CN108617502A - A method of building capsicum mutant library using ethylmethane sulfonate - Google Patents
A method of building capsicum mutant library using ethylmethane sulfonate Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a kind of methods building capsicum mutant library using ethylmethane sulfonate, the present invention is using " abiding by peppery No. 1 " as mutagenesis object, under the premise of pointing out the treatment fluid dosage of every seed and excluding influence of the space to germination percentage, by comparing pepper seed germination rate of the various concentration EMS treatment fluids under different mutation times, determine half lethal dose, half lethal dose mutagenic treatment pepper seed is used in combination, investigates M2The frequency of mutation and mutation type in generation, identification of M4In generation, can stablize the mutant of heredity, build mutant library, the mutation types such as leaf, stem, fruit, breeding time, floral organ, fertility are obtained, have formulated abundant material for capsicum functional genomics research, while part beneficial mutation may be directly applied to breeding practice.
Description
Technical field
The invention belongs to crops mutation breeding technologies fields, are related to a kind of utilization ethylmethane sulfonate structure capsicum mutation
The method in body library.
Background technology
Capsicum is one of most important vegetable crop in China, and cultivated area accounts for about the 10% of the vegetable cultivation gross area,
Ensure to play an important role in China's vegetables anniversary stable market supply.The pepper planting area whole nation first in Guizhou, and the kind of capsicum annum fasciculatum
It plants area and accounts for about the 40% of the whole province's pepper planting gross area, therefore, capsicum annum fasciculatum accounts in the capsicum industry in Guizhou Province or even the whole nation
There is very important status.It will be further appreciated that Guizhou capsicum annum fasciculatum gives off a strong fragrance, moderate, the unique flavor quality of peppery degree
There is not replaceability to Guizhou people.Although the flavor quality and industrial advantage of capsicum annum fasciculatum protrude, still deposit in production practice
In certain deficiency：First, there is the marketing quality that wrinkle reduces fruit in fruit face, second is that Guizhou aphid insect pest weight, capsicum are subject to
Virosis infection and sprawling, third, plant type is loosely unfavorable for rational close planting, affect red pepper high-yield caused by aphid feeding.That
The mutant library based on local capsicum annum fasciculatum kind is built with regard to particularly important.
It is concentrated mainly on including space mutagenesis, gamma-rays, laser currently, obtaining the induced-mutation technique that capsicum mutant is utilized
On the radioinduction including ion beam, and radioinduction easily causes chromosome aberration, injures greatly, causes lethal to chromosome
More, the spectrum of mutation the stability of mutation are low, and various rays must be generated by expensive instrument and equipment, and dosage is not easy to control, behaviour
Make complexity, it is of high cost.
Ethylmethane sulfonate (EMS) is a kind of common chemical mutagen, has easy to operate, at low cost, mutagenic frequency
Height, mutational range is wide, can generate the advantages such as mass mutation body in a short time, have been successfully applied for grain, oil plant, vegetables
Etc. various crops mutant library structure, and functional genome and genetic thremmatology research in achieve significant achievement.But
Ethylmethane sulfonate is less for the breeding research of capsicum, even more the not no relevant report about the mutant library of capsicum annum fasciculatum.
Invention content
A kind of method building capsicum mutant library using ethylmethane sulfonate is educated present invention solves the problem in that providing,
Multiple types mutant can be obtained using this method, abundant material is formulated for capsicum functional genomics research, is sieved simultaneously
It selects advantageous mutation and directly applies to breeding practice.
