CN106676180A - Molecular marker for regulating and controlling maize tassel branch number major QTL (Quantitative Trait Loci) and application thereof - Google Patents
Molecular marker for regulating and controlling maize tassel branch number major QTL (Quantitative Trait Loci) and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of molecular biology, and in particular relates to a molecular marker for regulating and controlling maize tassel branch number major QTL (Quantitative Trait Loci) and application thereof. The molecular marker for regulating and controlling the maize tassel branch number major QTL is formed by two pairs of SSR markers including umc2177 and umc1378. A method for assistantly selecting maize with less tassel branches comprises the steps of extracting genome DNA (Deoxyribonucleic Acid) of the maize to be detected; carrying out PCR (Polymerase Chain Reaction) amplification on the markers umc2177 and umc1378; when obtaining amplified products with the length being 234bp and 331bp, adopting the maize to be detected as candidate maize with less tassel branches, and applying the candidate maize in breeding practice. The maize with less tassel branches is assistantly selected through the molecular marker provided by the invention, so that the number of the tassel branches can be predicted through detecting characteristic stripes of the molecular marker, and an identification method is easy to operate, simple, feasible, and high in selection efficiency, and has great application potentiality in the field of high-yield breeding of maize.
Description
Technical field
The invention belongs to biology field, and in particular to a kind of molecule of regulation and control maize male ears branch amount main effect QTL
Labelling and its application.
Technical background
Used as the important genitals of Semen Maydiss, it not only directly affects the corn seed quality of production, and and Semen Maydiss to tassel
Yield composition is closely related.Numerous studies show the milpa with less tassel, and the light transmission of colony bottom is strong, tassel life
Length is few to the consumption of nutrient, and then improves corn yield.Additionally, scholar's research also finds maize male ears with the tight phase of drought resistance
Close, substantially shortening, tassel branch number are bright for maize male ears index of correlation such as total tassel length and tassel fringe handle length etc. under drought stress
It is aobvious to reduce, take out male spinning time interval (anthesis-silking interval, ASI) and be obviously prolonged, ultimately result in Semen Maydiss tight
The underproduction again.Tassel branch number is the important evaluation index of tassel size, therefore deeply dissects the heredity of maize male ears branching and bounding
Mechanism and its internal relation with drought resistance is disclosed, it is significant to corn with high yield breeding.
Tassel branch number difference is a key character of maize male ears, the tassel branch number of different genetic background storerooms
Differ greatly.Some selfing based material tassel branch numbers are a lot, but the selfing based material tassel branch number having is seldom, or even nothing point
Branch.For Genetic Performance, tassel branch number is that mechanism is extremely complex, and easily receives by the complicated quantitative trait of controlled by multiple genes
Environmental effect, therefore this brings many difficulties to maize male ears study mechanism.In recent years, deeply grinding with molecular biology
The constantly improve with molecular marking technique is studied carefully, using quantitative character gene locus therefor (quantitative trait locus, QTL)
Positioning can substantially determine gene locis and the position of correlated quantitative traitses, estimate the hereditary effect in each site, and then in the future
Molecular breeding is provided fundamental basis and technical support.At present, maize male ears branch amount QTL Position Research has become scholar and breeding
The hot subject of family's concern, but the tassel branch number QTL site with using value is less, therefore, excavate different genetic backgrounds
The tassel branch number main effect QTL of all stable expression has huge potential application valency to corn with high yield breeding under lower and varying environment
Value.
The content of the invention
The present invention is intended to provide a kind of molecular marker of regulation and control maize male ears branch amount main effect QTL, of the invention also to provide auxiliary
The method for selecting the less Semen Maydiss of tassel branch, the present invention is helped also to provide the molecular marker of regulation and control maize male ears branch amount main effect QTL
Application in corn breeding.
In order to solve above-mentioned technical problem, the present invention adopts following three technology scheme：
Firstth, a kind of molecular marker of regulation and control maize male ears branch amount main effect QTL, it is characterised in that：By umc2177 and
Umc1378 two is constituted to primer, and the sequence of the primer umc2177 is：
Forward:5’-ACCATGCATGTCTCACGTCACT-3’
Reverse:5’-GGGTACGTGCTGTGGAGGAC-3’
The sequence of the primer umc1378 is：
Forward:5’-GAAGTCGCTGATGAGAACGTAACC-3’
Reverse:5’-GCTAGCTAGTGTGAGTTCTTCCGC-3’。
The method of the less Semen Maydiss of the secondth, a kind of assisted Selection tassel branch, step is as follows：Extract the genome of Semen Maydiss to be measured
DNA；Enter performing PCR amplification with primer umc2177 and umc1378；When the amplified production that length is 234bp and 331bp is obtained, then treat
It is the less Semen Maydiss of tassel branch to survey Semen Maydiss.
3rd, application of the molecular marker of maize male ears branch amount main effect QTL in corn breeding is regulated and controled, its feature exists
In：The method of the less Semen Maydiss of assisted Selection tassel branch provided with the present invention identifies the less Semen Maydiss of candidate's tassel branch,
Again the less Semen Maydiss of candidate's tassel branch are applied to into corn breeding.
The present invention is by two sets of F2:3The tassel branch number QTL of family is analyzed, and finds under juicy environmental treatment
There is a regulation and control maize male ears branch between umc2177 the and umc1378 labellings in the chromosome Bin7.00 regions of Semen Maydiss the 7th
The main effect QTL is named as qTBN-Ch.7-1 by several main effect QTLs, inventor.Under the process of multiple moisture environments, the qTBN-
Ch.7-1 is in two sets of F2:3The accumulation phenotype contribution rate of family is respectively 46.01% and 34.09%.Analysis shows utilize these two pair
SSR marker enters performing PCR amplification to Semen Maydiss to be measured, and the tassel size of Semen Maydiss to be measured can be predicted.
The beneficial effects of the present invention is：The molecular marker announced by the present invention carries out molecular marker assisted selection, only
Need the feature amplified band of detection molecules labelling, you can prediction maize male ears size, this authentication method is easily operated, simply may be used
Row, efficiency of selection are high.The less com per plant of tassel can be identified in maize growth early stage, eliminate other individual plants, selection target
Clearly, it is and not affected by environment, effectively improve the breeding utilization value of candidate's Semen Maydiss.
Description of the drawings
The histogram of Fig. 1 LTpop tassel branch numbers；
The histogram of Fig. 2 CTpop tassel branch numbers；
Fig. 3 LTpop genetic linkage mapses and tassel branch number main effect QTL positioning schematic diagram；
Fig. 4 CTpop genetic linkage mapses and tassel branch number main effect QTL positioning schematic diagram.
Specific embodiment
With reference to specific embodiment, the present invention will be further described, and test method is such as without special theory in following embodiments
It is bright, conventional methods are, the experiment reagent and consumptive material described in following embodiments if no special instructions, is all from conventional life
Change Reagent Company.
In the present embodiment, the detailed step for obtaining the molecular marker of regulation and control maize male ears branch amount main effect QTL is as follows：
1. Semen Maydiss F2:3The measure of tassel branch number under informative population and different in moisture environmental treatment
3 parts of selfing linies that the tassel branch number and drought resistance obtained with the early stage screening of this seminar differs greatly are as parent's structure
Two sets of F are built2Segregating population, F2Segregating population selfing obtains F2:3Family.Wherein corridor Huang/prosperous 7-2 is that tassel branch number is few and strong
Used as female parent, TS141 is that tassel branch number is more and drought sensitivity selfing line is used as common male parent to drought-enduring selfing line.Corridor Huang and TS141
The F of structure2:3Family is referred to as LTpop, and it includes 202 strains；The F that prosperous 7-2 and TS141 builds2:3Family is referred to as
CTpop, it includes 218 strains.2014 in Wuwei (37.97 ° of N, 102.63 ° of E；1508m) and Zhangye (38.83 ° of N,
106.93°E；1536m) plant LTpop and corresponding parent.2015 in ancient wave (36.67 ° of N, 102.85 ° of E；1785m) and Jingtai
(37.18 ° of N, 104.03 ° of E；1640m) plant CTpop and corresponding parent.Two sets of F2:3Family is all carried out respectively four testing sites
Drought stress process and normal water supply are processed, using flat film drip irrigation technique, by randomized complete-block design, and three repetitions, duplicate rows
Area, the long 6.0m of row, spacing in the rows 0.5m, line-spacing 0.6m.Drought stress process is to terminate to the florescence not filled before the typhon mouth phase
Water, other periods pour water once every 20d.Normal water supply process is poured water in time in hydropenia during corn growth.
Under different in moisture environmental treatment, two sets of F are selected respectively2:3Overall consistent 10 plants of the individual plant of growing way, treats the florescence in family
At the end of determine tassel branch number (tassel branch number, the TBN of each individual plant；TBN is tassel primary branch amount),
Then the tassel size that its meansigma methods represents each strain is taken.According to formula：
Calculate broad-sense heritability (H2), in formula：For genotypic variance,For genotype by environment interaction variance,For
Error, n is environment number, and r is repeat number.2 sets of F2:3Family tassel branch number qualification result is shown in Table 1.Gained LTpop tassels point
The histogram of branch number is as shown in Figure 1；The histogram of gained CTpop tassel branch numbers is as shown in Figure 2.
1 liang of set F of table2:3Family tassel branch number measured value
Table 1The value of tassel branch number(TBN)in two F2:3populations,
respectively
Character explanation in table：W-W, S-W, W-Z, S-Z, W-G, S-G, W-J, S-J are respectively the process of Wuwei normal water supply, force
The process of prestige drought stress, Zhangye normal water supply are processed, Zhangye drought stress is processed, ancient wave normal water supply is processed, the unrestrained arid side of body of Gu
Compel process, the process of Jingtai normal water supply, the process of Jingtai drought stress；It is as follows.
As shown in Table 1：Under same moisture environment is processed, maternal corridor Huang/prosperous 7-2 is poor with the tassel branch number of male parent TS141
It is different notable；Under drought stress process, the tassel branch number of 3 parts of selfing linies is all in reduction trend, but drought sensitivity selfing line TS141
Tassel branch number reduces the significantly larger than strong drought-enduring selfing line corridor Huang/prosperous 7-2 of degree.
From table 1, Fig. 1 and Fig. 2：Under different in moisture environmental treatment, two sets of F of structure2:3The tassel branch number of family is in
Typical normal distribution, and the broad-sense heritability of tassel branch number more greatly 78.69% and 84.26%, show tassel branch number
It is the quantitative trait by controlled by multiple genes, and is affected larger by inheritance, it is feasible to carry out qtl analysis to it.
2.SSR indicia designs and its polymorphism are screened
In Maize genome data base MaizeGDB website (http://www.maizegdb.org/) select to be uniformly distributed
In the SSR marker 872 pairs of 10 chromosomes of Semen Maydiss, Shanghai Sangon synthesis.Using CTAB methods extract yellow parent corridor, prosperous 7-2,
TS141 genomic DNAs, with 1% agarose gel electrophoresiies DNA mass is detected, with the micro spectrophotometer detections of German IMPLEN
DNA concentration.PCR reaction systems are shown in Table 2, and pcr amplification reaction program is shown in Table 3, and amplified production is solidifying with 8% non-denaturing polyacrylamide
Gel electrophoresis, silver staining.
Jing is analyzed, and 213 pairs of SSR markers that band is clear, polymorphism is good is filtered out between parent corridor Huang and TS141, in parent
217 pairs of SSR markers that band is clear, polymorphism is good are filtered out between this prosperous 7-2 and TS141.These polymorphism SSR markers will be used for
Two sets of F2Segregating population gene type assay, and build corresponding genetic linkage mapses into step 3.
Table 2PCR reaction systems
Table 1The PCR reaction system
Table 3PCR amplified reaction programs
Table 2The PCR amplification reaction process
3. genetic linkage mapses build
The two sets of F built using step 12The corresponding polymorphism SSR marker that segregating population and step 2 are filtered out, to two
Cover corresponding F2Segregating population carries out gene type assay, and using JoinMap4.0 software (http://www.kyazma.nl/
Index.php/mc.JoinMap/sc.Evaluate/) build genetic linkage mapses, using Kosambi functions calculate heredity away from
From (centimorgan, cM).The genetic linkage mapses of gained LTpop are shown in Fig. 3；The genetic linkage mapses of gained CTpop are shown in Fig. 4.
From Fig. 2 and Fig. 3：The two sets of genetic linkage mapses for building respectively include respectively 199 and 205 pairs of SSR markers, cover
10 linkage groups of Semen Maydiss, total genetic distance is 1542.5cM and 1648.8cM, and average genetic is between molecular marker
7.8cM and 8.0cM.Two sets of genetic maps and IBM2 2008Neighbors (http://www.maizegdb.org/data_
Center/map) compare, be marked at highly consistent with reference to the relative ranks on collection of illustrative plates.
4. tassel branch number main effect QTL is positioned
According to two sets of F2:3Tassel branch number phenotypic number of the family under different in moisture environmental treatment, using Windows QTL
Software (the http of Cartographer version 2.5://statgen.ncsu.edu/qtlcart/WQTLcart.htm) in
Composite interval mapping method (composite interval mapping, CIM) detect corresponding F2:3The tassel branch number of family
QTL.For CIM, using Zmapqtl program modules Model 6, window size is 10.0cM, every 0.5cM to tassel branch
Number carries out genotype scanning, and by 1000 sampling LOD threshold value (LOD are determined>3.0).According to dominant effect (D) and additive effect
(A) absolute value of ratio is estimating QTL model of action (Stuberet al.1987)：| D/A |=0.00~0.20 is additivity
(A), | D/A |=0.21~0.80 be partial dominance (PD), | D/A |=0.81~1.20 be dominant (D), | D/A |>1.20 it is super
Dominant (OD).The naming method of QTL names reference McCouch et al. (1997), i.e. q+ character title abbreviation+chromosome name
Claim+QTL sequence numbers on chromosome.Using the Software Create main effect tassel branch number QTL two ends SSR of Patentin version 3.5
Labelled sequence table；Main effect tassel branch number QTL two ends SSR marker sequence table is at description end.Gained LTpop tassel branch numbers
Main effect QTL positioning schematic diagram is shown in Fig. 3；Gained CTpop tassel branch numbers main effect QTL positioning schematic diagram is shown in Fig. 4.Two sets of F2:3Family
Tassel branch number main effect QTL testing result is shown in Table 4.
4 liang of set F of table2:3Family tassel branch number QTL is detected
Table 4The QTL analysis for tassel branch number(TBN)in two F2: 3populations,respectively
Jing CIM analysis shows, from table 4, Fig. 3 and Fig. 4, two sets of F2:3Family is detected simultaneously under multiple moisture environments
To be named as qTBN-Ch.7-1, it is located at the 7th chromosome Bin 7.00 to main effect QTL to a regulation and control tassel branch number
Place, between labelling umc2177 and umc1378, the model of action of gene reduces tassel branch number based on expressed superdominance effect
Allele derive from parent corridor Huang/prosperous 7-2.The QTL is in the case where 2 drought stresses are processed and 1 normally supplies in LTpop
It is detected simultaneously under water process, its accumulation phenotype contribution rate is 46.01%, and the genetic distance is 19.7cM.Should in CTpop
QTL is detected simultaneously in the case where 2 drought stresses are processed and under 1 normal water supply is processed, and its accumulation phenotype contribution rate is
34.19%, the genetic distance is 0.6cM.Show the main effect QTL that qTBN-Ch.7-1 is regulation and control maize male ears branch amount, and and drought resisting
Property it is closely related, the QTL reduces the allele of tassel branch number and derives from parent corridor Huang/prosperous 7-2, therefore can be used for Semen Maydiss
Tassel size is predicted, but because the gene action mode of the QTL is superdominance, therefore especially to note when cenospecies are assembled
Impact of the meaning hybrid vigor to tassel size.
The molecular marker of above-mentioned regulation and control maize male ears branch amount main effect QTL, by umc2177 and umc1378 two to SSR marker
Constitute, the sequence of wherein labelling umc2177 is：
Forward:5’-ACCATGCATGTCTCACGTCACT-3’
Reverse:5’-GGGTACGTGCTGTGGAGGAC-3’
The sequence of the primer umc1378 is：
Forward:5’-GAAGTCGCTGATGAGAACGTAACC-3’
Reverse:5’-GCTAGCTAGTGTGAGTTCTTCCGC-3’
Using the molecular marker assisted selection tassel branch less Semen Maydiss of above-mentioned regulation and control maize male ears branch amount main effect QTL
Method includes：Corn gene group DNA to be measured is extracted, with labelling umc2177 and umc1378 performing PCR amplification is entered, be when length is obtained
The amplified production of 234bp and 331bp, then Semen Maydiss to be measured are the less com per plant of candidate's tassel branch number, eliminate other individual plants,
It is autotelic to assemble cross combination, select tassel be of moderate size, high-yield corn new varieties, and then be applied in production practices.
Sequence table
SEQUENCE LISTING
<110>Gansu Agriculture University
<120>The molecular marker of regulation and control maize male ears branch amount main effect QTL and its application
<130> 2016
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> DNA
<213>Zea Semen Maydiss（Zea mays L.)
<400> 1
accatgcatg tctcacgtca ct 22
<210> 2
<211> 20
<212> DNA
<213>Zea Semen Maydiss（Zea mays L.）
<400> 2
gggtacgtgc tgtggaggac 20
<210> 3
<211> 24
<212> DNA
<213>Zea Semen Maydiss（Zea mays L.）
<400> 3
gaagtcgctg atgagaacgt aacc 24
<210> 4
<211> 24
<212> DNA
<213>Zea Semen Maydiss（Zea mays L.）
<400> 4
gctagctagt gtgagttctt ccgc 24
1
Claims (3)
1. it is a kind of regulation and control maize male ears branch amount main effect QTL molecular marker, it is characterised in that：By molecular marker umc2177 and
Umc1378 is constituted, and the primer sequence of the molecular marker umc2177 is：
Forward:5’-ACCATGCATGTCTCACGTCACT-3’；
Reverse:5’-GGGTACGTGCTGTGGAGGAC-3’；
The sequence of the primer umc1378 is：
Forward:5’-GAAGTCGCTGATGAGAACGTAACC-3’；
Reverse:5’-GCTAGCTAGTGTGAGTTCTTCCGC-3’。
2. a kind of method of the less Semen Maydiss of assisted Selection tassel branch, step is as follows：Extract the genomic DNA of Semen Maydiss to be measured；With
Molecular labeling primer sequence umc2177 and umc1378 described in claim 1 enters performing PCR amplification；It is 234bp when length is obtained
With the amplified production of 331bp, then Semen Maydiss to be measured are the less Semen Maydiss of tassel branch.
3. described in a kind of claim 1 regulation and control maize male ears branch amount main effect QTL molecular marker in corn breeding should
With, it is characterised in that：The method of the less Semen Maydiss of assisted Selection tassel branch provided with claim 2 identifies candidate's tassel point
The less Semen Maydiss of branch, then the less Semen Maydiss of candidate's tassel branch are applied to into corn breeding.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110923353A (en) * | 2019-12-02 | 2020-03-27 | 甘肃农业大学 | Molecular marker for regulating and controlling main effect QTL (quantitative trait locus) of photosynthetic property of corn and application of molecular marker |
| CN112029897A (en) * | 2020-10-14 | 2020-12-04 | 江苏省农业科学院 | SNP marker closely linked with continuous multi-leaf-position leaf width main effect QTL under corn tassel and application thereof |
| CN112626261A (en) * | 2021-01-18 | 2021-04-09 | 吉林大学 | SNP molecular marker related to maize tassel branch number character and application |
-
2017
- 2017-01-22 CN CN201710045421.XA patent/CN106676180B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| MARY POLCCO ET AL.: "Missouri maize project releases new SSR marker data", 《HTTP://WWW.BIO.NET/BIONET/MM/MAIZE/2000-MARCH/000746.HTML》 * |
| N.UPADYAYULA ET AL.: "Genetic and QTL analysis of maize tassel and ear inflorescence architecture", 《THEOR APPL GENET》 * |
| QIANQIAN ZHANG ET AL.: "Association analysis of important agronomical traits of maize inbred lines with SSRs", 《AUSTRALIAN JOURNAL OF CROP SCIENCE》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110923353A (en) * | 2019-12-02 | 2020-03-27 | 甘肃农业大学 | Molecular marker for regulating and controlling main effect QTL (quantitative trait locus) of photosynthetic property of corn and application of molecular marker |
| CN112029897A (en) * | 2020-10-14 | 2020-12-04 | 江苏省农业科学院 | SNP marker closely linked with continuous multi-leaf-position leaf width main effect QTL under corn tassel and application thereof |
| CN112626261A (en) * | 2021-01-18 | 2021-04-09 | 吉林大学 | SNP molecular marker related to maize tassel branch number character and application |
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| CN106676180B (en) | 2020-03-31 |
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