CN102217539B - Isolated rooting culture method for fir clone - Google Patents
Isolated rooting culture method for fir clone Download PDFInfo
- Publication number
- CN102217539B CN102217539B CN201110129388A CN201110129388A CN102217539B CN 102217539 B CN102217539 B CN 102217539B CN 201110129388 A CN201110129388 A CN 201110129388A CN 201110129388 A CN201110129388 A CN 201110129388A CN 102217539 B CN102217539 B CN 102217539B
- Authority
- CN
- China
- Prior art keywords
- culture
- medium
- root
- rooting
- fir
- 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
- 238000012136 culture method Methods 0.000 title abstract description 5
- 238000000034 method Methods 0.000 claims abstract description 18
- 239000000463 material Substances 0.000 claims abstract description 7
- 239000001963 growth medium Substances 0.000 claims abstract description 6
- 239000003415 peat Substances 0.000 claims abstract description 4
- 239000002689 soil Substances 0.000 claims abstract description 4
- 239000002609 medium Substances 0.000 claims description 61
- 244000050510 Cunninghamia lanceolata Species 0.000 claims description 39
- 239000005648 plant growth regulator Substances 0.000 claims description 31
- 230000006698 induction Effects 0.000 claims description 16
- 230000035755 proliferation Effects 0.000 claims description 8
- 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 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- 239000005720 sucrose Substances 0.000 claims description 6
- 230000008859 change Effects 0.000 claims description 5
- 238000002054 transplantation Methods 0.000 claims description 4
- 239000006870 ms-medium Substances 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 abstract description 8
- 238000011161 development Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 239000011159 matrix material Substances 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 abstract description 3
- 230000001939 inductive effect Effects 0.000 abstract description 2
- 241000234282 Allium Species 0.000 abstract 1
- 235000002732 Allium cepa var. cepa Nutrition 0.000 abstract 1
- 239000012883 rooting culture medium Substances 0.000 abstract 1
- 230000012010 growth Effects 0.000 description 13
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 9
- 238000011081 inoculation Methods 0.000 description 8
- 229910052799 carbon Inorganic materials 0.000 description 7
- 230000000694 effects Effects 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 238000012549 training Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000218691 Cupressaceae Species 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000218631 Coniferophyta Species 0.000 description 1
- 241000425037 Toona sinensis Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- -1 cover Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000003630 growth substance Substances 0.000 description 1
- 230000003648 hair appearance Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015816 nutrient absorption Nutrition 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
- 238000003466 welding Methods 0.000 description 1
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses an isolated rooting culture method for a fir clone. The method comprises the following steps of: selecting basal sprouts of a trunk of an original plant of the excellent fir clone as a tissue culture material, inoculating the basal sprouts to a successive multiplication culture medium to perform successive multiplication culture, inducing and differentiating adventitious buds, and transferring the adventitious buds to an MS (Murashige and Skoog) culture medium to perform elongation culture; when the adventitious buds are grown to 2 to 3 centimeters, transferring the adventitious buds to an induced rooting culture medium, and performing culture to obtain rooted test tube plantlets; and transplanting the rooted test tube plantlets, and mixing loess and peat soil in a ratio of 3:1 to form s transplanting matrix. Compared with a fir tissue culture method in the prior art, the isolated rooting culture method for the fir clone has the advantages that: the rooting difficulty problem of tissue culture plantlets of different excellent clones of fir under the tissue culture condition is effectively solved, a stable and efficient tissue culture system of the fir is established, the highest rooting rate can reach 100 percent (onion 020), and a technical support is provided for quickening the industrialized development of the fir tissue culture plantlets.
Description
Technical field
The present invention relates to the China fir vegetative propagation technique, be specifically related to the stripped culture of rootage method of a kind of China fir clone.
Background technology
China fir (Cunninghamia lanceolata) is the evergreen megaphanerophyte of gymnosperm Taxodiaceae (Taxodiaceae), property happiness warm and moist weather, and growth is rapid, and trunk is perfectly straight, up to 30m, the diameter of a cross-section of a tree trunk 1.3 meters above the ground 2.5～3m.The China fir material is light and soft, careful, texture is straight, is prone to processing; Supply usefulness such as building, bridge, shipbuilding, electric welding, mine timber, wooden Chinese toon, and can make raw materials such as papermaking, weaving; Bark and root, leaf are used as medicine, and can dispel pathogenic wind and remove dampness, astringing to arrest bleeding; The seed oil-containing is about 20%, supplies soap system.
China fir is China's main reproducting tree species in south and most important commodity commerical tree species, and cultivation history is long, and its afforestation area and stand all occupy the first place of China forest plantation.Along with the improving constantly of management level, breeding obtains paying much attention to strong sprout in recent years.South China fir producing region seed selection a large amount of excellent Chinese fir clones, and utilize the tissue culture rapid propagating technology that these choiceness have been carried out asexual grow seedling afforestation, obtained significant effect of increasing production.But because different excellent Chinese fir clones are in the otherness of character inheritance and physiological foundation of taking root; Cause different China fir clones under same group of training prescription condition success rate and efficient aspect have very big difference, greatly influenced good China fir clonal tissue culture seedling and cultivated and apply.
Summary of the invention
Goal of the invention: to the deficiency that exists in the prior art; The purpose of this invention is to provide the stripped culture of rootage method of a kind of China fir clone; And then the difficult problem of taking root of the different choiceness of solution China fir tissue cultivating seedling under conditions of tissue culture; The group training system of China fir stability and high efficiency is set up in realization, for accelerating the industrialized development of China fir tissue cultivating seedling technical support is provided.
Technical scheme: in order to realize the foregoing invention purpose, the technical scheme that the present invention adopts is following:
A kind of China fir clone culture of rootage method that exsomatizes may further comprise the steps:
(1) the base portion coppice shoot of choosing the trunk of excellent Chinese fir ortet is the tissue culture material, inserts the shoot proliferation medium and carries out shoot proliferation and cultivate, and induces to differentiate indefinite bud, indefinite bud is changed on the MS medium extend cultivation then; The shoot proliferation culture medium prescription is 3/4MS+6BA0.3mgL
-1+ NAA0.02mgL
-1+ sucrose 3%;
When (2) indefinite bud of step (1) is elongated to 2～3cm, change the root induction medium culture over to, get rooting tube plantlet; The root induction medium is a minimal medium with 1/2MS or 1/4MS, adds plant growth regulator and 2% sucrose therein; Plant growth regulator is IBA0.05 ~ 0.4mgL
-1, NAA0.05 ~ 0.4mgL
-1, IAA0.1 ~ 1.0mgL
-1, or IBA0.05 ~ 0.4mgL
-1With NAA0.05 ~ 0.4mgL
-1Mixing;
(3) transplantation rooting test-tube plantlet is selected for use after loess, the peat soil 3:1 mixed as transplanting medium.
Preferably: in the step (2), the root induction medium is 1/2MS, and plant growth regulator is IBA0.1 ~ 0.4mgL
-1Or NAA0.1 ~ 0.2mgL
-1
Preferably: in rapid (2), the root induction medium is 1/4MS, and plant growth regulator is IBA0.1 ~ 0.2mgL
-1Or NAA0.05 ~ 0.2mgL
-1
Preferably: in the step (2), the root induction medium is 1/2MS, and plant growth regulator is IBA0.05 ~ 0.2mgL
-1With NAA0.05 ~ 0.4mgL
-1Mixing.
Preferably: in the step (2), the root induction medium is 1/4MS, and plant growth regulator is IBA0.05 ~ 0.2mgL
-1With NAA0.05 ~ 0.4mgL
-1Mixing.
Preferably: in the step (3),, rooting tube plantlet put under the natural lighting refine seedling, unclamp blake bottle day by day and seal film until removing at last in transplantation rooting test-tube plantlet the last week.
Preferably: in the step (3), the root length of test-tube plantlet is not less than 1cm.
Preferably: in the step (3), the root length of test-tube plantlet is 1～3cm.
In group training process, rooting of vitro seedling receives influence of various factors such as medium, hormone and condition of culture.Generally speaking, rooting of vitro seedling all need carry out in low salt culture medium.This is cause medium osmotic pressure to change because salinity changes, thus influence test-tube plantlet to nutrient absorption and in medium h substance.And the variation of salinity also makes, and nitrogen concentration drops to a favourable level that is fit to take root in the plant body.Among the application; Test-tube plantlet rooting efficiency on the medium of 1/4MS in ocean 062, ocean 061, ocean 021, ocean 023 is better; Its rooting rate is the highest can to reach 93.3%, 56.7%, 70%, 90% respectively; But the test-tube plantlet in ocean 020 is taken root on the minimal medium of 1/2MS better, and the highest rooting rate is 100%.
The test-tube seedling transplanting survival rate just is to identify the most effectively foundation of root of hair quality and root of hair technology; The transplanting survival rate of the application's China fir test-tube plantlet reaches more than 70%, can explain that the China fir test-tube plantlet adventive root that the stripped culture of rootage method of China fir clone is cultivated is high-quality.
Beneficial effect: compare with China fir tissue culture method of the prior art; The stripped culture of rootage method advantage of China fir clone of the present invention is: the difficult problem of taking root that efficiently solves the different choiceness of China fir tissue cultivating seedling under conditions of tissue culture; The group training system of China fir stability and high efficiency is set up in realization, and rooting rate can reach 100% (ocean 020), for accelerating the industrialized development of China fir tissue cultivating seedling technical support is provided; Can produce good social benefit, have good economic outlook.
Description of drawings
Fig. 1 is the figure of taking root in ocean 062;
Fig. 2 is the figure of taking root in ocean 020;
Fig. 3 is the figure of taking root in ocean 061;
Fig. 4 is the figure of taking root in ocean 021;
Fig. 5 is the figure of taking root in ocean 023;
Fig. 6 is that test-tube seedling transplanting becomes figure alive.
Embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is done further explanation.
Following examples adopt Yang Kou forest farm, the Fujian excellent Chinese fir clone aseptic seedling of Nanjing Forestry University and Fujian cooperation seed selection, and the application abbreviates each sample respectively as: ocean 062, ocean 020, ocean 061, ocean 023 and ocean 021.These materials originate from the defect individual in the superior families that the second generation genetic improvement of Nan Lin-Fujian cooperation development selects in the works, have carried out the clonal test in more than 10 year after breeding into clone, select the back as the choiceness accelerates multiplication again.All tissue culture materials all originate from the base portion coppice shoot of the trunk of ortet.
Below the condition of culture (removing special indicating) that uses of each embodiment be: all additional agar 0.6% in the medium, pH5.8, illumination (2000lx) 14h/d, about 25 ℃ of cultivation temperature.
Embodiment 1The differentiation of bud and elongation
Each tissue culture material (ocean 062, ocean 020, ocean 061, ocean 023 and ocean 021) is put into the shoot proliferation medium respectively cultivate, induce to differentiate indefinite bud, the shoot proliferation culture medium prescription adopts 3/4MS+6BA0.3 mgL
-1+ NAA0.02mgL
-1+ sucrose 3%.To change over to through the indefinite bud of inducing differentiation on the MS medium of no any growth regulator and extend cultivation.
