CN110178719A - A method of improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance - Google Patents
A method of improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance Download PDFInfo
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- CN110178719A CN110178719A CN201910409982.2A CN201910409982A CN110178719A CN 110178719 A CN110178719 A CN 110178719A CN 201910409982 A CN201910409982 A CN 201910409982A CN 110178719 A CN110178719 A CN 110178719A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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Abstract
The invention discloses a kind of methods for improveing Rice Resistance To Rice Blast and bacterial leaf spot resistance, hybridize Bacterial blight resistance gene and blast resistant gene is carried simultaneously with the rice material for not carrying above-mentioned resistant gene, seedling stage foreground detection is carried out using SNP marker, selection carries the single plant of Bacterial blight resistance gene and blast resistant gene, and be selfed after using rice material of the resistance wait improveing carrying out continuous backcross as recurrent parent, obtain bacterial leaf-blight and rice blast resistance improvement and comprehensive agronomy character improved materials similar with recurrent parent.Pass through the high filial generation of the SNP marker screening blast resisting, Bacterial blight resistance gene and background response rate of close linkage, realize polymerization rice blast, bacterial leaf spot resistance gene not only to achieve the purpose that improve seed rice disease resistance, and improved materials are similar to recurrent parent, breeding cycle is short, saves cost.
Description
Technical field
The present invention relates to rice breeding technology fields, in particular to a kind of to improve Rice Resistance To Rice Blast and bacterial leaf spot simultaneously
The method of sick resistance.
Background technique
Rice is the important cereal crops in China.Rice blast and bacterial leaf-blight are the Major Diseases of hazard rice, and harm is tight
It can lead to the rice underproduction 30-50% when weight even to have no harvest.Hybrid rice superior restorer line R900 is that super hybridized rice tackling key problem generates
Great innovation achievements, matched cross combination " super excellent No. thousand " mostly are creating high yield record in recent years.But restorer R900
And its match the high sense bacterial leaf-blight of combination and high sense rice blast, exist during promoting and applying because bacterial leaf-blight or rice blast are broken out
And the risk of Severe Reduction.
Cultivate resistant rice varieties be always breeding circle generally acknowledge prevention and treatment rice blast and bacterial leaf-blight it is most economical
Effective approach.The present invention proposes a kind of based on full-length genome SNP marker, the white leaf of slewing improvement restorer R900
The molecular breeding method of blight and rice blast resistance.Using wide spectrum bacterial blight-resisting and blast resisting major gene resistance, pass through combination
With the SNP marker foreground selection and full-length genome SNP marker Foreground selection of resistance target gene close linkage, quickly
The bacterial leaf spot resistance and rice blast resistance of orderly improvement restorer R900, while characteristics, the realization such as its high yield, high-quality being kept to change
Good variety resistance and the coordinating and unifying for keeping original good characteristic.The invention achievement will for hybrid rice restorer line R900 and its
Cross combination further safely, fully play advantage and large scale application provides safeguard.
Summary of the invention
The present invention is using the utilization SNP molecule mark chain with bacterial leaf spot resistance gene and rice blast resistance gene
Note, selection hybridize above-mentioned resistant gene present in segregating population with bacterial blight-resisting and rice blast material to improved materials, and
Full-length genome SNP marker Foreground selection is carried out to the plant of resistant gene, it is white with breeding in conjunction with conventional breeding means
Leaf blight and rice blast resistance significantly improve, the breeding side of comprehensive agronomy character and the rice material almost the same to improved materials
Method.
