CN109182575A - A kind of method of the anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding - Google Patents
A kind of method of the anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding Download PDFInfo
- Publication number
- CN109182575A CN109182575A CN201811050252.XA CN201811050252A CN109182575A CN 109182575 A CN109182575 A CN 109182575A CN 201811050252 A CN201811050252 A CN 201811050252A CN 109182575 A CN109182575 A CN 109182575A
- Authority
- CN
- China
- Prior art keywords
- corn
- southern rust
- self
- primer
- band
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Botany (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Mycology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
A kind of method that the present invention discloses anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding, belongs to plant molecular technical field.The present invention develops at least one of the molecular labeling, including M0607, M0801, M0904, M1003 and M3402 of 5 pairs with the anti-southern rust close linkage of corn at the disease-resistant section bin10.01 of oriented southern rust.The molecular labeling that the present invention is developed is Indel label, and based on PCR amplified band polymorphism can be detected, at low cost, high reliablity by agarose gel electrophoresis.Method of the invention is a kind of efficient, accurate breeding technique that can obtain anti-southern rust corn genetic stocks in early days；Inbred line of sweet corn after importing disease-resistant section has obtained improving significantly to the resistance of southern rust.The present invention obtains the inbred line of sweet corn of stable highly resistance southern rust in a short time, has significantly speeded up the anti-southern rust improvement process of corn.
Description
Technical field
The invention belongs to plant molecular technical fields, and in particular to 5 pairs with the molecule of the anti-southern rust close linkage of corn
The method of the exploitation of label and the disease-resistant inbred line of sweet corn new material of molecular marking supplementary breeding.
Background technique
Corn southern rust is a kind of gas transmissibility as caused by Puccinia polysora (Pucciniap olysora Undrew)
Disease.The stem of corn after plant is susceptible, leaf, bract surface adhere to the uredium of a large amount of crocus, seriously affect the light of corn
Cooperation is used, and then influences corn yield.The general generation time can cause the underproduction 10%~20%, and the serious time of falling ill can cause to subtract
Produce 50% or more, or even total crop failure.In China, the disease was most found earlier than 1972 in Hainan, was gradually increasing after the nineties as south
The Major Diseases of regional corn, in recent years southern rust diseased region gradually northward extension, 2015 Huang-Huai-Hai summer corn area south
Rust great outburst accounts for the 87.8% of national onset area, causes great production loss [1] to maize production.To China's mesh
Preceding Sweet corn variety find after Resistance Identification most of kind to southern rust show as it is susceptible or in resist, highly resistance kind
Less, when southern rust is caused harm, severity is every to increase 10%, then fresh edible maize seed 100-grain weight will reduce about 0.3g, cultivate resistance
Kind not only can be reduced corn production loss, improve corn harvest quality, while can be effectively controlled bacterium source, in national model
Enclose the interior generation [2,3] for reducing southern rust.
Has the history of many decades to the genetics research of the anti-southern rust of corn.Ullstrup is in No. 10 chromosomes of corn
On just navigated to the disease-resistant gene Rpp9 [4] resistant to microspecies PP.9.Then, for the disease-resistant base of different biological strains
Because being positioned in succession.Nineteen ninety, China breeder draw cenospecies 78599 in addition and are bred as first anti-southern rust for basic material
Corn inbred line neat 319, which shows in the resistant phenotype identification of subsequent many years stablizes, and shows as to southern rust
Highly resistance [5].Chen etc. is navigated to disease-resistant gene RppQ [6] in disease-resistant self-mating system neat 319, and Zhang etc. utilizes disease-resistant self-mating system
W2D and susceptible self-mating system W222 assemble group, navigate to disease-resistant gene RppD [7], and Zhao etc. utilizes a pair of of near isogenic lines
F939 and F349 is navigated to disease-resistant gene Rpp25 [8], and Wu etc. navigates to main effect disease-resistant gene on highly resistance self-mating system SCML205
RppS[9].It is worth noting that, above-mentioned all disease-resistant gene Rpp9, RppQ, RppD, Rpp25, RppS are all located in jade
At No. 10 chromosome bin10.01 of rice, and this region includes the R gene of multiple NBS-LRR types, and scientists speculate herein
There may be a disease-resistant gene rich region, multiple disease-resistant genes form disease-resistant gene cluster, same to different biological strains
When play resistant effect.
