CN105230485B - A kind of method of transformation of soybean adventitious bud rooting - Google Patents
A kind of method of transformation of soybean adventitious bud rooting Download PDFInfo
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Abstract
The invention provides a kind of method of transformation of soybean adventitious bud rooting, the method specific steps taken root include：The lower end of transformation of soybean adventitious bud is immersed in taking root liquid, in the pot for growing seedlings that the transformation of soybean adventitious bud implantation after immersion is not closed, overlay film, illumination cultivation is carried out, after young leaves is grown, takes off film, continue culture to strong sprout.The taking root liquid used in the method for transformation of soybean adventitious bud rooting provided by the invention, component are simple and convenient to operate, and before adventitious bud rooting or are transplanted to after culture of rootage matrix and are not easy to grow Agrobacterium, and rooting rate can reach 96.24%.This method uses (open environment) directly in the pot for growing seedlings do not closed to take root, improve quality of rooting, cultivate and obtain healthy and strong regrowth, so as to improve the transplanting survival rate of regenerable soybean seedling, survival rate can reach 100%, final to improve transformation of soybean success rate.
Description
Technical field
The present invention relates to biological technical field, in particular to a kind of method of transformation of soybean adventitious bud rooting.
Background technology
Soybean is generally called soya bean, pulse family Glycine annual herb.China is originated in, there is cultivation all over China, is also planted extensively
Train in all over the world.Soybean is one of important cereal crops of China, and Northeast China is main producing region, and it is abundant to be that its a kind of seed contains
The crop of phytoprotein, soybean is the most frequently used to be made various bean product, squeezes soya-bean oil, makes soy sauce and extract protein.Bean dregs
Or wear into the soybean of coarse powder and be also commonly used for poultry and livestock feed.
In the 1980s, Monsanto Company researcher clones from petunia obtains resistant gene (EPsPs bases
Cause), and led the EPsPs genes of petunia in people's soybean genome with agrobacterium-mediated transformation, and then it is big to cultivate resistance glyphosate
Beans kind.1994, the resistance glyphosate genetically engineered soybean that Monsanto Company cultivates obtained american commerceization plantation license first.Turn
Transgenic soybean since popularization, be in first of all kinds of genetically modified crops always by cultivated area, is earliest commercialization, popularization in the world
The most fast genetically modified crops of application speed.
Soybean transgene will pass through channel genes, conversion adventitious bud inducing, Elongation of adventitious bud, adventitious bud rooting, plant and move
Cultivation, which grows to bloom to bear pods, obtains transgenic progeny.Survival rate after the taking root and transplant of transformation of soybean adventitious bud is to influence soybean
One of successful key link of transgenosis.In published plant transgenic technology, transformation of soybean adventitious bud rooting to plant
Maturation typically passes through following steps：(1) adventitious bud life is cut to be transferred in root media and carries out culture of rootage；(2) from training
Support in bottle and take out the soybean regrowth taken root, clean the culture medium on root, (3) are further transplanted in matrix；Continuing cultivation makes
It is healthy and strong that seedling occurs for young leaves；(4) and then it is transplanted in soil, normal management is until bearing pods.Among these, rooting induction is for shape
It is most important into complete plant, be sprout can autonomous growth committed step, pass through the autonomous nutrient absorption of regeneration plant
The nutrient supply absorbed with root hair, sprout can healthily grow, grow up to healthy and strong plant.
Commonly used root media is using MS or B5 as minimal medium in current above-mentioned rooting technique, wherein individually adding
Add indolebutyric acid (IBA) 0.2-1.0mg/L, or individually add methyl α-naphthyl acetate (NAA) 0.37-1.0mg/L.
On the research of the rooting technique of transformation of soybean adventitious bud, mainly there is " the soybean transgene that Zhu Mulan et al. is invented
Strain mixes rooting method " (application number：CN201310222316.0), " one kind solution transformation of soybean of Yang Yuesheng et al. inventions
Indefinite budling is difficult to the sterile grafting method taken root " (application number：CN201210368585.3) and one kind for bravely inventing of ginger state is big
Beans tissue cultures break up the rooting method (application number of seedling：CN201410349357.0), it is required in above rooting technique close
Close on the aseptic culture medium in blake bottle and take root.In the prior art, it is necessary to prepare solid root media, technical sophistication and big
Beans adventitious bud is transferred to easy bacterial infection in culture medium and follow-up incubation, the culture environment of solid root media
It is the physiological metabolism for being unfavorable for ratoon growth in closed blake bottle, suppresses the root of hair of regrowth, and take root in solid
The adventitious root cultivated in culture medium is of poor quality and do not have root hair compared with normal root system, and absorption function is poor.The closed loop in bottle
The rooted seedling survival ability cultivated under border is weak, is typically passed through the process tamed outside bottle, is then colonized again, in this process
In, there are some rooted seedlings dead because being not suitable with external environment condition, cause transplanting survival rate to reduce.
