CN105900828A

CN105900828A - Woad-cabbage distant hybridization tetraploid breeding method

Info

Publication number: CN105900828A
Application number: CN201610236279.2A
Authority: CN
Inventors: 周常
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-03-15
Filing date: 2016-04-14
Publication date: 2016-08-31

Abstract

The invention discloses a woad-cabbage distant hybridization tetraploid breeding method. The method comprises the following steps: germinating seeds at low temperature; carrying out light culture on sprouts under microgravity condition to grow seedlings, and transplanting two kinds of seedlings into the field until blossoming; germinating seeds, and inducing chromosome doubling by synergism of colchicines and alpha-ray irradiation; screening tetraploid sprouts, continuously growing, transplanting the obtained tetraploid plantlets into the field for culture, carrying out self-pollination to obtain woad-cabbage distant hybridization tetraploid seeds, and continuously sowing Generation 5-Genaration 8 so as to obtain woad-cabbage distant hybridization tetraploid seeds with stable characters. By using woad as female parent and cabbage as male parent for distant hybridization breeding, progeny hybrids with various excellent characters are formed, and species characters are richer and various.

Description

A kind of woaded blue-wild cabbage distant hybridization tetraploid breeding method

Technical field

The present invention relates to the breeding field of woaded blue, be specifically related to a kind of woaded blue-wild cabbage distant hybridization tetraploid breeding Method.

Background technology

Woaded blue is that a kind of Cruciferae woaded blue belongs to biennial herb plant, and the Origin of Species, in China, has the strongest Cold-resistant drought-enduring adaptation ability, extensive growth is distributed in China various places.Woaded blue root can be made after shining dry-cure For Radix Isatidis, its leaf is the most then Chinese medicine folium isatidis.This two tastes medicinal material have clearing heat and detoxicating, The function of cooling blood and relieving pain.Seeds of Isatis indigotica can be as oil expression, as the raw material sources of industry oil, and woaded blue Indigo dye can also be extracted in leaf position, for people to use.Visible woaded blue have outstanding use value and Economic worth.

Wild cabbage is a kind of Cruciferae Brassica genus biennial herb plant, and the Origin of Species is along the line in Mediterranean, tool Having preferable cold tolerance, adaptive faculty is strong, and before 4000, people just have begun to plantation.In China Extensively plant in the north and south China and other places, be a kind of important vegetable.Wild cabbage also has certain nourishing the stomach and is good for The pharmacology function of kidney, it is possible to protection renovating tummy mucous membrane, is the good source of potassium element.Additionally, wild cabbage is also Show certain ornamental value.

Distant hybridization refers to the hybridization carried out between the species of different genera, can be species between different genera Proterties is combined reconstruction, produces new variation proterties, thus constructs the species made new advances.It is said that in general, it is remote Edge crossbreeding easily occurs that offspring early dies young and sterile problem, and the relationship between the two of distant hybridization species Relation is the nearest, is hybridized to power the highest, and filial generation existence reproduction ability is the strongest.And added by chromosome Times technology, the polyploid of generation makes the chromosome nonhomologous of normal reductive division to recover normally to breed.

Chinese patent CN200810301666.5 discloses the breeding method of a kind of plant strain of distant hybrid lily, Ovary and flower pesticide are carried out cultured in vitro, then use with the unconventional Pollination Modes that ovary is cut off carry out from Body pollinate, hereafter carry out embryo culture hybrid plant.But it is maternal to there is male parent in the cenospecies of this patent In crossover process, chromosome does not mates, it is impossible to paired phenomenon, and the seed height causing hybridization to obtain is sterile, Can not obtain the seed of same proterties continually and steadily, social value and economic worth are low.

Chinese patent CN201410175221.2 discloses a kind of by the rescue isolated cultivation acquisition remote edge of strawberry The method of hybrid and culture medium, this invention Charlie the most fragrant with planting strawberry kind, sweet, Feng Xiang are for maternal or father This, wild proso millet prolamin is male parent or female parent hybridizes, and is just giving reciprocal cross to carry out, by hybridization simultaneously Immature embryo after pollination carries out cultured in vitro, is drawn off carrying out Rescue culture before aborted embryo, it is thus achieved that remote Edge filial generation.But parent's high number needed for this patent, needs to expend a large amount of manpower and materials, and only enters Row rescue culture means are difficult to overcome the low success rate of problem of distant hybridization, there is filial generation the most sterile simultaneously Problem.

By analyzing it can be seen that brassica plant and woaded blue belong to, to be likely to be formed proterties between woaded blue the most remote Edge Hybrid, belongs to woaded blue crossbreeding about Brassica genus wild cabbage and woaded blue and then reports seldom, therefore this The bright cross breeding method being intended to be provided woaded blue and wild cabbage by distant hybridization, tetraploid breeding technology.

Summary of the invention

The present invention is directed to the problems referred to above, it is provided that a kind of woaded blue-wild cabbage distant hybridization tetraploid breeding method.

The present invention solves the problems referred to above and be the technical scheme is that

A kind of woaded blue-wild cabbage distant hybridization tetraploid breeding method, comprises the following steps:

Step one, low temperature rudiment: select to plant the full Seeds of Isatis indigotica of grain and cabbage seed, molten at formalin Soaking 30min～50min in liquid, rub removal seed coat, the seed after then processing wraps up with wet cloth, Under the temperature conditions of 3 DEG C～8 DEG C, carrying out vernalization processing 48h～72h, until growing the woaded blue of 3mm～8mm Young shoot and wild cabbage young shoot；

Step 2, illumination cultivation: by woaded blue young shoot and wild cabbage young shoot culture transferring respectively to fluid nutrient medium, make Culture medium the most at the uniform velocity rotates, and uses xenon lamp illumination simulation to cultivate 2 days～3 days, until growing up to The woaded blue seedling of a length of 10cm～15cm of strain and cabbage seedling, respectively by two kinds of seedling replantings to big Tanaka, Giving normal illumination, moisture and fertilizer condition, the farm work carrying out necessity is safeguarded, until two kinds of plants Bloom；

Step 3, distant hybridization: the woaded blue obtained in step 2 is as female parent, and wild cabbage is male parent, use thin Hairbrush careful collection wild cabbage male flower pollen, adds water and makes pollen suspension, uses little banister brush to be suspended by pollen Liquid spreads upon on woaded blue female flower style column cap, smears acetone-ethanol more molten on woaded blue female flower style column cap Liquid, promotes cross-pollination process, carries out bagging process, tighten sack, prevent external source pollen after having pollinated Interference, then proceedes to cultivate, obtains woaded blue-wild cabbage crossbred plant, gathers in the crops seed, standby；

Step 4, tetraploid breeding: seed step 3 obtained carries out sprouting to be cultivated, and bears young shoot, makes With the every 4h uniform application of the colchicin that concentration is 2%～20% on young shoot, continue 100h～160h；Afterwards Use alpha ray irradiation 20h～40h, the young shoot processed use flow cytometer is carried out ploidy analysis, sieve Select tetraploid young shoot, the tetraploid young shoot filtered out is transplanted to continued growth in B5 medium, until The tetraploid plant of a length of 5cm～8cm of long strain；

Step 5, the production of hybrid seeds: tetraploid plant step 4 obtained moves to big Tanaka and cultivates, self-pollination, Collect after plant maturation and obtain woaded blue-wild cabbage distant hybridization tetraploid seed, continuously 5 generations of sowing～8 generations, To woaded blue-wild cabbage distant hybridization tetraploid seed that proterties is stable, to obtain final product.

Further, in step one, the concentration of formalin solution is 2%～10%.

Further, in step 2, the composition of fluid nutrient medium and weight portion thereof be: 100 parts of ammonium nitrate～300 Part, 150 parts～250 parts of potassium chloride, 80 parts～150 parts of ammonium hydrogen carbonate, potassium dihydrogen phosphate 150 parts～280 Part, KI 30 parts～80 parts, sodium iodide 30 parts～60 parts, 20 parts～50 parts of calcium chloride, magnesium metaborate 50 parts～100 parts, 30 parts～80 parts of ferrous sulfate, frerrous chloride 30 parts～80 parts, manganese sulfate 20 parts～50 Part, zinc gluconate 20 parts～60 parts, calcium gluconae 20 parts～50 parts, sodium potassium tartrate tetrahydrate 20 parts～40 Part, aspartic acid 20 parts～50 parts, glycine 20 parts～60 parts, phenylalanine 20 parts～40 parts, nicotinoyl Amine 20 parts～60 parts, L-AA 20 parts～50 parts, galactolipin 3000 parts～5000 parts, glucose 3000 Part～5000 parts, 2000 parts～4000 parts of agar, 5000 parts～10000 parts of water.