Embodiment 2Root induction
When each indefinite bud length to 2 of embodiment 1～3cm, change it over to root media and cultivate it and take root.In root media, add plant growth regulator and sucrose 2%, observe statistical data.The data statistics index comprises: the induction frequency of adventive root (%)=grow indefinite bud number/inoculation indefinite bud number * 100% of adventive root; The sum of average indefinite radical=adventive root/the induce indefinite bud number of adventive root; The induction frequency of formation ability (the RFC)=average indefinite radical * adventive root of adventive root.
In cultivated in ocean 062, the experimental result of the situation of choosing of root media and plant growth regulator and correspondence thereof was as shown in table 1, and is visible from table, and it is best to take root with the medium of additional NAA, when NAA concentration<0.2mgL
-1The time, rooting rate becomes positive correlation with NAA concentration; As NAA Nong Du>0.2mgL
-1The time, be negative correlation; Equal 0.2mgL and work as NAA concentration
-1The time, rooting rate is up to 46.7%.The medium effect of additional IAA is the poorest.Under the NAA of same concentrations level, rooting rate is apparently higher than the 1/2MS medium in the 1/4MS medium; Under the IBA of same concentrations level, rooting rate is also apparently higher than the 1/2MS medium in the 1/4MS medium.Wherein with the additional NAA0.2mgL of 1/4MS
-1The medium effect is best, and rooting rate is 56.7%, and explant grows fine.
The situation of choosing of foreign 062 root media of table 1 and plant growth regulator and corresponding experimental result table thereof
| Medium and plant growth regulator | Inoculation explant number | Rooting rate | Mean elements (bar) | Adventive root forms ability |
| 1/2MS+NAA 0.05 mg·L -1 | 30 | 20 | 2.5 | 0.5 |
| 1/2MS+NAA 0.1 mg·L -1 | 30 | 30 | 5.5 | 1.65 |
| 1/2MS+NAA 0.2 mg·L -1 | 30 | 46.7 | 4 | 1.87 |
| 1/2MS+NAA 0.4 mg·L -1 | 30 | 6.7 | 1.5 | 0.10 |
| 1/2MS+IBA 0.05 mg·L -1 | 30 | 3.3 | 1 | 0.03 |
| 1/2MS+IBA 0.1 mg·L -1 | 30 | 10 | 1.3 | 0.13 |
| 1/2MS+IBA 0.2 mg·L -1 | 30 | 13.3 | 2 | 0.26 |
| 1/2MS+IBA 0.4 mg·L -1 | 30 | 6.7 | 1.5 | 0.10 |
| 1/2MS+IAA 0.1 mg·L -1 | 30 | 0 | 0 | 0 |
| 1/2MS+IAA 0.2 mg·L -1 | 30 | 0 | 0 | 0 |
| 1/2MS+IAA 0.5 mg·L -1 | 30 | 26.7 | 3.5 | 0.93 |
| 1/2MS+IAA 1.0 mg·L -1 | 30 | 0 | 0 | 0 |
| 1/4MS+NAA0.05 mg·L -1 | 30 | 43.3 | 3.1 | 1.34 |
| 1/4MS+NAA0.1 mg·L -1 | 30 | 50 | 3.9 | 1.95 |
| 1/4MS+NAA0.2 mg·L -1 | 30 | 56.7 | 4 | 2.27 |
| 1/4MS+NAA0.4 mg·L -1 | 30 | 13.3 | 2 | 0.26 |
| 1/4MS+IBA0.05 mg·L -1 | 30 | 3.3 | 2 | 0.06 |
| 1/4MS+IBA0.1 mg·L -1 | 30 | 13.3 | 1.8 | 0.24 |
| 1/4MS+IBA0.2 mg·L -1 | 30 | 20.0 | 2.3 | 0.46 |
| 1/4MS+IBA0.4 mg·L -1 | 30 | 10 | 1.7 | 0.17 |
| 1/4MS+IAA 0.1 mg·L -1 | 30 | 0 | 0 | 0 |
| 1/4MS+IAA 0.2 mg·L -1 | 30 | 13.3 | 1.8 | 0.24 |
| 1/4MS+IAA 0. 5 mg·L -1 | 30 | 33.3 | 2.7 | 0.90. |
In cultivate in ocean 062; The experimental result of the situation of choosing of root media and plant growth regulator (2 kinds) and correspondence thereof is as shown in table 2, and is visible from table, when in the 1/2MS medium, adding NAA and IBA simultaneously; China fir adventitious bud rooting rate generally rises, and average rooting rate is 53.9%.Be grown in additional 0.1mgL
-1NAA and 0.4 mgL
-1The rooting rate of the tissue cultivating seedling on the medium of IBA can reach 73.3%, has improved the tissue cultivating seedling rooting rate greatly, and along with the prolongation of time, rooting rate also rises to some extent, and the explant growth conditions is good.Hence one can see that, and NAA and IBA synergy more help the China fir adventitious bud rooting.After indefinite bud inserts root media 20d, there is adventive root to grow on the root media of additional IBA of 1/2MS and NAA, adventive root length to 1 ~ 1.5cm behind the 23d.On the root media of additional separately IBA of 1/2MS, 1/4MS and NAA, the root induction time is wanted 3 ~ 5d in evening, and this explanation NAA and IBA are used, and can obviously shorten rootage duration.
Under the same hormone combination, in the 1/4MS medium, the average rooting rate of explant reaches 82.2%, 53.9% average rooting rate under the 1/2MS medium.At additional 0.05mgL
-1NAA and 0.05 mgL
-1The explant rooting rate is the highest on the medium of IBA, is 93.3%; At additional 0.1mgL
-1NAA and 0.2 mgL
-1Rooting rate is 90% on the medium of IBA, and it is higher that adventive root forms ability, and the plant strain growth state is better than the former.In NAA concentration is 0.05 mgL
-1Average rooting rate is 84.2%; NAA concentration is 0.1mgL
-1The Shi Pingjun rooting rate is 85%, and is in rising trend with the increase rooting rate of IBA concentration, reaches the highest during IBA0.2; NAA concentration is 0.2 mgL
-1The Shi Pingjun rooting rate is 77.5%, when IBA0.2, reaches the highest.Hence one can see that, and the best in ocean 062 is taken root for filling a prescription and is 1/4MS+NAA0.1mgL
-1+ IBA0.2mgL
-1, the figure of taking root is as shown in Figure 1.
The situation of choosing and the experimental result of foreign 062 root media of table 2 and plant growth regulator (2 kinds)
| Medium and plant growth regulator | Inoculation explant number | Rooting rate | Mean elements (bar) | Adventive root forms ability |
| 1/2MS+NAA0.05mg·L -1+IBA0.05mg·L -1 | 30 | 43.3 | 2.4 | 1.04 |
| 1/2MS+NAA0.05mg·L -1+IBA0.1mg·L -1 | 30 | 53.3 | 1.7 | 0.91 |
| 1/2MS+NAA0.05mg·L -1+IBA0.2 mg·L -1 | 30 | 60 | 2.6 | 1.59 |
| 1/2MS+NAA0.05mg·L -1+IBA0.4mg·L -1 | 30 | 56.7 | 2.7 | 1.53 |
| 1/2MS+NAA0.1mg·L -1+IBA0.05mg·L -1 | 30 | 56.7 | 3.8 | 2.15 |
| 1/2MS+NAA0.1mg·L -1+IBA0.1mg·L -1 | 30 | 40 | 1.9 | 0.76 |
| 1/2MS+NAA0.1mg·L -1+IBA0.2mg·L -1 | 30 | 56.7 | 2.8 | 1.59 |
| 1/2MS+NAA0.1mg·L -1+IBA0.4mg·L -1 | 30 | 73.3 | 2.5 | 1.83 |
| 1/2MS+NAA0.2mg·L -1+IBA0.05mg·L -1 | 30 | 33.3 | 3.2 | 1.07 |
| 1/2MS+NAA0.2mg·L -1+IBA0.1mg·L -1 | 30 | 50 | 4.9 | 2.45 |
| 1/2MS+ NAA0.2mg·L -1+ IBA 0.2mg·L -1 | 30 | 66.7 | 2.8 | 1.87 |
| 1/2MS+NAA0.2mg·L -1+IBA0.4mg·L -1 | 30 | 56.7 | 1.9 | 1.07 |
| 1/4MS+NAA0.05mg·L -1+IBA0.05mg·L -1 | 30 | 93.3 | 2.3 | 2.15 |
| 1/4MS+NAA0.05mg·L -1+IBA0.1mg·L -1 | 30 | 80 | 2 | 1.6 |
| 1/4MS+NAA0.05mg·L -1+IBA0.2 mg·L -1 | 30 | 80 | 3.3 | 2.64 |
| 1/4MS+NAA0.05mg·L -1+IBA0.4mg·L -1 | 30 | 83.3 | 3.4 | 2.83 |
| 1/4MS+NAA0.1mg·L -1+IBA0.05mg·L -1 | 30 | 80 | 2.8 | 2.24 |
| 1/4MS+NAA0.1mg·L -1+IBA0.1mg·L -1 | 30 | 83.3 | 2.5 | 2.08 |
| 1/4MS+NAA0.1mg·L -1+IBA0.2mg·L -1 | 30 | 90 | 2.8 | 2.52 |
| 1/4MS+NAA0.1mg·L -1+IBA0.4mg·L -1 | 30 | 86.7 | 2.6 | 2.25 |
| 1/4MS+NAA0.2mg·L -1+IBA0.05mg·L -1 | 30 | 80 | 2.7 | 2.16 |
| 1/4MS+NAA0.2mg·L -1+IBA0.1mg·L -1 | 30 | 73.3 | 2.4 | 1.76 |
| 1/4MS+NAA0.2mg·L -1+IBA0.2mg·L -1 | 30 | 86.7 | 2.9 | 2.51 |
| 1/4MS+NAA0.2mg·L -1+IBA0.4 mg·L -1 | 30 | 70 | 3 | 2.1 |
In cultivate in ocean 020; The situation of choosing of root media and plant growth regulator and corresponding experimental result thereof are as shown in table 3, and be visible from table 2, best with the situation of taking root of explant in the medium of additional IAA; Average rooting rate is 95%, and rhizome junction callus is few; At IAA1.0 mgL
-1Rooting rate reaches 100%, and the bar number of adventive root is many, and the formation ability of adventive root is the highest, is 8.6.The average rooting rate of explant is 70% in the medium of additional NAA, is 0.2 mgL in concentration
-1With 0.4 mgL
-1Two prescriptions in the explant base portion expand, callus is many, the root that bears is thinner and more delicate.At the medium of additional IBA, average rooting rate is merely 25.8%.Hence one can see that, and the best root media of foreign 020 genotype is: 1/2MS+IAA 1.0mgL
-1The indefinite bud that ocean 020 is induced changes 1/4MS+NAA0.1mgL over to
-1+ IBA0.2mgL
-1In, rooting rate is 100%, 5.3 of mean elements, and the seedling growth conditions is good, and is as shown in Figure 2.
The situation of choosing of foreign 020 root media of table 3 and plant growth regulator and corresponding experimental result thereof
| Medium and plant growth regulator | Inoculation explant number | Rooting rate | Mean elements (bar) | Adventive root forms ability |
| 1/2MS+NAA 0.05mg·L -1 | 30 | 70 | 5.6 | 3.92 |
| 1/2MS+NAA 0.1 mg·L -1 | 30 | 80 | 2.7 | 2.16 |
| 1/2MS+NAA 0.2 mg·L -1 | 30 | 66.7 | 4.5 | 3.00 |
| 1/2MS+NAA 0.4 mg·L -1 | 30 | 63.3 | 5.4 | 3.41 |
| 1/2MS+IBA 0.05mg·L -1 | 30 | 13.3 | 2.3 | 0.31 |
| 1/2MS+IBA 0.1 mg·L -1 | 30 | 13.3 | 2.3 | 0.31 |
| 1/2MS+IBA 0.2 mg·L -1 | 30 | 40 | 3 | 1.2 |
| 1/2MS+IBA 0.4 mg·L -1 | 30 | 36.7 | 1.6 | 0.59 |
| 1/2MS+IAA 0.1 mg·L -1 | 30 | 86.7 | 5.8 | 5.0 |
| 1/2MS+IAA 0.2 mg·L -1 | 30 | 93.3 | 4.3 | 4.0 |
| 1/2MS+IAA 0.5mg·L -1 | 30 | 100 | 4.5 | 4.5 |
| 1/2MS+IAA 1.0mg·L -1 | 30 | 100 | 8.6 | 8.6 |
In cultivate in ocean 061; The experimental result of the situation of choosing of root media and plant growth regulator and correspondence thereof is as shown in table 4, and visible from table, foreign 061 genotype is in the minimal medium of 1/2MS; NAA, IBA, the IAA of additional variable concentrations; Its rooting rate is all lower, and the highest rooting rate is 36.7%, and average rooting rate is merely 22.0%.When in the 1/4MS medium, adding NAA and IBA simultaneously, foreign 061 genotype adventitious bud rooting rate rises to some extent, and average rooting rate reaches 50.3%.Being grown in concentration is 0.1mgL
-1NAA and 0.2 mgL
-1Adventitious bud rooting rate on the medium of IBA is up to 56.7%, and it is also the highest that adventive root forms ability, is 1.87, and the figure of specifically taking root is as shown in Figure 3.Therefore, ocean 061 genotypic the best is taken root for filling a prescription and is: 1/4MS+NAA0.1mgL
-1+ IBA0.2mgL
-1
The situation of choosing of foreign 061 root media of table 4 and plant growth regulator and corresponding experimental result thereof
| Medium and plant growth regulator | Inoculation explant number | Rooting rate | Mean elements (bar) | Adventive root forms ability |
| 1/2MS+NAA 0.05mg·L -1 | 30 | 16.7 | 1.4 | 0.23 |
| 1/2MS+ NAA 0.1 mg·L -1 | 30 | 20 | 4 | 0.8 |
| 1/2MS+ NAA 0.2 mg·L -1 | 30 | 20 | 2 | 0.4 |
| 1/2MS+ NAA 0.4 mg·L -1 | 30 | 13.3 | 2 | 0.26 |
| 1/2MS+IBA 0.05mg·L -1 | 30 | 20 | 4.1 | 0.82 |
| 1/2MS+IBA 0.1 mg·L -1 | 30 | 26.7 | 2 | 0.53 |
| 1/2MS+IBA 0.2 mg·L -1 | 30 | 20 | 5 | 1.0 |
| 1/2MS+IBA 0.4 mg·L -1 | 30 | 30 | 5 | 1.5 |
| 1/2MS+IAA 0.1 mg·L -1 | 30 | 36.7 | 3.5 | 1.28 |
| 1/2MS+IAA 0.2 mg·L -1 | 30 | 10 | 2 | 0.2 |
| 1/2MS+IAA 0.5mg·L -1 | 30 | 33.3 | 2.5 | 0.83 |
| 1/2MS+IAA 1.0mg·L -1 | 30 | 16.7 | 5.4 | 0.90 |
| 1/2MS+NAA 0.05mg·L -1 | 30 | 16.7 | 1.4 | 0.23 |
| 1/4MS+NAA0.05mg·L -1+IBA0.05mg·L -1 | 30 | 46.7 | 2.3 | 1.07 |
| 1/4MS+NAA0.05mg·L -1+IBA0.1mg·L -1 | 30 | 50 | 2 | 1.0 |
| 1/4MS+NAA0.05mg·L -1+IBA0.2 mg·L -1 | 30 | 53.3 | 2.8 | 1.49 |
| 1/4MS+NAA0.05mg·L -1+IBA0.4mg·L -1 | 30 | 50 | 3.4 | 1.7 |
| 1/4MS+NAA0.1mg·L -1+IBA0.05mg·L -1 | 30 | 50 | 2.6 | 1.3 |
| 1/4MS+NAA0.1mg·L -1+IBA0.1mg·L -1 | 30 | 53.3 | 2.5 | 1.33 |
| 1/4MS+NAA0.1mg·L -1+IBA0.2mg·L -1 | 30 | 56.7 | 3.3 | 1.87 |
| 1/4MS+NAA0.1mg·L -1+IBA0.4mg·L -1 | 30 | 56.7 | 2.8 | 1.59 |
| 1/4MS+NAA0.2mg·L -1+IBA0.05mg·L -1 | 30 | 46.7 | 2.7 | 1.26 |
| 1/4MS+NAA0.2mg·L -1+IBA0.1mg·L -1 | 30 | 43.3 | 2.4 | 1.04 |
| 1/4MS+NAA0.2mg·L -1+IBA0.2mg·L -1 | 30 | 53.3 | 2.9 | 1.55 |
| 1/4MS+NAA0.2mg·L -1+IBA0.4 mg·L -1 | 30 | 43.3 | 3 | 1.30 |
In cultivate in ocean 021; The experimental result of the root media of optimizing and the situation of choosing of plant growth regulator and correspondence thereof is as shown in table 5; Visible from table 5, during with IBA, on average rooting rate can reach 59.9% to the explant in ocean 021 at the NAA of the additional variable concentrations of 1/4MS minimal medium.When NAA concentration is 0.1 mgL
-1, IBA concentration is 0.2 mgL
-1The time adventitious bud rooting rate be 70%, mean elements is 2.8, it is the strongest that average adventive root forms ability, is 1.96, the figure of specifically taking root is as shown in Figure 4.Therefore, foreign 021 genotype the best is taken root for filling a prescription and is: 1/4MS+NAA0.1mgL
-1+ IBA0.2mgL
-1
The root media that table 5 ocean 021 is optimized and the situation of choosing of plant growth regulator and corresponding experimental result thereof
| Medium and plant growth regulator | Inoculation explant number | Rooting rate | Mean elements (bar) | Adventive root forms ability |
| 1/4MS+NAA0.1mg·L -1+IBA0.05mg·L -1 | 30 | 46 | 2.5 | 1.15 |
| 1/4MS+NAA0.1mg·L -1+IBA0.1mg·L -1 | 30 | 56.7 | 2.4 | 1.36 |
| 1/4MS+NAA0.1mg·L -1+IBA0.2mg·L -1 | 30 | 70 | 2.8 | 1.96 |
| 1/4MS+NAA0.1mg·L -1+IBA0.4 mg·L -1 | 30 | 66.7 | 2.6 | 1.73 |
In cultivate in ocean 023; The experimental result of the root media of optimizing and the situation of choosing of plant growth regulator and correspondence thereof is as shown in table 6; Visible from table 6, during with IBA, on average rooting rate reaches 81.7% to foreign 023 genotype at the NAA of the additional variable concentrations of 1/4MS minimal medium.At additional NAA0.1 mgL
-1With IBA0.2 mgL
-1Medium on, the adventitious bud rooting rate reaches 90%, mean elements 3.8, adventive root formation ability is 3.42, plant grows fine, the figure of specifically taking root is as shown in Figure 5.Therefore, ocean 023 genotypic the best is taken root for filling a prescription and is: 1/4MS+NAA0.1mgL
-1+ IBA0.2mgL
-1
The root media that table 6 ocean 023 is optimized and the situation of choosing of plant growth regulator and corresponding experimental result thereof
| Medium and plant growth regulator | Inoculation explant number | Rooting rate | Mean elements (bar) | Adventive root forms ability |
| 1/4MS+NAA0.1mg·L -1+IBA0.05mg·L -1 | 30 | 73.3 | 2.8 | 2.05 |
| 1/4MS+NAA0.1mg·L -1+IBA0.1mg·L -1 | 30 | 83.3 | 2.5 | 2.08 |
| 1/4MS+NAA0.1mg·L -1+IBA0.2mg·L -1 | 30 | 90 | 3.8 | 3.42 |
| 1/4MS+NAA0.1mg·L -1+IBA0.4 mg·L -1 | 30 | 80 | 3 | 2.4 |
Embodiment 3
Ocean 061 is inoculated in 1/4MS+NAA0.1mgL
-1+ IBA0.2mgL
-1In the medium, in medium, add variable concentrations active carbon (table 7), cultural method is observed statistics with embodiment 2 behind the 35d.After inserting the active carbon medium, the aseptic seedling growing way a little less than, when 26d, found that just adventive root grows.Can be known that by table 7 rooting rate is 46.7 on the medium of active carbon 0.02% adding, constantly descend with the increase rooting rate of concentration of activated carbon, be 0 at active carbon greater than 0.2% o'clock rooting rate; And the medium rooting rate of non-activity charcoal is 56.7%.Can know that active carbon is unfavorable for the China fir adventitious bud rooting.
The influence that table 7 active carbon is taken root to ocean 061
| Active carbon (mg/L) | Inoculation explant number | The explant number of taking root | Rooting rate (%) |
| 0.2 | 15 | 7 | 46.7 |
| 0.5 | 15 | 6 | 40 |
| 1.0 | 15 | 3 | 20 |
| 2.0 | 15 | 0 | 0 |
| 5.0 | 15 | 0 | 0 |
Embodiment 4The domestication of regeneration plant and transplanting
The rooting tube plantlet of embodiment 2 is transplanted to put under the natural lighting the last week and is refined seedling, unclamps blake bottle day by day and seals film until removing at last, in culturing room, unclamps earlier lid; Place 2d, move on to indoorly then, place 3d about 25 ℃, flush away base portion medium; Be transplanted in the matrix, cover, spray water every day 2 ~ 3 times, to keep humidity with Polypropylence Sheet; After this 7d will note the temperature and humidity that keeps certain, prevents the bad root phenomenon that occurs because of humidity is excessive simultaneously again, opens Polypropylence Sheet about 14d; Keep certain humidity, spray water every day 1 ~ 2 time, will note simultaneously letting in air.Be used as transplanting medium after selecting loess, peat soil 3:1 mixed for use.By the length of root with test-tube plantlet be divided into short root (below the 1cm), middle root (1～3cm) with three types of long root (more than the 3cm), each type is got 20 strains and is made an experiment.Plant into matrix after test-tube plantlet being taken out and cleans the medium of root, keep ventilating and 80 ~ 90% ambient humidity.Routine observation, the survival rate and the growing state of statistics test-tube plantlet behind the 30d, concrete outcome is as shown in table 8.In the table 8, the transplanting survival rate of the rooting tube plantlet that the group training obtains is more than 70%, and root length plantlet of transplant effect when 1～3cm is best; Have the new root of part to take place, plant normal growth, basal part of stem have sprouting to take out life; The proof test-tube seedling transplanting becomes to live and begins growth, and the concrete growth figure that transplants is as shown in Figure 6.
Table 8 a long table as a result that influences to transplant survival
| Handle | The root type | Handle the strain number | The strain number becomes to live | The root growth situation | The plant strain growth situation |
| 1 | <1cm | 20 | 14 | Elongation growth | Robust growth |
| 2 | 1~3cm | 20 | 17 | Elongation growth, new root takes place | Basal part of stem grows sprouting, riotous growth |
| 3 | >3cm | 20 | 12 | New root takes place, and part is rotted | Growth is normal |
Claims (8)
1. the stripped culture of rootage method of China fir clone is characterized in that, may further comprise the steps:
(1) the base portion coppice shoot of choosing the trunk of excellent Chinese fir ortet is the tissue culture material, inserts the shoot proliferation medium and carries out shoot proliferation and cultivate, and induces to differentiate indefinite bud, indefinite bud is changed on the MS medium extend cultivation then; Wherein, the shoot proliferation culture medium prescription is 3/4MS+6BA0.3mgL
-1+ NAA0.02mgL
-1+ sucrose 3%;
When (2) indefinite bud of step (1) is elongated to 2～3cm, change the root induction medium culture over to, get rooting tube plantlet; The root induction medium is a minimal medium with 1/2MS or 1/4MS, adds plant growth regulator and 2% sucrose therein; Plant growth regulator is IBA0.05～0.4mgL
-1, or be NAA0.05～0.4mgL
-1, or be IAA0.1～1.0mgL
- 1, or be IBA0.05～0.4mgL
-1With NAA0.05～0.4mgL
-1Mixing;
(3) transplantation rooting test-tube plantlet selects for use loess, peat soil mixed after to be used as transplanting medium at 3: 1.
2. the China fir clone according to claim 1 culture of rootage method that exsomatizes, it is characterized in that: in the step (2), the root induction medium is 1/2MS, and plant growth regulator is IBA0.1～0.4mgL
-1Or NAA0.1～0.2mgL
-1
3. the China fir clone according to claim 1 culture of rootage method that exsomatizes, it is characterized in that: in the step (2), the root induction medium is 1/4MS, and plant growth regulator is IBA0.1～0.2mgL
-1Or NAA0.05～0.2mgL
-1
4. the China fir clone according to claim 1 culture of rootage method that exsomatizes, it is characterized in that: in the step (2), the root induction medium is 1/2MS, and plant growth regulator is IBA0.05～0.2mgL
-1With NAA0.05～0.4mgL
-1Mixing.
5. the China fir clone according to claim 1 culture of rootage method that exsomatizes, it is characterized in that: in the step (2), the root induction medium is 1/4MS, and plant growth regulator is IBA0.05～0.2mgL
-1With NAA0.05～0.4mgL
-1Mixing.
6. the China fir clone according to claim 1 culture of rootage method that exsomatizes; It is characterized in that: in the step (3); In transplantation rooting test-tube plantlet the last week, rooting tube plantlet put under the natural lighting refine seedling, unclamp blake bottle day by day and seal film until removing at last.
7. the China fir clone according to claim 1 culture of rootage method that exsomatizes, it is characterized in that: in the step (3), the root length of test-tube plantlet is not less than 1cm.
8. according to claim 1 or the stripped culture of rootage method of 7 described China fir clones, it is characterized in that: in the step (3), the root length of test-tube plantlet is 1～3cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110129388A CN102217539B (en) | 2011-05-19 | 2011-05-19 | Isolated rooting culture method for fir clone |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110129388A CN102217539B (en) | 2011-05-19 | 2011-05-19 | Isolated rooting culture method for fir clone |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102217539A CN102217539A (en) | 2011-10-19 |
| CN102217539B true CN102217539B (en) | 2012-10-10 |
Family
ID=44774454
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110129388A Expired - Fee Related CN102217539B (en) | 2011-05-19 | 2011-05-19 | Isolated rooting culture method for fir clone |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102217539B (en) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657082B (en) * | 2012-04-28 | 2013-07-10 | 湖南农业大学 | In-vitro culture and planting regeneration and propagation method of Xianglei honeysuckle leaves and culture medium |
| CN102726299B (en) * | 2012-07-05 | 2013-08-07 | 福建农林大学 | Rapid two-step tissue culture propagation method of China fir based on LED (light emitting diode) light source |
| CN102960247B (en) * | 2012-11-23 | 2013-12-04 | 广西壮族自治区林业科学研究院 | Method for obtaining China fir tissue cultured explants |
| CN103109747B (en) * | 2013-03-10 | 2014-07-09 | 通化师范学院 | Rapid pseudolarix propagation method based on stem node propagation |
| CN103229723B (en) * | 2013-05-23 | 2014-07-30 | 广西壮族自治区林业科学研究院 | Rooting induction method of test-tube plantlet of cunninghamia lanceolata |
| CN103283598B (en) * | 2013-05-23 | 2015-04-29 | 广西壮族自治区林业科学研究院 | Primary fir wood culture bud induction method |
| CN104145822B (en) * | 2014-08-29 | 2016-03-23 | 福建省林业科学研究院 | The tissue cultivating and seedling method of a kind of excellent Chinese fir clone 11C |
| CN104304000B (en) * | 2014-08-29 | 2016-08-31 | 福建省林业科学研究院 | A kind of Clones of Cunninghamia Lanceolata tissue cultured seedling root induction method and root media |
| CN104255524A (en) * | 2014-10-15 | 2015-01-07 | 广西大学 | Method for quickly producing Chinese fir seedlings through micro-cutting |
| CN104429956A (en) * | 2014-11-24 | 2015-03-25 | 广西大学 | Method for test-tube rooting of cunninghamia lanceolata tissue culture seedlings by using compound LED light source |
| CN104686326A (en) * | 2015-02-13 | 2015-06-10 | 福建农林大学 | Method for promoting rooting of China fir tissue culture seedling |
| CN105145363B (en) * | 2015-09-16 | 2017-08-22 | 广西壮族自治区国有黄冕林场 | It is a kind of to significantly improve the method that China fir tissue culture produces emergence rate |
| CN106376461B (en) * | 2016-08-29 | 2019-03-05 | 广东省林业科学研究院 | The method of China fir tissue-cultured seedling culture of rootage and hardening |
| CN106922534A (en) * | 2017-03-27 | 2017-07-07 | 河池乐康生态农业科技有限公司 | A kind of method that tissue cultures quickly breed China fir |
| CN108040715A (en) * | 2017-12-04 | 2018-05-18 | 大新县科学技术情报研究所 | A kind of mountain planting method of buerretiodendron hsienmu |
| CN109479713A (en) * | 2018-12-03 | 2019-03-19 | 南京林业大学 | A kind of coppice spout using China fir carries out root induction method as explant |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101027972B (en) * | 2006-08-01 | 2011-04-20 | 厦门涌泉科技有限公司 | Culture medium propagation method of fir clone |
-
2011
- 2011-05-19 CN CN201110129388A patent/CN102217539B/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| 庞丽等.杉木优良无性系组培苗诱导根的研究.《江西农业大学学报》.2008,第30卷 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102217539A (en) | 2011-10-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102217539B (en) | Isolated rooting culture method for fir clone | |
| CN102150624B (en) | Tissue culture and rapid propagation method for pinellia tuber plant | |
| CN101584297B (en) | Tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana | |
| CN102948367B (en) | Method for performing in-vitro culturing and rapid propagating on bergenia crassifolia | |
| CN104255524A (en) | Method for quickly producing Chinese fir seedlings through micro-cutting | |
| CN104604677B (en) | A kind of tissue culture propagation method reducing russian dandelion tissue culture of Russia melting brown rate | |
| CN104221859B (en) | A kind of fast numerous cultural method of Garcinia mangostana | |
| CN102210267B (en) | Method for regenerating rose into complete plant | |
| CN103385168A (en) | Method for regeneration plant of tung oil tree leaf | |
| CN105052738A (en) | Tamarix chinensis tissue culture and rapid propagation method | |
| CN105265316A (en) | Rapid propagation method for plateaus of alliums | |
| CN105210870A (en) | The tissue culture propagation technology of No. 1, the anti-anvil of peach in Peach rootstock | |
| CN101589690A (en) | A kind of method of efficiently inducing Japanese red pine (Pinus densiflora) tissue cultivating seedling adventive root to take place | |
| CN105265317B (en) | A kind of For-carrying green onions rapid propagation method | |
| CN112715367B (en) | Method for carrying out Maozu secondary proliferation by using lanthanum nitrate | |
| CN106472306A (en) | One kind begins to flourish Herba Dendrobii high quality seedling asexual clonal method for quickly breeding | |
| CN104221864B (en) | A kind of cryptomeria clone isolated rooting culture method | |
| CN104115752B (en) | The tissue culture propagation of a kind of middle mountain China fir kind 118 | |
| CN103109728B (en) | Rapid seedling culturing method of pinus sylvestris in tube | |
| CN103155869A (en) | Sweet cherry rootstock Colt tissue culture method | |
| CN102613087A (en) | Method for culturing and breeding Correa carmen by using biological tissue | |
| CN106804426A (en) | Promote the box set and method of Companumoea root vitro proliferation | |
| CN104094848A (en) | Induction of tung tree hypocotyls callus and method for efficiently regenerating plants | |
| Cai et al. | Plant regeneration from cell suspension-derived protoplasts of Populus× beijingensis | |
| CN103828710B (en) | A kind of method of high efficiency chrysanthemum crossbreeding efficiency |
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 | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20111019 Assignee: Chongqing Jingwei Forestry Technology Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000370 Denomination of invention: Isolated rooting culture method for fir clone Granted publication date: 20121010 License type: Common License Record date: 20181211 Application publication date: 20111019 Assignee: Nanjing Baikang Biotechnology Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000369 Denomination of invention: Isolated rooting culture method for fir clone Granted publication date: 20121010 License type: Common License Record date: 20181211 |
|
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20111019 Assignee: Fujian Jinshuo Biotechnology Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000393 Denomination of invention: Isolated rooting culture method for fir clone Granted publication date: 20121010 License type: Common License Record date: 20181214 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EC01 | Cancellation of recordation of patent licensing contract |
Assignee: Fujian Jinshuo Biotechnology Co.,Ltd. Assignor: NANJING FORESTRY University Contract record no.: 2018320000393 Date of cancellation: 20211213 |
|
| EC01 | Cancellation of recordation of patent licensing contract | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20121010 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |