CN107699630A - Chain molecular labeling and its application in breeding with wheat resistance genes Pm21 - Google Patents
Chain molecular labeling and its application in breeding with wheat resistance genes Pm21 Download PDFInfo
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- 241000122204 Dasypyrum villosum Species 0.000 abstract description 26
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Abstract
Application the invention discloses the molecular labeling chain with wheat resistance genes Pm21 and its in breeding.25 molecular labelings are from containing the minimum fragment translocation line NAU427 of Pm21 mildew-resistance gene wheat Haynaldia villosas, and Pm21 gene close linkages, available for molecular marker assisted selection, improve the efficiency of selection and accuracy rate of disease-resistant plant.
Description
Technical field
The invention belongs to molecular genetic breeding field, be related to the molecular labeling chain with wheat resistance genes Pm21 and its
Application in breeding.
Background technology
Wheat is global important cereal crops, and there is 35%-40% people in the whole world using wheat as staple food.Wheat white powder
Disease is one of worldwide important disease, and the general time of falling ill can cause 13-34% production loss, seriously fall ill the time
More than 50% production loss can be caused.It is the most economic, safely effectively approach to cultivate high resist powdery mildew of wheat kind.Closely
Nian Lai, as the development of molecular marking technique, the structure of High Density Molecular genetic map, molecular mark are excellent in wheat
More and more important effect has been played in benign shape election effects.Excavate the germ plasm resource containing mildew-resistance gene, exploitation with
The chain molecular labeling of disease-resistant gene, it is the basis for carrying out wheat anti-powdery mildew genetic breeding.
At present, 58 gene locis have excavated 85 mildew-resistance genes from wheat and its nearly edge species, but these
Gene is largely race specific resistance, be easy to lose its disease resistance (Wiersma AT, Pulman JA, Brown LK,
Cowger C,Olson EL.Identification of Pm58from Aegilops tauschii. Theor Appl
Genet,2017,130:1123-1133).It is high anti-that powdery mildew resistance gene Pm 21 from haynaldia villosa shows powdery mildew wide spectrum.Cluster
Dirty wheat (Haynaldia villosa) has 7 pairs of chromosomes (2n=2x=14, VV), and each pair chromosome includes two homologous dyeing
Body, this 7 chromosomes are respectively 1V, 2V, 3V, 4V, 5V, 6V, 7V.The high anti-white powder of wide spectrum on the 6V galianconism of haynaldia villosa be present
Ospc gene Pm21, the gene can resist more than 120 powdery mildew biological strains (Huang XQ, Hsam SLK, Zeller both at home and abroad
FJ.Identification of powdery mildew resistance genes in common wheat(Triticum
aestivum L em Thell).9.Cultivars,land races and breeding lines grown in
China.Plant Breeding, 1997,116:233-238).Agricultural University Of Nanjing's cytogenetics has formulated wheat-tuft
Wheat translocation line T6VS6AL (Chen PD, Qi LL, Zhou B, Zhang SZ, Liu DJ.Development and
molecular cytogenetic analysis of wheat-Haynaldia villosa 6VS/6AL
translocation lines specifying resistance to powdery mildew.TAG,1995,(91):
1125-1128), the haynaldia villosa 6VS chromosome arms containing Pm21 genes are replaced into common wheat 6AL chromosome arms, made common small
Wheat obtains the resistance of wide spectrum to powdery mildew, and more than 30 individual kinds are widely applied kind in China using the translocation line as parent's seed selection
Plant.While 6VS chromosome arms import general wheat background introducing powdery mildew resistance gene Pm 21,6VS is inevitably carried
Other genes on chromosome arm.Agricultural University Of Nanjing's cytogenetics has formulated the small fragment for carrying powdery mildew resistance gene Pm 21
Translocation line NAU418, and molecular labeling CINAU273, CINAU274, the CINAU275 of traceable transposition fragment are developed,
CINAU15(Chen PD,You CF,Hu Y,Chen SW,Zhou B,Cao AZ,Wang X.Radiation-induced
translocations with reduced Haynaldia villosa chromatin at the Pm21locus for
powdery mildew resistance in wheat.Mol Breed 2013,31:477-484).The top that NAU418 is carried
Although end transposition fragment is obviously reduced compared with the transposition fragment than whole arm T6VS6AL, the transposition fragment is still larger,
It can detect to obtain by in-situ hybridization method.Further initiative carries the minimum fragment translocation line of Pm21 genes, the easy bit slice of exploitation
It may be either that disease-resistant wheat genetic breeding provides new germ plasm resource with the molecular labeling of Pm21 gene linkages in section, can also be point
Sub- marker assisted selection provides available molecular labeling.
The content of the invention：
The present invention seeks to the drawbacks described above for prior art, there is provided one contains the small of powdery mildew resistance gene Pm 21
The minimum fragment translocation line NAU427 of wheat-haynaldia villosa and with the molecule mark with Pm21 gene close linkages contained by its transposition fragment
Note.NAU427 can be used for wheat anti-powdery mildew genetic breeding as germ plasm resource, avoid the redundancy of lots of genes on 6VS chain；
It can be used for molecular marker assisted selection in NAU427 transposition fragment with the molecular labeling of Pm21 gene close linkages, improve breeding
Efficiency.
The purpose of the present invention can be achieved through the following technical solutions：
The chain molecular labeling with wheat resistance genes Pm21, it is characterised in that selected from CINAU1692, CINAU1693,
CINAU1694、CINAU1695、CINAU1696、CINAU1697、CINAU1698、 CINAU1699、CINAU1700、
CINAU1701、CINAU1702、CINAU1703、CINAU1704、 CINAU1705、CINAU1706、CINAU1707、
CINAU1708、CINAU1709、CINAU1710、 CINAU1711、CINAU1712、CINAU1713、CINAU1714、
Any one in CINAU1715, CINAU1716；Its primer sequence is as shown in table 1：
Table 1
。
Primer of the present invention with molecular labeling chain disease-resistant gene Pm21, sequence such as SEQ ID NO.1-SEQ
Shown in ID NO.50.
Application of the primer of the present invention in Wheat Germplasm Resources of the identification containing Pm21 genes, strain and kind.
Application of the primer of the present invention in wheat breeding for disease resistance.
The disease-resistant translocation line NAU427 of minimum fragment containing resistant gene to wheat powdery mildew Pm21, is preserved in Chinese Typical Representative culture
Thing collection, preservation date 2017.3.17, preserving number are CCTCC P201707, and preservation address is Wuhan, China university.
Minimum radiation of the fragment translocation line NAU427 from T6VS6AL whole arm translocations system 92R137 of the powdery-mildew-resistance wheat-haynaldia villosa
Offspring.
The disease-resistant translocation line NAU427 of minimum fragment containing resistant gene to wheat powdery mildew Pm21 of the present invention is in wheat
Application in breeding for disease resistance.
Beneficial effects of the present invention are：
(1) present invention obtains wheat-haynaldia villosa pole Small piece transposition from the Irradiated Progenies screening of T6VS6AL whole arm translocations system
It is that NAU427 contains resistant gene to wheat powdery mildew Pm21, high resist powdery mildew of wheat, the anti-white powder of wheat can be used for as germ plasm resource
Sick genetic breeding, avoid the unfavorable chain of 6VS whole arm translocation fragment bulk redundancy genes.(2) present invention is developing with disease-resistant base
Because molecular labeling chain Pm21 can be used for molecular marker assisted selection breeding, wheat breeding for disease resistance efficiency is greatly improved.(3) originally
It is to be lacked for special insertion in haynaldia villosa genome sequence to invent 25 molecular labelings chain with Pm21 wherein 24 provided
Unsceptered point design, can the specific band that goes out gradually to penetrate on NAU427 haynaldia villosa chromatin of specific amplified, can be by 1% fine jade
Sepharose electrophoresis fast detects, the mark that wherein CINAU1704 is developed based on gene intron difference in size, and PCR amplifications are anti-
Amplified production is utilizing non-denaturing polyacrylamide gel (acrylamide after answering：Methene acrylamide=39:1) detect, electrophoresis
The Rapid identification of molecular marker assisted selection easy to detect.
Brief description of the drawings
The seedling stage (A) and Adult plant (B) Resistance Identification of the minimum fragment translocation line NAU427 of Fig. 1 and susceptible control China spring；
The minimum fragment translocation line NAU427 of Fig. 2 and its in situ hybridization picture (A figures) and molecular labeling mirror for compareing NAU418
Determine picture (B figures)；
1：Maker；2：CS (is free of Pm21 disease-resistant genes)；3：VV (haynaldia villosa, contains Pm21 disease-resistant genes)；4：ABV (people
Work synthetic wheat, contains Pm21)；5：T6VS6AL (contains Pm21 disease-resistant genes)；6：NAU418 (contains Pm21 disease-resistant genes
Small piece transposition system)；7：NAU427
The disease-resistant material NAU427 of Fig. 3 and sense powdery mildew kind Yangmai No.158, the backcross progeny plant of stone 4185 disease resistance are reflected
Fixed and molecular labeling CINAU1704 identifications；
Y158：Yangmai No.158；Y158(R)/SI+：NAU427 and the disease-resistant material in Y158 backcross progeny segregating populations.Stone
4185(R)/SI+：NAU427 and the disease-resistant material in the backcross progeny segregating population of stone 4185.
Amplification of 25 linked markers of Fig. 4 in various materials
1：Maker；2：CS (is free of Pm21 disease-resistant genes)；3：VV (haynaldia villosa, contains Pm21 disease-resistant genes)；4：ABV (people
Work synthetic wheat, contains Pm21)；5：T6VS6AL (contains Pm21 disease-resistant genes)；6：NAU418 (contains Pm21 disease-resistant genes
Small piece transposition system)；7：NAU427
Biological sample preservation information
Wheat seed NAU427, Classification And Nomenclature are Triticum Wheat Species NAU427 (Triticum aestivum
NAU427), it is preserved in China typical culture collection center, preservation date is on March 17th, 2017, preserving number CCTCC
NO:P201707, preservation address are Wuhan, China university.
Embodiment
Embodiment 1. carries the minimum fragment translocation line NAU427 of wheat-haynaldia villosa of Pm21 genes initiative
(1) in academy of agricultural sciences of Jiangsu Province with 1600 dosage60The whole arms of T6VS6AL of 2-3 days before CO gamma Rays are bloomed
Translocation line 92R137 tassel, emasculation is carried out on the day of to irradiated tassel, with the normal pollen of China spring to it after 2-3 days
Pollination, obtain structural variant M1 (Chen PD, You CF, Hu Y, Chen SW, Zhou B, Cao AZ, Wang X (2013)
Radiation-induced translocations with reduced Haynaldia villosa chromatin at
the Pm21locus for powdery mildew resistance in wheat.Mol Breed 31:477-484:)。
(2) field is arrived in the plantation of M1 seeds, and seedling and Adult plant carry out powdery mildew inoculated identification, screen high resist powdery mildew of wheat
Plant, the maturity period harvest seed.
(3) to the seed clip tip of a root harvested in disease-resistant plant, in situ hybridization is carried out by probe of haynaldia villosa genomic DNA
Experiment, detect the size that the H. villosa chromosome in disease-resistant plant gradually oozes fragment.An individual plant is screened from disease-resistance population, should
The hybridization signal of haynaldia villosa genome is can't detect on the root tip cell chromosome of material, thus it is speculated that there occurs pole Small piece transposition gradually
Event is oozed, and gradually oozes carrying Pm21 genes in fragment.
(4) 6VS specific molecular marker CINAU273, CINAU274, the CINAU275 developed in NAU418 is utilized,
CINAU15 the material is entered performing PCR amplification, wherein with mark CINAU273 it is amplifiable go out 6VS on specific molecular marker, enter one
The chromosome segment for carrying Pm21 genes is gradually permeated with step clearly disease-resistant material.
(5) the above-mentioned disease-resistant, Small piece transposition obtained gradually oozes structural variant and China spring is repeatedly returned, each to return
Hand over and all carry out Disease Resistance Identification and molecular labeling CINAU273 Amplification Analysis from generation to generation, obtain one in the time of infertility high anti-wheat
The stabilizing material of powdery mildew, is named as NAU427.The material is preserved in China typical culture collection center, and preservation date is
2017.3.17, preserving number is CCTCC NO:P201707.
The minimum disease-resistant effect analysis of fragment translocation line NAU427 of the wheat-haynaldia villosa of embodiment 2.
(1), will in order to analyze the disease-resistant effect of transposition fragment in the minimum fragment translocation line NAU427 of wheat-haynaldia villosa
NAU427 and sense powdery mildew commercial variety stone 4185, the Yangmai No.158 of planted in different ecological areas carry out hybridization and obtain F1, then respectively with popularization
Kind stone 4185 and Yangmai No.158 are that recurrent parent is repeatedly returned, each to carry out Disease Resistance Identification and molecular labeling from generation to generation
CINAU1704 is identified, disease-resistant plant is carried out into selfing harvest seed BC4F1 in backcrossing BC4 generations.
(2) to selfed seed plantation into Agricultural University Of Nanjing Jiangpu farm vinyl house, row long 1.2m, line-spacing 20cm,
Often row broadcasts 10, and powder mildew resistance identification is carried out to individual plant in seedling stage and Adult plant, and at least 3 are carried out in late March and early April
Secondary Disease Resistance Identification.
(3) DNA of disease-resistant plant and disease plant is extracted, transposition fragment presence is carried out using molecular labeling CINAU1704
In no analysis, the plant for finding that 6VS specific bands can be amplified is disease-resistant plant, it is impossible to amplifies 6VS specific bands
Plant is disease plant.
The above results show：Haynaldia villosa 6VS translocation chromosome fragments are carried in NAU427, the fragment carries Pm21 genes.
The disease-resistant effect for the minimum transposition fragment that NAU427 is carried can stablize heredity.
The molecular markers development (1) chain with Pm21 in the minimum fragment translocation line NAU427 of the wheat-haynaldia villosa of embodiment 3.
The exploitation of the molecular labeling chain with Pm21 in NAU427
The haynaldia villosa transposition fragment that 1.1 NAU427 gradually ooze and the determination of false bromegrass and rice synteny section
NAU418 and NAU427 contains Pm21 genes, and in situ hybridization result shows that NAU418 transposition fragment compares NAU427
Transposition fragment it is big.In order to develop the chain molecular labeling in NAU427 transposition fragments with Pm21 genes, it is extracted first
4 marks CINAU273, CINAU274, CINAU275 of 6VS specific bands are amplified in Small piece transposition system NAU418,
CINAU15 est sequence.By this 4 est sequences respectively with false bromegrass (http://plants.ensembl.org/
) and rice (http Brachypodium_distachyon/Info/Index://plants.ensembl.org/Oryza_
Sativa/Info/Index genome sequence) compares, and comparison result finds that this 4 est sequences contaminate in the Article 3 of false bromegrass
Colour solid is consistent with putting in order on the 2nd article of chromosome of rice, illustrates haynaldia villosa and false bromegrass and the genome sequence of rice
There is preferable co-linear relationship in the section.Gene BRADI3G03270 on false bromegrass Article 3 chromosome is further extracted to arrive
Gene BRADI3G04940 gene order, the false bromegrass gene order extracted are searched haynaldia villosa genome sequence, obtained respectively
Obtain and extract the sequence of haynaldia villosa homologous gene.
The determination in special insertion/deletion site in 1.2 haynaldia villosa gene orders
The sequence of the haynaldia villosa homologous gene extracted with China spring 6A, corresponds to homologous gene on 6B and 6D chromosomes respectively
Sequence (https://urgi.versailles.inra.fr/blast/) Multiple Sequence Alignment analysis is carried out, if some tuft
One section of special insertion/deletion section, and the insertion/deletion section be present because being compared with 6A, 6B, 6D homologous gene in McGee
In China spring 6A, 6B and 6D sequence indifferences, then drawn according to amplification of the haynaldia villosa sequences Design for the insertion/deletion site
Thing.Primer utilizeshttp://primer3.ut.eePhotographing On-line, by Shanghai, Ying Jun Bioisystech Co., Ltd synthesizes.Utilize one
Series of cell material is verified whether primer designed by detection can amplify haynaldia villosa specific band, if in NAU427
In amplify the specific band of haynaldia villosa, then successfully develop the molecular labeling chain with Pm21.
The exploitation marked in 1.3 NAU427 with Pm21 linkage molecules
(1) DNA extraction：With haynaldia villosa (2n=2x=14, VV) of the CTAB methods extraction containing Pm21 genes, artificial conjunction
Into the hard cluster wheat (AABBVV) of wheat, wheat-haynaldia villosa whole arm translocation system T6VS6AL, top translocation line NAU418, minimum fragment
Translocation Materials NAU427 and the common wheat China spring (AABBDD) for not containing Pm21 genes.
(2) PCR programs：Enter performing PCR amplification, statistical analysis by template of the genomic DNA of each Cytologic material of extraction
Polymorphism, exploitation and the molecular labeling of Pm21 close linkages..PCR reagent forms in the reaction system of PCR amplifications：1μL DNA
Template (20-100ng), 1.0 μ 10 × PCR of L buffer, 0.8 μ L dNTP, left and right primer each 0.2 μ L, 0.1 μ L Taq DNA
Polymerase, 6.7 μ L ddH2O.PCR programs are：94 DEG C of pre-degenerations 3 minutes；Then 94 DEG C are denatured 30 seconds, 57 DEG C of renaturation 45
Second, 72 DEG C extend 1 minute, 35 circulations；Last 72 DEG C extend 10 minutes；10 DEG C of preservations.Amplified production removes after pcr amplification reaction
CINAU1704 is marked to utilize non-denaturing polyacrylamide gel (acrylamide：Methene acrylamide=39:1) electrophoresis is examined
Survey, remaining mark is detected with 1% agarose gel electrophoresis.
(3) determination in NAU427 with Pm21 linkage molecules mark：If when certain pair of primers haynaldia villosa, hard cluster wheat,
T6VS6AL, NAU418 band consistent with being amplified in NAU427, and the band can not expand in China spring, the then primer
Corresponding mark is then to be marked in NAU427 with Pm21 linkage molecules.According to this standard, the present invention develop altogether 25 with
The molecular labeling chain with Pm21 in NAU427：CINAU1692, CINAU1693, CINAU1694, CINAU1695,
CINAU1696, CINAU1697, CINAU1698, CINAU1699, CINAU1700, CINAU1701, CINAU1702,
CINAU1703, CINAU1704, CINAU1705, CINAU1706, CINAU1707, CINAU1708, CINAU1709,
CINAU1710, CINAU1711, CINAU1712, CINAU1713, CINAU1714, CINAU1715, CINAU1716.Primer sequence
Column information is as shown in Table 1.
Embodiment 4. and application of the molecular labeling chain Pm21 in marker assisted selection
Using the minimum fragment translocation line NAU427 of the primer pair marked with Pm21 linkage molecules or its be parent derived from after
Enter performing PCR amplification for the genomic DNA of material, amplified production carries out gel electrophoresis separation, if it is possible to which the molecule is arrived in amplification
Haynaldia villosa particular target band corresponding to mark, then illustrate to contain gene Pm 21 gene in wheat breed to be identified, conversely, then not
Containing the gene.By the exploitation and its utilization pair marked with Pm21 linkage molecules, wheat breeding for disease resistance selection effect is remarkably improved
Rate.
The primer sequence of the molecular labeling chain with Pm21 in the NAU427 of table one
Sequence table
<110>Agricultural University Of Nanjing
Nanjing Xin Maixiu bio tech ltd
<120>Chain molecular labeling and its application in breeding with wheat resistance genes Pm21
<160> 50
<170> SIPOSequenceListing 1.0
<210> 1
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
cgaggggtgc tcccttat 18
<210> 2
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
agctacagaa acacggtaac actgcta 27
<210> 3
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
gtggtgattt tagtttcatg gtgatagtat g 31
<210> 4
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
aaattcacag gctcatggca tgca 24
<210> 5
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
cgctgttaca cttggtgatt gacctttaa 29
<210> 6
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
tggtccttgg caacctcact ttggt 25
<210> 7
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
ttgccaacat ccgagtatcg atgg 24
<210> 8
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 8
agtctacctc acccgttgga cttg 24
<210> 9
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 9
ctctgacatg atatcatgag caattaactc 30
<210> 10
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 10
ggtgatcttc attgcctcca 20
<210> 11
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 11
acttcatgcg ccaatttgga ggatgt 26
<210> 12
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 12
caaggccgcc gaatcctaca at 22
<210> 13
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 13
ctgtacgcgc gagacactgg a 21
<210> 14
<211> 32
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 14
actgttgaat ttgttccact taatacgaga gt 32
<210> 15
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 15
tcttctgacc cgtgcaaatc cagaa 25
<210> 16
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 16
tccagtgtct cgcgcgtaca g 21
<210> 17
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 17
aaattcctac ttttgcgatg tca 23
<210> 18
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 18
aaatcttgcc atgagttgta gggaaaata 29
<210> 19
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 19
aaatcttgcc atgagttgta gggaaaata 29
<210> 20
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 20
caagaggttt ataatatagt gtctgtaga 29
<210> 21
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 21
aagctgatgc ctggtcactc tca 23
<210> 22
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 22
ggctctactt tgtctacaga aaagcc 26
<210> 23
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 23
ggttataggc tgaaatacgt atttcatag 29
<210> 24
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 24
tcatgtattg acgaacggat gt 22
<210> 25
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 25
tctcctattc tgcgtcttcc a 21
<210> 26
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 26
gccagtgaag gcttgatgta 20
<210> 27
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 27
tgttttcttg cagtcgacca tgacaatc 28
<210> 28
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 28
tgttgacatc gcaaaagtag g 21
<210> 29
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 29
gcatccgttc gacgttggct tc 22
<210> 30
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 30
ctctggttgg agggggaggt 20
<210> 31
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 31
gtagcattat agtccgctga attaacc 27
<210> 32
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 32
ccatggcgca tagcataacc attgg 25
<210> 33
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 33
tgttcagagt tcagacagta agcaatgaa 29
<210> 34
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 34
ctgattatgc gattgcctgt 20
<210> 35
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 35
gtttattttt tagttggacc aagttggacc 30
<210> 36
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 36
ttcattgctt actgtctgaa ctctgaaca 29
<210> 37
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 37
catcgttaac caaatcacat cgatctgc 28
<210> 38
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 38
caaggtccaa ttctgctttg ctcc 24
<210> 39
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 39
gatggtcttc ttctttagcg caaatg 26
<210> 40
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 40
tagagataaa tattgtattc cctgagtccc a 31
<210> 41
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 41
gctgaaatga ttgtctgtac actagggt 28
<210> 42
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 42
gtgaactttc tcgagagcac agaaga 26
<210> 43
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 43
gagcgtttag atcactaaag tagattc 27
<210> 44
<211> 29
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 44
gggtttatta tgatatatac tggctatag 29
<210> 45
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 45
tagtgcatcc atacttcaac ttagtc 26
<210> 46
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 46
tcaggatcaa tgtagcagaa gg 22
<210> 47
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 47
gacggaaagg accattttag tgattagc 28
<210> 48
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 48
agatgggagc cataggatca 20
<210> 49
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 49
caacatgttc cggaccctct g 21
<210> 50
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 50
agcgcatctt gaatccttga 20
Claims (6)
1. with chain wheat resistance genes Pm21 molecular labeling, it is characterised in that selected from CINAU1692, CINAU1693,
CINAU1694、CINAU1695、CINAU1696、CINAU1697、CINAU1698、CINAU1699、CINAU1700、
CINAU1701、CINAU1702、CINAU1703、CINAU1704、CINAU1705、CINAU1706、CINAU1707、
CINAU1708、CINAU1709、CINAU1710、CINAU1711、CINAU1712、CINAU1713、CINAU1714、
Any one in CINAU1715 or CINAU1716；Its primer sequence is as shown in table 1：
Table 1
。
2. the primer of the molecular labeling chain with disease-resistant gene Pm21 described in claim 1, it is characterised in that sequence such as SEQ
Shown in ID NO.1-SEQ ID NO.50.
3. application of the primer in Wheat Germplasm Resources of the identification containing Pm21 genes, strain and kind described in claim 2.
4. application of the primer described in claim 2 in wheat breeding for disease resistance.
5. the disease-resistant translocation line NAU427 of the minimum fragment containing resistant gene to wheat powdery mildew Pm21, it is characterised in that be preserved in China
Type Tissue Collection, preservation date 2017.3.17, preserving number CCTCCP201707, preservation address are military for China
Chinese university.
6. the disease-resistant translocation line NAU427 of the minimum fragment containing resistant gene to wheat powdery mildew Pm21 described in claim 5 is in wheat
Application in breeding for disease resistance.
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| CN110923352A (en) * | 2019-12-02 | 2020-03-27 | 河南大学 | KASP marker of wheat powdery mildew resistance gene PmDTM and application thereof |
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| CN1661106A (en) * | 2004-12-30 | 2005-08-31 | 南京农业大学 | Tagged primer chained to Pm21 gene of anti powdery mildew of wheat and application |
| CN101760550A (en) * | 2008-12-19 | 2010-06-30 | 曹淑兰 | Research on location of molecular markers of powdery mildew resistance genes |
| CN102925544A (en) * | 2012-07-03 | 2013-02-13 | 江苏里下河地区农业科学研究所 | Molecular marker for specifically tracking dasypyrumvillosum 6VS chromosome and use method thereof |
| CN104278028A (en) * | 2014-09-16 | 2015-01-14 | 天津师范大学 | Sequence of dasypyrum villosum 6VS DNA permeating into powdery mildew resistant near-isogenic line of wheat and application |
| CN105969778A (en) * | 2016-05-10 | 2016-09-28 | 中国农业大学 | Haynaldia villosa gene NAM-V1 and molecular marker and application thereof |
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| CN109136232A (en) * | 2017-06-19 | 2019-01-04 | 江苏大学 | Haynaldia villosa mildew-resistance gene DvRGA-1, DvRGA-2 and its application |
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2017
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| CN1661106A (en) * | 2004-12-30 | 2005-08-31 | 南京农业大学 | Tagged primer chained to Pm21 gene of anti powdery mildew of wheat and application |
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| CN110923352B (en) * | 2019-12-02 | 2021-07-23 | 河南大学 | KASP marker of wheat powdery mildew resistance gene PmDTM and application thereof |
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