CN106417000A - Pollen-free lily variety breeding method - Google Patents
Pollen-free lily variety breeding method Download PDFInfo
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- CN106417000A CN106417000A CN201610852456.XA CN201610852456A CN106417000A CN 106417000 A CN106417000 A CN 106417000A CN 201610852456 A CN201610852456 A CN 201610852456A CN 106417000 A CN106417000 A CN 106417000A
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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 invention relates to a pollen-free lily variety breeding method and belongs to the technical field of breeding of new plant varieties. Lily is a main commercial flower in the world, is widely planted around China and occupies huge market shares. However, due to the fact that lily pollen easily stains clothes, the clothes stained with pollen is inconvenient to clean, the lily sold in the market generally undergoes artificial castration processing, the process is complicated and a large number of manpower and material resources is wasted, a bred pollen-free lily is always the key direction of lily breeding, and meanwhile the pollen-free lily having excellent ornamental characteristics also has a huge market prospect. The method creatively utilizes special lily crossbreeding combination, combines with in-bottle sowing, lily tissue culture and breeding, cold storage and vernalization of small lily bulblets, lily seed ball cultivation, pollen-free lily screening and propagation expanding technologies, a breeding period of the pollen-free lily is creatively shortened, and the new pollen-free lily variety having excellent ornamental characteristics is bred.
Description
Technical field
The invention belongs to the new variety selective breeding technology field of plant is 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 has become important high-grade cut-flower in China flowers market.Extensively
General apply to courtyard greening, home decoration, red-letter day flower, the field such as medical, edible.With the continuous expansion of lily market,
Fresh flower lily, potted lily, courtyard greening lily ball are extensively planted in the area such as Southwestern China, northwest, northeast.But due to hundred
Close pollen and be easier to pollution clothes, and cleaning inconvenience after contamination, cut-flower is sold in market and potted lily is typically all carried out manually
Emasculation is processed, and process is loaded down with trivial details, wastes substantial amounts of manpower and materials, so cultivating the emphasis side that no pollen lily is always Lilies breeding
To the no pollen lily with excellent fancy points also has huge market prospects.At present abroad to male sterile
No pollen lily has some preliminary basic research, also has the report of the no pollen lily cultivar of only a few, but how to obtain
The type lily new varieties, foreign data is said nothing.At home, the either no basic research of pollen lily, new varieties
Seed selection, tissue culture raising technology, or related cultivation technique broadly falls into blank.China is big as lily resource distribution center and consumption
State, grasps the breeding core technology of the no pollen lily with great market prospect, and develops related lily new varieties, to me
The development of state's industry of flowers and plants kind industry innovation has huge impetus.
Content of the invention
For solving the deficiencies in the prior art, exploitation has the no pollen lily new varieties of independent intellectual property right, shortens related
Breeding cycle, the present invention grasping no on the basis of the hereditary capacity of pollen lily, and the lily that the utilization of novelty is special is miscellaneous
Hand over breeding combination, in conjunction with seeding technique in bottle, lily tissue culture propagation technology, lily small seed ball refrigerates vernalization technology, lily ball
The screening of cultivation technique, no pollen lily and multiplication technique, shorten the breeding cycle of no pollen lily, and it are excellent to select fancy points
Good no pollen lily new varieties.
For achieving the above object, the technical solution used in the present invention is as follows：
A kind of breeding method of no pollen lily cultivar, comprises the steps：
Step A, July, in lily full-bloom stage, chooses male parent oriental hybrid lily flower pesticide grain, in not higher than 25 DEG C of warmhouse booth, right
Maternal oriental hybrid lily carries out artificial pollination, 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, daily observation, treats that capsule is ripe, but dorsal suture does not have
Capsule is taken off, after twice of the alcohol wipe capsule being then 75% with mass concentration, capsule first being put into mass concentration is during cracking
Sterilize in 0.1% mercuric chloride solution 15min, places in the first mixed liquor sterilization 15min, uses aseptic water washing 3 times afterwards,
1min every time, obtains lily seed explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step C, strip off step on superclean bench（2）The lily seed explant obtaining, takes out and 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 is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, carries out germination induction；
First propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8～6.0；
Step D, after germination, each seed is peeled off kind of a skin, and the every seed peeled off after kind of skin 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 bar of 12～14h/d
Under part, cultivate 30～40 days；
Second propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
6-benzyladenine 0.3～0.5mg/L；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8～6.0；
Step E, is forwarded to the 3rd propagating culture medium after step D is cultivated 30～40 days, continue intensity of illumination be 2000～
2500 lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days；
3rd propagating culture medium of described 1L contains following component：
MS culture medium 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.8～1.0mg/L
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, the condition according to step E is further cultured for 30
～40 days, the lily selecting all footpath 3～4cm of robust growth afterwards expanded seed ball, was forwarded to root media, and light culture 30～
40 days；
The root media of described 1L contains following component：
MS culture medium 4.4g；
Methyl α-naphthyl acetate 0.8～1.0 mg/L；
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, the condition according to step F is further cultured for 30
～40 days, obtain lily seed ball；It is 7 according to mass ratio:Turfy soil and water mixing, the mixture obtaining are entered by 3 to lily seed ball
Row packaging；Every lily seed ball uses 10g～16.7g mixture；
Step H, packaged lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, room temperature storage 3 afterwards
Plant down after it；
Step I, it is seen that bulb is bloomed lowerly after planting 2 years, screening no pollen and the good lily of fancy points；
Step J, filters out no after pollen and the good lily of fancy points, after lily above-ground plant parts is withered, taking out should
Bulb, peels off outer scale, twice of the alcohol wipe being then 75% with mass concentration, and then first putting into mass concentration is 0.1%
Mercuric chloride solution in sterilize 15min, place in the first mixed liquor sterilization 15min, use aseptic water washing 3 times afterwards, every time
1min, obtains lily bud scale explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step K, step J lily bud scale explant is forwarded to the 4th propagating culture medium, is 2000～2500 in intensity of illumination
Lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days；
4th propagating culture medium of described 1L contains following component：
MS culture medium 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.2～0.3mg/L
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 intensity of illumination be 2000～
2500 lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days；
5th propagating culture medium of described 1L contains following component：
MS culture medium 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.5～0.8mg/L
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 the 3rd propagating culture medium, continue intensity of illumination be 2000～
2500 lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days, is forwarded to culture of rootage
Base, changes a subculture, light culture in every 30～40 days, obtains no pollen lily seed ball；It is 7 according to mass ratio:3 by turfy soil
Mix with water, the mixture obtaining is packed to no pollen lily seed ball；Every no pollen lily seed ball use 10g～
16.7g mixture；
Step N, packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, afterwards
Room temperature storage was planted down after 3 days；Obtain no pollen lily former source kind after planting 2 years lowerly.
It is further preferred that the fertilizing management method after planting under step I and step N is as follows：
（1）After plantation, when stem-root grows 5-10mm, pour and apply 2 secondary root manures, fertilising for the first time is the 8-12 after plantation
My god, the 2nd fertilising is 18-22 days after plantation, is applied the component of fertilizer of taking root every time as shown in table 1, and that is, table 1 is 1 fertilising
Consumption；
Table 1
（2）At jointing stage, squaring period and the florescence growing in lily, applied a summer fertilizer every 7 days, apply altogether 10 times, each institute
Apply the component of summer fertilizer as shown in table 2, that is, table 2 is 1 fertilising consumption；
Table 2
The present invention grasping no on the basis of the hereditary capacity of pollen lily, the special hybrid lily breeding group of the utilization of novelty
Close, in conjunction with seeding technique in bottle, lily tissue culture propagation technology, lily small seed ball refrigeration vernalization technology, lily ball cultivation technique,
The no screening of pollen lily and multiplication technique, the creative breeding cycle shortening no pollen lily, and it is excellent to select fancy points
No pollen lily new varieties.
Compared with prior art, its advantage is the present invention：
1. it is found that the hybrid strain that can produce no pollen lily offspring, this two parent is polliniferous normal lily, and
Preferably, but can male sterile no pollen lily after hybridization in fancy points, and this combination appearance no pollen ratio is relatively
Greatly（Offspring has pollen plant：Offspring no pollen plant ≈ 4:1）, greatly improve the seed selection effect of no pollen lily new varieties
Rate.
2. pass through aseptic seeding technology, improve the survival rate of filial generation seed, carry from the 75% of conventional seed planting survival rate
Height, to more than 95%, ensure that no pollen hybrid lily progeny seed does not run off.
3. can solve the problem that the problem of the seedling parental source complexity that conventional breeding of method produces, by the enforcement of the present invention,
No pollen lily seed ball is originated single it is easy to standardization, batch production operate, and is effectively improved seed ball quality, can carry for market
Excellent no pollen lily former source kind for unified standard.
4. after tissue culture makes no pollen lily seed ball breed 30 clone, then the field cultivation that expands and take root, have
The survival rate that improve single offspring of effect, substantially reduces the breeding cycle of no pollen lily, from the breeding of average 9 years originally
Cycle time was to 6 years about.
Brief description
Fig. 1 is the lily of the inventive method institute seed selection no pollen lily former source kind；
Fig. 2 is the inventive method institute seed selection no another lily of pollen lily former 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 should not be regarded as limiting this
Bright scope.Unreceipted particular technique or condition person in embodiment, according to the technology described by document in the art or condition
Or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, are and can pass through what purchase obtain
Conventional products.
Embodiment 1
A kind of breeding method of no pollen lily cultivar, comprises the steps：
Step A, July, in lily full-bloom stage, chooses male parent oriental hybrid lily flower pesticide grain, in not higher than 25 DEG C of warmhouse booth, right
Maternal oriental hybrid lily carries out artificial pollination, 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, daily observation, treats that capsule is ripe, but dorsal suture is not opened
Capsule is taken off, after twice of the alcohol wipe capsule being then 75% with mass concentration, capsule first being put into mass concentration is when splitting
Sterilize in 0.1% mercuric chloride solution 15min, places in the first mixed liquor sterilization 15min, uses aseptic water washing 3 times afterwards,
1min every time, obtains lily seed explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step C, strip off step on superclean bench（2）The lily seed explant obtaining, takes out and 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 2000lx in intensity of illumination, temperature
For 25 ± 2 DEG C, under conditions of light application time is 12h/d, carry out germination induction；
First propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.8；
Step D, after germination, each seed is peeled off kind of a skin, and the every seed peeled off after kind of skin is individually forwarded to the
Two propagating culture mediums, are 2000 lx in intensity of illumination, and temperature is 25 ± 2 DEG C, under conditions of light application time is 12h/d, cultivate 30
My god；
Second propagating culture medium of described 1L contains following component：
MS culture medium 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 the 3rd propagating culture medium, continuing in intensity of illumination is 2000lx, and temperature is after step D is cultivated 30 days
25 ± 2 DEG C, under conditions of light application time is 12h/d, cultivate 30 days；
3rd propagating culture medium of described 1L contains following component：
MS culture medium 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, the condition according to step E is further cultured for 30 days, it
The lily selecting all footpath 3cm of robust growth afterwards expands seed ball, is forwarded to root media, light culture 30 days；
The root media of described 1L contains following component：
MS culture medium 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, the condition according to step F is further cultured for 30 days, obtains
To lily seed ball；It is 7 according to mass ratio:Turfy soil and water mixing, the mixture obtaining are packed by 3 to lily seed ball；Often
Grain lily seed ball uses 10g mixture；
Step H, packaged lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, room temperature storage 3 afterwards
Plant down after it；
Step I, it is seen that bulb is bloomed lowerly after planting 2 years, screening no pollen and the good lily of fancy points；
Step J, filters out no after pollen and the good lily of fancy points, after lily above-ground plant parts is withered, taking out should
Bulb, peels off outer scale, twice of the alcohol wipe being then 75% with mass concentration, and then first putting into mass concentration is 0.1%
Mercuric chloride solution in sterilize 15min, place in the first mixed liquor sterilization 15min, use aseptic water washing 3 times afterwards, every time
1min, obtains lily bud scale explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step K, step J lily bud scale explant is forwarded to the 4th propagating culture medium, is 2000lx in intensity of illumination, and temperature is
25 ± 2 DEG C, under conditions of light application time is 12h/d, cultivate 30 days；
4th propagating culture medium of described 1L contains following component：
MS culture medium 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 continuing in intensity of illumination is 2000 lx, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 12h/d, cultivate 30 days；
5th propagating culture medium of described 1L contains following component：
MS culture medium 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 is cultivated 30 days, is forwarded in the 3rd propagating culture medium, continuing in intensity of illumination is 2000lx, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 12h/d, cultivates 30 days, be forwarded to root media, change once within every 30 days
Culture medium, light culture, obtain no pollen lily seed ball；It is 7 according to mass ratio:Turfy soil and water are mixed by 3, the mixture obtaining
No pollen lily seed ball is packed；Every no pollen lily seed ball uses 10g mixture；
Step N, packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, afterwards
Room temperature storage was planted down after 3 days；Obtain no pollen lily former source kind after planting 2 years lowerly.
Wherein, the fertilizing management method after planting under step I and step N is as follows：
（1）After plantation, when stem-root grows 5-10mm, pour and apply 2 secondary root manures, fertilising for the first time is 8 days after plantation, the
2 fertilisings are 18 days after plantation, and the component being applied fertilizer of taking root every time is as shown in the table, and that is, amount used in table is applied for 1 time
Fertile consumption；
Table 1
（2）At jointing stage, squaring period and the florescence growing in lily, applied a summer fertilizer every 7 days, apply altogether 10 times, each institute
The component applying summer fertilizer is as shown in the table, and that is, amount used in table is 1 fertilising consumption；
Table 2
Embodiment 2
A kind of breeding method of no pollen lily cultivar, comprises the steps：
Step A, July, in lily full-bloom stage, chooses male parent oriental hybrid lily flower pesticide grain, in not higher than 25 DEG C of warmhouse booth, right
Maternal oriental hybrid lily carries out artificial pollination, 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, daily observation, treats that capsule is ripe, but dorsal suture is not opened
Capsule is taken off, after twice of the alcohol wipe capsule being then 75% with mass concentration, capsule first being put into mass concentration is when splitting
Sterilize in 0.1% mercuric chloride solution 15min, places in the first mixed liquor sterilization 15min, uses aseptic water washing 3 times afterwards,
1min every time, obtains lily seed explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step C, strip off step on superclean bench（2）The lily seed explant obtaining, takes out and 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 2500lx in intensity of illumination, temperature
For 25 ± 2 DEG C, under conditions of light application time is 14h/d, carry out germination induction；
First propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 6.0；
Step D, after germination, each seed is peeled off kind of a skin, and the every seed peeled off after kind of skin is individually forwarded to the
Two propagating culture mediums, are 2500 lx in intensity of illumination, and temperature is 25 ± 2 DEG C, under conditions of light application time is 14h/d, cultivate 40
My god；
Second propagating culture medium of described 1L contains following component：
MS culture medium 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 the 3rd propagating culture medium after step D is cultivated 40 days, and continuing in intensity of illumination is 2500 lx, temperature
For 25 ± 2 DEG C, under conditions of light application time is 14h/d, cultivate 40 days；
3rd propagating culture medium of described 1L contains following component：
MS culture medium 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, the condition according to step E is further cultured for 40 days, it
The lily selecting all footpath 4cm of robust growth afterwards expands seed ball, is forwarded to root media, light culture 40 days；
The root media of described 1L contains following component：
MS culture medium 4.4g；
Methyl α-naphthyl acetate 1.0 mg/L；
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, the condition according to step F is further cultured for 40 days, obtains
To lily seed ball；It is 7 according to mass ratio:Turfy soil and water mixing, the mixture obtaining are packed by 3 to lily seed ball；Often
Grain lily seed ball uses 16.7g mixture；
Step H, packaged lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, room temperature storage 3 afterwards
Plant down after it；
Step I, it is seen that bulb is bloomed lowerly after planting 2 years, screening no pollen and the good lily of fancy points；
Step J, filters out no after pollen and the good lily of fancy points, after lily above-ground plant parts is withered, taking out should
Bulb, peels off outer scale, twice of the alcohol wipe being then 75% with mass concentration, and then first putting into mass concentration is 0.1%
Mercuric chloride solution in sterilize 15min, place in the first mixed liquor sterilization 15min, use aseptic water washing 3 times afterwards, every time
1min, obtains lily bud scale explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step K, step J lily bud scale explant is forwarded to the 4th propagating culture medium, is 2500 lx in intensity of illumination, temperature
For 25 ± 2 DEG C, under conditions of light application time is 14h/d, cultivate 40 days；
4th propagating culture medium of described 1L contains following component：
MS culture medium 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 continuing in intensity of illumination is 2500 lx, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 14h/d, cultivate 40 days；
5th propagating culture medium of described 1L contains following component：
MS culture medium 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 the 3rd propagating culture medium, and continuing in intensity of illumination is 2500 lx, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 14h/d, cultivates 40 days, be forwarded to root media, change once within every 40 days
Culture medium, light culture, obtain no pollen lily seed ball；It is 7 according to mass ratio:Turfy soil and water are mixed by 3, the mixture obtaining
No pollen lily seed ball is packed；Every no pollen lily seed ball uses 16.7g mixture；
Step N, packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, afterwards
Room temperature storage was planted down after 3 days；Obtain no pollen lily former source kind after planting 2 years lowerly.
Wherein, the fertilizing management method after planting under step I and step N is as follows：
（1）After plantation, when stem-root grows 5-10mm, pour and apply 2 secondary root manures, fertilising for the first time is 12 days after plantation,
2nd fertilising is 22 days after plantation, and the component being applied fertilizer of taking root every time is as shown in the table, and that is, amount used in table is 1 time
Fertilising consumption；
Table 1
（2）At jointing stage, squaring period and the florescence growing in lily, applied a summer fertilizer every 7 days, apply altogether 10 times, each institute
The component applying summer fertilizer is as shown in the table, and that is, amount used in table is 1 fertilising consumption；
Table 2
Embodiment 3
A kind of breeding method of no pollen lily cultivar, comprises the steps：
Step A, July, in lily full-bloom stage, chooses male parent oriental hybrid lily flower pesticide grain, in not higher than 25 DEG C of warmhouse booth, right
Maternal oriental hybrid lily carries out artificial pollination, 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, daily observation, treats that capsule is ripe, but dorsal suture is not opened
Capsule is taken off, after twice of the alcohol wipe capsule being then 75% with mass concentration, capsule first being put into mass concentration is when splitting
Sterilize in 0.1% mercuric chloride solution 15min, places in the first mixed liquor sterilization 15min, uses aseptic water washing 3 times afterwards,
1min every time, obtains lily seed explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step C, strip off step on superclean bench（2）The lily seed explant obtaining, takes out and 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 2200 lx in intensity of illumination, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 13h/d, carry out germination induction；
First propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
pH 5.9；
Step D, after germination, each seed is peeled off kind of a skin, the every seed peeled off after kind of skin is individually forwarded to second
Propagating culture medium, is 2200 lx in intensity of illumination, and temperature is 25 ± 2 DEG C, under conditions of light application time is 13h/d, cultivates 35 days；
Second propagating culture medium of described 1L contains following component：
MS culture medium 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 the 3rd propagating culture medium after step D is cultivated 35 days, and continuing in intensity of illumination is 2200 lx, temperature
For 25 ± 2 DEG C, under conditions of light application time is 13h/d, cultivate 35 days；
3rd propagating culture medium of described 1L contains following component：
MS culture medium 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, the condition according to step E is further cultured for 35 days, it
The lily selecting all footpath 3.5cm of robust growth afterwards expands seed ball, is forwarded to root media, light culture 35 days；
The root media of described 1L contains following component：
MS culture medium 4.4g；
Methyl α-naphthyl acetate 0.9 mg/L；
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, the condition according to step F is further cultured for 35 days, obtains
To lily seed ball；It is 7 according to mass ratio:Turfy soil and water mixing, the mixture obtaining are packed by 3 to lily seed ball；Often
Grain lily seed ball uses 13g mixture；
Step H, packaged lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, room temperature storage 3 afterwards
Plant down after it；
Step I, it is seen that bulb is bloomed lowerly after planting 2 years, screening no pollen and the good lily of fancy points；
Step J, filters out no after pollen and the good lily of fancy points, after lily above-ground plant parts is withered, taking out should
Bulb, peels off outer scale, twice of the alcohol wipe being then 75% with mass concentration, and then first putting into mass concentration is 0.1%
Mercuric chloride solution in sterilize 15min, place in the first mixed liquor sterilization 15min, use aseptic water washing 3 times afterwards, every time
1min, obtains lily bud scale explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step K, step J lily bud scale explant is forwarded to the 4th propagating culture medium, is 2200 lx in intensity of illumination, temperature
For 25 ± 2 DEG C, under conditions of light application time is 13h/d, cultivate 35 days；
4th propagating culture medium of described 1L contains following component：
MS culture medium 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 continuing in intensity of illumination is 2200 lx, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 13h/d, cultivate 35 days；
5th propagating culture medium of described 1L contains following component：
MS culture medium 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 the 3rd propagating culture medium, and continuing in intensity of illumination is 2200 lx, temperature
Spend for 25 ± 2 DEG C, under conditions of light application time is 13h/d, cultivates 35 days, be forwarded to root media, change once within every 35 days
Culture medium, light culture, obtain no pollen lily seed ball；It is 7 according to mass ratio:Turfy soil and water are mixed by 3, the mixture obtaining
No pollen lily seed ball is packed；Every no pollen lily seed ball uses 13g mixture；
Step N, packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, afterwards
Room temperature storage was planted down after 3 days；Obtain no pollen lily former source kind after planting 2 years lowerly.
Wherein, the fertilizing management method after planting under step I and step N is as follows：
（1）After plantation, when stem-root grows 5-10mm, pour and apply 2 secondary root manures, fertilising for the first time is 10 days after plantation,
2nd fertilising is 20 days after plantation, and the component being applied fertilizer of taking root every time is as shown in the table, and that is, amount used in table is 1 time
Fertilising consumption；
Table 1
（2）At jointing stage, squaring period and the florescence growing in lily, applied a summer fertilizer every 7 days, apply altogether 10 times, each institute
The component applying summer fertilizer is as shown in the table, and that is, amount used in table is 1 fertilising consumption；
Table 2
The present embodiment obtained no pollen lily former source kind in 2015, as shown in Figure 1 and 2.
It is oriental hybrid lily Xi Bailun that the present embodiment is chosen maternal, and male parent is oriental hybrid lily Marco Polo.
The no pollen lily former source kind offspring's kind characteristic of the present embodiment gained：1. male sterility no pollen.2. Dan Zhihua
Head number 4-5.3. cut-flower production period 100-120 days.4. pattern：Lobe, outer lobe outer rim are pink, inner white.5. flower-shape：It is star-like,
Outer lobe outer rim big ripple.
General principle, principal character and the advantages of the present invention of the present invention have been shown and described above.The technology of the industry
, it should be appreciated that the present invention is not restricted to the described embodiments, the simply explanation described in above-described embodiment and specification is originally for personnel
The principle of invention, without departing from the spirit and scope of the present invention, the present invention also has various changes and modifications, these changes
Change and improvement both falls within scope of the claimed invention.Claimed scope by appending claims and its
Equivalent thereof.
Claims (2)
1. a kind of breeding method of no pollen lily cultivar is it is characterised in that comprise the steps：
Step A, July, in lily full-bloom stage, chooses male parent oriental hybrid lily flower pesticide grain, in not higher than 25 DEG C of warmhouse booth, right
Maternal oriental hybrid lily carries out artificial pollination, 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, daily observation, treats that capsule is ripe, but dorsal suture does not have
Capsule is taken off, after twice of the alcohol wipe capsule being then 75% with mass concentration, capsule first being put into mass concentration is during cracking
Sterilize in 0.1% mercuric chloride solution 15min, places in the first mixed liquor sterilization 15min, uses aseptic water washing 3 times afterwards,
1min every time, obtains lily seed explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step C, strip off step on superclean bench（2）The lily seed explant obtaining, takes out and 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 is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, carries out germination induction；
First propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8～6.0；
Step D, after germination, each seed is peeled off kind of a skin, and the every seed peeled off after kind of skin 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 bar of 12～14h/d
Under part, cultivate 30～40 days；
Second propagating culture medium of described 1L contains following component：
MS culture medium 4.4g；
Methyl α-naphthyl acetate 0.1mg/L；
6-benzyladenine 0.3～0.5mg/L；
Sucrose 30g/L；
Agarose 6g/L；
Deionized water surplus；
PH 5.8～6.0；
Step E, is forwarded to the 3rd propagating culture medium after step D is cultivated 30～40 days, continue intensity of illumination be 2000～
2500 lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days；
3rd propagating culture medium of described 1L contains following component：
MS culture medium 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.8～1.0mg/L
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, and the condition according to step E is further cultured for 30～
40 days, the lily selecting all footpath 3～4cm of robust growth afterwards expanded seed ball, was forwarded to root media, light culture 30～40 days；
The root media of described 1L contains following component：
MS culture medium 4.4g；
Methyl α-naphthyl acetate 0.8～1.0 mg/L；
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, the condition according to step F is further cultured for 30
～40 days, obtain lily seed ball；It is 7 according to mass ratio:Turfy soil and water mixing, the mixture obtaining are entered by 3 to lily seed ball
Row packaging；Every lily seed ball uses 10g～16.7g mixture；
Step H, packaged lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, room temperature storage 3 afterwards
Plant down after it；
Step I, it is seen that bulb is bloomed lowerly after planting 2 years, screening no pollen and the good lily of fancy points；
Step J, filters out no after pollen and the good lily of fancy points, after lily above-ground plant parts is withered, taking out should
Bulb, peels off outer scale, twice of the alcohol wipe being then 75% with mass concentration, and then first putting into mass concentration is 0.1%
Mercuric chloride solution in sterilize 15min, place in the first mixed liquor sterilization 15min, use aseptic water washing 3 times afterwards, every time
1min, obtains lily bud scale explant；
The preparation method of the first described mixed liquor is that in the liquor natrii hypochloritis that every 100ml mass concentration is 2%, dropping has 5
Tween-20, mixes, and obtains final product；
Step K, step J lily bud scale explant is forwarded to the 4th propagating culture medium, is 2000～2500 in intensity of illumination
Lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days；
4th propagating culture medium of described 1L contains following component：
MS culture medium 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.2～0.3mg/L
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 intensity of illumination be 2000～
2500 lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days；
5th propagating culture medium of described 1L contains following component：
MS culture medium 4.4g
Methyl α-naphthyl acetate 0.1mg/L
6-benzyladenine 0.5～0.8mg/L
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 the 3rd propagating culture medium, continue intensity of illumination be 2000～
2500 lx, temperature is 25 ± 2 DEG C, under conditions of light application time is 12～14h/d, cultivates 30～40 days, is forwarded to culture of rootage
Base, changes a subculture, light culture in every 30～40 days, obtains no pollen lily seed ball；It is 7 according to mass ratio:3 by turfy soil
Mix with water, the mixture obtaining is packed to no pollen lily seed ball；Every no pollen lily seed ball use 10g～
16.7g mixture；
Step N, packaged no pollen year lily seed ball is stored in -1 DEG C of freezer environment, outbound after storing 90 days, afterwards
Room temperature storage was planted down after 3 days；Obtain no pollen lily former source kind after planting 2 years lowerly.
2. the breeding method of no pollen lily cultivar according to claim 1 is it is characterised in that under step I and step N
Fertilizing management method after plantation is as follows：
（1）After plantation, when stem-root grows 5-10mm, pour and apply 2 secondary root manures, 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 being applied fertilizer of taking root every time is as shown in table 1；
Table 1
（2）At jointing stage, squaring period and the florescence growing in lily, applied a summer fertilizer every 7 days, apply altogether 10 times, each institute
The component applying summer fertilizer is as shown in table 2；
Table 2
.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| 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 |
| 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 |
| 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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| 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 |
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| 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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