CN112470932A - Method for breeding new salt-tolerant lily strain by using EMS reagent mutagenesis - Google Patents
Method for breeding new salt-tolerant lily strain by using EMS reagent mutagenesis Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The invention discloses a method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis, which comprises the following steps: a. inducing lily embryonic callus; b. preparing a salt-tolerant mutagenesis culture medium; c. salt-tolerant mutagenesis: putting the callus on a salt-tolerant mutagenesis culture medium containing an EMS reagent for mutagenesis; d. inducing to form seedlings: sequentially inoculating the surviving callus onto a seedling culture medium and a bulb culture medium for culture, and growing lily bulblet with the diameter of 0.1cm-0.5cm for later use; e. salt-tolerant screening: transferring the cultured lily bulblet with the diameter of 0.1cm-0.5cm to a screening culture medium for culture, wherein the salt-tolerant lily bulb growing normally is the new salt-tolerant lily strain. The method for breeding the new salt-tolerant lily strain by performing the directional mutagenesis under the salt environment culture condition has the advantages of simple operation, strong operability and high mutagenesis utilization efficiency, and can greatly enrich lily salt-tolerant germplasm resources in a short time.
Description
Technical Field
The invention relates to the technical field of seedlings, in particular to a method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis, which can efficiently culture a new lily salt-tolerant germplasm.
Background
With the expansion of the world lily production and seedball trade, the lily production area and seedball demand, however, the lily growth conditions are strict, and the lily is favored to be suitable for growing in a slightly acidic environment with good drainage and is sensitive to salt. The salinity of the soil is a key factor related to the normal growth of the lilies, and the water and soil in most northern areas of China are alkaline with partial salt, so that the development of the lilies industry is greatly limited.
At present, the research on lily salt-tolerant varieties mainly focuses on the evaluation of a salt-tolerant system and field screening, but has the problems of less germplasm resources, slow progress of salt-tolerant strains and the like, and the development requirement of the lily industry is difficult to meet.
Disclosure of Invention
The invention aims to provide a method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis, aiming at the defects in the prior art, and the method can be used for efficiently breeding new salt-tolerant lily germplasm.
The invention aims to solve the problems by the following technical scheme:
a method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis is characterized by comprising the following steps: the method comprises the following steps:
a. induction of lily embryonic callus: placing the lily scales on an induction callus culture medium to induce faint yellow callus particles to appear for later use;
b. preparing a salt-tolerant mutagenesis culture medium: preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation;
c. salt-tolerant mutagenesis of callus: c, placing the callus prepared in the step a on the salt-tolerant mutagenesis culture medium containing the EMS reagent prepared in the step b, and carrying out salt-tolerant mutagenesis;
d. inducing to form seedlings: inoculating the callus survived by salt-tolerant mutagenesis to a seedling culture medium for culture, transferring lily bulblets growing on the callus to a bulb culture medium for continuous culture, and reserving when lily bulblets with the diameter of 0.1cm-0.5cm grow for later use;
e. salt-tolerant screening: transferring the cultured lily bulblet with the diameter of 0.1cm-0.5cm to a screening culture medium for salt-tolerant screening culture, wherein the formula of the screening culture medium is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60g/L, and after 35-40 days, screening out the salt-tolerant lily bulb which normally grows, namely the new salt-tolerant lily strain.
The lily scales in the step a are processed as follows: washing the lily scales with running water for 0.5-2 hours; soaking in 75% alcohol for 30-40s, adding into 0.1-0.2% mercuric chloride for 7-9min, washing with sterile water for 4-6 times, cutting the scale into small pieces with concave surface facing upwards, and placing on callus induction culture medium for induction culture.
The size of the small pieces was 0.5cm by 0.5 cm.
The culture conditions of the lily scales in the step a are as follows: dark culture at 23-26 deg.C for 50-60 days.
The preparation method of the salt-tolerant mutagenesis culture medium in the step b comprises the following steps: firstly, preparing a mutation culture medium of MS +5mg/L2,4-D +2g/L NaCL +6.5g/L agar and 30g/L sucrose, wherein the pH of the mutation culture medium is = 5.8-6.0; after the mutagenesis culture medium is sterilized at high temperature, the mutagenesis culture medium is kept at the temperature of more than 50 ℃ and is in a liquid state; adding EMS mother liquor into a sterile fume hood, preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation.
The process of adding the EMS mother liquor requires wearing a radiation protection work clothes and wearing a gas mask, and adopts a disposable filter with the diameter of 0.22 mu m to add the EMS mother liquor.
The callus in the step c is cut into soybean (0.2cm x 0.2cm) under aseptic condition, and then placed on a salt-tolerant mutagenesis culture medium containing EMS reagent for salt-tolerant mutagenesis, wherein the culture condition is light and dark culture time of 10h/14h and culture temperature of 23-26 ℃.
The formula of the seedling culture medium in the step d is 1/2MS +0.2mg/L6-BA +0.1mg/LNAA +6.5g/L agar +15g/L sucrose, and the pH of the seedling culture medium is = 5.8-6.0; the formula of the bulb culture medium in the step d is as follows: MS +0.1mg/LNAA +6.5g/L agar +60g/L sucrose.
The formula of the screening culture medium in the step e is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60 g/L.
The process of carrying out propagation culture on the salt-tolerant lily new strain in the step e comprises the following steps: carrying out plant propagation on the screened salt-tolerant lily bulbs by plant, and after the propagation reaches a certain amount, removing dormancy at low temperature to plant and observe the bulbs in the field; the propagation culture medium comprises: MS +0.1mg/LNAA +1mg/L6-BA +6.5g/L agar +60g/L sucrose, pH of the propagation medium = 5.8-6.0.
Compared with the prior art, the invention has the following advantages:
the invention provides a method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis, which aims to overcome the difficulty in breeding the existing lily salt-tolerant variety, and provides a brand-new method for breeding the new salt-tolerant lily strain by performing directional mutagenesis under the salt environment culture condition.
Detailed Description
The present invention will be further described with reference to the following examples.
A method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis comprises the following steps:
a. induction of lily embryonic callus
Washing the lily scales with running water for 0.5-2 hours; soaking in 75% alcohol for 30-40s, adding into 0.1-0.2% mercuric chloride for 7-9min, washing with sterile water for 4-6 times, cutting the scale into 0.5cm x 0.5cm pieces, and culturing on inducing callus culture medium with concave surface facing upwards; culturing in dark at 23-26 deg.C for 50-60d to induce faint yellow callus particles; the callus induction culture medium is as follows: MS +5mg/L2,4-D +6.5g/L agar +30g/L sucrose, pH of the medium = 5.8-6.0;
b. preparing salt-tolerant mutagenesis culture medium
Preparing a mutation culture medium of MS +5mg/L2,4-D +2g/L NaCL +6.5g/L agar and 30g/L sucrose in advance, wherein the pH of the mutation culture medium is = 5.8-6.0; after the mutagenesis culture medium is sterilized at high temperature, the mutagenesis culture medium is kept at the temperature of more than 50 ℃ and is in a liquid state; putting on a radiation protection working clothes and taking a gas mask, adding EMS (ethyl methyl xanthate) mother liquor by using a disposable filter (the diameter is 0.22 mu m) in an aseptic fume hood, and preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation;
c. callus salt tolerance mutagenesis
B, cutting the callus prepared in the step a into soybean (0.2cm x 0.2cm) under an aseptic condition, placing the soybean on a salt-tolerant mutagenesis culture medium containing an EMS reagent for salt-tolerant mutagenesis, wherein the culture condition is that the light-dark culture time is 10h/14h, the culture temperature is 23-26 ℃, and observing the survival rate of the callus in the salt-tolerant mutagenesis culture medium after culturing for at least 40 days, and the survival rate is generally 25-35%;
d. inducing seedling
Inoculating the salt-tolerant mutagenized and surviving callus onto a seedling culture medium, wherein the formula of the seedling culture medium is 1/2MS +0.2mg/L6-BA +0.1mg/LNAA +6.5g/L agar +15g/L sucrose, and the pH of the seedling culture medium is = 5.8-6.0; after 40 days, transferring the lily bulblets growing on the callus onto a bulb culture medium, wherein the formula of the bulb culture medium is as follows: MS +0.1mg/LNAA +6.5g/L agar +60g/L sucrose, and growing lily bulblet with diameter of 0.1cm-0.5cm for standby after 50-55 days;
e. salt tolerant screening
Transferring the cultured lily bulblet with the diameter of 0.1cm-0.5cm to a screening culture medium for salt-tolerant screening culture, wherein the formula of the screening culture medium is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60g/L, after 35-40 days, screening out salt-tolerant lily bulbs which can normally grow, namely the new salt-tolerant lily strain;
f. propagation of new salt-tolerant lily strain
Carrying out plant propagation on the screened salt-tolerant lily bulbs by plant, and after the propagation reaches a certain amount, removing dormancy at low temperature to plant and observe the bulbs in the field; the propagation culture medium comprises: MS +0.1mg/LNAA +1mg/L6-BA +6.5g/L agar +60g/L sucrose, pH of the propagation medium = 5.8-6.0.
The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis provided by the invention is further described by the following specific embodiment.
Example A variety "Sobang (sorbone)"
The method for breeding the new strain of the salt-tolerant Lily Sophora by using EMS reagent mutagenesis comprises the following steps:
a. induction of lily embryonic callus
Washing the scale of the Sobang lily with running water for 1.5 hours; soaking in 75% ethanol for 30-40s, adding into 0.2% mercuric chloride for 7-9min, washing with sterile water for 4-6 times, cutting the scale into 0.5cm by 0.5cm pieces, and culturing on inducing callus culture medium with concave surface facing upwards; culturing in dark at 23-26 deg.C for 50-60d to induce faint yellow callus particles; the callus induction culture medium is as follows: MS +5mg/L2,4-D +6.5g/L agar +30g/L sucrose, pH of the medium = 5.8;
b. preparing salt-tolerant mutagenesis culture medium
A mutagenesis medium of MS +5mg/L2,4-D +2g/L NaCL +6.5g/L agar +30g/L sucrose is prepared in advance, and the pH of the mutagenesis medium is = 5.8; after the mutagenesis culture medium is sterilized at high temperature, the mutagenesis culture medium is kept at the temperature of more than 50 ℃ and is in a liquid state; putting on a radiation protection working clothes and taking a gas mask, adding EMS (ethyl methyl xanthate) mother liquor by using a disposable filter (the diameter is 0.22 mu m) in an aseptic fume hood, and preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation;
c. callus salt tolerance mutagenesis
B, cutting the callus prepared in the step a into 0.2cm x 0.2cm under aseptic conditions, placing the callus on a salt-tolerant mutagenesis culture medium containing an EMS reagent for salt-tolerant mutagenesis, wherein the culture conditions comprise light and dark culture time of 10h/14h and culture temperature of 23-26 ℃, and observing the survival rate of the callus in the salt-tolerant mutagenesis culture medium to be 34% after culturing for 45 d;
d. inducing seedling
Inoculating the callus survived by salt-tolerant mutagenesis on a seedling culture medium, wherein the formula of the seedling culture medium is 1/2MS +0.2mg/L6-BA +0.1mg/LNAA +6.5g/L agar +15g/L sucrose, the pH of the seedling culture medium is =5.8, after 40 days, transferring lily bulblet growing on the callus onto a bulb culture medium, and the formula of the bulb culture medium is as follows: MS +0.1mg/LNAA +6.5g/L agar +60g/L sucrose, and growing lily bulblet with diameter of 0.1cm-0.5cm for standby after 55 days;
e. salt tolerant screening
Transferring the cultured lily bulblet with the diameter of 0.1cm-0.5cm to a screening culture medium for salt-tolerant screening culture, wherein the formula of the screening culture medium is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60g/L, 35d later, screening out salt-tolerant lily bulbs which can normally grow, namely the new salt-tolerant lily strain;
f. propagation of new salt-tolerant lily strain
Carrying out plant propagation on the screened salt-tolerant lily bulbs by plant, and after the propagation reaches a certain amount, removing dormancy at low temperature to plant and observe the bulbs in the field; the propagation culture medium comprises: MS +0.1mg/LNAA +1mg/L6-BA +6.5g/L agar +60g/L sucrose, pH of the propagation medium = 5.8.
EXAMPLE two varieties of wooden door (concad' or) "
The method for breeding the new variety of the salt-tolerant wooden door lily by using EMS reagent mutagenesis comprises the following steps:
a. induction of lily embryonic callus
Washing the lily scales of the wooden door with running water for 0.5-2 hours; soaking in 75% alcohol for 30-40s, adding into 0.1-0.2% mercuric chloride for 7-9min, washing with sterile water for 4-6 times, cutting the scale into 0.5cm x 0.5cm pieces, and culturing on inducing callus culture medium with concave surface facing upwards; culturing in dark at 23-26 deg.C for 50-60d to induce faint yellow callus particles; the callus induction culture medium is as follows: MS +5mg/L2,4-D +6.5g/L agar +30g/L sucrose, pH of the medium = 5.9;
b. preparing salt-tolerant mutagenesis culture medium
A mutagenesis medium of MS +5mg/L2,4-D +2g/L NaCL +6.5g/L agar +30g/L sucrose is prepared in advance, and the pH of the mutagenesis medium is = 5.9; after the mutagenesis culture medium is sterilized at high temperature, the mutagenesis culture medium is kept at the temperature of more than 50 ℃ and is in a liquid state; putting on a radiation protection working clothes and taking a gas mask, adding EMS (ethyl methyl xanthate) mother liquor by using a disposable filter (the diameter is 0.22 mu m) in an aseptic fume hood, and preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation;
c. callus salt tolerance mutagenesis
B, cutting the callus prepared in the step a into 0.2cm x 0.2cm under aseptic conditions, placing the callus on a salt-tolerant mutagenesis culture medium containing an EMS reagent for salt-tolerant mutagenesis, wherein the culture conditions comprise light and dark culture time of 10h/14h and culture temperature of 23-26 ℃, and observing the survival rate of the callus in the salt-tolerant mutagenesis culture medium to be 31% after culturing for 40 d;
d. inducing seedling
Inoculating the callus survived by salt-tolerant mutagenesis on a seedling culture medium, wherein the formula of the seedling culture medium is 1/2MS +0.2mg/L6-BA +0.1mg/LNAA +6.5g/L agar +15g/L sucrose, the pH of the seedling culture medium is =5.9, after 40 days, transferring lily bulblet growing on the callus onto a bulb culture medium, and the formula of the bulb culture medium is as follows: MS +0.1mg/LNAA +6.5g/L agar +60g/L sucrose, and growing lily bulblet with diameter of 0.1cm-0.5cm for standby after 50 days;
e. salt tolerant screening
Transferring the cultured lily bulblet with the diameter of 0.1cm-0.5cm to a screening culture medium for salt-tolerant screening culture, wherein the formula of the screening culture medium is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60g/L, and after 38d, screening out salt-tolerant lily bulbs capable of growing normally, namely the new salt-tolerant lily strain;
f. propagation of new salt-tolerant lily strain
Carrying out plant propagation on the screened salt-tolerant lily bulbs by plant, and after the propagation reaches a certain amount, removing dormancy at low temperature to plant and observe the bulbs in the field; the propagation culture medium comprises: MS +0.1mg/LNAA +1mg/L6-BA +6.5g/L agar +60g/L sucrose, pH of the propagation medium = 5.9.
The invention provides a method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis, which aims to overcome the difficulty in breeding the existing lily salt-tolerant variety, and provides a brand-new method for breeding the new salt-tolerant lily strain by performing directional mutagenesis under the salt environment culture condition.
The above embodiments are only for illustrating the technical idea of the present invention, and the protection scope of the present invention cannot be limited thereby, and any modification made on the basis of the technical scheme according to the technical idea proposed by the present invention falls within the protection scope of the present invention; the technology not related to the invention can be realized by the prior art.
Claims (10)
1. A method for breeding a new salt-tolerant lily strain by using EMS reagent mutagenesis is characterized by comprising the following steps: the method comprises the following steps:
a. induction of lily embryonic callus: placing the lily scales on an induction callus culture medium to induce faint yellow callus particles to appear for later use;
b. preparing a salt-tolerant mutagenesis culture medium: preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation;
c. salt-tolerant mutagenesis of callus: c, placing the callus prepared in the step a on the salt-tolerant mutagenesis culture medium containing the EMS reagent prepared in the step b, and carrying out salt-tolerant mutagenesis;
d. inducing to form seedlings: inoculating the callus survived by salt-tolerant mutagenesis to a seedling culture medium for culture, transferring lily bulblets growing on the callus to a bulb culture medium for continuous culture, and reserving when lily bulblets with the diameter of 0.1cm-0.5cm grow for later use;
e. salt-tolerant screening: transferring the cultured lily bulblet with the diameter of 0.1cm-0.5cm to a screening culture medium for salt-tolerant screening culture, wherein the formula of the screening culture medium is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60g/L, and after 35-40 days, screening out the salt-tolerant lily bulb which normally grows, namely the new salt-tolerant lily strain.
2. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the lily scales in the step a are processed as follows: washing the lily scales with running water for 0.5-2 hours; soaking in 75% alcohol for 30-40s, adding into 0.1-0.2% mercuric chloride for 7-9min, washing with sterile water for 4-6 times, cutting the scale into small pieces with concave surface facing upwards, and placing on callus induction culture medium for induction culture.
3. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 2, which is characterized in that: the size of the small pieces was 0.5cm by 0.5 cm.
4. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the culture conditions of the lily scales in the step a are as follows: dark culture at 23-26 deg.C for 50-60 days.
5. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the preparation method of the salt-tolerant mutagenesis culture medium in the step b comprises the following steps: firstly, preparing a mutation culture medium of MS +5mg/L2,4-D +2g/L NaCL +6.5g/L agar and 30g/L sucrose, wherein the pH of the mutation culture medium is = 5.8-6.0; after the mutagenesis culture medium is sterilized at high temperature, the mutagenesis culture medium is kept at the temperature of more than 50 ℃ and is in a liquid state; adding EMS mother liquor into a sterile fume hood, preparing a salt-tolerant mutagenesis culture medium: MS +5mg/L2,4-D +2g/L NaCl +6.5g/L agar +30g/L sucrose +0.5% EMS, and subpackaging for later use after preparation.
6. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 5, which is characterized in that: the process of adding the EMS mother liquor requires wearing a radiation protection work clothes and wearing a gas mask, and adopts a disposable filter with the diameter of 0.22 mu m to add the EMS mother liquor.
7. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the callus in the step c is cut into the size of soybeans under the aseptic condition, and then placed on a salt-tolerant mutagenesis culture medium containing an EMS reagent for salt-tolerant mutagenesis, wherein the culture condition is that the light-dark culture time is 10h/14h, and the culture temperature is 23-26 ℃.
8. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the formula of the seedling culture medium in the step d is 1/2MS +0.2mg/L6-BA +0.1mg/LNAA +6.5g/L agar +15g/L sucrose, and the pH of the seedling culture medium is = 5.8-6.0; the formula of the bulb culture medium in the step d is as follows: MS +0.1mg/LNAA +6.5g/L agar +60g/L sucrose.
9. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the formula of the screening culture medium in the step e is as follows: MS +0.1mg/LNAA +4g/LNaCL +6.5g/L agar +60 g/L.
10. The method for breeding the new salt-tolerant lily strain by using EMS reagent mutagenesis as claimed in claim 1, which is characterized in that: the process of carrying out propagation culture on the salt-tolerant lily new strain in the step e comprises the following steps: carrying out plant propagation on the screened salt-tolerant lily bulbs by plant, and after the propagation reaches a certain amount, removing dormancy at low temperature to plant and observe the bulbs in the field; the propagation culture medium comprises: MS +0.1mg/LNAA +1mg/L6-BA +6.5g/L agar +60g/L sucrose, pH of the propagation medium = 5.8-6.0.
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Application publication date: 20210312 |