CN110055348A - The Functional marker of rice grain shape gene GL3 and its application - Google Patents
The Functional marker of rice grain shape gene GL3 and its application Download PDFInfo
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- CN110055348A CN110055348A CN201910374968.3A CN201910374968A CN110055348A CN 110055348 A CN110055348 A CN 110055348A CN 201910374968 A CN201910374968 A CN 201910374968A CN 110055348 A CN110055348 A CN 110055348A
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- C—CHEMISTRY; METALLURGY
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- 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
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- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- 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
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention discloses the molecular labeling GL3-FM of rice grain shape gene Gl3 a kind of, can be used for identification and/or its offspring's assisted selection of rice grain shape improvement.Amplified fragments cannot carry the GL3-1 allele of short grain shape by the rice single plant that restriction enzyme A flIII is cut；When amplified fragments can be cut by AflIII, the plant of two band is formed, containing can result in the increased GL3-2 allele of grain length.The present invention can be used for the identification of the quick screening and GL3 allele of the long grain shape germplasm of rice, improve rice variety selective efficiency.
Description
Technical field
The invention belongs to agricultural biological technical fields, provide the molecular labeling GL3-FM of rice grain shape gene GL3 a kind of
And its application.
Background technique
Rice (Oryza sativa L.) is very important cereal crops, has more than the population of half in China with rice
Rice is staple food, and consumer groups still continuous enlargement (Zuo and Li, 2014；Yuan Longping, 2010).It is quite long in the past
In time, in order to solve the problems, such as China's short-commons, always using output increased as main target in breeding practice.It is this with height
To result in some rice varieties rice matter that China is widely applied poor for the breeding for producing as guiding.In recent years, people's living standard
Be substantially improved, requirement of the consumer to qualities such as the exterior quality of rice and boiling food flavors also increasingly improve (Mo Huidong, 1993；
Zhou Kaida etc., 1994；Fitzgerald et al,2009)；It is high-quality with the development of opening and the international trade of rice market
Rice import volume rises year by year.In the new situation, to meet the needs of internal people's level of consumption raising, the grain of country is ensured
Food safety, further increases the yield potentiality of rice, and General Promotion rice quality realizes rice high yield and good improved coordination
Have become the inexorable trend of rice breeding work.
Rice grain shape is not only directly related with rice mass of 1000 kernel, is one of the element for constituting rice yield；Particle shape still weighs
Measure Appearance Quality of Paddy Rice one of important indicator, while have an effect on rice marketing quality and processing quality (Miura et al,
2015).Grain shape is an important indicator (Kang Haiqi etc., 2001) for breeder's breeding superior rice kind.Rice grain shape is
By the complicated economical character of controlled by multiple genes, the genes for having multiple control rice grain shapes at present be cloned (Huang et al,
2013).GL3 is the gene for having cloned, having controlled rice grain length, the GL3 coding region sequence from WY3 there are two at alkali
Base makes a variation, and makes a variation at one for the C-A of 1092bp, and the C-T that another place is 1495bp makes a variation；GL3 after variation can be dramatically increased
Rice grain length (Qi et al, 2012), but variant sites are directed to, not yet develop applicable molecular labeling.
Bibliography involved in above is as follows:
1. Mo Huidong, the improvement Scientia Agricultura Sinica of China's rice quality, 1993.26 (4): p.8-14；
2. Yuan Longping, the progress China rice of super hybrid rice breeding research, 2008 (07): p.1-3；
3. week reaching, Li Hongwei, Cheng Yu, high-quality is the only way hybrid rice of hybrid paddy rice development, 1994.1994 (3-
4):p.42-46；
4.Dai,Z.,et al.,Development of a platform for breeding by design of
CMS lines based on an SSSL library in rice(Oryza sativa L.).Euphytica,
2015.205 (1): p.63-72 (Dai, Z., et al. are based on rice Single Segment Substitution Lines in Rice library construction male sterility of rice
Design of material breeding platform Euphytica, 2015.205 (1): p.63-72)；
5.Fitzgerald,M.A.,S.R.Mccouch,and R.D.Hall,Not just a grain of rice:
the quest for quality.Trends in Plant Science,2009.14(3):p.133–139
(Fitzgerald, M.A., S.R.Mccouch, and R.D.Hall, not only grain of rice, quality also extremely important Trends
in Plant Science,2009.14(3):p.133–139)；
6.Huang,R.,et al.,Genetic bases of rice grain shape:so many genes,so
Little known.Trends Plant Sci, 2013.18 (4): p.218-26 (Huang, R., et al., Genetic rice
The hereditary basis of particle shape: there are many gene for controlling particle shape, but known limited, 2013.18 (4): p.218-26)；
7.Miura,K.and M.Matsuoka,Rice genetics:Control of grain length and
Quality.Nature Plants, 2015.1 (8): p.15112 (Miura, K.and M.Matsuoka, the heredity of rice: grain
Long and quality regulation Nature Plants, 2015.1 (8): p.15112)；
8.Qi,P.,et al.,The novel quantitative trait locus GL3.1controls rice
grain size and yield by regulating Cyclin-T1；3.Cell Research,2012.22(12):
P.1666-1680 (Qi, P., et al., GL3.1, one by Cyclin T1: 3 regulate and control, control rice grain shapes and production
The gene .Cell Research of amount, 2012.22 (12): p.1666-1680)；
9.Zuo,J.and J.Li,Molecular dissection of complex agronomic traits of
rice:a team effort by Chinese scientists in recent years.National Science
Review, 2013.1 (2): p.253-276 (Zuo, J.and J.Li, rice complexity economical character molecular mechanism detailed annotation: in recent years
Chinese Scientists group work is looked back).
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of molecular labeling related with rice grain shape gene GL3 and its answer
With.
In order to solve the above technical problem, the present invention provides the molecular labelings of rice grain shape (grain length) gene GL3 a kind of
GL3-FM, using rice as species, the molecular labeling primer is selected from following primer pair, and nucleotides sequence therein is classified as 5 ' →
3 ',
GL3-FM is positive: CCCCACTGCCACATGATATGC
It is reversed: AGGCAATTGCTCCATCTCACC
Show themselves in that the amplified production of allele GL3-1 has SEQ ID when carrying out PCR amplification using the primer pair
The amplified production of nucleotide sequence shown in NO:1, allele GL3-2 has nucleotide sequence shown in SEQ ID NO:2；It is described
Allele GL3-1 shows as that grain length is shorter, and the nucleotides sequence of the allele HMA3-2 is classified as shown in SEQ ID NO:2,
Show as grain length increase.
The present invention also provides the purposes of above-mentioned molecular labeling GL3-FM: for rice grain shape improvement identification and/or its
Offspring's assisted selection.
The present invention also provides the purposes of above-mentioned molecular labeling GL3-FM:
Amplified fragments cannot carry the GL3-1 equipotential of short grain shape by the rice single plant that restriction enzyme A flIII is cut
Gene；
When amplified fragments can be cut by AflIII, the plant of two band of formation is increased containing can result in grain length
GL3-2 allele；
To realize in conjunction with breeding objective, suitable allele is selected to cultivate for rice varieties.
The improvement of purposes as molecular labeling GL3-FM of the invention:
What only length was that the plant of 688bp band carries is GL3-1 allele；
With two band, length is respectively that the plant of 408bp and 280bp carries GL3-2 allele.
The further improvement of purposes as molecular labeling GL3-FM of the invention:
PCR reaction system are as follows: 20ng/ul oryza sativa genomic dna 1ul, 10 × PCR Buffer 2.0ul, 25mM
MgCl22.0ul, 2mM dNTP 2.0ul, 10uM primer 2 .0ul, 5U/ul Taq archaeal dna polymerase 0.2ul, ddH2O
10.8ul, total system 20ul；Response procedures: 95 DEG C are denaturalized 3 minutes；94 DEG C are denaturalized 1 minute, and 55 DEG C are annealed 1 minute, 72 DEG C of extensions
1 minute, 36 circulations；72 DEG C polishing 10 minutes；
Endonuclease reaction system are as follows: the PCR product of 5ul, CutSmart Buffer 1.0ul, AflII restriction endonuclease 0.2ul,
ddH2O 3.8ul, total system 10ul；Digestion at least 3 hours in 37 DEG C of insulating boxs.
GL3 is gene of the scientific research personnel in a control rice grain length of No. 3 genomic clones of rice, according to the gene
The sequence variations of code area divide into the GL3-1 of short grain shape and the GL3-2 of long grain shape.The resulting molecular labeling GL3- of the present invention
FM is the genetic marker that rice grain length controls gene GL3, can distinguish the sequence difference of allele GL3-1 and GL3-2, Ke Yiyong
Identification and/or offspring's assisted selection in rice grain shape improve efficiency of selection.
The method provided by the invention for distinguishing allele GL3-1 and GL3-2 using above-mentioned molecular labeling GL3-FM, utilizes
Primer pair noted earlier oryza sativa genomic dna to be identified carries out PCR amplification；To PCR product AflIII restriction enzyme
Digestion, agarose electrophoresis detects the amplified production after digestion, if product cannot be cut by AflIII, agarose electrophoresis detection display
For a band, then rice to be identified has allele GL3-1；If product can be cut agarose electrophoresis inspection by AflIII
Survey is shown as two band, then rice to be identified has allele GL3-2, if agarose electrophoresis detection is shown as three rules
Band, then the GL3-2 gene of rice to be identified is heterozygous state.
Further, if agarose electrophoresis detection is shown as a band, sequence length 688bp, then rice to be identified
With allele GL3-1, it is partially short to show as grain length；If agarose electrophoresis detection is shown as two band, sequence length difference
For 408bp, 280bp, then rice to be identified has allele GL3-2, shows as grain length increase, if agarose electrophoresis detection is aobvious
Three band are shown as, sequence length is respectively 688bp, 408bp and 280bp, then the GL3 gene of rice to be identified is heterozygosis shape
State.
Using the genomic DNA of rice varieties China round-grained rice Xian 74 as template, sequence (SEQ ID that GL3-FM primer amplification obtains
NO:1):
CCCCACTGCCACATGATATGCAACTTGGATGCCACATGATATATATTATAGTTGGCATACTTTTCGTTT
GGATGAAAACTTTTAGTGGACTGGAAATTGTCAAGTTTGACATTTGTTCTCACCGTTGGATTTTATGCACCTTCAAT
TTGCTTTCTAACATCAAACGACTGCCATTGTAGGGGTACTGCTAGATGATCTTCTTGTAGCTGAAGATCTTGCTGCT
GCTGAAACGACAACCGCTGCTAATCATGCAGCGGCAAGTGCAGCAGCCACTAATGTACAAAGCGGAAGAACACCCGG
AAGATATGCTTATAATGATGAGCGAGCGAGACAAACAGCTCCAGAATCAGCTCAGGATGGATCTGTAGTTCTTGGAA
CTCCAGTTGCCCCTCCTGTCAACGGTGACATGCACACTGATATCAGCCCTGAGAATGCTGTGCTTCAGGGACAGAGG
TTATTTTCAATTTTTTTTCCCTATGAATATGTCCTGGTGATATTCCTATGGCAAGTTACTATTAGTTTTGTAATTGC
TATTTTCCTAAGGTTTCAATATTAATTTCTCTACAGGAGATTAAGCAAAGGTGTCGATTACTTAGTTGAAGCATCAG
CAGCAGAGGCAGAGGCTATTAGTGCAACTTTAGCTGCAGTGAAGGCTAGGCAGGTTAACGGTGAGATGGAGCAATTG
CCT
Using the genomic DNA of rice varieties Star bonnet 99 as template, sequence that GL3-FM primer amplification obtains
(SEQ ID NO:2):
CCCCACTGCCACATGATATGCAACTTGGATGCCACATGATATATATTATAGTTGGCATACTTTTCGTT
TGGATGAAAACTTTTAGTGGACTGGAAATTGTCAAGTTTGACATTTGTTCTCACCGTTGGATTTTATGCACCTTCA
ATTTGCTTTCTAACATCAAACGACTGCCATTGTAGGGGTACTGCTAGATGATCTTCTTGTAGCTGAAGATCTTGCT
GCTGCTGAAACGACAACCGCTGCTAATCATGCAGCGGCAAGTGCAGCAGCCACTAATGTACAAAGCGGAAGAACAC
CCGGAAGATATGCTTATAATGATGAGCGAGCGAGACAAACAGCTCCAGAATCAGCTCAGGATGGATCTGTAGTTCT
TGGAACTCCAGTTGCCCCTCCTGTCAACGGTGACATGTACACTGATATCAGCCCTGAGAATGCTGTGCTTCAGGGA
CAGAGGTTATTTTCAATTTTTTTTCCCTATGAATATGTCCTGGTGATATTCCTATGGCAAGTTACTATTAGTTTTG
TAATTGCTATTTTCCTAAGGTTTCAATATTAATTTCTCTACAGGAGATTAAGCAAAGGTGTCGATTACTTAGTTGA
AGCATCAGCAGCAGAGGCAGAGGCTATTAGTGCAACTTTAGCTGCAGTGAAGGCTAGGCAGGTTAACGGTGAGATG
GAGCAATTGCCT。
The present invention also simultaneously provide the purposes of above-mentioned molecular labeling GL3-FM: for rice difference particle shape identification with/
Or its offspring's assisted selection.
The development approach of molecular labeling of the invention, comprising the following steps:
1) led to using rice variety China round-grained rice Xian 74 as the long grain rice Star bonnet 99 of receptor parent as donor parents
Hybridization backcrossing and selfing are crossed, the increased rice Single Segment Substitution Lines in Rice material of grain length for carrying GL3-2 for background with magnificent round-grained rice Xian 74 is obtained
Material；
2) CTAB (cetyltriethylammonium bromide, Hexadecyl trimethyl ammonium Bromide) method is used
Extract rice leaf genomic DNA；
3) screening of paddy gene label is carried out using Caps molecule labelling method；
4) a Caps molecular labeling GL3-FM is developed.
With the long related molecular labeling GL3-FM of rice grain, specifically obtained with following methods:
1) according to the nucleotide sequence of gene GL3 and function variant sites, designs Caps molecular labeling, for detecting
Polymorphism between long grain shape rice Single Segment Substitution Lines in Rice material and receptor parent China round-grained rice Xian 74；
It should be noted that restriction enzyme A flIII is not the core being widely used in plant genetic engineering operation
Sour restriction endonuclease, the present invention pass through to the sequence from magnificent round-grained rice Xian 74 and from the sequence system of rice varieties Star bonnet 99
System analysis and compare, has finally been determined that AflIII is capable of the extension segment of both selective enzyme restrictions；
2) the rice Single Segment Substitution Lines in Rice material that parental rice blade and donor are Star bonnet 99 is extracted with CTAB method
Expect the genomic DNA of blade；
3) Caps molecule labelling method is used, a Caps molecular labeling GL3-FM is identified, through polymorphic detection, discovery
Itself and rice grain shape close association.
It is using the specific method that Caps molecular labeling GL3-FM carries out different rice grain length screenings:
(1) GL3-FM label different grain lengths rice Single Segment Substitution Lines in Rice material and receptor parent China round-grained rice Xian also between
DNA polymorphism analysis:
According to the nucleotide sequence of gene GL3, design develops Caps molecular labeling GL3-FM.Primer entrusts the raw work in Shanghai
Company's synthesis wins in east and carries out PCR amplification, PCR reaction system in imperial PCR instrument are as follows: 20ng/ul oryza sativa genomic dna 1ul, 10
× PCR Buffer 2.0ul, 25mM MgCl22.0ul, 2mM dNTP 2.0ul, 10uM primer 2 .0ul, 5U/ul Taq
Archaeal dna polymerase 0.2ul, ddH2O 10.8ul, total system 20ul.Response procedures: 95 DEG C are denaturalized 3 minutes；94 DEG C are denaturalized 1 minute,
55 DEG C are annealed 1 minute, and 72 DEG C extend 1 minute, 36 circulations；72 DEG C polishing 10 minutes.
The digestion of PCR product: the restriction enzyme A flIII for digestion is bought from NEB company, endonuclease reaction system
Are as follows: 5ul above-mentioned PCR product, CutSmart Buffer 1.0ul, AflII restriction endonuclease 0.2ul, ddH2O 3.8ul, total system
10ul；Digestion 3 hours or more in 37 DEG C of insulating boxs.
Digestion products detection: it is observed simultaneously under 2% agarose gel electrophoresis containing 0.5%ug/ul EB, ultraviolet lamp
Film recording result.
(2) it is educated using the screening of the Caps label GL3-FM progress long particle shape germ plasm resource of rice and the assisted Selection of particle shape
Kind:
Rice grain shape is the important target character in breeding selection.Particle shape is the complicated economical character by controlled by multiple genes,
It is difficult the allelic gene type that intuitive judgment from the appearance goes out related particle shape gene in rice material.By will long grain Star
Bonnet 99 and magnificent 74 hybridization of round-grained rice Xian, continuous backcross binding marker assisted Selection, imported into magnificent round-grained rice Xian for long grain gene GL3-2
In 74, GL3-FM banding pattern and the consistent material of Star bonnet 99 are selected, the seed tied on these plant is harvested, detects it
Grain length finds that its grain length is significantly improved compared with receptor parent China round-grained rice Xian 74.
Molecular labeling GL3-FM can be used for Rice Germplasm Resources screening.In the present invention, when rice plant to be identified passes through
When 688bp band occurs in detection, we determine that it carries the GL3-1 allele of short grain shape；When plant to be identified is through detecting
When there is 408bp and two band of 280bp, we determine that it carries the GL3-2 allele of long grain shape.The present invention is applicable in all
Identify and screen based on the allele that C-T on the site GL3 gene 1495bp makes a variation.
The present invention also provides purposes of the molecular labeling GL3-FM noted earlier in identification rice grain length character.
The present invention also provides the methods noted earlier for distinguishing allele GL3-1 and GL3-2 to educate in rice grain shape improvement
Purposes in kind.
The present invention has following technical advantage:
(1) present invention is studied by influencing the natural variation of the major gene resistance GL3 of rice grain length on control, develops one
The molecule mark on the gene coding region of short grain allelotype GL3-1 and long grain allelotype GL3-2 can rapidly and efficiently be distinguished
Remember GL3-FM.
(2) present invention be based on rice grain length gene GL3 nucleotide sequence development and obtain Caps molecular labeling,
The label is located at the gene coding region GL3, can effectively identify rice grain shape character, and be stabilized, and can be rice grain shape
Design and context provides new technological means, accelerates Breeding Process.
(3) the rice Single Segment Substitution Lines in Rice material under magnificent 74 background of round-grained rice Xian that (embodiment 1) of the invention obtains is based on water
The breed breeding of grain of rice shape improvement provides good breeding material.
Although nucleotide variation at prior art discloses the GL3 coding region sequences from WY3 there are two, it is at one
The C-A of 1092bp makes a variation, and the C-T that another place is 1495bp makes a variation；In the embodiment 1 shown in the present invention, NIL-GL3-2 with
NIL-GL3-1 (i.e. HJX74) only has the C-T variation in the site 1495bp, and the two also has significant phenotypic difference, and the present invention also provides
One it is novel, increase the long GL3 allele of rice grain, i.e. GL3-2.Due to GL3-2 and GL3-1 have and only one
Base difference is difficult to distinguish using SSR marker etc., and the prior art is analyzed often through sequence and determines the specific base sequence in the site
Column.Specific practice are as follows: according to other adjacent sequence design gene-specific primer, to the genomic DNA fragment comprising the variant sites
PCR amplification is carried out, amplified band is checked in agarose gel electrophoresis, the DNA item that selection meets expected size is brought into
Row recycling delivers sequencing company and carries out sequence analysis.It is marked relative to Caps of the present invention, previous detection method needs
Longer detection time usually needs how time-consuming 1~2 day than digestion detection including feeding sample sequencing, at the same time, DNA
The cost of segment recycling and Sequence Detection is also much higher than Caps label, and testing cost increases 10 times or more.The present invention provides
A kind of GL3-2 allele screening technique of simple and effective.
In conclusion the present invention provides the molecular labeling of rice grain length gene GL3 a kind of and its applications.The present invention is directed to
The design primer in the site for causing particle shape to make a variation, expands oryza sativa genomic dna using PCR system, then produces to amplification
Object carries out digestion, determines the GL3 allele contained in rice to be measured finally by agarose electrophoresis detection.The present invention can use
In the identification of the quick screening and GL3 allele of the long grain shape germplasm of rice, rice variety selective efficiency is improved.That is, the molecule
Label can effectively identify the GL3 allele in rice.In breeding process, by identifying, excellent genetic resources are utilized
With corresponding molecular marker assisted selection means, efficiency of selection can greatly be improved, helps to carry out using the gene loci
Molecular design breeding.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing.
Fig. 1 is the rice Single Segment Substitution Lines in Rice material under magnificent 74 background of round-grained rice Xian identified using molecular labeling GL3-FM
Phenotypic map；
A. the NIL-GL3-1 plant type of GL3-1 allele is carried, NIL-GL3-1 is to construct rice Single Segment Substitution Lines in Rice
Receptor parent China round-grained rice Xian 74, bar=10cm；
B. the NIL-GL3-2 plant type of GL3-2 allele, bar=10cm are carried；
The particle shape of c.NIL-GL3-1 and NIL-GL3-2, bar=0.5cm.
Fig. 2 is the electrophoretic band figure that rice varieties GL3 gene type is identified using molecular labeling GL3-FM；
M representation DNA molecular weight Marker, 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,
20,21,22,23,24,25,26,27 represent Different Rice Varieties, be followed successively by Tetep, Amol3, in 4188, BG 367, Zihui
100, Katy, Suyunuo, IR 64, Basmati 385, Nan Yang account for, Basmati 370, IR58025B, Jiangxi silk seedling, join mirror
33, U.S. jasmine perfume (or spice) rice, Jiangxi perfume (or spice) is glutinous, IRAT 261, KYEEMA, Cheng Long's crystal rice, IR65598-112-2, Khazar,
Lemont, Star bonnet 99, IAPAR9, IR66897B, IR66167-27-5-1-6 and magnificent round-grained rice Xian 74.
Fig. 3 is the phenotype using the particle shape gene pyramiding based material of molecular labeling GL3-FM breeding.
A. the grain of rice material is long, and the grain of b. rice material is wide, the mass of 1000 kernel of c. rice material.
Specific embodiment
Embodiment 1. marks GL3-FM assisted Selection with Caps, screens under magnificent 74 background of round-grained rice Xian, carries the single slice of GL3-2
Replace based material.
Specifically: it with rice varieties China round-grained rice Xian 74 is receptor parent, Star bonnet 99 is donor parents, passes through routine
Hybridization and backcrossing, construct a series of rice Single Segment Substitution Lines in Rice materials.It is female parent with magnificent round-grained rice Xian 74, Star bonnet 99 is
Male parent carries out conventional hybridization, obtains F1For cenospecies, F is obtained after selfing is homozygous2For material, then to the F of acquisition2It is random for material
Menu strain is returned using magnificent round-grained rice Xian 74, and after continuous backcross at least 3 times, selfing is homozygous.To the homozygous material use molecule mark of acquisition
Remember assisted Selection, determine the chromosome segment from Star bonnet 99 carried in different single plants, by identification, has and only
There is the chromosome segment material from Star bonnet 99 to be defined as Single Segment Substitution Lines in Rice material.It is auxiliary by molecular labeling
Selection is helped, the material (NIL-GL3-2) for carrying GL3-2 allele is identified using GL3-FM, which significantly increases
(Fig. 1).
1.DNA is extracted
According to requirement of experiment, the extracting of DNA uses TPS simplified method.TPS simplified method: (1) it in tillering regularity each single plant takes
Top 1-2 piece spire, is stored in spare in -80 DEG C of refrigerators；(2) rice leaf of 2-4cm long is taken to be put into 2.0mL when extracting DNA
In centrifuge tube, it is put into liquid nitrogen, with grinding rod grinds；(3) plus TPS extract 800 μ L, 75 DEG C water-bath 30-60 minutes；(4) high
Fast refrigerated centrifuge 12000rpm is centrifuged 10 minutes.It sucts about 500 μ L of clear liquid and is transferred to 1.5mL centrifuge tube；(5) it is added isometric
The isopropanol or dehydrated alcohol of pre-cooling are placed to DNA at 4 DEG C and are precipitated；(6) high speed low temperature centrifugal machine 12000rpm is centrifuged 10 points
Clock abandons supernatant；(5) naturally dry, is added 200 μ L sterilizing distilled water dissolution, and 4 DEG C of preservations are placed.
TPS extract ingredient: 100mM Tris-HCl (pH8.0), 10mM EDTA (pH8.0), 1M KCl.
2.PCR amplification
1) reaction system:
Using the genomic DNA of extraction as template, GL3-FM is that primer carries out PCR reaction.Reaction system includes: paddy gene
Group DNA 20ng/ul 1ul, 10 × PCR Buffer 2.0ul, 25mM MgCl22.0ul, 2mM dNTP 2.0ul, 10uM draw
Object each 1.0ul, 5U/ul Taq archaeal dna polymerase 0.2ul, ddH2O 10.8ul, total system 20ul.
2) response procedures:
95 DEG C are denaturalized 3 minutes；94 DEG C are denaturalized 30 seconds, and 55 DEG C are annealed 30 seconds, and 72 DEG C extend 1 minute, 36 circulations；72 DEG C neat
10 minutes, 12 DEG C 5 minutes.
The digestion system of 3.PCR amplified production
Take above-mentioned PCR product 5.0ul, ddH2O 3.8ul, 10 × NEB CutSmart Buffer 1.0ul, restricted core
Sour restriction endonuclease AflIII 0.2ul, in total 10.0ul, 37 DEG C constant temperature digestion 3 hours or more.
4. agarose gel electrophoresis detects
Glue: the preparation method of 2% Ago-Gel: weighing 1.6g agarose, and addition fills 1 × tbe buffer of 80mL
In the triangular flask of liquid, it is heated to agarose and is completely dissolved.After being cooled to about 60 DEG C, the 50 μ L of EB of 1mg/mL is instilled, is shaken up.It will
Agarose solution pours into plastic plate, plugs comb and drives bubble away.
Point sample: 3 μ L bromophenol blue indicators are added in product after the resulting PCR digestion of above-mentioned steps 3, take the sample of 7.5 μ L
Product are imaged with gel prime minister's system after electrophoresis 30min under 130V voltage and record result.
According to electrophoresis result, what an only length was that the plant of 688bp band carries is GL3-1 allele；Have
Two band, length are respectively that the plant of 408bp and 280bp carries GL3-2 allele.
It selects to carry the plant of GL3-2 allele, i.e. near isogenic lines NIL-GL3-2 under magnificent 74 background of round-grained rice Xian.The material
Material in plant type with magnificent round-grained rice Xian 74 closely similar (Fig. 1 a, b).
Magnificent round-grained rice Xian 74 (NIL-GL3-1) and NIL-GL3-2 kind are planted until harvest rice grain；The particle shape of NIL-GL3-2
Obvious with magnificent round-grained rice Xian 74 (NIL-3-1) difference, seed length dramatically increases (Fig. 1 c).GL3-FM can be used as identification grain length
Functional label uses.The increased Single Segment Substitution Lines in Rice material NIL-GL3-2 of the grain length that the present embodiment screens can also be used as rice
The molecular breeding design element of particle shape improvement.
Embodiment 2. carries the Rice Germplasm Resources of GL3-2 using molecular labeling GL3-FM identification
Specific practice is: Rice Germplasm Resources of the random selection from multiple countries and regions in the world extract blade base
Because of a group DNA, the allelic gene type of these genetic stocks is identified using GL3-FM.
1.DNA is extracted
With embodiment 1.
2.PCR amplification
With embodiment 1.
The digestion system of 3.PCR amplified production
With embodiment 1.
4. agarose gel electrophoresis detects
With embodiment 1.
According to fig. 2, it can be concluded that GL3-FM can be used for identifying the GL3-FM equipotential base in different rice materials
Because of type.As shown in Figure 2, electrophoresis result only has the material of a band to carry GL3-1 allele, and banding pattern pronounces " 1 "；Have
The material of two band carries GL3-2 allele, and banding pattern pronounces " 3 ".Wherein, for constructing Single Segment Substitution Lines in Rice and exploitation mark
The number of material China round-grained rice Xian 74 of note is 27, and an only band carries GL3-1 allele；Star as donor parents
99 electrophoresis result of bonnet is two bands, is carried GL3-2 allele (table 1).
Table 1, the rice material for identification and GL3 genotype
GL3-FM banding pattern is the kind of " 1 " in above-mentioned such as table 1, after measured, containing GL3-1 described in SEQ ID NO:1
Allele；GL3-FM banding pattern is the kind of " 3 " in above-mentioned such as table 1, after measured, containing sequence described in SEQ ID NO:2
GL3-2 allele.
Illustrate: for resource material listed in table 1, although what is had contains GL3-2, grain length is also partially short, this is being cultivated
It is normal in material, is the result that many genes concur because different materials determine that the factor of particle shape is very more.This
The genotype that can be used for identifying GL3 in material is only verified in invention, regardless of phenotype.
Embodiment 3. carries out the assisted Selection and the building of polymerization system of particle shape gene using molecular labeling GL3-FM
Using under 74 background of round-grained rice Xian have been built up, magnificent, list of the GW5 allele from Large grain rice kind Suyunuo
Segment materials (are named as NIL-GW5), and material use Large grain rice kind Suyunuo is miscellaneous with magnificent round-grained rice Xian 74 as donor parents
It hands over, single plant and magnificent round-grained rice Xian 74 continuous backcrosses at least 4 generations is selected in offspring, it is homozygous to eventually pass through selfing.In the material of acquisition
It is screened using GW5 specific marker, obtains NIL-GW5.For the particle shape of NIL-GW5 compared with magnificent round-grained rice Xian 74, rice grain is long
It reduces, but grain width dramatically increases.
The quotation of GW5: Shomura A, et al.Deletion in a gene associated with grain
Size increased yields during rice domestication.Nat Genet, 2008,40:1023-1028
(Shomura A, et al. rice evolve in the particle shape as caused by sequence deletion change and output increased .Nat Genet, 2008,
40:1023-1028).
The material is hybridized with NIL-GL3-2 described in embodiment 1, construct while carrying GW5 allele and GL3-2
The dual-gene polymerization based material NIL-GL3-2/GW5 of allele.Specific practice is: selecting to carry grain under magnificent 74 background of round-grained rice Xian
The Single Segment Substitution Lines in Rice material of wide gene GW5, is hybridized with NIL-GL3-2, combines GL3-FM molecule mark to the offspring of acquisition
Row assisted Selection is remembered into, in F1In generation, selects true cenospecies, plantation at F2Group, in F2In segregating population further using mark into
Row identification, screens the homozygous dual-gene polymerization based material of GW5/GL3-2, and the design and context for rice grain shape is improved.That is, utilizing
Molecular marker screening improves the efficiency for obtaining gene pyramiding system, reaches and increases mass of 1000 kernel, improves particle shape, and then it is excellent to reach high yield
The purpose of matter improved coordination.
It is specific as follows:
1.DNA is extracted
With embodiment 1.
2.PCR amplification
With embodiment 1.
The digestion system of 3.PCR amplified production
With embodiment 1.
4. agarose gel electrophoresis detects
With embodiment 1.
According to electrophoresis result, what an only length was that the plant of 688bp band carries is GL3-1 allele；Have
Two band, length are respectively that the plant of 408bp and 280bp carries GL3-2 allele.Detection for GW5 allele
Using the primer of Shomura (2008) et al., when obtaining one about length being the DNA band of 1.5Kb, show that material carries
Gw5 allele from magnificent round-grained rice Xian 74；The material of amplified band about 300bp carries the GW5 allele from Suyunuo.?
F2In group, the NIL- of screening while the as quasi- building of the material for carrying homozygous GL3-2 allele and homozygosis GW5 allele
The dual-gene polymerization based material of GL3-2/GW5.
5. the investigation of rice grain shape character
After grain complete ripeness, by single plant sowing, full maturity is chosen, full grain is enriched and investigates character.
Grain is long: each single plant takes morphologically normal 30 grain to be averaged with every grain length of vernier caliper measurement at random
Value；
Grain is wide: each single plant takes morphologically normal 30 grain at random, wide with every grain of vernier caliper measurement, is averaged
Value；
Mass of 1000 kernel: grain is sufficiently dried, and removes blighted grain, is taken at random and is enriched good seed 1,000 weighing and (or take
500 weights are converted into mass of 1000 kernel again), the difference of weight twice of being subject to is not more than the 3% of its average value, otherwise reforms, is with gram
Unit.
According to Fig. 3, NIL-GW5 and donor parents (magnificent round-grained rice Xian 74, be abbreviated as HJX4) are compared, and the length of grain reduces, paddy
Grain is wide to be dramatically increased, and mass of 1000 kernel is also improved；
Compared with donor parents (magnificent round-grained rice Xian 74, be abbreviated as HJX4), NIL-GL3-2 grain length dramatically increases, grain it is wide with
The grain width of magnificent round-grained rice Xian 74 is similar, and mass of 1000 kernel is also above magnificent round-grained rice Xian 74.
After two gene pyramidings, obtain that grain is long, the wide material improved of grain, especially mass of 1000 kernel, not only far
Higher than the mass of 1000 kernel of magnificent round-grained rice Xian 74, the mass of 1000 kernel also than NIL-GW5 and NIL-GL3-2 for polymerization is high (Fig. 3).The above table
Bright, GL3-FM can be used in the design and context of rice grain shape improvement, greatly improve efficiency of selection.
The above list is only a few specific embodiments of the present invention for finally, it should also be noted that.Obviously, this hair
Bright to be not limited to above embodiments, acceptable there are many deformations.Those skilled in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Sequence table
<110>Agricultural University Of South China
<120>Functional marker of rice grain shape gene GL3 and its application
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 688
<212> DNA
<213>rice (Oryza sativa)
<400> 1
ccccactgcc acatgatatg caacttggat gccacatgat atatattata gttggcatac 60
ttttcgtttg gatgaaaact tttagtggac tggaaattgt caagtttgac atttgttctc 120
accgttggat tttatgcacc ttcaatttgc tttctaacat caaacgactg ccattgtagg 180
ggtactgcta gatgatcttc ttgtagctga agatcttgct gctgctgaaa cgacaaccgc 240
tgctaatcat gcagcggcaa gtgcagcagc cactaatgta caaagcggaa gaacacccgg 300
aagatatgct tataatgatg agcgagcgag acaaacagct ccagaatcag ctcaggatgg 360
atctgtagtt cttggaactc cagttgcccc tcctgtcaac ggtgacatgc acactgatat 420
cagccctgag aatgctgtgc ttcagggaca gaggttattt tcaatttttt ttccctatga 480
atatgtcctg gtgatattcc tatggcaagt tactattagt tttgtaattg ctattttcct 540
aaggtttcaa tattaatttc tctacaggag attaagcaaa ggtgtcgatt acttagttga 600
agcatcagca gcagaggcag aggctattag tgcaacttta gctgcagtga aggctaggca 660
ggttaacggt gagatggagc aattgcct 688
<210> 2
<211> 688
<212> DNA
<213>rice (Oryza sativa)
<400> 2
ccccactgcc acatgatatg caacttggat gccacatgat atatattata gttggcatac 60
ttttcgtttg gatgaaaact tttagtggac tggaaattgt caagtttgac atttgttctc 120
accgttggat tttatgcacc ttcaatttgc tttctaacat caaacgactg ccattgtagg 180
ggtactgcta gatgatcttc ttgtagctga agatcttgct gctgctgaaa cgacaaccgc 240
tgctaatcat gcagcggcaa gtgcagcagc cactaatgta caaagcggaa gaacacccgg 300
aagatatgct tataatgatg agcgagcgag acaaacagct ccagaatcag ctcaggatgg 360
atctgtagtt cttggaactc cagttgcccc tcctgtcaac ggtgacatgt acactgatat 420
cagccctgag aatgctgtgc ttcagggaca gaggttattt tcaatttttt ttccctatga 480
atatgtcctg gtgatattcc tatggcaagt tactattagt tttgtaattg ctattttcct 540
aaggtttcaa tattaatttc tctacaggag attaagcaaa ggtgtcgatt acttagttga 600
agcatcagca gcagaggcag aggctattag tgcaacttta gctgcagtga aggctaggca 660
ggttaacggt gagatggagc aattgcct 688
Claims (5)
1. the molecular labeling GL3-FM of rice grain shape gene Gl3, using rice as species, it is characterized in that: the molecular labeling draws
Object is selected from following primer pair, and nucleotides sequence therein is classified as 5 ' → 3 ',
GL3-FM is positive: CCCCACTGCCACATGATATGC
It is reversed: AGGCAATTGCTCCATCTCACC.
2. the purposes of molecular labeling GL3-FM as described in claim 1, it is characterized in that: the identification for rice grain shape improvement
And/or its offspring's assisted selection.
3. the purposes of molecular labeling GL3-FM as described in claim 1, it is characterized in that:
Amplified fragments cannot carry the GL3-1 allele of short grain shape by the rice single plant that restriction enzyme A flIII is cut；
When amplified fragments can be cut by AflIII, the plant of two band is formed, containing can result in the increased GL3-2 of grain length
Allele.
4. the purposes of molecular labeling GL3-FM according to claim 3, it is characterized in that:
What only length was that the plant of 688bp band carries is GL3-1 allele；
With two band, length is respectively that the plant of 408bp and 280bp carries GL3-2 allele.
5. the purposes of molecular labeling GL3-FM according to claim 3 or 4, it is characterized in that:
PCR reaction system are as follows: 20ng/ul oryza sativa genomic dna 1ul, 10 × PCR Buffer 2.0ul, 25mM MgCl2
2.0ul, 2mM dNTP 2.0ul, 10uM primer 2 .0ul, 5U/ul Taq archaeal dna polymerase 0.2ul, ddH2O 10.8ul, it is overall
It is 20ul；Response procedures: 95 DEG C are denaturalized 3 minutes；94 DEG C are denaturalized 1 minute, and 55 DEG C are annealed 1 minute, and 72 DEG C extend 1 minute, 36
Circulation；72 DEG C polishing 10 minutes；
Endonuclease reaction system are as follows: the PCR product of 5ul, CutSmart Buffer 1.0ul, AflII restriction endonuclease 0.2ul, ddH2O
3.8ul, total system 10ul；Digestion at least 3 hours in 37 DEG C of insulating boxs.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110592264A (en) * | 2019-10-17 | 2019-12-20 | 青岛农业大学 | Molecular marking method of peanut plant type related gene locus and application thereof |
| CN111218445A (en) * | 2020-02-10 | 2020-06-02 | 扬州大学 | Method for improving quality of vertical ear type japonica rice and molecular marker |
| CN114164291A (en) * | 2021-11-10 | 2022-03-11 | 华南农业大学 | Application of rice grain length gene GL10 allele |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103289972A (en) * | 2012-03-02 | 2013-09-11 | 中国科学院上海生命科学研究院 | Rice long-grain related gene and application for same |
| CN103866036A (en) * | 2014-04-02 | 2014-06-18 | 江苏省农业科学院 | Method for identifying genes of 1000-grain weight of rice TGW6 by molecular markers |
| CN103374578B (en) * | 2012-04-16 | 2014-11-12 | 华中农业大学 | Gene G13 for regulating length and weight of rice grains and application |
| CN104450897A (en) * | 2014-11-24 | 2015-03-25 | 中国水稻研究所 | Functional specificity molecular marker for rice high yield allele GY6 |
| CN105585619A (en) * | 2014-11-12 | 2016-05-18 | 中国农业大学 | Protein related to grain length and grain weight of paddy rice grains, encoding gene GL3-3 and application thereof |
| CN106521021A (en) * | 2017-01-05 | 2017-03-22 | 江西省农业科学院水稻研究所 | Gene marker for identifying haplotypes of rice grain width and grain weight GS5 gene and application |
| WO2017106274A1 (en) * | 2015-12-16 | 2017-06-22 | Syngenta Participations Ag | Genetic regions & genes associated with increased yield in plants |
-
2019
- 2019-05-07 CN CN201910374968.3A patent/CN110055348A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103289972A (en) * | 2012-03-02 | 2013-09-11 | 中国科学院上海生命科学研究院 | Rice long-grain related gene and application for same |
| CN103374578B (en) * | 2012-04-16 | 2014-11-12 | 华中农业大学 | Gene G13 for regulating length and weight of rice grains and application |
| CN103866036A (en) * | 2014-04-02 | 2014-06-18 | 江苏省农业科学院 | Method for identifying genes of 1000-grain weight of rice TGW6 by molecular markers |
| CN105585619A (en) * | 2014-11-12 | 2016-05-18 | 中国农业大学 | Protein related to grain length and grain weight of paddy rice grains, encoding gene GL3-3 and application thereof |
| CN104450897A (en) * | 2014-11-24 | 2015-03-25 | 中国水稻研究所 | Functional specificity molecular marker for rice high yield allele GY6 |
| WO2017106274A1 (en) * | 2015-12-16 | 2017-06-22 | Syngenta Participations Ag | Genetic regions & genes associated with increased yield in plants |
| CN108697752A (en) * | 2015-12-16 | 2018-10-23 | 先正达参股股份有限公司 | With the relevant genetic region of the increased yield of plant and gene |
| CN106521021A (en) * | 2017-01-05 | 2017-03-22 | 江西省农业科学院水稻研究所 | Gene marker for identifying haplotypes of rice grain width and grain weight GS5 gene and application |
Non-Patent Citations (6)
| Title |
|---|
| AYAHIKO SHOMURA等: "Deletion in a gene associated with grain size increased yields during rice domestication", 《NAT GENET.》 * |
| PENG QI等: "The novel quantitative trait locus GL3.1 controls rice grain size and yield by regulating Cyclin-T1;3", 《CELL RESEARCH》 * |
| 江卫平等: "水稻粒重生理特性及遗传分析研究进展", 《安徽农业科学》 * |
| 裔传灯等: "水稻粒宽基因GS5的功能标记开发和单倍型鉴定", 《中国水稻科学》 * |
| 裔传灯等: "水稻粒形基因GS3 的功能标记开发与鉴定", 《江苏农业科学》 * |
| 高虹等: "水稻粒长基因GL3 的遗传分析和分子标记定位", 《植物生理学通讯》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110592264A (en) * | 2019-10-17 | 2019-12-20 | 青岛农业大学 | Molecular marking method of peanut plant type related gene locus and application thereof |
| CN111218445A (en) * | 2020-02-10 | 2020-06-02 | 扬州大学 | Method for improving quality of vertical ear type japonica rice and molecular marker |
| CN114164291A (en) * | 2021-11-10 | 2022-03-11 | 华南农业大学 | Application of rice grain length gene GL10 allele |
| CN114164291B (en) * | 2021-11-10 | 2024-01-23 | 华南农业大学 | Application of rice grain length gene GL10 allele |
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