CN106718912B - A kind of tissue culture and rapid propagation method of lycium ruthenicum - Google Patents
A kind of tissue culture and rapid propagation method of lycium ruthenicum Download PDFInfo
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- CN106718912B CN106718912B CN201611205895.8A CN201611205895A CN106718912B CN 106718912 B CN106718912 B CN 106718912B CN 201611205895 A CN201611205895 A CN 201611205895A CN 106718912 B CN106718912 B CN 106718912B
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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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- 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 present invention provides a kind of tissue culture and rapid propagation methods of lycium ruthenicum, belong to breeding plant cultivation field.This method comprises the step of: (1) obtaining aseptic seedling；(2) Fiber differentiation；(3) Multiplying culture；(4) culture of rootage；(5) nutrition is cultivated and looked after.Using lycium ruthenicum shoot survival percent prepared by the present invention, the breeding cycle is short, and seedling quality is good for 95%, and emergence rate is high；Without cultivating stock and scion, occupied area is greatly reduced, manpower and management cost are reduced.
Description
Technical field
The invention belongs to breeding plant cultivation fields, and in particular to a kind of tissue culture and rapid propagation method of lycium ruthenicum.
Background technique
Black fruit fructus lycii, the more quil shrubs of Solanaceae, Lycium are distributed in high mountain sarin, salinization sand ground, river and lake bank, dry river
It is the western distinctive desert medicinal plant kind in China in bed, Desert Riparian Forest.Wild black fruit fructus lycii adaptability is very strong, energy
Restrain oneself 38.5 celsius temperatures, cold resistance is also very strong, and it is drought-resistant without freeze injury under -25.6 degrees Celsius, it in desert remains to give birth to
It is long.It is intolerant tree species, develops healthy and strong under full exposure, lower growth is thin and delicate, and flowers and fruits are few giving shade, and requires not tight, salt tolerant to soil
Alkali, it is drought-resistant.
Although lycium ruthenicum can mostly use greatly the routine side of seminal propagation and cutting propagation with artificial growth at present
Formula plantation, is not able to satisfy the market demand still, and tissue cultures are a kind of efficient vegetative manners, reproductive efficiency is high and cost compared with
It is low, it is to solve the fast numerous effective means of lycium ruthenicum choiceness.
Summary of the invention
The technical problem to be solved by the present invention is in view of the shortcomings of the prior art or defect, provide a kind of lycium ruthenicum
Tissue culture and rapid propagation method.
To achieve the goals above, the present invention is achieved by the following scheme:
A kind of tissue culture and rapid propagation method of lycium ruthenicum, comprising the following steps:
(1) aseptic seedling is obtained
Lycium ruthenicum seed is taken, 6~10h is impregnated in 30 DEG C of water, in 75% ethanol postincubation on superclean bench after taking-up
30s impregnates 3~8min with the liquor natrii hypochloritis that mass concentration is 10%, and with aseptic water washing 3~4 times, filter paper blots surface
Moisture accesses seed in 15~25g/L+ of DKW+ sucrose agar 4~5g/L culture medium, in pH5.8, illumination 2000lx, temperature
It is cultivated under conditions of 25 DEG C, grows sterile bud；
(2) Fiber differentiation
Sterile sprout is accessed into DKW+0.5~1mgL-1KT+0.1~0.3mgL-1IBA+10~15gL-1Sucrose+3
~5gL-1In agar medium, Fiber differentiation under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature grows and grows thickly
Bud；
(3) Multiplying culture
Multiple Buds are cut into single plant bud, access DKW+0.7~1.2mgL-16-BA+0.2~0.3mgL-1IBA+10~
15g·L-1Sucrose+3~5gL of agar-1In culture medium, training is proliferated under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature
It supports；
(4) culture of rootage
When single plant bud grows to 2~3cm, it is forwarded to: DKW+0.2~0.3mgL-1IBA+0.02~0.03mgL- 1NAA+10~15gL-1Sucrose+4~5gL of agar-1In culture medium, in pH5.8, illumination 2000lx, 25 DEG C of temperature of condition
Lower culture of rootage；
(5) nutrition is cultivated and looked after
After culture of rootage January, it is placed under culturing room's natural light and opens culture bottle bottleneck white silk seedling 5~7 days, it is then careful to take out
Seedling cleans root culture medium, and carbendazim soaks root, moves into leaf mould: rural area soil: vermiculite: perlite=1:1:1:1 matrix
In, it is primary every spray in 10 days with 1000 times of potassium dihydrogen phosphate foliage-sprays after slow seedling, nursery, which is moved into, after culture 50~60 days carries out
Maintenance management.
Advantageous effects of the invention: using lycium ruthenicum shoot survival percent prepared by the present invention, the breeding cycle is short for 95%,
Seedling quality is good, and emergence rate is high；Without cultivating stock and scion, occupied area is greatly reduced, manpower and management cost are reduced.
Specific embodiment
It further elaborates, does not constitute to of the invention any to technical solution of the present invention below with reference to embodiment
Range limitation.
Embodiment 1
Lycium ruthenicum seed is taken, impregnates 6h in 30 DEG C of water, in 75% ethanol postincubation 30s on superclean bench after taking-up, is used
The liquor natrii hypochloritis that mass concentration is 10% impregnates 3min, and with aseptic water washing 3 times, filter paper blots surface moisture, by seed
It accesses in DKW+ sucrose 15g/L+ agar 4g/L culture medium, is cultivated under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature, it is raw
Grow sterile bud.Sterile sprout is accessed into DKW+0.5mgL-1KT+0.1mg·L-1IBA+10g·L-1Sucrose+3gL-1Agar
In culture medium, Fiber differentiation under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature grows Multiple Buds.Multiple Buds are cut
At single plant bud, DKW+0.7mgL is accessed-16-BA+0.2mg·L-1IBA+10g·L-1Sucrose+agar 3gL-1In culture medium,
Multiplying culture under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature.When single plant bud grows to 2cm, it is forwarded to: DKW+
0.2mg·L-1IBA+0.02mg·L-1NAA+10~15gL-1Sucrose+agar 4gL-1In culture medium, in pH5.8, illumination
2000lx, culture of rootage under conditions of 25 DEG C of temperature.After culture of rootage January, it is placed in opening culture bottle bottle under culturing room's natural light
Mouth is practiced seedling 5 days, and seedling is then carefully taken out, and cleans root culture medium, and carbendazim leaching root moves into leaf mould: rural area soil: vermiculite:
It is primary every spray in 10 days with 1000 times of potassium dihydrogen phosphate foliage-sprays after slow seedling in perlite=1:1:1:1 matrix, culture
Nursery is moved into after 50 days carries out maintenance management, transplanting survival rate 98%.
Embodiment 2
Lycium ruthenicum seed is taken, impregnates 10h in 30 DEG C of water, in 75% ethanol postincubation 30s on superclean bench after taking-up, is used
The liquor natrii hypochloritis that mass concentration is 10% impregnates 8min, and with aseptic water washing 4 times, filter paper blots surface moisture, by seed
It accesses in DKW+ sucrose 25g/L+ agar 5g/L culture medium, is cultivated under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature, it is raw
Grow sterile bud.Sterile sprout is accessed into DKW+1mgL-1KT+0.3mg·L-1IBA+15g·L-1Sucrose+5gL-1Agar training
It supports in base, Fiber differentiation under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature grows Multiple Buds.Multiple Buds are cut into
Single plant bud accesses DKW+1.2mgL-16-BA+0.3mg·L-1IBA+15g·L-1Sucrose+agar 5gL-1In culture medium,
PH5.8, illumination 2000lx, Multiplying culture under conditions of 25 DEG C of temperature.When single plant bud grows to 3cm, it is forwarded to: DKW+
0.3mg·L-1IBA+0.03mg·L-1NAA+15g·L-1Sucrose+agar 5gL-1In culture medium, in pH5.8, illumination
2000lx, culture of rootage under conditions of 25 DEG C of temperature.After culture of rootage January, it is placed in opening culture bottle bottle under culturing room's natural light
Mouth is practiced seedling 7 days, and seedling is then carefully taken out, and cleans root culture medium, and carbendazim leaching root moves into leaf mould: rural area soil: vermiculite:
It is primary every spray in 10 days with 1000 times of potassium dihydrogen phosphate foliage-sprays after slow seedling in perlite=1:1:1:1 matrix, culture
Nursery is moved into after 60 days carries out maintenance management, transplanting survival rate 95%.
Embodiment 3
Lycium ruthenicum seed is taken, impregnates 8h in 30 DEG C of water, in 75% ethanol postincubation 30s on superclean bench after taking-up, is used
The liquor natrii hypochloritis that mass concentration is 10% impregnates 5min, and with aseptic water washing 4 times, filter paper blots surface moisture, by seed
It accesses in DKW+ sucrose 20g/L+ agar 4.5g/L culture medium, is cultivated under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature,
Grow sterile bud.Sterile sprout is accessed into DKW+75mgL-1KT+0.2mg·L-1IBA+12.5g·L-1Sucrose+4gL-1
In agar medium, Fiber differentiation under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature grows Multiple Buds.It will grow thickly
Bud is cut into single plant bud, accesses DKW+1mgL-16-BA+0.25mg·L-1IBA+12.5g·L-1Sucrose+agar 4gL-1Culture
In base, Multiplying culture under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature.When single plant bud grows to 3cm, it is forwarded to:
DKW+0.25mg·L-1IBA+0.025mg·L-1NAA+12.5g·L-1Sucrose+agar 4.5gL-1In culture medium, pH5.8,
Illumination 2000lx, culture of rootage under conditions of 25 DEG C of temperature.After culture of rootage January, it is placed under culturing room's natural light and opens culture
Then bottle bottleneck white silk seedling 5~7 days carefully takes out seedling, clean root culture medium, and carbendazim soaks root, moves into leaf mould: rural area
Soil: in perlite=1:1:1:1 matrix, with 1000 times of potassium dihydrogen phosphate foliage-sprays after slow seedling, one vermiculite: was sprayed every 10 days
It is secondary, nursery, which is moved into, after culture 50~60 days carries out maintenance management, transplanting survival rate 96%.
Claims (1)
1. a kind of tissue culture and rapid propagation method of lycium ruthenicum, which comprises the following steps:
(1) aseptic seedling is obtained
Lycium ruthenicum seed is taken, 6~10h is impregnated in 30 DEG C of water, in 75% ethanol postincubation 30s on superclean bench after taking-up, is used
The liquor natrii hypochloritis that mass concentration is 10% impregnates 3~8min, and with aseptic water washing 3~4 times, filter paper blots surface moisture,
Seed is accessed in 15~25g/L+ of DKW+ sucrose agar 4~5g/L culture medium, in pH5.8, illumination 2000lx, 25 DEG C of temperature
Under the conditions of cultivate, grow sterile bud；
(2) Fiber differentiation
Sterile sprout is accessed into DKW+0.5~1mgL-1KT+0.1~0.3mgL-1IBA+10~15gL-1Sucrose+3~
5g·L-1In agar medium, Fiber differentiation under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature grows Multiple Buds；
(3) Multiplying culture
Multiple Buds are cut into single plant bud, access DKW+0.7~1.2mgL-16-BA+0.2~0.3mgL-1IBA+10~15g
L-1Sucrose+3~5gL of agar-1In culture medium, Multiplying culture under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature；
(4) culture of rootage
When single plant bud grows to 2~3cm, it is forwarded to: DKW+0.2~0.3mgL-1IBA+0.02~0.03mgL-1NAA+
10~15gL-1Sucrose+4~5gL of agar-1It is raw under conditions of pH5.8, illumination 2000lx, 25 DEG C of temperature in culture medium
Root culture；
(5) nutrition is cultivated and looked after
After culture of rootage January, it is placed under culturing room's natural light and opens culture bottle bottleneck white silk seedling 5~7 days, it is then careful to take out children
Seedling cleans root culture medium, and carbendazim soaks root, moves into leaf mould: rural area soil: vermiculite: in perlite=1:1:1:1 matrix,
It is primary every spray in 10 days with 1000 times of potassium dihydrogen phosphate foliage-sprays after slow seedling, nursery, which is moved into, after culture 50~60 days is supported
Pillar reason.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104719165A (en) * | 2015-03-30 | 2015-06-24 | 四川禾木本业农林科技有限公司 | Rapid tissue culture method for lycium ruthenicum murr |
| CN105815221A (en) * | 2016-04-06 | 2016-08-03 | 沈阳农业大学 | Method for in-vitro rapid propagation of lycium ruthenicum by taking young seedlings as explant donors |
| CN105850733A (en) * | 2016-04-07 | 2016-08-17 | 甘肃省治沙研究所 | Lycium ruthenicum regenerated seedling cultivation method |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104719165A (en) * | 2015-03-30 | 2015-06-24 | 四川禾木本业农林科技有限公司 | Rapid tissue culture method for lycium ruthenicum murr |
| CN105815221A (en) * | 2016-04-06 | 2016-08-03 | 沈阳农业大学 | Method for in-vitro rapid propagation of lycium ruthenicum by taking young seedlings as explant donors |
| CN105850733A (en) * | 2016-04-07 | 2016-08-17 | 甘肃省治沙研究所 | Lycium ruthenicum regenerated seedling cultivation method |
Non-Patent Citations (5)
| Title |
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| 荒漠区药用植物黑果枸杞(Lycium ruthencium)的组织培养;王方琳等;《干旱区资源与环境》;20161015;第30卷(第10期);第104-109页 |
| 黑果枸杞的组织培养;浩仁塔本等;《植物生理学通讯》;20051031;第41卷(第5期);第631页 |
| 黑果枸杞的组织培养和植株再生;胡相伟等;《农业科技与信息》;20151231(第7期);第48-49页 |
| 黑果枸杞组织培养快繁技术研究;马彦军等;《林业科技通讯》;20151231(第6期);第26-28页 |
| 黑果枸杞组织培养技术;胡相伟等;《甘肃农业科技》;20151231(第5期);第73-74页 |
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Effective date of registration: 20181127 Address after: 318000 Feilong Village 532, Sanjia Street, Jiaojiang District, Taizhou City, Zhejiang Province Applicant after: Pan Zhiyuan Address before: 745000 225 North Street, Xifeng District, Qingyang, Gansu. Applicant before: Qingyang science and Technology Development Co., Ltd. |
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