CN108271691A - A method of carrying out tissue cultures using Kiwi berry stem - Google Patents
A method of carrying out tissue cultures using Kiwi berry stem Download PDFInfo
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- CN108271691A CN108271691A CN201810083297.0A CN201810083297A CN108271691A CN 108271691 A CN108271691 A CN 108271691A CN 201810083297 A CN201810083297 A CN 201810083297A CN 108271691 A CN108271691 A CN 108271691A
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- kiwi berry
- culture
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- root
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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 relates to a kind of methods carrying out tissue cultures using Kiwi berry stem.3cm or more is reached after proliferation when root induction, the plant with 3 or more cotyledons is inserted into the matrix inside special equipment of taking root, which is MS+ vermiculites, light application time 16h, 25 ± 2 DEG C of temperature, humidity 80%, gas concentration lwevel 900ppm 1000ppm, 1000 grades of culturing room's cleanliness factor or more.Culture forms the Kiwi berry intact plant of well developed root system after 20 days.The present invention uses vegetative manner, and maternal merit is made completely to retain.And the problem of woody tissue-cultured seedling quality of rooting difference is breached, tissue-cultured seedling well developed root system, completely, survival rate are high, without hardening.
Description
Technical field
It is the present invention relates to field of plant tissue culture technique, more particularly to a kind of to carry out tissue cultures using Kiwi berry stem
Method.
Background technology
Kiwi berry (scientific name：Actinidia chinensis Planch), it is perennial bejuco, Kiwi berry
Soft texture, sweet mouthfeel.Taste is described as the mixing of strawberry, banana, pineapple three.Kiwi berry remove containing actinidine,
The organic matters and calcium, potassium, selenium, zinc, germanium etc. such as proteolytic enzyme, tannin pectin and carbohydrate trace element and 17 kinds of needed by human body
Outside amino acid, also contain abundant vitamin C, grape acid, fructose, citric acid, malic acid, fat.
The propagation method of traditional Kiwi berry has sowing, cuttage, press strip and grafting etc..These method relative efficiencies are relatively low, mesh
Before, there are many methods using tissue cultures to carry out Kiwi berry breeding, such as：Patent CN102177847A discloses one kind
The factorial seedling-culturing method of tara vine, including the preparation of culture medium, the selection of explant and disinfection, inoculation, squamous subculture,
Sand bed transplants five steps.But this method needs to configure complicated component and culture medium configuration process requires stringent basic culture
Base, inducing culture and subculture medium, operating process is cumbersome, needs to put into more manpower and equipment cost, does not have wide
General applicability.But in the method for existing tissue cultures.There are many disadvantages, are all especially to need to contain in the stage of taking root
Sugar culture-medium, but sugar is pollution sources, the pollution rate in stage easy to increase of taking root, moreover, conventional method can only obtain when taking root
3-5 main root, and do not have root hair, as shown in Fig. 2, needing later stage hardening culture.
Invention content
For problems of the prior art, tissue training is carried out using Kiwi berry stem the object of the present invention is to provide a kind of
Foster method.
In order to solve the above-mentioned technical problem, the present invention is achieved by the following scheme：
Blade explant evoked callus method
1. acquiring the fresh cane of Kiwi berry, clear water wash clean.Cane is cut into the stem section of 4cm or so, at least retains 2 armpits
Bud, 0.1% mercuric chloride sterilizing 15m, 75% alcohol sterilizing 5m, sterile water wash 3-5 times.It is inserted into MS+ sucrose 30g/L+ agar 6g/L+
In the culture medium of 6-BA0.8mg/L+IBA0.5mg/L, illumination 2000-3000Lx, 14h, 25 ± 2 DEG C of temperature, humidity 70%, training
After supporting 25 days or so, adventitious bud is born at axillary bud position, obtains primary.
2. the adventitious bud formed on callus is inserted into MS+ sucrose 30g/L+ agar 6g/L by squamous subculture (Multiplying culture)
On the culture medium of+6-BA1.2mg/L+IAA0.8mg/L, illumination 2000-3000LX, 14h, 25 ± 2 DEG C of temperature, humidity 70%, training
It supports 25 days or so, 3-5 times of appreciation rate.
3. root induction after proliferation 3cm or more is reached, the plant with 3 or more cotyledons is inserted into special take root in equipment
In the matrix in face, which is MS+ vermiculites, light application time 16h, 25 ± 2 DEG C of temperature, humidity 80%, gas concentration lwevel
900ppm-1000ppm, 1000 grades of culturing room's cleanliness factor or more.Culture forms well developed root system Kiwi berry after 20 days is completely planted
Strain.
MS+ vermiculites are that every liter of MS solution adds 800g vermiculites.
4. a Kiwi berry intact plant is transplanted to inside degradation bag, it is transplanted to warm canopy, 15-30 DEG C of temperature, humidity 85%, one
Crop field, 99% or more survival rate can be moved on to after week.
The present invention uses vegetative manner, so that maternal merit is completely retained, and make by Shoot-tip Grafting In Vitro
Subculture is more more excellent than maternal.And the problem of woody tissue-cultured seedling quality of rooting difference is breached, tissue-cultured seedling well developed root system, completely, at
Motility rate is high, without hardening.
Description of the drawings：
Fig. 1：State after the completion of Kiwi berry root induction in embodiment 1；
Fig. 2：State after the completion of traditional tissue cultures root induction；
Fig. 3：The dimensional structure diagram of embodiment 3；
Fig. 4：The cross-sectional view of embodiment 3；
Fig. 5：The overlooking structure diagram of root bed described in embodiment 3.
Specific implementation mode
Below in conjunction with preferred embodiment, to the specific implementation mode that provides according to the present invention, details are as follows：
Embodiment 1
Using Xu Xiang Kiwi berrys as group training material, tissue cultures are carried out using the method for the present invention.
1. acquiring the fresh cane of Kiwi berry, clear water wash clean.Cane is cut into the stem section of 4cm or so, at least retains 2 armpits
Bud, 0.1% mercuric chloride sterilizing 15m, 75% alcohol sterilizing 5m, sterile water wash 3-5 times.It is inserted into MS+ sucrose 30g/L+ agar 6g/L+
In the culture medium of 6-BA0.8mg/L+IBA0.5mg/L, illumination 2000-3000Lx, 14h, 25 ± 2 DEG C of temperature, humidity 70%, training
After supporting 25 days or so, adventitious bud is born at axillary bud position, obtains primary.
2. the adventitious bud formed on callus is inserted into MS+ sucrose 30g/L+ agar 6g/L by squamous subculture (Multiplying culture)
On the culture medium of+6-BA1.2mg/L+IAA0.8mg/L, illumination 2000-3000LX, 14h, 25 ± 2 DEG C of temperature, humidity 70%, training
It supports 25 days, 3-5 times of appreciation rate.
3. root induction after proliferation 3cm or more is reached, the plant with 3 or more cotyledons is inserted into special take root in equipment
In the matrix in face, which is MS+ vermiculites, light application time 16h, 1500-2000LX, 25 ± 2 DEG C of temperature, gas concentration lwevel
950ppm, humidity 80%, 1000 grades of culturing room's cleanliness factor or more.Culture forms well developed root system Kiwi berry after 20 days is completely planted
Strain.
4. a Kiwi berry intact plant is transplanted to inside degradation bag, it is transplanted to warm canopy, 15-30 DEG C of temperature, humidity 85%, one
Crop field, 99% or more survival rate can be moved on to after week.
Embodiment 2
Using Xu Xiang Kiwi berrys as group training material, tissue cultures are carried out using different methods.
Experimental group：Use the method for the embodiment of the present invention 1；
Compare 1 group：Culture medium is used by root induction step：0.7mg·L-1IBA、30mg·L-1Sucrose, 7mg
L-1The 1/2MS culture mediums of agar, other are identical as experimental group.
Compare 2 groups：Culture medium is used by root induction step：In 1/2MS+NAA0.2mgL-1, other and experiment
Group is identical.
Compare 3 groups：Light application time 10h used by root induction step, 25 ± 2 DEG C of temperature, humidity 85%, other and reality
It is identical to test group.
Compare 4 groups：Gas concentration lwevel 1200ppm used by root induction step, other are identical as experimental group.
| It takes root pollution rate | Transplanting survival rate | It takes root situation | Whether hardening | |
| Experimental group | 0.5% | 99% | It is good | It is no |
| Compare 1 group | 5% | 86% | Generally | It is |
| Compare 2 groups | 6% | 91% | Generally | It is |
| Compare 3 groups | 1% | 95% | Preferably | It is no |
| Compare 4 groups | 1% | 93% | Generally | It is |
Embodiment 3
A kind of special equipment of taking root of tissue cultures, as shown in Figures 3 to 5, including：Box body 1, box cover 2 and root bed 3, it is described
Box cover 2 is connected by hinge 26 with box body 1, can be overturn with respect to box body 1；Described bed 3 is by several orthogonal partition board groups
At described bed 3 has the root bed region 30 for housing culture medium of several mutually independent groined types, the box cover 2
Bottom surface on be provided with LED light 20, several ventilation holes 10 are offered in the side wall surface of the box body 1, residing for the ventilation hole 10
Position is higher than the top surface of root bed 3.The root that macaque peach seedling is grown can all be separated by root bed 3, after the completion of avoiding training seedling, it
Warped roots in together, time-consuming and laborious when they are detached and easy damaged root system.
The above described is only a preferred embodiment of the present invention, being not that the invention has other forms of limitations, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
Imitate embodiment.But it is every without departing from technical solution of the present invention content, according to the technical essence of the invention to above example institute
Any simple modification, equivalent variations and the remodeling made, still fall within the protection domain of technical solution of the present invention.
Claims (3)
1. a kind of method carrying out tissue cultures using Kiwi berry stem, which is characterized in that the method for root induction is：
3cm or more is reached after proliferation, the plant with 3 or more cotyledons is inserted into the matrix inside special equipment of taking root, should
Matrix is MS+ vermiculites, light application time 16h, 1500-2000LX, 25 ± 2 DEG C, gas concentration lwevel 950ppm of temperature, humidity
80%, 1000 grades of culturing room's cleanliness factor or more；Culture forms the Kiwi berry intact plant of well developed root system after 20 days.
2. the method according to claim 1 for carrying out tissue cultures using Kiwi berry stem, which is characterized in that MS+ vermiculites are
Every liter of MS solution adds 800g vermiculites.
3. the method according to claim 1 or 2 for carrying out tissue cultures using Kiwi berry stem, which is characterized in that specific step
Suddenly it is：
1) the fresh cane of Kiwi berry, clear water wash clean are acquired.Cane is cut into the stem section of 4cm or so, at least retains 2 axillary buds,
0.1% mercuric chloride sterilizing 15m, 75% alcohol sterilizing 5m, sterile water wash 3-5 times.It is inserted into MS+ sucrose 30g/L+ agar 6g/L+6-
In the culture medium of BA0.8mg/L+IBA0.5mg/L, illumination 2000-3000Lx, 14h, 25 ± 2 DEG C of temperature, humidity 70%, culture
After 25 days or so, adventitious bud is born at axillary bud position, obtains primary.
2) adventitious bud formed on callus is inserted into MS+ sucrose 30g/L+ agar 6g/L+6- by squamous subculture (Multiplying culture)
On the culture medium of BA1.2mg/L+IAA0.8mg/L, illumination 2000-3000LX, 14h, 25 ± 2 DEG C of temperature, humidity 70%, culture
25 days, 3-5 times of appreciation rate,
3) 3cm or more is reached after proliferation, the plant with 3 or more cotyledons is inserted into inside special equipment of taking root for root induction
In matrix, which is MS+ vermiculites, light application time 16h, 1500-2000LX, 25 ± 2 DEG C of temperature, gas concentration lwevel
950ppm, humidity 80%, 1000 grades of culturing room's cleanliness factor or more.Culture forms well developed root system Kiwi berry after 20 days is completely planted
Strain；
4) Kiwi berry intact plant is transplanted to inside degradation bag, is transplanted to warm canopy, 15-30 DEG C of temperature, humidity 85%, after a week
Crop field can be moved on to.
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| CN201810083297.0A CN108271691A (en) | 2018-01-29 | 2018-01-29 | A method of carrying out tissue cultures using Kiwi berry stem |
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| CN201810083297.0A CN108271691A (en) | 2018-01-29 | 2018-01-29 | A method of carrying out tissue cultures using Kiwi berry stem |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110140658A (en) * | 2019-06-26 | 2019-08-20 | 云南师范大学 | A method of Kiwi berry seedling is cultivated using Stem Segments In Chinese Gooseberry forming layer stem cell |
| CN111713410A (en) * | 2020-07-03 | 2020-09-29 | 四川农业大学 | Kiwi explant detoxification method |
| CN114766367A (en) * | 2022-05-18 | 2022-07-22 | 西北农林科技大学 | Kiwi fruit tissue culture method |
| CN115413579A (en) * | 2022-09-20 | 2022-12-02 | 广西壮族自治区药用植物园 | Wild kiwi fruit tissue culture method and catechin preparation method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110140658A (en) * | 2019-06-26 | 2019-08-20 | 云南师范大学 | A method of Kiwi berry seedling is cultivated using Stem Segments In Chinese Gooseberry forming layer stem cell |
| CN111713410A (en) * | 2020-07-03 | 2020-09-29 | 四川农业大学 | Kiwi explant detoxification method |
| CN114766367A (en) * | 2022-05-18 | 2022-07-22 | 西北农林科技大学 | Kiwi fruit tissue culture method |
| CN115413579A (en) * | 2022-09-20 | 2022-12-02 | 广西壮族自治区药用植物园 | Wild kiwi fruit tissue culture method and catechin preparation method |
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Effective date of registration: 20200731 Address after: 722300 Baoji County, Shaanxi Province Meixian (state) kiwifruit Industrial Park testing center two floor Applicant after: BAOJI SONGLIANG AGRICULTURAL TECHNOLOGY Co.,Ltd. Applicant after: Shaanxi Pufeng Modern Agriculture Co., Ltd Address before: 722300 Baoji County, Shaanxi Province Meixian (state) kiwifruit Industrial Park testing center two floor Applicant before: BAOJI SONGLIANG AGRICULTURAL TECHNOLOGY Co.,Ltd. |
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