CN104054549B - The method of rich No. 6 tissue-culturing rapid propagations of a kind of apple variety cigarette - Google Patents
The method of rich No. 6 tissue-culturing rapid propagations of a kind of apple variety cigarette Download PDFInfo
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- CN104054549B CN104054549B CN201410333455.5A CN201410333455A CN104054549B CN 104054549 B CN104054549 B CN 104054549B CN 201410333455 A CN201410333455 A CN 201410333455A CN 104054549 B CN104054549 B CN 104054549B
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Abstract
The invention discloses the method for rich No. 6 tissue-culturing rapid propagations of a kind of apple variety cigarette, the method of rich No. 6 tissue-culturing rapid propagations of this apple variety cigarette is the medium Initial culture of MS+6-BA0.5mg/L+IBA0.1mg+PVP500mg/L, the squamous subculture adopting MS+6-BA0.5mg/L+IBA0.1mg/L process, the culture of rootage of employing root media 1/2MS+IBA1.0mg/L, the acclimatization and transplants with pure sawdust by adopting medium; Achieve rich No. 6 of apple variety cigarette to breed fast, not only ensure that higher survival rate, and remain the good characteristic of kind, for the development of Apple Industry provides method and access easily with expanding further.
Description
Technical field
The invention belongs to apple nursery plant cultivation technology field, particularly relate to the method for rich No. 6 tissue-culturing rapid propagations of a kind of apple variety cigarette.
Background technology
Apple Industry occupies very important status in Chinese agriculture economy, and Chinese apple cultivated area 225.3 ten thousand hectares in 2013, output is 3,170 ten thousand tons.Account for 1/2 of Gross World Product.At present, China apple production pattern by traditional poor efficiency closing orchard pattern progressively change the intensive High-efficient Production cultivation mode of modernization short anvil dense planting into and by apple production big country of the world progressively change into Apple Industry make the country prosperous.Apple nursery stock is the basis of apple production cultivation, and the quality of nursery stock directly affects the growth of apple tree, blooms, result and resistance, resistance against diseases and life-span.After nursery stock is subject to infecting of virus, tree body normal cell proliferation, will be damaged, and serious words can affect the normal physiological mechanism such as the growth potential of fruit tree, fruit quality and output, and this becomes the hidden danger of Chinese apple industry development.One period is the critical period that China's apple updates on a large scale from now on, and how providing support from nursery stock aspect and ensureing just becomes a problem being worth paying much attention to.No. 6, cigarette richness is the excellent bud mutation of Nagafu No.2.Compared to maternal plant Nagafu No.2, rich No. 6 of cigarette is Spur Type bud mutation, and early, early yield is high for treelet phase result, and fruit is individual large, well painted, obtains liking of numerous orchard workers.The present invention sets up the method for the rich No. 6 Fast-propagation tissue-culturing rapid propagations of a kind of apple variety cigarette, for production high-quality Spur-type Apples nursery stock, significant, wide market.
Summary of the invention
The object of the embodiment of the present invention is to provide the method for rich No. 6 tissue-culturing rapid propagations of a kind of apple variety cigarette, is intended to, for the cultivation of China's apple nursery stock provides a kind of method of tissue-culturing rapid propagation, update on a large scale to accelerate China's apple.
The embodiment of the present invention is achieved in that the method for rich No. 6 tissue-culturing rapid propagations of a kind of apple variety cigarette, and the method for rich No. 6 tissue-culturing rapid propagations of this apple variety cigarette comprises the following steps:
Step one, with No. 6, apple variety cigarette richness for material, the healthy and strong elite stand without damage by disease and insect of growth selection, 1 year raw branch is got March, carry out water planting in the greenhouse placing 25 DEG C, every premium on currency contains 30g sucrose, changes a water and cut old clip every 3d, when bud pumping on the annotinous branch of water planting is 1.5 ~ 2.0cm, clip tender shoots, removes and launches blade, under running water, rinse 6 ~ 8h, with 75% ethanol postincubation 8s, 0.1% mercuric chloride process 7min;
Step 2, after disinfection, 3min is cleaned with supersonic wave cleaning machine, use aseptic water washing again 3 ~ 4 times, be inoculated into by the tender shoots handled well in the medium of MS+6-BA0.5mg/L+IBA0.1mg+PVP (polyvinylpyrrolidone) 500mg/L and cultivate, often liter of medium adds 30g sucrose, 8g agar, then be placed on illumination 16h/d, illumination 2000lux, cultivate under 26 DEG C of conditions;
Step 3, by the cigarette rich No. 6 aseptic explants inoculation MS+6-BA0.5mg/L+IBA0.1mg/L agar 8g/L obtained, carries out squamous subculture in the medium of illumination 2000lux, often liter of additional 30g sucrose, 8g agar;
Step 4, select the robust growth that squamous subculture obtains, be highly greater than 1.5cm, growth time is bud of 20 days, is inoculated in 1/2MS+IBA1.0mg/L (sucrose 30g/L, agar 8g/L) root media and carries out culture of rootage;
Step 5, after plantlet in vitro root length reaches 2cm, move on to outdoor to shelter from heat or light hardening about 10 ~ 12 days, then opened by culture vessel bottleneck, carry out uncork hardening after 2 ~ 4 days under natural daylight, seedling of taking root takes out from medium, putting into concentration is that 0.1% carbendazim solution soaks 5min, and medium unnecessary for root is cleaned up, finally rinse 2 ~ 3 times with clear water, in the nutritive cube be then transplanted to pure sawdust.
Further, in step 5, the way to manage of transplanted seedling is: seedling of taking root after transplanting manages under shade net, within first week, adopts concentration to be that 0.1% carbendazim solution is sprayed, and the sprinkling irrigation of second week use every day water once, keeps transplanting medium moistening ventilative.Offspring pumping to be generated is extensible shade net after going out sprouting.
The method of rich No. 6 tissue-culturing rapid propagations of apple variety cigarette provided by the invention, be the medium Initial culture of MS+6-BA0.5mg/L+IBA0.1mg+PVP500mg/L by employing medium, survival rate is 30.77%, is conducive to the foundation of rich No. 6 plantlet in vitro of cigarette; Adopt the squamous subculture of MS+6-BA0.5mg/L+IBA0.1mg/L process, expand numerous coefficient up to 5.64; Adopt the culture of rootage of root media 1/2MS+IBA1.0mg/L and rooting rate reaches 45.8%; Acclimatization and transplants is carried out, survival rate 75% with pure sawdust; That repeats carries out squamous subculture to rich No. 6 kinds of cigarette, and culture of rootage and transplanting can realize the breeding fast of the rich No. 6 virus-free nursery stocks of apple variety cigarette in the short period of time.Remained the good characteristic of rich No. 6 kinds of cigarette by tissue culture technique, be conducive to the preservation of rich No. 6 improved seeds of cigarette, the virus-free nursery stock for rich No. 6 of apple cigarette provides efficient raising technology easily, promotes further developing of Apple Industry.
Accompanying drawing explanation
Fig. 1 is the method flow diagram of rich No. 6 tissue-culturing rapid propagations of apple variety cigarette that the embodiment of the present invention provides.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Below in conjunction with drawings and the specific embodiments, application principle of the present invention is further described.
As shown in Figure 1, the method for rich No. 6 tissue-culturing rapid propagations of the apple variety cigarette of the embodiment of the present invention comprises the following steps:
S101: with No. 6, apple variety cigarette richness for material, the healthy and strong elite stand without damage by disease and insect of growth selection, gets 1 year raw branch March, carries out water planting in the greenhouse placing 25 DEG C, every premium on currency contains 30g sucrose, change a water every 3d and cut old clip, when the bud pumping on the annotinous branch of water planting is 1.5 ~ 2.0cm, clip tender shoots, remove and launch blade, 6 ~ 8h is rinsed under running water, with 75% ethanol postincubation 8s, 0.1% mercuric chloride process 7min;
S102: after disinfection, 3min is cleaned with supersonic wave cleaning machine, use aseptic water washing again 3 ~ 4 times, the tender shoots handled well is inoculated in the medium of MS+6-BA0.5mg/L+IBA0.1mg+PVP (polyvinylpyrrolidone) 500mg/L and cultivates, often liter of medium adds 30g sucrose, 8g agar, is then placed on illumination 16h/d, illumination 2000lux, cultivates under 26 DEG C of conditions;
S103: by the cigarette rich No. 6 aseptic explants inoculation best medium process obtained, i.e. MS+6-BA0.5mg/L+IBA0.1mg/L agar 8g/L, carries out squamous subculture in the medium of illumination 2000lux, often liter of additional 30g sucrose, 8g agar;
S104: select the robust growth that squamous subculture obtains, be highly greater than 1.5cm, growth time is bud of 20 days, is inoculated in 1/2MS+IBA1.0mg/L (sucrose 30g/L, agar 8g/L) root media and carries out culture of rootage;
S105: after plantlet in vitro root length reaches 2cm, move on to outdoor to shelter from heat or light hardening about 10 ~ 12 days, then culture vessel bottleneck is opened, uncork hardening is carried out after 2 ~ 4 days under natural daylight, seedling of taking root takes out from medium, and putting into concentration is that 0.1% carbendazim solution soaks 5min, and medium unnecessary for root is cleaned up, finally rinse 2 ~ 3 times with clear water, in the nutritive cube be then transplanted to pure sawdust.
In step S105, the way to manage of transplanted seedling is: seedling of taking root after transplanting manages under shade net, within first week, adopts concentration to be that 0.1% carbendazim solution is sprayed, and the sprinkling irrigation of second week use every day water once, keeps cultivation matrix moistening ventilative.Offspring pumping to be generated is extensible shade net after going out sprouting.
