CN106106156A - A kind of breeding method of anti-rough dwarf disease Semen Maydis - Google Patents
A kind of breeding method of anti-rough dwarf disease Semen Maydis Download PDFInfo
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- CN106106156A CN106106156A CN201610493515.9A CN201610493515A CN106106156A CN 106106156 A CN106106156 A CN 106106156A CN 201610493515 A CN201610493515 A CN 201610493515A CN 106106156 A CN106106156 A CN 106106156A
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Abstract
The present invention discloses the breeding method of a kind of anti-rough dwarf disease Semen Maydis, by the method for hybridization, anti-rough dwarf disease gene is introduced corn breeding trunk strain material, cultivate a kind of anti-rough dwarf disease corn strain, prior art is avoided to utilize the environmental pollution that the anti-rough dwarf disease of Agro-chemicals control Semen Maydis causes, shorten breeding cycle, increase Semen Maydis anti-rough dwarf disease ability while keeping breeding excellent properties, reduce Maize Production cost, there is application prospect.
Description
Technical field
The present invention relates to a kind of breeding method, particularly relate to a kind of breeding method based on anti-rough dwarf disease Semen Maydis, belong to existing
For agricultural high-tech field.
Background technology
Semen Maydis is long at the cultivation history of China, is important grain and forage crop, has yield height, shelf-stable, again
The feature that productive value is high, but along with climate warming, cropping system change and the multiple adverse circumstance such as single variety establishing in large scale because of
Element causes or increases the weight of the pest and disease damage of corn crop, has had a strong impact on the yield stability of Semen Maydis.
MRDV is the virosis of serious harm Semen Maydis stable yields in China.Present system after Semen Maydis is susceptible to invade
Dye, growth promoter is obstructed, the Maize rough dwarf virus mainly propagated with persistent fashion by small brown rice planthopper or Semen Maydis black streak dwarf
Poison causes.Nontoxic small brown rice planthopper inhales juice on the plant infected RBSDV and obtains poison, obtain poison the time be generally 24-48h, the shortest lh, nothing
Can pass poison, not transovarial passage after poisonous insect one band poison all the life, poisonous insect majority is that short-term interval passes poison.
The traditional method of preventing and treating MRDV is turned off its route of transmission, utilizes insecticide that small brown rice planthopper is carried out preventing and treating and is
Prevent and treat the effective ways of this disease.But produce upper widely used pesticide at present just not have once catch an illness just for small brown rice planthopper, plant
Having highly effective prophylactico-therapeutic measures, its effect is limited；Along with these utilization of pesticides time limits increase, insect insecticide produces
High drug-resistance；And utilization of pesticides is inevitably to environment and destruction.In production process also by
Improve the measure that MRDV is prevented and treated by cultivation technique condition, but produce little effect.
Prior art adds Maize Production to a certain extent to the Preventing Countermeasures of MRDV in Maize Production
Cost, and increased the weight of ambient pressure.
It is to select the corn variety of disease resistance that prior art prevents and treats the maximally efficient method of MRDV, cultivates Corn Crude
The method of the sick resistant variety of contracting generally has two kinds: conventional breeding and molecular engineering breeding.
The cycle of conventional breeding is the longest, cultivates the time that a MRDV resistant variety generally requires 6～8 years,
And owing to complexity and the pathogenic multiformity of the heredity of rough dwarf disease virus biological strain make the virulence gene of pathogenic bacteria colony
Composition change, often the resistant variety of selection-breeding can infect the dominant races of this kind promoting i.e. to produce after 3～5 years,
The resistance ultimately resulting in new varieties is lost.Conventional breeding elapsed time is long, especially as the increase of rough dwarf disease virus mutation frequency,
The speed that new varieties are bred as has not caught up with the mutating speed of rough dwarf disease virus.
