CN107864854A - Floral leaf Wu's wind grass fast breeding method based on tissue cultures - Google Patents
Floral leaf Wu's wind grass fast breeding method based on tissue cultures Download PDFInfo
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- CN107864854A CN107864854A CN201610854285.4A CN201610854285A CN107864854A CN 107864854 A CN107864854 A CN 107864854A CN 201610854285 A CN201610854285 A CN 201610854285A CN 107864854 A CN107864854 A CN 107864854A
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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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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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Abstract
The present invention relates to a kind of floral leaf Wu's wind grass fast breeding method based on tissue cultures, comprise the following steps：(1) bud inducement cultivation base, adventitious bud proliferation culture medium, strong seedling culture base, root media, the tissue cultures of (2) floral leaf Wu wind grass are prepared.Compared with prior art, the present invention is by tissue culture technique, year breeding coefficient bring up to 1:50000, cost lowers 0.8 1 yuan/plant, because the floral leaf of blade is yellowish green chimera, tissue culture difficulty is larger, and by adjusting the ratio of auxin and the basic element of cell division, leaf characteristic is obvious, the regularity of reproduction speed and seedling is greatly improved, preferably keeps original maternal character.
Description
Technical field
It is quick more particularly, to a kind of floral leaf Wu wind grass based on tissue cultures the present invention relates to a kind of plant breeding methods
Mating system.
Background technology
Floral leaf Farfugium japonicum is composite family Farfugium japonicum platymiscium, perennial evergreen draft, basal leaf rosette-stape, kidney shape, leaf
It is greenish-yellow chimeric.The 10-11 months bloom, yellow.The cultivation of floral leaf Farfugium japonicum is easy and evergreen, and greenery Cheng Cong is extremely attractive in appearance.In gardens
More group planting are transplanted, under the woods in flower border, roadside.Floral leaf Farfugium japonicum resistance to half is shady, can do sparse woods cover plant again, be good
Presbyopic glasses and garden material.The Introduced Cultivars negligible amounts introduced as foreign countries, its plant division is slow, the huge market demand, seedling supply
It is restricted.
Chinese patent CN103734019A discloses a kind of method for tissue culture of New Zealand flax, and this method bag is prepared different
The culture medium in stage carries out tissue cultures, can improve the regularity of reproduction speed and seedling, preferably keeps original female parent
The advantages that character.But because New Zealand flax and floral leaf Wu wind grass habit and required environment difference of breeding are larger, above-mentioned tissue
Cultural method cannot be directly used to the reproductive process of floral leaf Wu wind grass.
The content of the invention
It is an object of the present invention to overcome the above-mentioned drawbacks of the prior art and provide one kind breed speed it is fast, into
The high floral leaf Wu's wind grass fast breeding method based on tissue cultures of motility rate.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of floral leaf Wu's wind grass fast breeding method based on tissue cultures, comprises the following steps：
(1) preparation of culture medium, the component and content of each stage culture medium of tissue culture are included：
1.1st, bud inducement cultivation base：MS+BA 1-3mg/L+NAA 0.1-0.3mg/L；
1.2nd, adventitious bud proliferation culture medium：MS+6-BA 0.5-1.5mg/L+NAA 0.05-0.15mg/L；
1.3rd, strong seedling culture base：MS+6-BA 0.1-0.3mg/L+NAA 0.01-0.03mg/L；
1.4th, root media：MS+NAA 0.05-0.15mg/L；
The pH5.8 of above-mentioned various culture mediums, sucrose 30g/L, agar 6g/L are added, cultivation temperature is 25 ± 1 DEG C, and illumination is
