CN116472961A - Application of squalene in Mo Langen-like stem subculture - Google Patents
Application of squalene in Mo Langen-like stem subculture Download PDFInfo
- Publication number
- CN116472961A CN116472961A CN202310448566.XA CN202310448566A CN116472961A CN 116472961 A CN116472961 A CN 116472961A CN 202310448566 A CN202310448566 A CN 202310448566A CN 116472961 A CN116472961 A CN 116472961A
- Authority
- CN
- China
- Prior art keywords
- squalene
- subculture
- langen
- stems
- culture
- 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
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 title claims abstract description 28
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 title claims abstract description 28
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 title claims abstract description 28
- 229940031439 squalene Drugs 0.000 title claims abstract description 28
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 title claims abstract description 28
- 239000001963 growth medium Substances 0.000 claims abstract description 27
- 230000035755 proliferation Effects 0.000 claims description 12
- 229920001817 Agar Polymers 0.000 claims description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 241000173252 Cymbidium sinense Species 0.000 claims description 8
- 238000005286 illumination Methods 0.000 claims description 8
- 241000272194 Ciconiiformes Species 0.000 claims description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- 239000012879 subculture medium Substances 0.000 claims description 6
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 claims description 2
- 239000005971 1-naphthylacetic acid Substances 0.000 claims description 2
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000000034 method Methods 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- JLIDBLDQVAYHNE-YKALOCIXSA-N (+)-Abscisic acid Chemical compound OC(=O)/C=C(/C)\C=C\[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-YKALOCIXSA-N 0.000 abstract description 4
- 150000003505 terpenes Chemical class 0.000 abstract description 4
- XHSDUVBUZOUAOQ-WJQMYRPNSA-N (3e,3ar,8bs)-3-[[(2r)-4-methyl-5-oxo-2h-furan-2-yl]oxymethylidene]-4,8b-dihydro-3ah-indeno[1,2-b]furan-2-one Chemical compound O1C(=O)C(C)=C[C@@H]1O\C=C/1C(=O)O[C@@H]2C3=CC=CC=C3C[C@@H]2\1 XHSDUVBUZOUAOQ-WJQMYRPNSA-N 0.000 abstract description 2
- 229930191978 Gibberellin Natural products 0.000 abstract description 2
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical group CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 abstract description 2
- 230000000975 bioactive effect Effects 0.000 abstract description 2
- 230000015572 biosynthetic process Effects 0.000 abstract description 2
- 150000001647 brassinosteroids Chemical class 0.000 abstract description 2
- FCRACOPGPMPSHN-UHFFFAOYSA-N desoxyabscisic acid Natural products OC(=O)C=C(C)C=CC1C(C)=CC(=O)CC1(C)C FCRACOPGPMPSHN-UHFFFAOYSA-N 0.000 abstract description 2
- 230000000694 effects Effects 0.000 abstract description 2
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 abstract description 2
- 239000003448 gibberellin Substances 0.000 abstract description 2
- 230000008121 plant development Effects 0.000 abstract description 2
- 230000008635 plant growth Effects 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- -1 brassinosteroids Chemical class 0.000 abstract 1
- 241000732800 Cymbidium Species 0.000 description 9
- 241000233855 Orchidaceae Species 0.000 description 8
- 239000000306 component Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 235000002779 Morchella esculenta Nutrition 0.000 description 1
- 240000002769 Morchella esculenta Species 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 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
- A01H4/002—Culture media 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
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses an application of squalene in Mo Langen like stems subculture. The secondary culture medium contains squalene. Squalene is an important bioactive substance with an all-trans isoprene structure, and participates in biosynthesis of terpenoids such as brassinosteroids, while terpenoids such as gibberellin, abscisic acid, strigolactone and the like are generally considered to have the effect of regulating plant growth and development.
Description
Technical Field
The invention relates to a plant tissue culture method in agricultural biotechnology, in particular to an application of squalene in Mo Langen like stem subculture.