Further, in step 2, the field intensity of microgravity is 0.01g～0.15g.

Further, in step 2, the rotating speed at the uniform velocity rotated is 35r/min～70r/min.

Further, in step 3, in acetone-ethanol solution, acetone is 1:20～30 with the volume ratio of ethanol.

Further, in step 4, the close rate in alpha ray source be 3500 roentgens/hour～9000 roentgens/little Time.

The invention have the advantage that

1. by carrying out distant hybridization method breeding for maternal wild cabbage for male parent with woaded blue, by introducing external source Property gene method make original genome type rebuild, easily form various proterties excellent Offspring hybrid, make species proterties more rich and varied；

2. use tetraploid breeding method make the technique improvement of cenospecies chromosome doubling and recovered offspring The survival ability of hybrid plant and breed level, makes the outstanding proterties of sudden change be continued heredity, And by doubling the many times of chromosomes formed, its plant has that branches and leaves are huge, rhizome is healthy and strong Desirable features, makes various effective pharmaceutical ingredient content be improved；

3. the cost advantages such as use wild cabbage is as parent sire of hybrid pigs, has raw material sources extensive, cheap and easy to get, And wild cabbage planting range is big, species proterties is many, bring abundant change easily to advanced generation cross kind Change potentiality.

Detailed description of the invention

Embodiment 1

Comprise the following steps:

Step one, low temperature rudiment: select to plant the full Seeds of Isatis indigotica of grain and cabbage seed, be 2% in concentration Formalin solution in soak 30min, rub removal seed coat, the seed after then processing is with wet Cloth wrap up, carry out under the temperature conditions of 3 DEG C vernalization process 48h, until grow 3mm woaded blue young shoot and Wild cabbage young shoot.

Step 2, illumination cultivation: by woaded blue young shoot and wild cabbage young shoot culture transferring respectively to fluid nutrient medium, make Culture medium is 0.01g in microgravity field intensity, and rotating speed is at the uniform velocity to rotate under conditions of 35r/min, uses xenon lamp mould Intend illumination cultivation 2 days, until the woaded blue seedling of a length of 10cm of long strain and cabbage seedling, respectively by two kinds Seedling replanting, to big Tanaka, gives normal illumination, moisture and fertilizer condition, carries out the farm work of necessity Safeguard, until two kinds of flowering of plant.Wherein composition and the weight portion thereof of fluid nutrient medium is: ammonium nitrate 100 Part, 150 parts of potassium chloride, 80 parts of ammonium hydrogen carbonate, potassium dihydrogen phosphate 150 parts, KI 30 parts, iodate 30 parts of sodium, 20 parts of calcium chloride, magnesium metaborate 50 parts, 30 parts of ferrous sulfate, frerrous chloride 30 parts, Manganese sulfate 20 parts, zinc gluconate 20 parts, calcium gluconae 20 parts, sodium potassium tartrate tetrahydrate 20 parts, asparagus fern Propylhomoserin 20 parts, glycine 20 parts, phenylalanine 20 parts, niacinamide 20 parts, L-AA 20 parts, Galactolipin 3000 parts, glucose 3000 parts, 2000 parts of agar, 5000 parts of water.

Step 3, distant hybridization: the woaded blue obtained in step 2 is as female parent, and wild cabbage is male parent, use thin Hairbrush careful collection wild cabbage male flower pollen, adds water and makes pollen suspension, uses little banister brush to be suspended by pollen Liquid spreads upon on woaded blue female flower style column cap, smears acetone-ethanol more molten on woaded blue female flower style column cap Liquid, promotes cross-pollination process, carries out bagging process, tighten sack, prevent external source pollen after having pollinated Interference, then proceedes to cultivate, obtains woaded blue-wild cabbage crossbred plant, gathers in the crops seed, standby.Wherein acetone In-ethanol solution, acetone is 1:20 with the volume ratio of ethanol.

Step 4, tetraploid breeding: seed step 3 obtained carries out sprouting to be cultivated, and bears young shoot, makes With the every 4h uniform application of the colchicin that concentration is 2% on young shoot, continue 100h；Dosage rate afterwards Be 3500 roentgens/hour alpha ray irradiation 20h, by the young shoot processed use flow cytometer carry out again Property analyze, filter out tetraploid young shoot, the tetraploid young shoot that filters out transplanted and continues to B5 medium Growth, until the tetraploid plant of a length of 5cm of long strain.

