CN108243944A - A kind of iris rapid breeding method and fine quality tissue culture propagation - Google Patents

Abstract

The invention discloses a kind of iris rapid breeding methods, it is cross-breeding using embryo culture technique, in particular with early stage embryo culture technique, overcome the incompatibility of distant hybridization, and the stringent control by including environmental condition and breeding technique to each link of entire breeding process, make that the new varieties cultivated plant strain growth is healthy and strong, character is excellent and shortens the period of breeding, so as to fulfill the rapid breeding of new varieties and selecting for improved seeds.The invention also discloses a kind of iris fine quality tissue culture propagations, for the new quality variety picked out, the advantages of maternal plant character can be kept in tissue culture procedures using bennet internode section, to grow directly from seeds, the bennet of improved Varieties that seedlings are stable, character of blooming is excellent, plant strain growth is healthy carries out tissue cultures as explant, and in different phase using particularly preferred culture medium, the iris cultivated under regular situation is made to emerge, and fast, stability is good, character is excellent and ornamental value is high.

Description

A kind of iris rapid breeding method and fine quality tissue culture propagation

Technical field

The present invention relates to iris culture raising technology fields, and in particular to a kind of iris rapid breeding method and high-quality Kind tissue culture propagation.

Background technology

Iris (Phalaenopsis) is orchid family Phalaenopsis perennial herb flowers, originates in Perenniporia martius Area, for flower-shape like butterfly, pattern is gorgeous, and the florescence is lasting, has high ornamental values and the economic values, is outside Now Domestic One of most fast-selling potted flower on flowers market.The pot variety of the marketization is that the plant type selected from cutting flower variety is compacter earliest Kind, the kinds for having a large amount of suitable pottings now bring out.Iris breeding method mainly has：Crossbreeding, mutation breeding With biotechnology breeding etc., wherein, crossbreeding be using generic cross introduce iris germplasm without character, be to formulate butterfly The important means of phalaenopsis novelty kind.And it is current, it needs just pick out new product with the time of 3 years by traditional breeding method Kind, and compared with can not ensure the hapanthous stability of seedling.

Phalaenopsis is seldom developed in single stem aerial orchid, lateral bud, and division propagation coefficient is extremely low, and main propagation method is adopted With aseptic seeding and by protocorm, callus etc., evoking adventive bud is bred again.But high-quality iris cenospecies utilizes sterile During seed propagation, offspring can detach, it is impossible to keep the merit of parent and the consistency of seedling, and use protocorm, Callus can increase the coefficient of variation of seedling when evoking adventive bud again breeds seedling.

Invention content

For more than technological deficiency, the technical problems to be solved by the invention are to provide a kind of iris rapid breeding method And fine quality tissue culture propagation, it is carried out by crossbreeding mode and to each link in entire breeding process stringent Control realizes the rapid breeding of new varieties iris, and the iris bennet segment of the fine quality to picking out is explant, Adventitious buds proliferation is carried out by inducing just training seedling, then just to train seedling, so as to fulfill the fast breeding of fine quality.

For solution more than technical problem, the technical solution adopted by the present invention is：

A kind of iris rapid breeding method, includes the following steps：

1）Hybridization pollination：It is parent by the butterfly orchid variety preferably gone out, using the maternal plant of robust growth when blooming pollinate miscellaneous It hands over, control temperature is at 27 ± 3 DEG C, and humid control is 60~80%；The seed of 120~150 days fruit maturations is used to broadcast after pollination Kind；

2）Aseptic seeding：The alcohol that mellow fruit mass concentration is 70~75% is impregnated 25~30 seconds, it is clear with sterile water After wash clean, sterilized 10~15 minutes with the mercuric chloride solution that mass concentration is 0.10~0.12%, then done with aseptic water washing Only, fruit is cut on superclean bench, by powdered seed uniform broadcasting in germination medium, sowing is sprouted after 30~38 days Hair forms protocorm；Germination medium used is：1/2MS+0.2~1mg/L6-BA+0.2~1mg/LNAA+20~30g/L sugarcanes The activated carbon of+5~8g/L of sugar+50~450mg/L of agar citric acids or/and pvp or/and vitamin C+0.15~0.20%+10~ 20% coconut juice, pH are 5.4~5.9, wherein, citric acid, polyvinylpyrrolidone pvp, vitamin C and activated carbon can be used as anti- Browning object mitigates browning in incubation；

3）Seedling culture：Protocorm is inoculated on Protocorm base, Protocorm base used is：3g/L spend precious No.1+ + 18~22% coconut juice+0.10 of+1.0~1.8% potato juice of 10~15g/L+5~8g/L of sucrose+3.5~5.5% bananas juices of agar ~0.18% activated carbon, pH are 5.3~5.5, wherein, it is sturdy that activated carbon can be conducive to plant strain growth；

Complete plantlet is grown into after 35~45 days, seedling culture medium is moved into, grows directly from seeds after continuing culture 35~45 days into stalwartness Seedling；Seedling growth medium used is：3g/L spends precious No.1+0.1~1mg/L6-BA+0.4~2mg/LNAA+10~15g/L Sucrose+5~+0.10~0.18% activated carbon of+1.0~1.5% potato juice of+7~9% coconut juice of 8g/L agar, pH are 5.1~5.6；

4）Bottle outlet is cultivated：Test tube seedling length when having 3~4 leaves, opens bottle stopper, at room temperature hardening one week, i.e., to 2~3cm high It is planted outside removable bottle outlet；During bottle outlet, the culture medium of root is cleaned, wind after 4~6min is impregnated in 0.1% liquor potassic permanganate Moisture on cured leaf face, is planted in mixed-matrix, keeps ventilation and heat preservation, moisturizing, 5000~8000Lux of illumination;It is used go out The mixed-matrix of bottle transplanting includes the raw material using following weight proportion：Fertile soil：Wood fragments charcoal：Sphagna：Volcanic rock foundation stone is 1.5 ~2.5:0.9~1.1:0.8~1.2:1；

5）Seedling stage plants：After 30~45 days, every 5~10 days roots fertilising is primary, each 25~35ppm of nitrogen, phosphorus, potassium concn, seaweed Smart 5500 times of liquid, in 8000~10000Lux of illumination, 25 ± 1 DEG C of temperature keeps continuing culture 90~120 days under ventilation condition;

Every 5~10 days roots fertilising is primary later, each 30~40ppm of nitrogen, phosphorus, potassium concn, 5000 times of liquid of algae essence, in illumination 11000~15000Lux, 26 ± 2 DEG C of temperature are cultivated 120~150 days under the conditions of relative humidity 60~85%；

Every 5~10 days roots fertilising is primary later, and nitrogen, phosphorus, potassium concn are respectively 40~50ppm, 4500 times of liquid of algae essence, in illumination 17000~20000Lux, 28 ± 2 DEG C of temperature are planted 120~150 days under the conditions of relative humidity 60~80%, seedling plantation into It is ripe；

Wherein, algae essence same, the Ji Zhiwuying that contains various indispensable elements needed for plant growth, amino acid and plant growth substance Substance, bioactive substance, the plant stress-resistance factor are supported in one, can be met needed for seedling stage plant strain growth；

6）The flower forcing of seedling：Move into progress flower forcing processing in greenhouse after seedling plantation is ripe, every 5~10 days roots apply nitrogen, Phosphorus, potassium balance fertilizer are primary, fertilizer EC values 1.0~1.2,10000~12000Lux of illumination, 30 ± 2 DEG C of day temperature, nocturnal temperature 25±1℃；After January, the high potassium flower forcing fertilizer of high phosphorus is applied within every 5~7 days once, fertilizer EC values 1.0~1.2, illumination 12000~ 15000Lux, 25 ± 1 DEG C of day temperature, 19 ± 1 DEG C of nocturnal temperature；After 75~90 days, nitrogen, phosphorus, potassium balance fertilizer are applied within every 5~7 days Once, fertilizer EC values 0.8~1.0,15000~20000Lux of illumination, 26 ± 2 DEG C of day temperature, 20 ± 2 DEG C of nocturnal temperature until It blooms；

7）Pick out the iris new varieties that robust growth, quality are good, fancy points is excellent.

The invention also discloses carry out iris fine quality tissue after cultivating new varieties iris using the above method Propagation method is cultivated, is included the following steps：

1）Explant is disinfected：Using the bennet of new varieties iris picked out as explant, removed with the blade sterilized The outer bract of axillary bud on bennet section position, and surface is dabbed with the alcohol that mass concentration is 70~75%, it is clean with sterile water wash Afterwards, bennet is cut into every section of bennet segment for including a section position, is put into the chlorination that mass concentration is 0.10~0.12% It soaking disinfection 8~10 minutes and is constantly shaken, then bennet segment is taken out in mercury solution, it is clean with sterile water wash；

2）The just sprouting of training seedling：The explant both ends blade sterilized after disinfection is cut, is inoculated into germination medium, 23~26 DEG C of temperature, preceding 10 days light cultures, culture to 22~25 days, makes explant under the conditions of 2000~2500Lux of light intensity later Body sprouts to obtain just training seedling；Germination medium used is：1/2MS+1~3mg/L6-BA+0.5~1.5mg/LTDZ+0.1~ 0.3mg/LNAA+0.2~0.3mg/L vitamin C+0.2~0.3g/L activated carbon+20~30g/L+6~7g/L of sucrose agar+ 0.01~0.05mg/LDA-6, pH be 5.4~5.8, wherein, plant growth regulator TDZ can promote plant sprout regeneration and The suspend mode of bud is broken in breeding, and diethyl aminoethyl hexanoate DA-6 can promote the fast-germination of explant, improve reproduction speed；

3）Just training seedling proliferation culture：First training seedling is inoculated in proliferated culture medium, 23~26 DEG C, illumination 12h/d of temperature, light intensity It is cultivated 40~45 days under the conditions of 1500~2000Lux, obtains Multiple Buds；Proliferated culture medium used is：MS+1.0~3.5mg/L6- Coconut juice+0.2~0.3g/L activated carbon+20~30g/L sugarcanes of BA+0.2~1.0mg/LNAA+0.1~0.3mg/LTDZ+12~20% Sugar+6~7g/L agar+12~18g/L corn oligopeptide powders, pH are 5.4~5.6, wherein, corn oligopeptide powder can enhance something lost The stability of transmissibility shape；

4）Adventitious buds proliferation：The Multiple Buds of formation are subjected to shoot proliferation on new proliferated culture medium, in 23~25 DEG C of temperature, It is cultivated 15~20 days under the conditions of light application time 12h/d, 2000~2500Lux of light intensity;

5）Strengthening seedling and rooting culture：After proliferation and subculture culture, the complete seedlings of high more than 2cm is selected to be transferred in strong seedling culture base, Cultivated 15~20 days under the conditions of 23~25 DEG C, light application time 12h/d, 2000~2500Lux of light intensity of temperature, backward strong seedling culture 0.18~0.32% activated carbon is added in base, in 27~29 DEG C, light application time 15h/d of temperature, light intensity 2500~3000Lux conditions Under continue to cultivate；Strong seedling culture base used is：3g/L spends precious No.1+2.5~3.5mg/L6-BA+0.4~0.6mg/L NAA+20 + 8~12%+0.01~0.05mg/LDA-6 of coconut juice of~27g/L+5~9g/L of sucrose agar, activated carbon is added during this can be with It is sturdy and prevent brown stain to be effectively conducive to plant strain growth；

6）Transplanting：When test tube seedling grows to 4~5cm high, root system is up to 2 or more and healthy and strong vibrant, during 1~3, leaf, by test tube Seedling is placed on natural light lower refining seedling 7~12 days, opens bottle cap hardening 1~2 day；Cultivating seedling is taken out, is cleaned with clear water and is attached to root Culture medium, then transplanting to include wood fragments charcoal, thick coconut husk, fertile soil and the mixed-matrix for the sphagna impregnated in, keep phase To humidity 70~85%, 20~25 DEG C of temperature.

By adopting the above-described technical solution, the beneficial effects of the invention are as follows：

Due to a kind of iris rapid breeding method provided by the invention, carry out seed using the ripe fruit pod after hybridization pollination and lure Protocorm, and the stringent control by including environmental condition and breeding technique to each link of entire breeding process are led, makes to cultivate New varieties plant strain growth is healthy and strong, character is excellent, so as to fulfill the rapid breeding of new varieties and selecting for improved seeds, entirely educate Kind process substantially reduced the period of breeding of new variety between 26~28 months.

Due to a kind of iris fine quality tissue culture propagation provided by the invention, using bennet internode section in group The advantages of maternal plant character can be kept in incubation is knitted, seedlings are stable, character of blooming is excellent, plant strain growth is healthy to grow directly from seeds The bennets of improved Varieties carry out tissue cultures, and make using particularly preferred culture medium in different phase as explant Survival rate and inductivity are up to 90%, compared with prior art, with emergence is fast, planting percent is high, stability is good, character is excellent The advantages of.

In conclusion the present invention is cross-breeding using embryo culture technique, in particular with early stage embryo culture technique, gram The incompatibility of distant hybridization has been taken, and has shortened the period of breeding, while for the new quality variety picked out, explant is adopted The bennet that can keep the inhereditary feature of maternal stabilization with offspring carries out tissue cultures breeding, and making in normal conditions can be acquired The combination that shape is stable and ornamental value is high.

Specific embodiment

With reference to specific embodiment, the present invention is further explained.

One iris rapid breeding method of embodiment

In the present embodiment, the rapid breeding of iris new varieties is carried out using the method for crossbreeding, is as follows：

1）Hybridization pollination：It is parent by the butterfly orchid variety preferably gone out, using the maternal plant of robust growth when blooming pollinate miscellaneous It hands over, control temperature is at 27 ± 3 DEG C, and humid control is 60~80%；The seed of 120~150 days fruit maturations is used to broadcast after pollination Kind；

2）Aseptic seeding：The alcohol that mellow fruit mass concentration is 70~75% is impregnated 25~30 seconds, it is clear with sterile water After wash clean, sterilized 10~15 minutes with the mercuric chloride solution that mass concentration is 0.10~0.12%, then done with aseptic water washing Only, fruit is cut on superclean bench, by powdered seed uniform broadcasting in germination medium, sowing is sprouted after 30~38 days Hair forms protocorm；Germination medium used is：1/2MS+0.2~1mg/L6-BA+0.2~1mg/LNAA+20~30g/L sugarcanes The activated carbon of+5~8g/L of sugar+50~450mg/L of agar citric acids or/and pvp or/and vitamin C+0.15~0.20%+10~ 20% coconut juice, pH are 5.4~5.9, wherein, citric acid, polyvinylpyrrolidone pvp, vitamin C and activated carbon can be used as anti- Browning object mitigates browning in incubation；

3）Seedling culture：Protocorm is inoculated on Protocorm base, Protocorm base used is：3g/L spend precious No.1+ + 18~22% coconut juice+0.10 of+1.0~1.8% potato juice of 10~15g/L+5~8g/L of sucrose+3.5~5.5% bananas juices of agar ~0.18% activated carbon, pH are 5.3~5.5；

Complete plantlet is grown into after 35~45 days, seedling culture medium is moved into, grows directly from seeds after continuing culture 35~45 days into stalwartness Seedling；Seedling growth medium used is：3g/L spends precious No.1+0.1~1mg/L6-BA+0.4~2mg/LNAA+10~15g/L Sucrose+5~+0.10~0.18% activated carbon of+1.0~1.5% potato juice of+7~9% coconut juice of 8g/L agar, pH are 5.1~5.6；

Wherein, it is sturdy can be conducive to plant strain growth for activated carbon；

4）Bottle outlet is cultivated：Test tube seedling length when having 3~4 leaves, opens bottle stopper, at room temperature hardening one week, i.e., to 2~3cm high It is planted outside removable bottle outlet；During bottle outlet, the culture medium of root is cleaned, wind after 4~6min is impregnated in 0.1% liquor potassic permanganate Moisture on cured leaf face, is planted in mixed-matrix, keeps ventilation and heat preservation, moisturizing, 5000~8000Lux of illumination;It is used go out The mixed-matrix of bottle transplanting includes the raw material using following weight proportion：Fertile soil：Wood fragments charcoal：Sphagna：Volcanic rock foundation stone is 1.5 ~2.5:0.9~1.1:0.8~1.2:1；

5）Seedling stage plants：After 30~45 days, every 5~10 days roots fertilising is primary, each 25~35ppm of nitrogen, phosphorus, potassium concn, seaweed Smart 5500 times of liquid, in 8000~10000Lux of illumination, 25 ± 1 DEG C of temperature keeps continuing culture 90~120 days under ventilation condition;

Every 5~10 days roots fertilising is primary later, each 30~40ppm of nitrogen, phosphorus, potassium concn, 5000 times of liquid of algae essence, in illumination 11000~15000Lux, 26 ± 2 DEG C of temperature are cultivated 120~150 days under the conditions of relative humidity 60~85%；

Every 5~10 days roots fertilising is primary later, and nitrogen, phosphorus, potassium concn are respectively 40~50ppm, 4500 times of liquid of algae essence, in illumination 17000~20000Lux, 28 ± 2 DEG C of temperature are planted 120~150 days under the conditions of relative humidity 60~80%, seedling plantation into It is ripe；

Wherein, algae essence same, the Ji Zhiwuying that contains various indispensable elements needed for plant growth, amino acid and plant growth substance Substance, bioactive substance, the plant stress-resistance factor are supported in one, can be met needed for seedling stage plant strain growth；

6）The flower forcing of seedling：Move into progress flower forcing processing in greenhouse after seedling plantation is ripe, every 5~10 days roots apply nitrogen, Phosphorus, potassium balance fertilizer are primary, fertilizer EC values 1.0~1.2,10000~12000Lux of illumination, 30 ± 2 DEG C of day temperature, nocturnal temperature 25±1℃；After January, the high potassium flower forcing fertilizer of high phosphorus is applied within every 5~7 days once, fertilizer EC values 1.0~1.2, illumination 12000~ 15000Lux, 25 ± 1 DEG C of day temperature, 19 ± 1 DEG C of nocturnal temperature；After 75~90 days, nitrogen, phosphorus, potassium balance fertilizer are applied within every 5~7 days Once, fertilizer EC values 0.8~1.0,15000~20000Lux of illumination, 26 ± 2 DEG C of day temperature, 20 ± 2 DEG C of nocturnal temperature until It blooms；

7）Pick out the iris new varieties that robust growth, quality are good, fancy points is excellent.

The present embodiment uses three groups of experiments, and each group is two groups of contrast experiments of experimental group and control group respectively, and each group compares Parallel laboratory test three times is done in experiment, and the male parent of hybridization used and the number difference of female parent are as shown in the table：

Wherein, three groups of experimental groups carry out breeding using rapid breeding method described above, and control group uses traditional breeding way, Terminate to the flower forcing phase, when blooming, breeding experiment periods terminate, as shown in the table the time required to the breeding all stage：

As can be seen from the above table, using a kind of iris rapid breeding method provided by the invention, using after hybridization pollination into Ripe fruit pod carries out Seed inducement protocorm, and by including the tight of environmental condition and breeding technique to each link of entire breeding process Lattice control, and make the new varieties plant strain growth cultivated stalwartness, character excellent, rapid breeding and excellent product so as to fulfill new varieties Kind is selected, and entire breeding process substantially reduced the period of breeding of new variety between 26~28 months.

Two iris fine quality tissue culture propagation of embodiment

In the present embodiment, after cultivating new varieties iris using one the method for embodiment, the reality of contrast experiment's group three is selected Test group 3, i.e. the butterfly orchid variety that male parent number YT-0231, maternal number YT-0215 are bred as carries out culture propagation method, including Following steps：

1）Explant is disinfected：Using the bennet of new varieties iris picked out as explant, removed with the blade sterilized The outer bract of axillary bud on bennet section position, and surface is dabbed with the alcohol that mass concentration is 70~75%, it is clean with sterile water wash Afterwards, bennet is cut into every section of bennet segment for including a section position, is put into the chlorination that mass concentration is 0.10~0.12% It soaking disinfection 8~10 minutes and is constantly shaken, then bennet segment is taken out in mercury solution, it is clean with sterile water wash；

2）The just sprouting of training seedling：The explant both ends blade sterilized after disinfection is cut, is inoculated into germination medium, 23~26 DEG C of temperature, preceding 10 days light cultures, culture to 22~25 days, makes explant under the conditions of 2000~2500Lux of light intensity later Body sprouts to obtain just training seedling；Germination medium used is：1/2MS+1~3mg/L6-BA+0.5~1.5mg/LTDZ+0.1~ 0.3mg/LNAA+0.2~0.3mg/L vitamin C+0.2~0.3g/L activated carbon+20~30g/L+6~7g/L of sucrose agar+ 0.01~0.05mg/LDA-6, pH are 5.4~5.8, wherein, diethyl aminoethyl hexanoate DA-6 can promote the fast-germination of explant, improve Reproduction speed；

3）Just training seedling proliferation culture：First training seedling is inoculated in proliferated culture medium, 23~26 DEG C, illumination 12h/d of temperature, light intensity It is cultivated 40~45 days under the conditions of 1500~2000Lux, obtains Multiple Buds；Proliferated culture medium used is：MS+1.0~3.5mg/L6- Coconut juice+0.2~0.3g/L activated carbon+20~30g/L sugarcanes of BA+0.2~1.0mg/LNAA+0.1~0.3mg/LTDZ+12~20% Sugar+6~7g/L agar+12~18g/L corn oligopeptide powders, pH are 5.4~5.6, wherein, plant growth regulator TDZ can be with Promote the regeneration and breeding of plant sprout, break the suspend mode of bud, corn oligopeptide powder can enhance the stability of inhereditary feature；

4）Adventitious buds proliferation：The Multiple Buds of formation are subjected to shoot proliferation on new proliferated culture medium, in 23~25 DEG C of temperature, It is cultivated 15~20 days under the conditions of light application time 12h/d, 2000~2500Lux of light intensity;

5）Strengthening seedling and rooting culture：After proliferation and subculture culture, the complete seedlings of high more than 2cm is selected to be transferred in strong seedling culture base, Cultivated 15~20 days under the conditions of 23~25 DEG C, light application time 12h/d, 2000~2500Lux of light intensity of temperature, backward strong seedling culture 0.18~0.32% activated carbon is added in base, in 27~29 DEG C, light application time 15h/d of temperature, light intensity 2500~3000Lux conditions Under continue to cultivate；Strong seedling culture base used is：3g/L spends precious No.1+2.5~3.5mg/L6-BA+0.4~0.6mg/L NAA+20 + 8~12%+0.01~0.05mg/LDA-6 of coconut juice of~27g/L+5~9g/L of sucrose agar；

6）Transplanting：When test tube seedling grows to 4~5cm high, root system is up to 2 or more and healthy and strong vibrant, during 1~3, leaf, by test tube Seedling is placed on natural light lower refining seedling 7~12 days, opens bottle cap hardening 1~2 day；Cultivating seedling is taken out, is cleaned with clear water and is attached to root Culture medium, then transplanting to include wood fragments charcoal, thick coconut husk, fertile soil and the mixed-matrix for the sphagna impregnated in, keep phase To humidity 70~85%, 20~25 DEG C of temperature.

The present embodiment is simultaneously using the different tissues of experimental group 3 as explant, using identical tissue culture propagation Tissue cultures are carried out, i.e., respectively using seed, bennet, blade, stem apex, the tip of a root as explant, each group does experimental group and right respectively According to two groups of contrast experiments of group, wherein, explant used in experimental group is bennet, and each group contrast experiment does parallel laboratory test three times.

In this control experiment, culture medium combination 1 is according to ingredient used in the present embodiment and each culture medium of proportioning Combination, culture medium combination 2 is does not add corn oligopeptide powder, diethyl aminoethyl hexanoate DA-6 and plant in corresponding culture medium in culture medium combination 1 Object growth regulator TDZ.

Wherein, inhereditary feature grading in, to maternal character heredity grade 1~5 grade gradually enhance, 1 grade be trait segregation, 5 Grade is retains maternal character；1~5 grade of grading is influenced on maternal plant to incrementally increase, 1 grade is not influence maternal plant, and 5 grades are to influence maternal plant Greatly.Experimental comparison results are as shown in the table：

It can be seen from embodiment data compared with each control group, using bennet as explant, by the tissue cultures of the present invention Propagation method, the iris new talent of culture have higher induction germination rate, and new talent is more healthy and strong, and survival rate is high after hardening.Together When, increase foster germination rate, new shoot survival percent of new talent etc. there are bigger difference with different culture media combination between control group is organized, Each performance indicator of new talent of 1 culture of culture medium combination used in the present invention is more excellent, shows the method for the present invention and culture medium Superiority.

In conclusion the present invention is cross-breeding using embryo culture technique, in particular with early stage embryo culture technique, gram The incompatibility of distant hybridization has been taken, and has shortened the period of breeding, while for the new quality variety picked out, explant is adopted The bennet that can keep the inhereditary feature of maternal stabilization with offspring carries out tissue cultures breeding, and making in normal conditions can be acquired The combination that shape is stable and ornamental value is high.

The above-described embodiments are merely illustrative of preferred embodiments of the present invention, not to the structure of the present invention Think and range is defined.Therefore, under the premise of present invention setting design is not departed from, ordinary people in the field is to the present invention's The all variations and modifications that technical solution is made should all drop into protection scope of the present invention, the claimed technology of the present invention Content has all been recorded in detail in the claims.

Claims (10)

1. A kind of 1. iris rapid breeding method, it is characterised in that：Include the following steps：

   1）Hybridization pollination：It is parent by the butterfly orchid variety preferably gone out, using the maternal plant of robust growth when blooming pollinate miscellaneous It hands over, control temperature is at 27 ± 3 DEG C, and humid control is 60~80%；The seed of 120~150 days fruit maturations is used to broadcast after pollination Kind；

   2）Aseptic seeding：The alcohol that mellow fruit mass concentration is 70~75% is impregnated 25~30 seconds, it is clear with sterile water After wash clean, sterilized 10~15 minutes with the mercuric chloride solution that mass concentration is 0.10~0.12%, then done with aseptic water washing Only, fruit is cut on superclean bench, by powdered seed uniform broadcasting in germination medium, sowing is sprouted after 30~38 days Hair forms protocorm；

   3）Seedling culture：Protocorm is inoculated on Protocorm base, complete plantlet is grown into after 35~45 days, is moved into Seedling culture medium continues after culture 35~45 days into healthy and strong seedling；

   4）Bottle outlet is cultivated：Test tube seedling length when having 3~4 leaves, opens bottle stopper, at room temperature hardening one week, i.e., to 2~3cm high It is planted outside removable bottle outlet；During bottle outlet, the culture medium of root is cleaned, wind after 4~6min is impregnated in 0.1% liquor potassic permanganate Moisture on cured leaf face, is planted in mixed-matrix, keeps ventilation and heat preservation, moisturizing, 5000~8000Lux of illumination;

   5）Seedling stage plants：After 30~45 days, every 5~10 days roots fertilising is primary, each 25~35ppm of nitrogen, phosphorus, potassium concn, seaweed Smart 5500 times of liquid, in 8000~10000Lux of illumination, 25 ± 1 DEG C of temperature keeps continuing culture 90~120 days under ventilation condition;

   Every 5~10 days roots fertilising is primary later, each 30~40ppm of nitrogen, phosphorus, potassium concn, 5000 times of liquid of algae essence, in illumination 11000~15000Lux, 26 ± 2 DEG C of temperature are cultivated 120~150 days under the conditions of relative humidity 60~85%；

   Every 5~10 days roots fertilising is primary later, and nitrogen, phosphorus, potassium concn are respectively 40~50ppm, 4500 times of liquid of algae essence, in illumination 17000~20000Lux, 28 ± 2 DEG C of temperature are planted 120~150 days under the conditions of relative humidity 60~80%, seedling plantation into It is ripe；

   6）The flower forcing of seedling：Move into progress flower forcing processing in greenhouse after seedling plantation is ripe, every 5~10 days roots apply nitrogen, Phosphorus, potassium balance fertilizer are primary, fertilizer EC values 1.0~1.2,10000~12000Lux of illumination, 30 ± 2 DEG C of day temperature, nocturnal temperature 25±1℃；After January, the high potassium flower forcing fertilizer of high phosphorus is applied within every 5~7 days once, fertilizer EC values 1.0~1.2, illumination 12000~ 15000Lux, 25 ± 1 DEG C of day temperature, 19 ± 1 DEG C of nocturnal temperature；After 75~90 days, nitrogen, phosphorus, potassium balance fertilizer are applied within every 5~7 days Once, fertilizer EC values 0.8~1.0,15000~20000Lux of illumination, 26 ± 2 DEG C of day temperature, 20 ± 2 DEG C of nocturnal temperature until It blooms；

   7）Pick out the iris new varieties that robust growth, quality are good, fancy points is excellent.
2. 2. a kind of iris rapid breeding method according to claim 1, it is characterised in that：Step 2）It is used to sprout culture Base is：1/2MS+0.2~1mg/L6-BA+0.2~+50~450mg/L of 1mg/LNAA+20~30g/L sucrose+5~8g/L agar Citric acid or/and pvp or/and vitamin C+0.15~+10~20% coconut juice of 0.20% activated carbon, pH are 5.4~5.9.
3. 3. a kind of iris rapid breeding method according to claim 1, it is characterised in that：Step 3）Protocorm training used Foster base is：3g/L spends precious No.1+10~+1.0~1.8% potato of+3.5~5.5% bananas juice of 15g/L+5~8g/L of sucrose agar Juice+18~+0.10~0.18% activated carbon of 22% coconut juice, pH are 5.3~5.5.
4. 4. a kind of iris rapid breeding method according to claim 1, it is characterised in that：Step 3）Step 4）It is used into Seedling growth medium is：3g/L spend precious No.1+0.1~1mg/L6-BA+0.4~2mg/LNAA+10~15g/L sucrose+5~ + 0.10~0.18% activated carbon of+1.0~1.5% potato juice of+7~9% coconut juice of 8g/L agar, pH are 5.1~5.6.
5. 5. a kind of iris rapid breeding method according to claim 1, it is characterised in that：The mixing of bottle outlet transplanting used Matrix includes the raw material using following weight proportion：Fertile soil：Wood fragments charcoal：Sphagna：Volcanic rock foundation stone is 1.5~2.5:0.9~ 1.1:0.8~1.2:1.
6. 6. according to a kind of iris fine quality tissue culture propagation of claim 1-5 any one of them, feature exists In：Include the following steps：

   1）Explant is disinfected：Using the bennet of new varieties iris picked out as explant, removed with the blade sterilized The outer bract of axillary bud on bennet section position, and surface is dabbed with the alcohol that mass concentration is 70~75%, it is clean with sterile water wash Afterwards, bennet is cut into every section of bennet segment for including a section position, is put into the chlorination that mass concentration is 0.10~0.12% It soaking disinfection 8~10 minutes and is constantly shaken, then bennet segment is taken out in mercury solution, it is clean with sterile water wash；

   2）The just sprouting of training seedling：The explant both ends blade sterilized after disinfection is cut, is inoculated into germination medium, 23~26 DEG C of temperature, preceding 10 days light cultures, culture to 22~25 days, makes explant under the conditions of 2000~2500Lux of light intensity later Body sprouts to obtain just training seedling；

   3）Just training seedling proliferation culture：First training seedling is inoculated in proliferated culture medium, 23~26 DEG C, illumination 12h/d of temperature, light intensity It is cultivated 40~45 days under the conditions of 1500~2000Lux, obtains Multiple Buds；

   4）Adventitious buds proliferation：The Multiple Buds of formation are subjected to shoot proliferation on new proliferated culture medium, in 23~25 DEG C of temperature, It is cultivated 15~20 days under the conditions of light application time 12h/d, 2000~2500Lux of light intensity;

   5）Strengthening seedling and rooting culture：After proliferation and subculture culture, the complete seedlings of high more than 2cm is selected to be transferred in strong seedling culture base, Cultivated 15~20 days under the conditions of 23~25 DEG C, light application time 12h/d, 2000~2500Lux of light intensity of temperature, backward strong seedling culture 0.18~0.32% activated carbon is added in base, in 27~29 DEG C, light application time 15h/d of temperature, light intensity 2500~3000Lux conditions Under continue to cultivate；

   6）Transplanting：When test tube seedling grows to 4~5cm high, root system is up to 2 or more and healthy and strong vibrant, during 1~3, leaf, by test tube Seedling is placed on natural light lower refining seedling 7~12 days, opens bottle cap hardening 1~2 day；Cultivating seedling is taken out, is cleaned with clear water and is attached to root Culture medium, then transplant into mixed-matrix, keep relative humidity 70~85%, 20~25 DEG C of temperature.
7. 7. a kind of iris fine quality tissue culture propagation according to claim 6, it is characterised in that：Step 2） Germination medium used is：1/2MS+1~3mg/L6-BA+0.5~1.5mg/LTDZ+0.1~0.3mg/LNAA+0.2~ + 0.01~0.05mg/LDA- of 0.3mg/L vitamin C+0.2~0.3g/L activated carbon+20~30g/L sucrose+6~7g/L agar 6, pH be 5.4~5.8.
8. 8. a kind of iris fine quality tissue culture propagation according to claim 6, it is characterised in that：Step 3） Proliferated culture medium used is：MS+1.0~3.5mg/L6-BA+0.2~1.0mg/LNAA+0.1~0.3mg/LTDZ+12~20% Coconut juice+0.2~0.3g/L activated carbon+20~30g/L sucrose+6~7g/L agar+12~18g/L corn oligopeptide powders, pH are 5.4~5.6.
9. 9. a kind of iris fine quality tissue culture propagation according to claim 6, it is characterised in that：Step 5） Strong seedling culture base used is：3g/L spends precious No.1+2.5~3.5mg/L6-BA+0.4~0.6mg/L NAA+20~27g/L sucrose + 8~12%+0.01~0.05mg/LDA-6 of coconut juice of+5~9g/L agar.
10. 10. a kind of iris fine quality tissue culture propagation according to claim 6, it is characterised in that：Step 6）Mixed-matrix used includes wood fragments charcoal, thick coconut husk, fertile soil and the sphagna impregnated.