Background
Orchid (Cymbidium) is a traditional Chinese famous flower and has important ornamental, cultural and economic values. The tissue culture and rapid propagation characteristics of different orchids are different, and the efficiency difference is obvious. The industrial production of the seedlings is one of the core technologies of the orchid industry and is one of the dominant seed industry fields in China, and the benefits of orchid breeding and seedling production are directly affected. Since Morel (1960) first uses the stem tip of cymbidium hybridum as an explant to induce protocorm-like bodies and differentiate the protocorm-like bodies into plants, a tissue culture rapid propagation system of cymbidium hybridum, spring sword, cymbidium hybridum and other national orchids is established successively. However, cymbidium represented by cymbidium is still one of the most difficult species of orchids to be industrially propagated at present, and one of the main reasons is that Mo Langen stems grow slowly and have low propagation coefficients. Therefore, the intensive research on the tissue culture and rapid propagation characteristics of orchids has very important practical significance for promoting the research and industrial development of orchids in China.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides application of squalene in Mo Langen-shaped stem subculture.
The invention is realized by the following technical scheme: use of squalene in Mo Langen corm subculture, and inoculating Mo Langen corms into squalene-containing culture medium for proliferation culture.
In one embodiment of the invention, the secondary culture medium contains 50-100 mg of squalene, 0.1-0.5 mg of 6-benzylaminopurine, 0-2.0 mg of 1-naphthylacetic acid, 20-30 g of sucrose, 5-7 g of agar powder, and the balance of MS culture medium, wherein the pH value is 5.4-6.0.
The proliferation culture conditions are as follows: inoculating Mo Langen stems into a secondary culture medium, performing dark culture at 25-28 ℃ for 10-15 days, transferring to the culture temperature of 25-28 ℃ and the illumination intensity of 1000-1500lx, and culturing for 25-30 days for 12 hours/day.
In the preparation of the secondary culture medium, squalene is dissolved in ethyl acetate, and is added after filtration sterilization and cooling of the secondary culture medium to below 50 ℃.
The Mo Langen corms are root corms of the cymbidium sinense variety 'penguin Bai Momo blue' with the length of 1.0-2.0 cm and the stem tip removed.
The invention applies squalene to the subculture of the cymbidium sinense, which can obviously promote the proliferation of the cymbidium sinense and accelerate the proliferation speed. Terpenoids such as gibberellin, abscisic acid, strigolactone, and the like are generally considered to have the effect of regulating plant growth and development, and Squalene (CAS number 111-02-4) is an important bioactive substance with an all-trans isoprene structure, and is involved in biosynthesis of terpenoids such as brassinosteroids. The invention adds squalene in the cymbidium subculture medium to obviously promote the proliferation of cymbidium, and the combined action of squalene and exogenous hormone promotes the proliferation of cymbidium, thereby improving the proliferation coefficient, shortening the subculture period, reducing the production cost and having good application prospect.
The invention has the advantages of unique use of the secondary culture medium, low cost, high propagation coefficient, short secondary period, good quality of the rhizomes and good application prospect.
Detailed Description
The technical scheme of the present invention will be described in further detail below with reference to the specific embodiments, but the present invention is not limited thereto.
Example 1
The cymbidium sinense variety 'penguin Bai Momo blue' root stems with the length of 1.0-2.0 cm and stem tip removed are respectively inoculated into the secondary culture mediums 1-3, firstly, dark culture is carried out for 10 days at the culture temperature of 25 ℃, then, the culture temperature is changed to 25 ℃, the illumination intensity is 1000lx, the illumination time is 12 hours/day, the culture is carried out for 25 days, and the proliferation coefficients of the 'penguin Bai Momo blue' root stems cultured in the secondary culture mediums 1-3 are 3.05, 0.47 and 2.18 respectively. Wherein the culture medium component of the secondary culture medium 1 is MS+50mg/L squalene+0.1 mg/L6-BA+1.0 mg/L NAA+20g/L sucrose+5 g/L agar, and the pH value is 5.4; the culture medium of the secondary culture medium 2 comprises MS+0.1 mg/L6-BA+1.0 mg/L NAA+0g/L sucrose+5 g/L agar, and the pH value is 5.4; the culture medium of the secondary culture medium 3 comprises MS+20mg/L squalene+0.1 mg/L6-BA+1.0 mg/L NAA+0g/L sucrose+5 g/L agar, and has the pH value of 5.4.
Example 2
The cymbidium sinense variety 'penguin Bai Momo blue' root-like stems with the length of 1.0-2.0 cm and stem tip removed are respectively inoculated into a secondary culture medium 4-5, firstly, dark culture is carried out for 13 days at the culture temperature of 27 ℃, then, the culture temperature is changed to 27 ℃, the illumination intensity is 1200lx, the illumination time is 12 hours/day, the culture is carried out for 27 days, and the proliferation coefficients of the cymbidium sinense Bai Momo blue root-like stems cultured in the secondary culture medium 4-5 are 3.39 and 0.56 respectively. Wherein the culture medium component of the secondary culture medium 4 is MS+80mg/L squalene+0.3 mg/L6-BA+1.5 mg/L NAA+25g/L sucrose+6 g/L agar, and the pH value is 5.7; the medium composition of the secondary culture medium 5 is MS+0.3 mg/L6-BA+1.5 mg/L NAA+25g/L sucrose+6 g/L agar, and the pH value is 5.7.
Example 3
The cymbidium sinense variety 'penguin Bai Momo blue' root stems with the length of 1.0-2.0 cm and stem tip removed are respectively inoculated into secondary culture mediums 6-8, firstly, dark culture is carried out for 15 days at the culture temperature of 28 ℃, then, the culture temperature is changed to 28 ℃, the illumination intensity is 1500lx, the illumination time is 12 hours/day, the culture is carried out for 30 days, and the proliferation coefficients of the 'penguin Bai Momo blue' root stems cultured in the secondary culture mediums 6-8 are 3.84, 0.61 and 2.69 respectively. Wherein the culture medium composition of the secondary culture medium 6 is MS+100mg/L squalene+0.5 mg/L6-BA+2.0 mg/L NAA+30g/L sucrose+7 g/L agar, and the pH value is 6.0; the culture medium composition of the secondary culture medium 7 is MS+0.5 mg/L6-BA+2.0 mg/L NAA+30g/L sucrose+7 g/L agar, and the pH value is 6.0; the culture medium of the secondary culture medium 8 comprises MS+120mg/L squalene+0.5 mg/L6-BA+2.0 mg/L NAA+30g/L sucrose+7 g/L agar, and has the pH value of 6.0.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (5)
1. The application of squalene in the subculture of Mo Langen corms, and the Mo Langen corms are inoculated in a subculture medium for proliferation culture, and the method is characterized in that the subculture medium contains squalene.
2. The use of squalene according to claim 1 in the subculture of Mo Langen corm, wherein the subculture medium comprises 50-100 mg of squalene, 0.1-0.5 mg of 6-benzylaminopurine, 0-2.0 mg of 1-naphthylacetic acid, 20-30 g of sucrose, 5-7 g of agar powder and the balance of MS culture medium, and the pH value is 5.4-6.0.
3. The use of squalene according to claim 1 in the subculture of Mo Langen like stems, wherein the proliferation culture conditions are: inoculating Mo Langen stems into a secondary culture medium, performing dark culture at 25-28 ℃ for 10-15 days, transferring to the culture temperature of 25-28 ℃ and the illumination intensity of 1000-1500lx, and culturing for 25-30 days for 12 hours/day.
4. Use of squalene according to claim 1 in the subculture of Mo Langen like stems, wherein in the preparation of the subculture medium, squalene is dissolved in ethyl acetate and is added after filtration sterilization and after the subculture medium is cooled to below 50 ℃.
5. The use of squalene according to claim 1 in the subculture of Mo Langen like stems, wherein the Mo Langen like stems are root like stems of the cymbidium sinense variety 'penguin Bai Momo blue' with a length of 1.0-2.0 cm and stem tips removed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310448566.XA CN116472961A (en) | 2023-04-24 | 2023-04-24 | Application of squalene in Mo Langen-like stem subculture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310448566.XA CN116472961A (en) | 2023-04-24 | 2023-04-24 | Application of squalene in Mo Langen-like stem subculture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116472961A true CN116472961A (en) | 2023-07-25 |
Family
ID=87213372
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310448566.XA Pending CN116472961A (en) | 2023-04-24 | 2023-04-24 | Application of squalene in Mo Langen-like stem subculture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116472961A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116472962A (en) * | 2023-04-24 | 2023-07-25 | 玉林师范学院 | Application of lycopene in Mo Langen-like stem proliferation culture |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107873525A (en) * | 2017-12-28 | 2018-04-06 | 武汉生物工程学院 | A kind of Chinese cymbidium quick breeding method for tissue culture |
| CN110140628A (en) * | 2019-06-28 | 2019-08-20 | 广西乙木农业科技有限公司 | A kind of high temperature and humidity rapid propagation cultivation method of Chinese cymbidium |
| CN110301355A (en) * | 2019-07-31 | 2019-10-08 | 三明市农业科学研究院 | A kind of Chinese cymbidium method for tissue culture |
| CN116420622A (en) * | 2023-04-24 | 2023-07-14 | 玉林师范学院 | Method for promoting proliferation of cymbidium sinense |
| CN116472962A (en) * | 2023-04-24 | 2023-07-25 | 玉林师范学院 | Application of lycopene in Mo Langen-like stem proliferation culture |
-
2023
- 2023-04-24 CN CN202310448566.XA patent/CN116472961A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107873525A (en) * | 2017-12-28 | 2018-04-06 | 武汉生物工程学院 | A kind of Chinese cymbidium quick breeding method for tissue culture |
| CN110140628A (en) * | 2019-06-28 | 2019-08-20 | 广西乙木农业科技有限公司 | A kind of high temperature and humidity rapid propagation cultivation method of Chinese cymbidium |
| CN110301355A (en) * | 2019-07-31 | 2019-10-08 | 三明市农业科学研究院 | A kind of Chinese cymbidium method for tissue culture |
| CN116420622A (en) * | 2023-04-24 | 2023-07-14 | 玉林师范学院 | Method for promoting proliferation of cymbidium sinense |
| CN116472962A (en) * | 2023-04-24 | 2023-07-25 | 玉林师范学院 | Application of lycopene in Mo Langen-like stem proliferation culture |
Non-Patent Citations (1)
| Title |
|---|
| 施福军等: "墨兰的无菌播种及根状茎的增殖研究", 《安徽农业科学》, vol. 36, no. 32, pages 13968 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116472962A (en) * | 2023-04-24 | 2023-07-25 | 玉林师范学院 | Application of lycopene in Mo Langen-like stem proliferation culture |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gamborg et al. | Organogenesis in callus from shoot apices of Pisum sativum | |
| US4672035A (en) | Controlled regeneration of cotton plants from tissue culture | |
| Kartha et al. | Regeneration of pea (Pisum sativum L.) plants from shoot apical meristems | |
| US20100255585A1 (en) | Plant stem cell line derived from quiescent center and method for isolating the same | |
| CN116472961A (en) | Application of squalene in Mo Langen-like stem subculture | |
| CN105475137A (en) | Method for tissue culture taking lotus reproduction and rooting into account | |
| Bais et al. | Influence of polyamines on growth and formation of secondary metabolites in hairy root cultures of Beta vulgaris and Tagetes patula | |
| TAKAYAMA et al. | Effect of Cullural Conditions on the Growth of Agrostemma githago Cells in Suspension Culture and the Concomitant Production of an Anti‐Plant Virus Substance | |
| Al-Mazrooei et al. | Optimisation of somatic embryogenesis in fourteen cultivars of sweet potato [Ipomoea batatas (L.) Lam.] | |
| CN102860257B (en) | Houttuynia cordata aerial stem tissue culture rapid propagation method | |
| Goforth et al. | The development of isolated roots of Comptonia peregrina (Myricaceae) in culture | |
| Washida et al. | Ginsenosides in hairy roots of a Panax hybrid | |
| Maldonado-Mendoza et al. | Establishment and characterization of photosynthetic hairy root cultures of Datura stramonium | |
| CN116602208A (en) | Method for regenerating cymbidium sinense rhizome plants | |
| CN116472962A (en) | Application of lycopene in Mo Langen-like stem proliferation culture | |
| Mitsuoka et al. | Effect of intracellular 2, 4-D concentration on plantlet regeneration of rice (Oryza sauva L.) callus | |
| CN116569841A (en) | Application of small molecular organic compound Prenol in Mo Langen-like stem root differentiation culture | |
| CN116439134A (en) | Application of beta-farnesene in Mo Langen-shaped stem bud differentiation culture | |
| CN103548695A (en) | Tissue culture and rapid propagation method for corydalis saxicola bunting | |
| CN116420621A (en) | Method for promoting differentiation of cymbidium sinense rhizome buds | |
| Habas et al. | In vitro multiple shoot regeneration from Petunia hybrida | |
| CN102952823A (en) | Wheat genetic transformation method | |
| GU | Callus culture of sainfoin (Onobrychis viciifolia) and plant regeneration through somatic embryogenesis | |
| CN102144550B (en) | Tissue culture method for flower buds of hemerocallis middendorfii 'prunus lannesiana wils' | |
| CN112772418A (en) | Method for tissue culture and rapid propagation of dendrobium nobile flower buds |
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 |