In order to achieve the above objectives, the technical scheme is that passing through following step realization:
A method of improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance, comprising the following steps:
(1) recurrent parent hybridizes with the rice material for carrying bacterial leaf spot resistance gene and rice blast resistance gene, obtains hybrid
F1For seed；
(2) hybrid F is planted1For seed, pseudostationary plant is excluded using SNP marker, by true hybrid F1Generation and recurrent parent
Backcrossing obtains BC1F1For seed；
(3) BC is planted1F1For seed, divide using with the SNP of bacterial leaf spot resistance gene and rice blast resistance gene close linkage
Son label is to BC1F1Seedling stage foreground detection is carried out for group plant, selects while carrying bacterial leaf spot resistance and rice blast resistance base
The BC of cause1F1For single plant, genetic background inspection is carried out to the plant containing resistant gene followed by full-length genome SNP marker
It surveys, selects the highest single plant of background response rate and recurrent parent continuous backcross, each backcross generations select to carry bacterial leaf-blight
Resistance and rice blast resistance gene and the highest single plant of background response rate are returned with recurrent parent, backcrossing algebra be m(m≤
2) backcross progeny BC, is obtainedmFl；
(4) BC is plantedmFlIt for seed, is detected using SNP marker, selection carries bacterial leaf spot resistance gene and rice blast is anti-
Property gene and the highest BC of background response ratemFlFor individual plant selfing, selfing algebra is n(n≤2), it obtains comprehensive agronomy character and stablizes
BCmFnFor seed；
(5) BC is plantedmFnIt for seed, is detected with SNP marker, select while carrying bacterial leaf spot resistance and rice blast resistance
The BC of homozygous genemFnFor single plant, obtains bacterial leaf-blight and rice blast resistance improvement and comprehensive agronomy character is similar to recurrent parent
Improved materials.
Further, the recurrent parent is hybrid rice restorer line R900.
Further, the bacterial leaf spot resistance gene includes at least one of Xa23, Xa21, Xa7.
Further, the rice blast resistance gene includes at least one of Pi1, Pi2, Pi9, Pigm.
Further, the step (3), step (4), take in the single plant that is selected by SNP marker in step (5)
Band bacterial leaf spot resistance gene has 1-3, while carrying rice blast resistance gene has 1-2 a.
Further, in the step (1), while the rice of bacterial leaf spot resistance gene and rice blast resistance gene is carried
Any one breeding material of material in long-grained nonglutinous rice, japonica rice, restorer, holding system.
Further, the step (2), step (3), step (4), step (5) detect bacterial leaf spot using SNP marker
The purpose that the single plant of sick resistant gene and rice blast resistance gene is returned or is selfed is, it is ensured that above-mentioned resistant gene is being improved
Exist always in offspring.
Further, the step (3), in step (4), SNP full-length genome Markers for Detection background is utilized to reply
Rate, obtains the highest plant of background response rate and the purpose of recurrent parent continuous backcross is, faster obtains comprehensive agronomy
Shape improved materials similar with recurrent parent.
Further, the step (2), step (3), step (4), in step (5), detection bacterial leaf spot resistance gene and
The SNP marker self-developing of rice blast resistance gene.
Further, the step (3), in step (4), detect the full-length genome SNP marker of background response rate from
Row exploitation.
Beneficial effects of the present invention:
(1) breeding material that the present invention finally obtains has polymerize rice blast resistance gene and bacterial leaf spot resistance gene；
(2) present invention is marked when detecting resistant gene using the SNP marker of close linkage, while using full-length genome SNP
It remembers row Foreground selection into and comprehensive agronomy character is selected, can significantly shorten breeding cycle, improve breeding efficiency；
(3) it is significantly increased on the rice blast resistance and bacterial leaf spot resistance of the invention for making the breeding material finally obtained, and with
Economical character to improved materials is consistent substantially.
Specific embodiment
It should be noted that in the absence of conflict, the feature in embodiment and embodiment in the present invention can phase
Mutually combination.
It is described in detail with a specific embodiment: utilizing bacterial leaf spot resistance gene entrained by HZ02455
The bacterial leaf-blight and rice blast resistance of Xa23 and rice blast resistance gene Pi9 orderly improvement hybrid rice restorer line R900, this hair
Bright people does not carry Xa23 and Pi9 resistant gene using SNP marker detection display restorer R900, and HZ02455 is carried
Xa23 and Pi9 resistant gene, specific implementation step are as follows:
1, restorer R900 is hybridized with HZ024550, obtains 56, F-1 hybrids seed.When hybridization, mother is with restorer R900
Originally, HZ024550 does male parent or HZ024550 is maternal, restorer R900 and does male parent；
It does male parent preferably, the present embodiment is maternal, HZ024550 with restorer R900 and is hybridized to obtain F1 generation.
2, above-mentioned F1 generation seed is planted, whether carries bacterial blight-resisting using molecular markers for identification F1 generation single plant in Seedling Stage
Gene Xa23 and blast resistant gene Pi9 obtains true 24 plants of hybrid of F1 generation to exclude pseudostationary；Select the true hybrid of F1 for male parent with
Restorer R900 backcrossing, obtains BC1F1772, seed.
