CN106417000B - A kind of breeding method of no pollen lily cultivar - Google Patents
A kind of breeding method of no pollen lily cultivar Download PDFInfo
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- CN106417000B CN106417000B CN201610852456.XA CN201610852456A CN106417000B CN 106417000 B CN106417000 B CN 106417000B CN 201610852456 A CN201610852456 A CN 201610852456A CN 106417000 B CN106417000 B CN 106417000B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The present invention relates to a kind of breeding methods of no pollen lily cultivar, belong to the new variety selective breeding technology field of plant.Lily is the main commercial floriculture in the world, in plantation extensively all over China, occupies the huge market share.But since lilium pollen is easier to pollution clothes, and inconvenience is cleaned after being infected with, sale lily generally all carries out artificial emasculation processing in the market, process is cumbersome, waste a large amount of manpower and materials, so cultivate without pollen lily be always Lilies breeding emphasis direction, while with excellent fancy points without pollen lily also with huge market prospects.Of the invention innovative is combined using special hybrid lily breeding, it is cultivated, without the screening of pollen lily and multiplication technique with reference to sowing, lily tissue culture propagation, lily small seed ball refrigeration vernalization, lily ball in bottle, creative breeding cycle of the shortening without pollen lily, and select the excellent no pollen lily new varieties of fancy points.
Description
Technical field
The invention belongs to the new variety selective breeding technology fields of plant, and in particular to a kind of cultivation side of no pollen lily cultivar
Method.
Background technology
Lily is Liliaceae lilium perennial herb bulb bearing plant, is world-renowned ornamental flower, in recent years, with
China's expanding economy and the raising of living standards of the people, lily have become high-grade cut-flower important in China flowers market.Extensively
It is general to apply to courtyard greening, home decoration, the fields such as red-letter day flower, medical, edible.With the continuous expansion of lily market,
Fresh flower lily, potted lily, courtyard greening lily ball are planted extensively in the area such as Southwestern China, northwest, northeast.But due to hundred
It closes pollen and is easier to pollution clothes, and inconvenience is cleaned after being infected with, sale cut-flower and potted lily generally all carry out artificial in the market
Emasculation is handled, and process is cumbersome, wastes a large amount of manpower and materials, thus cultivate without pollen lily be always Lilies breeding emphasis side
To, at the same with excellent fancy points without pollen lily also with huge market prospects.Foreign countries are to male sterile at present
The basic research that no pollen lily has some preliminary also has the report without pollen lily cultivar of only a few, but how to obtain
The type lily new varieties, foreign data are said nothing.At home, the basic research either without pollen lily, new varieties
Selection and breeding, tissue culture raising technology or related cultivation technique belong to blank.China is big as lily resource distribution center and consumption
State grasps the breeding core technology without pollen lily with great market prospect, and develops relevant lily new varieties, to me
The development of state's industry of flowers and plants kind industry innovation has huge impetus.
Invention content
To solve the deficiencies in the prior art, exploitation is shortened related with independent intellectual property right without pollen lily new varieties
Breeding cycle, the present invention on the basis of the hereditary capacity without pollen lily is grasped, it is innovative miscellaneous using special lily
Breeding combination is handed over, with reference to seeding technique in bottle, lily tissue culture propagation technology, lily small seed ball refrigeration vernalization technology, lily ball
Cultivation technique, no pollen lily screening and multiplication technique shorten the breeding cycle of no pollen lily, and it is excellent to select fancy points
It is good without pollen lily new varieties.