In view of this, it is special to propose the present invention.
The content of the invention
The purpose of the object of the invention first is to provide a kind of method of transformation of soybean adventitious bud rooting, and this method has operation
Simply, the advantages that improving the survival rate of quality of rooting, the rooting rate for improving soybean transformation adventitious bud and regenerable soybean seedling.Together
When, the container environment for avoiding sealing suppresses and to ratoon growth and the unfavorable shadow of physiological metabolism to the root of hair of regrowth
Ring, avoid regrowth microbiological contamination in rooting process, cultivate the regrowth of stalwartness.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of method of transformation of soybean adventitious bud rooting, comprises the following steps：The lower end of transformation of soybean adventitious bud is soaked
In taking root liquid, in the pot for growing seedlings for then not closing the transformation of soybean adventitious bud implantation after immersion, overlay film, illumination training is carried out
Support, after young leaves is grown, take off film, continue culture to strong plantlets and rootage.
The method of transformation of soybean adventitious bud rooting provided by the invention, can be effectively facilitated adventitious root.Simultaneously can
To suppress the growth of the microbiological contamination in follow-up rooting process, especially Agrobacterium.Can be taken root training instead of solid of the prior art
Base is supported, solid root media manufacturing process is cumbersome, technical sophistication, and whole process requires sterile working, also, soybean adventitious bud
It is transferred to easy microbiological contamination during culture medium and follow-up cultivation, the soybean especially converted in agriculture bacillus mediated method, shape
Into regenerated adventitious bud carry Agrobacterium in itself, adventitious bud is before taking root or is transferred to after root media and easily grows
Agrobacterium, the adventitious bud after microbiological contamination can not take root and death.In this method, using (open directly in the pot for growing seedlings do not closed
Environment) take root, avoid the container environment of sealing and the root of hair of regrowth suppressed and to ratoon growth and physiological metabolism
Adverse effect.Solid root media manufacturing process is cumbersome, technical sophistication, and whole process requires sterile working.Taking root liquid can
To replace solid root media of the prior art before adventitious bud is implanted into and taken root in pot for growing seedlings, come provide take root it is required
Nutrition and environment of taking root, seedling medium breathe freely it is loose be unfavorable for bacteria breed, therefore avoid in regrowth rooting process
Microbiological contamination.Solid root media quality consolidation is airtight, and the adventitious root grown wherein does not have root hair, and is educated in pot for growing seedlings
For seedling matrix close to soil environment, the root grown possesses abundant root hair, and root hair plays important absorption.Therefore the soybean
The method of conversion adventitious bud rooting improves quality of rooting, cultivates and obtains healthy and strong regrowth, so as to improve transformation of soybean adventitious bud
Rooting rate, the transplanting survival rate of regenerable soybean seedling, it is final to improve transformation of soybean success rate.
Preferably, described method specifically includes following steps：
(1) in the taking root liquid for being immersed in the lower end of transformation of soybean adventitious bud；
(2) seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the soybean soaked in step (1)
Conversion adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；
(3) the transformation of soybean adventitious bud in step (2) is subjected to illumination cultivation, the thin of covering is opened after young leaves is grown
Film, continue to cultivate the transformation of soybean adventitious shoot regeneration seedling, until regrowth is healthy and strong.
Contribute to transformation of soybean adventitious bud rooting by taking root liquid immersion, the pot for growing seedlings do not closed is that taking root for adventitious bud carries
For open growing environment, adventitious bud is set to take root naturally, unfettered, the root for obtaining healthy stalwartness is transformation of soybean adventitious bud
The guarantee that transplanting survival rate and big bean seedlings survive.Watering and overlay film provide and kept moisture needed for transformation of soybean adventitious bud rooting.
Energy necessary to illumination provides photosynthesis for transformation of soybean adventitious bud.Using this method, transformation of soybean adventitious bud is taken root
Rate can reach 96.24%, and transplanting survival rate reaches 100%.
