CN111183899A - A method for rapidly inducing hairy roots of safflower - Google Patents
A method for rapidly inducing hairy roots of safflower Download PDFInfo
- Publication number
- CN111183899A CN111183899A CN202010057269.9A CN202010057269A CN111183899A CN 111183899 A CN111183899 A CN 111183899A CN 202010057269 A CN202010057269 A CN 202010057269A CN 111183899 A CN111183899 A CN 111183899A
- Authority
- CN
- China
- Prior art keywords
- safflower
- hairy roots
- days
- hairy
- infection solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 244000020518 Carthamus tinctorius Species 0.000 title claims abstract description 52
- 235000003255 Carthamus tinctorius Nutrition 0.000 title claims abstract description 51
- 238000000034 method Methods 0.000 title claims abstract description 23
- 230000001939 inductive effect Effects 0.000 title claims abstract description 15
- 208000015181 infectious disease Diseases 0.000 claims abstract description 23
- 239000007788 liquid Substances 0.000 claims abstract description 20
- 241000589156 Agrobacterium rhizogenes Species 0.000 claims abstract description 14
- 108700008625 Reporter Genes Proteins 0.000 claims abstract description 11
- 239000002609 medium Substances 0.000 claims abstract description 8
- 238000012546 transfer Methods 0.000 claims abstract description 7
- 238000012216 screening Methods 0.000 claims abstract description 5
- 239000012879 subculture medium Substances 0.000 claims abstract description 3
- 241000589158 Agrobacterium Species 0.000 claims description 10
- 230000006698 induction Effects 0.000 abstract description 18
- 238000012258 culturing Methods 0.000 abstract description 8
- 239000000463 material Substances 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 14
- 239000001963 growth medium Substances 0.000 description 10
- 229930186147 Cephalosporin Natural products 0.000 description 9
- 229940124587 cephalosporin Drugs 0.000 description 9
- 150000001780 cephalosporins Chemical class 0.000 description 9
- 239000007787 solid Substances 0.000 description 9
- 230000009466 transformation Effects 0.000 description 9
- 238000010186 staining Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 241000628997 Flos Species 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000003321 amplification Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000010008 shearing Methods 0.000 description 3
- 239000012192 staining solution Substances 0.000 description 3
- JXCKZXHCJOVIAV-UHFFFAOYSA-N 6-[(5-bromo-4-chloro-1h-indol-3-yl)oxy]-3,4,5-trihydroxyoxane-2-carboxylic acid;cyclohexanamine Chemical compound [NH3+]C1CCCCC1.O1C(C([O-])=O)C(O)C(O)C(O)C1OC1=CNC2=CC=C(Br)C(Cl)=C12 JXCKZXHCJOVIAV-UHFFFAOYSA-N 0.000 description 2
- 102000053187 Glucuronidase Human genes 0.000 description 2
- 108010060309 Glucuronidase Proteins 0.000 description 2
- -1 alkyl glycols Chemical class 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- 244000186140 Asperula odorata Species 0.000 description 1
- 241000208838 Asteraceae Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 235000008526 Galium odoratum Nutrition 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 235000019485 Safflower oil Nutrition 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 101150054900 gus gene Proteins 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 229930013686 lignan Natural products 0.000 description 1
- 235000009408 lignans Nutrition 0.000 description 1
- 150000005692 lignans Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001197 polyacetylene Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000003813 safflower oil Substances 0.000 description 1
- 235000005713 safflower oil Nutrition 0.000 description 1
- 229930004725 sesquiterpene Natural products 0.000 description 1
- 150000004354 sesquiterpene derivatives Chemical class 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种快速诱导红花毛状根的方法。是将3‑4周的无菌苗，分别用侵染液A和侵染液B侵染8‑15分钟，然后进行共培养24‑72小时后，转移到筛选培养进行培养，10天‑25天以后从外植体中剪下生长快速的毛状根，转移到继代培养基上，15天以后转移到液体培养基上进行扩大摇培，30‑45天以后得到大量毛状根；所述的侵染液A为野生型发根农杆菌K599，B为转入GUS报告基因的K599发根农杆菌。本发明具有对红花毛状根快速的诱导，适用于多种红花品种毛状根的诱导,具有取材方便、适用性强、诱导速度快的有益效果。The invention discloses a method for rapidly inducing hairy roots of safflower. It is to infect 3-4 weeks of sterile seedlings with Infection Solution A and Infection Solution B for 8-15 minutes, and then co-cultivate for 24-72 hours, then transfer to screening culture for culturing for 10-25 days. Cut off the fast-growing hairy roots from the explants after a few days, transfer them to a subculture medium, transfer them to a liquid medium after 15 days for expanded shaking culture, and obtain a large number of hairy roots after 30-45 days; Said infection solution A is wild-type A. rhizogenes K599, and B is K599 A. rhizogenes transformed into a GUS reporter gene. The invention has the advantages of rapid induction of safflower hairy roots, is suitable for the induction of hairy roots of various safflower varieties, and has the beneficial effects of convenient material selection, strong applicability and fast induction speed.
Description
Technical Field
The invention relates to a method for inducing safflower hairy roots, in particular to a method for rapidly inducing safflower hairy roots.
Background
Carthami flos is dry flower of Carthamus tinctorius L of Compositae, and is called grass Carthamus tinctorius L, Woodward Variegatus (L.) Druce, and radix seu folium Linderae Strychnifoliae. It is pungent and warm in property, enters heart and liver meridians, and is a good herb for activating blood and dredging meridians, removing blood stasis and relieving pain. The chemical components of safflower mainly comprise flavonoids, alkaloids, polyacetylenes, spermidine, lignans, sesquiterpenes, organic acids, sterols, alkyl glycols, polysaccharides and other components. The compounds in safflower have wide pharmacological activity, not only have certain effect on cardiovascular and cerebrovascular systems, nervous systems, immune systems and the like of human, but also have various physiological activities of anti-inflammation, analgesia, anti-tumor, antioxidation and the like. The safflower oil is edible oil which is recognized in the world and has the functions of eating, health care and beauty treatment.
Just as safflower has so many chemical components and functions of treatment and health care, it has been more and more widely used. Research shows that in the method for gene function verification of safflower, arabidopsis thaliana or callus induction regeneration is generally adopted, the growth cycle is long, and the transformation efficiency is low. The hairy root has the advantages of short growth cycle, convenient culture, easy transformation, high transformation rate and the like, and particularly, the successful induction and related application of safflower in the hairy root are not reported at home and abroad at present.
At present, only 1 safflower plant is accepted in the world in China, but the introduction and breeding work of safflower varieties in China is late, and the biotechnological application and development of safflower are slow, so that the induction and gene transformation efficiency of safflower hairy roots is very necessary.
Disclosure of Invention
The invention aims to provide a method for rapidly inducing safflower hairy roots. The method has the advantages of rapid induction of the safflower hairy roots, suitability for induction of the hairy roots of various safflower varieties, convenient material acquisition, strong applicability and rapid induction speed.
At present, no report for constructing a genetic system of the hairy roots of the safflower exists, and the invention fills the blank of research in the field. The method can be used for quickly obtaining the hairy root system, and provides good help for quickly developing gene function verification research on safflower.
The technical scheme of the invention is as follows: a method for inducing safflower hairy root rapidly, it is 3-4 weeks aseptic seedling, infect liquid A and infect liquid B to infect 8-15 minutes separately, then transfer to and screen and cultivate after co-culturing for 24-72 hours, cut the hairy root growing rapidly from the explant after 10 days-25 days, transfer to the subculture medium, transfer to the liquid culture medium after 15 days and enlarge and shake and cultivate, get a large amount of hairy roots after 30-45 days; the infection liquid A is wild agrobacterium rhizogenes K599, and the infection liquid B is K599 agrobacterium rhizogenes transferred with GUS reporter genes.
In the method for rapidly inducing hairy roots of safflower, the infection solution A is obtained by centrifuging 50ml OD value (600)0.4-1.2 agrobacterium K599 and resuspending 0.5-2.0 times volume of MS liquid.
In the method for rapidly inducing the hairy roots of the safflower, the infection liquid B is obtained by centrifuging 50ml of the Agrobacterium K599 with a GUS reporter gene with an OD value of 600 of 0.4-1.2 and then carrying out heavy suspension by using MS liquid with 0.5-2.0 times of the volume of the MS liquid.
Compared with the prior art, the invention has the following beneficial effects:
1. the transformation efficiency can be identified by transferring GUS genes in hairy roots and carrying out GUS staining on the genes, and the method has very important significance for utilizing the hairy roots of the safflower. The safflower explant is induced by agrobacterium rhizogenes, wherein the agrobacterium rhizogenes strain is a key substance. K599 is one of agrobacterium rhizogenes, and the plant rhizogenesis can be induced by the literature report. We studied the induction time, concentration, culture time and the like of K599 Agrobacterium rhizogenes on safflower, constructed transformed K599 Agrobacterium rhizogenes with GUS reporter genes, and performed GUS staining on transformed hairy roots. Experimental research shows that wild type and transformed K599 agrobacterium rhizogenes can efficiently induce hairy roots to safflower when OD value is within 0.4-1.2, and the safflower with GUS gene can be dyed into blue.
2. Starting with the approaches of K599 agrobacterium rhizogenes, biology of transformation efficiency and the like, the invention deeply researches the induction conditions and transformation efficiency of the hairy roots and finds that the strain selection and OD value thereof, the co-culture time and the induction part are key factors in the induction process of the hairy roots of the safflower. The method is applied to the induction of safflower hairy roots, constructs transformed K599 agrobacterium rhizogenes with GUS reporter genes, and performs GUS staining on the transformed hairy roots and is used for researching the transformation efficiency. The invention uses chemical staining method to stain GUS, so that the transformation efficiency can be simply, conveniently and intuitively explored. And the method has the advantages of simple operation, high analysis speed and low price of used reagents.
The principle of "dyeing red flower hairy roots infected with B-infection solution and cultured with GUS" in examples 1-3 to stain with color and find blue "is that GUS (β -glucuronidase, β -D-glucuronidase) gene is a reporter gene commonly used at present, and its expression product β -Glucuronidase (GUS) is a hydrolase which catalyzes the hydrolysis of many β -glucoside ester substances, and it can decompose 5-bromo-4-chloro-3-indole- β -glucoside acid ester (X-gluc. htm' target ═ blank > X-gluc) into blue substances, and its detection method is simple, rapid, sensitive, stable, and has low background activity.
Experiments prove that:
in examples 1 to 3 of the present invention, 40 explants were infected, and the number of hair roots and induction rate of hypocotyls and cotyledons were counted as shown in table 1:
table 1 examples 1-3 table of explant infections
In conclusion, the method has the advantages of rapid induction of the safflower hairy roots, suitability for induction of the hairy roots of various safflower varieties, convenience in material acquisition, strong applicability and rapid induction speed.
Detailed Description
The present invention is further illustrated by the following examples, which are not to be construed as limiting the invention.
Example 1:
a method for rapidly inducing safflower hairy roots, comprising the steps of:
(1) and (3) immersing a dye solution A: 50ml OD value (600) 0.4-0.6 Agrobacterium K599 centrifugal, use 2.0 times volume MS liquid heavy suspension to get and mix for subsequent use.
(2) And (3) immersing a dye solution B: 50ml OD value (600) 0.4-0.6 Agrobacterium K599 with GUS reporter gene for centrifugation, using 2.0 times volume MS liquid to carry out heavy suspension, and mixing evenly for standby.
(3) Taking two bottles of 15-20-day-old safflower aseptic seedlings.
(4) One group was added to 50ml of the infection solution A, and both hypocotyls and cotyledons were completely immersed in the liquid for 10 minutes of infection. The other set was added to 50ml of the staining solution B.
(5) Adding into MS culture medium at 25 deg.C, and dark culturing for about 24 h.
(6) After the dark culture is finished, transferring MS solid medium containing 200mg/ml of cephalosporin for 10-25 days, and then differentiating hypocotyls and cotyledons to form hairy roots.
(7) Shearing off 2-4CM hairy root, adding 100mg/ml solid medium of cephalosporin MS, and screening and culturing. After 30 days, the hairy roots of the safflower with high growth speed are screened out and subcultured on an MS solid culture medium containing 50mg/ml of cephalosporin. Subculturing for 2-3 times to obtain detoxified hairy root of Carthami flos.
(8) Cutting off 2-4CM hairy root, and adding MS liquid culture medium for amplification culture. Thereby obtaining the safflower hairy roots which grow fast and have more lateral root branches.
(9) GUS staining of the hairy roots of the safflower infected and cultured by the B infection solution can dye colors, and the color turns blue.
Example 2:
a method for rapidly inducing safflower hairy roots, comprising the steps of:
(1) and (3) immersing a dye solution A: centrifugation was performed at 50ml OD (600)0.8 Agrobacterium K599 and resuspension was performed with an equal volume of MS fluid to obtain a uniform mixture.
(2) And (3) immersing a dye solution B: 50ml OD value (600)0.8 Agrobacterium K599 with GUS reporter gene for centrifugation and equal volume MS liquid for heavy suspension to obtain uniform mixture.
(3) Taking two bottles of safflower aseptic seedlings with age of about 25-30 days.
(4) One group was added to 50ml of the infection solution A, and both hypocotyls and cotyledons were completely immersed in the liquid for 15 minutes of infection. The other set was added to 50ml of the staining solution B.
(5) Adding into MS culture medium at 25 deg.C, and dark culturing for about 48 h.
(6) After the dark culture is finished, transferring MS solid medium containing 200mg/ml of cephalosporin for 10-25 days, and then differentiating hypocotyls and cotyledons to form hairy roots.
(7) Shearing off 2-4CM hairy root, adding 100mg/ml solid medium of cephalosporin MS, and screening and culturing. After two weeks, the hairy roots of the safflower with high growth speed are screened out and subcultured on an MS solid culture medium containing 50mg/ml of cephalosporin. Subculturing for 2-3 times to obtain detoxified hairy root of Carthami flos.
(8) Cutting off 2-4CM hairy root, and adding MS liquid culture medium for amplification culture. Thereby obtaining the safflower hairy roots which grow fast and have more lateral root branches.
(9) GUS staining of NILA infected and cultured with the B-infection solution enables staining and bluing to be found.
Example 3:
a method for rapidly inducing safflower hairy roots, comprising the steps of:
(1) and (3) immersing a dye solution A: centrifugation was performed by 50ml OD (600)1.0 Agrobacterium K599, and resuspension was performed by 0.5 times MS liquid to obtain a uniform mixture.
(2) And (3) immersing a dye solution B: 50ml OD (600) about 1.0 Agrobacterium K599 with GUS reporter gene for centrifugation and 0.5 volume MS liquid heavy suspension to mix.
(3) Taking two bottles of 35-42-day-old safflower aseptic seedlings.
(4) One group was added to 50ml of the infection solution A, and both hypocotyls and cotyledons were completely immersed in the liquid for 15 minutes of infection. The other set was added to 50ml of the staining solution B.
(5) Adding into MS culture medium at 25 deg.C, and dark culturing for about 72 h.
(6) After the dark culture is finished, transferring MS solid medium containing 200mg/ml of cephalosporin for 10-25 days, and then differentiating hypocotyls and cotyledons to form hairy roots.
(7) Shearing off 2-4CM hairy root, adding 100mg/ml solid medium of cephalosporin MS, and screening and culturing. After two weeks, the hairy roots of the safflower with high growth speed are screened out and subcultured on an MS solid culture medium containing 50mg/ml of cephalosporin. Subculturing for 2-3 times to obtain detoxified hairy root of Carthami flos.
(8) Cutting off 2-4CM hairy root, and adding MS liquid culture medium for amplification culture. Thereby obtaining the safflower hairy roots which grow fast and have more lateral root branches.
(9) GUS staining of the hairy roots of the safflower infected and cultured by the B infection solution can dye colors, and the color turns blue.
Claims (3)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010057269.9A CN111183899A (en) | 2020-01-19 | 2020-01-19 | A method for rapidly inducing hairy roots of safflower |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010057269.9A CN111183899A (en) | 2020-01-19 | 2020-01-19 | A method for rapidly inducing hairy roots of safflower |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111183899A true CN111183899A (en) | 2020-05-22 |
Family
ID=70684672
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010057269.9A Pending CN111183899A (en) | 2020-01-19 | 2020-01-19 | A method for rapidly inducing hairy roots of safflower |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111183899A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111937749A (en) * | 2020-08-25 | 2020-11-17 | 贵州大学 | Efficient induction and cultivation method of hairy roots of Uncaria |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009126896A2 (en) * | 2008-04-10 | 2009-10-15 | Monsanto Technology Llc | Methods and compositions for root knot nematode control |
| CN105087635A (en) * | 2015-08-14 | 2015-11-25 | 贵州大学 | Method for inducing hairy roots of anisodus acutangulus C.Y.Wu et C.Chen |
| CN106432497A (en) * | 2016-11-05 | 2017-02-22 | 吉林农业大学 | Method of using safflower carthamus suspension cells for producing CD20 antibody |
| CN107760712A (en) * | 2017-12-14 | 2018-03-06 | 湖南科技大学 | A kind of method of the rapid induction hairy root in rape and identification transformation efficiency |
| CN109576302A (en) * | 2018-11-14 | 2019-04-05 | 甘肃农业大学 | A kind of method of hairy of rapid induction Ethiopia rape |
| CN109735562A (en) * | 2019-02-22 | 2019-05-10 | 北京林业大学 | A method for constructing an effective root transgenic system for economical plants |
| CN111088262A (en) * | 2020-01-19 | 2020-05-01 | 贵州大学 | Method for improving flavone content of safflower hairy roots by transferring CTMYB1 gene |
-
2020
- 2020-01-19 CN CN202010057269.9A patent/CN111183899A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009126896A2 (en) * | 2008-04-10 | 2009-10-15 | Monsanto Technology Llc | Methods and compositions for root knot nematode control |
| CN105087635A (en) * | 2015-08-14 | 2015-11-25 | 贵州大学 | Method for inducing hairy roots of anisodus acutangulus C.Y.Wu et C.Chen |
| CN106432497A (en) * | 2016-11-05 | 2017-02-22 | 吉林农业大学 | Method of using safflower carthamus suspension cells for producing CD20 antibody |
| CN107760712A (en) * | 2017-12-14 | 2018-03-06 | 湖南科技大学 | A kind of method of the rapid induction hairy root in rape and identification transformation efficiency |
| CN109576302A (en) * | 2018-11-14 | 2019-04-05 | 甘肃农业大学 | A kind of method of hairy of rapid induction Ethiopia rape |
| CN109735562A (en) * | 2019-02-22 | 2019-05-10 | 北京林业大学 | A method for constructing an effective root transgenic system for economical plants |
| CN111088262A (en) * | 2020-01-19 | 2020-05-01 | 贵州大学 | Method for improving flavone content of safflower hairy roots by transferring CTMYB1 gene |
Non-Patent Citations (3)
| Title |
|---|
| JOHN R. PORTER, PH.D等: "Host Range and Implications of Plant Infection by Agrobacterium rhizogenes", 《CRITICAL REVIEWS IN PLANT SCIENCES》 * |
| 向太和等: "发根农杆菌K599对菊花活体转化及其高效再生", 《园艺学报》 * |
| 周立刚等: "植物毛状根的培养及其化学进展――1.植物毛状根的诱导形成与培养", 《天然产物研究与开发》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111937749A (en) * | 2020-08-25 | 2020-11-17 | 贵州大学 | Efficient induction and cultivation method of hairy roots of Uncaria |
| CN111937749B (en) * | 2020-08-25 | 2021-11-19 | 贵州大学 | High-efficiency induction and culture method of uncaria hairy roots |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105238693B (en) | A kind of conventional cryopreservation method of straw mushroom strain | |
| CN101352144B (en) | A kind of Ganoderma lucidum hybrid breeding method | |
| CN103088059A (en) | Efficient genetic transformation method of hybridized tulip tree | |
| CN111183899A (en) | A method for rapidly inducing hairy roots of safflower | |
| CN114410550B (en) | A microbial functional flora for increasing flavonoid glycoside content in Ginkgo biloba and its preparation and application | |
| CN108823143A (en) | A kind of cultural method of purple perilla high yield Rosmarinic acid suspension cell | |
| CN110172474B (en) | Induction and rapid propagation method for hairy roots of purple ginseng | |
| CN108707624A (en) | A kind of method and application using agrobacterium rhizogenes Fiber differentiation macleaya cordata hairy root | |
| CN103088058B (en) | Genetic transformation method of Chinese fir | |
| CN110499278A (en) | A method for isolation, purification and transient transformation of ginkgo leaf protoplasts | |
| CN101775374B (en) | Production method of porcine epidemic diarrhea virus | |
| CN107475287B (en) | A kind of eggplant genetic transformation method | |
| Zeynali et al. | Effect of elicitation on antioxidant activity and production of tropane alkaloids in Hyoscyamus reticulatus hairy root cultures | |
| CN117247891B (en) | Callus culture of Panax notoginseng and its hairy root induction method | |
| CN1149918C (en) | Method for transferring agrobacterium mediated plant germination seed gene | |
| Ibrahim et al. | Stimulation of oleandrin production by combined Agrobacterium tumefaciens mediated transformation and fungal elicitation in Nerium oleander cell cultures | |
| GB2632530A (en) | Method for gene expression by transient transformation of wild rice seed using Agrobacterium | |
| CN109576302A (en) | A kind of method of hairy of rapid induction Ethiopia rape | |
| CN109762838A (en) | A genetic transformation system of spinach hairy roots mediated by Agrobacterium rhizogenes | |
| CN103146638A (en) | Induced proliferation method for glehnia littoralis hairy roots | |
| CN101525597B (en) | New hepatitis A inactivated vaccine virus strain and method for culturing same | |
| CN105861546B (en) | A kind of gene transformation method that the grape summer is black | |
| CN113897383B (en) | Induction and culture method for hairy root system of ammopiptanthus mongolicus for improving formononetin yield | |
| CN119082200B (en) | An efficient Agrobacterium-mediated genetic transformation method for ophiopogon aviculare | |
| CN111088262A (en) | Method for improving flavone content of safflower hairy roots by transferring CTMYB1 gene |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200522 |