To achieve the above object, the technical solution adopted by the present invention is as follows：
A kind of breeding method of no pollen lily cultivar, includes the following steps：
Step A, July is in lily full-bloom stage, selection male parent oriental hybrid lily anther grain, in the greenhouse not higher than 25 DEG C
In, artificial pollination is carried out to maternal oriental hybrid lily, wraps up the column cap of maternal oriental hybrid lily after pollination with masking foil；
Step B pollinates 85-95 days, chooses the lily fruit that capsule expands, observes daily, treats capsule maturation, but dorsal suture
Capsule is removed when not cracking, then with after twice of the alcohol wipe capsule that mass concentration is 75%, it is dense that capsule is first put into quality
It spends in the mercuric chloride solution for 0.1% and sterilizes 15min, place into the first mixed liquor and sterilize 15min, later with aseptic water washing 3
Secondary, each 1min obtains lily seed explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step C, peels off step on superclean bench（2）Obtained lily seed explant takes out full with endosperm
With the lily sheet seed of visible embryo, be seeded in the first propagating culture medium and carry out aseptic seeding, intensity of illumination for 2000~
2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, carry out germination induction；
The first propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8~6.0；
Each seed after germination, is removed kind of a skin, every seed after stripping kind skin is individually transferred by step D
It is 2000~2500 lx in intensity of illumination, temperature is 25 ± 2 DEG C, and light application time is 12~14h/d to the second propagating culture medium
Under conditions of, it cultivates 30~40 days；
The second propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
0.3~0.5mg/L of 6-benzyladenine；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8~6.0；
Step E is forwarded to third propagating culture medium after step D is cultivated 30~40 days, and it is 2000 to continue in intensity of illumination
~2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, are cultivated 30~40 days；
The third propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
0.8~1.0mg/L of 6-benzyladenine
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
PH 5.8~6.0；
Step F after step E is cultivated 30~40 days, is replaced with new propagating culture medium, is trained again according to the condition of step E
It supports 30~40 days, the lily for selecting all 3~4cm of diameter of robust growth later expands seed ball, is forwarded to root media, light culture
30~40 days；
The root media of the 1L contains following component：
MS culture mediums 4.4g；
0.8~1.0 mg/L of methyl α-naphthyl acetate；
Sucrose 60g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8~6.0；
Step G after step F is cultivated 30~40 days, is replaced with new root media, is trained again according to the condition of step F
It supports 30~40 days, obtains lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, and obtained mixture is to lily seed
Ball is packed；Every lily seed ball uses 10g~16.7g mixtures；
Packaged lily seed ball is stored in -1 DEG C of freezer environment by step H, outbound after storage 90 days, later room temperature
Storage is planted down after 3 days；
Step I, after planting 2 years lowerly, it is seen that bulb is bloomed, and screens no pollen and the good lily of fancy points；
Step J after filtering out no pollen and the good lily of fancy points, after lily above-ground plant parts is withered, takes
Go out the bulb, peel off outer scale, then with twice of the alcohol wipe that mass concentration is 75%, being then first put into mass concentration is
15min is sterilized in 0.1% mercuric chloride solution, places into the first mixed liquor and sterilizes 15min, later with aseptic water washing 3 times,
Each 1min obtains lily bud scale explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step J lily bud scale explants are forwarded to the 4th propagating culture medium by step K, intensity of illumination for 2000~
2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, are cultivated 30~40 days；
The 4th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
0.2~0.3mg/L of 6-benzyladenine
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
PH 5.8~6.0；
Step L, after step K cultivate 30~40 days after, be forwarded in the 5th propagating culture medium, continue be in intensity of illumination
2000~2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, are cultivated 30~40 days；
The 5th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
0.5~0.8mg/L of 6-benzyladenine
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
PH 5.8~6.0；
Step M, after step L cultivate 30~40 days after, be forwarded in third propagating culture medium, continue be in intensity of illumination
2000~2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, cultivates 30~40 days, are forwarded to life
Root culture medium replaces a subculture in every 30~40 days, and light culture obtains no pollen lily seed ball；It is 7 according to mass ratio:3 will
Turfy soil and water mixing, obtained mixture pack no pollen lily seed ball；Every uses 10g without pollen lily seed ball
~16.7g mixtures；
Packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment by step N, outbound after storage 90 days,
Room temperature storage is planted down after 3 days later；No pollen lily original source kind is obtained plant 2 years lowerly after.
It is further preferred that the fertilizing management method after being planted under step I and step N is as follows：
（1）It after plantation, when stem-root grows 5-10mm, pours and applies 2 secondary root fertilizers, fertilising for the first time is the 8- after plantation
12 days, the 2nd fertilising was 18-22 days after plantation, and the component for applying fertilizer of taking root every time is as shown in table 1, i.e., table 1 is applied for 1 time
Fertile dosage；
Table 1
（2）At jointing stage, squaring period and the florescence of lily growth, a summer fertilizer was applied every 7 days, applies 10 times altogether, often
The component of secondary applied summer fertilizer is as shown in table 2, i.e., table 2 is 1 fertilising dosage；
Table 2
The present invention is innovative to be educated using special hybrid lily on the basis of the hereditary capacity without pollen lily is grasped
Kind combination, with reference to seeding technique, lily tissue culture propagation technology, lily small seed ball refrigeration vernalization technology, lily ball cultivation in bottle
Technology, without the screening of pollen lily and multiplication technique, creativeness shortens the breeding cycle without pollen lily, and selects fancy points
It is excellent without pollen lily new varieties.
Compared with prior art, the present invention advantage is：
1. the hybrid strain of no pollen lily offspring can be generated by being found that, which is all polliniferous normal hundred
It closes, and fancy points is preferable, but can occur male sterile no pollen lily after hybridization, and the combination occurs without pollen ratio
Rate is larger（Offspring has pollen plant：Offspring is without pollen plant ≈ 4:1）, greatly improve the selection and breeding of no pollen lily new varieties
Efficiency.
2. by aseptic seeding technology, the survival rate of filial generation seed is improved, is carried from the 75% of conventional seed planting survival rate
Height can ensure that no pollen hybrid lily progeny seed is not lost in more than 95%.
3. can solve the problems, such as that the seedling parental source of conventional breeding of method production is complicated, by the implementation of the present invention,
It is single without pollen lily seed ball source, it is easy to standardization, batch production operation, is effectively improved seed ball quality, can be carried for market
For the excellent no pollen lily original source kind of unified standard.
4. after making the breeding to 30 clones of no pollen lily seed ball by tissue culture, then expand and field cultivation of taking root, have
The survival rate for improving single offspring of effect substantially reduced the breeding cycle of no pollen lily, from the breeding of original averagely 9 years
Cycle time was to 6 years or so.
Description of the drawings
Fig. 1 is lily of the method for the present invention institute's selection and breeding without pollen lily original source kind；
Fig. 2 is the method for the present invention institute's selection and breeding without the another lily of pollen lily original source kind.
Specific embodiment
With reference to embodiment, the present invention is described in further detail.
It will be understood to those of skill in the art that the following example is merely to illustrate the present invention, and it should not be regarded as limiting this hair
Bright range.In the examples where no specific technique or condition is specified, according to the described technology of document in the art or condition
Or it is carried out according to product description.Reagents or instruments used without specified manufacturer, being can be by buying what is obtained
Conventional products.
Embodiment 1
A kind of breeding method of no pollen lily cultivar, includes the following steps：
Step A, July is in lily full-bloom stage, selection male parent oriental hybrid lily anther grain, in the greenhouse not higher than 25 DEG C
In, artificial pollination is carried out to maternal oriental hybrid lily, wraps up the column cap of maternal oriental hybrid lily after pollination with masking foil；
Step B pollinates 85 days, chooses the lily fruit that capsule expands, observes daily, treats capsule maturation, but dorsal suture does not have
Capsule is removed when having cracking, then with after twice of the alcohol wipe capsule that mass concentration is 75%, capsule is first put into mass concentration
To sterilize 15min in 0.1% mercuric chloride solution, place into the first mixed liquor and sterilize 15min, later with aseptic water washing 3
Secondary, each 1min obtains lily seed explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step C, peels off step on superclean bench（2）Obtained lily seed explant takes out full with endosperm
It with the lily sheet seed of visible embryo, is seeded in the first propagating culture medium and carries out aseptic seeding, be 2000lx in intensity of illumination,
Temperature is 25 ± 2 DEG C, under conditions of light application time is 12h/d, carries out germination induction；
The first propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.8；
Each seed after germination, is removed kind of a skin, every seed after stripping kind skin is individually transferred by step D
It is 2000 lx in intensity of illumination to the second propagating culture medium, temperature is 25 ± 2 DEG C, under conditions of light application time is 12h/d, training
It supports 30 days；
The second propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
6-benzyladenine 0.3mg/L；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.8；
Step E is forwarded to third propagating culture medium after step D is cultivated 30 days, and it is 2000lx to continue in intensity of illumination, warm
It is 25 ± 2 DEG C to spend, and under conditions of light application time is 12h/d, is cultivated 30 days；
The third propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.8mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 5.8；
Step F after step E is cultivated 30 days, is replaced with new propagating culture medium, 30 is further cultured for according to the condition of step E
My god, the lily for selecting all diameter 3cm of robust growth later expands seed ball, is forwarded to root media, light culture 30 days；
The root media of the 1L contains following component：
MS culture mediums 4.4g；
Methyl α-naphthyl acetate 0.8mg/L；
Sucrose 60g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.8；
Step G after step F is cultivated 30 days, is replaced with new root media, 30 is further cultured for according to the condition of step F
My god, obtain lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, and obtained mixture wraps lily seed ball
Dress；Every lily seed ball uses 10g mixtures；
Packaged lily seed ball is stored in -1 DEG C of freezer environment by step H, outbound after storage 90 days, later room temperature
Storage is planted down after 3 days；
Step I, after planting 2 years lowerly, it is seen that bulb is bloomed, and screens no pollen and the good lily of fancy points；
Step J after filtering out no pollen and the good lily of fancy points, after lily above-ground plant parts is withered, takes
Go out the bulb, peel off outer scale, then with twice of the alcohol wipe that mass concentration is 75%, being then first put into mass concentration is
15min is sterilized in 0.1% mercuric chloride solution, places into the first mixed liquor and sterilizes 15min, later with aseptic water washing 3 times,
Each 1min obtains lily bud scale explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step J lily bud scale explants are forwarded to the 4th propagating culture medium by step K, are 2000lx in intensity of illumination, warm
It is 25 ± 2 DEG C to spend, and under conditions of light application time is 12h/d, is cultivated 30 days；
The 4th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.2mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 5.8；
Step L after step K is cultivated 30 days, is forwarded in the 5th propagating culture medium, and it is 2000 to continue in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12h/d, are cultivated 30 days；
The 5th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.5mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 5.8；
Step M, after step L cultivate 30 days after, be forwarded in third propagating culture medium, continue be in intensity of illumination
2000lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12h/d, cultivates 30 days, are forwarded to root media, every 30 days
A subculture is replaced, light culture obtains no pollen lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, obtain
Mixture no pollen lily seed ball is packed；Every uses 10g mixtures without pollen lily seed ball；
Packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment by step N, outbound after storage 90 days,
Room temperature storage is planted down after 3 days later；No pollen lily original source kind is obtained plant 2 years lowerly after.
Wherein, the fertilizing management method after being planted under step I and step N is as follows：
（1）It after plantation, when stem-root grows 5-10mm, pours and applies 2 secondary root fertilizers, fertilising for the first time is 8 after plantation
My god, the 2nd fertilising is 18 days after plantation, and the component for applying fertilizer of taking root every time is as shown in the table, i.e., amount is 1 used in table
Secondary fertilising dosage；
Table 1
（2）At jointing stage, squaring period and the florescence of lily growth, a summer fertilizer was applied every 7 days, applies 10 times altogether, often
The component of secondary applied summer fertilizer is as shown in the table, i.e., amount is 1 fertilising dosage used in table；
Table 2
Embodiment 2
A kind of breeding method of no pollen lily cultivar, includes the following steps：
Step A, July is in lily full-bloom stage, selection male parent oriental hybrid lily anther grain, in the greenhouse not higher than 25 DEG C
In, artificial pollination is carried out to maternal oriental hybrid lily, wraps up the column cap of maternal oriental hybrid lily after pollination with masking foil；
Step B pollinates 95 days, chooses the lily fruit that capsule expands, observes daily, treats capsule maturation, but dorsal suture does not have
Capsule is removed when having cracking, then with after twice of the alcohol wipe capsule that mass concentration is 75%, capsule is first put into mass concentration
To sterilize 15min in 0.1% mercuric chloride solution, place into the first mixed liquor and sterilize 15min, later with aseptic water washing 3
Secondary, each 1min obtains lily seed explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step C, peels off step on superclean bench（2）Obtained lily seed explant takes out full with endosperm
It with the lily sheet seed of visible embryo, is seeded in the first propagating culture medium and carries out aseptic seeding, be 2500lx in intensity of illumination,
Temperature is 25 ± 2 DEG C, under conditions of light application time is 14h/d, carries out germination induction；
The first propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 6.0；
Each seed after germination, is removed kind of a skin, every seed after stripping kind skin is individually transferred by step D
It is 2500 lx in intensity of illumination to the second propagating culture medium, temperature is 25 ± 2 DEG C, under conditions of light application time is 14h/d, training
It supports 40 days；
The second propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
6-benzyladenine 0.5mg/L；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 6.0；
Step E is forwarded to third propagating culture medium after step D is cultivated 40 days, and it is 2500 lx to continue in intensity of illumination,
Temperature is 25 ± 2 DEG C, under conditions of light application time is 14h/d, is cultivated 40 days；
The third propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 1.0mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 6.0；
Step F after step E is cultivated 40 days, is replaced with new propagating culture medium, 40 is further cultured for according to the condition of step E
My god, the lily for selecting all diameter 4cm of robust growth later expands seed ball, is forwarded to root media, light culture 40 days；
The root media of the 1L contains following component：
MS culture mediums 4.4g；
1.0 mg/L of methyl α-naphthyl acetate；
Sucrose 60g/L；
Agarose 6g/L；
Deionized water surplus；
pH 56.0；
Step G after step F is cultivated 40 days, is replaced with new root media, 40 is further cultured for according to the condition of step F
My god, obtain lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, and obtained mixture wraps lily seed ball
Dress；Every lily seed ball uses 16.7g mixtures；
Packaged lily seed ball is stored in -1 DEG C of freezer environment by step H, outbound after storage 90 days, later room temperature
Storage is planted down after 3 days；
Step I, after planting 2 years lowerly, it is seen that bulb is bloomed, and screens no pollen and the good lily of fancy points；
Step J after filtering out no pollen and the good lily of fancy points, after lily above-ground plant parts is withered, takes
Go out the bulb, peel off outer scale, then with twice of the alcohol wipe that mass concentration is 75%, being then first put into mass concentration is
15min is sterilized in 0.1% mercuric chloride solution, places into the first mixed liquor and sterilizes 15min, later with aseptic water washing 3 times,
Each 1min obtains lily bud scale explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step J lily bud scale explants are forwarded to the 4th propagating culture medium by step K, are 2500 lx in intensity of illumination,
Temperature is 25 ± 2 DEG C, under conditions of light application time is 14h/d, is cultivated 40 days；
The 4th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.3mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 6.0；
Step L after step K is cultivated 40 days, is forwarded in the 5th propagating culture medium, and it is 2500 to continue in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 14h/d, are cultivated 40 days；
The 5th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.8mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 6.0；
Step M after step L is cultivated 40 days, is forwarded in third propagating culture medium, and it is 2500 to continue in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 14h/d, are cultivated 40 days, are forwarded to root media, are replaced within every 40 days
One subculture, light culture obtain no pollen lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, and what is obtained is mixed
Object is closed to pack no pollen lily seed ball；Every uses 16.7g mixtures without pollen lily seed ball；
Packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment by step N, outbound after storage 90 days,
Room temperature storage is planted down after 3 days later；No pollen lily original source kind is obtained plant 2 years lowerly after.
Wherein, the fertilizing management method after being planted under step I and step N is as follows：
（1）It after plantation, when stem-root grows 5-10mm, pours and applies 2 secondary root fertilizers, fertilising for the first time is 12 after plantation
My god, the 2nd fertilising is 22 days after plantation, and the component for applying fertilizer of taking root every time is as shown in the table, i.e., amount is 1 used in table
Secondary fertilising dosage；
Table 1
（2）At jointing stage, squaring period and the florescence of lily growth, a summer fertilizer was applied every 7 days, applies 10 times altogether, often
The component of secondary applied summer fertilizer is as shown in the table, i.e., amount is 1 fertilising dosage used in table；
Table 2
Embodiment 3
A kind of breeding method of no pollen lily cultivar, includes the following steps：
Step A, July is in lily full-bloom stage, selection male parent oriental hybrid lily anther grain, in the greenhouse not higher than 25 DEG C
In, artificial pollination is carried out to maternal oriental hybrid lily, wraps up the column cap of maternal oriental hybrid lily after pollination with masking foil；
Step B pollinates 90 days, chooses the lily fruit that capsule expands, observes daily, treats capsule maturation, but dorsal suture does not have
Capsule is removed when having cracking, then with after twice of the alcohol wipe capsule that mass concentration is 75%, capsule is first put into mass concentration
To sterilize 15min in 0.1% mercuric chloride solution, place into the first mixed liquor and sterilize 15min, later with aseptic water washing 3
Secondary, each 1min obtains lily seed explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step C, peels off step on superclean bench（2）Obtained lily seed explant takes out full with endosperm
It with the lily sheet seed of visible embryo, is seeded in the first propagating culture medium and carries out aseptic seeding, be 2200 in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 13h/d, carry out germination induction；
The first propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.9；
Each seed after germination, is removed kind of a skin by step D, will be removed every seed after kind of skin and is individually forwarded to the
Two propagating culture mediums are 2200 lx in intensity of illumination, and temperature is 25 ± 2 DEG C, under conditions of light application time is 13h/d, is cultivated 35 days；
The second propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
6-benzyladenine 0.4mg/L；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.9；
Step E is forwarded to third propagating culture medium after step D is cultivated 35 days, and it is 2200 lx to continue in intensity of illumination,
Temperature is 25 ± 2 DEG C, under conditions of light application time is 13h/d, is cultivated 35 days；
The third propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.93mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 5.9；
Step F after step E is cultivated 35 days, is replaced with new propagating culture medium, 35 is further cultured for according to the condition of step E
My god, the lily for selecting all diameter 3.5cm of robust growth later expands seed ball, is forwarded to root media, light culture 35 days；
The root media of the 1L contains following component：
MS culture mediums 4.4g；
0.9 mg/L of methyl α-naphthyl acetate；
Sucrose 60g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.9；
Step G after step F is cultivated 35 days, is replaced with new root media, 35 is further cultured for according to the condition of step F
My god, obtain lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, and obtained mixture wraps lily seed ball
Dress；Every lily seed ball uses 13g mixtures；
Packaged lily seed ball is stored in -1 DEG C of freezer environment by step H, outbound after storage 90 days, later room temperature
Storage is planted down after 3 days；
Step I, after planting 2 years lowerly, it is seen that bulb is bloomed, and screens no pollen and the good lily of fancy points；
Step J after filtering out no pollen and the good lily of fancy points, after lily above-ground plant parts is withered, takes
Go out the bulb, peel off outer scale, then with twice of the alcohol wipe that mass concentration is 75%, being then first put into mass concentration is
15min is sterilized in 0.1% mercuric chloride solution, places into the first mixed liquor and sterilizes 15min, later with aseptic water washing 3 times,
Each 1min obtains lily bud scale explant；
The preparation method of first mixed liquor is to be added dropwise to have in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
5 drop Tween-20s, be uniformly mixed to get；
Step J lily bud scale explants are forwarded to the 4th propagating culture medium by step K, are 2200 lx in intensity of illumination,
Temperature is 25 ± 2 DEG C, under conditions of light application time is 13h/d, is cultivated 35 days；
The 4th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.25mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 5.9；
Step L after step K is cultivated 35 days, is forwarded in the 5th propagating culture medium, and it is 2200 to continue in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 13h/d, are cultivated 35 days；
The 5th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.6mg/L
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
pH 5.9；
Step M after step L is cultivated 35 days, is forwarded in third propagating culture medium, and it is 2200 to continue in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 13h/d, are cultivated 35 days, are forwarded to root media, are replaced within every 35 days
One subculture, light culture obtain no pollen lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, and what is obtained is mixed
Object is closed to pack no pollen lily seed ball；Every uses 13g mixtures without pollen lily seed ball；
Packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment by step N, outbound after storage 90 days,
Room temperature storage is planted down after 3 days later；No pollen lily original source kind is obtained plant 2 years lowerly after.
Wherein, the fertilizing management method after being planted under step I and step N is as follows：
（1）It after plantation, when stem-root grows 5-10mm, pours and applies 2 secondary root fertilizers, fertilising for the first time is 10 after plantation
My god, the 2nd fertilising is 20 days after plantation, and the component for applying fertilizer of taking root every time is as shown in the table, i.e., amount is 1 used in table
Secondary fertilising dosage；
Table 1
（2）At jointing stage, squaring period and the florescence of lily growth, a summer fertilizer was applied every 7 days, applies 10 times altogether, often
The component of secondary applied summer fertilizer is as shown in the table, i.e., amount is 1 fertilising dosage used in table；
Table 2
The present embodiment obtained no pollen lily original source kind in 2015, as shown in Figure 1 and 2.
It is oriental hybrid lily Xi Bailun that the present embodiment, which is chosen maternal, and male parent is oriental hybrid lily Marco Polo.
Obtained by the present embodiment without pollen lily original source kind offspring's kind characteristic：1. male sterility is without pollen.2. Dan Zhihua
Head number 4-5.3. cut-flower production period 100-120 days.4. pattern：Inner valves, outer valve outer rim are pink, inner white.5. flower-shape：It is star-like,
Outer valve outer rim big ripple.
Basic principle, main feature and the advantages of the present invention of the present invention has been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent thereof.
Claims (2)
1. a kind of breeding method of no pollen lily cultivar, which is characterized in that include the following steps：
Step A, July in lily full-bloom stage, chooses male parent oriental hybrid lily anther grain, right in the greenhouse not higher than 25 DEG C
Maternal oriental hybrid lily carries out artificial pollination, wraps up the column cap of maternal oriental hybrid lily after pollination with masking foil；It is described maternal for east
Lily Xi Bailun, male parent are oriental hybrid lily Marco Polo；
Step B pollinates 85-95 days, chooses the lily fruit that capsule expands, observes daily, treats capsule maturation, but dorsal suture does not have
Capsule is removed during cracking, then with after twice of the alcohol wipe capsule that mass concentration is 75%, capsule is first put into mass concentration is
15min is sterilized in 0.1% mercuric chloride solution, places into the first mixed liquor and sterilizes 15min, later with aseptic water washing 3 times,
Each 1min obtains lily seed explant；
The preparation method of first mixed liquor is to be added dropwise to have 5 drops in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
Tween-20, be uniformly mixed to get；
Step C, peels off step on superclean bench（2）Obtained lily seed explant, taking out full has endosperm and can
See the lily sheet seed of embryo, be seeded in the first propagating culture medium and carry out aseptic seeding, be 2000~2500 in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, carry out germination induction；
The first propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8~6.0；
Each seed after germination, is removed kind of a skin by step D, will be removed every seed after kind of skin and is individually forwarded to the
Two propagating culture mediums are 2000~2500 lx in intensity of illumination, and temperature is 25 ± 2 DEG C, and light application time is the item of 12~14h/d
Under part, cultivate 30~40 days；
The second propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
0.3~0.5mg/L of 6-benzyladenine；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8~6.0；
Step E is forwarded to third propagating culture medium after step D is cultivated 30~40 days, continue intensity of illumination for 2000~
2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, are cultivated 30~40 days；
The third propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
0.8~1.0mg/L of 6-benzyladenine
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
PH 5.8~6.0；
Step F after step E is cultivated 30~40 days, is replaced with new propagating culture medium, 30 are further cultured for according to the condition of step E~
40 days, the lily for selecting all 3~4cm of diameter of robust growth later expanded seed ball, was forwarded to root media, light culture 30~40 days；
The root media of the 1L contains following component：
MS culture mediums 4.4g；
0.8~1.0 mg/L of methyl α-naphthyl acetate；
Sucrose 60g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8~6.0；
Step G after step F is cultivated 30~40 days, is replaced with new root media, 30 is further cultured for according to the condition of step F
~40 days, obtain lily seed ball；It is 7 according to mass ratio:3 mix turfy soil and water, obtained mixture to lily seed ball into
Row packaging；Every lily seed ball uses 10g~16.7g mixtures；
Packaged lily seed ball is stored in -1 DEG C of freezer environment by step H, outbound after storage 90 days, later room temperature storage 3
It is planted down after it；
Step I, after planting 2 years lowerly, it is seen that bulb is bloomed, and screens no pollen and the good lily of fancy points；
Step J, after filtering out no pollen and the good lily of fancy points, after lily above-ground plant parts is withered, taking out should
Bulb peels off outer scale, and then with twice of the alcohol wipe that mass concentration is 75%, it is 0.1% to be then first put into mass concentration
Mercuric chloride solution in sterilize 15min, place into the first mixed liquor and sterilize 15min, later with aseptic water washing 3 times, every time
1min obtains lily bud scale explant；
The preparation method of first mixed liquor is to be added dropwise to have 5 drops in the liquor natrii hypochloritis that every 100ml mass concentrations are 2%
Tween-20, be uniformly mixed to get；
Step J lily bud scale explants are forwarded to the 4th propagating culture medium by step K, are 2000~2500 in intensity of illumination
Lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, are cultivated 30~40 days；
The 4th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
0.2~0.3mg/L of 6-benzyladenine
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
PH 5.8~6.0；
Step L after step K is cultivated 30~40 days, is forwarded in the 5th propagating culture medium, continue intensity of illumination for 2000~
2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, are cultivated 30~40 days；
The 5th propagating culture medium of the 1L contains following component：
MS culture mediums 4.4g
Methyl α-naphthyl acetate 0.1mg/L
0.5~0.8mg/L of 6-benzyladenine
Sucrose 30g/L
Agarose 6g/L
Deionized water surplus；
PH 5.8~6.0；
Step M after step L is cultivated 30~40 days, is forwarded in third propagating culture medium, continue intensity of illumination for 2000~
2500 lx, temperature are 25 ± 2 DEG C, under conditions of light application time is 12~14h/d, cultivates 30~40 days, are forwarded to culture of rootage
Base replaces a subculture in every 30~40 days, and light culture obtains no pollen lily seed ball；It is 7 according to mass ratio:3 by turfy soil
It is mixed with water, obtained mixture packs no pollen lily seed ball；Every without pollen lily seed ball using 10g~
16.7g mixtures；
Packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment by step N, outbound after storage 90 days, later
Room temperature storage is planted down after 3 days；No pollen lily original source kind is obtained plant 2 years lowerly after.
2. the breeding method of no pollen lily cultivar according to claim 1, which is characterized in that under step I and step N
Fertilizing management method after plantation is as follows：
（1）It after plantation, when stem-root grows 5-10mm, pours and applies 2 secondary root fertilizers, fertilising for the first time is the 8-12 after plantation
My god, the 2nd fertilising is 18-22 days after plantation, and the component for applying fertilizer of taking root based on kg/ mus every time is：Calcium nitrate 4.9, nitre
Sour ammonium 3.0, potassium nitrate 3.0, magnesium sulfate 5.5, potassium chloride 0.14, potassium dihydrogen phosphate 8.5, phosphoric acid 0.8, boric acid 0.25；
（2）At jointing stage, squaring period and the florescence of lily growth, a summer fertilizer was applied every 7 days, applies 10 times, presses every time altogether
It counts for kg/ mus and applies the component of summer fertilizer and be：Organic fertilizer 15, calcium nitrate 4.7, ammonium nitrate 0.24, potassium nitrate 3.0, urea 0.5,
Magnesium sulfate 2.2, potassium chloride 0.1, boric acid 0.04, tartaric acid 0.16, ferrous sulfate 0.17, manganese sulfate 0.05, zinc sulfate 0.03, sulphur
Sour copper 0.005, nickel sulfate 0.003, ammonium molybdate 0.04, potassium dihydrogen phosphate 0.3.
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| CN107182772A (en) * | 2017-06-08 | 2017-09-22 | 南京农业大学 | Method and application of a kind of efficient seed selection without pollen contamination chrysanthemum |
| CN109380102A (en) * | 2017-08-10 | 2019-02-26 | 辽宁省农业科学院 | A kind of modularization soilless cultivation new technology for lily bud scale bulb-lets breeding |
| CN107828909A (en) * | 2017-11-15 | 2018-03-23 | 云南省农业科学院花卉研究所 | A kind of male sterile method of Markers for Detection oriental hybrid lily special group |
| CN107771638A (en) * | 2017-11-23 | 2018-03-09 | 蚌埠宏瑞园林有限公司 | A kind of lily culture method for improving lily bulb medical value |
| CN113196981A (en) * | 2021-04-28 | 2021-08-03 | 北京市辐射中心 | Planting pot and planting method for lily hybrid progeny bulbs |
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