It is greatly convenient that the fast development of bioinformatics is provided to breeder's progress molecular breeding, utilizes known base
Because group sequence information can carry out the exploitation and screening of a large amount of molecular labeling.Current anti-southern rust molecular labeling is confined to early stage
A small number of SSR markers, these are marked from resistance locus farther out, and are not suitable for the sweet tea to differ greatly with conventional corn genetic background
Corn.
Bibliography:
[1] Liu Jie, Jiang Yuying, Zeng Juan wait the China .2015 corn southern rust to retransmit in feature and the analysis of causes [J]
State's plant protection guide, 2016,36 (5): 44-47.
[2] Tian Yaojia, Zhao Shouguang, Zhang Jing, wait southern rust on fresh edible maize generation dynamic and its occurrence degree with
Correlation [J] China's Vegetable of yield, 2016,1 (7): 48-51.
[3] Tian Yaojia, Zhao Shouguang, Zhang Hua, at the beginning of waiting the anti-southern rust identification of fresh edible maize cross combination and its genetics of resistance
Analyse [J] agriculture in south journal, 2014,45 (7): 1188-1192.
[4]Ullstrup AJ.Inheritance and linkage of a gene determining
resistance in Maize to an American race of Fucciniapolysora[J]
.Phytopathology,1965,55(4):425-428.
[5] leaf gold ability is bred as China's the first to corn southern rust siberian crabapple neat 319 [J] Scientia Agricultura Sinica, and 2000,
33(4):110-110.
[6]Chen C X,Wang Z L,Yang D E,et al.Molecular tagging and genetic
mapping of the disease resistance gene RppQ to southern corn rust[J]
.Theoretical and Applied Genetics,2004,108(5):945-950.
[7]Zhang Y,Xu L,Zhang D F,et al.Mapping of southern corn rust-
resistant genes in the W2D inbred line of maize(Zea mays L.)[J].Molecular
breeding,2010,25(3):433-439.
[8]Zhao P,Zhang G,Wu X,et al.Fine mapping of RppP25,a southern rust
resistance gene in maize[J].Journal of integrative plant biology,2013,55(5):
462-472.
[9]Wu X,Li N,Zhao P,et al.Geographic and genetic identification of
RppS,a novel locus conferring broad resistance to southern corn rust disease
in China[J].Euphytica,2015,205(1):17-23.
Summary of the invention
In order to overcome the disadvantages and deficiencies of the prior art, the purpose of the present invention is to provide 5 pairs and the anti-southern rust of corn
The molecular labeling of close linkage.
Another object of the present invention is to provide for expanding and the molecular labeling of the anti-southern rust close linkage of corn
Primer pair.
Another object of the present invention is to provide the applications of above-mentioned molecular labeling or primer pair.
A further object of the present invention is to provide a kind of anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding
Method.
The present invention will develop new molecular labeling at bin10.01, and it is beautiful that neat 319 disease-resistant section is imported susceptible sweet tea
Rice self-mating system, improves the resistance of Sweet corn variety.
The purpose of the invention is achieved by the following technical solution:
The present invention provide with the molecular labeling of the anti-southern rust close linkage of corn, the molecular labeling include M0607,
At least one of M0801, M0904, M1003 and M3402.
Expand the primer pair of M0607: forward primer and reverse primer sequences are respectively SEQ ID NO.1 and 2；
Expand the primer pair of M0801: forward primer and reverse primer sequences are respectively SEQ ID NO.3 and 4；
Expand the primer pair of M0904: forward primer and reverse primer sequences are respectively SEQ ID NO.5 and 6；
Expand the primer pair of M1003: forward primer and reverse primer sequences are respectively SEQ ID NO.7 and 8；
Expand the primer pair of M3402: forward primer and reverse primer sequences are respectively SEQ ID NO.9 and 10.
Preferably, the corn is inbred line of sweet corn GQ-1, TZ-1 and conventional corn self-mating system neat 319.
M0607 amplifies the band of 933bp in self-mating system TZ-1, and the item of 695bp is amplified in self-mating system neat 319
Band；M0801 amplifies no band in self-mating system TZ-1, and the band of 639bp is amplified in self-mating system neat 319；M0904 exists
Amplification amplifies the band of 537bp without band in self-mating system neat 319 in GQ-1；M1003 is expanded in self-mating system GQ-1 without item
Band amplifies the band of 1298bp in self-mating system TZ-1, and the band of 919bp is amplified in self-mating system neat 319；M3402 exists
The band that 840bp is amplified in self-mating system GQ-1 amplifies the band of 393bp in self-mating system neat 319.
The improvement of inbred line of sweet corn GQ-1 includes dominant marker M0904, M1003 and codominant marker M3402；Corn
The improvement of self-mating system TZ-1 includes dominant marker M0801 and codominant marker M0607, M1003.
The present invention also provides a kind of for expanding and the primer pair of the molecular labeling of the anti-southern rust close linkage of corn, sequence
Column are following at least a pair of:
Expand the primer pair of M0607: forward primer and reverse primer sequences are respectively SEQ ID NO.1 and 2；
Expand the primer pair of M0801: forward primer and reverse primer sequences are respectively SEQ ID NO.3 and 4；
Expand the primer pair of M0904: forward primer and reverse primer sequences are respectively SEQ ID NO.5 and 6；
Expand the primer pair of M1003: forward primer and reverse primer sequences are respectively SEQ ID NO.7 and 8；
Expand the primer pair of M3402: forward primer and reverse primer sequences are respectively SEQ ID NO.9 and 10.
Preferably, the corn is inbred line of sweet corn GQ-1, TZ-1 and conventional corn self-mating system neat 319.
The present invention provides a kind of application of above-mentioned molecular labeling or primer pair in assisted Selection improvement corn, specifically
, the application in the anti-southern rust inbred line of sweet corn of breeding.
The present invention provides a kind of method of anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding, including following step
It is rapid:
1) high sense southern rust corn Inbred Lines GQ-1, TZ-1 are neat with highly resistance southern rust Inbred Lines respectively
319 hybridization carry out four backcrossings as recurrent parent using corn Inbred Lines and obtain BC4F1Backcross population；
2) BC is extracted4F1Backcross population DNA carries out PCR amplification to backcross population DNA according to the combination of above-mentioned molecular labeling, leads to
Agarose gel electrophoresis detection PCR product stripe size is crossed, the positive plant for having neat 319 disease-resistant chromosome segments is obtained；
3) selfing is carried out to the positive plant in step 2) and obtains BC4F2Group；
4) BC is extracted4F2Group DNA carries out PCR amplification to DNA according to the codominant marker in the combination of above-mentioned molecular labeling,
Positive plant after being improved, with homozygous disease-resistant chromosome segment；
5) it is planted with the plant after improvement in morbidity field before improveing, it is for statistical analysis to disease index, it obtains respectively
Obtain anti-southern rust inbred line of sweet corn GQ-1R, TZ-1R.
The present invention has the following advantages and effects with respect to the prior art:
(1) molecular labeling that the present invention is developed is Indel label, and based on PCR amplified band polymorphism can pass through agar
Sugared gel electrophoresis is detected, at low cost, high reliablity.
(2) method of the invention is that one kind can obtain the efficient, accurate of anti-southern rust corn genetic stocks in early days
Breeding technique.
(3) present invention develops 5 pairs at the disease-resistant section bin10.01 of oriented southern rust and closely connects with disease-resistant section
The molecular labeling of lock, and by these label screening Advanced backcross groups, by the disease-resistant section of conventional corn self-mating system neat 319
Inbred line of sweet corn GQ-1, the TZ-1 for importing high sense southern rust, obtain self-mating system GQ-1R, TZ-1R of highly resistance southern rust.
The result shows that the inbred line of sweet corn imported after disease-resistant section has obtained improving to the resistance of southern rust significantly.The present invention
The inbred line of sweet corn for obtaining stable highly resistance southern rust in a short time has significantly speeded up the anti-southern rust of corn
Improvement process.
Detailed description of the invention
Fig. 1 is testing result of the difference molecular labeling between two parents；Wherein, M DL2000Marker.
Fig. 2 is the positive plant that molecular marker screening has imported disease-resistant chromosome segment；Wherein, it is shown altogether using molecular labeling
Property label M3402 detection using GQ-1 as the BC of recurrent parent4F1The agarose gel electrophoresis results of backcross population: swimming lane 1~23 is
BC4F1Backcross population；Swimming lane 24 is GQ-1；Swimming lane 25 is neat 319；Swimming lane M is DL2000Marker.
Fig. 3 is the positive plant that molecular marker screening has imported disease-resistant chromosome segment；Wherein, it is shown altogether using molecular labeling
Property label M1003 detection using TZ-1 as the BC of recurrent parent4F1The agarose gel electrophoresis results of backcross population: swimming lane 1~20 is
BC4F1Backcross population；Swimming lane 21 is TZ-1；Swimming lane 22 is neat 319；Swimming lane M is DL2000Marker.
Fig. 4 is disease index of the inbred line of sweet corn in morbidity field of improvement front and back；Wherein, * *, t-test, p <
0.01。
Fig. 5 is the field phenotype of the inbred line of sweet corn of improvement front and back.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
The test method of specific experiment condition is not specified in the following example, usually according to conventional laboratory conditions or according to system
Make experiment condition proposed by factory.
Following biomaterial involved in embodiment:
Inbred line of sweet corn GQ-1 is disclosed as the parent of Guangdong sweet tea No. 27 (state examines jade 20170038), open network address:
http://www.moa.gov.cn/govpublic/nybzzj1/201707/t20170712_5745606.htm。
Miscellaneous -1 (the abbreviation TZ-1) in inbred line of sweet corn field is that the parent of Guangdong sweet tea 27 (state examines jade 20170038) discloses, open
Network address: http://www.moa.gov.cn/govpublic/nybzzj1/201707/t20170712_574560 6.htm.
Corn inbred line neat 319 " is bred as China's the first to the agriculture of neat 319 [J] China of corn southern rust siberian crabapple in document
Industry science, 2000,33 (4): discloses in 110-110. ".
Embodiment 1
1. the DNA sequence dna at the website Maizesequence downloading corn bin10.01, design primer expand corn
The genomic DNA of self-mating system GQ-1, field miscellaneous -1 (abbreviation TZ-1) and conventional corn self-mating system neat 319.PCR product is subjected to agar
The primer that amplified product band has differences between inbred line of sweet corn and neat 319 is determined as molecular labeling by sugared gel electrophoresis.
By obtaining 5 pairs of molecules that can be used for the anti-southern rust assisted selection of corn altogether after carrying out gel electrophoresis to PCR product
Label.The primer sequence of amplifier molecule label is as shown in table 1, and type is as shown in table 2.
The primer sequence of 1 amplifier molecule of table label
The type of the anti-southern rust molecular labeling of table 2
| M0607 | M0801 | M0904 | M1003 | M3402 | |
| GQ-1/ neat 319 | - | - | 1 | 1 | 2 |
| TZ-1/ neat 319 | 2 | 1 | - | 2 | - |
Note: " 1 ": dominant marker；" 2 ": codominant marker；"-": indifference.
Testing result of the difference molecular labeling between two parents, as shown in Figure 1.As a result it indicates: inbred line of sweet corn GQ-
1 improvement includes dominant marker M0904, M1003 and codominant marker M3402；The improvement of inbred line of sweet corn TZ-1 includes aobvious
Property label M0801 and codominant marker M0607, M1003.
2. inbred line of sweet corn GQ-1, TZ-1 are hybridized with conventional corn self-mating system neat 319 respectively, then with sweet tea jade
Rice self-mating system is that recurrent parent carries out mostly obtaining BC for backcrossing4F1Backcross population.
3. extracting BC4F1Backcross population DNA carries out PCR amplification to DNA with the primer sequence that amplifier molecule in table 1 marks,
Agarose gel electrophoresis is carried out to PCR product, has imported purpose section (neat 319 disease-resistant chromosomal region by running the determination of adhesive tape band
Section) positive plant.It is detected using codominant marker M3402 using GQ-1 as the BC of recurrent parent4F1The agarose of backcross population is solidifying
Gel electrophoresis is as a result, as shown in Fig. 2, plant 1,2,3,4,12,16,18, No. 20 identical as donor parents neat 319 (No. 25), to lead
The positive plant of disease-resistant chromosome segment is entered.It is detected using codominant marker M1003 using TZ-1 as the BC of recurrent parent4F1It returns
The agarose gel electrophoresis results of group are handed over, as shown in Figure 3.As a result it indicates, plant 1,2,3,4,8,9,10,11,12,13,15,
16,18,19, No. 20 bands are identical as donor parents neat 319 (No. 22), plant to have imported the positive of disease-resistant chromosome segment
Strain.
4. the positive plant in step 3 is selfed, BC is obtained4F2Group；With Markers for Detection self progeny group
Body screens the disease-resistant self-mating system of homozygous dominant, the inbred line of sweet corn of the anti-southern rust after being improved.
5. by the plant before improvement and after improvement in morbidity field plantation, field test southern rust phenotype calculates the state of an illness
Whether index, the self-mating system after judging improvement improve the resistance of southern rust.Anti- southern rust corn is finally obtained respectively
Self-mating system GQ-1R, TZ-1R.
Single plant investigation is carried out to leaf, 3 leaf of fringe inferior lobe on the fringe position leaf of corn, fringe in corn milk stage, leaf is accounted for according to scab
The percentage of the gross area of piece is classified.Grade scale is as shown in table 3.
3 severity Scaling standard of table
| State of an illness rank | Symptom description |
| 1 | Disease-free spot on blade, or only whether there is or not the hypersensitive necrosis reactions of sorus |
| 3 | There is a small amount of sorus on blade, accounts for leaf area less than 25% |
| 5 | There is middle amount sorus on blade, accounts for leaf area 26%~50% |
| 7 | There are a large amount of sorus on blade, accounts for leaf area 51%~75% |
| 9 | The a large amount of sorus in blade upstream, account for leaf area 76%~100%, blade is withered |
The calculation method of disease index is as follows:
Disease index=∑ (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × superlative degree value) × 100.
Disease index of the inbred line of sweet corn in morbidity field of front and back is improved, as shown in Figure 4.As a result it indicates, before improvement,
The disease index in the field GQ-1 is 0.71, for high sense material.The disease index of GQ-1R is 0.15 after improvement, shows as highly resistance.Change
The disease index of good preceding TZ-1 is 0.73, and height sense southern rust, the disease index of TZ-1R is 0.14 after improvement, shows as highly resistance.
The field phenotype of the inbred line of sweet corn of front and back is improved, as shown in Figure 5.As a result it indicates, does not improve to improvement and material
The incidence of material milk stage is observed, and is photographed to record.As shown in figure 5, the fringe position leaf of GQ-1 and TZ-1 before improvement
There is a large amount of pathogen spore on blade, pathogen spore is linked to be piece, causes serious harm to blade.GQ-1R after improvement with
The blade surface disease-free spot of TZ-1 or fragmentary hypersensitive necrosis spot are highly resistance phenotype.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Sequence table
<110>Crop Institute, Guangdong Academy of Agricultural Sciences
<120>method of the anti-southern rust inbred line of sweet corn of a kind of molecular marking supplementary breeding
<160> 10
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M0607 forward primer
<400> 1
gatttgcatg ggaccaggac 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M0607 reverse primer
<400> 2
tgttgcgacg cttccatatg 20
<210> 3
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M0801 forward primer
<400> 3
agctcgatca atccagggag 20
<210> 4
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M0801 reverse primer
<400> 4
gataccgtgg ttagaggcga 20
<210> 5
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M0904 forward primer
<400> 5
cacaagcagg atggtagcac 20
<210> 6
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M0904 reverse primer
<400> 6
tcctccagaa tccagatgca 20
<210> 7
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M1003 forward primer
<400> 7
tgcacgccat cacaaatcaa 20
<210> 8
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M1003 reverse primer
<400> 8
ccttcatgac gcttgctctc 20
<210> 9
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M3402 forward primer
<400> 9
ctgctatgac aagtgccgac 20
<210> 10
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<220>
<223>M3402 reverse primer
<400> 10
cctctgccta atgaaacgcc 20
Claims (9)
1. with the molecular labeling of the anti-southern rust close linkage of corn, it is characterised in that: the molecular labeling include M0607,
At least one of M0801, M0904, M1003 and M3402；
Expand the primer pair of M0607: forward primer and reverse primer sequences are respectively SEQ ID NO.1 and 2；
Expand the primer pair of M0801: forward primer and reverse primer sequences are respectively SEQ ID NO.3 and 4；
Expand the primer pair of M0904: forward primer and reverse primer sequences are respectively SEQ ID NO.5 and 6；
Expand the primer pair of M1003: forward primer and reverse primer sequences are respectively SEQ ID NO.7 and 8；
Expand the primer pair of M3402: forward primer and reverse primer sequences are respectively SEQ ID NO.9 and 10.
2. the according to claim 1 and anti-southern rust close linkage of corn molecular labeling, it is characterised in that:
The corn is inbred line of sweet corn GQ-1, TZ-1 and conventional corn self-mating system neat 319.
3. the according to claim 2 and anti-southern rust close linkage of corn molecular labeling, it is characterised in that:
M0607 amplifies the band of 933bp in self-mating system TZ-1, and the band of 695bp is amplified in self-mating system neat 319；
M0801 amplifies no band in self-mating system TZ-1, and the band of 639bp is amplified in self-mating system neat 319；M0904 is in GQ-1
Middle amplification amplifies the band of 537bp without band in self-mating system neat 319；M1003 is expanded in self-mating system GQ-1 without band,
The band that 1298bp is amplified in self-mating system TZ-1 amplifies the band of 919bp in self-mating system neat 319；M3402 is being selfed
It is the band for amplifying 840bp in GQ-1, the band of 393bp is amplified in self-mating system neat 319.
4. the according to claim 2 or 3 and anti-southern rust close linkage of corn molecular labeling, it is characterised in that:
The improvement of inbred line of sweet corn GQ-1 includes dominant marker M0904, M1003 and codominant marker M3402；Corn selfing
The improvement for being TZ-1 includes dominant marker M0801 and codominant marker M0607, M1003.
5. a kind of for expanding and the primer pair of the molecular labeling of the anti-southern rust close linkage of corn, it is characterised in that: described
The sequence of primer pair is at least a pair of as follows:
Expand the primer pair of M0607: forward primer and reverse primer sequences are respectively SEQ ID NO.1 and 2；
Expand the primer pair of M0801: forward primer and reverse primer sequences are respectively SEQ ID NO.3 and 4；
Expand the primer pair of M0904: forward primer and reverse primer sequences are respectively SEQ ID NO.5 and 6；
Expand the primer pair of M1003: forward primer and reverse primer sequences are respectively SEQ ID NO.7 and 8；
Expand the primer pair of M3402: forward primer and reverse primer sequences are respectively SEQ ID NO.9 and 10.
6. it is according to claim 5 for expand and the primer pair of the molecular labeling of the anti-southern rust close linkage of corn,
It is characterized by:
The corn is inbred line of sweet corn GQ-1, TZ-1 and conventional corn self-mating system neat 319.
7. primer pair described in the described in any item molecular labelings of Claims 1 to 4 or claim 5~6 changes in assisted Selection
Application in good corn.
8. application according to claim 7, it is characterised in that:
The application of the molecular labeling or the primer pair in the anti-southern rust inbred line of sweet corn of breeding.
9. a kind of method of the anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding, it is characterised in that including following step
It is rapid:
1) high sense southern rust corn Inbred Lines GQ-1, TZ-1 respectively with highly resistance southern rust Inbred Lines neat 319
Hybridization carries out four backcrossings as recurrent parent using corn Inbred Lines and obtains BC4F1Backcross population；
2) BC is extracted4F1Backcross population DNA, molecular labeling combination according to any one of claims 1 to 4 is to backcross population
DNA carries out PCR amplification, detects PCR product stripe size by agarose gel electrophoresis, obtains and have neat 319 disease-resistant chromosomes
The positive plant of section；
3) selfing is carried out to the positive plant in step 2) and obtains BC4F2Group；
4) BC is extracted4F2Group DNA, the codominant marker in molecular labeling according to any one of claims 1 to 4 combination
PCR amplification, positive plant after being improved, with homozygous disease-resistant chromosome segment are carried out to DNA；
5) it is planted with the plant after improvement in morbidity field before improveing, it is for statistical analysis to disease index, resisted respectively
Southern rust inbred line of sweet corn GQ-1R, TZ-1R.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811050252.XA CN109182575B (en) | 2018-09-10 | 2018-09-10 | Method for molecular marker-assisted breeding of southern rust resistant sweet corn inbred line |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811050252.XA CN109182575B (en) | 2018-09-10 | 2018-09-10 | Method for molecular marker-assisted breeding of southern rust resistant sweet corn inbred line |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109182575A true CN109182575A (en) | 2019-01-11 |
| CN109182575B CN109182575B (en) | 2021-07-06 |
Family
ID=64915757
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811050252.XA Active CN109182575B (en) | 2018-09-10 | 2018-09-10 | Method for molecular marker-assisted breeding of southern rust resistant sweet corn inbred line |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109182575B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112216343A (en) * | 2020-10-28 | 2021-01-12 | 遵义市农业科学研究院 | Screening method of tomato spotted wilt virus resistant hot pepper plants |
| CN113631722A (en) * | 2019-03-11 | 2021-11-09 | 先锋国际良种公司 | Methods for identifying, selecting and producing southern corn rust resistant crops |
| CN113897352A (en) * | 2020-06-22 | 2022-01-07 | 北京市农林科学院 | Corn southern rust resistance gene close linkage marker and application thereof |
| CN116941525A (en) * | 2023-08-22 | 2023-10-27 | 河南农业大学 | Improved method for improving southern rust resistance of corn hybrid by increasing resistance due to multiple factors |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105724241A (en) * | 2016-03-04 | 2016-07-06 | 湖北康农种业股份有限公司 | Method for breeding maize inbred line KNL1088 resistant to frogeye leaf spot and southern rust |
| CN105861647A (en) * | 2016-01-26 | 2016-08-17 | 河南农业大学 | Molecular marker combination realizing close linkage with southern corn rust gene and applications of molecular marker combination |
| CN106282394A (en) * | 2016-10-28 | 2017-01-04 | 中玉金标记（北京）生物技术股份有限公司 | The method of high throughput testing Semen Maydis south rust resistance gene type and test kit thereof |
| US20170367280A1 (en) * | 2016-05-27 | 2017-12-28 | Kws Saat Se | Use of argonaute endonucleases for eukaryotic genome engineering |
-
2018
- 2018-09-10 CN CN201811050252.XA patent/CN109182575B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105861647A (en) * | 2016-01-26 | 2016-08-17 | 河南农业大学 | Molecular marker combination realizing close linkage with southern corn rust gene and applications of molecular marker combination |
| CN105724241A (en) * | 2016-03-04 | 2016-07-06 | 湖北康农种业股份有限公司 | Method for breeding maize inbred line KNL1088 resistant to frogeye leaf spot and southern rust |
| US20170367280A1 (en) * | 2016-05-27 | 2017-12-28 | Kws Saat Se | Use of argonaute endonucleases for eukaryotic genome engineering |
| CN106282394A (en) * | 2016-10-28 | 2017-01-04 | 中玉金标记（北京）生物技术股份有限公司 | The method of high throughput testing Semen Maydis south rust resistance gene type and test kit thereof |
Non-Patent Citations (3)
| Title |
|---|
| C X CHEN等: "Molecular tagging and genetic mapping of the disease resistance gene RppQ to southern corn rust", 《THEOR APPL GENET》 * |
| CHUNJIANG ZHOU等: "Characterization and fine mapping of RppQ, a resistance gene to southern corn rust in maize", 《MOL GENET GENOMICS》 * |
| 姚国旗等: "玉米自交系CML470 抗南方锈病基因的定位", 《植物遗传资源学报》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113631722A (en) * | 2019-03-11 | 2021-11-09 | 先锋国际良种公司 | Methods for identifying, selecting and producing southern corn rust resistant crops |
| CN113631722B (en) * | 2019-03-11 | 2024-05-28 | 先锋国际良种公司 | Methods for identifying, selecting and producing southern corn rust resistant crops |
| CN113897352A (en) * | 2020-06-22 | 2022-01-07 | 北京市农林科学院 | Corn southern rust resistance gene close linkage marker and application thereof |
| CN113897352B (en) * | 2020-06-22 | 2023-06-23 | 北京市农林科学院 | Closely linked marker of southern rust resistance gene of corn and application thereof |
| CN112216343A (en) * | 2020-10-28 | 2021-01-12 | 遵义市农业科学研究院 | Screening method of tomato spotted wilt virus resistant hot pepper plants |
| CN112216343B (en) * | 2020-10-28 | 2023-08-15 | 遵义市农业科学研究院 | Screening method of tomato spotted wilt virus resistant pepper plants |
| CN116941525A (en) * | 2023-08-22 | 2023-10-27 | 河南农业大学 | Improved method for improving southern rust resistance of corn hybrid by increasing resistance due to multiple factors |
| CN116941525B (en) * | 2023-08-22 | 2024-05-28 | 河南农业大学 | Improved method for improving southern rust resistance of corn hybrid by increasing resistance due to multiple factors |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109182575B (en) | 2021-07-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sandhu et al. | Positive interactions of major-effect QTLs with genetic background that enhances rice yield under drought | |
| Rajpurohit et al. | Pyramiding of two bacterial blight resistance and a semidwarfing gene in Type 3 Basmati using marker-assisted selection | |
| Asnaghi et al. | Targeted mapping of a sugarcane rust resistance gene (Bru 1) using bulked segregant analysis and AFLP markers | |
| Kordrostami et al. | Molecular markers in plants: concepts and applications | |
| Zhang et al. | Genetic and correlation analysis of silique-traits in Brassica napus L. by quantitative trait locus mapping | |
| Zhao et al. | Identification of QTLs and meta‐QTLs for seven agronomic traits in multiple maize populations under well‐watered and water‐stressed conditions | |
| CN109182575A (en) | A kind of method of the anti-southern rust inbred line of sweet corn of molecular marking supplementary breeding | |
| EP2735619A2 (en) | Methods and compositions for breeding for preferred traits associated with Goss' Wilt resistance in plants | |
| Wu et al. | Analysis on genetic diversification and heterosis in autotetraploid rice | |
| Tessmann et al. | GWAS for Fusarium head blight traits in a soft red winter wheat mapping panel | |
| Yang et al. | A strategy to develop molecular markers applicable to a wide range of crosses for marker assisted selection in plant breeding: a case study on anthracnose disease resistance in lupin (Lupinus angustifolius L.) | |
| Dong et al. | Marker-assisted selection and evaluation of high oil in vivo haploid inducers in maize | |
| Ezenwaka et al. | Genome‐wide association study of resistance to cassava green mite pest and related traits in cassava | |
| CN107435066B (en) | Rice stigma exposure rate main effect QTL and positioning method and application thereof | |
| Fondevilla et al. | Identification of quantitative trait loci involved in resistance to Pseudomonas syringae pv. syringae in pea (Pisum sativum L.) | |
| Yadav et al. | Morpho-physiological characterization of bread wheat genotypes and their molecular validation for rust resistance genes Sr2, Sr31 and Lr24 | |
| Ma et al. | Genetic analysis and molecular mapping of a stripe rust resistance gene derived from Psathynrostachys huashanica Keng in wheat line H9014-121-5-5-9 | |
| Şahin-Çevik et al. | Quantitative trait loci analysis of morphological traits in Citrus | |
| Micic et al. | QTL mapping of resistance to Sclerotinia midstalk rot in RIL of sunflower population NDBLOS sel× CM625 | |
| Kolmer et al. | Endemic and panglobal genetic groups, and divergence of host-associated forms in worldwide collections of the wheat leaf rust fungus Puccinia triticina as determined by genotyping by sequencing | |
| Ohlson et al. | Identification and mapping of late blight resistance quantitative trait loci in tomato accession PI 163245 | |
| Jangra et al. | Introgression of terminal drought stress tolerance in advance lines of popular pearl millet hybrid through molecular breeding | |
| Ramalingam et al. | Improved host-plant resistance to Phytophthora rot and powdery mildew in soybean (Glycine max (L.) Merr.) | |
| Mahé et al. | Development of sequence characterized DNA markers linked to leaf rust (Hemileia vastatrix) resistance in coffee (Coffea arabica L.) | |
| Zhou et al. | Molecular mapping of a stripe rust resistance gene in wheat cultivar Wuhan 2 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221130 Address after: 518000 Xinghe WORLDA801-02, No.1 Yabao Road, Nankeng Community, Bantian Street, Longgang District, Shenzhen, Guangdong Patentee after: Shenzhen a corn Technology Co.,Ltd. Address before: No.18, Jinying West 2nd Street, Jinying Road, Tianhe District, Guangzhou, Guangdong 510640 Patentee before: CROP Research Institute GUANGDONG ACADEMY OF AGRICULTURAL SCIENCES |