Transgenic breeding and molecular marker assisted selection equimolecular technology Breeding are the new direction of current agricultural development, to spy
The transgenic breeding that fixed excellent main breed carries out rough dwarf disease disease-resistant gene and other excellent genes is a kind of alternative, but by
Resisted in current transgenic breeding, and transgenic breeding and molecular marker assisted selection breeding are firstly the need of gene mapping, with
Sample needs the hybridization of land for growing field crops many generations to backcross, and the cycle is the longest, and molecular engineering breeding there is also costly, technical threshold is high asks
Topic, it is difficult to grasped by general breeder.
Summary of the invention
Defect based on prior art, the present invention provides the breeding method of a kind of anti-rough dwarf disease Semen Maydis, it is possible to by rough dwarf disease
Resistant gene efficiently and exclusively penetrates in local high-grade maize kind.
Needed for the breeding method of the anti-rough dwarf disease Semen Maydis that the present invention relates to, seed material includes, the first rough dwarf disease resistant corn
Material, the second rough dwarf disease resistant corn material and corn breeding trunk strain material.
Described corn breeding trunk strain material is yield height, meter Zhi Hao, the germline of susceptible rough dwarf disease, and described first slightly contracts
Sick resistant corn material and the second rough dwarf disease resistant corn material are high anti-rough dwarf disease, not high yield, are at source not suitable on a large scale
The germline promoted.
Corn breeding trunk strain material is preferably Zheng Dan 958, and described first rough dwarf disease resistant corn material is preferably stepped on
Sea 3622, described second rough dwarf disease resistant corn material is preferably gold sea 5.
Specifically include following steps:
(1) first sowing and the program of transplanting:
First rough dwarf disease resistant corn material and the second rough dwarf disease resistant corn material are soaked seed respectively in the seed soaking of dilution
In liquid, after soaking seed 1～2 day, accelerating germination is also seeded in experiment rice seedling bed by kind, and individual plant is transplanted, and seeding row spacing is 30 × 30cm；The biggest
Field rich water and disease pest and weed management, to normal plants to blooming stage, carry out step (2).
Described seed-soaking liquid is 2mL10% dithiocyano-methane cream, 2g prochloraz, 15mL40% good fortune gram suspended seed coating
Agent, 2mL70% imidacloprid water dispersible granule, add 500 times of water dilutions formulated.
(2) acquisition of Hybrid F1:
The corncob of maternal plant is immersed in the warm water of 39～46 DEG C, take out after soaking 6～10 minutes, natural cooling 30
Minute, with paper bag the corncob of maternal plant packaged and mark, stand-by.When paternal plant enters florescence, by male parent
On plant, the flower pesticide of corncob is uniformly fallen in paper bag on the stigma of maternal plant corncob, and paper bag is continued after completing by artificial pollination
Continue and package, wait the Grain Ripening of hybrid maize fringe, i.e. obtain Hybrid F1.
The warm water temperature that the corncob of described maternal plant immerses is preferably 42 DEG C, and described soak time is preferably 7 minutes.
This method setting percentage is high, and can Selfing avoidance, moreover it is possible to Accelerate bloom, it is simple to pollination.
(3) the small brown rice planthopper screening of the anti-rough dwarf disease of Hybrid F1:
Hybrid F1 positive season is sowed, when Hybrid F1 was in for 2～2.5 leaf phase, by every strain Semen Maydis 10～the quantity of 20
Discharge small brown rice planthopper to rice seedling bed, after 50 days, every strain Semen Maydis is estimated, retain the plant not encroached on by rough dwarf disease.
Described small brown rice planthopper every feeds Maize rough dwarf virus Extracted toxin liquid 4～5 days, all carries MRDV
Poison.
(4) the antherderived callus induction of Hybrid F1:
What in the middle part of corncob, flower pesticide position accounted for grain husk shell 1/2 takes fringe for standard, takes after fringe with 70% ethanol surface sterilization, so
Encase whole corncob with plastic sheeting afterwards, keep corncob to moisten and put in refrigerator, Cold pretreatment 3 days at 4 DEG C；?
