CN106399468B - Rice early heading main effect QTL molecular labeling and its identification method and application - Google Patents
Rice early heading main effect QTL molecular labeling and its identification method and application Download PDFInfo
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Abstract
The present invention discloses a kind of rice early heading main effect QTL molecular labeling and its identification method and application, have found rice early heading main effect QTL qHD8.3 molecular labeling, it is RM6670, M55, M07, M13 and M15 respectively, using one or more pairs of carry out PCR amplifications in the corresponding primer of above-mentioned molecular labeling, amplified production is detected;The present invention also provides the application of above-mentioned molecular labeling or molecular marker identification method in the rice early positioning and clone of heading QTL qHD8.3.It can accelerate the clone of morning heading QTL qHD8.3 through the invention, and assisted selection can be applied to, convenient for timely hybridizing transformation, accelerate breeding process.
Description
Technical field
The present invention relates to molecular biology field, early heading main effect QTL molecular labeling and its identification of especially a kind of rice
Methods and applications.
Background technique
Rice is as one of most important cereal crops in China or even the world, and the raising of yield is for solving the following whole world
Food problem has highly important strategic importance.However, mostly there is extremely strong region and seasons for existing rice varieties
Limitation, this largely affects further increasing for the Main Agronomic Characters such as yield and quality, therefore improves rice
The region of kind and seasonal adaptation cultivate the new rice variety for adapting to given area ecological environment and cultivating cropping system, at
For one of the main target of current rice breeding.
Heading stage is the Main Agronomic Characters for determining rice varieties region and seasonal adaptation, to the high yields of rice varieties and
Stable yields plays an important role, for a long time by the extensive attention of breeder.Rice ear sprouting period mainly by its photonasty, response to temperature and
Basic nutrition growth determines (Chang TT, Li CC, Vergara BS. (1969) Euphytica, 18:79-91;Tsai
KH.(1985)Rice Genet Newslett,2:77-78;Tsai KH.(1986)In:Rice
Genetics.International Rice Research Institute,339-349).Due to the light temperature item of different regions
There are great differences for part, thus the improvement of different regions precocity rice varieties have to select different types of early heading QTL with
Adapt to local light and temperature condition.Although being cloned into much genes with arabidopsis florescence DNA homolog in current rice,
But there is very big differences for their Regulation Mechanisms to the florescence;Moreover, although the method with homologous clone obtains in rice
It so that rice is obtained the characteristic early eared after the early heading gene overexpression obtained, but due to the difference of genetic background, cause
These homologous genes can not still be applied to the cultivating process of precocious new rice variety so far.Therefore, rice early heading gene is excavated,
It is early in the Poa linn kind for adapting to South China Double Cropping rice workspace ecological condition and its cross combination, solution breeding practice to cultivating
The ripe contradiction for being difficult to take into account with high yield is of great significance.
In terms of the positional cloning research of Heading date gene, up to the present, there are a large amount of heading stage QTLs positioning
Report, but less by the heading stage QTL that the method for map based cloning is excavated, so far, only Hd1, Hd6, Hd3a,
Ehd1, Ehd2, Ghd7, DTH8 and EL1 (Yano M.et al. (2000) Plant Cell, 12:2473-2483;
Takahashi,Y.et al.(2001)Proc Natl Acad Sci USA,98:7922-7927;Kojima S.et al.
(2002)Plant Cell Physiol,43:1096–1105;Doi K.et al.(2004)Genes Dev,18:926-936;
Matsubara K.et al.(2008)Plant Physiol,148(3):1425-1435;Xue WY.et al.(2007)Nat
Genet,40:761-767;Wei XJ.et al.(2010)Plant Physiol,153:1747-1758;Dai C,Xue HW.
(2010) Embo J, 29 (11): 1916-1927) etc. genes be to be cloned by the method for map based cloning, the clone of these genes
It is greatly promoted understanding of the people to Rice Heading mechanism.
Due to the excavation of heading stage QTL, our understanding and understanding to Rice Heading regulated and control network can be deepened, meanwhile,
Its molecular labeling obtained can be also used for assisted selection.Due to molecular marker assisted selection (Marker-Assisted
Selection, MAS) method is the selection carried out to the genotype of kind, be not protected from environmental, thus MAS Breeding Application in
Improvement to rice morning heading QTL, will effectively promote the efficiency of breeding.
Summary of the invention
The present invention provides a kind of early heading main effect QTL molecular labeling and its identification method and the applications of rice, pass through detection
With rice morning heading main effect QTL molecular labeling, it can determine that whether there is or not early heading channel genes into breeding lines, improve the character
Efficiency of selection, accelerate Breeding progress.
To achieve the above object, the technical solution of the present invention is as follows:
A kind of early heading main effect QTL molecular labeling of rice, is named as qHD8.3, the molecular labeling be respectively RM6670,
M55, M07, M13 and M15, corresponding primer is respectively:
RM6670:
Upstream primer: 5'-TCCGTCGCTGCAAATTAGTCC -3';
Downstream primer: 5'-CCATGTACCTCTATAATGGCAAGACC -3';
M55:
Upstream primer: 5'-CCATTTGGTAGGTCCATCTTACCC -3';
Downstream primer: 5'-CTCCCAAGTGAAGTGCTGTCTGG -3';
M07:
Upstream primer: 5'-GGTGGTCTTGATTCCCTTGT -3';
Downstream primer: 5'-GAAACAGATCAGCCTCACTG -3';
M13:
Upstream primer: 5'-ATATACAAGCGAACTCCTGT -3';
Downstream primer: 5'-TAAGTGCGATGAAACAATGA -3';
M15:
Upstream primer: 5'-TGGCACCATCCTCATTGCTC -3';
Downstream primer: 5'-AGCGTCGTCAGATACATTGG -3';
The acquisition methods of above-described rice morning heading main effect QTL molecular labeling, comprising the following steps:
Using DP30 as donor parents, with rice variety 9311 for receptor parent and recurrent parent, pass through continuous backcross, selfing
And M8003 line, in BC4F2In generation, obtains the early heading chromosome segment substitution line CSSL13 for stablizing heredity;Again by substitution line
CSSL13 hybridizes with 9311, obtains F1Cenospecies obtains F after selfing2Segregating population selects F2Heading stage shows as morning in group
Multiple single plants of phase or advanced stage, plant into F respectively2:3Strain selects F2:3The correspondence F that strain does not separate2Single plant is for chain
Analysis, obtains molecular labeling described in claim 1.
The present invention also provides rice early to ear main effect QTL molecular labeling in the rice early positioning and clone of heading QTL
Application.
Above-described rice morning heading main effect QTL molecular marker identification method, comprising the following steps:
Using rice material complete genome DNA to be identified as template, using above-described molecular labeling RM6670, M55,
One or more pairs of carry out PCR amplifications in M07, M13 and M15 corresponding primer, detect amplified production:
(1) it is expanded using molecular labeling RM6670 primer, if amplifying the amplified fragments of 177bp, shows there is morning
The presence of heading main effect QTL;
(2) it is expanded using molecular labeling M55 primer, if amplifying the amplified fragments of 177bp, shows there is early heading
The presence of main effect QTL;
(3) it is expanded using molecular labeling M07 primer, if amplifying the amplified fragments of 166bp, shows there is early heading
The presence of main effect QTL;
(4) it is expanded using molecular labeling M13 primer, if amplifying the amplified fragments of 43bp, shows there is early heading
The presence of main effect QTL;
(5) it is expanded using molecular labeling M15 primer, if amplifying the amplified fragments of 131bp, shows there is early heading
The presence of main effect QTL.
Wherein, the reaction system of PCR described above is as follows:
The reaction condition of PCR described above are as follows: 94 DEG C of denaturation 5min, 94 DEG C of 30s, 56 DEG C of 30s, 72 DEG C of 60s expand 35
Circulation, last 72 DEG C extend 10min eventually.
Above-described molecular marker identification method, DNA cloning product is detected through polyacrylamide gel electrophoresis, and is passed through
The DNA band of silver staining colour developing record amplification.
The object of the invention is also to provide above-described molecular marker identification methods in breeding rice morning heading product
Application in kind.
The invention has the benefit that
(1) present invention finds a kind of rice morning heading main effect QTL qHD8.3 molecular labelings, with the rice being previously reported
Early heading QTL such as Hd1, Hd6, Hd3a, Ehd1, Ehd2, Ghd7 and DTH8 etc. are different, enrich people and regulate and control to Rice Heading
The understanding and understanding of network also increase the diversity of rice morning heading genetic resources, to cultivate different types of early heading water
Rice varieties provide more choices;
(3) present invention can accelerate morning heading QTL by the discovery of rice morning heading main effect QTL qHD8.3 molecular labeling
The clone of qHD8.3, and assisted selection can be applied to, convenient for timely hybridizing transformation, accelerate breeding process;
(2) present invention establishes a kind of rice early heading main effect QTL qHD8.3 molecular marker identification method, can it is accurate,
Rapidly identify the early heading single plant with qHD8.3, improves the efficiency of selection of the character, accelerate Breeding progress.
Detailed description of the invention
Fig. 1 is the heading stage phenotype of chromosome segment substitution line CSSL13;Wherein: Fig. 1 a: under normal long-day conditions
Difference in 9311 and CSSL13 phenotype;Fig. 1 b:2014 early season counts 9311 and CSSL13 each 30 plants of heading stage, calculates flat
Mean value, A, B indicate 0.01 horizontal lower significant difference;Fig. 1 c:2014 season in evening counts 9311 and CSSL13 each 30 plants of heading stage,
Average value is calculated, C, D indicate 0.01 horizontal lower significant difference.
Fig. 2 is F2For single plant heading stage distribution map.
Fig. 3 is molecular labeling primer (M07 and M13) to F2The partial results figure of 52 evening heading single plant amplifications of generation;Wherein,
Fig. 3 a: molecular labeling primer M07 to evening heading single plant amplification banding pattern;Fig. 3 b: molecular labeling primer M13 to evening heading single plant
Banding pattern is expanded, the 10th plant is exchange single plant;
Fig. 4 is Primary Location figure of the rice qHD8.3 gene on the 8th chromosome.
Specific embodiment
Below with reference to embodiment, the invention will be further described, but embodiments of the present invention be not limited to
Lower embodiment.
Embodiment 1: the screening and positioning of molecular labeling
(1) building of early heading chromosome segment substitution line CSSL13 and replacement fragment analysis
Using 9311 be receptor parent, Guangxi common wild-rice DP30 is donor parents, by continuous backcross, selfing and orient
Selection, in BC4F2What generation obtained stablizes the early heading chromosome segment substitution line CSSL13 of heredity.
The SSR marker and Indica announced according to the website gramene (http://www.gramene.org/) obstruct sequence
The STS of difference designed, designed is marked, and to polymorphic detection is carried out between parent DP30 and 9311, detects that 408 pairs show altogether
The label of obvious polymorphism, carries out replacement fragment analysis to CSSL13 using this 408 pairs of polymorphic molecular markers, the results showed that,
Replacement segment of the CSSL13 altogether containing 5 donor parents DP30 is located on the 1st, 2,3,8 and No. 11 chromosome, remaining back
Scape is all from 9311 (tables 1), and the presence of these replacement segments may be the reason of causing its heading stage to make a variation.
The replacement segment and its length of table 1CSSL13
Number | Chromosome | Replace segment | Length (kb) |
SL1 | 1 | RM486-RM11865-RM315 | 1778 |
SL2 | 2 | RM7423-M9-RM13429-RM1920 | 6765 |
SL3 | 3 | M11-RM15288-RM411 | 3850 |
SL4 | 8 | RM6670-M55-M07-M13-M15 | 5713 |
SL5 | 11 | M230-RM6680 | 1792 |
(2) the Genetic characteristics specificity analysis of substitution line CSSL13
Under the normal short-day in Nanning (Nanning early season) and the field condition of normal long-day (Nanning season in evening), CSSL13 points
Do not ear ahead of time 14.2d (Fig. 1 a, Fig. 1 b) and 13.8d (Fig. 1 c) than receptor parent 9311, t test Analysis show CSSL13 and
Heading stage difference between 9311 has all reached extremely significant level.This shows that the characteristic that CSSL13 is early eared may be to illumination and temperature
It spends insensitive.(table is handled by illumination box stringent long-day and short-day, high temperature and low-temperature treatment with E1 and E2 respectively
Show), as shown in table 2, the results showed that the heading stage difference between CSSL13 and 9311 be respectively 14.5d, 14.8d, 14.0d and
13.0d, difference have all reached extremely significant horizontal (table 2), have further demonstrated that qHD8.3 is an early pumping insensitive to light temperature
Fringe QTL.
The heading stage number of days of table 2 substitution line material C SSL13 and 9311 under difficult environmental conditions
Note: E1: high temperature long-day environment;E2: shading treatment, 10h illumination/14h are dark;E3: 32 DEG C of daytime, night 27
DEG C, 12h illumination/12h is dark;E4: on 25 DEG C of daytime, 20 DEG C of night, 12h illumination/12h is dark.A: 0.05 horizontal lower difference is indicated
It is not significant
(3) F of CSSL13/93112The building and genetic analysis of segregating population
(1) hybridized using early heading chromosome segment substitution line CSSL13 with receptor parent 9311, obtain F1Cenospecies, kind
Plant F1Cenospecies, selfing obtain F2Segregating population is used for genetic analysis.
(2) to F2Segregating population carries out early heading identification
Result is recorded according to field investigation to draw, the results showed that, in F2In 412 single plants of segregating population, heading stage is
76~100d, as shown in Fig. 2, apparent bimodal distribution is presented.Using 92d as boundary, group is divided into early heading and evening heading two is big
Class.The single plant early eared has 297 plants, and the evening single plant of heading has 115 plants.Through analysis shows, the single plant number of early heading and evening heading
Mesh meets the segregation ratio (X of 3:12 c=0.94 < X2 0.05,1=3.841), this shows that CSSL13 early ears characteristic by a pair of dominant
The control of main effect karyogene.
(3) qHD8.3 molecular labeling linkage analysis and Primary Location
From F2In 412 single plants of segregating population, 100 plants of early heading single plants and 100 plants of evening heading single plants are chosen, it is single respectively
Strain sowing and plantation, to the F of these single plants2:3Strain carries out descendant's detection, and finishing screen selects 76 morning for stablizing heredity at heading stage
Heading single plant and 52 evenings heading single plants are used for the Primary Location of target gene.
Using the polymorphic molecular marker with genetic background screened are stablized at these heading stages with the single plant of heredity
Linkage analysis is carried out, as a result, it has been found that, there are chain, two labels by the molecular labeling RM6670 and M15 on qHD8.3 and No. 8 chromosome
Locate 11 and 5 plants of exchange single plants of each appearance, and mark 11 plants on the 5 plants of exchange single plants and label RM6670 on M15 to be different from,
Show that target gene is likely located between label RM6670 and M15;For the position for further reducing simultaneously hard objectives gene, continue
Using label RM6670 and M15 between three couples of molecular labelings M55, M07 and M13, to stablize heredity early, late heading single plant into
Row PCR amplification and electrophoresis detection, the results showed that, also there are 6 plants, 2 plants, 1 plant of exchange list respectively in molecular labeling M55, M07 and M13
Strain, and 6 plants of exchange single plants on molecular labeling M55 are included in 11 plants of exchange single plants of molecular labeling RM6670, molecule mark
2 plants of exchange single plants on M07 are remembered in 6 plants of exchange single plants of molecular labeling M55, and 5 plants of exchanges on molecular labeling M15 are single
Strain contains 1 plant of exchange single plant on molecular labeling M13, and not with the exchange single plant on molecular labeling RM6670, M55, M07
It repeating, it was confirmed that the gene is located on the 8th chromosome, and is positioned between molecular labeling M07 and M13, as shown in Figure 3a, point
For sub- labeled primer M07 to the amplification banding pattern of evening heading single plant, the 20th plant is exchange single plant, as shown in Figure 3b, molecular labeling primer
For M13 to the amplification banding pattern of evening heading single plant, the 10th plant is exchange single plant;As shown in connection with fig. 4, physical distance is about 1.6Mb.
Above-mentioned linkage analysis and Primary Location process, used DNA extraction method and PCR reaction condition are respectively as follows:
A. donor parents, receptor parent and F are extracted with conventional CTAB method2The DNA of each single plant of segregating population;
The reaction system of B.PCR is as follows:
PCR reaction condition are as follows: 94 DEG C of denaturation 5min, 94 DEG C of 30s, 56 DEG C of 30s, 72 DEG C of 60s expand 35 circulations, finally
72 DEG C extend 10min eventually.
DNA cloning product is detected through polyacrylamide gel electrophoresis, is developed the color by silver staining and is recorded the DNA band of amplification.
By the above process it can be seen that can effectively be opened using the molecular labeling of these and early heading main effect QTL qHD8.3
The early heading QTL molecular marker assisted selection breeding of exhibition;It can use big segregating population simultaneously, accelerate the essence to early heading QTL
Fine positioning and map based cloning.
(4) F2:3Strain offspring screens analysis
Using the molecular labeling M07 and M13 of acquisition, in F2:376 heterozygosis single plant building F have been screened in strain offspring3It is fixed
Position group, heterozygosis single plant divide single plant harvest plantation, and the separation of heading stage phenotype occurs in each strain, and early heading and evening take out
The segregation ratio of fringe single plant meets the ratio of 3:1, and screening accuracy rate reaches 100%.Illustrate the molecule using qHD8.3 new gene
It is effective for marking M07 and M13 to carry out the marker assisted selection of rice ear sprouting period.
(5) interpretation of result
The above results show using RM6670, M55, M07, M13 and M15 can early heading main effect QTL carries out molecule to rice
Label, corresponding primer such as table 3:
The primer and its sequence that table 3 positions
2 molecular marker identification method of embodiment
Using rice material complete genome DNA to be identified as template, using above-described molecular labeling RM6670, M55,
One or more pairs of carry out PCR amplifications in M07, M13 and M15 corresponding primer, detect amplified production.
(1) donor parents, receptor parent and F are extracted with conventional CTAB method2The DNA of each single plant of segregating population;
(2) reaction system of PCR is as follows:
PCR reaction condition are as follows: 94 DEG C of denaturation 5min, 94 DEG C of 30s, 56 DEG C of 30s, 72 DEG C of 60s expand 35 circulations, finally
72 DEG C extend 10min eventually.
DNA cloning product is detected through polyacrylamide gel electrophoresis, is developed the color by silver staining and is recorded the DNA band of amplification.
(3) interpretation of result
(1) it is expanded using molecular labeling RM6670 primer, if amplifying the amplified fragments of 177bp, shows there is morning
The presence of heading main effect QTL shows the amplified fragments for having parent 9311 if the amplified fragments of 168bp can be amplified;
(2) it is expanded using molecular labeling M55 primer, if amplifying the amplified fragments of 177bp, shows there is early heading
The presence of main effect QTL shows the amplified fragments for having parent 9311 if the amplified fragments of 169bp can be amplified;
(3) it is expanded using molecular labeling M07 primer, if amplifying the amplified fragments of 166bp, shows there is early heading
The presence of main effect QTL shows the amplified fragments for having parent 9311 if the amplified fragments of 150bp can be amplified;
(4) it is expanded using molecular labeling M13 primer, if amplifying the amplified fragments of 43bp, shows there is early heading
The presence of main effect QTL shows the amplified fragments for having parent 9311 if the amplified fragments of 52bp can be amplified;
(5) it is expanded using molecular labeling M15 primer, if amplifying the amplified fragments of 131bp, shows there is early heading
The presence of main effect QTL shows the amplified fragments for having parent 9311 if the amplified fragments of 117bp can be amplified.
Rice varieties are detected based on above-mentioned 5 molecular labelings, it may be verified that whether contain the presence of early heading main effect QTL,
The breeding that can carry out early rice varieties of earing, to accelerate Breeding progress.
Sequence table
<110>Guangxi University
<120>rice early heading main effect QTL molecular labeling and its identification method and application
<160>10
<210>1
<211>21
<212>DNA
<213>artificial sequence
<221>molecular labeling RM6670 upstream primer
<400>1
TCCGTCGCTGCAAATTAGTCC 21
<210>2
<211>26
<212>DNA
<213>artificial sequence
<221>molecular labeling RM6670 downstream primer
<400>2
CCATGTACCTCTATAATGGCAAGACC 26
<210>3
<211>26
<212>DNA
<213>artificial sequence
<221>molecular labeling M55 upstream primer
<400>3
CCATTTGGTAGGTCCATCTTACCC 24
<210>4
<211>26
<212>DNA
<213>artificial sequence
<221>molecular labeling M55 downstream primer
<400>4
CTCCCAAGTGAAGTGCTGTCTGG 23
<210>5
<211>20
<212>DNA
<213>artificial sequence
<221>molecular labeling M07 upstream primer
<400>5
GGTGGTCTTGATTCCCTTGT 20
<210>6
<211>20
<212>DNA
<213>artificial sequence
<221>molecular labeling M07 downstream primer
<400>6
GAAACAGATCAGCCTCACTG 20
<210>7
<211>20
<212>DNA
<213>artificial sequence
<221>molecular labeling M13 upstream primer
<400>7
ATATACAAGCGAACTCCTGT 20
<210>8
<211>20
<212>DNA
<213>artificial sequence
<221>molecular labeling M13 downstream primer
<400>8
TAAGTGCGATGAAACAATGA 20
<210>9
<211>20
<212>DNA
<213>artificial sequence
<221>molecular labeling M15 upstream primer
<400>9
TGGCACCATCCTCATTGCTC 20
<210>10
<211>20
<212>DNA
<213>artificial sequence
<221>molecular labeling M15 downstream primer
<400>10
AGCGTCGTCAGATACATTGG 20
Claims (7)
- The acquisition methods of main effect QTL molecular labeling 1. a kind of rice is early eared, it is characterised in that the following steps are included:Using DP30 as donor parents, with rice variety 9311 for receptor parent and recurrent parent, by continuous backcross, it is selfed and determines To selection, in BC4F2In generation, obtains the early heading chromosome segment substitution line CSSL13 for stablizing heredity;Again by substitution line CSSL13 with 9311 hybridization, obtain F1Cenospecies obtains F after selfing2Segregating population selects F2Heading stage shows as early stage or advanced stage in group Multiple single plants, plant into F respectively2:3Strain selects F2:3The correspondence F that strain does not separate2Single plant is used for linkage analysis, obtains Following rice morning heading main effect QTL molecular labeling:It is RM6670, M55, M07, M13 and M15 respectively, corresponding primer is respectively:RM6670:Upstream primer: 5'-TCCGTCGCTGCAAATTAGTCC -3';Downstream primer: 5'-CCATGTACCTCTATAATGGCAAGACC -3';M55:Upstream primer: 5'-CCATTTGGTAGGTCCATCTTACCC -3';Downstream primer: 5'-CTCCCAAGTGAAGTGCTGTCTGG -3';M07:Upstream primer: 5'-GGTGGTCTTGATTCCCTTGT -3';Downstream primer: 5'-GAAACAGATCAGCCTCACTG -3';M13:Upstream primer: 5'-ATATACAAGCGAACTCCTGT -3';Downstream primer: 5'-TAAGTGCGATGAAACAATGA -3';M15:Upstream primer: 5'-TGGCACCATCCTCATTGCTC -3';Downstream primer: 5'-AGCGTCGTCAGATACATTGG -3'.
- 2. rice as described in claim 1 early positioning and clone of the heading main effect QTL molecular labeling in rice morning heading QTL In application.
- The identification method of main effect QTL molecular labeling 3. a kind of rice is early eared, it is characterised in that the following steps are included:Using rice material complete genome DNA to be identified as template, using described in claim 1 molecular labeling RM6670, One or more pairs of carry out PCR amplifications in M55, M07, M13 and M15 corresponding primer, detect amplified production:(1) it is expanded using molecular labeling RM6670 primer, if amplifying the amplified fragments of 177bp, shows there is early heading The presence of main effect QTL;(2) it is expanded using molecular labeling M55 primer, if amplifying the amplified fragments of 177bp, shows there is early heading main effect The presence of QTL;(3) it is expanded using molecular labeling M07 primer, if amplifying the amplified fragments of 166bp, shows there is early heading main effect The presence of QTL;(4) it is expanded using molecular labeling M13 primer, if amplifying the amplified fragments of 43bp, shows there is early heading main effect The presence of QTL;(5) it is expanded using molecular labeling M15 primer, if amplifying the amplified fragments of 131bp, shows there is early heading main effect The presence of QTL.
- The identification method of main effect QTL molecular labeling 4. rice as claimed in claim 3 is early eared, it is characterised in that:The reaction system of the PCR is as follows:2.5 μ L of primer 4pmol/ μ LdNTP 2.5mmol/L 2.0μL1.0 μ L of DNA profiling 25ng/ μ L0.2 μ L of Taq archaeal dna polymerase 5U/ μ L10 × PCR buffer, 1.0 μ LddH2O 3.3μL。
- The identification method of main effect QTL molecular labeling 5. rice as claimed in claim 3 is early eared, it is characterised in that:PCR reaction condition: 94 DEG C of denaturation 5min, 94 DEG C of 30s, 56 DEG C of 30s, 72 DEG C of 60s expand 35 circulations, and last 72 DEG C Extend 10min eventually.
- The identification method of main effect QTL molecular labeling 6. rice as claimed in claim 3 is early eared, it is characterised in that:DNA cloning product is detected through polyacrylamide gel electrophoresis, and is developed the color by silver staining and recorded the DNA band of amplification.
- 7. as early heading main effect QTL molecular marker identification method is early in breeding rice for the described in any item rice of claim 3 ~ 6 Application in kind of earing.
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CN106834524B (en) * | 2017-03-31 | 2020-03-17 | 湖南杂交水稻研究中心 | Positioning of rice panicle length major QTLPL6-5 and molecular marker linked with major QTLPL6-5 |
CN108456744B (en) * | 2018-05-15 | 2021-08-06 | 中垦种业股份有限公司 | Molecular marker of valuable rice storage-tolerant QTL and breeding application |
CN111206113B (en) * | 2020-02-12 | 2021-07-02 | 广西壮族自治区农业科学院 | InDel molecular marker for assisting selection of early heading genes of rice and application of InDel molecular marker |
CN111635958B (en) * | 2020-07-22 | 2022-05-24 | 中国农业科学院作物科学研究所 | Molecular marker linked with rice cold-resistant gene qSF12 and application thereof |
CN112501341B (en) * | 2020-12-09 | 2022-05-03 | 浙江师范大学 | Major QTL for regulating heading stage of rice, molecular marker and application |
CN113215296B (en) * | 2021-04-28 | 2022-08-09 | 广西大学 | Rice awn length genegna1Molecular marker of (3), and identification method and application thereof |
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