Plant BC1F1For seed, Seedling Stage identified using SNP marker and meanwhile Bacterial blight resistance gene Xa23 and
The heterozygosis single plant of blast resistant gene Pi9 amounts to 562 plants；Later, full-length genome SNP marker is utilized to the single plant selected
Genetic background selection is carried out to target gene heterozygosis single plant, selects background response rate highest, and at heading stage and restorer
The single plant that the economical character of R900 is most like makees male parent, is returned with restorer R900, obtains 1097, BC2F1 seed.
Genetic background is detected, if recipient genotypes are AA, is denoted as " 0 ", and donor gene type is BB, is denoted as " 2 ", miscellaneous
Conjunction genotype is AB, is denoted as " 1 ", background response rate=(a*2+b)/[(a+c+b) * 2].
Wherein a represents the sum of " 0 ", and b represents the sum of " 1 ", and c represents the sum of " 2 "；
BC2F1 seed is planted, is identified in Seedling Stage using SNP marker and is contained Bacterial blight resistance gene Xa23 simultaneously and resist
The heterozygosis single plant of rice blast ospc gene Pi9 amounts to 747 plants；Later, full-length genome SNP marker pair is utilized to the single plant selected
Target gene heterozygosis single plant carries out genetic background selection, selects background response rate highest, i.e., in maturity period and restorer R900
Most like 3 single plants of economical character, sowing respectively after individual plant selfing obtains the BC of 3 strains2F22984, seed.
5, BC2F2 seed is planted, identifies Bacterial blight resistance gene Xa23 and anti-rice using SNP marker in Seedling Stage
Seasonal febrile diseases gene Pi9 homozygous single plant, amounts to 116 plants；Later, full-length genome SNP marker is recycled to the single plant selected
Genetic background detection is carried out to target gene homozygosis single plant, selects background response rate highest, and in maturity period and restorer R900
Most like single plant 9 of economical character, sowing is distinguished after individual plant selfing, obtains 5884, BC2F3 seed of 9 strains.
6, each strain selects 20 seeds respectively, plants the BC2F3 seed of 9 strains, utilizes SNP molecule in Seedling Stage
Marker Identification goes out the homozygous single plant of Bacterial blight resistance gene Xa23 and blast resisting target gene Pi9, selects to obtain group whole
Homogeneous causes, Comprehensive Traits and the similar strain of restorer R900 3, every strain select 1 typical single plant, are selfed independent sowing, obtain
Obtain 3016, BC2F4 seed of 3 strains.
7, the BC2F4 seed of 3 strains is planted, while carrying out bacterial leaf-blight inoculation and rice blast sick nursery resistance mirror in Hunan
It is fixed, identify high resistance to hoja blanca and rice blast, group's stable and consistent, Comprehensive Traits and the most like strain 1 of restorer R900
It is a.
Specifically, blast resistance identification: endanger serious rice region throughout the year in rice blast, select soil fertility level it is high,
The convenient rice field of irrigation and drainage is as sick nursery, convenient for management and disease survey.For trying identification of species sowing quantity 5-10g, capable length is each broadcast
50cm, wide 10cm, 10 cm of line space are randomly provided three repetitions, induce susceptible variety mixed seeding for examination identification of species
Around.In seedling length to two one heart stage of leaf, the blade that rice blast infects is collected in other fields of sick nursery, 2-3cm is cut into, through protecting
After warm 24 h of moisturizing, it is spread on and induces in kind.30 ~ 35d length of time rice seedlings grow is transplanted, every 2 row of variety transplanting, every row 7 clumps, and every kind is total
14 clumps；Seeding row spacing is 20cm；Ginseng inserts each 1 part of anti-sense kind between every 10 kinds.Kind of a protection row, strain are inserted in ginseng mirror kind surrounding
Line-spacing is identical as ginseng mirror kind；Row kind is protected to use susceptible variety.Seedling pest 4 ~ 6 leaf phases before rice transplanting are investigated, leaf
Pest tillering regularity after rice transplanting is investigated.Each kind falls ill most heavy 10 plants as respondent, every plant of investigation hair
The most heavy blade of disease takes foundation of 3 plants most heavy of the average value as variety evaluation of falling ill.Susceptible control miaoye seasonal febrile diseases grade is not
Reach 4 grades, group test is invalid.Fringe pest is investigated to dough stage after Rice Heading, each identification of species cell, investigation morbidity is most
The spike of rice of weight, no less than 100 fringes count fringe pest disease incidence, single fringe fringe pest loss late.Susceptible control tassel blast grade is not up to 7 grades
, group test is invalid.
Bacterial leaf spot resistance identification: 20 days before Rice Heading, the generation of rice leaf spot bacteria advantage pathological form bacterium is selected
Table bacterial strain.Bacterial strain is cultivated 72 hours under the side of body 28 DEG C of constant temperature of Ben Zheshi potato semisynthetic medium, with bacterium under sterile washing
Bacterium bacteria suspension is diluted to 10 with wheat formula turbidimetry by tongue fur8~109Cell/milliliter bacterium solution.Preferably in fine day or cloudy afternoon, will connect
Kind body cuts off 2 cm or so at the top of the full lamina in rice strain top with leaf-cutting method.It investigates within 20~25 days or so after inoculation, wherein susceptible product
The sick grade of kind, which reaches 7 grades (sense) or more, can think when time identification is effective.
Screening: the strain of high resistance to hoja blanca and rice blast is identified according to the incidence of rice material.
The strain is to have imported two resistant genes of Pi9 and Xa23 and economical character and and restorer R900 phase simultaneously
As finally improve strain.
Likewise, using the bacterial leaf spot resistance gene Xa21 or Xa7 and rice blast resistance gene Pi 1 or Pi2 carried or
Other breeding materials of Pigm can also obtain improveing strain accordingly according to the above method.
The SNP marker that detection Bacterial blight resistance gene Xa23 and blast resisting target gene Pi9 whether there is is opened
Forwarding method is the DNA sequence dna self-developing according to reported Xa23 and Pi9 resistant gene.
Detect the method whether a rice single plant carries two resistant genes of Xa23 and Pi9 are as follows: utilize independently developed 4
A specificity SNP marker detects the leaf DNA of the single plant respectively, if the SNP marker of 4 close linkages can detect simultaneously
For advantageous (homozygosis) genotype, then the single plant carries two resistant genes of Xa23 and Pi9 simultaneously.
The experimental techniques such as SNP detection of the present invention are competitiveness allele PCR(Kompetitive Allele
Specific PCR, KASP) principle, reaction detection carries out in Array Tape genotyping platform, and this method is answered successively
For work such as molecular mark, the objective trait assignment of genes gene mapping, seed purity and authenticity identifications, have it is at low cost,
The advantages such as flux height and fluorescence signal acquisition data are accurate.Matched reagent consumptive material is purchased from LGC company, Britain.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method for improveing Rice Resistance To Rice Blast and bacterial leaf spot resistance, which comprises the following steps:
(1) recurrent parent hybridizes with the rice material for carrying bacterial leaf spot resistance gene and rice blast resistance gene, obtains hybrid F1
For seed；
(2) hybrid F is planted1For seed, pseudostationary plant is excluded using SNP marker, by true hybrid F1In generation, returns with recurrent parent
It hands over, obtains BC1F1For seed；
(3) BC is planted1F1For seed, the SNP molecule mark of bacterial leaf spot resistance gene and rice blast resistance gene close linkage is utilized
Note is to BC1F1Seedling stage foreground detection is carried out for group plant, selection carries bacterial leaf spot resistance and rice blast resistance gene
BC1F1For single plant, genetic background detection, choosing are carried out to the plant containing resistant gene followed by full-length genome SNP marker
Select the highest single plant of background response rate and recurrent parent continuous backcross, each backcross generations select to carry bacterial leaf spot resistance and
The rice blast resistance gene and highest single plant of background response rate is returned with recurrent parent, backcrossing algebra are m(m≤2), it obtains
Backcross progeny BCmFl；
(4) plantation BCmFl is detected for seed using SNP marker, and selection carries bacterial leaf spot resistance gene and rice blast is anti-
Property gene and the highest BCmFl of background response rate be selfed for single plant, selfing algebra be n(n≤2), obtain comprehensive agronomy
The stable BCmFn of shape is for seed；
(5) plantation BCmFn is detected for seed with SNP marker, and selection carries bacterial leaf spot resistance and rice blast resistance is homozygous
The BCmFn of gene obtains bacterial leaf-blight and rice blast resistance improvement and comprehensive agronomy character is similar with recurrent parent for single plant
Improved materials.
2. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
Stating recurrent parent is hybrid rice restorer line R900.
3. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
Stating bacterial leaf spot resistance gene includes at least one of Xa23, Xa21, Xa7.
4. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
Stating rice blast resistance gene includes at least one of Pi1, Pi2, Pi9, Pigm.
5. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
Step (3), step (4) are stated, bacterial leaf spot resistance gene is carried in the single plant that is selected in step (5) by SNP marker and has
1-3, and carrying rice blast resistance gene has 1-2 a.
6. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
State in step (1), at the same carry the rice breed of bacterial leaf spot resistance gene and rice blast resistance gene from long-grained nonglutinous rice,
Japonica rice, restorer, keep system in any one.
7. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
It states step (2), step (3), step (4), step (5) and utilizes SNP marker detection bacterial leaf spot resistance gene and rice blast
The purpose that the single plant of resistant gene is returned or is selfed is, it is ensured that above-mentioned resistant gene exists always in improved progeny.
8. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
It states step (3), in step (4), utilizes SNP full-length genome Markers for Detection background response rate, obtain background response rate highest
Plant and the purpose of recurrent parent continuous backcross be faster to obtain the improvement similar with recurrent parent of comprehensive agronomy character
Material.
9. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that institute
It states step (2), step (3), step (4), in step (5), detect the SNP of bacterial leaf spot resistance gene and rice blast resistance gene
Molecular labeling self-developing.
10. the method for improvement Rice Resistance To Rice Blast and bacterial leaf spot resistance according to claim 1, which is characterized in that
In the step (3), step (4), the full-length genome SNP marker self-developing of background response rate is detected.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112806252A (en) * | 2019-11-15 | 2021-05-18 | 华智水稻生物技术有限公司 | Method for creating multi-resistance rice new germplasm by using molecular marker |
| CN115735756A (en) * | 2022-10-18 | 2023-03-07 | 长江大学 | Breeding method of disease-resistant rice variety |
| CN116210585A (en) * | 2022-08-26 | 2023-06-06 | 海南省农业科学院粮食作物研究所 | Method for molecular marker assisted selection of Hainan disease and pest resistant rice restorer line |
| CN116789786A (en) * | 2022-09-14 | 2023-09-22 | 云南省农业科学院生物技术与种质资源研究所 | Rice Xa48 (t) protein and application of coding gene thereof |
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| CN103931490A (en) * | 2014-04-04 | 2014-07-23 | 福建农林大学 | Breeding method of resistance-variable rice variety |
| CN104770287A (en) * | 2015-04-10 | 2015-07-15 | 中国水稻研究所 | Method for improving paddy blast resistance by using mid-early 39 |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103931490A (en) * | 2014-04-04 | 2014-07-23 | 福建农林大学 | Breeding method of resistance-variable rice variety |
| CN104770287A (en) * | 2015-04-10 | 2015-07-15 | 中国水稻研究所 | Method for improving paddy blast resistance by using mid-early 39 |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112806252A (en) * | 2019-11-15 | 2021-05-18 | 华智水稻生物技术有限公司 | Method for creating multi-resistance rice new germplasm by using molecular marker |
| CN112806252B (en) * | 2019-11-15 | 2022-08-02 | 华智生物技术有限公司 | Method for creating multi-resistance rice new germplasm by using molecular marker |
| CN116210585A (en) * | 2022-08-26 | 2023-06-06 | 海南省农业科学院粮食作物研究所 | Method for molecular marker assisted selection of Hainan disease and pest resistant rice restorer line |
| CN116789786A (en) * | 2022-09-14 | 2023-09-22 | 云南省农业科学院生物技术与种质资源研究所 | Rice Xa48 (t) protein and application of coding gene thereof |
| CN116789786B (en) * | 2022-09-14 | 2023-11-10 | 云南省农业科学院生物技术与种质资源研究所 | Rice Xa48 (t) protein and application of coding gene thereof |
| CN115735756A (en) * | 2022-10-18 | 2023-03-07 | 长江大学 | Breeding method of disease-resistant rice variety |
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