Preferably, in the taking root liquid, the component of following concentration is included：Indolebutyric acid 0.5-2.0mg/L, methyl α-naphthyl acetate 0.2-
0.5mg/L, cephalosporin 200-300mg/L, pH=5.5-6.0.
Preferably, in the taking root liquid, the component of following concentration is included：Indolebutyric acid 0.5-1.5mg/L, methyl α-naphthyl acetate 0.2-
0.5mg/L, cephalosporin 240-260mg/L, pH=5.7-5.9.
Indolebutyric acid in the taking root liquid is a kind of good root-growing agent, is mainly used in rooting of cuttings, can induce root substance
Formation, promote cell differentiation and division, be advantageous to new root generation and the differentiation of fibrovascular system, promote the shape of cutting adventitious root
Into still, indolebutyric acid is not easy to transport in vivo after being absorbed by plants, and often rests on applied position, is primarily to facilitate
Rooting of cuttings, the root born is carefully dredged, branch is more；Methyl α-naphthyl acetate is a kind of auximone, available for Plant Tissue Breeding.Its
In, methyl α-naphthyl acetate helps lend some impetus to the generation of main root, and facilitation, two kinds of growths are played in formation of the indolebutyric acid for capillary root
During plain independent role, effect and unobvious, positive effect during collective effect, rooting rate can reach 96.24%.Two kinds of auxin
It can be stabilized under weakly acidic condition.Cephalosporin is a kind of conventional antibiotic, can suppress Agrobacterium and other are thin
The growth of bacterium.Because auxin release contained in solid root media is slow, spy is substituted using taking root liquid processing adventitious bud
Solid root media, adventitious bud can be made to absorb active ingredient rapidly, and simplify operating process, improve soybean transformation adventitious bud
Rooting rate.
Preferably, the time of the immersion is 5-30 minutes.
Sufficient soak time ensure that transformation of soybean adventitious bud fully absorbs the auxin in taking root liquid, promote main root and
The growth of capillary root.
Preferably, the time of the illumination cultivation is 14-16 hours/day, and the intensity of illumination cultivation is 3000-10000LX,
The temperature of illumination cultivation is 22-25 DEG C.
Ensure sufficient illumination, promote adventitious bud photosynthesis, necessary material is provided for taking root for adventitious bud.
Preferably, after culture is continued, in addition to：Remove pot for growing seedlings, by the adventitious bud taken root together with seedling medium one
Transplant planting is played to soil or to be transferred in big culture vessel.
Treat that transformation of soybean adventitious bud bears the root of health, it is necessary to be transplanted to bigger culture vessel together with seedling medium
Or in soil, contribute to big bean seedlings to grow up, until the ripe acquisition seed that bears pods.
Preferably, the length of described transformation of soybean adventitious bud is more than 3cm.
The adventitious bud that length is more than 3cm is cut, preferable adventitious bud is advantageous to transformation of soybean adventitious bud rooting, improves soybean
Convert the rooting rate and transplanting survival rate of adventitious bud.
Preferably, the quantity of the young leaves is 2-3 pieces.
Overlay film is raised when 2-3 piece young leaves is grown, big bean seedlings is adapted to external environment, improves soybean shoot survival percent.
Preferably, it is described to continue also to include watering step during culture.
Carried out after overlay film is raised, big bean seedlings are watered, ensure moisture necessary to big bean seedlings continued growth.
Compared with prior art, beneficial effects of the present invention are：
(1) method of transformation of soybean adventitious bud rooting provided by the invention, can significantly improve transformation of soybean adventitious bud
Rooting rate, rooting rate reach 96.24%；
(2) taking root liquid used in the method for transformation of soybean adventitious bud rooting provided by the invention has formula composition letter
It is single, easy to operate, improve quality of rooting the advantages of；
(3) method of transformation of soybean adventitious bud rooting provided by the invention, can significantly improve transformation of soybean adventitious bud
Root system quality, improve transplanting survival rate, survival rate to 100%.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, it is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
The method of this transformation of soybean adventitious bud rooting provided in an embodiment of the present invention specifically includes following steps：
The lower end of transformation of soybean adventitious bud is immersed in taking root liquid, is then implanted into the transformation of soybean adventitious bud after immersion
In the pot for growing seedlings do not closed, overlay film, illumination cultivation is carried out, after young leaves is grown, takes off film, continue culture to well developed root system, regeneration
Seedling is healthy and strong.
Embodiment 2
The method of this transformation of soybean adventitious bud rooting provided in an embodiment of the present invention specifically includes following steps：
S21, by length be more than 3cm transformation of soybean adventitious buds lower end immersion taking root liquid in, soak 5 minutes；
Wherein, the composition of taking root liquid is as follows：Indolebutyric acid 0.5mg/L, methyl α-naphthyl acetate 0.5mg/L, cephalosporin 200mg/L,
PH=5.5；
S22, seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the soybean soaked in step S21
Conversion adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；
S23, the transformation of soybean adventitious bud in step S22 carries out to illumination cultivation, time of illumination cultivation for 14 hours/
My god, the intensity of illumination cultivation is 3000LX, and the temperature of illumination cultivation is 22 DEG C, and the film of covering is opened after 2 young leaves are grown,
Continue to cultivate the soybean regrowth, until regrowth is healthy and strong；
S24, remove pot for growing seedlings, the adventitious bud taken root together with seedling medium transplant planting to soil or is transferred to big
Culture vessel in.
Embodiment 3
The method of this transformation of soybean adventitious bud rooting provided in an embodiment of the present invention specifically includes following steps：
S31, by length be more than 3cm transformation of soybean adventitious buds lower end be immersed in taking root liquid, soak 20 minutes；
Wherein, the composition of taking root liquid is as follows：Indolebutyric acid 1.0mg/L, methyl α-naphthyl acetate 0.2mg/L, cephalosporin 300mg/L,
PH=6.0.
S32, seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the soybean soaked in step S31
Conversion adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；
S33, the transformation of soybean adventitious bud in step S32 carries out to illumination cultivation, time of illumination cultivation for 15 hours/
My god, the intensity of illumination cultivation is 6000LX, and the temperature of illumination cultivation is 23 DEG C, and the film of covering is opened after 3 young leaves are grown,
Continue to cultivate the soybean regrowth, water, until regrowth is healthy and strong；
S34, remove pot for growing seedlings, the adventitious bud taken root together with seedling medium transplant planting to soil or is transferred to big
Culture vessel in.
Embodiment 4
The method of this transformation of soybean adventitious bud rooting provided in an embodiment of the present invention specifically includes following steps：
S41, by length be more than 3cm transformation of soybean adventitious buds lower end be immersed in taking root liquid, soak 30 minutes；
Wherein, the composition of taking root liquid is as follows：Indolebutyric acid 1.0mg/L, methyl α-naphthyl acetate 0.5mg/L, cephalosporin 250mg/L,
PH=5.8；
S42, seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the soybean soaked in step S41
Conversion adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；
S43, the transformation of soybean adventitious bud in step S42 carries out to illumination cultivation, time of illumination cultivation for 15 hours/
My god, the intensity of illumination cultivation is 6000LX, and the temperature of illumination cultivation is 23 DEG C, and the film of covering is opened after 3 young leaves are grown,
Continue to cultivate the soybean regrowth, water, until regrowth is healthy and strong；
S44, remove pot for growing seedlings, the adventitious bud taken root together with seedling medium transplant planting to soil or is transferred to big
Culture vessel in；
Embodiment 5
The method of this transformation of soybean adventitious bud rooting provided in an embodiment of the present invention specifically includes following steps：
S51, by length be more than 3cm transformation of soybean adventitious buds lower end be immersed in taking root liquid, soak 30 minutes；
Wherein, the composition of taking root liquid is as follows：Indolebutyric acid 1.5mg/L, methyl α-naphthyl acetate 0.2mg/L, cephalosporin 250mg/L,
PH=5.8；
S52, seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the soybean soaked in step S51
Conversion adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；
S53, the transformation of soybean adventitious bud in step S52 carries out to illumination cultivation, time of illumination cultivation for 16 hours/
My god, the intensity of illumination cultivation is 10000LX, and the temperature of illumination cultivation is 25 DEG C, and the thin of covering is opened after 3 young leaves are grown
Film, continue to cultivate the soybean regrowth, water, until regrowth is healthy and strong；
S54, remove pot for growing seedlings, the adventitious bud taken root together with seedling medium transplant planting to soil or is transferred to big
Culture vessel in.
Embodiment 6
The method of this transformation of soybean adventitious bud rooting provided in an embodiment of the present invention specifically includes following steps：
S61, the lower end of transformation of soybean adventitious bud is immersed in taking root liquid, soaked 25 minutes；
Wherein, the composition of taking root liquid is as follows：Indolebutyric acid 2.0mg/L, methyl α-naphthyl acetate 0.35mg/L, cephalosporin 250mg/L,
PH=5.8；
S62, seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the soybean soaked in step S61
Conversion adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；
S63, the transformation of soybean adventitious bud in step S62 carries out to illumination cultivation, time of illumination cultivation for 16 hours/
My god, the intensity of illumination cultivation is 5000LX, and the temperature of illumination cultivation is 25 DEG C, and the film of covering is opened after 3 young leaves are grown,
Continue to cultivate the soybean regrowth, until regrowth is healthy and strong.
Wherein, in embodiment 1-6, the method for obtaining transformation of soybean adventitious bud specifically includes following steps：
A, disease-free, full ripe soya seeds are selected first to be carried out disinfection 16h with chlorine method, are inoculated into germination medium
On, 5d is sprouted, obtains aseptic seedling；
Wherein, the composition of germination medium is：B5,0.5mg/L 6-BA, 3% sucrose, 0.6% agar, pH=5.8；
B, prepared by explant：Cutting cotyledonary node, takes out aseptic seedling on superclean bench, cuts cotyledon, retains under 2-3mm
Plumular axis, rip cutting separation two panels cotyledon, removes terminal bud, lateral bud, is longitudinally rowed dry at cotyledonary node several knives with point of a knife, obtained outside cotyledonary node
Implant, soak the good soybean cotyledon node 60min of cutting with fluid nutrient medium；
Wherein, the composition of fluid nutrient medium is：1/10B5,1.7mg/L 6-BA, 200 μm of ol/ acetosyringones, 400mg/
L Cys, 1mmol/L dithiothreitol (DTT)s, 1mmol/L sodium thiosulfate, 3% sucrose, pH=6.6, modulation Agrobacterium are dense
Degree OD600 reaches 0.5, and Agrobacterium receives mycin selectable plant marker containing card；
C, soybean cotyledon node is transferred to and co-cultured on base, under 24 DEG C, 12h illumination/12h dark conditions, co-culture 7
My god；
Wherein, the composition for co-culturing base is：1/10B5,1.7mg/L 6-BA, 200 μm of ol/L acetosyringones, 400mg/L
Cys, 1mmol/L dithiothreitol (DTT)s, 1mmol/L sodium thiosulfate, 3% sucrose, 0.8% agar, pH=6.6；
D, after the soybean cotyledon node after above-mentioned co-cultivation being carried out into de- bacterium, go to adventitious bud inducing and screening and culturing medium is enterprising
Row bud induce and screening and culturing, 15 days subcultures once, until adventitious bud is grown；
Wherein, induction and screening and culturing based component are：B5、1.7mg/L 6-BA、100mg/L Km、250mg/L Carb、
250mg/L Cef, 3% sucrose, 0.8% agar, pH=5.8；
E, above-mentioned adventitious bud is transferred in bud elongation medium, obtains transformation of soybean adventitious bud；
Wherein, bud elongation medium composition is：B5、1.0mg/L GA3、100mg/L Km、250mg/L Carb、250mg/
L Cef, 3% sucrose, 0.8% agar, pH=5.8.
Influence of the auxin concentration to rooting rate in the present invention of test example 1
This test example is using single addition indolebutyric acid and methyl α-naphthyl acetate as control, and measure different component is to transformation of soybean adventitious bud
The influence of rooting rate.
In taking root liquid of the present invention, independent addition indolebutyric acid is between 0.5-2.0mg/L with methyl α-naphthyl acetate in 0.2-0.5mg/L
Between, its rooting rate is less than 90%, and two kinds of auxin are used cooperatively, the rooting rate of transformation of soybean adventitious bud 90% with
On, wherein optimal with the component arranging effect in embodiment 5, its rooting rate reaches highest, reaches 96.24%.
Influence of the culture environment to rooting rate and transplanting survival rate in the present invention of test example 2
This test example is control with solid root media closing culture, tests transformation of soybean adventitious bud under varying environment
Rooting rate and transplanting survival rate.
Wherein, the composition of solid root media is：50%MS, 1.5mg/L IBA, 0.2mg/L NAA, 250mg/L
Cef, 3% sucrose, 0.8% agar, pH=5.8.
The method of transformation of soybean adventitious bud rooting provided by the invention, seedling medium is implanted directly into after being handled with taking root liquid
In.Rooting rate reaches more than 90%, can reach 96.24%, transplanting survival rate reaches 100%, respectively than in culture medium
In take root and improve 56.05% and 46.64%.
In summary, the taking root liquid that the method for transformation of soybean adventitious bud rooting provided by the invention uses, component is simple, behaviour
Facilitate, and before adventitious bud rooting or be transplanted to after culture of rootage matrix and be not easy to grow Agrobacterium, can replace
Solid root media of the prior art.Indolebutyric acid, methyl α-naphthyl acetate in taking root liquid and cephalosporin collocation use, can be with
Simplify operating process, improve the rooting rate of transformation of soybean adventitious bud, rooting rate can reach 96.24%.It is provided by the invention big
Beans convert adventitious bud rooting method, using directly in the pot for growing seedlings do not closed (open environment) take root, avoid sealing
Container environment the root of hair of regrowth is suppressed and to ratoon growth and the adverse effect of physiological metabolism, while avoid again
Microbiological contamination during raw seedling rooting, improves quality of rooting, cultivates and obtains healthy and strong regrowth, so as to improve the transplanting of regenerable soybean seedling
Survival rate, survival rate can reach 100%, final to improve transformation of soybean success rate.
Although illustrate and describing the present invention with specific embodiment, but will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (7)
- A kind of 1. method of transformation of soybean adventitious bud rooting, it is characterised in that described method specifically includes following steps：(1) lower end of transformation of soybean adventitious bud is immersed in the taking root liquid of transformation of soybean adventitious bud；(2) seedling medium is fitted into the pot for growing seedlings do not closed, watered, then by the transformation of soybean soaked in step (1) Adventitious bud is implanted into the pot for growing seedlings do not closed equipped with seedling medium, is covered with film；(3) the transformation of soybean adventitious bud in step (2) is subjected to illumination cultivation, the film of covering is opened after young leaves is grown, after It is continuous to cultivate the transformation of soybean adventitious shoot regeneration seedling, until regrowth is healthy and strong；The length of transformation of soybean adventitious bud described in step (1) is more than 3cm；In the taking root liquid, include the component of following concentration：Indolebutyric acid 0.5-2.0mg/L, methyl α-naphthyl acetate 0.2-0.5mg/L, head P0-357 200-300mg/L, pH=5.5-6.0.
- 2. the method for transformation of soybean adventitious bud rooting according to claim 1, it is characterised in that in the taking root liquid, bag Include the component of following concentration：Indolebutyric acid 0.5-1.5mg/L, methyl α-naphthyl acetate 0.2-0.5mg/L, cephalosporin 240-260mg/L, pH =5.7-5.9.
- 3. the method for transformation of soybean adventitious bud rooting according to claim 1, it is characterised in that the time of the immersion is 5-30 minutes.
- 4. the method for transformation of soybean adventitious bud rooting according to claim 1, it is characterised in that the illumination cultivation when Between be 14-16 hours/day, the intensity of illumination cultivation is 3000-10000LX, and the temperature of illumination cultivation is 22-25 DEG C.
- 5. the method for transformation of soybean adventitious bud rooting according to claim 1, it is characterised in that after culture is continued, Also include：Remove pot for growing seedlings, the adventitious bud taken root together with seedling medium transplant planting to soil or is transferred to big training Support in container.
- 6. the method for transformation of soybean adventitious bud rooting according to claim 1, it is characterised in that described in step (3) The quantity of young leaves is 2-3 pieces.
- 7. the method for transformation of soybean adventitious bud rooting according to claim 1, it is characterised in that described in step (3) Continue also to include watering step during culture.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102304545A (en) * | 2011-09-07 | 2012-01-04 | 河北科技师范学院 | Method for converting soybeans by using agrobacterium |
| CN103329799A (en) * | 2013-06-08 | 2013-10-02 | 上海辰山植物园 | Rapid propagation and seedling growing method for Shuhua camellia tissue culture |
-
2015
- 2015-09-30 CN CN201510638870.6A patent/CN105230485B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102304545A (en) * | 2011-09-07 | 2012-01-04 | 河北科技师范学院 | Method for converting soybeans by using agrobacterium |
| CN103329799A (en) * | 2013-06-08 | 2013-10-02 | 上海辰山植物园 | Rapid propagation and seedling growing method for Shuhua camellia tissue culture |
Non-Patent Citations (1)
| Title |
|---|
| 影响大豆遗传转化再生植株成活因素的初步研究;李桂兰等;《河北农业大学学报》;20070331;第30卷(第2期);11-14 * |
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