CN109880929A - The molecular labeling of the main effect QTL site Apam-1 of adjusting and controlling rice bacterial brown streak resistance and its application - Google Patents
The molecular labeling of the main effect QTL site Apam-1 of adjusting and controlling rice bacterial brown streak resistance and its application Download PDFInfo
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Abstract
The invention discloses a kind of main effect QTL Apam-1 of adjusting and controlling rice bacterial brown streak resistance, QTL site is located on No. 1 chromosomal dna fragment, genetic distance 162.95-172.83cM, physical distance 38013068bp-40317380bp.The present invention goes back while providing the Indel label of the main effect QTL site Apam-1 of adjusting and controlling rice bacterial brown streak resistance, is the primer pair of Bbsr-1 and Bbsr-2.The present invention goes back while providing a kind of method of artificial infection idenfication rice plant disease resistance.The present invention is screened using molecule labelling method, M8003 line, be can get the rice with cecospora spot resistance, is substantially increased breeding efficiency.
Description
Technical field
The invention belongs to paddy disease-resistant breeding and molecular biology fields, and in particular to the bacillary brown item of an adjusting and controlling rice
The molecular labeling of the main effect QTL site Apam-1 of sick resistance and its application.
Background technique
Pseudomonas panici Stapp (Bacterial brown stripe of rice, BBSR) is also known as the paddy bacterial heart
Maize ear rot is a kind of seed-borne disease that bacterial bearing rate is high.The disease is most found earlier than last century the fifties in Japan, at present in Asia
It is all had been reported that in the world wides such as continent, Africa, Europe, America, is distributed mainly on the Yangtze river basin and south China rice producing region in China
(Liu congratulates, 2015).Pseudomonas panici Stapp disease incidence when big Tanaka's shallow water manages is very low, 0.07% hereinafter, symptom master
Show as the single clump of withered heart.It can be in primary infection focus diffused if often keeping the rice field of deep water, more clumps concentrate in together morbidity,
Disease incidence is up to 1% or so.If big water logging seedling, clump disease incidence is high up to 100% all 15% or so, this production to rice
Amount and quality will cause great harm.Be often accompanied by heavy rain when in recent years, with morning, the semilate rice season of growth, rice seedling flooded or
The probability invaded by flood increases so that brown streak is locally being very popular so that Rice Yield Loss Caused seriously (Yang Chunlan,
2015).Obvious morbidity is seen in a week after general water logging.Primary diseased plant is withered cardioid, and it is first withered with side leaf to be infected strain symptom
Based on brown symptom, withered heart symptom then or simultaneously occurs.Such as high temperature fine day, the withered heart shows as the acute of green withered distortion cylinder volume
Symptom, it will thus be seen that water is the approach for infecting diffusion again；Flood overflows seedling, and wound is more, intrusion morbidity it is also more (Li Jun equality,
2002).Oat acidovorax avenae oat subspecies (Acidovorax avenae sub.avenae) are the pathogens for causing this kind of disease,
The plant infected by the bacterium is mostly just withered before heading, the normal early fringe survived.In boot stage, fringe bud, which is caught an illness, causes fringe early
It is withered, fringe neck abnormal elongation, small ear obstruct filbert and deformity, grain is brown not firm, part spike of rice or do not extend also just from
Break leaf sheath under sword-like leave pulvinus and go out, or be rolled in leaf sheath it is interior at " tire of suffocating " (Wang Li, 2016；Shen Rongping etc., 2015).
Pseudomonas panici Stapp morbidity is serious, and prevention and treatment is difficult, and the control measure taken in production at present mainly has agricultural
Prevention and treatment and chemical prevention.Although these measures play certain preventive and therapeutic effect, rice bacterium cannot be fundamentally reduced
The generation of property brown streak.Resistant gene is a variety of defense mechanisms being evolved during rice and the long-term interaction of pathogen
One of.The bacteria resistance brown streak genetic locus on rice genome is excavated, is imported into our common main cultivated rice cultivars,
To adapt to the different weather conditions in various regions, the disease incidence of brown streak can be greatly reduced, and be a kind of most safe and economical effective
Controlling way.So far, though QTL Position Research has more report on rice, in Rice Resistance bacterial brown streak gene
Report in terms of QTL positioning analysis is considerably less, passes through building genetic linkage maps and quantitative trait locus (Quantitative
Trait Locus, QTL) analysis, it can effectively find main effect QTL relevant to Pseudomonas panici Stapp resistance and apply
In rice molecular assistant breeding, rice plant is improved, cost has been saved, has also improved breeding efficiency.
Summary of the invention
The technical problem to be solved in the present invention is to provide the main effect QTL sites of adjusting and controlling rice bacterial brown streak resistance
The molecular labeling of Apam-1 and its application.
In order to solve the above technical problem, the present invention provides a kind of main effect QTLs of adjusting and controlling rice bacterial brown streak resistance
The site Apam-1:QTL is located on No. 1 chromosomal dna fragment, genetic distance 162.95-172.83cM, and physical distance is
38013068bp-40317380bp, LOD value are up to 4.67.
Note: it has stronger resistance to Pseudomonas panici Stapp.QTL refer to controlling the size character gene in gene
Position in group.
The present invention goes back while providing the Indel of the main effect QTL site Apam-1 of adjusting and controlling rice bacterial brown streak resistance
Label: for the primer pair of Bbsr-1 and Bbsr-2；
The primer pair of molecular labeling Indel Bbsr-1 are as follows:
Upstream primer 5 '-CATAGCATTTTCGATTTTTGAA-3 '
Downstream primer 5 '-CTTGTGACGGAACCAAATCA-3 '；
The primer pair of molecular labeling Indel Bbsr-2 are as follows:
Upstream primer 5 '-AATGATGAGGGGAGTGGATG-3 '
Downstream primer 5 '-AGCTACTGGTGGTGCTTGCT-3 '.
The present invention goes back while providing a kind of method of artificial infection idenfication rice plant disease resistance:
Accounted for using China, heat is ground and its progeny population is research object, artificial infection brown streak bacterium RS-1, parents China is accounted for, heat
It grinds and the plant height of each strain of group is measured and (carried out after connecing bacterium 3 weeks, measurement is specific with quantitative analysis with quantitative analysis
Content is to measure the height of rice highest point upwards from Culm of Rice bottom with meter ruler, and each strain measures 4 plants respectively and takes over bacterium
Rice and 4 plants of rice for not taking over bacterium), it is resistant to Pseudomonas panici Stapp to judge whether plant has.
Criterion are as follows: when the rice that bacterium is taken in the strain be averaged plant height/do not connect bacterium rice be averaged plant height >
When 0.96, determine that the strain is resistant to Pseudomonas panici Stapp；Conversely, when the rice for taking over bacterium in the strain is averagely planted
Plant height degree/do not connect bacterium rice be averaged plant height≤0.96 when, then it is anti-to determine that the strain does not have Pseudomonas panici Stapp
Property.
Note: it is resistance parent that heat, which is ground, and after heat grinds and connects bacterium, rice plant height is reduced to the 96% of normal plant, therefore
It selects 0.96 as the index for judging whether rice has brown streak resistance.
If plant pair brown streak be it is susceptible, the plant height of inoculated brown streak bacterium can be significantly lower than normal plant.
Tiny brown marking can all occur in entire plant when rice brown streak hair, as finally entire plant is rotted for the sprawling of germ,
It is not easy to quantify.
The molecular labeling that the present invention went back while providing the main effect QTL of an adjusting and controlling rice bacterial brown streak resistance is answered
With, screened using molecule labelling method, M8003 line, can get with cecospora spot resistance rice, substantially increase and educate
Kind efficiency.
Label polymorphism of the invention is good, and selection is convenient, efficiently.
The present invention is directed to the studies above background, accounts for the RIL group that (perception)/japonica rice heat grinds (resistance) cross combination with long-grained nonglutinous rice China
Body is material, has carried out the QTL positioning of anti-Pseudomonas panici Stapp character.
One of the objects of the present invention is to provide the main effect QTL sites of an adjusting and controlling rice bacterial brown streak resistance, and
Be named as Apam-1, the main effect QTL is located on No. 1 chromosomal dna fragment, Indel mark Bbsr-1 and
Indel is marked between Bbsr-2；Genetic distance is 162.95-172.83cM, physical distance 38013068bp-
40317380bp, LOD value are up to 4.67, have significant Pseudomonas panici Stapp resistance, can reduce in breeding process
It even is eliminated harm of the Pseudomonas panici Stapp to rice plant.Therefore, the present invention also protects the adjusting and controlling rice bacterium
Property brown streak resistance main effect QTL optimization rice varieties in application.
The present invention also provides the localization method of the main effect QTL of the adjusting and controlling rice bacterial brown streak resistance, including it is following
Step: 1) being accounted for, heat is ground and hybridized for parent with China, passes through single seed descent, obtains the strain for stablizing heredity, forms RIL group
(obtaining 134 strains for stablizing heredity, collectively constitute RIL group)；2) using artificial infection idenfication method measurement RIL group
Disease resistance distinguishes each strain pair as quantitative trait phenotypes value, and with this using the difference in height taken over bacterium and do not connect bacterium plant
The anti-perception of Pseudomonas panici Stapp；3) a large amount of SNP markers of laboratory early development and Indel label building heredity are utilized
Map is analyzed the SNP marker of whole chromosome group and the relationship of quantitative trait phenotypes value by R-QTL, QTL is navigated to one by one
The corresponding position of linkage group, and estimate its hereditary effect.If detecting the molecular labeling of LOD > 3, then it is assumed that LOD value highest point pair
There are 1 QTL between 2 labels answered.
The present invention also provides a kind of method of artificial infection idenfication rice plant disease resistance, accounted for China, heat is ground and its offspring
134 recombinant inbred lines of group are material, artificial infection brown streak bacterium RS-1, to the plant height of each strain of parents and group into
Row measurement and quantitative analysis, it is resistant to Pseudomonas panici Stapp to judge whether plant has.
It is a further object of the present invention to provide points of the main effect QTL site Apam-1 of adjusting and controlling rice bacterial brown streak resistance
Son label and its application.
Using the primer pair of molecular labeling Indel Bbsr-1, the primer pair of molecular labeling Indel Bbsr-2, mirror is treated
The DNA for determining rice material carries out PCR amplification, and amplified production carries out electrophoresis detection, such as amplifies the DNA fragmentation of corresponding size, then
Indicate that QTL Apam-1 exists.
Wherein, the primer pair of the molecular labeling Indel Bbsr-1, after carrying out PCR amplification to DNA, electrophoresis can be detected
The band of same position is ground with rice varieties heat.
Wherein, the primer pair of the molecular labeling Indel Bbsr-1, are as follows:
Upstream primer 5 '-CATAGCATTTTCGATTTTTGAA-3 '
Downstream primer 5 '-CTTGTGACGGAACCAAATCA-3 '.
Wherein, the primer pair of the molecular labeling Indel Bbsr-2, after carrying out PCR amplification to DNA, electrophoresis can be detected
The band of same position is ground with rice varieties heat.
Wherein, the primer pair of the molecular labeling Indel Bbsr-2, after carrying out PCR amplification to DNA, electrophoresis can be detected
The band of same position is ground with rice varieties heat.
Wherein, the primer pair of the molecular labeling Indel Bbsr-2, are as follows:
Upstream primer 5 '-AATGATGAGGGGAGTGGATG-3 '
Downstream primer 5 '-AGCTACTGGTGGTGCTTGCT-3 '.
Wherein, the primer pair of the molecular labeling Indel Bbsr-2, after carrying out PCR amplification to DNA, electrophoresis can be detected
The band of same position is ground with rice varieties heat.
Wherein, the PCR reaction system is as follows
Wherein, PCR reaction condition are as follows: 94 DEG C of denaturation 3min, 94 DEG C of 30s, 57 DEG C of 30s, 72 DEG C of 30s expand 38 circulations,
Last 72 DEG C extend 10min eventually.
It is also an object of the present invention to provide the main effect QTL sites of the adjusting and controlling rice bacterial brown streak resistance
Molecular labeling breeding bacteria resistance brown streak rice varieties application.
The present invention uses the molecule labelling method of QTL site, and this method is to grind on No. 1 chromosome using what is screened in heat
2 pairs of molecular labelings of the main effect QTL Apam-1 close linkage of the adjusting and controlling rice bacterial brown streak resistance navigated to predict water
The bacterial brown streak resistance of rice material accelerates the selection progress of resistant rice varieties.
Beneficial effects of the present invention essentially consist in that:
Present invention firstly discloses the main effect QTLs of an adjusting and controlling rice bacterial brown streak resistance, influence rice plant pair
The resistance of Pseudomonas panici Stapp；The unnamed gene of adjusting and controlling rice bacterial brown streak resistance existing for the QTL site
For Apam-1.The QTL site is related to Pseudomonas panici Stapp resistance, the main growth and development for influencing plant, to plant
Height has a significant impact, to have an important influence on to rice yield and quality, it is thin which enhances plant pair rice
The resistance of bacterium property brown streak can be used for rice molecular assistant breeding, accelerate rice breeding process, simple and easy to do, safely and effectively, have
Beneficial to improving the economic value of rice varieties, it is suitable for large-scale promotion application.
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 the main effect QTL localization method of adjusting and controlling rice bacterial brown streak resistance.
Fig. 2 is the statistics histogram after artificial infection brown streak bacterium to plant height in RIL group, to judge plant pair
The Resistant Difference of Pseudomonas panici Stapp.Wherein, parent China accounts for the plant height ground with parent's heat after HZ RY representative connects bacterium
Degree, RY representative do not connect the plant height that bacterium parent's heat is ground, and HZ representative does not connect the plant height that bacterium parent China accounts for.
Fig. 3 is the primer pair of molecular labeling Indel Bbsr-1 in parent and its F1Amplification generates in generation and RIL group
Electrophoretogram.Wherein, 1 be susceptible variety China account for, 2 be disease-resistant variety heat grind, 3 be China account for/heat grind filial generation F1,4-10 be China
Account for/heat grinds the RIL group of cross combination.
Fig. 4 is the primer pair of molecular labeling Indel Bbsr-2 in parent and its F1Amplification generates in generation and RIL group
Electrophoretogram.Wherein, 1 be susceptible variety China account for, 2 be disease-resistant variety heat grind, 3 be China account for/heat grind filial generation F1,4-10 be China
Account for/heat grinds the RIL group of cross combination.
Fig. 5 is the primer pair of molecular labeling Indel Bbsr-1 parent China accounts for, heat is ground and its F1It generation and is taken turns with Hua Zhanwei
Return the electrophoretogram that generation is expanded in the BC3F1 generation of parent.Wherein, 1 is that susceptible variety China accounts for, 2 be that disease-resistant variety heat is ground, 3 be China
Account for/heat grinds filial generation F1, 4-10 be with the BC of Hua Zhanwei recurrent parent3F1The electrophoretogram of generation is expanded in generation.
Fig. 6 is the primer pair of molecular labeling Indel Bbsr-2 parent China accounts for, heat is ground and its F1It generation and is taken turns with Hua Zhanwei
Return the electrophoretogram that generation is expanded in the BC3F1 generation of parent.Wherein, 1 is that susceptible variety China accounts for, 2 be that disease-resistant variety heat is ground, 3 be China
Account for/heat grinds filial generation F1, 4-10 be with the BC of Hua Zhanwei recurrent parent3F1The electrophoretogram of generation is expanded in generation.
Fig. 7 is position of the main effect QTL Apam-1 of adjusting and controlling rice bacterial brown streak resistance on No. 1 chromosome.
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
Local rice varieties China accounts for the better resistance to Pseudomonas panici Stapp, with Hua Zhanwei donor parents, local water
Rice varieties heat is ground as receptor parent, and hybridization building RILs is carried out, using single seed descent (that is, to F1 carry out bagging single plant by kind at
Reason, until offspring's strain phenotype does not separate), (F13, all strain phenotypes are steady for the final strain for obtaining 134 stable heredity
It is fixed), such as Fig. 1.
Parent and each 60 of each strain (F13) seed 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, artificial bacteria inoculation's identification
Take RS-1 bacterial strain (4 DEG C of preservations), on NA culture medium flat plate after scribing line culture 48h, the single colonie on picking plate
Be inoculated on the NB culture medium of 200mL, with the speed of 180r/min in shaken cultivation on 28 DEG C of constant-temperature tables for 24 hours, gained bacterium solution
(concentration is 9 × 109A/ml) it can be used to connect bacterium.In rice seedling, brown streak bacterium is inoculated with using stem's injection method, when inoculation
Respectively 4 groups of experimental groups of setting and 4 groups of control groups.Starting every 2 days observation rice surface brown scabs after being inoculated with 1 week, a situation arises,
The plant height that each strain takes over bacterium and do not connect bacterium rice is measured after connecing bacterium 3 weeks, and resisting between plant is judged according to Plant Height of Rice
Sex differernce (Fig. 2).
In Fig. 2, parent China accounts for the plant height ground with parent's heat after HZ RY representative connects bacterium, and RY representative does not meet bacterium parent
The plant height that heat is ground, HZ representative do not connect the plant height that bacterium parent China accounts for；According to fig. 2 it can be seen that after connecing bacterium 3 weeks, the strain of parent
High variation difference is obvious.It is 104.00cm that HZ, which connects the average plant height after bacterium, and the average plant height than not connecing bacterium plant normally is short
6.50cm；The average plant height of RY is 101.40cm, shorter than the average plant height of normal plant 3.60cm, brown streak between two parents
Resistant Difference is obvious.Plant height statistical result showed, it is 105.40cm that RIL group, which connects the average plant height after bacterium, normal plant it is flat
Equal plant height is 110.43cm.Occur a certain number of super close individuals in group, and plant height shows as normal distribution.
RS-1 bacterial strain is having disclosed the IcmF and DotU are required for being published in " Arch.Microbiol "
There is informing in for the virulence of Acidovorax oryzae strain RS-1.
Embodiment 3, QTL positioning analysis
Using the genetic map of a large amount of SNP markers and Indel the label building of laboratory early development, bacterium is connect to brown streak
Plant height afterwards carries out quantitative trait loci (QTL) Interval mapping, and the SNP of whole chromosome group is analyzed by professional software R-QTL
QTL, is navigated to the corresponding position of linkage group, and estimate its hereditary effect by the relationship of label and quantitative trait phenotypes value one by one.
If detecting the molecular labeling of LOD > 3, then it is assumed that there are 1 QTL (Fig. 3,4) between corresponding 2 labels of LOD value highest point.?
A main effect QTL on No. 1 chromosome, genetic distance 162.95-172.83cM, object are found in whole chromosome group
Reason distance is 38013068bp-40317380bp, and is named as Apam-1 (Fig. 7).
QTL refers to controlling the size position of the gene in genome of character, and the QTL that the present invention positions is located at No. 1 dye
On colour solid DNA fragmentation, physical distance is between 38013068bp-40317380bp.
Embodiment 4, accounted for China, heat is ground and its total 2680 plants of progeny population (20 repetitions are arranged in every class), artificial infection
Brown streak bacterium RS-1, accounts for parents China, heat is ground and the plant height of each strain of group measures and quantitative analysis, judges plant
Whether have resistant to Pseudomonas panici Stapp；
It is specific as follows: to be carried out after connecing bacterium 3 weeks, height of the rice from stalk bottom to highest point, each strain is measured with meter scale
System measures the rice that 4 plants of rice for taking over bacterium and 4 plants do not take over bacterium respectively, seeks average respectively, subtracts each other to obtain the strain and connects
The difference in height of plant before and after bacterium.
Judged according to following criterion: when the rice for taking over bacterium in the strain is averaged plant height/do not connect bacterium water
Rice be averaged plant height > 0.96 when, determine that the strain is resistant to Pseudomonas panici Stapp；Conversely, when the strain is inscribed
Cross bacterium rice be averaged plant height/do not connect bacterium rice be averaged plant height≤0.96 when, then determine the strain to rice bacterium
Property brown streak do not have resistance.
Resulting measurement result such as Fig. 2.
Confirmatory experiment, the verifying for determine to above-mentioned plant correctness using molecular marker assisted selection method:
Molecular labeling Bbsr-1 and molecular labeling Bbsr-2 is set separately in QTL site Apam-1 upstream and downstream.
The primer pair of molecular labeling Indel Bbsr-1 are as follows:
Upstream primer 5 '-CATAGCATTTTCGATTTTTGAA-3 '
Downstream primer 5 '-CTTGTGACGGAACCAAATCA-3 '；
The primer pair of molecular labeling Indel Bbsr-2 are as follows:
Upstream primer 5 '-AATGATGAGGGGAGTGGATG-3 '
Downstream primer 5 '-AGCTACTGGTGGTGCTTGCT-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, PCR reaction system: 1 μ L of upstream primer, 1 μ L, DNA2 μ L, mix enzyme of downstream primer to its genomic DNA
6μL.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, amplification 38
A circulation, last 72 DEG C extend 10min eventually.Pcr amplification product is detected with 4% agarose gel electrophoresis.
As a result as follows: it is resistant to be determined pairs of Pseudomonas panici Stapp, verifies resulting result through the method and is
The electrophoretic band of pcr amplification product is intended to parent's heat and grinds；It is determined into not resistant to Pseudomonas panici Stapp, warp
The method verifies the electrophoretic band that resulting result is pcr amplification product and grinds (Fig. 5,6) different from parent's heat.That is, two methods institute
The result obtained is completely the same.
The application of embodiment 5, Rice Resistance brown streak 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 rice varieties of sense brown streak grind with heat and are hybridized if China accounts for, and obtain corresponding F1, returned with Hua Zhanwei recurrent parent
It hands over, until BC3F1Generation.Extract BC3F1For part single plant DNA, PCR then is carried out with the primer of Indel label Bbsr-1 and Bbsr-2
Amplification, corresponding label is determined whether there is by band analysis, there are the explanation of the label brown items of strain paddy bacterial
Disease reaches anti-level, and there is no be then susceptible (Fig. 4).The selfed seed of positive single plant is collected after label auxiliary detection, then benefit
Manually the method for inoculated identification carries out the brown streak Resistance Identification in seedling stage.Verification result is that the plant of conversion has rice bacterium
Property brown streak resistance, connect plant height/normal plant > 0.96 after bacterium, that is, obtain the water with brown streak resistant gene Apam-1
Rice transgenic line.Screened using this method, M8003 line, can get with brown streak resistance and remain China be dominant it is benign
The rice of shape, substantially increases breeding efficiency.
In conclusion the main effect QTL of adjusting and controlling rice bacterial brown streak resistance of the invention can effectively enhance rice
The resistance of plant pair Pseudomonas panici Stapp can decrease or even eliminate Pseudomonas panici Stapp to water in breeding process
The harm of rice plants, while can accelerate to optimize the process of rice varieties.It simultaneously can be with during rice molecular assistant breeding
Anti- brown streak rice is cultivated, the disease incidence of Pseudomonas panici Stapp is reduced.Such method is simple, safely and effectively, beneficial
In the economic value for improving rice varieties, it 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
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Claims (5)
1. the main effect QTL Apam-1 of adjusting and controlling rice bacterial brown streak resistance, it is characterised in that: QTL site is located at No. 1 dyeing
On body DNA fragmentation, genetic distance 162.95-172.83cM, physical distance 38013068bp-40317380bp.
2. the Indel of the main effect QTL site Apam-1 of adjusting and controlling rice bacterial brown streak resistance is marked, it is characterised in that: be
The primer pair of Bbsr-1 and Bbsr-2；
The primer pair of molecular labeling Indel Bbsr-1 are as follows:
Upstream primer 5 '-CATAGCATTTTCGATTTTTGAA-3 '
Downstream primer 5 '-CTTGTGACGGAACCAAATCA-3 '；
The primer pair of molecular labeling Indel Bbsr-2 are as follows:
Upstream primer 5 '-AATGATGAGGGGAGTGGATG-3 '
Downstream primer 5 '-AGCTACTGGTGGTGCTTGCT-3 '.
3. a kind of method of artificial infection idenfication rice plant disease resistance, it is characterised in that:
Accounted for using China, heat is ground and its progeny population is research object, artificial infection brown streak bacterium RS-1, parents China is accounted for, heat is ground and
The plant height of each strain of group measures and quantitative analysis, judges whether plant has and have to Pseudomonas panici Stapp
Resistance.
4. the method for artificial infection idenfication rice plant disease resistance according to claim 3, it is characterised in that criterion
Are as follows: when the rice that bacterium is taken in the strain be averaged plant height/do not connect bacterium rice be averaged plant height > 0.96 when, determine the strain
It is resistant to Pseudomonas panici Stapp；The plant height/do not connect bacterium the water conversely, rice for taking over bacterium in the strain is averaged
Rice be averaged plant height≤0.96 when, then determine that the strain does not have resistance to Pseudomonas panici Stapp.
5. the molecular labeling application of the main effect QTL of an adjusting and controlling rice bacterial brown streak resistance, which is characterized in that utilize molecule
Labeling method is screened, M8003 line, be can get the rice with cecospora spot resistance, is substantially increased breeding efficiency.
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| 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 |
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