CN109777886A - The molecular labeling of the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination and its application - Google Patents
The molecular labeling of the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination and its application Download PDFInfo
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Abstract
The invention discloses a kind of main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination, it is located between the label of the Indel Tal-1 on the 9th chromosome and Indel Tal-2 label, its genetic distance is 74.80cM-98.33cM, physical distance 17448080bp-22938953bp.The present invention goes back while providing the Indel label of the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination；The present invention goes back while providing the purposes of the main effect QTL: the molecular labeling by developing the main effect QTL compact linkage, to detect in rice varieties or strain whether have rice Leaf inclination correlation QTL, to accelerate the breeding process of the excellent kind of rice.
Description
Technical field
The invention belongs to rice breeding and technical field of molecular biology, and in particular to one relevant to rice Leaf inclination
Main effect QTL and its application.
Background technique
With the increase of world population quantity and the reduction of cultivated area, rice is as staple food crop in the world
One of, per unit area yield need to further increase on the basis of the original.Donald proposes the concept of ideotype at first, can be maximum
Limit improves the idealization plant type (cold language good etc., 2014) of plant photosynthesis efficiency.Hereafter, many scholars at home and abroad are to ideotype
A large amount of research is carried out, to realize the new breakthrough of rice yield.The angle that Leaf inclination, i.e. blade and stem are formed, is one
The important component of important economical character and Ideal Rice Plant Type.Song Dao saves " the ideal rice " that third (1981) propose
Morphological feature include fall three leaves want short, thick, upright, and think vertical flag leaf group than be bent group Photosynthetic Efficiency than be higher by
11%-17%.The Super-high-yielding rice strain that Yuan Longping (1997) proposes, emphasizes that three leaf of top wants straight and upright, i.e., sword-like leave, fall two leaves, fall three
The inclination angle of leaf should be respectively 5 °, 10 ° and 20 ° or so, and upright state is prolonged does not ramp up to maturation.Sun Xuchu (1985) thinks
Blade angle determine blade by light ability and to the interception of lower layer, be affected to photosynthesis.Chen Daibo etc. thinks
Three leaves the male character types in Chinese operas, usu. referring tov the bearded character degree will affect the light of rice individual and group, and then influence photosynthetic production, reasonably fall three leaves
Spatial distribution can optimize light posture, photosynthetic efficiency be improved, to improve yield.A large number of studies show that having compared with upstanding fins
Plant type can capture more luminous energy for photosynthesis, and be suitble to dense planting, lesser Leaf inclination can also improve rice leaf nitrogen product
The tired influence to kernel grouting, increasing production of rice is increased income be of great significance (Jang and Li, 2017；Olsen et al.,
2018)。
The molecular mechanism for forming Leaf inclination is different in numerous reports, but more and more researches show that, Leaf inclination
Regulation be that polygenes participates in, is related to the complex process of a plurality of signal pathway.About adjusting and controlling rice Leaf inclination dependency basis
There are many research of cause, to its molecular mechanism it is further research have larger help, however it is many research be all conceived to one or
A few gene, it is still not comprehensive for the understanding of adjusting and controlling rice Leaf inclination related gene at present.
Summary of the invention
The technical problem to be solved in the present invention is to provide the molecular labelings of the main effect QTL qTLA-9 of adjusting and controlling rice Leaf inclination
And its application.
In order to solve the above technical problem, the present invention provides a kind of main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination:
It is located between the label of the Indel Tal-1 on the 9th chromosome and Indel Tal-2 label, genetic distance 74.80cM-
98.33cM, physical distance 17448080bp-22938953bp.
Note: it falls three Leaf inclinations with more significant regulating and controlling effect to rice.
The present invention goes back while providing the Indel label of the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination, is
The primer pair of Indel Tal-1 and Indel Tal-2；
The primer pair of molecular labeling Indel Tal-1 are as follows:
Upstream primer TGGAGCACTGAATTTCTTTGGA,
Downstream primer AGCTAGTGGTAATATCTGGGTGA；
The primer pair of molecular labeling Indel Tal-2 are as follows:
Upstream primer CCAACCAGAATAAAGCCCGG,
Downstream primer GGGACCAAACCTGTTCAAGTAAG.
The present invention goes back while providing the purposes of above-mentioned main effect QTL: the molecule mark by developing the main effect QTL compact linkage
Note, to detect in rice varieties or strain whether there is rice Leaf inclination correlation QTL, to accelerate the breeding of the excellent kind of rice
Process.
The present invention goes back while providing the application of main effect QTL relevant to rice Leaf inclination: smaller for cultivating Leaf inclination
Rice optimizes plant type of rice, photosynthetic efficiency is improved, so that increasing production of rice be promoted to increase income.
The LOD value of QTL of the present invention is high；Map density is high；Mark polymorphism good.
The present invention is directed to the studies above background, and accounting for (Leaf inclination is larger)/japonica rice heat with long-grained nonglutinous rice China, to grind (Leaf inclination is smaller) miscellaneous
Handing over combined RIL group is material, has carried out rice and fall the QTL of three Leaf inclinations to position.
One of the invention is designed to provide a regulation rice and falls the main effect QTLs of three Leaf inclinations, and is named as
QTLA-9, the main effect QTL are located on the 9th chromosome, genetic distance 74.80-98.33, and physical distance is
17449080-22937953, Lod value are up to 8.53.It is right since rice Leaf inclination is one of the important indicator of Ideal Rice Plant Type
Improvement plant type of rice, promotion increasing production of rice are increased income and are of great significance, therefore, by point for developing the main effect QTL compact linkage
Son label can accelerate the choosing of the excellent kind of rice to detect in rice varieties or strain whether have rice Leaf inclination correlation QTL
Educate process.
The present invention also provides the localization methods of the main effect QTL relevant to rice Leaf inclination, comprising the following steps: 1) with
China accounts for, heat is ground and hybridized for parent, by single seed descent, obtains 134 strains for stablizing heredity, collectively constitutes RIL group；
2) recombinant inbred lines Leaf inclination Relevant phenotype data are investigated；3) it is marked using a large amount of SNP and Indel of laboratory early development
The genetic map for remembering building analyzes the label of whole chromosome group and the pass of quantitative trait phenotypes value by R-QTL professional software
QTL, is navigated to the corresponding position of linkage group, and estimate its hereditary effect by system one by one.If detecting the molecular labeling of LOD > 3,
Then think that there are 1 QTL between corresponding 2 labels of LOD value highest point.
It is a further object of the present invention to provide the molecular labelings of the main effect QTL qTLA-9 of adjusting and controlling rice Leaf inclination.
Using the primer pair of molecular labeling Indel Tal-1 and Indel Tal-2, to the DNA of rice material to be identified into
Row PCR amplification, amplified production carry out electrophoresis detection, such as amplify the DNA fragmentation of corresponding size, then indicate that adjusting and controlling rice leaf inclines
The main effect QTL qTLA-9 at angle exists.
Wherein, the primer pair of the molecular labeling Indel Tal-1, after expanding to DNA, electrophoresis can detect and water
Rice varieties heat grinds the band of same position.
Wherein, the primer pair of the molecular labeling Indel Tal-1, are as follows:
Upstream primer TGGAGCACTGAATTTCTTTGGA
Downstream primer AGCTAGTGGTAATATCTGGGTGA；
Wherein, the primer pair of the molecular labeling Indel Tal-2, after expanding to DNA, electrophoresis can detect and water
Rice varieties heat grinds the band of same position.
Wherein, the primer pair of the molecular labeling Indel Tal-2, are as follows:
Upstream primer CCAACCAGAATAAAGCCCGG
Downstream primer GGGACCAAACCTGTTCAAGTAAG.
Wherein, the reaction system of the PCR is as follows:
Wherein, PCR reaction condition are as follows: 94 DEG C of initial denaturation 3min, 94 DEG C of initial denaturation 30s, 57 DEG C of annealing 30s, 72 DEG C extend
30s expands 38 circulations, and last 72 DEG C extend 10min eventually.
It is also an object of the present invention to provide the sites main effect QTL qTLA-9 of the adjusting and controlling rice Leaf inclination
Molecule labelling method is the application that breeding is fallen in the lesser rice varieties of three Leaf inclinations.
The present invention uses the molecule labelling method of QTL site, and this method is to grind kind in rice heat using what is screened
The adjusting and controlling rice navigated on Chromosome 9 falls 2 pairs of molecular labelings of three Leaf inclination main effect QTL qTLA-9 close linkages, in advance
The Leaf inclination size for surveying rice material, accelerates the breeding of Ideal Rice Plant Type.
The present invention is ground after the first familiar generation for female parent is continuously selfed with rice variety Hua Zhanwei male parent, japonica rice variety heat and is obtained
134 recombinant inbred lines be material, three Leaf inclinations are measured and are analyzed, while having been constructed using the group
It encrypts genetic map and QTL mapping analysis is carried out to data, as a result detect that a LOD value is up to 8.53 QTL.Hair of the invention
Bright point is that the genetic linkage map spectrum density constructed is high, has 4858 labels to be uniformly distributed on 12 chromosomes, QTL is positioned more
Accurately；In addition, label polymorphism is very good, it is more convenient to Rice Offspring screening, efficient；The QTL effect value detected is larger, is
Adjusting and controlling rice falls the main effect QTLs of three Leaf inclinations, can effectively improve breeding efficiency.
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 that genetic stocks constructs flow chart in rice main effect QTL localization method relevant to Leaf inclination.
Fig. 2, which is tillering stage, falls the statistics histograms of three Leaf inclinations to plant in RIL group.
Fig. 3 is that the primer pair of molecular labeling Indel Tal-1 expands the electricity of generation in parent and its F1 generation and RIL group
Swimming figure.Wherein, 1 be China account for, 2 be heat grind, 3 be China account for/heat grind filial generation F1,4-10 be China account for/heat grinds the RIL of cross combination
Group.
Fig. 4 is that the primer pair of molecular labeling Indel Tal-2 expands the electricity of generation in parent and its F1 generation and RIL group
Swimming figure.Wherein, 1 be China account for, 2 be heat grind, 3 be China account for/heat grind filial generation F1,4-10 be China account for/heat grinds the RIL of cross combination
Group.
Fig. 5 be molecular labeling Indel Tal-1 primer pair parent 93-11, heat grind and its F1 generation and with 93-11 for wheel
Return the BC of parent3F1The electrophoretogram of generation is expanded in generation.Wherein, 1 it is 93-11,2 is that heat is ground, 3 be that 93-11/ heat grinds filial generation
F1,4-10 are using 93-11 as the BC of recurrent parent3F1The electrophoretogram of generation is expanded in generation.
Fig. 6 is that the primer pair of molecular labeling Indel Tal-2 is ground and its F1 generation and taken turns with Hua Zhanwei in parent 93-11, heat
Return the BC of parent3F1The electrophoretogram of generation is expanded in generation.Wherein, 1 it is 93-11,2 is that heat is ground, 3 be that 93-11/ heat grinds filial generation
F1,4-10 are using 93-11 as the BC of recurrent parent3F1The electrophoretogram of generation is expanded in generation.
Fig. 7 is down position of the three Leaf inclination correlation main effect QTL qTLA-9 on Chromosome 9.
Specific embodiment
The present invention is described further combined with specific embodiments below.These descriptions are not to make to the content of present invention
Further to limit, if not specified in following embodiment, technological means used is well known to those skilled in the art
Conventional means.
The acquisition of embodiment 1, experimental material
It is larger that rice varieties China accounts for Leaf inclination, and with Hua Zhanwei donor parents, local rice varieties heat is ground as receptor parent, into
Row hybridization building RILs, using single seed descent (that is, carrying out bagging single plant by kind of a processing, until offspring's strain phenotype is not sent out to F1
It is estranged from), it is final to obtain 134 strains (F13, all strain phenotypes stablize) for stablizing heredity, such as Fig. 1.
Parent and each 60 of each strain seed (F13) are chosen, is soaked seed 2 days after surface sterilization, then is wrapped up with wet towel,
It is placed in 37 DEG C of insulating boxs after vernalization 48h, selects the consistent seed sowing that shows money or valuables one carries unintentionally.After 30 days, the similar parent of upgrowth situation is selected
This and each 24 plants of transplantings of each strain rice shoot, all rice materials are planted in the biochemical academic test (examination) of Zhejiang Normal University, Jinhua, Zhejiang Province city
Test field, Routine Management.
Embodiment 2, Leaf inclination data determination
Tillering stage, every strain randomly choosed 5 tillers, measured hanging down between the vein and stem of three leaves respectively with protractor
Straight inclination angle.Each group of data is recorded, average value is calculated.
In Fig. 2, RY represents rice varieties heat and grinds, and HZ represents rice varieties China and accounts for；According to fig. 2 it can be seen that three Leaf inclination number
According to continuous normal distribution and in extensive range is shown as, with the presence of more super close individual, genetics of quantitative characters feature is shown.
Embodiment 3, QTL positioning analysis
Using the genetic map of a large amount of SNP and Indel the label building of laboratory early development, three leaves are fallen to rice and are inclined
Angle carries out quantitative trait loci (QTL) Interval mapping, and the label and quantity of whole chromosome group are analyzed by R-QTL professional software
QTL, is navigated to the corresponding position of linkage group by the relationship of trait phenotypes value one by one, and estimates its hereditary effect.If detecting LOD
> 2 molecular labeling, then it is assumed that there are 1 QTL (Fig. 3) between corresponding 2 labels of LOD value highest point.In whole chromosome group
We find a main effect QTL between the label of the Indel Tal-1 on the 9th chromosome and Indel Tal-2 label,
Genetic distance is 74.80cM-98.33cM, physical distance 17448080bp-22938953bp, and is named as qTLA-9 (figure
4)。
Embodiment 4,
Molecular labeling Indel Tal-1 and molecular labeling Indel Tal- is set separately in QTL site qTLA-9 upstream and downstream
2。
The primer pair of molecular labeling Indel Tal-1 are as follows:
Upstream primer 5 '-TGGAGCACTGAATTTCTTTGGA-3 ',
Downstream primer 5 '-AGCTAGTGGTAATATCTGGGTGA-3 '；
The primer pair of molecular labeling Indel Tal-2 are as follows:
Upstream primer 5 '-CCAACCAGAATAAAGCCCGG-3 ',
Downstream primer 5 '-GGGACCAAACCTGTTCAAGTAAG-3 '.
Take parent China to account for, heat is ground and its rice leaf of F1 generation and RIL group, extract genomic DNA, utilize above-mentioned molecule
Label carries out PCR amplification to its genomic DNA, PCR reaction system: 1 μ L of upstream primer, 1 μ L of downstream primer, 2 μ L of DNA profiling,
6 μ L of mix enzyme.Response procedures are DNA94 DEG C of initial denaturation 3min, 94 DEG C of initial denaturations 30s, 57 DEG C of annealing 30s, 72 DEG C of extension 30s, are expanded
Increase 38 circulations, last 72 DEG C extend 10min eventually.Pcr amplification product is detected in 4% agarose gel electrophoresis.
Corresponding label is determined whether there is by band analysis, heat is ground if band is intended to parent, illustrates the strain
It is smaller to be that rice falls three Leaf inclinations, illustrates if tending to China and accounting for larger.Three Leaf inclination actual corners then are fallen to tested strain rice
Degree is compared with prediction result, and prediction result matches with actually detected result as the result is shown.
The application of embodiment 5, rice Leaf inclination QTL in rice breeding
The molecular labeling set in embodiment 4 can be applied to rice molecular assistant breeding, and specific embodiment is, by other
The biggish rice varieties of Leaf inclination, such as 93-11 grind with heat and are hybridized, and obtain corresponding F1, using 93-11 as recurrent parent into
Row backcrossing, until BC3F1Generation.Extract BC3F1For part single plant DNA, then carried out with the primer of Indel label Tal-1 and Tal-2
PCR amplification determines whether there is corresponding label by band analysis, there are label explanation the strain Leaf inclination compared with
It is small.It is screened using this method, M8003 line, available Leaf inclination is smaller and remains the rice of 93-11 merit, greatly
Breeding efficiency is improved greatly.
In conclusion the three Leaf inclinations relevant main effect QTL of the invention that falls to rice can effectively reduce rice and fall three
Leaf inclination can be such that Rice Photosynthetic Efficiency is effectively promoted in breeding process, at the same can accelerate optimize rice varieties into
Journey.Leaf inclination lesser rice can be cultivated during rice molecular assistant breeding simultaneously, optimizes plant type of rice.Such side
Method is simple and easy to do, safely and effectively, is beneficial to improve the economic value of rice varieties, is suitable for large-scale promotion application.
It can be seen that the purpose of the present invention completely and is effectively achieved.Although specific embodiment party of the invention
Formula has obtained detailed description, it will be understood to those of skill in the art that.It, can be to those according to all introductions having disclosed
Details is carry out various modifications and is replaced, these changes are within the scope of the present invention.Full scope of the invention is by institute
Attached claim and its any equivalent provide.
Sequence table
<110>Zhejiang Normal University
<120>molecular labeling of the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination and its application
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Claims (4)
1. the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination, it is characterised in that: it is located at the Indel on the 9th chromosome
Tal-1 is marked between Indel Tal-2 label, genetic distance 74.80cM-98.33cM, and physical distance is
17448080bp-22938953bp。
2. the Indel of the main effect QTL site qTLA-9 of adjusting and controlling rice Leaf inclination is marked, it is characterised in that: for Indel Tal-1 and
The primer pair of Indel Tal-2；
The primer pair of molecular labeling Indel Tal-1 are as follows:
Upstream primer TGGAGCACTGAATTTCTTTGGA,
Downstream primer AGCTAGTGGTAATATCTGGGTGA；
The primer pair of molecular labeling Indel Tal-2 are as follows:
Upstream primer CCAACCAGAATAAAGCCCGG,
Downstream primer GGGACCAAACCTGTTCAAGTAAG.
3. the purposes of main effect QTL as described in claim 1, it is characterised in that: by point for developing the main effect QTL compact linkage
Son label, to detect in rice varieties or strain whether there is rice Leaf inclination correlation QTL, to accelerate the excellent kind of rice
Breeding process.
4. the application of a main effect QTL relevant to rice Leaf inclination, it is characterised in that: for cultivating the smaller rice of Leaf inclination,
Optimize plant type of rice, photosynthetic efficiency is improved, so that increasing production of rice be promoted to increase income.
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| CN111394492A (en) * | 2019-08-08 | 2020-07-10 | 中国水稻研究所 | Multi-effect QT L related to regulation of rice amylose content and gel consistency, molecular marker and application |
| CN112126703A (en) * | 2020-09-30 | 2020-12-25 | 浙江师范大学 | Molecular marker of multiple-effect QTLs locus qTLS-4 for regulating rice leaf size and application thereof |
| CN112126704A (en) * | 2020-09-30 | 2020-12-25 | 上海交通大学 | Molecular marker related to rice leaf inclination angle character and application thereof |
| CN112501341A (en) * | 2020-12-09 | 2021-03-16 | 浙江师范大学 | Major QTL for regulating heading stage of rice, molecular marker and application |
| CN112662801A (en) * | 2020-12-31 | 2021-04-16 | 浙江师范大学 | Molecular marker of major QTL site qZn1-1 for regulating and controlling zinc ion stress resistance of rice and application |
| CN113122653A (en) * | 2021-05-31 | 2021-07-16 | 浙江师范大学 | Main effect QTL for regulating and controlling brown rice rate of rice, molecular marker and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111394492A (en) * | 2019-08-08 | 2020-07-10 | 中国水稻研究所 | Multi-effect QT L related to regulation of rice amylose content and gel consistency, molecular marker and application |
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| CN112126704A (en) * | 2020-09-30 | 2020-12-25 | 上海交通大学 | Molecular marker related to rice leaf inclination angle character and application thereof |
| CN112501341A (en) * | 2020-12-09 | 2021-03-16 | 浙江师范大学 | Major QTL for regulating heading stage of rice, molecular marker and application |
| CN112662801A (en) * | 2020-12-31 | 2021-04-16 | 浙江师范大学 | Molecular marker of major QTL site qZn1-1 for regulating and controlling zinc ion stress resistance of rice and application |
| CN113122653A (en) * | 2021-05-31 | 2021-07-16 | 浙江师范大学 | Main effect QTL for regulating and controlling brown rice rate of rice, molecular marker and application |
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Application publication date: 20190521 |