RFLPanalysis of genomic regions associated with resistance to pre-harvest sproutingin wheat.Crop Sci, 1993,33:453-459).
Roy etc. (1999) utilize STMS (Sequence-tagged Microsatellites) to be marked at and have located 1 major gene that the control fringe germinates on the 6BS karyomit(e), with Cwmc104 mark close linkage.Utilize STS (Sequence-tagged site) to be marked at and also located 1 major gene that the control fringe germinates on the 7DL karyomit(e), with Xmst101 mark close linkage (Roy J K, Prasad M, Varshney R K.Identification of amicrosatellite on chromosomes 6B and a STS on 7D of bread wheat showing anassociation with pre-harvest sprouting tolerance.Theor Appl Genet, 1999,99:336-340).Zanetti etc. (2000) have also located QTL (ZanettiS on 2A, 3B, 5A, 6A and 7B karyomit(e), Winzeler M, Keller M, Keller B, MessmerM.Genetic analysis of pre-harvestsproutingin a wheat * spelt cross.Crop Sci, 2000,40:1406-1407).
Flintham etc. (2000) study the dependency of dormancy and R gene with the near isogenic line of one group of red grain and white grain.Result of study shows that the R gene is a kind of triple gene of controlling red grain look, is positioned at the chromosomal long arm end of third part homology group, that is to say in A, B and the D genome and all contains the R gene locus, and the color of its control kind of skin belongs to matrocliny.The R gene of different sources has the double effect of dormancy and seed color (Flintham JE (2000) Differentgenetic components control coat-imposed and embryo-imposed dormancy in wheat.SeedSci Res 10:43-50).Flintham is interpreted as this species diversity also existing in embryo one with dormancy the major gene Phs of substantial connection to be arranged, and this gene was located on the 4AL afterwards, was gene locus (the Flintham JE that influences dormancy and after-ripening level, Adlam R, Bassoi M, Holdsworth M ， ﹠amp; Gale M.Mapping genes forresistance to sprouting damage inwheat.Euphytica, 2002,126:39-45).Himi (2005) has proved that the R gene can promote the flavonoid gene transcription, in white grain kind, owing to having to express, the R gene do not make the flavonoid expression of gene reduce (Himi E, Noda K.Red grain color gene (R) of wheat is a Myb-typetranscription factor.Euphytica, 2005,143:239-242).R gene locus and dormant gene close linkage are so in the wheat ear germinating resistance breed breeding, the kernel seed coat colour that the R gene is controlled can be used as the mark of an ear germinating resistance and uses.But this mark can not be applied to the marker-assisted breeding of white wheat ear germinating resistance.
In addition, also have the QTL of many ear germinating resistances to be positioned on the different chromosome segments, comprise awns property (the Derera N F.Preharvest field sproutingin cereal.CRC Press that Hd, the BI on the 5AL on the 4BS and the sites such as B2 on the 6BL are controlled, Inc, 1989), the fringe type of the C site control on the 2DL, site (the King R W.﹠amp of the control upper epidermis wax of finding on 2B and the 2D site; P.von Wettstein-Knowles.Epicuticular waxesand regulation of ear wetting and pre-harvest sproutlng in barley and wheat.Euphytica, 2000,112:157-166), and the dwarf gene site of on 4B and 4D galianconism, finding, all with relevant (the Flintham JE of reduction amylase activity, Angus WJ, Gale MD, Heterosis, overdominance forgrain yield, and alpha-anylase activity in F1 hybirds between nearisogenic Rhtdwarf and tall wheat.Journal of Agricultural Science, 1997,129:371-378).Though the mark of awns property, fringe type, plant height and seed epicutile wax is relevant with ear germinating resistance, they are not the principal element that influences ear germinating resistance, so these marks can be used seldom in breeding.
In sum, although the molecule marker that the wheat ear germinating resistance of having developed is correlated with is a lot, great majority are AFLP and SSR mark, poor practicability, and await further checking.
Nakamura (2001) has studied Vp-1 expression of gene among strong dormancy kind Minamino and the non-dormancy kind Tozan, the expression amount that found that the Vp-1 of Minamino in mature embryo is greater than Tozan (Nakamura S, Toyama T (2001) Isolation of a VP1 homologue from wheat and analysis of its expression inembryos of dormant and non-dormant cultivars.J Exp Bot 52:875-876).The sequence of Vp-1B gene is seen sequence 1, and the coded protein sequence of this sequence is seen sequence 4.
Summary of the invention
An object of the present invention is to provide a kind of wheat ear germinating resistance related gene.
Wheat ear germinating resistance related gene provided by the present invention, name is called Vp-1Bb, has one of following nucleotide sequence:
1) SEQ ID № in the sequence table: 2 nucleotide sequence;
2) with sequence table in SEQ ID №: 2 dna sequence dna has 90% above homology, and the identical function protein DNA sequence of encoding;
3) under the rigorous condition of height can with the SEQ ID № in the sequence table: the nucleotide sequence of the 2 dna sequence dnas hybridization that limit.
The rigorous condition of above-mentioned height can be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, under 65 ℃, hybridize and wash film.
The Vp-1Bb gene is the allelic variation of Vp-1B gene, has inserted the nucleotide sequence of 193bp in the 3rd intron of Vp-1B gene.
The Vp-1B gene has the nucleotide sequence of sequence 1 in the sequence table, total length 4034bp.The Vp-1B gene is made up of 6 exons and 5 introns, from 5 of sequence 1 ' the 1st to 1527 deoxynucleotides of end is first exon, from 5 of sequence 1 ' the 1528th to 1737 deoxynucleotides of end is first intron, from 5 of sequence 1 ' the 1738th to 1858 deoxynucleotides of end is second exon, from 5 of sequence 1 ' the 1859th to 2181 deoxynucleotides of end is second intron, from 5 of sequence 1 ' the 2182nd to 2282 deoxynucleotides of end is the 3rd exon, from 5 of sequence 1 ' the 2283rd to 2903 deoxynucleotides of end is the 3rd intron, from 5 of sequence 1 ' the 2904th to 2950 deoxynucleotides of end is the 4th exon, from 5 of sequence 1 ' the 2951st to 3051 deoxynucleotides of end is the 4th intron, from 5 of sequence 1 ' the 3052nd to 3128 deoxynucleotides of end is the 5th exon, from 5 of sequence 1 ' the 3129th to 3644 deoxynucleotides of end is the 5th intron, is the 6th exon from 5 of sequence 1 ' the 3645th to 4034 deoxynucleotides of end.The Vp-1B genes encoding has the aminoacid sequence of sequence 4 in the sequence table.The Vp-1Bb gene has the nucleotide sequence of sequence 2 in the sequence table.The Vp-1Bb gene has inserted 193bp the Vp-1B gene between 5 of sequence 2 ' the 2497th to 2498 deoxynucleotides of end.The Vp-1Bb gene is made up of 6 exons and 5 introns, from 5 of sequence 2 ' the 1st to 1527 deoxynucleotides of end is first exon, from 5 of sequence 2 ' the 1528th to 1737 deoxynucleotides of end is first intron, from 5 of sequence 2 ' the 1738th to 1858 deoxynucleotides of end is second exon, from 5 of sequence 2 ' the 1859th to 2181 deoxynucleotides of end is second intron, from 5 of sequence 2 ' the 2182nd to 2282 deoxynucleotides of end is the 3rd exon, from 5 of sequence 2 ' the 2283rd to 3096 deoxynucleotides of end is the 3rd intron, from 5 of sequence 2 ' the 3097th to 3143 deoxynucleotides of end is the 4th exon, from 5 of sequence 2 ' the 3144th to 3245 deoxynucleotides of end is the 4th intron, from 5 of sequence 2 ' the 3246th to 3322 deoxynucleotides of end is the 5th exon, from 5 of sequence 2 ' the 3323rd to 3838 deoxynucleotides of end is the 5th intron, is the 6th exon from 5 of sequence 2 ' the 3839th to 4227 deoxynucleotides of end.The Vp-1Bb genes encoding has the protein of the aminoacid sequence of sequence 4 in the sequence table.
