CN101960991A - Onion callus induction method and special culture medium thereof - Google Patents
Onion callus induction method and special culture medium thereof Download PDFInfo
- Publication number
- CN101960991A CN101960991A CN 201010545355 CN201010545355A CN101960991A CN 101960991 A CN101960991 A CN 101960991A CN 201010545355 CN201010545355 CN 201010545355 CN 201010545355 A CN201010545355 A CN 201010545355A CN 101960991 A CN101960991 A CN 101960991A
- Authority
- CN
- China
- Prior art keywords
- onion
- explant
- medium
- callus induction
- concentration
- 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
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to the field of plant tissue culture, and discloses an onion callus induction method and a special culture medium thereof. The onion callus induction method comprises the following step of: carrying out callus induction culture on an onion explant by using a B5 culture medium added with 2,4-D and 6-BA, wherein the sucrose concentration of B5 culture medium is 30g/L and the agar concentration of B5 culture medium is 8g/L, the pH value of culture medium is 5.8, the added 2,4-D concentration is 0.5-2.0mg/L, and the 6-BA concentration is 1.0-4.0mg/L. The selected onion explant has the advantages of simple and convenient disinfection and sterilization method, and low pollution rate after inoculation and high callus induction rate.
Description
Technical field
The invention belongs to the Plant Tissue Breeding field, relate to a kind of onion callus induction method and special culture media thereof.
Background technology
Onion belongs to Liliaceae (Liliaceae) allium, is one of important vegetables of common cultivated in the world wide, and it constitutes product organ with meat scale and bulbil, and nutrient content is abundant, is important health-care nutritive vegetables.In recent years, China's onion area under cultivation enlarges rapidly, and onion and fabricated product thereof have been exported to the many countries in America and Europe, Japan and Southeast Asia.But China onion produces the kind that goes up use and mostly is external kind greatly at present, and domestic onion is of less types, and the reason that causes this situation is that onion is the biennial herb plant, the floral organ structure is little, easily degenerate after the selfing, exist breeding cycle long in the breeding, problem such as the breeding rate is low.Yet utilize tissue culture technique, can improve breeding efficiency.
The used starting material kind of onion tissue culture is a lot, and stem apex, plateau, blade, scale, floral organ official rank are drawn materials.Fridberg once directly lured the different bud that becomes with the MS medium of additional NAA and KT from the onion scale.Jiang Luyan etc. (2003) as explant, through callus and somatic embryo approach, have obtained the regeneration plant of upper frequency with the onion bud, and the highest inductivity of its callus is 68.71%.
Since nineteen eighty-three, first genetically modified plants were come out, the development of plant gene engineering technology was advanced by leaps and bounds.Crop transgenosis aspects such as tobacco, tomato, capsicum have a large amount of bibliographical informations, and some genetically modified crops are used on producing.The existing report of onion transformation system, Eady etc. (1998) are explant with the immature embryo, have set up the regenerating system of high frequency, the highest induction frequency is 60%, and to have set up with the Agrobacterium tumefaciems be mediation, is the transformation system of marker gene with gfp and npt II, obtained transgenic regenerated plant.But do not see practical application at present yet.
Summary of the invention
The objective of the invention is to overcome the above-mentioned deficiency of prior art, a kind of onion callus induction method is provided, another object of the present invention provides this method special culture media.
Purpose of the present invention can be achieved through the following technical solutions:
A kind of onion callus induction method comprises and chooses the onion explant, adopts and adds 2, and the B5 medium of 4-D, 6-BA carries out the inducing culture of callus; Sucrose concentration is 30g/L in the described B5 medium, and agar concentration is 8g/L, and the pH value is 5.8,2 of interpolation, and 4-D concentration is 0.5～2.0mg/L, 6-BA concentration is 1.0～4.0mg/L.
Wherein, described explant is selected from onion immature inflorescence or onion involucre by the young flower bud in the uncracked inflorescence.
The immature inflorescence of the about 0.8-1.2cm of the preferred onion diameter of described explant.
The preferred onion involucre of described explant is the young flower bud in the inflorescence of 1.5-2.0cm by uncracked diameter.
Selecting described onion involucre is young flower bud in the inflorescence of 1.5-2.0cm during as explant by uncracked diameter, the consisting of of medium: described B5 medium+1.0mg/L 2,4-D+1.0mg/L 6-BA.
This abductive approach also comprises the cultivation pre-treatment: earlier with the inflorescence sterilization, tear the involucre quilt then off, get explant, sterilization respectively.
Described method for disinfection and sterilization is at first washing 0.5h with running water, and 70% alcohol disinfecting 1min is used in the flowing water flushing again, with 0.1% the mercuric chloride 10min that sterilizes, uses rinsed with sterile water at last 3～5 times then.
Described condition of culture is that temperature is 25 ± 1 ℃, and light application time is 16h.d
-1, intensity of illumination is 50 μ molm
-2S
-1
With the sucrose concentration is 30g/L, and agar concentration is 8g/L, and the pH value is that 5.8 B5 medium is a basal medium, adds 0.5～2.0mg/L 2,4-D, 1.0～4.0mg/L 6-BA.
It is 30g/L that described medium consists of with the sucrose concentration, and agar concentration is 8g/L, and the pH value is that 5.8 B5 medium is a basal medium, adds 1.0mg/L 2,4-D and 1.0mg/L 6-BA.
Beneficial effect:
The present invention utilizes the onion bud of different growing stage to be explant, the factor that influences the callus generation is studied, thereby set up a kind of onion callus induction method, utilized this method to induce the onion callus, it is 100% that the callus of induce rate of explant reaches as high as.The present invention provides a kind of explant and medium that is exclusively used in the onion callus induction by selection, the Optimum of culture medium of explant, and this combination can significantly improve the inductivity of callus.Abductive approach of the present invention has improved the inductivity of onion children flower bud callus, for the conversion and the polyploid in-vitro inducing of foreign gene are laid a good foundation.
It is simple and convenient that the selected explant of the present invention has method for disinfection and sterilization, and inoculation after stain rate is low; The advantage that callus induction rate is high.
Description of drawings
Fig. 1, explant A cultivate 8 all backs callus induction pictures.
Children flower bud picture when Fig. 2, explant B, C inoculation.
Callus induction picture when Fig. 3, explant B cultivated for 8 weeks, the upper right corner is an enlarged drawing.
Callus induction picture when Fig. 4, explant C cultivated for 8 weeks, the upper right corner is an enlarged drawing.
Embodiment
Embodiment 1
1.1 the acquisition of sterilizable material
Test used onion material and take from gardening institute of Agricultural University Of Nanjing experiment base, kind is " 9866 ".Adopt the immature inflorescence of the about 0.8-1.2cm of diameter, tear off involucre by after, together with rhachis it being cut into 6-8 fritter is explant, numbers A; The involucre that adopts the about 1.5-2.0cm of diameter is by uncracked inflorescence, and getting its young flower bud of leaving short handle (0.1-0.3cm) is explant, numbering B; The inflorescence of the about 3.0cm of diameter that the employing involucre has been ftractureed, getting its young flower bud of leaving short handle (0.1-0.3cm) is explant, numbering C.
With the inflorescence sterilization, tear the involucre quilt then off earlier, get explant A and B respectively.Get the young flower bud of leaving short handle, sterilization obtains explant C.Sterilization at first washes 0.5h with running water, floats decontamination, and the alcohol disinfecting 1min with 70% with 0.1% mercuric chloride sterilization 10min, uses rinsed with sterile water 3～5 times then again.The sterilizable material explant A, B, the C that obtain are inoculated on the medium.
1.2 medium
B5 is a minimal medium, and wherein sucrose concentration is 30g/L, and agar concentration is 8g/L, and the pH value is 5.8.2, the set concentration gradient of 4-D is 0.5,1.0,2.0mg/L, and the set concentration gradient of 6-BA is 1.0,2.0,4.0mg/L, adopts Orthogonal Experiment and Design.Explant A, B, C are inoculated into respectively in the glass culture dish (or vial) of 9cm, each ware is inoculated explant A4, each 20 of explant B and C, and children flower bud picture is seen Fig. 2 during explant B, C inoculation, 3 culture dishes of every processing, 3 repetitions.Inoculation is placed in the culturing room cultivates, every 4 all successive transfer culture once.
The experimental scheme of table 1 callus induction
1.3 condition of culture
It is 25 ± 1 ℃ that postvaccinal culture dish or vial are placed temperature, and light application time is 16h.d
-1, intensity of illumination is 50 μ molm
-2S
-1Condition under cultivate.
1.4 three kinds of explants were cultivated after 8 weeks, statistics is inoculated the callus quantity that the back generates respectively, and data are carried out statistical analysis.
1.4.1 the growing state of different explants on inducing culture
The process of explant A callus induction has tangible different with explant B and C.After the explant A inoculation, at first rhachis base portion and young flower bud grow up, and after 4 weeks of growth, young flower bud anthocaulus is expanded, and have the callus of little yellow or white to occur on rhachis and the young flower bud respectively.The young flower bud of explant B and C gives birth to slowly long after inoculation, and the part bud is open, has yellowish callus (seeing Fig. 1,3,4) to occur after around the growth.
1.4.2 different explants is to the influence of onion callus induction
Experimental result shows, different explants is because of the difference of its physiological status, providing under the situation of identical hormone concentration and media, and there is very big difference in the situation of its callus induction.Each explant of explant A all has the appearance of callus, and the callus of induce rate of explant is 100%, the quantity difference (Fig. 1) of the callus on each explant.The inducing of the callus of explant B and C sees Table 2 and table 3.By table 2,3 know, the average inductivity that explant B (involucre is by the young flower bud in the uncracked inflorescence) forms callus is 85.49%, and the average inductivity of explant C (the young flower bud in the inflorescence that involucre is ftractureed) is 55.78%.Under identical condition of culture, the callus of induce rate of the processing of explant B all is higher than the processing of explant C.Illustrate that explant A is the induced material that immature inflorescence and the explant B onion children flower bud material that to be involucre got by uncracked inflorescence should be best callus.
The callus of induce situation of table 2 explant B
The callus of induce situation of table 3 explant C
1.4.3 different hormone concentration and media are to the influence of callus of induce situation
Explant is under different hormone kinds and concentration proportioning, and the situation of inducing of its callus has very big difference.Experimental result shows that explant A is in the processing of each, and inductivity is 100%, so explant A more easily induces callus, hormone concentration and media is little to its influence.In the processing of explant B, by interpretation is shown, 2, when 4-D was 0.5mg/L, 1.0mg/L, 2.0mg/L respectively, the average inductivity of callus was respectively 82.74%, 90.21%, 83.54%.When 6-BA was 1.0mg/L, 2.0mg/L, 4.0mg/L respectively, the average inductivity of callus was respectively 89.29%, 81.37%, 85.81%.Hence one can see that, and 2,4-D is 1.0mg/L, and 6-BA is that 1.0mg/L should be the optium concentration of explant B callus induction.On B5 medium, add 1.0mg/L 2,4-D+1.0mg/L in the processing of 6-BA, the inductivity of explant B callus is 98.33%, being significantly higher than the inductivity when only considering single hormone, is to be to be that the best hormone of explant induction onion callus is selected and concentration proportioning with involucre by the young flower bud in the uncracked inflorescence.In the processing of explant C, interpretation is shown when 2, when 4-D was 0.5mg/L, 1.0mg/L, 2.0mg/L respectively, the average inductivity of callus was respectively 46.99%, 55.90%, 64.44%.When 6-BA was 1.0mg/L, 2.0mg/L, 4.0mg/L respectively, the average inductivity of callus was respectively 43.56%, 61.01%, 62.77%.Hence one can see that, and 2,4-D is 2.0mg/L, 6-BA is that the best that 4.0mg/L should be explant C callus is induced hormone concentration.What yet explant C callus induction rate was the highest is to add 2, the processing of 4-D 1.0mg/L+6-BA4.0mg/L, and the healing rate of callus is 75.71%.The inducing of this phenomenon of inductivity when the inductivity of callus under single hormone condition is lower than two plant growth hormone comprehensive functions by explant B and C explanation callus is not to be determined by a certain hormone concentration, but the result of multiple hormone comprehensive function.
All are more satisfactory explant materials from young as can be seen tender inflorescence of above-mentioned experimental result and involucre by the young flower bud the uncracked inflorescence, when being explant by the young flower bud in the uncracked inflorescence with involucre, the best group of medium becomes: described B5 medium+1.0mg/L 2,4-D+1.0mg/L 6-BA.
Embodiment 2
Culture medium prescription: with the sucrose concentration is 30g/L, and agar concentration is 8g/L, and the pH value is that 5.8 B5 is a minimal medium, adds 1.0mg/L 2,4-D and 1.0mg/L 6-BA.
Get the immature inflorescence of the about 1cm of onion diameter, (at first wash 0.5h with running water, float decontamination, the alcohol disinfecting 1min with 70% with 0.1% mercuric chloride sterilization 10min, uses rinsed with sterile water 3～5 times to sterilization then again.), tear off involucre by after, together with rhachis it being cut into 6-8 fritter is explant A, with the acquisition sterilizable material explant A be inoculated on the medium.
It is 25 ± 1 ℃ that postvaccinal culture dish is placed temperature, and light application time is 16h.d
-1, intensity of illumination is 50 μ molm
-2S
-1Condition under cultivate.
After can observing inoculation, at first rhachis base portion and young flower bud grow up, and after 4 weeks of growth, young flower bud anthocaulus is expanded, and have the callus of little yellow and white to occur on rhachis and the young flower bud respectively.Each explant of cultivating 8 week back explant A all has the appearance of callus, and the callus of induce rate of explant is 100%, the quantity difference of the callus on each explant.
Embodiment 3
Culture medium prescription: with the sucrose concentration is 30g/L, and agar concentration is 8g/L, and the pH value is that 5.8 B5 is a minimal medium, adds 1.0mg/L 2,4-D and 1.0mg/L 6-BA.
Directly the involucre of about 1.5-2.0cm is by uncracked inflorescence for cut-off, and (at first wash 0.5h with running water, float decontamination, the alcohol disinfecting 1min with 70% with 0.1% mercuric chloride sterilization 10min, uses rinsed with sterile water 3～5 times to sterilization then again.), tear off involucre by after, getting its young flower bud of leaving short handle (0.1-0.3cm) is explant B, and the sterilizable material explant B that obtains is inoculated on the medium.
It is 25 ± 1 ℃ that postvaccinal culture dish is placed temperature, and light application time is 16h.d
-1, intensity of illumination is 50 μ molm
-2S
-1Condition under cultivate.
Can observe poor growth after inoculation, the part bud is open, has yellowish callus to occur after around the growth.After cultivating for 8 weeks, the callus of induce rate of explant B is 98.33%.
Claims (10)
1. an onion callus induction method is characterized in that comprising and chooses the onion explant, and adopt and add 2, the B5 medium of 4-D, 6-BA carries out the inducing culture of callus; Sucrose concentration is 30g/L in the described B5 medium, and agar concentration is 8g/L, and the medium pH value is 5.8,2 of interpolation, and 4-D concentration is 0.5～2.0mg/L, 6-BA concentration is 1.0～4.0mg/L.
2. onion callus induction method according to claim 1 is characterized in that described explant is selected from onion immature inflorescence or onion involucre by the young flower bud in the uncracked inflorescence.
3. onion callus induction method according to claim 2 is characterized in that described explant selects the immature inflorescence of onion diameter 0.8-1.2cm.
4. onion callus induction method according to claim 2 is characterized in that it is young flower bud in the inflorescence of 1.5-2.0cm by uncracked diameter that described explant is selected the onion involucre.
5. onion callus induction method according to claim 4, it is characterized in that selecting described onion involucre is that young flower bud in the inflorescence of 1.5-2.0cm is during as explant by uncracked diameter, consisting of of medium: described B5 medium+1.0mg/L 2,4-D+1.0mg/L 6-BA.
6. according to each described onion callus induction method in the claim 1～5, it is characterized in that this abductive approach also comprises the cultivation pre-treatment: earlier with the inflorescence sterilization, tear the involucre quilt then off, get explant, sterilization respectively.
7. onion callus induction method according to claim 6, it is characterized in that described method for disinfection and sterilization at first washing 0.5h with running water, 70% alcohol disinfecting 1min is used in the flowing water flushing again, with 0.1% mercuric chloride sterilization 10min, use rinsed with sterile water at last 3～5 times then.
8. according to each described onion callus induction method in the claim 1～5, it is characterized in that described condition of culture is that temperature is 25 ± 1 ℃, light application time is 16h/d, and intensity of illumination is 50 μ molm
-2S
-1
9. induce onion callus medium for one kind, it is characterized in that this medium consists of: be 30g/L with the sucrose concentration, agar concentration is 8g/L, the pH value is that 5.8 B5 medium is a basal medium, add 0.5～2.0mg/L 2,4-D, 1.0～4.0mg/L6-BA.
10. the onion callus medium of inducing according to claim 9, it is characterized in that it is 30g/L that described medium consists of with the sucrose concentration, agar concentration is 8g/L, and the pH value is that 5.8 B5 medium is a basal medium, add 1.0mg/L2,4-D and 1.0mg/L 6-BA.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010545355 CN101960991A (en) | 2010-11-15 | 2010-11-15 | Onion callus induction method and special culture medium thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010545355 CN101960991A (en) | 2010-11-15 | 2010-11-15 | Onion callus induction method and special culture medium thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101960991A true CN101960991A (en) | 2011-02-02 |
Family
ID=43514190
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010545355 Pending CN101960991A (en) | 2010-11-15 | 2010-11-15 | Onion callus induction method and special culture medium thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101960991A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102870674A (en) * | 2012-09-05 | 2013-01-16 | 广州白云华南生物科技有限公司 | Method for quickly propagating red onion through tissue culture |
| CN104488717A (en) * | 2014-12-22 | 2015-04-08 | 北京市农林科学院 | Method for obtaining multiple-ploidy regeneration plant by culturing flower bud of green Chinese onion |
| CN105941162A (en) * | 2016-06-27 | 2016-09-21 | 武汉生物工程学院 | Establishment method of high-efficiency regeneration system for allium cepa |
| CN107027633A (en) * | 2017-06-19 | 2017-08-11 | 东北林业大学 | A kind of purple skin onion adventitious root suspension culture method for improving prostaglandin A content |
| CN109156348A (en) * | 2018-09-16 | 2019-01-08 | 连云港市农业科学院 | A kind of method of onion sterile line callus induction |
| CN110226517A (en) * | 2019-06-26 | 2019-09-13 | 北京市农林科学院 | A kind of method of onion Regeneration in Vitro and its culture medium used |
| CN117441617A (en) * | 2023-12-22 | 2024-01-26 | 北京花乡花木集团有限公司 | Tissue culture method of north shallot |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1214857A (en) * | 1998-09-18 | 1999-04-28 | 北京市海淀区植物组织培养技术实验室 | Method for cultivating onion male sterile asexual line and conservation line |
-
2010
- 2010-11-15 CN CN 201010545355 patent/CN101960991A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1214857A (en) * | 1998-09-18 | 1999-04-28 | 北京市海淀区植物组织培养技术实验室 | Method for cultivating onion male sterile asexual line and conservation line |
Non-Patent Citations (3)
| Title |
|---|
| 《Plant Science》 19951231 Borut Bohanec等 Studies of gynogenesis in onion (Allium cepa L.): induction procedures and genetic analysis of regenerants 第215-224页 1-10 第104卷, 2 * |
| 《华北农学报》 20051231 杜敏霞，刘湘萍 洋葱幼蕾离体培养与植株再生的研究 第54-56页 1-10 第20卷, 第4期 2 * |
| 《山东农业大学学报（自然科学版）》 20031231 姜璐琰等 洋葱组织培养研究进展 第299-302页 1-10 第34卷, 第2期 2 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102870674A (en) * | 2012-09-05 | 2013-01-16 | 广州白云华南生物科技有限公司 | Method for quickly propagating red onion through tissue culture |
| CN102870674B (en) * | 2012-09-05 | 2013-12-25 | 广州白云华南生物科技有限公司 | Method for quickly propagating red onion through tissue culture |
| CN104488717A (en) * | 2014-12-22 | 2015-04-08 | 北京市农林科学院 | Method for obtaining multiple-ploidy regeneration plant by culturing flower bud of green Chinese onion |
| CN105941162A (en) * | 2016-06-27 | 2016-09-21 | 武汉生物工程学院 | Establishment method of high-efficiency regeneration system for allium cepa |
| CN105941162B (en) * | 2016-06-27 | 2018-04-24 | 武汉生物工程学院 | The method for building up of tillered-onion high-efficiency regeneration system |
| CN107027633A (en) * | 2017-06-19 | 2017-08-11 | 东北林业大学 | A kind of purple skin onion adventitious root suspension culture method for improving prostaglandin A content |
| CN109156348A (en) * | 2018-09-16 | 2019-01-08 | 连云港市农业科学院 | A kind of method of onion sterile line callus induction |
| CN110226517A (en) * | 2019-06-26 | 2019-09-13 | 北京市农林科学院 | A kind of method of onion Regeneration in Vitro and its culture medium used |
| CN110226517B (en) * | 2019-06-26 | 2021-06-01 | 北京市农林科学院 | In-vitro regeneration method of onion and culture medium used by same |
| CN117441617A (en) * | 2023-12-22 | 2024-01-26 | 北京花乡花木集团有限公司 | Tissue culture method of north shallot |
| CN117441617B (en) * | 2023-12-22 | 2024-04-02 | 北京花乡花木集团有限公司 | Tissue culture method of north shallot |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102893870B (en) | Rapid propagation and seedling raising method for beautiful millettia root seedling tissue culture | |
| CN101822220B (en) | Method for culturing and rapidly propagating stem tip tissue of rare cymbidium goeringii | |
| CN101960991A (en) | Onion callus induction method and special culture medium thereof | |
| CN102119655B (en) | Natural light rapid breeding method for dendrobium officinale | |
| CN102907327A (en) | Tissue culture propagation method for anoectochilus roxburghii | |
| CN103202229B (en) | Tissue culturing and rapid propagating method for chloranthy florida var. plena | |
| CN104082148A (en) | Method for performing regeneration propagation on sterile stalks of butterfly orchids | |
| CN101984796B (en) | Method for culturing large pearls by using clam visceral sac implanted nucleuses | |
| CN101720670A (en) | Rapid breeding method for pinellia tuber tissue culture | |
| CN103704130A (en) | Chinese orchid and cymbidium hybridum hybrid seedling raising method | |
| CN102613083A (en) | North American redwood tissue cultivation method | |
| CN102405841B (en) | Method for inducing callus of lycoris radiate by using flower stalks | |
| CN103651141B (en) | The method that Bo chrysanthemum batch production test tube seedling is the most numerous | |
| CN100394845C (en) | In-bottle production method of detoxified small seed ball of east lily | |
| CN106922536A (en) | A kind of method that fast energy-saving cultivates bletilla seedling | |
| CN104488722B (en) | A kind of quick breeding method for tissue culture of South America crutch flower | |
| CN104719168B (en) | Method for cultivating bletilla striata seedlings by using intermittent immersion bioreactor | |
| CN101766123B (en) | Method for rapid propagation of zephyr lily | |
| CN105766655A (en) | Establishing method of Gleditsia vestita Chun et Howex B.G.Li.tissue culture regeneration system | |
| CN105557515B (en) | A kind of tissue culture and rapid propagation method of roundleaf new pteris fern | |
| CN102232359B (en) | In-vitro rapid propagation method of double-petal Jasminum sambac | |
| CN106416972A (en) | Method for quickly cultivating bletilla striata seedlings | |
| CN106718875A (en) | Method for growing dendrobium seedlings | |
| CN104737913B (en) | The asexual rapid propagation method of Lepidinm meyenii Walp cenospecies | |
| CN106718878A (en) | A kind of fan fern quick breeding by group culture method with young sporangiorus as explant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20110202 |