The ethanol of 85% is disinfected 2～10 minutes, with cutting off flower pesticide top after drying, flower pesticide is placed in the first culture medium, enters
Row inducing culture, light culture 2～after 3 days at 22～28 DEG C, it is transferred in the second culture medium, in 25～29 DEG C, photoperiod
Successive transfer culture is carried out under 16h7500lx illumination 8h dark, by the anther callus of the Hybrid F1 of acquisition after 50 days, statistics flower
Medicine callus induction rate, if induction of anther callus rate is less than 5%, then needs Hybrid F1 and rough dwarf disease resistant corn material
Material parent backcrosses until induction of anther callus rate is higher than 5%.
(5) second sowings and the program of transplanting:
Corn breeding trunk strain material and Hybrid F1 are soaked seed respectively in the seed-soaking liquid of dilution, after soaking seed 1-2 days,
Accelerating germination is also seeded in experiment rice seedling bed by kind, and individual plant is transplanted, and seeding row spacing is 30 × 30cm；General land for growing field crops rich water and disease pest and weed pipe
Reason, to normal plants to blooming stage, carries out step (6).
The same step of described seed-soaking liquid (1).
(6) three acquisitions handing over F1 cenospecies:
The corncob of maternal plant is immersed in the warm water of 39～46 DEG C, take out after soaking 6～10 minutes, natural cooling 30
Minute, cut off l/3 grain husk shell and do not open the little Hua of grain husk, with paper bag the corncob of maternal plant packaged and mark, stand-by.Work as father
When this plant enters florescence, the flower pesticide of corncob on paternal plant is uniformly fallen the stigma of maternal plant corncob in paper bag
On, paper bag is continued to package after completing by artificial pollination, waits the Grain Ripening of hybrid maize fringe, i.e. obtains three friendship F1 cenospecies.
The warm water temperature that the corncob of described maternal plant immerses is preferably 42 DEG C, and described soak time is preferably 7 minutes.
(7) the three small brown rice planthopper screenings handing over the anti-rough dwarf disease of F1 cenospecies:
Three hand over the sowing of F1 cenospecies positive season, when three hand over F1 cenospecies to be in for 2～2.5 leaf phase, by every strain Semen Maydis 10～20
The quantity of head discharges small brown rice planthopper to rice seedling bed, after 50 days is estimated every strain Semen Maydis, retains the plant not encroached on by rough dwarf disease.
Described small brown rice planthopper every feeds Maize rough dwarf virus Extracted toxin liquid 4～5 days, all carries MRDV
Poison.
(8) the three antherderived callus inductions handing over F1 cenospecies:
What in the middle part of corncob, flower pesticide position accounted for grain husk shell 1/2 takes fringe for standard, takes after fringe with 70% ethanol surface sterilization, so
Encase whole corncob with plastic sheeting afterwards, keep corncob to moisten and put in refrigerator, Cold pretreatment 3 days at 4 DEG C；?
The ethanol of 85% is disinfected 2～10 minutes, with cutting off flower pesticide top after drying, flower pesticide is placed in the first culture medium, enters
Row inducing culture, light culture 2～after 3 days at 22～28 DEG C, it is transferred in the second culture medium, in 25～29 DEG C, photoperiod
Carry out successive transfer culture under 16h7500lx illumination 8h dark, it is thus achieved that anther callus, continue to turn out bloom control, nursery, transplanting
To land for growing field crops, wait corncob Grain Ripening, i.e. obtained rough dwarf disease resistant corn by screening.
Wherein the composition of the first culture medium described in step (4) and (8) is: N6 basal medium 500mg/L, glycine
4.2mg/L, nicotinic acid 11mg/L, agar 2.3g/L；Kinetins KT 15mg/L, sorbitol 0.5g/L, 2-amino-5-carboxyl valeryl
Amine 20mg/L, alanine 10mg/L, gibberellins GA3 0.8mg/L, insulin 12mg/L, folic acid 16mg/L, colchicine
1.9mg/L；PH value is 6.0.
Described in step (4) and (8), the composition of the second culture medium is: MS minimal medium 700mg/L, inositol 50mg/L,
Glycine 6mg/L, vitamin B1 0.4mg/L, isopropyl-β-D-thiogalactoside 20mg/L, vitamin B6 0.5mg/L,
Sucrose 1.5g/L, naphthalene acetic acid NAA 1.3mg/L, biotin 11mg/L, abscisic acid ABA 3.5mg/L, methyl-nitroso-urea
3.5mg/L, Prohexadione calcium 4.2mg/L, hydrolyzed protein 0.7g/L；And the Maize rough dwarf virus Extracted toxin of concentration 30%
Liquid, pH value is 6.0.
Described in step (3), (7) and (8), the preparation method of Maize rough dwarf virus Extracted toxin liquid is: Semen Maydis of catching an illness
Fringe cervical region cut, cleaning surface is placed in moistening culture dish, and light culture under the conditions of 37 DEG C adds and joins the army after mycelia produces
Support agar to continue to cultivate, obtain bacterial strain；Shaking table in inoculation to fluid medium 37 DEG C is cultivated 20 days, sonic oscillation 10
～30 minutes, filter, centrifugation 15 with 1000～2200rpm～20 minutes, the supernatant obtained i.e. Maize rough dwarf virus
Raw toxin.
The breeding method of a kind of anti-rough dwarf disease Semen Maydis that the present invention relates to, exclusively penetrates into anti-rough dwarf disease gene efficient
Corn breeding trunk strain material, cultivate a kind of anti-rough dwarf disease corn strain, it is to avoid prior art utilizes chemical agent to prevent
Control the environmental pollution that the anti-rough dwarf disease of Semen Maydis causes, shorten breeding cycle, it is to avoid the cytometaplasia that channel genes causes, maintain
Adding its anti-rough dwarf disease ability while breeding excellent properties, decrease Maize Production cost to a certain extent, having should
Use prospect.
Detailed description of the invention
Below by specific embodiment, further technical scheme is specifically described.It should be understood that below
Embodiment be intended only as illustrating, and do not limit the scope of the invention, those skilled in the art is according to the present invention simultaneously
Within the obvious change made and modification are also contained in the scope of the invention.
Embodiment 1
First rough dwarf disease resistant corn material selects to step on sea 3622, and corn breeding trunk strain material selects Zheng Dan 958, institute
State the second rough dwarf disease resistant corn material for gold sea 5.The technology path using the present invention carries out rough dwarf disease breeding for disease resistance, success
Obtain 6 flower training familys that economical character is excellent, rough dwarf disease resistance obtains improvement.
Zheng Dan 958 is the corn variety that China's cultivated area is maximum, has high yield, stable yields advantage, but relatively feels rough dwarf disease.Step on sea
3622 is the kind that Shandong Denghai Seeds Co., Ltd is bred as, high anti-rough dwarf disease, but sense short mosaic disease and Pyrausta nubilalis (Hubern)..Jin Hai
No. 5 is the kind that Chinese farmers breeding expert Zhai Yanju is bred as, high anti-rough dwarf disease, but sense curving cascade and head smut.
Its concrete operations flow process is as follows:
(1) first sowing and the program of transplanting:
Soaking seed respectively in the seed-soaking liquid of dilution by stepping on sea 3622 and Jin Hai 5, after soaking seed 1～2 day, accelerating germination also presses kind
Being seeded in experiment rice seedling bed, individual plant is transplanted, and seeding row spacing is 30 × 30cm；General land for growing field crops rich water and disease pest and weed management, to plant just
It is frequently grown blooming stage, carries out step (2).
Described seed-soaking liquid is 2mL10% dithiocyano-methane cream, 2g prochloraz, 15mL40% good fortune gram suspended seed coating
Agent, 2mL70% imidacloprid water dispersible granule, add 500 times of water dilutions formulated.
(2) acquisition of Hybrid F1:
The corncob of maternal plant is immersed in the warm water of 42 DEG C, take out after soaking 7 minutes, natural cooling 30 minutes, use
The corncob of maternal plant is packaged and marks by paper bag, stand-by.When paternal plant enters florescence, by beautiful on paternal plant
The flower pesticide of meter Sui is uniformly fallen in paper bag on the stigma of maternal plant corncob, and paper bag is continued to package after completing by artificial pollination,
Wait the Grain Ripening of hybrid maize fringe, i.e. obtain Hybrid F1.
(3) the small brown rice planthopper screening of the anti-rough dwarf disease of Hybrid F1:
Hybrid F1 positive season is sowed, when Hybrid F1 was in for 2～2.5 leaf phase, by every strain Semen Maydis 10～the quantity of 20
Discharge small brown rice planthopper to rice seedling bed, after 50 days, every strain Semen Maydis is estimated, retain the plant not encroached on by rough dwarf disease.
Described small brown rice planthopper every feeds Maize rough dwarf virus Extracted toxin liquid 4～5 days, all carries MRDV
Poison.
(4) the antherderived callus induction of Hybrid F1:
What in the middle part of corncob, flower pesticide position accounted for grain husk shell 1/2 takes fringe for standard, takes after fringe with 70% ethanol surface sterilization, so
Encase whole corncob with plastic sheeting afterwards, keep corncob to moisten and put in refrigerator, Cold pretreatment 3 days at 4 DEG C；?
The ethanol of 85% is disinfected 2～10 minutes, with cutting off flower pesticide top after drying, flower pesticide is placed in the first culture medium, enters
Row inducing culture, light culture 2～after 3 days at 22～28 DEG C, it is transferred in the second culture medium, in 25～29 DEG C, photoperiod
Successive transfer culture is carried out under 16h7500lx illumination 8h dark, by the anther callus of the Hybrid F1 of acquisition after 50 days, statistics flower
Medicine callus induction rate, if induction of anther callus rate is less than 5%, then needs Hybrid F1 and rough dwarf disease resistant corn material
Material parent backcrosses until induction of anther callus rate is higher than 5%.
(5) second sowings and the program of transplanting:
Zheng Dan 958 and Hybrid F1 being soaked seed respectively in the seed-soaking liquid of dilution, after soaking seed 1-2 days, accelerating germination also presses kind
Being seeded in experiment rice seedling bed, individual plant is transplanted, and seeding row spacing is 30 × 30cm；General land for growing field crops rich water and disease pest and weed management, to plant just
It is frequently grown blooming stage, carries out step (6).
The same step of described seed-soaking liquid (1).
(6) three acquisitions handing over F1 cenospecies:
The corncob of maternal plant is immersed in the warm water of 42 DEG C, take out after soaking 7 minutes, natural cooling 30 minutes, cut
Remove l/3 grain husk shell and do not open the little Hua of grain husk, with paper bag the corncob of maternal plant packaged and mark, stand-by.Work as paternal plant
When entering florescence, the flower pesticide of corncob on paternal plant is uniformly fallen in paper bag on the stigma of maternal plant corncob, people
Paper bag is continued to package after having pollinated by work, waits the Grain Ripening of hybrid maize fringe, i.e. obtains three friendship F1 cenospecies.
(7) the three small brown rice planthopper screenings handing over the anti-rough dwarf disease of F1 cenospecies:
Three hand over the sowing of F1 cenospecies positive season, when three hand over F1 cenospecies to be in for 2～2.5 leaf phase, by every strain Semen Maydis 10～20
The quantity of head discharges small brown rice planthopper to rice seedling bed, after 50 days is estimated every strain Semen Maydis, retains the plant not encroached on by rough dwarf disease.
Described small brown rice planthopper every feeds Maize rough dwarf virus Extracted toxin liquid 4～5 days, all carries MRDV
Poison.
(8) the three antherderived callus inductions handing over F1 cenospecies:
What in the middle part of corncob, flower pesticide position accounted for grain husk shell 1/2 takes fringe for standard, takes after fringe with 70% ethanol surface sterilization, so
Encase whole corncob with plastic sheeting afterwards, keep corncob to moisten and put in refrigerator, Cold pretreatment 3 days at 4 DEG C；?
The ethanol of 85% is disinfected 2～10 minutes, with cutting off flower pesticide top after drying, flower pesticide is placed in the first culture medium, enters
Row inducing culture, light culture 2～after 3 days at 22～28 DEG C, it is transferred in the second culture medium, in 25～29 DEG C, photoperiod
Carry out successive transfer culture under 16h7500lx illumination 8h dark, it is thus achieved that anther callus, continue to turn out bloom control, nursery, transplanting
To land for growing field crops, wait corncob Grain Ripening, i.e. obtain rough dwarf disease resistant corn.
Wherein the composition of the first culture medium described in step (4) and (8) is: N6 basal medium 500mg/L, glycine
4.2mg/L, nicotinic acid 11mg/L, agar 2.3g/L；Kinetins KT 15mg/L, sorbitol 0.5g/L, 2-amino-5-carboxyl valeryl
Amine 20mg/L, alanine 10mg/L, gibberellins GA3 0.8mg/L, insulin 12mg/L, folic acid 16mg/L, colchicine
1.9mg/L；PH value is 6.0.
Described in step (4) and (8), the composition of the second culture medium is: MS minimal medium 700mg/L, inositol 50mg/L,
Glycine 6mg/L, vitamin B1 0.4mg/L, isopropyl-β-D-thiogalactoside 20mg/L, vitamin B6 0.5mg/L,
Sucrose 1.5g/L, naphthalene acetic acid NAA 1.3mg/L, biotin 11mg/L, abscisic acid ABA 3.5mg/L, methyl-nitroso-urea
3.5mg/L, Prohexadione calcium 4.2mg/L, hydrolyzed protein 0.7g/L；And the Maize rough dwarf virus Extracted toxin of concentration 30%
Liquid, pH value is 6.0.
Described in step (3), (7) and (8), the preparation method of Maize rough dwarf virus Extracted toxin liquid is: Semen Maydis of catching an illness
Fringe cervical region cut, cleaning surface is placed in moistening culture dish, and light culture under the conditions of 37 DEG C adds and joins the army after mycelia produces
Support agar to continue to cultivate, obtain bacterial strain；Shaking table in inoculation to fluid medium 37 DEG C is cultivated 20 days, sonic oscillation 10
～30 minutes, filter, centrifugation 15 with 1000～2200rpm～20 minutes, the supernatant obtained i.e. Maize rough dwarf virus
Raw toxin.
Claims (8)
1. the breeding method of an anti-rough dwarf disease Semen Maydis, it is characterised in that: the breeding method of described anti-rough dwarf disease Semen Maydis is required plants
Sub-material includes, the first rough dwarf disease resistant corn material, the second rough dwarf disease resistant corn material and corn breeding trunk strain
Material；
Described corn breeding trunk strain material is yield height, meter Zhi Hao, the germline of susceptible rough dwarf disease, and described first rough dwarf disease resists
Property corn material and the second rough dwarf disease resistant corn material are high anti-rough dwarf disease, not high yield, are at source not suitable for large-scale promotion
Germline；
Specifically include following steps:
(1) first sowing and the program of transplanting:
First rough dwarf disease resistant corn material and the second rough dwarf disease resistant corn material are soaked seed respectively in the seed-soaking liquid of dilution,
After soaking seed 1～2 day, accelerating germination is also seeded in experiment rice seedling bed by kind, and individual plant is transplanted, and seeding row spacing is 30 × 30cm；General land for growing field crops is fertile
Water and disease pest and weed management, to normal plants to blooming stage, carry out step (2)；
(2) acquisition of Hybrid F1:
The corncob of maternal plant is immersed in the warm water of 39～46 DEG C, take out after soaking 6～10 minutes, natural cooling 30 points
Clock, packages the corncob of maternal plant with paper bag and marks, stand-by；When paternal plant enters florescence, male parent is planted
In strain, the flower pesticide of corncob is uniformly fallen in paper bag on the stigma of maternal plant corncob, and paper bag is continued after completing by artificial pollination
Package, wait the Grain Ripening of hybrid maize fringe, i.e. obtain Hybrid F1；
(3) the small brown rice planthopper screening of the anti-rough dwarf disease of Hybrid F1:
Hybrid F1 positive season is sowed, when Hybrid F1 was in for 2～2.5 leaf phase, by the quantity of every strain Semen Maydis 10～20 to seedling
Field release small brown rice planthopper, is estimated every strain Semen Maydis after 50 days, retains the plant not encroached on by rough dwarf disease；
Described small brown rice planthopper every feeds Maize rough dwarf virus Extracted toxin liquid 4～5 days, all carries Maize rough dwarf virus；
(4) the antherderived callus induction of Hybrid F1:
What in the middle part of corncob, flower pesticide position accounted for grain husk shell 1/2 takes fringe for standard, takes after fringe with 70% ethanol surface sterilization, then uses
Plastic sheeting encases whole corncob, keeps corncob to moisten and puts in refrigerator, Cold pretreatment 3 days at 4 DEG C；85%
Ethanol is disinfected 2～10 minutes, with cutting off flower pesticide top after drying, flower pesticide is placed in the first culture medium, induces
Cultivate, light culture 2～after 3 days at 22～28 DEG C, be transferred in the second culture medium, in 25～29 DEG C, photoperiod 16h7500lx
Carry out successive transfer culture under illumination 8h dark, by the anther callus of the Hybrid F1 of acquisition after 50 days, add up antherderived callus group
Knit inductivity, if induction of anther callus rate is less than 5%, then need to return Hybrid F1 with rough dwarf disease resistant corn material parent
Hand over until induction of anther callus rate is higher than 5%；
(5) second sowings and the program of transplanting:
Corn breeding trunk strain material and Hybrid F1 are soaked seed respectively in the seed-soaking liquid of dilution, after soaking seed 1-2 days, accelerating germination
And it being seeded in experiment rice seedling bed by kind, individual plant is transplanted, and seeding row spacing is 30 × 30cm；General land for growing field crops rich water and disease pest and weed management,
To normal plants to blooming stage, carry out step (6)；The same step of described seed-soaking liquid (1)；
(6) three acquisitions handing over F1 cenospecies:
The corncob of maternal plant is immersed in the warm water of 39～46 DEG C, take out after soaking 6～10 minutes, natural cooling 30 points
Clock, cuts off l/3 grain husk shell and does not opens the little Hua of grain husk, packaged by the corncob of maternal plant with paper bag and mark, stand-by；Work as male parent
When plant enters florescence, the flower pesticide of corncob on paternal plant is uniformly fallen the stigma of maternal plant corncob in paper bag
On, paper bag is continued to package after completing by artificial pollination, waits the Grain Ripening of hybrid maize fringe, i.e. obtains three friendship F1 cenospecies；
(7) the three small brown rice planthopper screenings handing over the anti-rough dwarf disease of F1 cenospecies:
Three hand over the sowing of F1 cenospecies positive seasons, when three hand over F1 cenospecies to be in for 2～2.5 leaf phase, by every strain Semen Maydis 10～20
Quantity discharges small brown rice planthopper to rice seedling bed, after 50 days is estimated every strain Semen Maydis, retains the plant not encroached on by rough dwarf disease；
Described small brown rice planthopper every feeds Maize rough dwarf virus Extracted toxin liquid 4～5 days, all carries Maize rough dwarf virus；
(8) the three antherderived callus inductions handing over F1 cenospecies:
What in the middle part of corncob, flower pesticide position accounted for grain husk shell 1/2 takes fringe for standard, takes after fringe with 70% ethanol surface sterilization, then uses
Plastic sheeting encases whole corncob, keeps corncob to moisten and puts in refrigerator, Cold pretreatment 3 days at 4 DEG C；85%
Ethanol is disinfected 2～10 minutes, with cutting off flower pesticide top after drying, flower pesticide is placed in the first culture medium, induces
Cultivate, light culture 2～after 3 days at 22～28 DEG C, be transferred in the second culture medium, in 25～29 DEG C, photoperiod 16h7500lx
Carry out successive transfer culture under illumination 8h dark, it is thus achieved that anther callus, continue to turn out bloom control, nursery, be transplanted to land for growing field crops, etc.
Treat corncob Grain Ripening, i.e. obtained rough dwarf disease resistant corn by screening.
The breeding method of a kind of anti-rough dwarf disease Semen Maydis the most according to claim 1, it is characterised in that: described corn breeding master
Dry product based material is Zheng Dan 958, and described first rough dwarf disease resistant corn material is for stepping on sea 3622, and described second rough dwarf disease resistance is beautiful
Rice material is gold sea 5.
The breeding method of a kind of anti-rough dwarf disease Semen Maydis the most according to claim 1, it is characterised in that: step (1) described leaching
Planting liquid is 2mL10% dithiocyano-methane cream, 2g prochloraz, 15mL40% good fortune gram suspension seed-coating agent, 2mL70% imidacloprid
Water dispersible granules, adds 500 times of water dilutions formulated.
The breeding method of a kind of anti-rough dwarf disease Semen Maydis the most according to claim 1, it is characterised in that: step (2) or step
(6) warm water temperature that the spike of rice of described maternal plant immerses is 42 DEG C, and described soak time is 7 minutes.
5. according to the breeding method of a kind of anti-rough dwarf disease Semen Maydis described in any one of Claims 1 to 4, it is characterised in that: wherein
Described in step (4) or (8), the composition of the first culture medium is: N6 basal medium 500mg/L, glycine 4.2mg/L, nicotinic acid
11mg/L, agar 2.3g/L；Kinetins KT 15mg/L, sorbitol 0.5g/L, 2-amino-5-carboxyl pentanamide 20mg/L, the third ammonia
Acid 10mg/L, gibberellins GA3 0.8mg/L, insulin 12mg/L, folic acid 16mg/L, colchicine 1.9mg/L；PH value is 6.0.
6. according to the breeding method of a kind of anti-rough dwarf disease Semen Maydis described in any one of Claims 1 to 4, it is characterised in that: step
(4) or the composition of the second culture medium described in (8) is: MS minimal medium 700mg/L, inositol 50mg/L, glycine 6mg/L,
Vitamin B1 0.4mg/L, isopropyl-β-D-thiogalactoside 20mg/L, vitamin B6 0.5mg/L, sucrose 1.5g/L, naphthalene
Acetic acid NAA 1.3mg/L, biotin 11mg/L, abscisic acid ABA3.5mg/L, methyl-nitroso-urea 3.5mg/L, Prohexadione calcium
4.2mg/L, hydrolyzed protein 0.7g/L；And the Maize rough dwarf virus Extracted toxin liquid of concentration 30%, pH value is 6.0.
7. according to the breeding method of a kind of anti-rough dwarf disease Semen Maydis described in any one of Claims 1 to 4, it is characterised in that: step
(3) and the preparation method of (7) described Characters of Erwinia chrysanthemi pv. zeae bacterium Extracted toxin liquid is: the fringe cervical region of Semen Maydis of catching an illness is cut
Under, cleaning surface is placed in moistening culture dish, light culture under the conditions of 37 DEG C, adds Nutrient agar and continue after mycelia produces
Cultivate, obtain bacterial strain；Shaking table in inoculation to fluid medium 37 DEG C is cultivated 20 days, sonic oscillation 10～30 minutes,
Filter, centrifugation 15 with 1000～2200rpm～20 minutes, the supernatant obtained i.e. Maize rough dwarf virus Raw toxin.
The breeding method of a kind of anti-rough dwarf disease Semen Maydis the most according to claim 6, it is characterised in that: step (8) described water
The preparation method of rice bacterial Footrot bacterium Extracted toxin liquid is: cut by the fringe cervical region of Semen Maydis of catching an illness, and cleaning surface is rearmounted
In moistening culture dish, light culture under the conditions of 37 DEG C, after mycelia produces, add Nutrient agar continue to cultivate, obtain bacterial strain；
Shaking table in inoculation to fluid medium 37 DEG C is cultivated 20 days, sonic oscillation 10～30 minutes, filter, with 1000～
The centrifugation 15 of 2200rpm～20 minutes, the supernatant obtained i.e. Maize rough dwarf virus Raw toxin.
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