1100-1300Lx；
(2) tissue cultures of floral leaf Wu wind grass：
2.1st, the acquisition of sterilizable material
Floral leaf Wu's wind grass takes the budlet of aerial part in the month in spring 3-4, and 1h is rinsed after superclean bench with running water
On, 30s is soaked with 75% ethanol, 1 ‰ mercuric chloride soak 15min, aseptic water washing 5-6 times, and aseptic filter paper blots surface moisture content,
Take budlet base portion to be cut into 1cm length, be inoculated on bud inducement cultivation base；
2.2nd, the differentiation of bud and propagation
Budlet is inoculated on inducing culture after 30 days, and budlet base portion starts to expand, and yellow green projection is occurred, is further cultured for 25
Its visible obvious callus, continue culture 30 days, cut to be put on adventitious bud proliferation culture medium with bud callus and cultivate；
2.3rd, adventitious bud strong seedling culture
On adventitious bud proliferation culture medium, the only 3-4 strains of Multiple Buds every clump induced can extend, and remaining, which is in, downgrades
State, after being divided into little Cong or simple bud, strong seedling culture base is put into, adventitious bud is extended rapidly, and 2-3cm can be grown to after 20 days；
2.4th, culture of rootage
1.5-2cm plantlet is taken, root induction in root media of transferring, seedling base portion differentiates many after 20 days
The root restriction of white, can grow to 1-2cm after 30 days；
2.5th, hardening and transplanting
After culture of rootage 28-32 days, when root system length is to 1-2cm, the aseptic seedling of well developed root system robust growth is selected, interior is opened
Bottle hardening 3 days, cleans root agar after taking-up, hardening is tamed in greenhouse after 40 days, i.e. transplanting is outdoor, gives rich water quality management,
Transplanting survival rate is more than 90%.
Preferably, the component and content of described bud inducement cultivation base are：MS+BA 2mg/L+NAA 0.2mg/L, should
Budlet growing way is best under culture medium.
Preferably, the component and content of described adventitious bud proliferation culture medium are：MS+6-BA1mg/L+NAA 0.1mg/
L, under the culture medium, base portion callus is relatively more, but does not influence the propagation of adventitious bud, and adventitious bud growth is rapid, no chrysanthemum or
Become the bad phenomenon such as green entirely, grown in the culture good.
Preferably, the component and content of described strong seedling culture base are：MS+6-BA 0.2mg/L+NAA0.02mg/L.
Preferably, the component and content of described root media are：MS+NAA0.1mg/L, under the culture medium, root system
Sturdy, fibrous root is numerous, rooting rate 100%.
It is very big especially for the requirement difference of culture medium because different plants require different for condition of culture, wherein
The change of the culture medium main component or content of one link is possible to cause to breed substantially reducing for success rate.Therefore, originally
Invention passes through permanent experimentation, and according to the endemic plant growth chracteristic and growth demand of floral leaf Wu wind grass, have studied suitable
A variety of culture mediums of floral leaf Wu's wind grass fast breeding are closed, on the medium base using these special developments, with reference to the present invention
Condition of culture, successful incubation floral leaf Wu wind grass.
The present invention by tissue culture technique, year breeding coefficient bring up to 1:50000, cost lowers 0.8-1 members/strain, by
It is yellowish green chimera in the floral leaf of blade, tissue culture difficulty is larger, and by adjusting the ratio of auxin and the basic element of cell division, blade is special
Sign is obvious, and the regularity of reproduction speed and seedling is greatly improved, preferably keeps original maternal character.
Embodiment
With reference to specific embodiment, the present invention is described in detail.
Embodiment 1
A kind of floral leaf Wu's wind grass fast breeding method based on tissue cultures, comprises the following steps：
(1) preparation of culture medium, the component and content of each stage culture medium of tissue culture are included：
1.1st, bud inducement cultivation base：MS+BA 3mg/L+NAA 0.3mg/L；
1.2nd, adventitious bud proliferation culture medium：MS+6-BA 1mg/L+NAA 0.1mg/L；
1.3rd, strong seedling culture base：MS+6-BA 0.2mg/L+NAA 0.02mg/L；
1.4th, root media：MS+NAA 0.1mg/L；
The pH5.8 of above-mentioned various culture mediums, sucrose 30g/L, agar 6g/L are added, cultivation temperature is 25 DEG C, and illumination is
1200Lx；
(2) tissue cultures of floral leaf Wu wind grass：
2.1st, the acquisition of sterilizable material
Floral leaf Wu's wind grass takes the budlet of aerial part in the month in spring 3-4, and 1h is rinsed after superclean bench with running water
On, 30s is soaked with 75% ethanol, 1 ‰ mercuric chloride soak 15min, aseptic water washing 5-6 times, and aseptic filter paper blots surface moisture content,
Take budlet base portion to be cut into 1cm length, be inoculated on bud inducement cultivation base；
2.2nd, the differentiation of bud and propagation
Budlet is inoculated on inducing culture after 30 days, and budlet base portion starts to expand, and yellow green projection is occurred, is further cultured for 25
Its visible obvious callus, continue culture 30 days, cut to be put on adventitious bud proliferation culture medium with bud callus and cultivate；
2.3rd, adventitious bud strong seedling culture
On adventitious bud proliferation culture medium, the only 3-4 strains of Multiple Buds every clump induced can extend, and remaining, which is in, downgrades
State, after being divided into little Cong or simple bud, strong seedling culture base is put into, adventitious bud is extended rapidly, and 2-3cm can be grown to after 20 days；
2.4th, culture of rootage
1.5-2cm plantlet is taken, root induction in root media of transferring, seedling base portion differentiates many after 20 days
The root restriction of white, can grow to 1-2cm after 30 days；
2.5th, hardening and transplanting
After culture of rootage 30 days, when root system length is to 1.5cm, the aseptic seedling of well developed root system robust growth, indoor corkage are selected
Hardening 3 days, root agar is cleaned after taking-up, hardening is tamed in greenhouse after 40 days, be i.e. transplanting is outdoor, gives rich water quality management, moves
It is 95% to plant survival rate.
Embodiment 2
A kind of floral leaf Wu's wind grass fast breeding method based on tissue cultures, comprises the following steps：
(1) preparation of culture medium, the component and content of each stage culture medium of tissue culture are included：
1.1st, bud inducement cultivation base：MS+BA 1mg/L+NAA 0.1mg/L；
1.2nd, adventitious bud proliferation culture medium：MS+6-BA 0.5mg/L+NAA 0.05mg/L；
1.3rd, strong seedling culture base：MS+6-BA 0.1mg/L+NAA 0.01mg/L；
1.4th, root media：MS+NAA 0.05mg/L；
The pH5.8 of above-mentioned various culture mediums, sucrose 30g/L, agar 6g/L are added, cultivation temperature is 24 DEG C, and illumination is
1100Lx；
(2) tissue cultures of floral leaf Wu wind grass：
2.1st, the acquisition of sterilizable material
Floral leaf Wu's wind grass takes the budlet of aerial part in the month in spring 3-4, and 1h is rinsed after superclean bench with running water
On, 30s is soaked with 75% ethanol, 1 ‰ mercuric chloride soak 15min, aseptic water washing 5-6 times, and aseptic filter paper blots surface moisture content,
Take budlet base portion to be cut into 1cm length, be inoculated on bud inducement cultivation base；
2.2nd, the differentiation of bud and propagation
Budlet is inoculated on inducing culture after 30 days, and budlet base portion starts to expand, and yellow green projection is occurred, is further cultured for 25
Its visible obvious callus, continue culture 30 days, cut to be put on adventitious bud proliferation culture medium with bud callus and cultivate；
2.3rd, adventitious bud strong seedling culture
On adventitious bud proliferation culture medium, the only 3-4 strains of Multiple Buds every clump induced can extend, and remaining, which is in, downgrades
State, after being divided into little Cong or simple bud, strong seedling culture base is put into, adventitious bud is extended rapidly, and 2-3cm can be grown to after 20 days；
2.4th, culture of rootage
1.5-2cm plantlet is taken, root induction in root media of transferring, seedling base portion differentiates many after 20 days
The root restriction of white, can grow to 1-2cm after 30 days；
2.5th, hardening and transplanting
After culture of rootage 28 days, when root system length is to 1cm, the aseptic seedling of well developed root system robust growth, indoor corkage refining are selected
Seedling 3 days, root agar is cleaned after taking-up, hardening is tamed in greenhouse after 40 days, be i.e. transplanting is outdoor, gives rich water quality management, transplants
Survival rate is 92%.
Embodiment 3
A kind of floral leaf Wu's wind grass fast breeding method based on tissue cultures, comprises the following steps：
(1) preparation of culture medium, the component and content of each stage culture medium of tissue culture are included：
1.1st, bud inducement cultivation base：MS+BA 3mg/L+NAA 0.3mg/L；
1.2nd, adventitious bud proliferation culture medium：MS+6-BA 1.5mg/L+NAA 0.15mg/L；
1.3rd, strong seedling culture base：MS+6-BA 0.3mg/L+NAA 0.03mg/L；
1.4th, root media：MS+NAA 0.15mg/L；
The pH5.8 of above-mentioned various culture mediums, sucrose 30g/L, agar 6g/L are added, cultivation temperature is 26 DEG C, and illumination is
1300Lx；
(2) tissue cultures of floral leaf Wu wind grass：
2.1st, the acquisition of sterilizable material
Floral leaf Wu's wind grass takes the budlet of aerial part in the month in spring 3-4, and 1h is rinsed after superclean bench with running water
On, 30s is soaked with 75% ethanol, 1 ‰ mercuric chloride soak 15min, aseptic water washing 5-6 times, and aseptic filter paper blots surface moisture content,
Take budlet base portion to be cut into 1cm length, be inoculated on bud inducement cultivation base；
2.2nd, the differentiation of bud and propagation
Budlet is inoculated on inducing culture after 30 days, and budlet base portion starts to expand, and yellow green projection is occurred, is further cultured for 25
Its visible obvious callus, continue culture 30 days, cut to be put on adventitious bud proliferation culture medium with bud callus and cultivate；
2.3rd, adventitious bud strong seedling culture
On adventitious bud proliferation culture medium, the only 3-4 strains of Multiple Buds every clump induced can extend, and remaining, which is in, downgrades
State, after being divided into little Cong or simple bud, strong seedling culture base is put into, adventitious bud is extended rapidly, and 2-3cm can be grown to after 20 days；
2.4th, culture of rootage
1.5-2cm plantlet is taken, root induction in root media of transferring, seedling base portion differentiates many after 20 days
The root restriction of white, can grow to 1-2cm after 30 days；
2.5th, hardening and transplanting
After culture of rootage 32 days, when root system length is to 2cm, the aseptic seedling of well developed root system robust growth, indoor corkage refining are selected
Seedling 3 days, root agar is cleaned after taking-up, hardening is tamed in greenhouse after 40 days, be i.e. transplanting is outdoor, gives rich water quality management, transplants
Survival rate is 93%.
The above-mentioned description to embodiment is understood that for ease of those skilled in the art and using invention.
Person skilled in the art obviously can easily make various modifications to these embodiments, and described herein general
Principle is applied in other embodiment without by performing creative labour.Therefore, the invention is not restricted to above-described embodiment, ability
Field technique personnel do not depart from improvement that scope made and modification all should be the present invention's according to the announcement of the present invention
Within protection domain.
Claims (5)
1. a kind of floral leaf Wu's wind grass fast breeding method based on tissue cultures, it is characterised in that comprise the following steps：
(1) preparation of culture medium, the component and content of each stage culture medium of tissue culture are included：
1.1st, bud inducement cultivation base：MS+BA 1-3mg/L+NAA 0.1-0.3mg/L；
1.2nd, adventitious bud proliferation culture medium：MS+6-BA 0.5-1.5mg/L+NAA 0.05-0.15mg/L；
1.3rd, strong seedling culture base：MS+6-BA 0.1-0.3mg/L+NAA 0.01-0.03mg/L；
1.4th, root media：MS+NAA 0.05-0.15mg/L；
The pH5.8 of above-mentioned various culture mediums, sucrose 30g/L, agar 6g/L are added, cultivation temperature is 25 ± 1 DEG C, and illumination is
1100-1300Lx；
(2) tissue cultures of floral leaf Wu wind grass：
2.1st, the acquisition of sterilizable material
Floral leaf Wu's wind grass takes the budlet of aerial part in the month in spring 3-4, rinses 1h after on superclean bench with running water, uses
75% ethanol immersion 30s, 1 ‰ mercuric chloride immersion 15min, aseptic water washing 5-6 times, aseptic filter paper blot surface moisture content, taken small
1cm length is cut into bastem portion, is inoculated on bud inducement cultivation base；
2.2nd, the differentiation of bud and propagation
Budlet is inoculated on inducing culture after 30 days, and budlet base portion starts to expand, and yellow green projection occurs, and being further cultured for 25 days can
See obvious callus, continue culture 30 days, cut to be put on adventitious bud proliferation culture medium with bud callus and cultivate；
2.3rd, adventitious bud strong seedling culture
On adventitious bud proliferation culture medium, the only 3-4 strains of Multiple Buds every clump induced can extend, and remaining, which is in, downgrades shape
State, after being divided into little Cong or simple bud, strong seedling culture base is put into, adventitious bud is extended rapidly, and 2-3cm can be grown to after 20 days；
2.4th, culture of rootage
1.5-2cm plantlet is taken, root induction in root media of transferring, seedling base portion differentiates many whites after 20 days
Root restriction, can be grown to 1-2cm after 30 days；
2.5th, hardening and transplanting
After culture of rootage 28-32 days, when root system length is to 1-2cm, the aseptic seedling of well developed root system robust growth, indoor corkage refining are selected
Seedling 3 days, root agar is cleaned after taking-up, hardening is tamed in greenhouse after 40 days, be i.e. transplanting is outdoor, gives rich water quality management, transplants
Survival rate is more than 90%.
A kind of 2. floral leaf Wu's wind grass fast breeding method based on tissue cultures according to claim 1, it is characterised in that
The component and content of described bud inducement cultivation base be：MS+BA 2mg/L+NAA 0.2mg/L.
A kind of 3. floral leaf Wu's wind grass fast breeding method based on tissue cultures according to claim 1, it is characterised in that
The component and content of described adventitious bud proliferation culture medium be：MS+6-BA 1mg/L+NAA 0.1mg/L.
A kind of 4. floral leaf Wu's wind grass fast breeding method based on tissue cultures according to claim 1, it is characterised in that
The component and content of described strong seedling culture base be：MS+6-BA 0.2mg/L+NAA 0.02mg/L.
A kind of 5. floral leaf Wu's wind grass fast breeding method based on tissue cultures according to claim 1, it is characterised in that
The component and content of described root media be：MS+NAA 0.1mg/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110537490A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园林植物研究所有限公司 | method for rapidly breeding floral leaf illicium verum discs through tissue culture |
| CN110537491A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园艺有限公司 | method for rapid breeding of African dual-color wild iris through tissue culture |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102273404A (en) * | 2010-06-09 | 2011-12-14 | 上海上房园艺有限公司 | Plant culturing method of spot Farfugium |
-
2016
- 2016-09-27 CN CN201610854285.4A patent/CN107864854A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102273404A (en) * | 2010-06-09 | 2011-12-14 | 上海上房园艺有限公司 | Plant culturing method of spot Farfugium |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110537490A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园林植物研究所有限公司 | method for rapidly breeding floral leaf illicium verum discs through tissue culture |
| CN110537491A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园艺有限公司 | method for rapid breeding of African dual-color wild iris through tissue culture |
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