The recombinant expression vector, transgenic cell line and the engineering bacteria that contain Vp-1Bb gene of the present invention all belong to protection scope of the present invention.
Another object of the present invention provides a kind of assisting sifting ear germinating resistance method of wheat.
Assisting sifting ear germinating resistance method of wheat provided by the present invention, be to detect the nucleotide sequence that whether has inserted 193bp in the 3rd intron of Vp-1B gene of wheat to be measured, inserted the nucleotide sequence of 193bp in the 3rd intron as the Vp-1B gene of this wheat to be measured, then this wheat to be measured is candidate's ear germinating resistance wheat.
Available PCR method detects the nucleotide sequence that whether has inserted 193bp in the 3rd intron of Vp-1B gene of wheat to be measured.
Described PCR method is a template for the genomic dna with wheat to be measured, use a pair of primer of forming by the nucleotide sequence of sequence in the sequence table 5 and sequence 6 to carry out pcr amplification, if there is size to be that the band of 845bp, then described wheat to be measured are candidate's ear germinating resistance wheat breed in the pcr amplification product that obtains.
The a pair of primer name of being made up of the nucleotide sequence of sequence in the sequence table 5 and sequence 6 is called Vp1B3, be according to the ear germinating resistance relevant STS mark of Vp-1 gene, be positioned at 3BL in the design of the difference between ear germinating resistance on the B genome and the sensitive material.
The method of described detection amplified production is amplified production to be carried out 2.0% agarose detection, 130V voltage electrophoresis 1.5 hours.
The 3rd purpose of the present invention provides a kind of method of assisting sifting fringe germination sensitive wheat.
The method of assisting sifting fringe germination sensitive wheat provided by the present invention, be to detect to have or not in the 3rd intron of Vp-1B gene of wheat to be measured to insert and deletion mutantion, do not insert and deletion mutantion as having in the 3rd intron of the Vp-1B gene of this wheat to be measured, then this wheat to be measured is candidate's fringe germination sensitive wheat; Described insertion sports the sudden change of inserting the nucleotide sequence of 193bp in the 3rd intron of Vp-1B gene, and described deletion mutantion is the sudden change of the nucleotide sequence of disappearance 83bp in the 3rd intron of Vp-1B gene.
It is to have or not to insert and deletion mutantion that available PCR method detects in the 3rd intron of Vp-1B gene of wheat to be measured.
Described PCR method is a template for the genomic dna with wheat to be measured, use a pair of primer of forming by the nucleotide sequence of sequence in the sequence table 5 and sequence 6 to carry out pcr amplification, if there be not the band of size for 845bp and 569bp in the pcr amplification product that obtains, then described wheat to be measured is candidate's fringe germination sensitive wheat.
The method of assisting sifting ear germinating resistance of the present invention or sensitive wheat is particularly suitable for the white wheat kind.
The method of assisting sifting ear germinating resistance of the present invention or sensitive wheat can be used for the morning of wheat ear germinating resistance breeding for selection, shortened breeding cycle greatly, accelerate breeding speed, reduce the breeding cost, have simple to operate, with low cost, the advantage that cycle is short, be suitable for applying, for the breeding of wheat anti growing out provides technical support, for seed selection ear germinating resistance wheat germplasm provides a kind of system of selection efficiently.
Description of drawings
The banding pattern that Fig. 1 increases in 17 different ear germinating resistance kinds for Vp1B3
Fig. 2 is 84 different ear germinating resistance materials and three kinds of segmental histograms that concern with Vp1B3 amplified
Embodiment
Wheat lines is all available from national germ plasm resource gene pool among the following embodiment.
The present invention has utilized common wheat Vp-1 gene conservative sequences Design primer increases to A, B and the genomic Vp-1 gene of D respectively, does not find differences on A genome and D genome; And on the B genome, compare with fringe germination sensitive material, the ear germinating resistance material exists the insertion of a 193bp or the disappearance of 83bp in the 3rd intron, insert the type major part and be present in the high and highly stable several local variety of the ear germinating resistance of minority, the disappearance type is present in the resistant variety.Designed with an ear germinating resistance relevant STS mark-Vp1B3 at ear germinating resistance on the B genome with difference between the sensitive material according to the Vp-1 gene, the result shows that this mark presents polymorphism, the fragment of 652bp appears in the fragment that occurs 845bp or 569bp in resistant variety in perceptual kind.Further two allelic variation VP-1Bb and the VP-1Bc that the ear germinating resistance kind is increased and obtains the Vp-1B gene.
Carry out the ear germinating resistance evaluation with seed germination index GI as the index of ear germinating resistance, seed germination index GI is the percentage ratio that total number of shoots of the 7th day accounts for total seed number.Seed germination index GI is a fringe germination sensitive varieties greater than 70%, and seed germination index GI is that 30-50% is the fringe neutral kind of germinateing, and germination index GI is that 0-15% is the ear germinating resistance kind.
The allelic variation VP-1Bb of embodiment 1, two Vp-1B genes and VP-1Bc acquisition
1, the allelic variation VP-1Bb of two Vp-1B genes and the discovery of VP-1Bc
With the white wheat in high anti-material Wan County (germination index GI is 8%) and ear germinating resistance material west farming 979 (germination index GI is 15%) of germinateing of excellent 9507 (germination index GI is 89%) and fringe in the fringe germination sensitive material is material, its Vp-1 gene is carried out the segmentation amplification at A, B and D genome, order-checking then, the result shows that the Vp-1 gene does not find differences on A genome and D genome; And on the B genome, with excellent 9507 compare in the fringe germination sensitive material, in the 3rd intron, there are differences the insertion that promptly in fringe germinates the white wheat in high anti-material Wan County, has a 193bp in the ear germinating resistance material, in ear germinating resistance material west farming 979, there is the disappearance of a 83bp.The Vp-1B allelotrope called after Vp-1Bb of 193bp will be in the 3rd intron of the genomic Vp-1 gene of B (being the Vp-1B gene), inserted, the Vp-1B allelotrope called after vp-1Bc of 83bp will be in the 3rd intron of the genomic Vp-1 gene of B (being the Vp-1B gene), lacked.The Vp-1B gene has the nucleotide sequence of sequence 1 in the sequence table.The Vp-1B gene is made up of 6 exons and 5 introns, from 5 of sequence 1 ' the 1st to 1527 deoxynucleotides of end is first exon, from 5 of sequence 1 ' the 1528th to 1737 deoxynucleotides of end is first intron, from 5 of sequence 1 ' the 1738th to 1858 deoxynucleotides of end is second exon, from 5 of sequence 1 ' the 1859th to 2181 deoxynucleotides of end is second intron, from 5 of sequence 1 ' the 2182nd to 2282 deoxynucleotides of end is the 3rd exon, from 5 of sequence 1 ' the 2283rd to 2903 deoxynucleotides of end is the 3rd intron, from 5 of sequence 1 ' the 2904th to 2950 deoxynucleotides of end is the 4th exon, from 5 of sequence 1 ' the 2951st to 3051 deoxynucleotides of end is the 4th intron, from 5 of sequence 1 ' the 3052nd to 3128 deoxynucleotides of end is the 5th exon, from 5 of sequence 1 ' the 3129th to 3644 deoxynucleotides of end is the 5th intron, is the 6th exon from 5 of sequence 1 ' the 3645th to 4034 deoxynucleotides of end.The Vp-1Bb gene has the nucleotide sequence of sequence 2 in the sequence table, and the Vp-1Bb gene has inserted 193bp the Vp-1B gene between 5 of sequence 1 ' the 2497th to 2498 deoxynucleotides of end.The Vp-1Bb gene is made up of 6 exons and 5 introns, from 5 of sequence 2 ' the 1528th to 1737 deoxynucleotides of end is first intron, from 5 of sequence 2 ' the 1738th to 1858 deoxynucleotides of end is second exon, from 5 of sequence 2 ' the 1859th to 2181 deoxynucleotides of end is second intron, from 5 of sequence 2 ' the 2182nd to 2282 deoxynucleotides of end is the 3rd exon, from 5 of sequence 2 ' the 2283rd to 3096 deoxynucleotides of end is the 3rd intron, from 5 of sequence 2 ' the 3097th to 3143 deoxynucleotides of end is the 4th exon, from 5 of sequence 2 ' the 3144th to 3245 deoxynucleotides of end is the 4th intron, from 5 of sequence 2 ' the 3246th to 3322 deoxynucleotides of end is the 5th exon, from 5 of sequence 2 ' the 3323rd to 3838 deoxynucleotides of end is the 5th intron, is the 6th exon from 5 of sequence 2 ' the 3839th to 4227 deoxynucleotides of end.The Vp-1Bc gene has the nucleotide sequence of sequence 3 in the sequence table, and the Vp-1Bc gene has lacked 83bp the vp-1B gene between 5 of sequence 1 ' the 2710th to 2794 deoxynucleotides of end.The Vp-1Bc gene is made up of 6 exons and 5 introns, from 5 of sequence 3 ' the 1st to 1527 deoxynucleotides of end is first exon, from 5 of sequence 3 ' the 1528th to 1737 deoxynucleotides of end is first intron, from 5 of sequence 3 ' the 1738th to 1858 deoxynucleotides of end is second exon, from 5 of sequence 3 ' the 1859th to 2181 deoxynucleotides of end is second intron, from 5 of sequence 3 ' the 2182nd to 2282 deoxynucleotides of end is the 3rd exon, from 5 of sequence 3 ' the 2283rd to 2820 deoxynucleotides of end is the 3rd intron, from 5 of sequence 3 ' the 2821st to 2867 deoxynucleotides of end is the 4th exon, from 5 of sequence 3 ' the 2868th to 2968 deoxynucleotides of end is the 4th intron, from 5 of sequence 3 ' the 2969th to 3045 deoxynucleotides of end is the 5th exon, from 5 of sequence 3 ' the 3046th to 3561 deoxynucleotides of end is the 5th intron, is the 6th exon from 5 of sequence 3 ' the 3562nd to 3951 deoxynucleotides of end.
2, the acquisition of Vp-1B gene, VP-1Bb gene and VP-1Bc gene
With in excellent 9507 genomic dna be template, with Vp-1B
3F/R (5 '-TGCTCCTTTCCCAATTGG-3 '; 5 '-ACCCTCCTGCAGCTCATT-3 '), obtains having the Vp-1B gene of the nucleotide sequence of sequence 1 in the sequence table for primer carries out pcr amplification.Wherein, the pcr amplification condition is 94 ℃ of pre-sex change of elder generation 5 minutes; Carry out 36 circulations then, 94 ℃ of sex change of each circulation 1 minute, 60 ℃ of renaturation 1 minute, 72 ℃ were extended 1 minute; Last 72 ℃ were extended 10 minutes.
Genomic dna with the white wheat in Wan County is a template, with Vp-1B
3F/R (5 '-TGCTCCTTTCCCAATTGG-3 '; 5 '-ACCCTCCTGCAGCTCATT-3 '), obtains having the Vp-1Bb gene of the nucleotide sequence of sequence 2 in the sequence table for primer carries out pcr amplification.Wherein, the pcr amplification condition is 94 ℃ of pre-sex change of elder generation 5 minutes; Carry out 36 circulations then, 94 ℃ of sex change of each circulation 1 minute, 60 ℃ of renaturation 1 minute, 72 ℃ were extended 1 minute; Last 72 ℃ were extended 10 minutes.
To the west of farming 979 genomic dna be template, with Vp-1B
3F/R (5 '-TGCTCCTTTCCCAATTGG-3 '; 5 '-ACCCTCCTGCAGCTCATT-3 '), obtains having the Vp-1Bc gene of the nucleotide sequence of sequence 3 in the sequence table for primer carries out pcr amplification.Wherein, the pcr amplification condition is 94 ℃ of pre-sex change of elder generation 5 minutes; Carry out 36 circulations then, 94 ℃ of sex change of each circulation 1 minute, 60 ℃ of renaturation 1 minute, 72 ℃ were extended 1 minute; Last 72 ℃ were extended 10 minutes.
Difference assisting sifting ear germinating resistance wheat and fringe germination sensitive wheat between the 3rd intron of embodiment 2, usefulness Vp-1B, VP-1Bb and VP-1Bc gene
1, according to the sequences Design of Vp-1B, VP-1Bb and VP-1Bc STS mark Vp1B3, primer sequence is: the upstream: 5 '-TGCTCCTTTCCCAATTGG-3 (sequence 5); Downstream: 5 '-ACCCTCCTGCAGCTCATT G-3 ' (sequence 6).Use the white grain kind (Fig. 1) of 17 different ear germinating resistances of this primer amplification then, amplified reaction is an example with 20 μ l totally, and reaction solution consists of 2 μ l, 10 * PCR buffer, 4 kinds of each 125 μ M of dNTP, the upstream and downstream primer of each 8pmol, the rTaq enzyme of 1.0 units and 50ng wheat cdna group DNA.Response procedures is 94 ℃ of pre-sex change of elder generation 5 minutes; Carry out 36 circulations then, 94 ℃ of sex change of each circulation 1 minute, 60 ℃ of renaturation 1 minute, 72 ℃ were extended 1 minute; Last 72 ℃ were extended 10 minutes.The PCR product is stored in 4 ℃.Use 2.0% sepharose at 1.5 hours electrophoresis detection PCR of 130V voltage electrophoresis product then.The result three kinds of banding patterns occur: the fragment of 845bp or 569bp occurs in resistant variety, the fragment of 652bp occurs in perceptual kind as shown in Figure 1.Dna molecular amount standard among Fig. 1 is 100bp DNAladder.Among Fig. 1,1 is that (germination index GI is 91.5% to CA9722, the 652bp band increases), 2 for rich anti-13 (germination index GI is 98.5%, the 652bp band increases), 3 is that (germination index GI is 4.0% in interior township 173, the 569bp band increases)), 4 is that No. 9 (germination index GI is 11% in the mountain farming, the 569bp band increases), 5 is that No. 5 (germination index GI is 97.5% to the Shanxi wheat, the 652bp band increases), 6 is that (germination index GI is 0.5% in another name for Sichuan Province 10,000, the 569bp band increases), 7 is that (germination index GI is 88.5% in Shijiazhuang 54, the 652bp band increases), 8 is that (germination index GI is 2.5% to SW95-16117, the 569bp band increases), 9 is that (germination index GI is 5.5% to river farming 94-DH343, the 569bp band increases), 10 are white jade skin (germination index GI is 7.5%, and the 569bp band increases), 11 is that (germination index GI is 52.0% in Changle 5, the 652bp band increases), 12 are Huixian red (germination index GI is 15.0%, and the 652bp band increases), 13 is that (germination index GI is 6.0% in Shaanxi 54, the 569bp band increases), 14 are section's something lost 26 (germination index GI is 96.5%, and the 652bp band increases), 15 for the river educates 21729 (germination index GI is 7.5%, the 845bp band increases), 16 are Nanjing University 2419 (germination index GI is 4.5%, and the 569bp band increases), 17 are foreign wheat (germination index GI is 1.0%, and the 845bp band increases) morning.
2, the white grain kind of choosing 84 different ear germinating resistances detects the feasibility of assisting sifting ear germinating resistance of the present invention or sensitive wheat.
Genomic dna with the white grain kind of 84 different ear germinating resistances is a template respectively, method according to step 1 increases and electrophoresis, the result shows: have the fragment that 8 kinds amplify 845bp, wherein 3 kinds are ear germinating resistance material (germination index GI are 0-15%), and other 5 kinds are anti-materials (germination index GI is 31%-50%) during fringe germinates; 39 kinds amplify the fragment of 569bp, and wherein 30 kinds are ear germinating resistance material (germination index GI are 0-15%), 7 kinds be in anti-material, other 2 kinds are the fringe perceptual materials that germinate.37 kinds amplify the fragment of 652bp, and 20 kinds are fringe germination sensitive materials, and 15 kinds are anti-materials during fringe germinates, and other 2 materials are ear germinating resistance materials.And the material with 845bp is than the anti growing out more of the material with 569bp.
84 different ear germinating resistance materials with use Vp1B
3Three kinds of segmental relations that amplified as shown in Figure 2.Among Fig. 2, the Bai Zhu segmental kind number of 569bp of representing to increase, black post is to amplify the segmental kind number of 652bp, gray columns is to amplify the segmental kind number of 845bp.
The fringe germination index GI of the table 1.84 part Chinese wheat breed of white grain and the polymorphism of STS mark
| Number | Variety name | Fringe germination index GI | PCR fragment (bp) | Number | Variety name | Fringe germination index GI | PCR fragment (bp) |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 | 15 another name for Sichuan Province 10,000, Mianyang early in the foreign wheat river 81-11108 SW 95-16117 township's 173 peaces select the big 2419 little 5 Mianyang 8186-0-14 that lay down white shell in 2 south early to educate little No. 1, No. 9 white baldhead Bi Ma of Da Kou wheat mountain, No. 1 Luoyang, 168 Shandong wheats, 21 orchid wheat Xinyang farming of laying down of 21729 Mianyang 94-315 west farming 6028 in 54 mountains, 19 Shaanxi, 5762 interior township, farming 94-DH343 Xichang, 2 rivers farming, 6521 Plains, 50 Mianyang 93-352 neat big 195 rivers of 124 white jade skins of breaking a seal | 0.0 0.5 1.0 2.0 2.5 4.0 4.0 4.5 4.5 5.0 5.0 5.5 5.5 6.0 6.0 6.0 6.0 6.5 7.0 7.5 7.5 7.5 7.5 8.0 8.0 8.5 8.5 9.0 11 11.0 12.5 12.5 | 569 569 845 569 569 569 569 569 569 569 569 569 569 569 569 569 569 569 569 569 569 845 569 569 569 569 845 569 569 569 569 569 | 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 | No. 2 Jimais of wheat are spent among 54 rich anti-No. 4 Jimai 21 CA9722 of No. 1 Shijiazhuang No. 1 in No. 5 Henan wheats in No. 5 former rich No. 1 Henan wheat 50 Henan wheats, 15 Changle, No. 3 Mount Taishan, No. 5 Zhengzhou of 928802 Henan wheats, 62 bar wheats, No. 4 951741 Shandong, 407 flat fringe wheat Jinan, agricultural 88 PH, 8516 Shijiazhuang, No. 1 west of 7023 Shandong wheats, No. 2 Linfen of Shandong wheat 21 little No. 10 capital of 411 former winters 3 Shanxi wheats, 22 935031 all | 38.0 39.0 39.5 39.5 42.0 43.5 43.5 44.0 46.0 46.5 47.0 49.0 49.5 50.0 50.0 50.0 50.5 52.0 52.0 54.0 70.5 78.5 81.5 83.5 84.0 87.0 84.5 89.5 90.0 91.5 92.5 95.0 | 845 652 569 652 652 652 845 845 652 652 652 569 652 845 652 569 652 652 652 652 652 652 652 652 652 569 652 652 652 652 569 652 |
| 33 34 35 36 37 38 39 40 41 42 | No. 3 rich anti-No. 6 Jingyangs 302, No. 3 Shandong wheat No. 7 Zhengzhou of 12 Henan wheats, 34 Shandong wheats of No. 8 mountain farmings of red Henan, the white Huixian of red fringe wheat | 13.5 15.0 31.0 32.0 33.0 33.5 34.0 34.0 36.0 38.0 | 652 569 569 845 569 652 569 652 652 569 | 75 76 77 78 79 80 81 82 83 84 | Capital 437 morals select No. 1 section to lose No. 9 rich anti-13 Ji | 95.0 34.5 96.5 35.5 96.5 97.5 97.5 98.0 98.5 99.0 | 652 652 652 652 652 652 652 652 652 652 |
Sequence table
<160>6
<210>1
<211>4034
<212>DNA
＜213〉excellent 9507 (Triticum aestivum L cv Zhongyou 9507) in the Triticum common wheat
<400>1
atggacgcct ccgccggctc gtcgccgccg cggcactcgc agggggacgc gccgaggcgc 60
ggcgggccgc accgcgggaa gggccccgcg gtggagatcc ggcagggaga ggacgacttc 120
atgttcgcgc aggatacctt cccggccctc ccggacttcc cttgcctctc ctcgccgtcg 180
agctccacct tctcctcctc ctcgtcttcc aactcctcca gcgccttcgc tgccccagcg 240
ggagcgggcg ggcgcggggc cgagggggcg cgcggcgagc cgtccgagcc tgcagcggcc 300
ggggacggga tggacgacct ctccgacatc gaccacctgc tcgacttcgc gtccatcaac 360
gacgacgtcc cctgggacga cgagccgctc ttccccgacg tcgggatgat gctggaggat 420
gtcatctccg agcagcagct ccagcagcct ccggcgggcc acggcacggc cgggagaacg 480
gcgtcgcatg cggcctctgg tggaggagag gatgccttca tgggtggcgg cggcacgggg 540
agcgcggcgg acgacctgcc gctgttcttc atggagtggc tcaagaacaa ccgcgactgc 600
atctcggccg aggacctccg cagcatccgc ctccgtcgat ccaccatcga ggccgcggcc 660
gcgcgcctcg gtggggggcg ccagggcacc atgcagctgc tcaagctcat cctcacctgg 720
gtgcagaacc accacctgca gaagaagcgc ccccgcgtcg gcgccatgga tcaggaggcg 780
ccgccggcag gaggccagct ccctagcccc ggcgcaaacc ccggctacga attccccgcg 840
gagacgggtg ccgccgctgc cacatcatgg atgccctacc aggccttctc gccaactgga 900
tcctacggcg gcgaggcgat ctacccattc cagcagggct gcagcactag cagcgtggcc 960
gtgagcagcc agccgttctc cccgccggcg gcgcccgaca tgcacgccgg ggcctggccc 1020
ctgcagtacg cggcgttcgt cccagctgga gccacatccg caggcactca aacatacccg 1080
atgccgccgc cgggggccgt gccgcagccg ttcgcggccc ccggattcgc cgggcagttc 1140
ccgcagcgga tggagccggc ggcgaccagg gaggcccgga agaagcggat ggcgaggcag 1200
cggcgcttgt cgtgcctgca gcagcagcgg agccagcagc tgaatctgag ccagatccaa 1260
accggcggct tccctcaaga gcaatccccc cgcgcggcgc actcggcgcc ggtcacgccg 1320
ccgtcgtccg gctggggagg cctctggacg caacaggccg tccagggcca gctcatggtc 1380
caggtcccga atccgctgtc gacgaagtcc aattcctcaa ggcagaagca gcaaaaaccc 1440
tcgccggacg cagcagcgag gccgccctcc ggcggcgccg ccacgccgca tcgcccgggg 1500
caggcgtcgg cttccaataa gcagcggcag caggtgcatg catgcacgaa cacctcttgc 1560
catccatcca tcgatcgcca tcccgcatag aatcacaagc cattgctccc caaataaagt 1620
gtgcgtgctt cgtaatggac gcacatcgct gtccagcgat aggatatgca tgcgcgcgtg 1680
cgtgtctgtt ttacctgcac ggtgaattgc gtttctcaac gaggttccgt gcatgcgcgc 1740
agggtgcgag gacgccggcg gcggcgccgg cggcaggaga caagaacctg cggttcctgc 1800
tgcagaaggt gctcaagcag agcgacgtcg gaaccctcgg ccgcatcgtg ctccccaaag 1860
tgaggcagtc acctccctat cgcggtcttc ttcctccatc cgcccgaccc atgcaccact 1920
tgcaattttc agctatatat gcatgcatcc acttgcaaat tttagtttct ccttgattcc 1980
tctttgatta gacaagatct atctttggat tgcgcctgta tgacctagct atgaatggat 2040
