CN109182375A - A kind of genetic transforming method of Iris germanica - Google Patents
A kind of genetic transforming method of Iris germanica Download PDFInfo
- Publication number
- CN109182375A CN109182375A CN201811199826.XA CN201811199826A CN109182375A CN 109182375 A CN109182375 A CN 109182375A CN 201811199826 A CN201811199826 A CN 201811199826A CN 109182375 A CN109182375 A CN 109182375A
- Authority
- CN
- China
- Prior art keywords
- iris germanica
- callus
- iris
- germanica
- transforming method
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
- C12N15/8205—Agrobacterium mediated transformation
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- General Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The method of the present invention discloses a kind of Iris germanica genetic transforming method of mediated by agriculture bacillus, belongs to Genetic Transformation in Higher Plants technical field.This method infects conversion mainly using the stem apex callus of Iris germanica as transformation receptor, by Agrobacterium, resistant calli screens, break up again and the processes such as the PCR of resistant plant detection obtain transgenic line.Method provided by the invention has easy to operate, the high advantage of genetic transformation rate, has great theory significance and application and popularization value for the initiative of the genetic transformation of Iris germanica or even other iris, gene functional research and new germ plasm.
Description
Technical field
The invention belongs to the constructing technology fields of Genetic Transformation in Higher Plants system, and in particular to a kind of heredity of Iris germanica turn
Change method.
Background technique
Iris germanica (Iris germanica L.) be Iridaceae Jris perennial herb, famous ornamental flower,
Have outside the changeable whisker of color on petal middle rib except the series of products, also with spending, big, flower pattern is peculiar, the beautiful in colour, four seasons
The characteristics such as evergreen are had huge development space and prospect in application on afforestation, potting and ground.Both at home and abroad in German kite
More research has been carried out in tail crossbreeding, tissue-culturing rapid propagation, cutflower preservation etc., and at present about utilize genetic engineering
The research that technology carries out genetic improvement to Iris germanica is rarely reported.Further investigate the iris such as Iris germanica flower correlation
Shape and degeneration-resistant molecular mechanism excavate related gene, and then cultivate the Iris germanica with merit (flower development, degeneration-resistant etc.)
New varieties are of great significance.The genetic conversion system for establishing Iris germanica is to disclose Iris germanica flower development and degeneration-resistant molecule machine
The key means of reason, it will be greatly promoted the Innovative process of the iris new germ plasm such as Iris germanica, the present invention is first
A kind of construction method of the secondary Iris germanica callus genetic conversion system for disclosing mediated by agriculture bacillus.
Summary of the invention
For the deficiency of traditional breeding technology, the problem to be solved in the present invention is to provide one kind can be used for Iris germanica molecule
Iris germanica genetic transforming method in breeding process.
To achieve the purpose of the present invention, the technical solution adopted by the present invention is that:
The method that a kind of pair of Iris germanica Transformation of Callus obtains transgenic plant, including Iris germanica callus
Induction and Multiplying culture, the preculture of callus, the preparation of agrobacterium strains containing plant expression vector, Agrobacterium bacterium solution are living
Change and the preparation of Agrobacterium infected liquid, Agrobacterium are infected and co-cultured, Agrobacterium takes off bacterium and evoking adventive bud regeneration, the training of adventitious bud
Support and take root, the screening of antibiotic positive seedling, the transplanting of antibiotic positive seedling and PCR identification and etc.；Specifically:
1, the induction and proliferation of Iris germanica callus: Iris germanica stem apex part is taken, after being cleaned with cleaning solution, is moved to
Superclean bench disinfection.Then stem apex is cut into small pieces and is placed on callus inducing medium, under light illumination callus induction group
It knits, and carries out squamous subculture.The callus inducing medium are as follows: MS+1.5mg/L 2,4-D+0.2mg/L 6-BA+30g/
L sucrose+6g/L agar powder, pH 5.8.
2, the fritter that subculture 2-3 times callus is cut into 2mm size the preculture of callus: is placed in Multiplying culture
On base, dark culture is spare after 5-7 days；The proliferation culture medium formula are as follows: MS+1.0mg/L2,4-D+0.1mg/L 6-BA+
30g/L sucrose+6g/L agar powder, pH 5.8.
3, Agrobacterium bacterium liquid activation and the preparation of Agrobacterium infected liquid: by the Agrobacterium containing plasmid in addition rifampin
It crosses on YEB solid medium, picking single colonie after 28 DEG C of dark culture 3d is linked into the YEB fluid nutrient medium containing rifampin, black
28 ± 2 DEG C under dark condition, thalline were collected by centrifugation by shaken cultivation 16h, and thallus is resuspended with 1/2MS solution, adjusts to OD600Value is
0.3-0.5 adds acetosyringone (200mg/L), obtains Agrobacterium infected liquid；Agrobacterium strains for infecting are
EHA105, the agrobacterium strains PBI121 containing plant expression vector；
4, Agrobacterium infects callus: the Iris germanica callus after preculture is invaded in the Agrobacterium containing plasmid
It carries out drying after infecting 3min in dye liquor, and is placed on and co-cultures on base, co-cultured under dark condition；5, Agrobacterium takes off bacterium and resists
Property callus screening: by step 4 co-culture after Iris germanica callus sterile water wash after, go to containing G418
On the callus proliferation medium of (100mg/L) and Tim (150mg/L), at 25 ± 2 DEG C, under the conditions of the photoperiod is 14/10h
Resistant calli is screened in culture 3 weeks；
6, kanamycin-resistant callus tissue evoking adventive bud regenerates: the resistant calli being newly proliferated in step 5 is gone to containing G418
On the adventitious bud induction culture base of (150mg/L) and Tim (150mg/L), evoking adventive bud is generated, and condition of culture is the same as step 5 institute
It states, after culture 4 weeks, differentiates adventitious bud；The adventitious bud induction culture based formulas are as follows: MS+1.0mg/L 6-BA+0.2mg/
LNAA+150mg/L G418+150mg/L Ticarcillin/Clavulanate Acid+30g/L sucrose+6g/L agar powder, pH 5.8.
7, resistant buds are taken root: when Iris germanica adventitious bud it is long to 3-4cm when, budlet is cut and is gone to containing G418 (150mg/
L) and on the root media of Tim (150mg/L), condition of culture is taken root after 3 weeks with step 6, culture；The culture of rootage basigamy
Side are as follows: MS+0.2mg/L NAA+150mg/L G418+150mg/L Ticarcillin/Clavulanate Acid+30g/L sucrose+6g/L agar powder, pH 5.8.
8, the transplanting of resistance seedling and transgenosis PCR identification: well-grown resistance seedling is taken out from culture medium, moves into battalion
It supports in soil, is cultivated in the greenhouse.The young leaves that clip Iris germanica kanamycin-resistant callus tissue induces is extracted DNA, is carried using PBI121
Special primer carries out PCR detection on body, screens positive transgenic seedling.
Compared with the existing technology, the application achieve it is following the utility model has the advantages that
The present invention provides a kind of construction methods of genetic conversion system of ornamental flower Iris germanica.This method is with Germany
Iris stem apex callus is transformation receptor, carries out genetic transformation by mediated by agriculture bacillus.Stem apex callus selected by the present invention
Organizing inductivity is 82%, and differentiation and regeneration ability is strong, and it is good that Agrobacterium infects effect, genetic transformation efficiency 50%.By this hair
The Iris germanica genetic conversion system of bright foundation can be used for the genetic improvement of Iris germanica, the Innovative of new germ plasm, also be it
Genetic transformation, molecular improvement of his iris etc. provide theory and technology reference frame.
The present invention combines Genomic PCR to identify using two kinds of antibiotic (G418 and Tim) screenings, and positive turn has successfully been obtained
Gene plant, and positive rate is 50%.
Detailed description of the invention
Fig. 1: Iris germanica stem apex callus；
Fig. 2: the screening of Iris germanica resistant calli；
Fig. 3: the differentiation of Iris germanica resistant calli；
Fig. 4: resistant plant is taken root；
Fig. 5: the PCR identification of transgenic plant.
M:DNA Marker in Fig. 5,1-4: resistance strain；5: plasmid positive control；6 WT strain negative controls.
Specific embodiment
Below with reference to embodiment, the invention will be further elaborated, and embodiment below facilitates a better understanding of this hair
It is bright, but do not limit the present invention.
Embodiment 1
The present invention is a kind of method of Iris germanica genetic conversion system building, material therefor and reagent: Iris germanica stem
Point, Agrobacterium (EHA105), plasmid (PBI121)；Dehydrated alcohol, mercuric chloride, MS culture medium, 2,4- dichlorphenoxyacetic acid (2,4-
D), α-naphthylacetic acid (NAA), 6-benzyl aminopurine (6-BA), rifampin (Rif), acetosyringone (AS), Geneticin
(G418), Ticarcillin/Clavulanate Acid (Tim), sucrose and agar powder.Capital equipment: illumination box, superclean bench, centrifuge, PCR amplification
Instrument.
Its step are as follows:
1, the induction and proliferation of Iris germanica callus: 2-3 raw Iris germanica plant is taken back from being dug nursery
Laboratory, cuts off blade and rhizome, retains away from stem apex about 1cm up and down, impregnates 10min with 10% cleaning solution, then flowing water rushes
1h is washed, superclean bench is moved to, with 75% ethanol disinfection 45s, with aseptic water washing 2 times, then 0.1% mercuric chloride disinfection
10min, aseptic water washing 5 times.The explant that stem apex is cut into 2 × 2 × 2mm size is set after then being absorbed water with aseptic filter paper
In on callus inducing medium, the illumination cultivation at 25 ± 2 DEG C.The wherein callus inducing medium formula are as follows:
MS+2,4-D (1.5mg/L)+6-BA (0.2mg/L)+sucrose (30g/L)+agar powder (6g/L), pH 5.8；It is gone to after induction 30d
Multiplying culture is carried out on proliferated culture medium, subculture cycle is 30d (Fig. 1).
2, the fritter that subculture 2-3 times callus is cut into 2mm size the preculture of callus: is placed in Multiplying culture
On base, 5-7d is cultivated under 25 ± 2 DEG C, dark condition, it is spare；The wherein callus proliferation medium formula are as follows: MS+2,
4-D (1.0mg/L)+6-BA (0.1mg/L)+sucrose (30g/L)+agar powder (6.0g/L), pH is adjusted to 5.8；
3, Agrobacterium bacterium liquid activation and the preparation of Agrobacterium infected liquid: by the Agrobacterium containing plasmid in addition rifampin
It crosses on the YEB solid medium of (50mg/L), picking single colonie after 28 DEG C of dark culturing 3d is linked into containing rifampin (50mg/
L YEB fluid nutrient medium), 28 ± 2 DEG C, 200-220rpm shaken cultivation 16h under dark condition；Bacterium solution after culture is placed in
In the centrifuge tube of sterilizing, 6000rpm is centrifuged 5min and collects thallus, and thallus is resuspended with 1/2MS solution, adjusts to OD600Value is 0.3-
0.5, it adds acetosyringone (200mg/L), obtains Agrobacterium infected liquid；Agrobacterium strains for infecting are that EHA105 (contains
Plant expression vector PBI121).
4, Agrobacterium infects callus: choosing yellow-white relatively dry, hard-packed Iris germanica callus exists
3min is infected in Agrobacterium infected liquid, bacterium solution is sucked with aseptic filter paper and is placed in co-culture medium, 25 under dark condition
DEG C culture 3d；Wherein, above-mentioned co-culture medium formula are as follows: MS+2,4-D (1.5mg/L)+6-BA (0.2mg/L)+sucrose
(30g/L)+agar powder (6.0g/L), pH is adjusted to 5.8；
5, Agrobacterium takes off bacterium and resistant calli screening: the Iris germanica callus after co-culturing in step 4 is used
It, goes on callus proliferation medium sterile water wash for 5 times, then blotted with aseptic filter paper, at 25 ± 2 DEG C, the photoperiod 14/
It is cultivated 3 weeks under the conditions of 10h；Above-mentioned callus proliferation medium formula are as follows: MS+2,4-D (1.0mg/L)+6-BA (0.1mg/L)
+ sucrose (30g/L)+agar powder (6.0g/L), wherein addition antibiotic G418 (100mg/L) and Ticarcillin/Clavulanate Acid Tim (150mg/L),
PH is adjusted to 5.8；As shown in Fig. 2, on resistance screening culture medium growth a period of time and, kanamycin-resistant callus tissue can grow new callus
Tissue, rather than then gradually browning is dead for kanamycin-resistant callus tissue.
6, kanamycin-resistant callus tissue evoking adventive bud regenerates: the callus being newly proliferated in step 5 (white) being gone to adventitious bud and is lured
It leads on culture medium, condition of culture is same as above, and after culture 4 weeks, differentiates adventitious bud (Fig. 3)；The adventitious bud induction culture based formulas
Are as follows: MS+NAA (0.2mg/L)+6-BA (1.0mg/L)+sucrose (30g/L)+agar powder (6.0g/L), wherein adding antibiotic
G418 (150mg/L) and Tim (150mg/L), pH is adjusted to 5.8；
7, resistant buds are taken root: when Iris germanica adventitious bud it is long to 3-4cm when, budlet is cut and is gone on root media,
Condition of culture is same as above, and culture took root (Fig. 4) after 3 weeks；The prescription of rooting medium are as follows: MS+NAA (0.2mg/L)+sucrose
(30g/L)+agar powder (6.0g/L), wherein addition antibiotic G418 (150mg/L) and Tim (150mg/L), pH is adjusted to 5.8；
8, the transplanting of resistance seedling and transgenosis PCR identification: after resistance seedling root growth is good, it is not injured to root and is taken
Out, it washes away remaining culture medium and moves into soil.4 transgenic lines are taken at random, grow 1-2 piece young leaves to seedling, clip is new respectively
Leaf with CTAB method extract DNA, using PBI121 vector specific primer (GUS-F:5 '-GCGGTAACAAGAAAGGGATC-3 and
NOS-R:5 '-AATCATCGCAAGACCGGC-3 ') PCR amplification is carried out, screen the resistance seedling of the PCR amplification positive.Such as Fig. 5 institute
Show in the 4 resistance strains randomly selected, there are 2 plants of seedling all to detect the pcr amplification product of about 250bp size, meet pre-
Phase size thereby determines that foreign gene has been integrated into Iris germanica genome, establishes the genetic conversion system of Iris germanica.
The stem apex Transformation of Callus method of mediated by agriculture bacillus established by the present invention has easy to operate, rapidly and efficiently etc.
Feature.The present invention combines Genomic PCR to identify using two kinds of antibiotic (G418 and Tim) screenings, and the positive has successfully been obtained and turns base
Because of plant, and positive rate is 50%.In conclusion the present invention establish it is a kind of using Iris germanica stem apex as the operation of explant
Simply, the Iris germanica plant genetic transforming method of high efficient and reliable, in this way, can be efficiently by the matter containing foreign gene
Grain is transferred to Iris germanica, obtains positive transgenic Iris germanica plant.
Embodiment 2
Take well-grown stem apex, young tender ovary, the tender petal of children, the tip of a root as explant respectively, remaining processing step and
It is identical in embodiment 1, counted when 45d the pollution rate of explant, Callus induction rate, callus state and color, as a result
As shown in table 1.
Influence of the different explants of table 1 to Iris germanica callus induction
Above-mentioned data are analyzed it is found that the present invention is using Iris germanica stem apex as explant, there is callus induction
Rate is up to the features such as 82%, embryo is good, material is easy to get.
Embodiment 3
Using the processing method in embodiment 1, difference is the acetosyringone and OD of Agrobacterium infected liquid600Value,
Positive rate is counted, the results are shown in Table 2.
The different acetosyringone concentration of table 2 and OD600Influence of the value to changing effect
Above-mentioned data are analyzed it is found that in the present invention Agrobacterium infected liquid acetosyringone be 200mg/L and
OD600Value be 0.4 when, positive rate highest.
Finally, it is stated that embodiment described above is only used to illustrate the technical scheme of the present invention rather than to the invention patent
The limitation of range, although the present invention has been described in detail by examples detailed above, those skilled in the art should
Understand, can it be made several improvements and be deformed in the form and details, be limited without departing from claims of the present invention
Fixed range.The scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of genetic transforming method of Iris germanica, which is characterized in that steps are as follows:
(1) it takes the stem apex of Iris germanica to be seeded to callus inducing medium, evoked callus, and carries out squamous subculture；
(2) the Iris germanica callus for taking step (1) to obtain, is placed on proliferated culture medium after dark culture, spare；
(3) the Iris germanica callus that step (2) obtains is infected in the Agrobacterium infected liquid containing plasmid, then
It is placed in co-culture medium, is co-cultured；
(4) after the Iris germanica callus cleaning after co-culturing in step (3), antibiotic callus proliferation is gone to
On culture medium, resistant calli is screened；
(5) resistant calli being newly proliferated in step (4) is gone on adventitious bud induction culture base, evoking adventive bud generates；
(6) Iris germanica adventitious bud budlet is cut and is gone on root media, root induction；
(7) well-grown resistance seedling is taken out from culture medium, moves into Nutrition Soil, clip resistance seedling young leaves DNA utilizes carrier
Special primer carries out PCR amplification, screens the resistance seedling of the PCR amplification positive.
2. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that in step (1), the callus
Organize induced medium are as follows: MS+1.5mg/L 2,4-D+0.2mg/L 6-BA+30g/L sucrose+6g/L agar powder, pH 5.8.
3. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that in step (2), the proliferation
Culture medium prescription are as follows: MS+1.0mg/L 2,4-D+0.1mg/L 6-BA+30g/L sucrose+6g/L agar powder, pH 5.8.
4. the genetic transforming method of Iris germanica according to claim 1, it is characterised in that in step (2), by subculture 2-3
The fritter that secondary callus is cut into 2mm size is placed on proliferated culture medium.
5. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that in step (3), for infecting
Agrobacterium strains be EHA105, the agrobacterium strains PBI121 containing plant expression vector, Agrobacterium infected liquid is added to
The acetosyringone of 200mg/L, OD600For 0.3-0.5.
6. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that in step (4), for Germany
The Ticarcillin/Clavulanate Acid of antibiotic G418 and 150mg/L containing 100mg/L in the culture medium of iris kanamycin-resistant callus tissue screening.
7. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that described indefinite in step (5)
Bud inducement cultivation based formulas are as follows: MS+1.0mg/L 6-BA+0.2mg/L NAA+150mg/LG418+150mg/L Ticarcillin/Clavulanate Acid+30g/
L sucrose+6g/L agar powder, pH 5.8.
8. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that described to take root in step (6)
Culture medium prescription are as follows: MS+0.2mg/LNAA+150mg/L G418+150mg/L Ticarcillin/Clavulanate Acid+30g/L sucrose+6g/L agar powder, pH
It is 5.8.
9. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that in step (7), clip Germany
The young leaves that iris kanamycin-resistant callus tissue induces extracts DNA, carries out PCR detection using special primer on PBI121 carrier, screening is positive
Transgenic seedlings.
10. the genetic transforming method of Iris germanica according to claim 1, which is characterized in that in step (3), Agrobacterium
Infected liquid the preparation method comprises the following steps: by the Agrobacterium containing plasmid addition 50mg/L rifampin YEB solid medium on cross, 28
Picking single colonie after DEG C dark culturing 3d, is linked into the YEB fluid nutrient medium of the rifampin containing 50mg/L, 28 ± 2 under dark condition
DEG C, 200-220rpm shaken cultivation 16h；Bacterium solution after culture is placed in the centrifuge tube of sterilizing, 6000rpm is centrifuged 5min and collects
Thallus is resuspended with 1/2MS solution in thallus, and adjusting to OD600 value is 0.3-0.5, adds 200mg/L acetosyringone, obtains agriculture
Bacillus infected liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811199826.XA CN109182375B (en) | 2018-09-26 | 2018-09-26 | Genetic transformation method of German iris |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811199826.XA CN109182375B (en) | 2018-09-26 | 2018-09-26 | Genetic transformation method of German iris |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109182375A true CN109182375A (en) | 2019-01-11 |
| CN109182375B CN109182375B (en) | 2022-02-15 |
Family
ID=64945143
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811199826.XA Active CN109182375B (en) | 2018-09-26 | 2018-09-26 | Genetic transformation method of German iris |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109182375B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111165357A (en) * | 2020-03-02 | 2020-05-19 | 东北林业大学 | Method for inhibiting endophyte pollution in callus proliferation process of cordyceps sobolifera |
| CN113736819A (en) * | 2021-09-02 | 2021-12-03 | 浙江大学 | Method for constructing butterfly flower gene silencing system |
| CN116965325A (en) * | 2023-08-14 | 2023-10-31 | 江苏省中国科学院植物研究所 | Method for obtaining hybrid seeds and seed germination of iris Netherlands |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107494267A (en) * | 2017-09-27 | 2017-12-22 | 江苏农林职业技术学院 | A kind of cultural method of Iris germanica callus |
-
2018
- 2018-09-26 CN CN201811199826.XA patent/CN109182375B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107494267A (en) * | 2017-09-27 | 2017-12-22 | 江苏农林职业技术学院 | A kind of cultural method of Iris germanica callus |
Non-Patent Citations (2)
| Title |
|---|
| ZORAN JEKNIC等: "Genetic Transformation of Iris germanica Mediated by Agrobacterium tumefaciens", 《J. AMER. SOC. HORT. SCI.》 * |
| 黄苏珍等: "德国鸢尾的组织培养", 《江苏林业科技》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111165357A (en) * | 2020-03-02 | 2020-05-19 | 东北林业大学 | Method for inhibiting endophyte pollution in callus proliferation process of cordyceps sobolifera |
| CN111165357B (en) * | 2020-03-02 | 2022-03-22 | 东北林业大学 | Method for inhibiting endophyte pollution in callus proliferation process of cordyceps sobolifera |
| CN113736819A (en) * | 2021-09-02 | 2021-12-03 | 浙江大学 | Method for constructing butterfly flower gene silencing system |
| CN113736819B (en) * | 2021-09-02 | 2023-11-03 | 浙江大学 | Construction method of butterfly flower gene silencing system |
| CN116965325A (en) * | 2023-08-14 | 2023-10-31 | 江苏省中国科学院植物研究所 | Method for obtaining hybrid seeds and seed germination of iris Netherlands |
| CN116965325B (en) * | 2023-08-14 | 2024-05-03 | 江苏省中国科学院植物研究所 | Method for obtaining hybrid seeds and seed germination of iris Netherlands |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109182375B (en) | 2022-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101611697B (en) | Virus removal and rapid propagation technology of sweet potato variety 'Shangshu 19' | |
| CN103966258B (en) | A kind of agriculture bacillus mediated cabbage type rape genetic transforming method | |
| CN109182375A (en) | A kind of genetic transforming method of Iris germanica | |
| CN104004781A (en) | Preparation method of glyphosate resistant transgenic rice | |
| Tang et al. | Callus induction and plant regeneration from in vitro cultured leaves, petioles and scales of Lilium leucanthum (Baker) Baker | |
| CN104542276B (en) | A kind of method of red palm virus-free culture | |
| CN106258976B (en) | A kind of tissue culturing fast seedling-cultivating method of mustard type rape | |
| CN103340180B (en) | Method for identifying resistance function of aphid-resisting genes through transforming hairy roots of soybean | |
| CN107190019B (en) | A kind of sinocalamus latiflorus method for transformation of mediated by agriculture bacillus | |
| CN101946708B (en) | Genetic transformation method using sweet sorghum young ear or young ear induced callus as explant | |
| CN102907328B (en) | Method for utilizing microelements to accelerate tissue culture seedling of sugarcane | |
| CN108396035A (en) | A kind of agriculture bacillus mediated rice transformation method | |
| CN110305894A (en) | A kind of Chinese catalpa genetic transforming method rapidly and efficiently | |
| CN108184661A (en) | A kind of highly effective revulsion induction method of red palm polyploid | |
| CN102210266B (en) | Culture medium for culturing lilium pumilum tissues | |
| CN100334203C (en) | Establishent of acacia crassicarpa high efficiency transforming system induced by agricultural bacillus | |
| CN105838734A (en) | Method for establishing ziziphus jujuba genetic transformation system for receptor by leaves | |
| CN105886527A (en) | Agrobacterium tumefaciens mediated transformation system for efficiently obtaining transgenic plants of paspalum vaginatum and application thereof | |
| CN113755521A (en) | Construction method of agrobacterium-mediated strawberry sweet-charming genetic transformation system | |
| CN103053421B (en) | Chinese pistache rapid propagation method | |
| CN106342680A (en) | Method suitable for cultivation of various genotype wheat microspores | |
| CN106258952B (en) | The method formed using the tender lateral root induction Ammopiptanthus mongolicus callus of Ammopiptanthus mongolicus children | |
| CN106636196B (en) | A kind of peanut method of optimization, efficient mediated by agriculture bacillus | |
| CN108277233A (en) | The method for inducing and cultivating of Mature Embryos of Rice callus | |
| CN108546711A (en) | A kind of genetic transformation method for soybean |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |