CN107926710A - A kind of rapid propagation method of wild paint tissue culture - Google Patents
A kind of rapid propagation method of wild paint tissue culture Download PDFInfo
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- CN107926710A CN107926710A CN201711405602.5A CN201711405602A CN107926710A CN 107926710 A CN107926710 A CN 107926710A CN 201711405602 A CN201711405602 A CN 201711405602A CN 107926710 A CN107926710 A CN 107926710A
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- wild
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- seedling
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Classifications
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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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The invention belongs to technical field of plant propagation, discloses a kind of rapid propagation method of wild paint tissue culture, NAA, 6 BA, GA3 and AC are added into WPM culture mediums；Resting bud after disinfection is seeded in culture medium, and it is stem with bud to cultivate to Dormant Buds；It is seeded in the WPM culture mediums containing NAA, 6 BA and AC, Multiplying culture, obtains wild paint seedling；With the WPM culture mediums containing NAA, 6 BA, GA3 and AC, wild paint seedling is cultivated；The good wild paint seedling of growth conditions is chosen, using the White culture mediums containing IBA, 6 BA and AC, culture is to taking root；The addition of IBA is 2.0~4.0mg/L, the addition of 6 BA is 0~1.5mg/L, the addition of AC is 0.5~1.5g/L.The wild paint that the present invention is bred grows new root, survival rate more than 80% in 15 20 days.
Description
Technical field
The invention belongs to technical field of plant propagation, more particularly to a kind of rapid propagation method of wild paint tissue culture.
Background technology
Deciduous tree or dungarunga, up to 10 meters；Sprig is sturdy.Imparipinnate leaf alternate, Chang Jisheng sprigs top are small
Leaf is to raw or closely to life, hard papery to thin keratin, and long round shape is oval, wealthy lanceolar or ovum shape lanceolar.The long 7- of panicle
15 centimetres, flower yellow green.It is born in the woods of height above sea level (150-) 300-1500 (- 2500) rice.Root, leaf and fruit are used as medicine, and have heat-clearing solution
Poison, dissipate the effect such as stasis of blood myogenic；Trees with Important Economic value.Chief value：Medical value：Root, leaf and fruit are used as medicine, and are had clear
Thermal detoxification, dissipate stasis of blood myogenic, hemostasis, the effect of desinsection, controls injury and bone fracture, eczema sore, venomous snake bite, and can control hematuria, metrorrhagia, white
The diseases such as band, traumatic hemorrhage, metroptosis.Economic value：Seed oil can soapmaking or blend drying oil paint.The lacquer tree fat of pericarp can
Candle processed, plaster and pomade etc..Bark can carry tannin extract.Trunk lotion can be used for raw lacquer.Timber hard and compact, can be used as fine workmanship
Material.In recent years, Mianyang City Pingwu County, Beichuanqiangzu Autonomous County development universe tourism, make colored Pingwu, colored Beichuan.Open country paint
It is Mianyang City indigenous tree, has higher economic value, and autumn red autumnal leaves effect is preferable.But the differentiation of seminal propagation seedling is serious；
Between Wild plant or the plant of seminal propagation, red autumnal leaves time, degree difference are larger；Since seed is by male parent and maternal biography
Powder fertilization, genetic recombination are formed.So with parent, there are larger hereditary difference.Progeny character is widely different.After growing up, have
Leaf is not red, or the red time is than later.It can not carry out popularizing planting.Need by vegetative propagation by the red autumnal leaves time it is relatively early,
More attractive maternal plant quickly breeds, and the short time produces a large amount of seedlings, and retains gene and be basically unchanged.
In conclusion problem existing in the prior art is：There are seedling differentiation is serious for open country paint seminal propagation；Wild plant or
Between the plant of seminal propagation, red autumnal leaves time, degree difference are larger.
The content of the invention
In view of the problems of the existing technology, the present invention provides a kind of rapid propagation method of wild paint tissue culture.
The present invention is achieved in that a kind of rapid propagation method of wild paint tissue culture, the quick breeding of the wild paint tissue culture
Method comprises the following steps：
Step 1, to WPM culture mediums (xylophyta with add in culture medium (WPM, Woody Plant medium) NAA,
6-BA, GA3 and AC, it is 5.8~6.0 to adjust Medium's PH Value, and sterilize 20~25min in 120~125 DEG C, is cooled to room temperature；Disappear
Resting bud after poison is seeded in culture medium, and under the conditions of 24 ± 1 DEG C, 2000~2100lx, 8~10h/d of illumination, is cultivated to not
It is stem with bud that dormancy bud, which is sprouted,；NAA additions are 0~0.4mg/L, 6-BA additions are 2.0~4.0mg/L, GA3 additions are
0~2.0mg/L, AC additions are 1.0~1.5g/L；
Step 2, is seeded in the WPM culture mediums containing NAA, 6-BA and AC, 24 ± 1 DEG C, 2000~2100lx conditions
Under, 8~10h/d of illumination, Multiplying culture, obtains wild paint seedling；NAA additions add for 0.2~0.6mg/L, 6-BA in WPM culture mediums
Enter amount be 1.0~4.0mg/L, AC additions be 0.5~1.5g/L；
Step 3, with the WPM culture mediums containing NAA, 6-BA, GA3 and AC, in 24 ± 1 DEG C, 2000~2100lx conditions
Under, 8~10h/d of illumination, cultivates wild paint seedling；NAA additions are 0.5~1.5mg/L, 6-BA additions be 0.2~1.0mg/L,
GA3 additions are 0.5~1.5mg/L, AC additions are 0.5~1.5g/L；
Step 4, chooses the good wild paint seedling of growth conditions, using the White culture mediums containing IBA, 6-BA and AC, in
24 ± 1 DEG C, under the conditions of 2000~2100lx, 8~10h/d of illumination, culture is to taking root；The addition of IBA for 2.0~4.0mg/L,
The addition of 6-BA is 0~1.5mg/L, the addition of AC is 0.5~1.5g/L.
Further, need to carry out before the step 1：
(1) after fall foliage before, sprouting, it is explant material to gather the resting bud that 1~2 year raw wild paint branch is not sprouted；
(2) resting bud is placed in the washing powder solution that concentration is 3~5g/mL and soaks 2~3min, repair resting bud, only
The bud-end of surplus 0.3~0.5cm, soaks 10~30s with 75% ethanol, then soaks 1~7min with 0.01% mercuric chloride solution；Or only make
With 0.01% mercuric chloride solution immersion resting bud, 1~8min.
Further, need to carry out after the step 4：Tissue-cultured seedling Jing Guo hardening is taken out from bottle, is cleaned with clear water
Culture medium on root system, soaks 10min with the solution for the Bravo 0.1% that mass fraction is 80%；Tissue-cultured seedling is transplanted to matrix
In, according to weight ratio turf：Perlite=1：1；Cultivation matrix will pour permeable before transplanting, and with new 0.025% liquid of phytomycin
Uniformly sprinkling stromal surface.
Further, water spray at initial stage moisturizing, relative humidity are maintained at more than 80%, and transplanting gradually reduces relative humidity after 1 week；
18-25 DEG C of temperature and 50% intensity of illumination.
Another object of the present invention is to provide a kind of wild paint bred using the wild rapid propagation method for painting tissue culture.
Advantages of the present invention and good effect are：1. ensure seedling fine quality.Since seed is by male parent and female parent
Pollination fertilization, genetic recombination are formed.So there are larger hereditary difference by filial generation and parent.It cannot be guaranteed that obtain excellent heredity
Gene.And the present invention solves the problems, such as this.2. ensure the uniformity of seedling.Seminal propagation progeny character is widely different.After growing up,
Some leaves are not red, or the red time is than later.It can not carry out popularizing planting.The seedling that the present invention cultivates, as long as tissue culture material
Expect that source is consistent.It can ensure hereditary consistent.Under identical planting conditions, performance is consistent.3. the present invention can be short by tissue-culturing rapid propagation
A large amount of seedling are obtained in time.First batch of material 60d or so since second batch, can save materials, initial incubation with seedling,
Directly cultivated since Proliferation, Differentiation, 50d can be with seedling.In theory, can be with the short time as long as instrument and equipment and human hand are enough
Obtain an infinite number of nursery stock.Influenced from raw material, weather etc..The elite plant that the present invention will can select, passes through group
Training obtains a large amount of seedling in fast numerous short time, and can guarantee that genetic consistent.
Brief description of the drawings
Fig. 1 is the rapid propagation method flow chart of wild paint tissue culture provided in an embodiment of the present invention.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Deciduous tree or dungarunga root, leaf and fruit are used as medicine, and are had clearing heat and detoxicating, scattered stasis of blood myogenic, hemostasis, the effect of desinsection, are controlled and fall
Fracture, eczema sore, venomous snake bite are beaten, and the diseases such as hematuria, metrorrhagia, leukorrhea, traumatic hemorrhage, metroptosis can be controlled.Seed oil can
Soapmaking or blending drying oil paint；The lacquer tree fat of pericarp can candle processed, plaster and pomade etc.；Bark can carry tannin extract；Trunk lotion
It can be used for raw lacquer；Timber hard and compact, can make small timber.
The application principle of the present invention is explained in detail below in conjunction with the accompanying drawings.
As shown in Figure 1, the rapid propagation method of wild paint tissue culture provided in an embodiment of the present invention comprises the following steps：
S101：After fall foliage, sprout before, it is explant material to gather the resting bud that 1~2 year raw wild paint branch is not sprouted
Material；
S102：Resting bud is placed in the washing powder solution that concentration is 3~5g/mL and soaks 2~3min, repairs resting bud,
The bud-end of only surplus 0.3~0.5cm, soaks 10~30s with 75% ethanol, then soaks 1~7min with 0.01% mercuric chloride solution；Or only
Use 0.01% mercuric chloride solution immersion resting bud, 1~8min；
S103：NAA, 6-BA, GA3 and AC are added into WPM culture mediums, it is 5.8~6.0 to adjust Medium's PH Value, in 120
~125 DEG C of 20~25min of sterilizing, are cooled to room temperature, spare；Then the resting bud after disinfection is seeded in culture medium, in 24
± 1 DEG C, under the conditions of 2000~2100lx, 8~10h/d of illumination, it is stem with bud to cultivate to Dormant Buds；Wherein, NAA is added
Measure as 0~0.4mg/L, 6-BA additions are 2.0~4.0mg/L, GA3 additions are 0~2.0mg/L, AC additions be 1.0~
1.5g/L；
S104：It is seeded in the WPM culture mediums containing NAA, 6-BA and AC, in 24 ± 1 DEG C, 2000~2100lx conditions
Under, 8~10h/d of illumination, Multiplying culture, obtains wild paint seedling；NAA additions add for 0.2~0.6mg/L, 6-BA in WPM culture mediums
Enter amount be 1.0~4.0mg/L, AC additions be 0.5~1.5g/L；
S105：With the WPM culture mediums containing NAA, 6-BA, GA3 and AC, under the conditions of 24 ± 1 DEG C, 2000~2100lx,
8~10h/d of illumination, cultivates wild paint seedling；Wherein, NAA additions be 0.5~1.5mg/L, 6-BA additions be 0.2~1.0mg/
L, GA3 additions be 0.5~1.5mg/L, AC additions be 0.5~1.5g/L；
S106：The good wild paint seedling of growth conditions is chosen, using the White culture mediums containing IBA, 6-BA and AC, in 24
± 1 DEG C, under the conditions of 2000~2100lx, 8~10h/d of illumination, culture is to taking root；Wherein, the addition of IBA for 2.0~
The addition of 4.0mg/L, 6-BA are 0~1.5mg/L, the addition of AC is 0.5~1.5g/L；
S107：Tissue-cultured seedling Jing Guo abundant hardening is carefully taken out from bottle, the culture medium on root system is cleaned with clear water, is used
Mass fraction is that the solution of 80% Bravo 0.1% soaks 10min, and to improve nursery stock disease resistance, tissue-cultured seedling is carefully transplanted
(the turf into matrix：Perlite=1：1)；Cultivation matrix will pour permeable before transplanting, and uniform with new 0.025% liquid of phytomycin
Spray stromal surface；The generation of the damping-off of seedling can effectively be prevented；Pay attention to initial stage spray water moisturizing, relative humidity be maintained at 80% with
On, 1 Zhou Houke of transplanting gradually reduces relative humidity；Temperature is maintained at 18-25 DEG C and suitable intensity of illumination, is tissue-cultured seedling transplanting
Suitable environmental condition.
The application principle of the present invention is further described with reference to experiment.
The hardening time：6d, 8d, 10d, 12d, 14d5 processing of bottle refining time are respectively adopted in experiment.Timing statistics are transplanting
After one month；Bottle refining time 10d is optimal.
Seen by the survival rate after the tissue culture transplantation of seedlings to the different hardening times and the investigation result of seedling-height growth situation,
The different hardening times are to the survival rate of nursery stock and the influence significant difference of Seedling height.Tissue-cultured seedling bottle refining chronergy it is relatively good be
10d, survival rate more than 80%, Seedling height has also reached more than 5cm, and the wild paint tissue-cultured seedling of this explanation has had after 10d hardenings
There is stronger adaptation external environment ability.The tissue-cultured seedling of hardening 6d, leaf color is light green, and easily dehydration is wilted, and shows obvious
Hardening it is insufficient.And the nursery stock that the hardening time is 14d, root system is long, and nest root is be easy to cause during transplanting, influences transplanting effect
Fruit.In addition hardening overlong time, root system color browning, shows a degree of aging, is unfavorable for the formation of new root.
Hardening matrix presses 1 according to weight ratio turf and perlite：1 ratio is prepared.Generally 1 grows new root in 5-20 days.Into
Motility rate is up to 8 more than 0%.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should all be included in the protection scope of the present invention.
Claims (5)
- A kind of 1. rapid propagation method of wild paint tissue culture, it is characterised in that the rapid propagation method of the wild paint tissue culture include with Lower step：Step 1, NAA, 6-BA, GA3 and AC are added into WPM culture mediums, and it is 5.8~6.0 to adjust Medium's PH Value, in 120~ 125 DEG C of 20~25min of sterilizing, are cooled to room temperature；Resting bud after disinfection is seeded in culture medium, in 24 ± 1 DEG C, 2000~ Under the conditions of 2100lx, 8~10h/d of illumination, it is stem with bud to cultivate to Dormant Buds；NAA additions are 0~0.4mg/L, 6- BA additions are 2.0~4.0mg/L, GA3 additions are 0~2.0mg/L, AC additions are 1.0~1.5g/L；Step 2, is seeded in the WPM culture mediums containing NAA, 6-BA and AC, 24 ± 1 DEG C, under the conditions of 2000~2100lx, light According to 8~10h/d, Multiplying culture, obtains wild paint seedling；NAA additions are 0.2~0.6mg/L, 6-BA additions in WPM culture mediums It is 0.5~1.5g/L for 1.0~4.0mg/L, AC additions；Step 3, with the WPM culture mediums containing NAA, 6-BA, GA3 and AC, under the conditions of 24 ± 1 DEG C, 2000~2100lx, light According to 8~10h/d, wild paint seedling is cultivated；NAA additions are 0.5~1.5mg/L, 6-BA additions are 0.2~1.0mg/L, GA3 adds Enter amount be 0.5~1.5mg/L, AC additions be 0.5~1.5g/L；Step 4, chooses the good wild paint seedling of growth conditions, using the White culture mediums containing IBA, 6-BA and AC, in 24 ± 1 DEG C, under the conditions of 2000~2100lx, 8~10h/d of illumination, culture is to taking root；The addition of IBA is 2.0~4.0mg/L, 6-BA Addition be 0~1.5mg/L, the addition of AC be 0.5~1.5g/L.
- 2. the rapid propagation method of paint tissue culture as claimed in claim 1 wild, it is characterised in that needed before the step 1 into OK：(1) after fall foliage before, sprouting, it is explant material to gather the resting bud that 1~2 year raw wild paint branch is not sprouted；(2) resting bud is placed in the washing powder solution that concentration is 3~5g/mL and soaks 2~3min, repair resting bud, only surplus 0.3 The bud-end of~0.5cm, soaks 10~30s with 75% ethanol, then soaks 1~7min with 0.01% mercuric chloride solution；Or it is used only 0.01% mercuric chloride solution immersion resting bud, 1~8min.
- 3. the rapid propagation method of paint tissue culture as claimed in claim 1 wild, it is characterised in that needed after the step 4 into OK：Tissue-cultured seedling Jing Guo hardening is taken out from bottle, the culture medium on root system is cleaned with clear water, is 8 0% with mass fraction The solution of Bravo 0.1% soaks 1 0min；Tissue-cultured seedling is transplanted in matrix, according to weight ratio turf：Perlite=1：1；Move Cultivation matrix will pour permeable before cultivation, and uniformly spray stromal surface with new 0.025% liquid of phytomycin.
- 4. the rapid propagation method of wild paint tissue culture as claimed in claim 3, it is characterised in that moisturizing of spraying water initial stage, it is relatively wet Degree is maintained at 8 more than 0%, and transplanting gradually reduces relative humidity after 1 week；18-25 DEG C of temperature and 50% intensity of illumination.
- 5. a kind of usage right requires the wild paint of the rapid propagation method breeding of wild paint tissue culture described in 1~4 any one.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711405602.5A CN107926710A (en) | 2017-12-22 | 2017-12-22 | A kind of rapid propagation method of wild paint tissue culture |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711405602.5A CN107926710A (en) | 2017-12-22 | 2017-12-22 | A kind of rapid propagation method of wild paint tissue culture |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104012417A (en) * | 2014-07-02 | 2014-09-03 | 西南林业大学 | High-efficiency and rapid micropropagation method for toxicodendron vernicifluum |
| CN105660398A (en) * | 2016-01-14 | 2016-06-15 | 湖北省林业科学研究院 | Tissue culture rapid-propagation method of Maoba wild sumac |
| CN106818476A (en) * | 2017-01-19 | 2017-06-13 | 西南林业大学 | The efficient quick propagation method of natural Triploid Lacquer Tree |
| CN106818477A (en) * | 2017-01-19 | 2017-06-13 | 西南林业大学 | The efficient quick propagation method of japanese lacquer tree |
-
2017
- 2017-12-22 CN CN201711405602.5A patent/CN107926710A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104012417A (en) * | 2014-07-02 | 2014-09-03 | 西南林业大学 | High-efficiency and rapid micropropagation method for toxicodendron vernicifluum |
| CN105660398A (en) * | 2016-01-14 | 2016-06-15 | 湖北省林业科学研究院 | Tissue culture rapid-propagation method of Maoba wild sumac |
| CN106818476A (en) * | 2017-01-19 | 2017-06-13 | 西南林业大学 | The efficient quick propagation method of natural Triploid Lacquer Tree |
| CN106818477A (en) * | 2017-01-19 | 2017-06-13 | 西南林业大学 | The efficient quick propagation method of japanese lacquer tree |
Non-Patent Citations (2)
| Title |
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| 引田裕之等: ""漆树萌芽组织培养试管苗的培育"", 《广西林业科技》 * |
| 罗一然等: ""小木漆组织培养技术研究"", 《西南林业大学学报》 * |
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