cttcctttct tttctgtaat ctttgtgttg atttcttctt gatttctcgg attgcaagtg 2100
tatctatcca tggatcttcc tgtctttctg ctgatcgatc ggttgcgact aaaaagtgtc 2160
tcttcttcca tggatggttt cagaaggaag cggagactca cctgccggag ctcaagacgg 2220
gggacggcat ctcgatcccc attgaggaca tcggcacatc tcaggtctgg agcatgcggt 2280
accggtaagt agtactcccg tcgatcgatt ttgggataaa tttaataata atattaaaca 2340
tgcatgcttt cattagtcca cttttatctt tttttaatta tctgtaggcc acatctagta 2400
aatgacatga tcactctgca tcactgtggg acacgcatgc tcctttccca attgggcaag 2460
tgggtcatat gaatacatat gatgggaatc cgggagcatc cgggagcaaa attgctttac 2520
tgcgctctct ctatatatac gtgaacgggc gggcatgcag aaattttgtt ttcgcatcgt 2580
ctcttggttc actccttgtt agcatcactc tagacagtat attatcataa tgaagtggtg 2640
tttctgatac catacttcct atttttccgt cttcttcttc ttcttccgaa gtgtatcata 2700
tgttgctagt tactccctcc gtaaactaat ataagagcgt ttagattact attttagtga 2760
tctaaatgct cttatattag tttacagagg gagtaattag ctagagctat caaaatgaga 2820
aaaaaagaac tagctagttt gtattctgca tgcatgataa caaattgctg gtacaatgat 2880
ggctcttgat ttttcatgca tgcagatttt ggcccaacaa caagagcaga atgtatcttc 2940
tagagaacac tggtgagaga agagaagcaa acttgcctga tacaattttg aaaccaaata 3000
tgttgctagc ttcccttgtg ttcacaaaac gtttcttctg cttgaccttg taggtgactt 3060
tgttcggtcc aatgagctgc aggagggtga tttcatcgtg ctttactctg atgtcaagtc 3120
aggcaaatat gtaatgatca tacccttccc gttaattcat ttccatgaaa tgatcggttt 3180
acatccatat gcatgttgcg aactgatcag ttgctgccat tttgggagcg ttggcatata 3240
attatcacaa tatatactat cccgtccaaa aatataatac ggattattat gtactatgag 3300
gtcaacgttt taaaactttg gcctcttgta tgtgcgtaag tatatgcatg gatattgtgc 3360
atatattaat agcaataaca tcgtattcat cttgcaaaat agttttatat catgcatatc 3420
tccctattct ttatgcatat tatgaggtca aaatcaagtg ttggagactg aaaagtcaga 3480
tgtctatgtc ttgtattctg tgagggatgg agtatatagg actgtttgtc tttggagttt 3540
ggatatctaa ttaagtacgt actagcagaa ctatctgtgt tcttgttgtg catttgaatg 3600
aattgtctat acatcagcta atctcagcct ttctgtgtac caccagctga tacgcggcgt 3660
gaaggtaaga gctcaacagg atctagccaa gcacaagaat gccagtccag agaaaggtgg 3720
ggcgtccgac gtgaaggcgg gcggagaaga cggtggttgc aaagagaagt ctccgcacgg 3780
tgtccggcga tctcgccagg aagccagctc catgaaccag atggcggtga gcatctgaaa 3840
tgagcaggct cgccgtccga tcccccattg aagactactt aattagctag ctcaagtata 3900
cctgctaatg atgatcaaat cgatctcccg ttctatgatc cgtgcttccg tgtactgccg 3960
tagccctagt tagggatgat gatactaaag tagctatcgg tttcttggtc agatgtgacg 4020
ctgaagaatg gcat 4034
<210>2
<211>4227
<212>DNA
＜213〉the white wheat in Triticum common wheat Wan County (Triticum aestivum L cv Wanxian white wheat)
<400>2
atggacgcct ccgccggctc gtcgccgccg cggcactcgc agggggacgc gccgaggcgc 60
ggcgggccgc accgcgggaa gggccccgcg gtggagatcc ggcagggaga ggacgacttc 120
atgttcgcgc aggatacctt cccggccctc ccggacttcc cttgcctctc ctcgccgtcg 180
agctccacct tctcctcctc ctcgtcttcc aactcctcca gcgccttcgc tgccccagcg 240
ggagcgggcg ggcgcggggc cgagggggcg cgcggcgagc cgtccgagcc tgcagcggcc 300
ggggacggga tggacgacct ctccgacatc gaccacctgc tcgacttcgc gtccatcaac 360
gacgacgtcc cctgggacga cgagccgctc ttccccgacg tcgggatgat gctggaggat 420
gtcatctccg agcagcagct ccagcagcct ccggcgggcc acggcacggc cgggagaacg 480
gcgtcgcatg cggcctctgg tggaggagag gatgccttca tgggtggcgg cggcacgggg 540
agcgcggcgg acgacctgcc gctgttcttc atggagtggc tcaagaacaa ccgcgactgc 600
atctcggccg aggacctccg cagcatccgc ctccgtcgat ccaccatcga ggccgcggcc 660
gcgcgcctcg gtggggggcg ccagggcacc atgcagctgc tcaagctcat cctcacctgg 720
gtgcagaacc accacctgca gaagaagcgc ccccgcgtcg gcgccatgga tcaggaggcg 780
ccgccggcag gaggccagct ccctagcccc ggcgcaaacc ccggctacga attccccgcg 840
gagacgggtg ccgccgctgc cacatcatgg atgccctacc aggccttctc gccaactgga 900
tcctacggcg gcgaggcgat ctacccattc cagcagggct gcagcactag cagcgtggcc 960
gtgagcagcc agccgttctc cccgccggcg gcgcccgaca tgcacgccgg ggcctggccc 1020
ctgcagtacg cggcgttcgt cccagctgga gccacatccg caggcactca aacatacccg 1080
atgccgccgc cgggggccgt gccgcagccg ttcgcggccc ccggattcgc cgggcagttc 1140
ccgcagcgga tggagccggc ggcgaccagg gaggcccgga agaagcggat ggcgaggcag 1200
cggcgcttgt cgtgcctgca gcagcagcgg agccagcagc tgaatctgag ccagatccaa 1260
accggcggct tccctcaaga gcaatccccc cgcgcggcgc actcggcgcc ggtcacgccg 1320
ccgtcgtccg gctggggagg cctctggacg caacaggccg tccagggcca gctcatggtc 1380
caggtcccga atccgctgtc gacgaagtcc aattcctcaa ggcagaagca gcaaaaaccc 1440
tcgccggacg cagcagcgag gccgccctcc ggcggcgccg ccacgccgca tcgcccgggg 1500
caggcgtcgg cttccaataa gcagcggcag caggtgcatg catgcacgaa cacctcttgc 1560
catccatcca tcgatcgcca tcccgcatag aatcacaagc cattgctccc caaataaagt 1620
gtgcgtgctt cgtaatggac gcacatcgct gtccagcgat aggatatgca tgcgcgcgtg 1680
cgtgtctgtt ttacctgcac ggtgaattgc gtttctcaac gaggttccgt gcatgcgcgc 1740
agggtgcgag gacgccggcg gcggcgccgg cggcaggaga caagaacctg cggttcctgc 1800
tgcagaaggt gctcaagcag agcgacgtcg gaaccctcgg ccgcatcgtg ctccccaaag 1860
tgaggcagtc acctccctat cgcggtcttc ttcctccatc cgcccgaccc atgcaccact 1920
tgcaattttc agctatatat gcatgcatcc acttgcaaat tttagtttct ccttgattcc 1980
tctttgatta gacaagatct atctttggat tgcgcctgta tgacctagct atgaatggat 2040
cttcctttct tttctgtaat ctttgtgttg atttcttctt gatttctcgg attgcaagtg 2100
tatctatcca tggatcttcc tgtctttctg ctgatcgatc ggttgcgact aaaaagtgtc 2160
tcttcttcca tggatggttt cagaaggaag cggagactca cctgccggag ctcaagacgg 2220
gggacggcat ctcgatcccc attgaggaca tcggcacatc tcaggtctgg agcatgcggt 2280
accggtaagt agtactcccg tcgatcgatt ttgggataaa tttaataata atattaaaca 2340
tgcatgcttt cattagtcca cttttatctt tttttaatta tctgtaggcc acatctagta 2400
aatgacatga tcactctgca tcactgtggg acacgcatgc tcctttccca attgggcaag 2460
tgggtcatat gaatacatat gatgggaatc cgggagcatc cgggagcaaa attgctttac 2520
acacgacggt tagtgcacga cgcttgcacg attctccctc cccaacaggg ctcctccaca 2580
cacggtggct gctggtgctt tttttaattg agaatcggtc gctgctggtg ctgccacgta 2640
tgaatcgttc atgcatcggc cgtgcaggca tcgtctatct agcaacgctc atccgggagc 2700
aaaattgctt tactgcgctc tctctatata tacgtgaacg ggcgggcatg cagaaatttt 2760
gttttcgcat cgtctcttgg ttcactcctt gttagcatca ctctagacag tatattatca 2820
taatgaagtg gtgtttctga taccatactt cctatttttc cgtcttcttc ttcttcttcc 2880
gaagtgtatc atatgttgct agttactccc tccgtaaact aatataagag cgtttagatt 2940
actattttag tgatctaaat gctcttatat tagtttacag agggagtaat tagctagagc 3000
tatcaaaatg agaaaaaaag aactagctag tttgtattct gcatgcatga taacaaattg 3060
ctggtacaat gatggctctt gatttttcat gcatgcagat tttggcccaa caacaagagc 3120
agaatgtatc ttctagagaa cactggtgag agaagagaag caaacttgcc tgatacaatt 3180
ttgaaaccaa atatgttgct agcttccctt gtgttcacaa aacgtttctt ctgcttgacc 3240
ttgtaggtga ctttgttcgg tccaatgagc tgcaggaggg tgatttcatc gtgctttact 3300
ctgatgtcaa gtcaggcaaa tatgtaatga tcataccctt cccgttaatt catttccatg 3360
aaatgatcgg tttacatcca tatgcatgtt gcgaactgat cagttgctgc cattttggga 3420
gcgttggcat ataattatca caatatatac tatcccgtcc aaaaatataa tacggattat 3480
tatgtactat gaggtcaacg ttttaaaact ttggcctctt gtatgtgcgt aagtatatgc 3540
atggatattg tgcatatatt aatagcaata acatcgtatt catcttgcaa aatagtttta 3600
tatcatgcat atctccctat tctttatgca tattatgagg tcaaaatcaa gtgttggaga 3660
ctgaaaagtc agatgtctat gtcttgtatt ctgtgaggga tggagtatat aggactgttt 3720
gtctttggag tttggatatc taattaagta cgtactagca gaactatctg tgttcttgtt 3780
gtgcatttga atgaattgtc tatacatcag ctaatctcag cctttctgtg taccaccagc 3840
tgatacgcgg cgtgaaggta agagctcaac aggatctagc caagcacaag aatgccagtc 3900
cagagaaagg tggggcgtcc gacgtgaagg cgggcggaga agacggtggt tgcaaagaga 3960
agtctccgca cggtgtccgg cgatctcgcc aggaagccag ctccatgaac cagatggcgg 4020
tgagcatctg aaatgagcag gctcgccgtc cgatccccca ttgaagacta cttaattagc 4080
tagctcaagt atacctgcta atgatgatca aatcgatctc ccgttctatg atccgtgctt 4140
ccgtgtactg ccgtagccct agttagggat gatgatacta aagtagctat cggtttcttg 4200
gtcagatgtg acgctgaaga atggcat 4227
<210>3
<211>3951
<212>DNA
＜213〉Triticum common wheat west farming 979 (Triticum aestivum L cv Xinong 979)
<400>3
atggacgcct ccgccggctc gtcgccgccg cggcactcgc agggggacgc gccgaggcgc 60
ggcgggccgc accgcgggaa gggccccgcg gtggagatcc ggcagggaga ggacgacttc 120
atgttcgcgc aggatacctt cccggccctc ccggacttcc cttgcctctc ctcgccgtcg 180
agctccacct tctcctcctc ctcgtcttcc aactcctcca gcgccttcgc tgccccagcg 240
ggagcgggcg ggcgcggggc cgagggggcg cgcggcgagc cgtccgagcc tgcagcggcc 300
ggggacggga tggacgacct ctccgacatc gaccacctgc tcgacttcgc gtccatcaac 360
gacgacgtcc cctgggacga cgagccgctc ttccccgacg tcgggatgat gctggaggat 420
gtcatctccg agcagcagct ccagcagcct ccggcgggcc acggcacggc cgggagaacg 480
gcgtcgcatg cggcctctgg tggaggagag gatgccttca tgggtggcgg cggcacgggg 540
agcgcggcgg acgacctgcc gctgttcttc atggagtggc tcaagaacaa ccgcgactgc 600
atctcggccg aggacctccg cagcatccgc ctccgtcgat ccaccatcga ggccgcggcc 660
gcgcgcctcg gtggggggcg ccagggcacc atgcagctgc tcaagctcat cctcacctgg 720
gtgcagaacc accacctgca gaagaagcgc ccccgcgtcg gcgccatgga tcaggaggcg 780
ccgccggcag gaggccagct ccctagcccc ggcgcaaacc ccggctacga attccccgcg 840
gagacgggtg ccgccgctgc cacatcatgg atgccctacc aggccttctc gccaactgga 900
tcctacggcg gcgaggcgat ctacccattc cagcagggct gcagcactag cagcgtggcc 960
gtgagcagcc agccgttctc cccgccggcg gcgcccgaca tgcacgccgg ggcctggccc 1020
ctgcagtacg cggcgttcgt cccagctgga gccacatccg caggcactca aacatacccg 1080
atgccgccgc cgggggccgt gccgcagccg ttcgcggccc ccggattcgc cgggcagttc 1140
ccgcagcgga tggagccggc ggcgaccagg gaggcccgga agaagcggat ggcgaggcag 1200
cggcgcttgt cgtgcctgca gcagcagcgg agccagcagc tgaatctgag ccagatccaa 1260
accggcggct tccctcaaga gcaatccccc cgcgcggcgc actcggcgcc ggtcacgccg 1320
ccgtcgtccg gctggggagg cctctggacg caacaggccg tccagggcca gctcatggtc 1380
caggtcccga atccgctgtc gacgaagtcc aattcctcaa ggcagaagca gcaaaaaccc 1440
tcgccggacg cagcagcgag gccgccctcc ggcggcgccg ccacgccgca tcgcccgggg 1500
caggcgtcgg cttccaataa gcagcggcag caggtgcatg catgcacgaa cacctcttgc 1560
catccatcca tcgatcgcca tcccgcatag aatcacaagc cattgctccc caaataaagt 1620
gtgcgtgctt cgtaatggac gcacatcgct gtccagcgat aggatatgca tgcgcgcgtg 1680
cgtgtctgtt ttacctgcac ggtgaattgc gtttctcaac gaggttccgt gcatgcgcgc 1740
agggtgcgag gacgccggcg gcggcgccgg cggcaggaga caagaacctg cggttcctgc 1800
tgcagaaggt gctcaagcag agcgacgtcg gaaccctcgg ccgcatcgtg ctccccaaag 1860
tgaggcagtc acctccctat cgcggtcttc ttcctccatc cgcccgaccc atgcaccact 1920
tgcaattttc agctatatat gcatgcatcc acttgcaaat tttagtttct ccttgattcc 1980
tctttgatta gacaagatct atctttggat tgcgcctgta tgacctagct atgaatggat 2040
cttcctttct tttctgtaat ctttgtgttg atttcttctt gatttctcgg attgcaagtg 2100
tatctatcca tggatcttcc tgtctttctg ctgatcgatc ggttgcgact aaaaagtgtc 2160
tcttcttcca tggatggttt cagaaggaag cggagactca cctgccggag ctcaagacgg 2220
gggacggcat ctcgatcccc attgaggaca tcggcacatc tcaggtctgg agcatgcggt 2280
accggtaagt agtactcccg tcgatcgatt ttgggataaa tttaataata atattaaaca 2340
tgcatgcttt cattagtcca cttttatctt tttttaatta tctgtaggcc acatctagta 2400
aatgacatga tcactctgca tcactgtggg acacgcatgc tcctttccca attgggcaag 2460
tgggtcatat gaatacatat gatgggaatc cgggagcatc cgggagcaaa attgctttac 2520
tgcgctctct ctatatatac gtgaacgggc gggcatgcag aaattttgtt ttcgcatcgt 2580
ctcttggttc actccttgtt agcatcactc tagacagtat attatcataa tgaagtggtg 2640
tttctgatac catacttcct atttttccgt cttcttcttc ttcttccgaa gtgtatcata 2700
tgttgctagt taattagcta gagctatcaa aatgagaaaa aaagaactag ctagtttgta 2760
ttctgcatgc atgataacaa attgctggta caatgatggc tcttgatttt tcatgcatgc 2820
agattttggc ccaacaacaa gagcagaatg tatcttctag agaacactgg tgagagaaga 2880
gaagcaaact tgcctgatac aattttgaaa ccaaatatgt tgctagcttc ccttgtgttc 2940
acaaaacgtt tcttctgctt gaccttgtag gtgactttgt tcggtccaat gagctgcagg 3000
agggtgattt catcgtgctt tactctgatg tcaagtcagg caaatatgta atgatcatac 3060
ccttcccgtt aattcatttc catgaaatga tcggtttaca tccatatgca tgttgcgaac 3120
tgatcagttg ctgccatttt gggagcgttg gcatataatt atcacaatat atactatccc 3180
gtccaaaaat ataatacgga ttattatgta ctatgaggtc aacgttttaa aactttggcc 3240
tcttgtatgt gcgtaagtat atgcatggat attgtgcata tattaatagc aataacatcg 3300
tattcatctt gcaaaatagt tttatatcat gcatatctcc ctattcttta tgcatattat 3360
gaggtcaaaa tcaagtgttg gagactgaaa agtcagatgt ctatgtcttg tattctgtga 3420
gggatggagt atataggact gtttgtcttt ggagtttgga tatctaatta agtacgtact 3480
agcagaacta tctgtgttct tgttgtgcat ttgaatgaat tgtctataca tcagctaatc 3540
tcagcctttc tgtgtaccac cagctgatac gcggcgtgaa ggtaagagct caacaggatc 3600
tagccaagca caagaatgcc agtccagaga aaggtggggc gtccgacgtg aaggcgggcg 3660
gagaagacgg tggttgcaaa gagaagtctc cgcacggtgt ccggcgatct cgccaggaag 3720
ccagctccat gaaccagatg gcggtgagca tctgaaatga gcaggctcgc cgtccgatcc 3780
cccattgaag actacttaat tagctagctc aagtatacct gctaatgatg atcaaatcga 3840
tctcccgttc tatgatccgt gcttccgtgt actgccgtag ccctagttag ggatgatgat 3900
actaaagtag ctatcggttt cttggtcaga tgtgacgctg aagaatggca t 3951
<210>4
<211>688
＜212〉protein
＜213〉excellent 9507 (Triticum aestivum L cv Zhongyou 9507) in the Triticum common wheat
<400>4
Met Asp Ala Ser Ala Gly Ser Ser Pro Pro Arg His Ser Gln Gly Asp
1 5 10 15
Ala Pro Arg Arg Gly Gly Pro His Arg Gly Lys Gly Pro Ala Val Glu
20 25 30
Ile Arg Gln Gly Glu Asp Asp Phe Met Phe Ala His Asp Thr Phe Pro
35 40 45
Ala Leu Pro Asp Phe Pro Cys Leu Ser Ser Pro Ser Ser Ser Thr Phe
50 55 60
Ser Ser Ser Ser Ser Ser Asn Ser Ser Ser Ala Phe Ala Ala Pro Ala
65 70 75 80
Gly Ala Gly Gly Arg Gly Ala Glu Gly Ala Arg Gly Glu Pro Ser Glu
85 90 95
Pro Ala Ala Ala Gly Asp Gly Met Asp Asp Leu Ser Asp Ile Asp His
100 105 110
Leu Leu Asp Phe Ala Ser Ile Asn Asp Asp Val Pro Trp Asp Asp Glu
115 120 125
Pro Leu Phe Pro Asp Val Gly Met Met Leu Glu Asp Val Ile Ser Glu
130 135 140
Gln Gln Leu Gln Gln Pro Pro Ala Gly His Gly Thr Ala Gly Arg Thr
145 150 155 160
Ala Ser His Ala Ala Ser Gly Gly Gly Glu Asp Ala Phe Met Gly Gly
165 170 175
Gly Gly Thr Gly Ser Ala Ala Asp Asp Leu Pro Leu Phe Phe Met Glu
180 185 190
Trp Leu Lys Asn Asn Arg Asp Cys Ile Ser Ala Glu Asp Leu Arg Ser
195 200 205
Ile Arg Leu Arg Arg Ser Thr Ile Glu Ala Ala Ala Ala Arg Leu Gly
210 215 220
Gly Gly Arg Gln Gly Thr Met Gln Leu Leu Lys Leu Ile Leu Thr Trp
225 230 235 240
Val Gln Asn His His Leu Gln Lys Lys Arg Pro Arg Val Gly Ala Met
245 250 255
Asp Gln Glu Ala Pro Pro Ala Gly Gly Gln Leu Pro Ser Pro Gly Ala
260 265 270
Asn Pro Gly Tyr Glu Phe Pro Ala Glu Thr Gly Ala Ala Ala Ala Thr
275 280 285
Ser Trp Met Pro Tyr Gln Ala Phe Ser Pro Thr Gly Ser Tyr Gly Gly
290 295 300
Glu Ala Ile Tyr Pro Phe Gln Gln Gly Cys Ser Thr Ser Ser Val Ala
305 310 315 320
Val Ser Ser Gln Pro Phe Ser Pro Pro Ala Ala Pro Asp Met His Ala
325 330 335
Gly Ala Trp Pro Leu Gln Tyr Ala Ala Phe Val Pro Ala Gly Ala Thr
340 345 350
Ser Ala Gly Thr Gln Thr Tyr Pro Met Pro Pro Pro Gly Ala Val Pro
355 360 365
Gln Pro Phe Ala Ala Pro Gly Phe Ala Gly Gln Phe Pro Gln Arg Met
370 375 380
Glu Pro Ala Ala Thr Arg Glu Ala Arg Lys Lys Arg Met Ala Arg Gln
385 390 395 400
Arg Arg Leu Ser Cys Leu Gln Gln Gln Arg Ser Gln Gln Leu Asn Leu
405 410 415
Ser Gln Ile Gln Thr Gly Gly Phe Pro Gln Glu Gln Ser Pro Arg Ala
420 425 430
Ala His Ser Ala Pro Val Thr Pro Pro Ser Ser Gly Trp Gly Gly Leu
435 440 445
Trp Thr Gln Gln Ala Val Gln Gly Gln Leu Met Val Gln Val Pro Asn
450 455 460
Pro Leu Ser Thr Lys Ser Asn Ser Ser Arg Gln Lys Gln Gln Lys Pro
465 470 475 480
Ser Pro Asp Ala Ala Ala Arg Pro Pro Ser Gly Gly Ala Ala Thr Pro
485 490 495
His Arg Pro Gly Gln Ala Ser Ala Ser Asn Lys Gln Arg Gln Gln Gly
500 505 510
Ala Arg Thr Pro Ala Ala Ala Pro Ala Ala Gly Asp Lys Asn Leu Arg
515 520 525
Phe Leu Leu Gln Lys Val Leu Lys Gln Ser Asp Val Gly Thr Leu Gly
530 535 540
Arg Ile Val Leu Pro Lys Lys Glu Ala Glu Thr His Leu Pro Glu Leu
545 550 555 560
Lys Thr Gly Asp Gly Ile Ser Ile Pro Ile Glu Asp Ile Gly Thr Ser
565 570 575
Gln Val Trp Ser Met Arg Tyr Arg Phe Trp Pro Asn Asn Lys Ser Arg
580 585 590
Met Tyr Leu Leu Glu Asn Thr Gly Asp Phe Val Arg Ser Asn Glu Leu
595 600 605
Gln Glu Gly Asp Phe Ile Val Leu Tyr Ser Asp Val Lys Ser Gly Lys
610 615 620
Tyr Leu Ile Arg Gly Val Lys Val Arg Ala Gln Gln Asp Leu Ala Lys
625 630 635 640
His Lys Asn Ala Ser Pro Glu Lys Gly Gly Ala Ser Asp Val Lys Ala
645 650 655
Gly Gly Glu Asp Gly Gly Cys Lys Glu Lys Ser Pro His Gly Val Arg
660 665 670
Arg Ser Arg Gln Glu Ala Ser Ser Met Asn Gln Met Ala Val Ser Ile
675 680 685
<210>5
<211>18
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>5
tgctcctttc ccaattgg 18
<210>6
<211>19
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>6
accctcctgc agctcattg 19
Claims (10)
1, a kind of wheat ear germinating resistance related gene has one of following nucleotide sequence:
1) SEQ ID № in the sequence table: 2 nucleotide sequence;
2) with sequence table in SEQ ID №: 2 dna sequence dna has 90% above homology, and the identical function protein DNA sequence of encoding;
3) under the rigorous condition of height can with the SEQ ID № in the sequence table: the nucleotide sequence of the 2 dna sequence dnas hybridization that limit.
2, the recombinant expression vector, transgenic cell line or the engineering bacteria that contain the described wheat ear germinating resistance related gene of claim 1.
3, a kind of assisting sifting ear germinating resistance method of wheat, be to detect the nucleotide sequence that whether has inserted 193bp in the 3rd intron of Vp-1B gene of wheat to be measured, inserted the nucleotide sequence of 193bp in the 3rd intron as the Vp-1B gene of this wheat to be measured, then this wheat to be measured is candidate's ear germinating resistance wheat.
4, method according to claim 3 is characterized in that: detect the nucleotide sequence that whether has inserted 193bp in the 3rd intron of Vp-1B gene of wheat to be measured with PCR method in the described method.
5, method according to claim 4, it is characterized in that: described PCR method is a template with the genomic dna of wheat to be measured, use a pair of primer of forming by the nucleotide sequence of sequence in the sequence table 5 and sequence 6 to carry out pcr amplification, if there is size to be that the band of 845bp, then described wheat to be measured are candidate's ear germinating resistance wheat in the pcr amplification product that obtains.
6, method according to claim 5 is characterized in that: the method for described detection amplified production is amplified production to be carried out 2.0% agarose detection, 130V voltage electrophoresis 1.5 hours.
7, according to claim 3,4,5 or 6 described methods, it is characterized in that: described wheat to be measured is the white wheat kind.
8, a kind of method of assisting sifting fringe germination sensitive wheat, be to detect to have or not in the 3rd intron of Vp-1B gene of wheat to be measured to insert and deletion mutantion, do not insert and deletion mutantion as having in the 3rd intron of the Vp-1B gene of this wheat to be measured, then this wheat to be measured is candidate's fringe germination sensitive wheat kind;
Described insertion sports the sudden change of inserting the nucleotide sequence of 193bp in the 3rd intron of Vp-1B gene, and described deletion mutantion is the sudden change of the nucleotide sequence of disappearance 83bp in the 3rd intron of Vp-1B gene.
9, method according to claim 8 is characterized in that: in the described method, detect with PCR method to have or not in the 3rd intron of Vp-1B gene of wheat to be measured and insert and deletion mutantion;
Described PCR method is a template for the genomic dna with wheat to be measured, use a pair of primer of forming by the nucleotide sequence of sequence in the sequence table 5 and sequence 6 to carry out pcr amplification, if there be not the band of size for 845bp and 569bp in the pcr amplification product that obtains, then described wheat breed to be measured is candidate's fringe germination sensitive wheat varieties.
10, according to Claim 8 or 9 described methods, it is characterized in that: described wheat to be measured is the white wheat kind.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610089191 CN1904050A (en) | 2006-08-08 | 2006-08-08 | Wheat ear germinating resistance related gene and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610089191 CN1904050A (en) | 2006-08-08 | 2006-08-08 | Wheat ear germinating resistance related gene and its application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1904050A true CN1904050A (en) | 2007-01-31 |
Family
ID=37673415
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200610089191 Pending CN1904050A (en) | 2006-08-08 | 2006-08-08 | Wheat ear germinating resistance related gene and its application |
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| Country | Link |
|---|---|
| CN (1) | CN1904050A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103004584A (en) * | 2012-10-22 | 2013-04-03 | 西北农林科技大学 | Method for rapidly cultivating wheat pre-harvest sprouting material from complex genotype group |
| CN105483136A (en) * | 2016-01-04 | 2016-04-13 | 安徽农业大学 | wheat pre-harvest sprouting tolerance gene TaZFP18 and application thereof |
| CN105671060A (en) * | 2016-04-26 | 2016-06-15 | 杨燕 | Wheat pre-harvest sprouting resistance related gene and application thereof |
| CN105861667A (en) * | 2016-04-20 | 2016-08-17 | 中国农业科学院作物科学研究所 | Molecular marker and specific primer for identifying wheat grain germination traits and application thereof |
| CN106011301A (en) * | 2016-08-03 | 2016-10-12 | 中国科学院成都生物研究所 | Primer for throughput identification of wheat seed major dormancy genes, and kit and method thereof |
-
2006
- 2006-08-08 CN CN 200610089191 patent/CN1904050A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103004584A (en) * | 2012-10-22 | 2013-04-03 | 西北农林科技大学 | Method for rapidly cultivating wheat pre-harvest sprouting material from complex genotype group |
| CN103004584B (en) * | 2012-10-22 | 2014-10-29 | 西北农林科技大学 | Method for rapidly cultivating wheat pre-harvest sprouting material from complex genotype group |
| CN105483136A (en) * | 2016-01-04 | 2016-04-13 | 安徽农业大学 | wheat pre-harvest sprouting tolerance gene TaZFP18 and application thereof |
| CN105483136B (en) * | 2016-01-04 | 2019-01-04 | 安徽农业大学 | A kind of wheat anti growing out gene TaZFP18 and its application |
| CN105861667A (en) * | 2016-04-20 | 2016-08-17 | 中国农业科学院作物科学研究所 | Molecular marker and specific primer for identifying wheat grain germination traits and application thereof |
| CN105861667B (en) * | 2016-04-20 | 2019-10-11 | 中国农业科学院作物科学研究所 | For identifying molecular labeling and primer pair and their application of Wheat Grain Germination character |
| CN105671060A (en) * | 2016-04-26 | 2016-06-15 | 杨燕 | Wheat pre-harvest sprouting resistance related gene and application thereof |
| CN105671060B (en) * | 2016-04-26 | 2019-07-23 | 杨燕 | Wheat ear germinating resistance related gene and its application |
| CN106011301A (en) * | 2016-08-03 | 2016-10-12 | 中国科学院成都生物研究所 | Primer for throughput identification of wheat seed major dormancy genes, and kit and method thereof |
| CN106011301B (en) * | 2016-08-03 | 2019-08-23 | 中国科学院成都生物研究所 | A kind of primer, kit and its method of throughput assays wheat seed main effect dormant gene |
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Open date: 20070131 |