CN101485291B - Tissue culture multiplication method for Pinus densiflora - Google Patents
Tissue culture multiplication method for Pinus densiflora Download PDFInfo
- Publication number
- CN101485291B CN101485291B CN2008100192701A CN200810019270A CN101485291B CN 101485291 B CN101485291 B CN 101485291B CN 2008100192701 A CN2008100192701 A CN 2008100192701A CN 200810019270 A CN200810019270 A CN 200810019270A CN 101485291 B CN101485291 B CN 101485291B
- Authority
- CN
- China
- Prior art keywords
- bud
- naa
- seedlings
- concentration
- medium
- 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.)
- Expired - Fee Related
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a red pine tissue propagation method. The method comprises the following steps: taking a red pine asepsis seedling cotyledonary node (containing a cotyledon, a budding epicotyl and a 6 to 8mm hypocotyl) as an explant which is inoculated in a GD+6-BA3.0 to 5.0mg/L+NAA0.1 to 0.3mg/L culture medium so as to induce a cotyledon base to generate clustered seedlings; after 4 to 5 weeks, shifting the explant together with the differentiated clustered seedlings into a culture medium with added active carbon or NAA so as to promote the elongation of the seedlings; and when the clustered seedlings grow to 1.5 to 2.0cm, cutting off the seedlings separately, cutting the seedlings into 5 to 8mm small sections, and inoculating the small sections in a GD+1.5 to 2.5mg/L of 6-BA+0.25mg/L of NAA culture medium for further inducing the propagation of clustered seedlings. The propagated clustered seedlings can be used for a new cycle of propagation again after culture and elongation. The method can realize the clustered seedlings propagation coefficient of between 3.5 and 4.6; and a seed can propagate 20 to 30 thousand tissue culture seedlings for rootage 2 years later.
Description
Technical field
The invention belongs to biological technical field, specifically be meant Japanese red pine tissue culture enrichment procedure.
Background technology
Pinus (Pinus) is that maximum also is a most important genus in each genus of coniferous tree, nearly 116 kinds and mutation and hybrid, distribute, plant very extensive in the world, not only outstanding contributions are arranged, and in the ecosystem, have important status at aspects such as world's timber, rosin and papermaking.Owing to be subjected to the harm of various damage by disease and insect, all there are every year a large amount of pine trees withered, wherein, be subjected to pine nematode (Bursaphelenchus xylophilus) harm the most serious.This disease has caused the Japanese pine forest gross area 2,660,000 hm at present
2In 450,000 hm
2Morbidity is lost very heavy.In China, because of pine nematode cause withered pine tree add up to 5,000 ten thousand surplus strain, 2,500,000,000 yuan of direct economic losses, indirect economic loss reaches 25,000,000,000 yuan, and also with annual 6000hm
2VELOCITY DIFFUSION spread, production of forestry, the forest resources and ecotope have been caused serious destruction.
Japanese red pine is a kind of Pinus, is distributed in China East China and northern coastal area and Japan, Korea, the Soviet Union.Usually planted for construction timber, slurrying with material or shade tree, street tree, tide water protection forest, amenity forest.One of seeds that these seeds are the easiest infection pine nematodes.In Japan, owing to be subjected to the serious harm of pine wood nematode, Japanese red pine is withered in a large number.In China, Korea S, Japanese red pine is subjected to the serious threat of pine nematode equally.In order to save indigenous tree species in imminent danger, Japanese forest workers is by having set up the disease-resistant seed orchard of Japanese red pine in modes such as the disease-resistant individual plants of the serious spot seed selection of disease.Yet disease-resistant seed production is very limited, is difficult to satisfy the needs of extensive production of forestry.For this reason, press for the effective way of seeking to breed fast, in a large number existing disease-resistant material.
Plant Tissue Breeding is as a kind of alternative asexual reproduction method, for example obtained using widely in the stock breeding of eucalyptus at crops, flowers and the quick growing species of trees, also is proved to be feasible in many coniferous tree.According to the preliminary statistics, up to the present, the various countries scholar has carried out tissue culture to more than 60 kind of Pinus or mutation, hybrid, and nearly 40 have obtained the regeneration plant, wherein Chinese scholar Shen Xi ring, Huang Jianqiu etc. study 15 kinds of Pinus, and 8 kinds have obtained regeneration plant.Although the tissue culture of Pinus seeds extremely various countries scholar is paid attention to, but because difficulty is bigger, have in the existing method for tissue culture that growth coefficient is low, vitrification phenomenon is serious, be difficult to problem such as long-term subculture cultivation, make numerous researchs only stay in finishing of plant regeneration process, and to the long-term subculture of middle brood body, breed rare research in a large number, cause the seeds phoenix feathers and unicorn horns that to realize that industrialization is produced.
The report of relevant Japanese red pine tissue culture is less.Isikawa (1987) has studied the organ differentiation situation of the Japanese red pine hypocotyl and the seedling tip of a root, only finds to have the formation of adventive root on plumular axis; Choi etc. (1993) study Japanese red pine seedling organ, obtain a small amount of whole plant; (2006) such as Maruyama etc. (2005), Shoji induce somatic embryo with immature embryo respectively, and obtain regeneration plant, but have problems such as embryo callus subculture inductivity, body embryo conversion ratio are extremely low.At present domestic as yet not relevant for the morphogenetic report of Japanese red pine tissue culture.
Summary of the invention
Goal of the invention: it is precious and rare and Japanese red pine tissue culture growth coefficient is low, expand hard to tackle problem in a large number that the present invention is primarily aimed at anti-pine nematode Japanese red pine mature seed, and a kind of Japanese red pine tissue culture enrichment procedure of research invention is to satisfy need of social production.
Technical scheme: a kind of Japanese red pine tissue culture enrichment procedure may further comprise the steps:
1, the seedling cotyledon joint (epicotyl and the 6～8mm hypocotyl that contain cotyledon and just sprouted) with 3～4 weeks of Japanese red pine axenic germination is inoculated in minimal medium GD (the Gresshoffand Doy medium that contains 6-benzyl purine (6-BA) and methyl (NAA), 1972) on, the elicitor leaf base produces the bud of growing thickly;
2, explant is changed over to together with the bud of growing thickly that has broken up among the minimal medium DCR (Guptaand Durzan medium, 1985) that contains active carbon (AC) or NAA, the bud elongation promotes to grow thickly;
3, choose single the cutting down of the bud of growing thickly of robust growth, long 1.5～2.0cm, cut shoot tip meristem and be cut to the segment of growing 5～8mm, be seeded among the minimal medium GD that contains 6-BA and NAA, induce adventitious buds proliferation.
The bud of growing thickly that step 3 propagation produces changes over to and promotes its elongation in the described medium of step 2, and repeating step 3 then, move in circles, and can obtain the bud of growing thickly in a large number, a large amount of propagation of bud thereby realization is grown thickly.Step 2, step 3 subculture cycle respectively were 4～5 weeks.
More than each step condition of culture be: 23～25 ℃ of temperature, intensity of illumination 2000 Lux, illumination every day 16h.
Wherein, the concentration that produces 6-BA in the medium of the bud of growing thickly at the leaf base of elicitor described in the step 1 is 3.0～5.0mg/L, and the concentration of NAA is 0.1～0.3mg/L.
When on the DCR medium of explant being gone into to contain AC together with the bud grafting of having broken up of growing thickly, the concentration of AC is 0.5～1.5g/L in step 2; When access contained on the DCR medium of NAA, the concentration of NAA was 0.1～0.3mg/L.
The concentration of 6-BA is 1.5～2.5mg/L in the medium of inducing adventitious buds proliferation described in the step 3, and the concentration of NAA is 0.25mg/L.
Beneficial effect: the present invention is an explant with the cotyledonary node of Japanese red pine aseptic seedlings, the elicitor leaf base produces the bud of growing thickly, and after the bud of the growing thickly elongation its shoot tip meristem is extractd, and then is cut into the long segment of 5～8mm, axillalry bud is discharged under the inhibition of apical dominance, form the bud of growing thickly; Enrichment culture is cultivated the subculture mode hocket with elongation and has been overcome the vitrification phenomenon in the tissue culture procedures, makes that material can long-term subculture, thereby can breed in a large number.Compare with the indefinite bud of selecting for use explant inductions such as cotyledon, mature embryo to produce, the present invention induces that the bud of growing thickly of generation is more healthy and stronger, growth is rapider, has shortened cultivation cycle, and stabilization characteristics of genetics, is difficult for producing variation.Pass through this method, per 8～10 weeks of the bud of growing thickly can breed once, and growth coefficient remains at 3.5～4.6, and a seed can be bred after 2 years and the tissue cultivating seedling that 2～30,000 strains can be used for taking root, for extensive, short period of realizing disease-resistant material, the batch production production of high reproductive rate are laid a good foundation.
Embodiment
The following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
Embodiment 1:
Choose the anti-pine nematode Japanese red pine of introducing from Japan 1
#, 7
#The select tree mature seed, place under 4 ℃ of low temperature and handled for 4 weeks, take out with 0.1% potassium permanganate preliminary treatment 12h, flowing water flushing 1h, after flushing finishes, under aseptic condition, divest kind of a shell, be placed on 0.1% mercuric chloride through 70% ethanol surface sterilization 30s and soak 2min, behind aseptic water washing 4～5 times, move into that to cultivate into aseptic seedling in the medium standby.
Select the aseptic seedlings of sprouting for 3～4 weeks, 6～8mm place cuts hypocotyl under cotyledon, cotyledonary node (epicotyl and the 6～8mm hypocotyl that contain cotyledon and just sprouted) is as explant, be inoculated in GD+6-BA3.0～5.0mg/L+NAA0.1～0.3mg/L medium, after 4～5 weeks, explant is changed in DCR+AC 0.5～1.5g/L medium together with the bud of growing thickly that has broken up, promote the elongation of bud, use for proliferation test.
With the respectively single cutting-out of bud of growing thickly of above-mentioned length 1.5～2.0cm, be cut into 5～8mm segment after cutting shoot tip meristem, be inoculated in GD+6-BA1.5～2.5mg/L+NAA 0.25mg/L medium and induce adventitious buds proliferation; After 4～5 weeks, the bud of growing thickly that propagation is produced changes the elongation that promotes bud in DCR+AC 0.5～1.5g/L medium again over to.Carry out new round propagation after the bud of the growing thickly elongation again.
The subculture cycle of above adventitious buds proliferation, elongation was respectively for 4～5 weeks, moved in circles, per 8～10 all adventitious buds proliferations once, after 2 years, an average seed is bred tissue cultivating seedling 20000～30000 strains that to can be used for taking root.
Condition of culture: 23～25 ℃ of temperature, intensity of illumination 2000 Lux, illumination every day 16h.
Embodiment 2:
Choose the anti-pine nematode Japanese red pine of introducing from Japan 5
#, 16
#The select tree mature seed on the basis of embodiment 1, is replaced by DCR+NAA0.1～0.3mg/L with the medium of the bud elongation that promotes to grow thickly, and all the other steps are with embodiment 1, has also obtained the effect suitable with embodiment 1.
Below the validity of some concrete experiment situations prescription, explant cutting mode and the subculture mode that will help to illustrate that Japanese red pine tissue culture enrichment procedure of the present invention is used, all tests repeat 3 times at least.
The first, hormone is to the influence of Japanese red pine adventitious buds proliferation
With GD is minimal medium, adds 6-BA0.5～2.5mg/L+NAA 0.25mg/L or kinetin (KT) 0.5～2.5mg/L+NAA0.25mg/L, and relatively different hormone combinations is to the influence of adventitious buds proliferation.The result shows (table 1), and 6-BA is obvious to the grow thickly cultivation effect of bud of Japanese red pine, and in the medium that adds 6-BA1.5～2.5mg/L, growth coefficient reaches 3.5～4.6; And under 0.5～2.5mg/L concentration, KT is to the propagation nearly unavailable of the bud of growing thickly, and the explant of response is few.
Table 1 hormone is to the influence of Japanese red pine adventitious buds proliferation
| Hormone kind and concentration (mg/L) | Inoculation explant number | Response explant number (ratio %) | Growth coefficient | ||
| 6-BA | KT | NAA | |||
| 0.5 | - | 0.25 | 30 | 6(20.0) | 0.3 |
| 1.5 | - | 0.25 | 30 | 21(70.0) | 3.5 |
| 2.5 | - | 0.25 | 30 | 25(83.3) | 4.6 |
| - | 0.5 | 0.25 | 30 | 0(0) | 0 |
| - | 1.5 | 0.25 | 33 | 2(6.1) | 0.06 |
| - | 2.5 | 0.25 | 30 | 4(13.3) | 0.13 |
The second, the explant cutting mode is to the influence of Japanese red pine adventitious buds proliferation and growth
To cut into 3～8mm segment without the complete simple bud of cutting or with simple bud, and be inoculated in the GD+6-BA2.5mg/L+NAA0.25mg/L medium and breed.4～5 week back results show (table 2), the explant cutting mode has certain influence to the Japanese red pine adventitious buds proliferation: the bud of long a 1.5～2.0cm can obtain 3～4 times increment without cutting, if and be cut to 2～3 sections, then growth coefficient improves 2～3 times, cuts to such an extent that then can cause vitrifying too for a short time.
Table 2 explant cutting mode is to the influence of Japanese red pine adventitious buds proliferation and growth
| Cutting mode | The adventitious buds proliferation situation | Growing state |
| Complete simple bud | Usually breed 3～4 buds | The tip continued growth of simple bud top, newly-increased bud is thin and delicate |
| Cut into 4 section (3～4mm) | Usually breed 3～4 buds for every section, growth coefficient improves 4 times | Explant and newly-increased bud vitrifying are serious |
| Cut into 3 section (5～6mm) | Usually breed 3～4 buds for every section, growth coefficient improves 3 times | Newly-increased blastogenesis is long healthy and strong |
| Cut into 2 section (6～8mm) | Usually breed 3～4 buds for every section, growth coefficient improves 2 times | Newly-increased blastogenesis is long healthy and strong |
The 3rd, the subculture mode is to the influence of Japanese red pine adventitious buds proliferation and growth
Tissue cultivating seedling propagation normally with the continuous subculture of explant in the proliferated culture medium that contains 6-BA, this mode usually causes the vitrifying of tissue cultivating seedling, has therefore compared the influence of different subculture modes to Japanese red pine adventitious buds proliferation and growth.At first explant is inoculated among the adventitious buds proliferation medium GD+6-BA 2.5mg/L+NAA 0.25mg/L (M1) that is filtered out, be transferred to the medium (M2, M3, M4, M5) that reduces 6-BA concentration gradually after 4～5 weeks respectively or do not have any hormone but add in the medium (M6) of AC, 4～5 Zhou Houzai transfer among the proliferated culture medium M1.The result shows (table 3): the grow thickly bud vitrification phenomenon of subculture in containing higher concentration 6-BA medium is serious continuously; The bud of growing thickly that produces in the proliferated culture medium transferred in the medium that only contains NAA or AC cultivate the mode of breeding again after the elongation and overcome vitrification phenomenon, promptly M1-M5-M1 in the table 3 or M1-M6-M1 program are suitable subculture mode.
Table 3 subculture mode is to the influence of Japanese red pine adventitious buds proliferation and growth
| The subculture mode | Growth coefficient | The length of bud (mm) | The growing way of bud |
| M1-M1-M1 | 7.3 | 2～3 | 80% bud vitrifying (needle is blackish green, swelling or thin and delicate, shrinkage are curled) |
| M1-M2-M1 | 6.2 | 3～4 | 35% bud vitrifying |
| M1-M3-M1 | 6.5 | 3～4 | 15% bud vitrifying |
| M1-M4-M1 | 6.5 | 4～5 | The blastogenesis of growing thickly is long fair, the needle tubbiness |
| M1-M5-M1 | 4.3 | 5～6 | The blastogenesis of growing thickly is better long, and needle is light green tall and straight |
| M1-M6-M1 | 4.5 | 5～6 | The blastogenesis of growing thickly is long healthy and strong, and needle is light green tall and straight |
Annotate: in M2, M3, M4, the M5 medium 6-BA concentration be respectively 2.0,1.0,0.5,0mg/L, all the other are identical with M1.
In sum, a large amount of propagation of middle brood body are one of Pinus seeds most important stages of tissue culture, often become the main bottleneck of large-scale production.Long-term subculture and the propagation problem that the subculture mode that hockets has solved the Japanese red pine tissue culture cultivated in propagation prescription, explant cutting mode and enrichment culture that the present invention adopts and elongation.
Claims (1)
1. Japanese red pine tissue culture enrichment procedure is characterized in that this method may further comprise the steps:
(1) seedling cotyledon with 3～4 weeks of Japanese red pine axenic germination saves, epicotyl and 6～8mm hypocotyl that this cotyledonary node contains cotyledon and just sprouted, be inoculated on the minimal medium GD that contains 6-benzyl purine (6-BA) and methyl (NAA), the elicitor leaf base produces the bud of growing thickly; Wherein the concentration of 6-BA is 3.0～5.0mg/L, and the concentration of NAA is 0.1～0.3mg/L;
(2) explant is changed over to together with the bud of growing thickly that has broken up among the minimal medium DCR that contains active carbon (AC) or NAA, the bud elongation promotes to grow thickly; Wherein, when changing the medium that contain AC on together with the bud of growing thickly that has broken up explant, the concentration of AC is 0.5～1.5g/L; When changing the medium that contain NAA on together with the bud of growing thickly that has broken up explant, the concentration of NAA is 0.1～0.3mg/L;
(3) choose single the cutting down of the bud of growing thickly of robust growth, long 1.5～2.0cm, cut shoot tip meristem and be cut to 5～8mm segment, be seeded among the minimal medium GD that contains 6-BA and NAA, induce adventitious buds proliferation; Wherein the concentration of 6-BA is 1.5～2.5mg/L, and the concentration of NAA is 0.25mg/L;
The bud of growing thickly that step (3) propagation produces changes over to and promotes its elongation in the described medium of step (2), and then repeating step (3), moves in circles, and can obtain the bud of growing thickly in a large number, a large amount of propagation of bud thereby realization is grown thickly; The subculture cycle of step (2), step (3) was respectively for 4～5 weeks.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100192701A CN101485291B (en) | 2008-01-18 | 2008-01-18 | Tissue culture multiplication method for Pinus densiflora |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100192701A CN101485291B (en) | 2008-01-18 | 2008-01-18 | Tissue culture multiplication method for Pinus densiflora |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101485291A CN101485291A (en) | 2009-07-22 |
| CN101485291B true CN101485291B (en) | 2011-08-24 |
Family
ID=40888481
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100192701A Expired - Fee Related CN101485291B (en) | 2008-01-18 | 2008-01-18 | Tissue culture multiplication method for Pinus densiflora |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101485291B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103329803B (en) * | 2013-06-26 | 2015-04-22 | 广西壮族自治区林业科学研究院 | Tufted-bud induction and subculture method for southern pine trees |
| CN103548699B (en) * | 2013-11-22 | 2015-05-13 | 广西壮族自治区林业科学研究院 | Method for induction and proliferation culture of tissue culture buds of high fat pine tree stems |
| CN107864864B (en) * | 2017-12-25 | 2021-01-12 | 绵阳师范学院 | Tissue culture and rapid propagation method for stem segments of Kangding spruce |
| CN115316272B (en) * | 2022-08-16 | 2023-06-30 | 青岛农业大学 | Method for regenerating pinus sylvestris plants, low-temperature regeneration method and application thereof |
-
2008
- 2008-01-18 CN CN2008100192701A patent/CN101485291B/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 傅禄敏等.赤松不定根和愈伤组织与松口蘑外生菌根菌的相互作用(英文).《菌物系统》.2003,(第03期), * |
| 刘振钦等.松茸菌丝体分离培养研究初报.《吉林农业大学学报》.1989,(第02期), * |
| 王启和.联邦德国林木遗传育种研究概况.《四川林业科技》.1990,(第02期), * |
| 饶桂英等.云台林区松树死亡原因调查报告.《江苏林业科技》.1992,(第03期), * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101485291A (en) | 2009-07-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102217539B (en) | Isolated rooting culture method for fir clone | |
| CN100581352C (en) | Method for high frequency plant regenerating of tallow tree tissue culture adventitious bud | |
| CN102301952B (en) | Method for breeding chamomile | |
| CN103380730A (en) | Tissue-culture rapid propagation method for pyrus betulaefolia bunge | |
| CN101589690B (en) | Method for efficiently inducing generation of adventitious roots of Pinus densiflora tissue culture plantlets | |
| CN104885948A (en) | Method for directly regenerating plants by tea-oil tree cotyledonary nodes | |
| CN108142283B (en) | Tissue culture rapid propagation method of Acer catalpa Maxim | |
| US7932086B2 (en) | Commercially viable process for in vitro mass culture of Jatropha curcas | |
| CN101485291B (en) | Tissue culture multiplication method for Pinus densiflora | |
| CN105379624A (en) | Tissue culture fast propagation method of Eucalyptus pellita | |
| CN113207690B (en) | Efficient one-step regeneration method taking paper mulberry root as explant | |
| CN112715367B (en) | Method for carrying out Maozu secondary proliferation by using lanthanum nitrate | |
| CN104938335B (en) | The method that regeneration plant is obtained using oil tea hypocotyls | |
| CN114342804A (en) | Method for promoting regeneration of camellia oleifera bud stem plant through light control | |
| CN111990259B (en) | High-fidelity seedling breeding method for carnation | |
| CN106857251A (en) | A kind of Phoebe bournei somatic embryo and adventitious bud inducing method | |
| CN115633640B (en) | Method for regenerating adventitious buds of canna test-tube plantlet leaf sheath and application thereof | |
| CN104094848A (en) | Induction of tung tree hypocotyls callus and method for efficiently regenerating plants | |
| CN103749296A (en) | Cultivation method for promoting auxiliary bud multiplication and rooting of acacia melanoxylon plus tree | |
| CN114431154B (en) | Method for asexual propagation through acer nikoense dormant buds | |
| CN104322369B (en) | A kind of method of lombardy poplar stem segment tissue culture fast breeding | |
| CN114424749A (en) | Liriope spicata in-vitro rapid propagation method | |
| CN103999680B (en) | Plasma treatment willow branch produces the method for bud mutation | |
| CN103053429B (en) | Method for regenerating semen pharbitidis in vitro embryonic axis plant | |
| CN1491535A (en) | Root inductive method for microbody reproduction of Japan dahurian larch |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20090722 Assignee: NANJING SHENGXING HARMFUL BIOLOGICAL CONTROL TECHNOLOGY CO.,LTD. Assignor: Nanjing Forestry University Contract record no.: 2018320000400 Denomination of invention: Tissue Culture and Proliferation of Pinus densiflora Granted publication date: 20110824 License type: Common License Record date: 20181218 Application publication date: 20090722 Assignee: Nanjing Forest Protection Bird Biotechnology Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000399 Denomination of invention: Tissue Culture and Proliferation of Pinus densiflora Granted publication date: 20110824 License type: Common License Record date: 20181218 Application publication date: 20090722 Assignee: Guizhou Linxin Ecological Agriculture Technology Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000401 Denomination of invention: Tissue Culture and Proliferation of Pinus densiflora Granted publication date: 20110824 License type: Common License Record date: 20181218 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110824 Termination date: 20220118 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |