CN106665133A - Sterile grafting method of Arabidopsis/thellungiella halophila - Google Patents
Sterile grafting method of Arabidopsis/thellungiella halophila Download PDFInfo
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- CN106665133A CN106665133A CN201710028466.6A CN201710028466A CN106665133A CN 106665133 A CN106665133 A CN 106665133A CN 201710028466 A CN201710028466 A CN 201710028466A CN 106665133 A CN106665133 A CN 106665133A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/30—Grafting
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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
Abstract
The invention discloses a sterile grafting method of Arabidopsis/thellungiella halophila and fills up the technical blank of related grafting methods in the prior art. According to the technical scheme, the method includes: using a sterile thellungiella halophila seedling 7-10 days after germination as the stock, and using the overground part of a sterile Arabidopsis seedling 3-5 days after germination as the scion; grafting under a sterile condition, placing the grafted seedling on a 1/2MS culture medium with agar powder concentration being 1.2%, performing vertical culture under 18-22 DEG C and short-day illumination, and transferring the grafted seedling into a 1/2hogland nutrient solution for water culture or nutrient soil culture. The sterile grafting method has the advantages that the method is simple and practicable, convenient to operate, capable of reducing pollution, high in survival rate, low in damage and low in cost; the sterile grafting method for different species provides a new thought for the varietal improvement of gardening, fruits, vegetables, and the like, and a basic guarantee is provided for plant grafting in plant molecular mechanism researches.
Description
Technical field
The present invention relates to the graft technology field of small body type plant test tube seedling, more particularly to a kind of model plant arabidopsiss
And the sterile grafting method between salt tolerant model plant salt mustard.
Background technology
Grafting, is one kind of the vegetative reproduction in asexual propagation, is entered using the function after plant injury with wound healing
Capable.It has been widely used in the production of agricultural, forestry and gardening plant and scientific research and development as an agriculture production technology
In.Start so far at the beginning of the last century, particularly 70, the eighties, people have carried out numerous studies to grafting rationale.But some
Mechanism problem is not still very apparent at present.
Arabidopsiss are a kind of small-sized weeds of Cruciferae, because it has the spies such as period of duration is short, the little, genome of individuality is little
Point, for a long time by as hereditism, molecular biology, Developmental Biology research model plant.Therefore, for arabidopsiss
Research has caused the great attention of various countries scientist, and its engrafting method is also increasingly mature, with regard to arabidopsiss are from grafting and intend south
The correlational study report of grafting between mustard and mutant is continued to bring out.For example, patent " a kind of engrafting method of arabidopsiss "
(CN104604539A) engrafting method of a kind of arabidopsiss seeding-growth and Arabidopsis Mutants is disclosed, the engrafting method has side
The advantages of method is easy, simple to operate, injury is little；But the grafting of selection, grafting condition of culture sterilization method, to(for) seed etc.
Still need and further improve.
Salt mustard belongs to Cruciferae salt mustard, and using salt mustard as salt tolerant model plant Mechanism of Salt-tolerant is studied, more next
More paid attention to by scientists, but the research about salt mustard engrafting method is not reported so far.
It is well known that the factor of impact grafting survival depends primarily on the affinity of stock and scion.So-called affinity is
Refer to similarity of the stock with scion in organizational structure, heredity and physiological property, can be coadapted after grafting survival
Ability.The grafting of kindred plant, its affinity is most strong, and sibship is more remote, and its affinity is weaker.Plant between not belonging to together
Thing grafting difficulty is just bigger.Additionally, in addition to the affinity of scion and stock, stock also affects grafting with the quality of scion
The key factor for surviving, in addition, the difference of the aspect such as temperature, humidity and illumination can also affect the survival rate of grafting.And, it is different
The graft survival rate of the different times of species is also different, even same species are in different times, the survival rate of its graft
Differ greatly.
To sum up, affect grafting survival factor have a lot, arabidopsiss and salt mustard as the equal two kinds of plants not belonged to together, its
The more notable increase of grafting difficulty, and the report of arabidopsiss and salt mustard engrafting method is not also related at present, still belong to technical
It is blank.
The content of the invention
For above-mentioned the deficiencies in the prior art, it is an object of the invention to provide a kind of aseptic grafting of arabidopsiss/salt mustard
Method.The grafting for not belonging to inter-planting thing together is realized, has the advantages that method is simple, high graft survival rate.
For achieving the above object, the present invention is adopted the following technical scheme that：
A kind of sterile grafting method of arabidopsiss/salt mustard, step is as follows：
(1) stock prepares：By salt canola seed it is sterile-processed after, be seeded in the 1/2MS culture medium containing agar powder, layer
Product 1 week after proceed under long-day illumination condition cultivate, after sprouting 7-10 days as rootstock seedling；
(2) scion prepares：When salt mustard sprouts, by arabidopsiss seed disinfection, and it is seeded in containing agar powder
1/2MS culture medium on, lamination or be placed directly within long-day conditions cultivate, after sprouting 3-5 days as scion；
(3) grafting between arabidopsiss and salt mustard is carried out under aseptic condition；
(4) graft culture：The arabidopsiss for grafting/salt mustard complex is placed under the conditions of short-day and is cultivated 6-8 days；
(5) transplanting of grafting：The successful grafting transplantation of seedlings of grafting is continued into mill water culture nutrient solution or Nutrition Soil culture.
Preferably, in step (1) and step (2), using identical method to seed disinfection, specific sterilization
Method is：With NaClO that concentration is 0.5% sterilization 6-8 minutes, then with sterile water wash 4-6 time.
Preferably, in step (1) and step (2), the content of agar is 1-1.5% in the 1/2MS culture medium；Further
Preferably 1.2%.
Preferably, in step (1) and step (2), the agar in 1/2MS culture medium is Sigma agar.
Preferably, in step (1) and step (2), the long-day condition of culture is：16h illumination/8h is dark, illumination condition
Under temperature be 21-23 DEG C, the temperature under dark condition is 18-20 DEG C, and light intensity is 8000lux, and relative humidity is more than 70%.
Preferably, in step (3), the method for the grafting is：By the true leaf and growing point of the salt mustard of 7-10 days after sprouting
Cut, only retain two panels cotyledon as stock；The arabidopsiss hypocotyls of 3-5 days after sprouting are cut at upper end 1/4 as connecing
Fringe, is grafted onto on salt mustard stock.
Preferably, in step (4), the condition of short-day culture is：8h illumination/16h is dark, and the temperature under illumination condition is
21-23 DEG C, the temperature under dark condition is 18-20 DEG C, and light intensity is 8000lux, and relative humidity is more than 70%.
Preferably, in step (4), the mill water culture nutrient solution is 1/2Hogland nutritional solutions；The Nutrition Soil is by soil, trematodiasiss
Stone and perlite in mass ratio 3:2:1 makes.
In above-mentioned engrafting method, described seed disinfection, sowing and grafting procedures are operated in aseptic super-clean bench.
In above-mentioned engrafting method, in step (3), the true leaf and growing point of salt mustard are cut using double-edged razor blade.
Beneficial effects of the present invention：
First there is provided a kind of sterile grafting method of arabidopsiss/salt mustard, the present invention is with germination and growth to spy for present invention system
The timing phase, that is, the salt mustard of 7-10 days is stock after sprouting, the sterile grafting method of the arabidopsiss of 3-5 days with after sprouting as scion.
The present invention, with the single sterilizing reduction pollutions of NaClO, is conducive to normal seed germination by directly；By from 1.2% agar
With the use of double-edged razor blade, be conducive to obtaining that otch is smooth, it is smooth and injure minimum tangent plane；Transferred by operating under anatomical lens
Connect, be conducive to stock cambial with scion preferably identical, improve grafting success rate.
Can be solved due to arabidopsiss, the little operating difficultiess for bringing of salt mustard plant, otch using the engrafting method of the present invention
Out-of-flatness, graft survival rate be low and the problems such as high cost etc..The method of the present invention is simple and easy to do, low cost, efficiency high, is small-sized
Plant grafting provides new approaches, new method and new way, while dividing now in economical character, molecule mechanism etc. for plant graft
Correlation theory research in terms of son and physiology provides the foundation.
Description of the drawings
The Figure of description for constituting the part of the application is used for providing further understanding of the present application, and the application's shows
Meaning property embodiment and its illustrated for explaining the application, does not constitute the improper restriction to the application.
Fig. 1：The successful arabidopsiss of grafting/salt mustard grafting；In figure, B is the grafting part enlarged drawing of A.
Fig. 2：Arabidopsiss/salt mustard grafting procedures simulation drawing.
Fig. 3：Homemade mes holder.
Specific embodiment
It is noted that described further below is all exemplary, it is intended to provide further instruction to the application.Unless another
Indicate, all technologies used herein and scientific terminology are with usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in this manual using term "comprising" and/or " bag
Include " when, it indicates existing characteristics, step, operation, device, component and/or combinations thereof.
Term explanation：
1/2MS culture medium：Refer to the MS culture medium after a great number of elements composition in MS culture medium is halved.MS culture medium is existing
There is in technology a conventional culture medium, the formula composition of the culture medium is well known in the prior art (such as patent CN104604539A
Disclosed in MS culture medium), the culture medium can be commercially available, it is also possible to voluntarily prepare.
As background technology is introduced, the report of arabidopsiss and salt mustard engrafting method is not also related in prior art,
In order to solve technical problem as above, present applicant proposes a kind of sterile grafting method of arabidopsiss/salt mustard, specific as follows：
(1) preparation of stock and scion material：Be first by the little salt canola seed of full seed with 0.5% sodium hypochlorite
Sterilization, then sows in the 1/2MS culture medium that agar powder concentration is 1.2%, and 4 DEG C of laminations are placed in long-day illumination training after 1 week
Vertically cultivate in foster case, it is standby as rootstock seedling after sprouting 7-10 days.When salt mustard sprouts, with same sterilization method south will be intended
Canola seed carries out disinfection and plants in the 1/2MS culture medium that agar powder concentration is 1.2%, 4 DEG C of laminations or is placed directly within the long-day
Vertically cultivate in illumination box, the grafting that 3-5 days are carried out between arabidopsiss and salt mustard after Seed Germination of Arabidopsis Pumila.
(2) grafting under aseptic condition：Growing way consistent aseptic seedling salt mustard tweezers are moved to into new agar concentration 1.2%
In 1/2MS culture medium, the true leaf and growing point of salt mustard are cut with blade, only retain two panels cotyledon standby as stock.Will
Consistent the seeing under anatomical lens of growing way has the arabidopsiss hypocotyls that true leaf has just been sprouted to cut as connecing away from the place of upper end 1/4 or so
Fringe is grafted onto on preprepared salt mustard stock (simulation drawing of grafting procedures is as shown in Figure 2).
(3) culture of graft：The arabidopsiss for grafting/salt mustard complex is placed in short-day illumination box vertically
Culture 6-8 days.
(4) transplanting of grafting：Whether the grafting complex cultivated after grafting 6-8 days is being dissected into Microscopic observation graft union
Connect, the successful grafting of grafting is needed to transplant according to experiment and continues to cultivate into water planting or Nutrition Soil.Transfer during this period
If seedlings picking scion hypocotyls have lateral root to occur, cut off in time.
In above-mentioned engrafting method, for the sterilization method of seed is：Directly with 6-8 point of the NaClO sterilizations that concentration is 0.5%
Clock, then with sterile water wash 4-6 time.This kind of method is avoided and first disinfected in alcohol, then secondary sterilization is carried out with NaClO, behaviour
Make step simplification so as to reduce secondary pollution and seed will not be sterilized and affect seed to sprout and later experiments.
In above-mentioned engrafting method, adopt agar powder concentration for 1.2% 1/2MS culture medium, both ensured enough moisture profits
In normal seed germination growth, follow-up grafting of drawing materials is conducive to again.
In above-mentioned engrafting method, from the bigger salt mustard seedling for sprouting 7-10 days, its lateral root grows, so can expire
Foot its scion --- the characteristics of arabidopsiss fast growth, graft survival rate can be greatly improved and ensure that grafting late growing stage is prosperous
Contain.Because arabidopsiss growth cycle is short, with the one hand being difficult to survive compared with seedlings grafting, on the other hand easilys lead to arabidopsiss and carry
Before bloom, grafting growing way is poor, from sprout 3-5 days tenderer seedling both beneficial to improve grafting survival rate, moreover it is possible to enter
One step ensures the vigorous strong of grafting late growing stage.
In above-mentioned engrafting method, the species of culture medium and agar powder for grafting also contributes to the survival rate of grafting,
Using the present invention culture medium and agar powder grafting survival rate up to more than 95%.
In above-mentioned engrafting method, blade used is double-edged razor blade.The edge of a knife is sharp and thin, is conducive to tangent plane smooth, to the greatest extent may be used
Can few injury cambial cell to improve the survival rate of grafting, in order to reduce pollution, present invention bamboo let has been made by oneself simply
Mes holder (see Fig. 3), and the agar concentration hardness of the 1.2% of the present invention not only can guarantee that enough moisture while also be enough to
Meet the flat smooth of facet.
In above-mentioned engrafting method, the graft after grafting is placed under short-day and cultivates, and not only contributes to improve graft
Survival rate, and it is strong to also help grafting late growing stage.This is because arabidopsiss growth cycle is short, come to harm at it
Afterwards, its growth cycle is just further greatly shortened, if under long-day conditions, may result in grafting small and weak and too early
Yield positive results and complete its life cycle.And short-day can extend its nutrient growth, so as to make up the deficiency under the long-day.
In above-mentioned engrafting method, seed disinfection is inoculated with and is operated in aseptic super-clean bench, and grafting procedures are also aseptic ultra-clean
Operate in platform, and the docking of grafting wound is operated under anatomical lens, is more beneficial for scion and the cambial docking of stock, can
To improve the success rate of grafting.
In order that those skilled in the art can clearly understand the technical scheme of the application, below with reference to tool
The embodiment of body describes the technical scheme of the application in detail.
Test material used is the conventional test material in this area in the embodiment of the present invention, can pass through commercial channel
It is commercially available.
Embodiment 1：The sterile grafting method of arabidopsiss/salt mustard
Comprise the following steps that：
(1) preparation of stock and scion material：First it is by the little salt mustard of the Shandong Province of China Dongying of full seed
(Thellungiella halophila) seed is with the hypochlorite disinfectant of 0.5% (mass concentration) and sows in agar powder
(Sigma agar) concentration (" concentration " herein is the mass fraction of agar in culture medium, similarly hereinafter) is 1.2% 1/2MS cultures
On base (domestic culture medium), this process is operated on aseptic super-clean bench.4 DEG C of laminations are placed in long-day illumination box after 1 week
Vertically culture is sprouted standby.With the sterilization method same with salt mustard by Colombia type arabidopsiss (Arabidopsis
Thaliana) seed carries out disinfection and is seeded in 1/2MS culture medium (the domestic training that agar powder (Sigma agar) concentration is 1.2%
Foster base) on, this process is operated on aseptic super-clean bench.4 DEG C of laminations or be placed directly within long-day illumination box are vertically cultivated,
The grafting that 3-5 days are carried out between arabidopsiss and salt mustard after Seed Germination of Arabidopsis Pumila.Method for seed disinfection is directly to use
Concentration is 0.5% NaClO sterilization 6-8 minutes, then with sterile water wash 4-6 time.This kind of method is avoided and first disappeared with ethanol
Poison, then secondary sterilization is carried out with NaClO, operating procedure simplification so as to reduce secondary pollution and seed will not be sterilized and affect
Seed is sprouted and later experiments.Simultaneously from the 1/2MS culture medium that agar powder concentration is 1.2%, enough moisture profits were both ensured
In normal seed germination growth, follow-up grafting of drawing materials is conducive to again.Secondly from the bigger salt mustard seedling for sprouting 7-10 days,
Its lateral root grows, can so meet its scion --- the characteristics of arabidopsiss fast growth, grafting survival can be greatly improved
Rate and guarantee grafting late growing stage are vigorous.Due to arabidopsiss growth cycle it is short, on the one hand with being difficult to survive compared with seedlings grafting, separately
On the one hand arabidopsiss Blooming is easily lead to, grafting growing way is poor, is both beneficial to carry from the tenderer seedling for sprouting 3-5 days
The survival rate of high grafting, moreover it is possible to be further ensured that the vigorous strong of grafting late growing stage.
(2) cutting and aseptic condition under grafting：The 7-10 days consistent aseptic seedling salt mustard of growing way after sprouting are moved to tweezers
In the 1/2MS culture medium (domestic culture medium) of new agar (Sigma agar) concentration 1.2%, with double-edged razor blade by the true of salt mustard
Leaf and growing point cut, and only retain two panels cotyledon standby as stock.By growing way it is consistent can see there is true leaf under anatomical lens
The arabidopsiss hypocotyls for just having sprouted are cut away from the place of upper end 1/4 or so and be grafted onto as scion on preprepared salt mustard stock,
And under anatomical lens by the two otch to connecting, and culture dish sealed with moisturizing in time.This process is grasped on aseptic super-clean bench
Make.
The double-edged razor blade edge of a knife used is sharp and thin, is conducive to tangent plane smooth, and as few as possible injury cambial cell is carrying
The survival rate of high grafting, in order to reduce pollution, we have made simple mes holder (see accompanying drawing 2) by oneself with bamboo let, and we use
1.2% agar concentration hardness not only can guarantee that enough moisture while being also sufficient for the flat smooth of facet.Due to
Stock and scion are aseptic seedling, and comparison is tender and lovely, and opening culture dish will soon wilt, and otch also easy dehydration wilting, therefore
Technical staff's action is fast in grafting procedures, and the wind speed of super-clean bench reaches minimum, and grafting finishes rapid sealing, but action is light,
Prevent graft from misplacing.
(3) culture of graft：The arabidopsiss for grafting/salt mustard complex is placed in short-day illumination box vertically
Culture 6-8 days.
(4) transplanting of grafting：Whether the grafting complex cultivated after grafting 6-8 days is being dissected into Microscopic observation graft union
Connect, the successful grafting of grafting is needed to transplant according to experiment and continues to cultivate into water planting or Nutrition Soil.For water planting
Nutritional solution is 1/2Hogland nutritional solutions, and Nutrition Soil is soil：Vermiculitum：Perlitic ratio is 3:2:1.During this period grafting connects
If fringe hypocotyls have lateral root to occur, cut off in time.
In above-mentioned engrafting method, long-day conditions are that 16h illumination/8h is dark, and the temperature under illumination condition is 21-23 DEG C,
Temperature under dark condition is 18-20 DEG C, and light intensity is 8000lux, and relative humidity is more than 70%.And it is short in above-mentioned steps 3 and 4
Sunshine condition is that 8h illumination/16h is dark, the same long-day conditions of temperature, light intensity and humidity.Graft after grafting is placed in short-day
Lower culture, not only contributes to improve the survival rate of graft, and it is strong to also help grafting late growing stage.This is because intending
Southern mustard growth cycle is short, and after it comes to harm, its growth cycle is just further greatly shortened, if in long-day conditions
Under, may result in small and weak and too early the yielding positive results of grafting and complete its life cycle.And short-day can extend its nutrient growth,
So as to make up the deficiency under the long-day.
Comparative example 1：
The embodiment 1 of selection in to(for) rootstock seedling is adjusted to respectively：After sprouting the salt mustard Seedling of 3-5 days and sprout after 11-
The salt mustard Seedling of 15 days, other operations are with embodiment 1.
Comparative example 2：
The embodiment 1 of selection in to(for) scion seedling is adjusted to：The arabidopsiss Seedling of 6-8 days after sprouting；By the choosing of rootstock seedling
Select and be adjusted to respectively：The salt mustard Seedling of 3-5 days after sprouting, sprout after 7-10 days salt mustard Seedling and the salt mustard Seedling of 11-15 days after sprouting,
Other operations are with embodiment 1.
The survival rate and later stage growing way of grafting after embodiment 1, comparative example 1 and the grafting of comparative example 2 are investigated, as a result such as table
Shown in 1.
Table 1：Impact of the sprout time length to grafting
As can be seen from Table 1, as the salt mustard and Arabidopsis thaliana Seedlings of stock and scion, the selection of its sprout time can affect
The survival rate and the growing way in grafting later stage of grafting.Compare with comparative example 1 with comparative example 2, the embodiment of the present invention 1 is from sprouting
The bigger salt mustard seedling of 7-10 days, its lateral root grows, the characteristics of can so meet its scion arabidopsiss fast growth,
Graft survival rate can be greatly improved and ensure that grafting late growing stage is vigorous.Due to arabidopsiss growth cycle it is short, with transferring compared with seedlings
Connect and on the one hand be difficult to survive, on the other hand easily lead to arabidopsiss Blooming, grafting growing way is poor, from sprouting 3-5
It tenderer seedling is both beneficial to the survival rate for improving grafting, moreover it is possible to be further ensured that the vigorous strong of grafting late growing stage.
Comparative example 3：
1/2MS culture medium containing agar in embodiment 1 is adjusted, agar is replaced with into domestic agar, will be cultivated
Base replaces with autogamy culture medium, domestic culture medium and Sigma culture medium and (is 1/2MS culture medium, difference is coming for culture medium
Source is different), and be applied in combination, remaining operation is with embodiment 1.
The survival rate of grafting after embodiment 1 and the grafting of comparative example 3 is investigated, as a result as shown in table 2.
Table 2：The impact of different agar powders (1.2%) and 1/2MS culture medium to grafting success rate
As can be seen from Table 2, for grafting culture medium and the species of agar powder also contributes to the survival rate of grafting, phase
There is larger difference is affected on the survival rate of grafting with the different agar powders under concentration, wherein Sigma agar powders are more beneficial for
Grafting survives.
The preferred embodiment of the application is the foregoing is only, the application is not limited to, for the skill of this area
For art personnel, the application can have various modifications and variations.It is all within spirit herein and principle, made any repair
Change, equivalent, improvement etc., should be included within the protection domain of the application.
Claims (10)
1. the sterile grafting method of a kind of arabidopsiss/salt mustard, it is characterised in that step is as follows：
(1) stock prepares：By salt canola seed it is sterile-processed after, be seeded in the 1/2MS culture medium containing agar powder, lamination 1
Proceed to after week under long-day illumination condition cultivate, after sprouting 7-10 days as rootstock seedling；
(2) scion prepares：When salt mustard sprouts, by arabidopsiss seed disinfection, and containing agar powder 1/ is seeded in
In 2MS culture medium, lamination or be placed directly within long-day conditions cultivate, after sprouting 3-5 days as scion；
(3) grafting between arabidopsiss and salt mustard is carried out under aseptic condition；
(4) graft culture：The arabidopsiss for grafting/salt mustard complex is placed under the conditions of short-day and is cultivated 6-8 days；
(5) transplanting of grafting：The successful grafting transplantation of seedlings of grafting is continued into mill water culture nutrient solution or Nutrition Soil culture.
2. sterile grafting method according to claim 1, it is characterised in that in step (1) and step (2), using identical
Method to seed disinfection, specific sterilization method is：Sterilized 6-8 minutes with the NaClO that concentration is 0.5%, so
Afterwards with sterile water wash 4-6 time.
3. sterile grafting method according to claim 1, it is characterised in that in step (1) and step (2), the 1/2MS
The content of agar is 1-1.5% in culture medium；Preferably 1.2%.
4. the sterile grafting method according to claim 1 or 3, it is characterised in that the agar in 1/2MS culture medium is
Sigma agar.
5. sterile grafting method according to claim 1, it is characterised in that in step (1) and step (2), long-day training
Foster condition is：16h illumination/8h is dark, and the temperature under illumination condition is 21-23 DEG C, and the temperature under dark condition is 18-20
DEG C, light intensity is 8000lux, and relative humidity is more than 70%.
6. sterile grafting method according to claim 1, it is characterised in that in step (3), the method for the grafting is：
The true leaf and growing point of the salt mustard of 7-10 days after sprouting are cut, only retains two panels cotyledon as stock；By 3-5 days after sprouting
Arabidopsiss hypocotyls are cut as scion at upper end 1/4, are grafted onto on salt mustard stock.
7. sterile grafting method according to claim 1, it is characterised in that in step (4), the condition of short-day culture
For：8h illumination/16h is dark, and the temperature under illumination condition is 21-23 DEG C, and the temperature under dark condition is 18-20 DEG C, and light intensity is
8000lux, relative humidity is more than 70%.
8. sterile grafting method according to claim 1, it is characterised in that in step (4), the mill water culture nutrient solution is 1/
2Hogland nutritional solutions；The Nutrition Soil is by soil, Vermiculitum and perlite in mass ratio 3:2:1 makes.
9. sterile grafting method according to claim 1, it is characterised in that described seed disinfection, sowing and grafting
Cheng Jun is operated in aseptic super-clean bench.
10. sterile grafting method according to claim 1, it is characterised in that in step (3), cut using double-edged razor blade
The true leaf and growing point of salt mustard.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108713406A (en) * | 2018-06-07 | 2018-10-30 | 齐鲁师范学院 | The engrafting method of arabidopsis and its application in restoring Arabidopsis Mutants fertility |
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