The present invention is to be achieved through the following technical solutions：
A method of building capsicum mutant library, including following operation using ethylmethane sulfonate：
1) full capsicum dry seeds are soaked seed 10~30min in 50~60 DEG C of warm water, is stirred continuously during seed soaking；
Pepper seed is taken out after seed soaking and blots surface moisture；
2) using the ethylmethane sulfonate solution that volume by volume concentration 0.6%~1.2%, pH are 7.0 as mutagenic treatment agent；It will be peppery
Green pepper seed is respectively placed in by grain in the mutagenic treatment agent of 0.5~0.8ml, and handling 8~16h in constant temperature oscillation under room temperature carries out
Mutagenic treatment；
3) pepper seed is taken out and is cleaned up after the completion of mutagenic treatment, proper alignment is in being decorated with grid of the same size
Moistening filter paper on, a seed, at room temperature light culture 14 days are placed in a grid；Simultaneously with the capsicum of non-mutagenic treatment
Seed equally processing is as a contrast；
It is not shorter than the half of seed length using the length of radicle as germination standard, counts the germinative number of pepper seed, and count
Calculate germination percentage and relative germination rate：
Germination percentage=(subnumber is planted experimentally in germinative number/confession) × 100%
Relative germination rate (%)=(germination percentage of germination percentage/control of mutagenic treatment) × 100%；
With relative germination rate 45~55% mutagenic treatment be most suitable mutagenic treatment；
4) nursery is carried out to the pepper seed for carrying out most suitable mutagenic treatment, Plant colonization is denoted as M in field after nursery1
Generation, normal field management divide single plant sowing, the seed of harvest to be denoted as M after seed maturity2Generation；
5)M2For seed and by strain sowing field planting, each strain is planted more plants, is throughout the growing season investigated often in plant
The leaf of a strain, stem, fruit, breeding time, floral organ, fertility phenotypic variation, to all phenotypic variation plant for screening by
The seed of single plant sowing, harvest is denoted as M3Generation；
6) each M3For seed by strain sowing field planting, each strain plants more plants, during plant strain growth, according to M2Generation
The characteristics of variant, is to M3Repeated measures and identification are carried out for the mutant character of strain；
7) M of heredity can be stablized to mutant character3Seed is harvested by single plant for strain, continues to plant by strain, it is ripe
The M for no longer detaching strain is harvested afterwards4For seed, all M for no longer detaching strain4Mutant library is constituted for seed.
The capsicum dry seeds are the dry seeds of capsicum annum fasciculatum.
The capsicum dry seeds are the dry seeds of " abiding by peppery No. 1 " capsicum annum fasciculatum.
The most suitable mutagenic treatment is：
Using the ethylmethane sulfonate solution that volume by volume concentration 0.8%, pH are 7.0 as mutagenic treatment agent；Pepper seed is pressed
Grain is respectively placed in the mutagenic treatment agent of 0.5~0.8ml, and mutagenic treatment is carried out in the processing of constant temperature oscillation under room temperature 12h.
The most suitable mutagenic treatment is：
Using the ethylmethane sulfonate solution that volume by volume concentration 1.0%, pH are 7.0 as mutagenic treatment agent；Pepper seed is pressed
Grain is respectively placed in the mutagenic treatment agent of 0.5~0.8ml, and mutagenic treatment is carried out in the processing of constant temperature oscillation under room temperature 10h；
The most suitable mutagenic treatment is：
Using the ethylmethane sulfonate solution that volume by volume concentration 1.2%, pH are 7.0 as mutagenic treatment agent；Pepper seed is pressed
Grain is respectively placed in the mutagenic treatment agent of 0.5~0.8ml, and mutagenic treatment is carried out in the processing of constant temperature oscillation under room temperature 8h.
Compared with prior art, the present invention has technique effect beneficial below：
The method provided by the invention for building capsicum mutant library using ethylmethane sulfonate is built using EMS mutagenesis and is surveyed
The mutant library of sequence kind such as (" abiding by peppery No. 1 ") can not only directly effectively utilize sequencing achievement and carry out Analysis of Mutants,
Clone for important agronomic trait gene creates conditions, and is established continuously to obtain the capsicum gene with independent intellectual property rights
Fixed basis, and obtain a plurality of types of basic materials；Pepper fruit can be promoted by having filtered out the mutant of fruit surface smooth
Marketing quality；Its fine hair layer of fine hair mutant has been filtered out as the natural cover for defense, aphid feeding and breeding can be hindered, promoted peppery
The aphis resistance energy of green pepper mitigates the virosis caused by aphid feeding and occurs；Having filtered out the compact mutant of plant type can improve
Plant type is conducive to rational close planting, realizes red pepper high-yield.
The selection of Induced dosage is the key that structure mutant library, and Induced dosage is the hair according to seed under various dose
What bud rate determined, therefore ensure that relatively accurate percentage of seedgermination is most important to structure mutant library.Influence percentage of seedgermination
Factor it is more, the either radioinduction of capsicum or EMS mutagenesises in the prior art, without excluding space to hair
The influence of bud rate.When determining Induced dosage, mutation time is another vital influence factor.The present invention uses
EMS carries out mutagenesis to capsicum, and the treatment fluid dosage of every seed, explores not when specifying EMS mutagenic treatment capsicum annum fasciculatums
Influence of the same processing time to germination percentage after mutagenesis, while eliminating influence of the space to germination percentage.
With on current capsicum frequently with radioinduction compared with, EMS of the invention generate mutation based on point mutation, it is right
Genomic damage is small, lethal mutation is few, the spectrum of mutation is stable, the frequency of mutation is high, and without expensive instrument and equipment, and dosage is easy
Control, operation is simple, and cost is relatively low.
There is researcher to point out that the half lethal dose of capsicum is the EMS solution treatment 4h of mass fraction 1.0%, constructs capsicum
6421 mutant library obtains leaf, stalk, fruit, fertility, 5 class mutant of breeding time (week book etc., 2017), discloses
EMS handles the validity of capsicum.In contrast, capsicum 6421 is long red pepper, and genome composition is different from capsicum annum fasciculatum, because of EMS
It is to generate phenotypic variation by changing genome sequence so that the EMS half lethal doses and Mutagenic Effect of it and capsicum annum fasciculatum are completely
It is different.And in EMS mutagenic treatment capsicums the treatment fluid dosage of every seed and different processing times to mutagenesis after germination percentage
Influence it is still unknown in the case of, structure capsicum mutant library may effect it is not good enough.The present invention by grain to pepper seed into
For row EMS treatment fluids to carry out mutagenesis, wherein EMS treatment fluids ensure enough abundant immersions, then utilize the picture size one on filter paper
The grid of cause excludes influence of the space to germination percentage, peppery under different mutation times by comparing various concentration EMS treatment fluids
Green pepper percentage of seedgermination, determines half lethal dose, half lethal dose mutagenic treatment pepper seed is used in combination, in M2The generation investigation frequency of mutation
And mutation type, M3、M4Repeated measures and identification phenotypic variation are obtained with the mutation construction mutant library for stablizing heredity
The types mutant such as leaf, stem, fruit, breeding time, floral organ, fertility formulates abundant material for capsicum functional genomics research
Material, while fractional mutant also may be directly applied to breeding practice.
Description of the drawings
Fig. 1 is the schematic diagram that EMS carries out capsicum mutagenesis on the filter paper for being decorated with grid in the same size；
Fig. 2 is the capsicum fruit surface smooth type mutant of EMS inductions；The left side is the mutant of fruit surface smooth in figure, and the right is
Control；
Fig. 3 is the capsicum fine hair type mutant of EMS inductions；It is above control in figure, is fine hair mutant below；
Fig. 4 is the capsicum plant type compact mutant of EMS inductions；The left side is control in figure, and the right is that plant type becomes compact
Mutant.
Specific implementation mode
With reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.
1, the preparation of EMS treatment fluids：With 305ml, the disodium phosphate soln of a concentration of 0.2mol/L and 195ml, concentration
For the sodium dihydrogen phosphate mixing of 0.2mol/L, it is separately added into the EMS of 0,6,7,8,9,10,11 and 12ml, it is fixed with distilled water
Hold to 1 liter, volume by volume concentration be 0,0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, pH is 7.0
EMS mutagenic treatment liquid.
2, EMS mutagenic treatments：Full " abiding by peppery No. 1 " capsicum annum fasciculatum is chosen (to be carried by Zunyi academy of agricultural sciences Yu Changshui researcher
For) dry seeds blot the moisture of the surface of the seed with filter paper with 50~60 DEG C of water seed soaking 15min, it is 0 with volume by volume concentration,
The EMS treatment fluids that 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, pH are 7.0 distinguish mutagenic treatment 8h,
10h, 12h, 14h, 16h, keep 1 0.5~0.8ml of seed treatment fluid (key of Induced dosage be concentration for the treatment of and when
Between, treatment fluid ensures that seed is fully soaked seed), entire mutagenic processes are placed in 26 DEG C, in the shaking table of 110rpm at constant temperature oscillation
Reason, then takes out seed, is rinsed 5 times with pure water, becomes the pepper seed after EMS mutagenic treatments.
3, the germination test of seed：
The seed of above-mentioned EMS mutagenic treatments is placed in the culture dish (a diameter of 90mm) for 2 layers of filter paper for being covered with wetting,
In above one layer of filter paper be decorated with 50 grids of the same size, as shown in Figure 1, carrying out germination test, each processing sets 3 weights
It is multiple, 50 seeds are each repeated, 26 DEG C of light cultures in growth cabinet are placed in, are not shorter than the one of seed length with the length of radicle
Half is germination standard, germination percentage and relative germination rate when recording chitting piece number daily, and calculating 14 days, by relative germination rate
Treatment dosage close to 50% is determined as optimum Induced dosage, i.e. half lethal dose.
The computational methods of germination percentage and relative germination rate are as follows：
Germination percentage (%)=(all subnumber is planted experimentally in normal germinative number/confession) × 100%；
Germination percentage × 100% of germination percentage/control of relative germination rate (%)=processing；
It is as shown in table 1 to count the pepper seed germination rate obtained under different EMS concentration and mutation time.
Pepper seed germination rate under table 1 difference EMS concentration and mutation time
Select following three processing as optimum Induced dosage：0.8% processing 12h, 1.0% processing 10h, 1.2%
Handle 8h.
4, aberration rate and distortion type investigation：
One of half lethal dose (0.8%, 12h) is selected to carry out EMS mutagenic treatments to pepper seed, through mutagenic treatment
Pepper seed afterwards, Plant colonization is denoted as M in field after nursery1Generation, normal field management divide single plant sowing after seed maturity,
The seed of harvest is denoted as M2Generation, each M2In generation, by strain sowing field planting, the phenotype of each strain was throughout the growing season investigated in plant
Variation, to the phenotypic variation plant that screens by single plant sowing, the seed of harvest is denoted as M3Generation, each M3In generation, is fixed by strain sowing
It plants, each strain plants more plants, during plant strain growth, according to M2The characteristics of for variant, is to M3It is carried out for the stability of strain
Identification, the single plant that can stablize heredity to mutant character continue to plant M4, strain seed is harvested after ripe, constitutes mutant library.
Specific embodiment is given below
Embodiment 1
(1) preparation of EMS treatment fluids：With 305ml, the disodium phosphate soln of a concentration of 0.2mol/L and 195ml, concentration
For the sodium dihydrogen phosphate mixing of 0.2mol/L, it is separately added into the EMS of 0,6,7,8,9,10,11 and 12ml, it is fixed with distilled water
Hold to 1 liter, volume by volume concentration be 0,0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, pH is 7.0
EMS mutagenic treatment liquid.
(2) EMS mutagenic treatments：
A. full " abiding by peppery No. 1 " capsicum annum fasciculatum dry seeds are chosen, every 50 are 1 part, are respectively charged into the nylon wire of 15*10cm
Bag, is put into 55 DEG C of warm water and is stirred continuously, and soak seed 15min；
B. the surface moisture of every part of seed after hoting water treatment of seeds is blotted with filter paper；
C. it is 0%, 0.6%, 0.7%, 0.8%, 0.9% that volume by volume concentration is injected separately into 50ml round bottom centrifuge tubes,
The EMS mutagenic treatment liquid that 1.0%, 1.1%, 1.2%, pH are 7.0, the seed that surface moisture is blotted is placed in one, and keeps 1
The treatment fluid of grain seed 0.6ml is individually positioned in 26 DEG C, and constant temperature oscillation handles 8h, 10h, 12h, 14h in the shaking table of 110rpm,
16h, totally 5 controls and 35 processing, each control and processing are 50 seeds, and are 3 repetitions；
D. EMS mutagenic treatment liquid is outwelled, seed pure water is rinsed 5 times, remaining EMS is removed, becomes through EMS mutagenesis
Treated pepper seed.
(3) germination test of seed：
A. 50 grids of the same size are drawn on the circular filter paper of a diameter of 90mm, as shown in Figure 1, by 2 filter paper
Overlapping has one of grid upper, is put into the culture dish of a diameter of 90mm, suitable pure water is added and ensures filter paper moistening；
B. by grid of the pepper seed proper alignment on filter paper after EMS mutagenic treatments, 50 seeds correspond to 50
A grid is placed in 26 DEG C of light cultures in growth cabinet；
C. chitting piece number is recorded daily, and after 14 days, the half for not being shorter than seed length with the length of radicle is that germination is marked
Standard, counts the germinative number of 5 controls and 35 processing, and calculates germination percentage and relative germination rate：
Germination percentage (%)=(subnumber is planted experimentally in normal germinative number/confession when 14D) × 100%
Relative germination rate (%)=(germination percentage of germination percentage/control of processing) × 100%；
D. relative germination rate is determined as optimum Induced dosage, i.e. half lethal dose close to 50% treatment dosage：
0.8% processing 12h, 1.0% processing 10h, 1.2% processing 8h.
(4) aberration rate and distortion type investigation：
A. in the method for step (2) with half lethal dose (0.8%, 12h) to 5250 " abiding by peppery No. 1 " capsicum annum fasciculatum dry seeds
Carry out mutagenic treatment；
B. the pepper seed after EMS mutagenic treatments, Plant colonization is denoted as M in field after nursery1Generation, normal field pipe
It manages, divides single plant sowing after seed maturity, harvest the seed of 2271 single plants altogether, the seed of harvest is denoted as M2Generation；
C. 295 M are selected at random2In generation, each strain planted 12 plants, in plant entire growth period by strain sowing field planting
Between, " abiding by peppery No. 1 " not carry out mutagenic treatment is control, investigate the leaf of each strain, stem, fruit, breeding time, floral organ,
The phenotypic variation of fertility shares 94 strains and performance variation occurs, and the whole frequency of mutation is 31.86%, the middle period, stem, fruit,
Breeding time, floral organ, fertility account for respectively the whole frequency of mutation 20%, 34.55%, 21.82%, 12.12%, 4.24%,
7.27%, to the phenotypic variation plant that screens by single plant sowing, the seed of harvest is denoted as M3Generation；
D. each M3In generation, each strain planted 12 plants, during plant strain growth, according to M by strain sowing field planting2Generation variation
The characteristics of strain, is to M3Repeated measures and identification are carried out for the mutant character of strain；
F. the single plant that can stablize heredity to mutant character continues to plant M4, the kind for no longer detaching strain is harvested after ripe
The seed of son, all no longer separation strains constitutes mutant library.
Present invention obtains the types mutant such as leaf, stem, fruit, breeding time, floral organ, fertility, are capsicum functional gene
Group learns research and formulates abundant material, while fractional mutant also may be directly applied to breeding practice, such as in M2In generation, occurs as soon as
The elongated mutant of the mutant of fruit surface smooth, fine hair, plant type compact mutant, single plant is reserved seed for planting, the seed note of harvest
For M3Generation, M3In generation, is colonized by strain, and each strain plants 12 plants, M3It is no longer detached for character, single plant is reserved seed for planting, the seed note of harvest
For M4Generation, M4Generation mixing sowing, institute sowing corresponding mutant seeds.
Wherein, the phenotype of the mutant of fruit surface smooth and the fruit before mutation face than as shown in Fig. 2, the left side is fruit face in figure
Smooth mutant, the right are control；It can be seen that the fruit type in mutant remains unchanged but relatively smooth, the fruit of fruit face variation
Fold on face significantly reduces, and the reflective degree in fruit face is also remarkably reinforced, and the mutant of fruit surface smooth improves the quotient of pepper fruit
Product quality, it will product is contributed to occupy the market of bigger.
The phenotype of fine hair mutant, as shown in figure 3, above to compare in figure, is fine hair below with the limb comparison before mutation
Mutant；It can be seen that intensive fine hair layer occurs in the limb surface layer of fine hair mutant, and fine hair layer is as the natural cover for defense, can
Aphid feeding and breeding are hindered, the aphis resistance energy of capsicum is promoted, mitigates the virosis caused by aphid feeding and occurs；Also corresponding
The administration number of times for reducing pesticide, dosage.
With the plant type comparison before mutation as shown in figure 4, the left side is to compare in figure, the right is the compact mutation type surface of plant type
Plant type becomes compact mutant.It can be seen that the level inclination of the branches and tendrils of the compact mutant of plant type significantly increases, between limb
Spacing is reduced, and is conducive to rational close planting, and the compact mutant of plant type can improve plant type, realize red pepper high-yield.
Example given above is to realize the present invention preferably example, and the present invention is not limited to the above embodiments.This field
Technical staff any nonessential addition, the replacement made according to the technical characteristic of technical solution of the present invention, belong to this
The protection domain of invention.
Claims (6)
1. a kind of method building capsicum mutant library using ethylmethane sulfonate, which is characterized in that including following operation：
1) full capsicum dry seeds are soaked seed 10~30min in 50~60 DEG C of warm water, is stirred continuously during seed soaking；Seed soaking
After take out and pepper seed and blot surface moisture；
2) using the ethylmethane sulfonate solution that volume by volume concentration 0.6%~1.2%, pH are 7.0 as mutagenic treatment agent；By capsicum kind
Son is respectively placed in by grain in the mutagenic treatment agent of 0.5~0.8ml, and handling 8~16h in constant temperature oscillation under room temperature carries out mutagenesis
Processing；
3) pepper seed is taken out and is cleaned up after the completion of mutagenic treatment, proper alignment is in being decorated with the wet of grid of the same size
Moisten on filter paper, a seed is placed in a grid, at room temperature light culture 14 days；Simultaneously with the pepper seed of non-mutagenic treatment
Same processing is as a contrast；
It is not shorter than the half of seed length using the length of radicle as germination standard, counts the germinative number of pepper seed, and calculate hair
Bud rate and relative germination rate：
Germination percentage=(subnumber is planted experimentally in germinative number/confession) × 100%；
Relative germination rate (%)=(germination percentage of germination percentage/control of mutagenic treatment) × 100%；
With relative germination rate 45~55% mutagenic treatment be most suitable mutagenic treatment；
4) nursery is carried out to the pepper seed for carrying out most suitable mutagenic treatment, Plant colonization is denoted as M in field after nursery1Generation, just
Normal field management divides single plant sowing, the seed of harvest to be denoted as M after seed maturity2Generation；
5) M is selected2For seed and by strain sowing field planting, each strain is planted more plants, is throughout the growing season investigated each in plant
The leaf of strain, stem, fruit, breeding time, floral organ, fertility phenotypic variation, to all phenotypic variation plant for screening by list
Strain sowing, the seed of harvest are denoted as M3Generation；
6) each M3For seed by strain sowing field planting, each strain plants more plants, during plant strain growth, according to M2Generation variation
The characteristics of strain, is to M3Repeated measures and identification are carried out for the mutant character of strain；
7) M of heredity can be stablized to mutant character3Seed is harvested by single plant for strain, continues to plant by strain, be harvested after ripe
No longer detach the M of strain4For seed, all M for no longer detaching strain4Mutant library is constituted for seed.
2. utilizing the method for ethylmethane sulfonate structure capsicum mutant library as described in claim 1, which is characterized in that described
Capsicum dry seeds be capsicum annum fasciculatum dry seeds.
3. utilizing the method for ethylmethane sulfonate structure capsicum mutant library as claimed in claim 1 or 2, which is characterized in that
The capsicum dry seeds are the dry seeds of " abiding by peppery No. 1 " capsicum annum fasciculatum.
4. utilizing the method for ethylmethane sulfonate structure capsicum mutant library as described in claim 1, which is characterized in that described
Most suitable mutagenic treatment be：
Using the ethylmethane sulfonate solution that volume by volume concentration 0.8%, pH are 7.0 as mutagenic treatment agent；By pepper seed by grain point
It is not placed in the mutagenic treatment agent of 0.5~0.8ml, mutagenic treatment is carried out in the processing of constant temperature oscillation under room temperature 12h.
5. utilizing the method for ethylmethane sulfonate structure capsicum mutant library as described in claim 1, which is characterized in that described
Most suitable mutagenic treatment be：
Using the ethylmethane sulfonate solution that volume by volume concentration 1.0%, pH are 7.0 as mutagenic treatment agent；By pepper seed by grain point
It is not placed in the mutagenic treatment agent of 0.5~0.8ml, mutagenic treatment is carried out in the processing of constant temperature oscillation under room temperature 10h.
6. utilizing the method for ethylmethane sulfonate structure capsicum mutant library as described in claim 1, which is characterized in that described
Most suitable mutagenic treatment be：
Using the ethylmethane sulfonate solution that volume by volume concentration 1.2%, pH are 7.0 as mutagenic treatment agent；By pepper seed by grain point
It is not placed in the mutagenic treatment agent of 0.5~0.8ml, mutagenic treatment is carried out in the processing of constant temperature oscillation under room temperature 8h.
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| CN110100728A (en) * | 2019-05-21 | 2019-08-09 | 中国农业科学院烟草研究所 | A method of mutant library is generated by EMS mutagenesis bluish dogbane |
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| CN112790106A (en) * | 2019-11-13 | 2021-05-14 | 江苏沿海地区农业科学研究所 | Method for constructing barley mutant library by using Ethyl Methane Sulfonate (EMS) |
| CN113349052A (en) * | 2021-07-12 | 2021-09-07 | 中国海洋大学 | Method for constructing laver mutant library |
| CN115777530A (en) * | 2022-12-20 | 2023-03-14 | 黑龙江省原子能研究院 | Screening method for semi-lethal dose of dormant seeds of physical compound mutagenesis straight root system crops |
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| CN112205294A (en) * | 2019-11-12 | 2021-01-12 | 广东石油化工学院 | Mutation breeding method for cyperus esculentus |
| CN112790106A (en) * | 2019-11-13 | 2021-05-14 | 江苏沿海地区农业科学研究所 | Method for constructing barley mutant library by using Ethyl Methane Sulfonate (EMS) |
| CN113349052A (en) * | 2021-07-12 | 2021-09-07 | 中国海洋大学 | Method for constructing laver mutant library |
| CN115777530A (en) * | 2022-12-20 | 2023-03-14 | 黑龙江省原子能研究院 | Screening method for semi-lethal dose of dormant seeds of physical compound mutagenesis straight root system crops |
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