CN1246477C - Molecule marker method of wheat Nanda 2419 scab resistant major gene locus - Google Patents

Molecule marker method of wheat Nanda 2419 scab resistant major gene locus Download PDF

Info

Publication number
CN1246477C
CN1246477C CN 03131961 CN03131961A CN1246477C CN 1246477 C CN1246477 C CN 1246477C CN 03131961 CN03131961 CN 03131961 CN 03131961 A CN03131961 A CN 03131961A CN 1246477 C CN1246477 C CN 1246477C
Authority
CN
China
Prior art keywords
wheat
major gene
gene loci
nanjing university
scab resistance
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.)
Expired - Fee Related
Application number
CN 03131961
Other languages
Chinese (zh)
Other versions
CN1462807A (en
Inventor
马正强
林峰
孔忠新
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Agricultural University
Original Assignee
Nanjing Agricultural University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Nanjing Agricultural University filed Critical Nanjing Agricultural University
Priority to CN 03131961 priority Critical patent/CN1246477C/en
Publication of CN1462807A publication Critical patent/CN1462807A/en
Application granted granted Critical
Publication of CN1246477C publication Critical patent/CN1246477C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Landscapes

  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The present invention provides a major gene site and a molecular marker for resisting the spread of a gibberellic disease of wheat 'Nanda 2419' and belongs to the field of molecular genetics. Genetic linkage analysis is carried out to the gene types, the number of affected spikelet, and the length of an affected section in each line of recombined selfing lines obtained through the hybridization of 'wangshuibai' (female) and 'nanda 2419' (male) which are varieties of wheat so as to obtain the major gene sites QFhS. nau-4b and QFhs. nau-6d1 for resisting the spread of the gibberellic disease of wheat 'Nanda 2419'. By detecting whether the 'nanda 2419' and derived varieties (series) thereof contain the major gene sites or not, the level of the resistance to the gibberellic disease can be predicted, and the selection efficiency of wheat resisting the gibberellic disease is greatly improved.

Description

The molecule marking method of wheat Nanjing University 2419 anti gibberellic diseases expansion major gene loci
One, technical field
The invention provides the molecule marking method of wheat Nanjing University 2419 anti gibberellic diseases expansion major gene loci, belong to the molecular genetics field, be exclusively used in the seed selection of wheat scab disease-resistant variety and the utilization of germ plasm resource.
Two, technical background
Wheat is the important food crop of China.By the wheat scab that gibberella causes, not only cause the serious underproduction of wheat, reduce grain quality, and the toxin deoxynivalenol that this germ produces also influences people, animal health, caused very big loss to Wheat Production.In China, the harm that suffers head blight above 1/4th wheat belt is arranged, and just once be very popular every 3 ~ 5 years, only head blight generation area in 2002 promptly reaches 30,000,000 mu times.Therefore, research prevents and treats wheat scab and has also just become a urgent task on the current Wheat Production.Cultivating disease-resistant variety is to prevent and treat the safest, effective of wheat scab harm and save one of measure of cost.But traditional breeding way is wasted time and energy, and because the head blight phenotypic evaluation is very difficult, makes the wheat anti gibberellic disease breeding process slow unusually.Identify that by the location disease-resistant major gene loci comes assistant breeding effectively to address this problem.
Genetic research shows that wheat scab is controlled by a few major gene loci mainly, and its disease-resistant major gene loci position on karyomit(e) of different resistant materials is inconsistent, and be divided into anti-infect and resist expand two types.But the polymerization by the disease-resistant major gene loci of difference or introduce new anti-source and can select the good kind of resistance.
At present in the utilization of the research of scab resistance hereditary basis and breeding for disease resistance, focus mostly in Soviet Union wheat No. 3 and derive and be.Nanjing University 2419 is choosing systems of Italian wheat breed Mentana, has yielding ability preferably, and economical character is good, is one of backbone parent of China's wheat breeding.And the parent " Funo " of Mentana and Soviet Union three has the common ancestors, so may exist in the Nanjing University 2419 and scab resistance mechanism like the Su Sanxiang.
Three, summary of the invention
Technical problem
The objective of the invention is: the molecule marking method that wheat Nanjing University 2419 anti gibberellic diseases expansion major gene loci is provided, by detecting and the chain molecule marker of these anti-expansion major gene locis, can predict the scab resistance of wheat plant, accelerate the selection progress of wheat-resistance to scab.
Technical scheme
The molecule marking method of wheat Nanjing University 2419 anti gibberellic diseases expansion major gene loci is characterized in that:
Use labeled primer Xgwm107,
Left end primer sequence ATTAATACCTGAGGGAGGTGC
Right-hand member primer sequence GGTCTCAGGAGCAAGAACAC
Amplification wheat anti gibberellic disease kind or breeding material DNA, if can amplify the amplified fragments of 188bp, then indicate the existence of the 2419 anti gibberellic diseases expansion major gene loci Qfhs.nau-4b of wheat Nanjing University, wherein this major gene loci is positioned at the 4B karyomit(e) of wheat Nanjing University 2419, utilize Data desk v.5.0 software record with the incoherent probability P value of scab resistance be 0.0004, to the contribution rate 17.9% of scab resistance;
Perhaps use labeled primer Xgwm469,
Left end primer sequence CAACTCAGTGCTCACACAACG
Right-hand member primer sequence CGATAACCACTCATCCACACC
Amplification wheat anti gibberellic disease kind or breeding material DNA, if can amplify the amplified fragments of 172bp, then indicate the existence of the 2419 anti gibberellic diseases expansion major gene loci Qfhs.nau-6d1 of wheat Nanjing University, wherein this major gene loci is positioned at the 6D karyomit(e) of wheat Nanjing University 2419, utilize Data desk v.5.0 software record with the incoherent probability P value of scab resistance be 0.0089, to the contribution rate 6.1% of scab resistance.
The process of screening above-mentioned labeled primer is as follows:
(1) wheat breed Wangshuibai (♀) and Nanjing University 2419 (♂) hybridize and obtain hybrid F1, and the F1 selfing produces F2, adopts simple grain transmission method (SSD) to obtain the F6 RIL in generation then;