Step 5, the production of hybrid seeds: tetraploid plant step 4 obtained moves to big Tanaka and cultivates, self-pollination, Collect after plant maturation and obtain woaded blue-wild cabbage distant hybridization tetraploid seed, continuously 5 generations of sowing, obtain proterties Stable woaded blue-wild cabbage distant hybridization tetraploid seed, to obtain final product.

Embodiment 2

Comprise the following steps:

Step one, low temperature rudiment: select to plant the full Seeds of Isatis indigotica of grain and cabbage seed, be 10% in concentration Formalin solution in soak 50min, rub removal seed coat, the seed after then processing is with wet Cloth wrap up, carry out under the temperature conditions of 8 DEG C vernalization process 72h, until grow 8mm woaded blue young shoot and Wild cabbage young shoot.

Step 2, illumination cultivation: by woaded blue young shoot and wild cabbage young shoot culture transferring respectively to fluid nutrient medium, make Culture medium is 0.15g in microgravity field intensity, and rotating speed is at the uniform velocity to rotate under conditions of 70r/min, uses xenon lamp mould Intend illumination cultivation 3 days, until the woaded blue seedling of a length of 15cm of long strain and cabbage seedling, respectively by two kinds Seedling replanting, to big Tanaka, gives normal illumination, moisture and fertilizer condition, carries out the farm work of necessity Safeguard, until two kinds of flowering of plant.Wherein composition and the weight portion thereof of fluid nutrient medium is: ammonium nitrate 300 Part, 250 parts of potassium chloride, 150 parts of ammonium hydrogen carbonate, potassium dihydrogen phosphate 280 parts, KI 80 parts, iodine 60 parts of sodium of change, 50 parts of calcium chloride, magnesium metaborate 100 parts, 80 parts of ferrous sulfate, frerrous chloride 80 parts, Manganese sulfate 50 parts, zinc gluconate 60 parts, calcium gluconae 50 parts, sodium potassium tartrate tetrahydrate 40 parts, asparagus fern Propylhomoserin 50 parts, glycine 60 parts, phenylalanine 40 parts, niacinamide 60 parts, L-AA 50 parts, Galactolipin 5000 parts, glucose 5000 parts, 4000 parts of agar, 10000 parts of water.

Step 3, distant hybridization: the woaded blue obtained in step 2 is as female parent, and wild cabbage is male parent, use thin Hairbrush careful collection wild cabbage male flower pollen, adds water and makes pollen suspension, uses little banister brush to be suspended by pollen Liquid spreads upon on woaded blue female flower style column cap, smears acetone-ethanol more molten on woaded blue female flower style column cap Liquid, promotes cross-pollination process, carries out bagging process, tighten sack, prevent external source pollen after having pollinated Interference, then proceedes to cultivate, obtains woaded blue-wild cabbage crossbred plant, gathers in the crops seed, standby.Wherein acetone In-ethanol solution, acetone is 1:30 with the volume ratio of ethanol.

Step 4, tetraploid breeding: seed step 3 obtained carries out sprouting to be cultivated, and bears young shoot, makes With the every 4h uniform application of the colchicin that concentration is 20% on young shoot, continue 160h；Dosage afterwards Rate be 9000 roentgens/hour alpha ray irradiation 40h, by the young shoot processed use flow cytometer carry out Ploidy analysis, filters out tetraploid young shoot, transplants the tetraploid young shoot filtered out to B5 medium relaying Continuous growth, until the tetraploid plant of a length of 8cm of long strain.

Step 5, the production of hybrid seeds: tetraploid plant step 4 obtained moves to big Tanaka and cultivates, self-pollination, Collect after plant maturation and obtain woaded blue-wild cabbage distant hybridization tetraploid seed, continuously 8 generations of sowing, obtain proterties Stable woaded blue-wild cabbage distant hybridization tetraploid seed, to obtain final product.

Embodiment 3

Comprise the following steps:

Step one, low temperature rudiment: select to plant the full Seeds of Isatis indigotica of grain and cabbage seed, be 6% in concentration Formalin solution in soak 40min, rub removal seed coat, the seed after then processing is with wet Cloth wrap up, carry out under the temperature conditions of 6 DEG C vernalization process 60h, until grow 6mm woaded blue young shoot and Wild cabbage young shoot.

Step 2, illumination cultivation: by woaded blue young shoot and wild cabbage young shoot culture transferring respectively to fluid nutrient medium, make Culture medium is 0.08g in microgravity field intensity, and rotating speed is at the uniform velocity to rotate under conditions of 57r/min, uses xenon lamp mould Intend illumination cultivation 3 days, until the woaded blue seedling of a length of 13cm of long strain and cabbage seedling, respectively by two kinds Seedling replanting, to big Tanaka, gives normal illumination, moisture and fertilizer condition, carries out the farm work of necessity Safeguard, until two kinds of flowering of plant.Wherein composition and the weight portion thereof of fluid nutrient medium is: ammonium nitrate 200 Part, 200 parts of potassium chloride, 115 parts of ammonium hydrogen carbonate, potassium dihydrogen phosphate 215 parts, KI 55 parts, iodine 45 parts of sodium of change, 35 parts of calcium chloride, magnesium metaborate 75 parts, 55 parts of ferrous sulfate, frerrous chloride 55 parts, Manganese sulfate 35 parts, zinc gluconate 40 parts, calcium gluconae 35 parts, sodium potassium tartrate tetrahydrate 30 parts, asparagus fern Propylhomoserin 35 parts, glycine 40 parts, phenylalanine-3,4-quinone 0 part, niacinamide 40 parts, L-AA 35 parts, Galactolipin 4000 parts, glucose 4000 parts, 3000 parts of agar, 7500 parts of water.

Step 3, distant hybridization: the woaded blue obtained in step 2 is as female parent, and wild cabbage is male parent, use thin Hairbrush careful collection wild cabbage male flower pollen, adds water and makes pollen suspension, uses little banister brush to be suspended by pollen Liquid spreads upon on woaded blue female flower style column cap, smears acetone-ethanol more molten on woaded blue female flower style column cap Liquid, promotes cross-pollination process, carries out bagging process, tighten sack, prevent external source pollen after having pollinated Interference, then proceedes to cultivate, obtains woaded blue-wild cabbage crossbred plant, gathers in the crops seed, standby.Wherein acetone In-ethanol solution, acetone is 1:25 with the volume ratio of ethanol.

Step 4, tetraploid breeding: seed step 3 obtained carries out sprouting to be cultivated, and bears young shoot, makes With the every 4h uniform application of the colchicin that concentration is 11% on young shoot, continue 130h；Dosage afterwards Rate be 6300 roentgens/hour alpha ray irradiation 30h, by the young shoot processed use flow cytometer carry out Ploidy analysis, filters out tetraploid young shoot, transplants the tetraploid young shoot filtered out to B5 medium relaying Continuous growth, until the tetraploid plant of a length of 7cm of long strain.

Step 5, the production of hybrid seeds: tetraploid plant step 4 obtained moves to big Tanaka and cultivates, self-pollination, Collect after plant maturation and obtain woaded blue-wild cabbage distant hybridization tetraploid seed, continuously 7 generations of sowing, obtain proterties Stable woaded blue-wild cabbage distant hybridization tetraploid seed, to obtain final product.

Experimental example

To the hybrid woaded blue prepared according to the breeding method of embodiment 1～3 respectively, be numbered Y1, Y2, Y3, as experimental group；Woaded blue to common growth, is labeled as Y0, as a control group.Right by experiment The breeding effect of experimental group and control group is tested, and test result is shown in Table 1.

Table 1 breeding effect test result

Note: "/" represents non-hybridized.

Conclusion: from comparing result, the breeding of the distant hybridization tetraploid isatis of embodiment 1～3 gained becomes Power is higher, and the hybrid plant obtained by woaded blue-wild cabbage distant hybridization tetraploid breeding method, greatly Width improves the effective component of woaded blue, has the highest medical value and economic worth.

These are only the preferred embodiments of the present invention, be not limited to the present invention, for this area For technical staff, the present invention can have various modifications and variations.All within the spirit and principles in the present invention, Any modification, equivalent substitution and improvement etc. made, should be included within the scope of the present invention.

Claims

1. woaded blue-wild cabbage distant hybridization tetraploid breeding method, it is characterised in that include following step Rapid:

Step one, low temperature rudiment: select to plant the full Seeds of Isatis indigotica of grain and cabbage seed, formal Soaking 30min～50min in woods solution, rub removal seed coat, the seed after then processing is used Wet cloth wraps up, and carries out vernalization and process 48h～72h, until growing under the temperature conditions of 3 DEG C～8 DEG C The woaded blue young shoot of 3mm～8mm and wild cabbage young shoot；

Step 2, illumination cultivation: by woaded blue young shoot and wild cabbage young shoot culture transferring respectively to fluid nutrient medium, Make culture medium the most at the uniform velocity rotate, use xenon lamp illumination simulation cultivation 2 days～3 days, directly To woaded blue seedling and the cabbage seedling of a length of 10cm～15cm of long strain, respectively two kinds of seedling replantings are arrived Big Tanaka, gives normal illumination, moisture and fertilizer condition, and the farm work carrying out necessity is safeguarded, Until two kinds of flowering of plant；

Step 3, distant hybridization: the woaded blue obtained in step 2 is as female parent, and wild cabbage is male parent, makes With fine, soft fur brush careful collection wild cabbage male flower pollen, add water and make pollen suspension, use little banister brush to incite somebody to action Pollen suspension spreads upon on woaded blue female flower style column cap, is coated with on woaded blue female flower style column cap the most again Smear acetone-ethanol solution, promote cross-pollination process, carry out bagging process after having pollinated, tighten bag Mouthful, prevent external source pollen from disturbing, then proceed to cultivate, obtain woaded blue-wild cabbage crossbred plant, results Seed, standby；

Step 4, tetraploid breeding: seed step 3 obtained carries out sprouting to be cultivated, and bears young shoot, Concentration be colchicin every 4h uniform application of 2%～20% on young shoot, continue 100h～160h； Use alpha ray irradiation 20h～40h afterwards, the young shoot processed use flow cytometer is carried out ploidy Analyze, filter out tetraploid young shoot, the tetraploid young shoot filtered out is transplanted to B5 medium relaying Continuous growth, until the tetraploid plant of a length of 5cm～8cm of long strain；

Step 5, the production of hybrid seeds: tetraploid plant step 4 obtained moves to big Tanaka cultivation, awards from flower Powder, collects after plant maturation and obtains woaded blue-wild cabbage distant hybridization tetraploid seed, continuously sowing 5 generations～8 In generation, obtain the stable woaded blue of proterties-wild cabbage distant hybridization tetraploid seed, to obtain final product.

Breeding method the most according to claim 1, it is characterised in that in step one, described formalin is molten The concentration of liquid is 2%～10%.

Breeding method the most according to claim 1, it is characterised in that in step 2, described fluid nutrient medium Composition and weight portion be: 100 parts～300 parts of ammonium nitrate, 150 parts～250 parts of potassium chloride, carbonic acid Hydrogen ammonium 80 parts～150 parts, potassium dihydrogen phosphate 150 parts～280 parts, KI 30 parts～80 parts, iodate 30 parts～60 parts of sodium, 20 parts～50 parts of calcium chloride, magnesium metaborate 50 parts～100 parts, ferrous sulfate 30 Part～80 parts, frerrous chloride 30 parts～80 parts, manganese sulfate 20 parts～50 parts, zinc gluconate 20 parts～60 Part, calcium gluconae 20 parts～50 parts, sodium potassium tartrate tetrahydrate 20 parts～40 parts, aspartic acid 20 parts～50 Part, glycine 20 parts～60 parts, phenylalanine 20 parts～40 parts, niacinamide 20 parts～60 parts, L- 20 parts～50 parts of ascorbic acid, galactolipin 3000 parts～5000 parts, glucose 3000 parts～5000 parts, 2000 parts～4000 parts of agar, 5000 parts～10000 parts of water.

Breeding method the most according to claim 1, it is characterised in that in step 2, the field of described microgravity Strong is 0.01g～0.15g.

Breeding method the most according to claim 1, it is characterised in that in step 2, described at the uniform velocity rotates Rotating speed is 35r/min～70r/min.

Breeding method the most according to claim 1, it is characterised in that in step 3, described acetone-ethanol In solution, acetone is 1:20～30 with the volume ratio of ethanol.

Breeding method the most according to claim 1, it is characterised in that in step 4, described alpha ray source Close rate be 3500 roentgens/hour～9000 roentgens/hour.