(2) extract the DNA that each strain of recombinant inbred lines is with the SDS method, adopt simple repeated sequence mark SSR that two parents are carried out the polymorphism screening, PCR carries out on PE9600 amplification instrument, amplified production carries out electrophoretic analysis on 8% (g/ml) polyacrylamide gel, there is polymorphic primer in recombinant inbred lines, to analyze between the parent, the pcr amplification program is the same, obtains colony's genotype data;
(3) according to chain exchange rule, utilize colony's genotype data to make up the genetic map of wheat, used software is Mapmaker 2.0, minimum LOD value is made as 3, obtains linkage map;
(4) blooming stage carries out the scab resistance evaluation to each family of RIL, and leading indicator is disease spikelet number and sick joint length
(5) utilize Data desk v.5.0 software carry out linkage analysis to colony's genotype data of each molecule marker with disease tassel yield that the scab resistance of its corresponding each family is identified, One-way ANOVA records and the incoherent probability P value of scab resistance, the molecule marker of P＜0.05 is promptly chain with a major gene loci, the position of anti gibberellic disease expansion major gene loci is determined by the chromosome position of molecule marker: find the Xgwm107 mark of P=0.0004 and the Xgwm469 mark and anti-expansion major gene loci close linkage of P=0.0089 in Nanjing University 2419, Xgwm107 and Xgwm469 are the labeled primer of 2419 anti-expansion major gene loci QFhs.nau-4b of labeling wheat Nanjing University and QFhs.nau-6d1.
The molecule marking method of beneficial effect wheat provided by the present invention Nanjing University 2419 anti gibberellic diseases expansion major gene loci has the following advantages:
(1) obtain first in the world to have located the major gene loci that 2 anti gibberellic diseases in the wheat breed Nanjing University 2419 are expanded by molecule marker of the present invention, they are soluble 39.6% scab resistance jointly.The wheat cdna group is huge, is 40 times of rice genome size, and the accurate location work of wheat anti gibberellic disease major gene loci is occupy the same domain prostatitis;
(2) by the localized major gene loci locality specific of molecule marker mark of the present invention, it is convenient to identify.By detecting these and the chain molecule marker of anti-expansion major gene loci, promptly can predict the scab resistance of wheat plant, the genotype detection that is used for wheat breed or strain, judging whether this kind or strain have scab resistance, and then rapid screening disease-resistant variety or strain are used for wheat breeding.Major gene loci easy to detect fast, not affected by environment;
(3) the assistant breeding select target is clear and definite, saves cost.In traditional breeding way, at first to collect parent and Cultivar and carry out a series of hybridization, and will carry out individual plant to scab resistance and select with disease-resistant gene.Wheat scab is carried out phenotypic evaluation will wait until blooming stage, be subjected to bigger environmental influence, the result reliability of phenotypic evaluation is low.Therefore breeding for disease resistance is not only time-consuming, and difficulty is big, the cost height.By detecting the anti gibberellic disease major gene loci, can just identify the individual plant of high anti gibberellic disease in seedling stage, eliminate other plant, not only save production cost but also improve the efficiency of selection of wheat-resistance to scab greatly.
Four, embodiment
Of the present invention being described in detail as follows:
Studies show that wheat scab resistance is controlled by the minority major gene loci mainly.U.S. Anderson laboratory finds that the scab resistance of No. 3/Stoa of four major gene locis and Soviet Union wheat colony is significantly relevant, lays respectively at 2AL, 3BS, 4BS and 6BS and goes up (Anderson, 1998; Waldron, 1999; Anderson, 2000,2001).People such as U.S. Bai obtain two the scab resistance sites relevant with the RAPD mark, and Zhou integrated use SSR, AFLP in 2002 and aneuploid technology are positioned at this major gene loci that to lack be that 3BS-8 does not hold a last length to be about the chromosomal region of 8cM.Japan Ban infers that the resistance main effect gene locus of Soviet Union wheat 3 may be positioned on the 5AL.Austrian Buerstmayr (2000,2001) finds that three genome areas and the anti-extendability significant correlation of head blight are arranged, and is positioned on 3BS, 5A, the 1B.Molecule marker by wheat of the present invention Nanjing University 2419 anti gibberellic diseases expansion major gene loci is found, 2 anti-expansion major gene locis that exist in the 4B of wheat breed Nanjing University 2419 and the 6D karyomit(e), these major gene locis can be used to instruct the seed selection work of anti gibberellic disease kind, screen with chain with it molecule marker enantiopathy kind, make different disease-resistant major gene loci rapid polymerizations in same plant, thereby improve breeding efficiency greatly.
Materials and methods:
(1) structure and the phenotypic evaluation of Nanjing University's 2419 RILs:
(1) choosing to China's Jiangsu local variety Wangshuibai (♀) and Italian wheat breed Mentana is that acquisition F1 is hybridized in Nanjing University 2419 (♂), the F1 selfing produces F2,136 optional plantations of F2 individual plant, the method that adopts simple grain to pass is bred F6 generation, obtain RIL, comprise 136 familys;
(2) RIL is planted in Agricultural University Of Nanjing in experimental plot, the greenhouse and academy of agricultural sciences, Jiangsu Province in the school.Blooming stage carries out scab resistance to each family to be identified.Adopt single flower instillation, promptly choose the small ear of just having bloomed in wheat head middle part, instiling contains mixed bacteria liquid 20 microlitres of F4, F15, several strong gibberella spores that cause a disease of F17, F34, and atomizing was preserved moisture 3 days then.Inoculate after 21 days, measure state of an illness indexs such as disease spikelet number and sick joint length.
(2) molecular marker analysis of recombinant inbred lines
(1) extracting each strain of recombinant inbred lines with the SDS method is DNA;
(2) at first Wangshuibai and 2419 parents' of Nanjing University dna polymorphism is carried out initial analysis with 976 pairs of SSR primers.The PCR reaction volume is 25 microlitres, 10 * buffer, 2.5 microlitres wherein, 25mM MgCl 21.5 microlitre, 2.5mM dNTPs 2 microlitres, Taq enzyme (5 units/microlitre) 0.2 microlitre, template DNA 20 nanograms add water to 25 microlitres.After the SSR reaction system is 94 ℃ of pre-sex change 3min of DNA, 94 ℃ of sex change 1min, 60 ℃ of annealing 1min, 72 ℃ are extended exhibition 2min, circulate 35 times, and last 72 ℃ are extended 10min.In the enterprising performing PCR amplification of PE 9600 amplification instrument, amplified production carries out electrophoretic separation on 8% non-denaturing polyacrylamide gel (containing 7.6 gram acrylamides and 0.4 gram methylene diacrylamide in the 100ml polyacrylamide solution), on ultraviolet transilluminator, take a picture then, the record result, there is polymorphic primer in recombinant inbred lines, to analyze between the parent, obtains colony's genotype data;
(3) according to chain exchange rule, utilize colony's genotype data to make up the genetic map of wheat, used software is Mapmaker 2.0, minimum LOD value is made as 3, obtains linkage map;
(4) utilize Data desk v.5.0 software carry out linkage analysis, the location major gene loci to colony's genotype data of each molecule marker with sick spikelet number and sick joint length that the scab resistance of its corresponding each family is identified.
(3) result and analysis:
Molecular marker screening is the result show, has 383 pairs of SSR primers variant between parents.Utilize Datadesk v.5.0 software sick spikelet number and sick joint length that the colony's genotype data of each molecule marker and the scab resistance of its corresponding each family are identified carry out linkage analysis, One-way ANOVA records and the incoherent probability P value of scab resistance and the site contribution rate R to scab resistance 2(table 2), the molecule marker of P＜0.05 are promptly chain with a major gene loci.The genotype of gained molecule marker in colony is by the genotype classification of parent's Wangshuibai and Nanjing University 2419, if the 2419 identical strains of genotype and Nanjing University are that phenotype is anti-, then the localized disease-resistant major gene loci of this mark is from Nanjing University 2419: find the Xgwm107 mark of P=0.0004 and the Xgwm469 mark and anti-expansion major gene loci close linkage of P=0.0089 in Nanjing University 2419, promptly obtain the molecule marker of 2419 anti-expansion major gene loci QFhs.nau-4b of wheat Nanjing University and QFhs.nau-6d1.
Predict the wheat plant resistance by above-mentioned molecular markers for identification major gene loci, expectation can improve the breeding process of China's disease-resistant wheat kind rapidly.
The sequence of table 1. labeled primer and amplified fragments size
Mark The left end primer sequence The right-hand member primer sequence Clip size (bp)
Xgwm107 Xgwm469 ATTAATACCTGAGGGAGGTGC CAACTCAGTGCTCACACAACG GGTCTCAGGAGCAAGAACAC CGATAACCACTCATCCACACC 188 172
The One-way ANOVA of table 2. Nanjing University 2419 anti-expansion major gene locis
Major gene loci Mark Sick spikelet number
P R 2(％)
QFhs.nau-4b Xgwm107 0.0004 17.9
QFhs.nau-6d1 Xgwm469 0.0089 6.1
P: expression and the incoherent probability of scab resistance, P＜0.05 this mark of table and scab resistance close linkage
R 2: the site is to the contribution rate of scab resistance, value is big more show relevant more with scab resistance

Claims (2)

1, the molecule marking method of wheat Nanjing University 2419 anti gibberellic diseases expansion major gene loci is characterized in that:
Use labeled primer Xgwm107,
Left end primer sequence ATTAATACCTGAGGGAGGTGC
Right-hand member primer sequence GGTCTCAGGAGCAAGAACAC
Amplification wheat anti gibberellic disease kind or breeding material DNA, if can amplify the amplified fragments of 188bp, then indicate the existence of the 2419 anti gibberellic diseases expansion major gene loci Qfhs.nau-4b of wheat Nanjing University, wherein this major gene loci is positioned at the 4B karyomit(e) of wheat, utilize Data desk v.5.0 software record with the incoherent probability P value of scab resistance be 0.0004, to the contribution rate 17.9% of scab resistance;
Perhaps use labeled primer Xgwm469,
Left end primer sequence CAACTCAGTGCTCACACAACG
Right-hand member primer sequence CGATAACCACTCATCCACACC
Amplification wheat anti gibberellic disease kind or breeding material DNA, if can amplify the amplified fragments of 172bp, then indicate the existence of the 2419 anti gibberellic diseases expansion major gene loci Qfhs.nau-6d1 of wheat Nanjing University, wherein this major gene loci is positioned at the 6D karyomit(e) of wheat Nanjing University 2419, utilize Data desk v.5.0 software record with the incoherent probability P value of scab resistance be 0.0089, to the contribution rate 6.1% of scab resistance.
2, the molecule marking method of wheat according to claim 1 Nanjing University 2419 anti gibberellic diseases expansion major gene loci is characterized in that the process of screening above-mentioned labeled primer is as follows:
(1) the wheat breed Wangshuibai obtains hybrid F1 as maternal hybridization with male parent Nanjing University 2419, and the F1 selfing produces F2, adopts simple grain transmission method (SSD) to obtain the F6 RIL in generation then;
(2) extract the DNA that each strain of recombinant inbred lines is with the SDS method, adopt simple repeated sequence mark SSR that two parents are carried out the polymorphism screening, PCR carries out on PE9600 amplification instrument, amplified production carries out the electrophoretic separation analysis on the 8%g/ml polyacrylamide gel, according to the molecular marker screening result, filtering out has polymorphic primer between the parent, has polymorphic primer to analyze in recombinant inbred lines between the parent, the pcr amplification program is the same, obtains colony's genotype data;
(3) according to chain exchange rule, utilize colony's genotype data to make up the genetic map of wheat, used software is Mapmaker 2.0, minimum LOD value is made as 3, obtains linkage map;
(4) blooming stage inoculation head blight pathogenic bacteria carries out scab resistance to each family of RIL and identifies, obtains the sick spikelet number and the sick joint length of each family of RIL;
Utilize Data desk v.5.0 software carry out linkage analysis to colony's genotype data of each molecule marker with disease tassel yield that the scab resistance of its corresponding each family is identified, One-way ANOVA records and the incoherent probability P value of scab resistance, the molecule marker of P＜0.05 is promptly chain with a major gene loci, the position of anti gibberellic disease expansion major gene loci is determined by the chromosome position of molecule marker: find the Xgwm107 mark of P=0.0004 and the Xgwm469 mark and anti-expansion major gene loci close linkage of P=0.0089 in Nanjing University 2419, Xgwm107 and Xgwm469 are the labeled primer of 2419 anti-expansion major gene loci QFhs.nau-4b of labeling wheat Nanjing University and QFhs.nau-6d1.
CN 03131961 2003-06-23 2003-06-23 Molecule marker method of wheat Nanda 2419 scab resistant major gene locus Expired - Fee Related CN1246477C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN 03131961 CN1246477C (en) 2003-06-23 2003-06-23 Molecule marker method of wheat Nanda 2419 scab resistant major gene locus

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN 03131961 CN1246477C (en) 2003-06-23 2003-06-23 Molecule marker method of wheat Nanda 2419 scab resistant major gene locus

Publications (2)

Publication Number Publication Date
CN1462807A CN1462807A (en) 2003-12-24
CN1246477C true CN1246477C (en) 2006-03-22

Family

ID=29748489

Family Applications (1)

Application Number Title Priority Date Filing Date
CN 03131961 Expired - Fee Related CN1246477C (en) 2003-06-23 2003-06-23 Molecule marker method of wheat Nanda 2419 scab resistant major gene locus

Country Status (1)

Country Link
CN (1) CN1246477C (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100336905C (en) * 2004-05-08 2007-09-12 南京农业大学 Wheat fertility recovery gene molecular mark and its obtaining method

Also Published As

Publication number Publication date
CN1462807A (en) 2003-12-24

Similar Documents

Publication Publication Date Title
CN108165656B (en) Wheat molecular marker and application thereof in identification of wheat powdery mildew resistance
CN112080582B (en) KASP molecular marker closely linked with major QTL locus of wheat spike length and application thereof
CN105200160B (en) A kind of relevant SNP marker of dissolved oxygen tolerance low to litopenaeus vannamei and its screening and application
Lu et al. Identification of high-efficiency SSR markers for assessing watermelon genetic purity
CN102140506B (en) Molecular marker linked with gummy stem blight resistance gene Gsb-2 and application thereof
CN110578008A (en) molecular marking method for predicting and identifying chicken abdominal fat weight
CN110777216B (en) Method for identifying purity of Jingke waxy 2000 corn hybrid based on SNP marker
CN1246479C (en) Molecule marker method of wheat Wangshuibai scab resistant major gene locus
CN1246478C (en) Molecule marker method of wheat Wangshuibai scab resistant major gene locus
CN112899376B (en) Method for detecting economic traits of Tibetan chicken by FOXO1 gene SNP marker and application thereof
CN105238866B (en) One SNP site related to upland cotton Early mature apricot and its application
CN1246476C (en) Molecule marker method of wheat Nanda 2419 scab resistant major gene locus
Al-Naggar et al. Molecular assessment of genetic diversity among Egyptian landraces of Wheat (Triticum aestivum L.) Using Microsatellite Markers
KR102024212B1 (en) Composition for selecting variety resistant to rice bakanae disease containing qBK1WD gene comprising DNA marker and method for selecting variety resistant to rice bakanae disease using DNA marker
CN1246477C (en) Molecule marker method of wheat Nanda 2419 scab resistant major gene locus
Jayawardana et al. Evaluation of DNA markers linked to blast resistant genes, Pikh, Pit (p), and Pita, for parental selection in Sri Lankan rice breeding
Talebi et al. Molecular characterization of'Carthamus tinctorius' and'C. oxyacanthus' germplasm using sequence related amplified polymorphism (SRAP) markers
de Oliveira et al. Dominant versus codominant marker aiming to characterize Capsicum spp.
CN114231639B (en) Method for identifying or assisting in identifying area of porcine eye muscle and application thereof
CN111485032B (en) Method for identifying cucumber female line and SNP primer combination used by same
CN101942521A (en) Molecular marker detection method of rice blast bacterium non-toxic genes Avr-Pit and primers thereof
CN117757983B (en) Molecular marker primer co-separated from powdery mildew resistance gene PmYD-001 of durum wheat and application thereof
KR101878539B1 (en) Nucleotide polymorphism markers for Foxglove aphid resistance in soybean and the use thereof
Okello et al. Molecular characterization of rice germplasm (Oryza sativa L.) using Simple Sequence Repeat (SSR) markers
CN113201597B (en) InDel molecular marker primer identified by crucifer clubroot-resistant gene PbBa8.1 and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C19 Lapse of patent right due to non-payment of the annual fee
CF01 Termination of patent right due to non-payment of annual fee