Concrete steps of the present invention are as follows:
Step one, Initial culture
Rich No. 6 of apple variety cigarette is material, the healthy and strong elite stand without damage by disease and insect of growth selection, 1 year raw branch is got March, water planting (every premium on currency contains 30g sucrose) is carried out in the greenhouse placing 25 DEG C, change a water every 3d and cut old clip, when bud pumping on the annotinous branch of water planting is 1.5 ~ 2.0cm, clip tender shoots, remove and launch blade, 6 ~ 8h is rinsed under running water, with 75% ethanol postincubation 8s, 0.1% mercuric chloride process 7min, after disinfection, 3min is cleaned with supersonic wave cleaning machine, use aseptic water washing again 3 ~ 4 times, the tender shoots handled well is inoculated into MS+6-BA0.5mg/L+IBA0.1mg/L+ variable concentrations process (0, 0.3, 0.5, 0.7, cultivate in the medium of polyvinylpyrrolidone (PVP) 1.0g/L), often liter of medium adds 30g sucrose, 8g agar, each triangle bottle graft 3 buds, often process 15 bottles, then illumination 16h/d is placed on, illumination 2000lux, cultivate under 26 DEG C of conditions, pollution rate is investigated respectively after 14d, melting brown rate, survival rate, the Initial culture of rich No. 6 of smoke be optimum medium is MS+6-BA0.5mg/L+IBA0.1mg+PVP500mg/L (sucrose 30g/L, agar 8g/L),
Step 2, squamous subculture
The aseptic explant of acquisition is inoculated into containing the capable squamous subculture of variable concentrations 6-BA, often liter of additional 30g sucrose, 8g agar, every bottle of 3 strains, each process 10 bottles, repeats 3 times, and the best medium that cigarette is rich No. 6 is, i.e. MS+6-BA0.5mg/L+IBA0.1mg/L (agar 8g/L, illumination 2000lux);
Step 3, culture of rootage
Select the robust growth that squamous subculture obtains, highly be greater than the plant of 1.5cm, growth time is bud of 20 days, be inoculated in 1/2MS+ variable concentrations IBA process (0,1.0,0.9,1.0,1.1mg/L) culture of rootage in carry out culture of rootage, every bottle graft kind 3, each process 10 bottles, repeats 3 times, obtaining the rich No. 6 optimum root medias of cigarette is 1/2MS+IBA1.0mg/L (sucrose 30g/L, agar 8g/L);
Step 4, acclimatization and transplants
After plantlet in vitro root length reaches 2cm, move on to outdoor to shelter from heat or light hardening about 10 ~ 12 days, then culture vessel bottleneck is opened, uncork hardening is carried out after 2 ~ 4 days under natural daylight, seedling of taking root takes out from medium, putting into concentration is that 0.1% carbendazim solution soaks 5min, and medium unnecessary for root is cleaned up, finally rinse 2 ~ 3 times with clear water, then different substrates process (the pure sawdust that the capacity that is transplanted to is identical, sawdust+matrix, pure matrix, matrix+perlite) nutritive cube in, the way to manage of transplanted seedling is: seedling of taking root manages under shade net, within first week, concentration is adopted to be that 0.1% carbendazim solution is sprayed, the sprinkling irrigation of second week use every day water once, keep transplanting medium moistening ventilative.Added up acclimatization and transplants survival rate afterwards in 14 days, draw by the acclimatization and transplants survival rate of pure sawdust best.
By following experiment and data, principle of the present invention is described further:
1, reference table 1 variable concentrations PVP process is on the impact of explant growth;
Table 1 variable concentrations PVP process is on the impact of explant growth
Note: represent difference not significantly (P>0.05) with columns same letter, different letter representation significant difference (P<0.05).Data are Mean ± SE (mean value ± standard error); Lower same.
2, the process of different 6-BA concentration is on the impact of rich No. 6 squamous subculture of cigarette:
The different 6-BA concentration of table 2 is on the impact of rich No. 6 squamous subculture of cigarette
| Minimal medium | 6-BA(mg/L) | IBA(mg/L) | Expand numerous coefficient |
| MS | 0 | 0.1 | 1.25±0.06c |
| MS | 0.3 | 0.1 | 1.41±0.09c |
| MS | 0.4 | 0.1 | 2.78±0.11b |
| MS | 0.5 | 0.1 | 5.64±0.22a |
| MS | 0.6 | 0.1 | 4.32±0.14a |
3, the process of different I BA concentration is on rich No. 6 impacts of taking root of cigarette:
The process of table 3 different I BA concentration is on rich No. 6 impacts of taking root of cigarette
4 different hardening matrix are on the impact of seedling growth of taking root:
The impact that the different cultivation matrix process of table 4 grows transplanted seedling
| Matrix treatments | Rich No. 6 transplanting survival rates of cigarette |
| Sawdust | 75.00±4.23a |
| Sawdust+matrix | 67.50±2.42b |
| Matrix | 55.00±1.32c |
| Matrix+perlite | 57.50±5.32c |
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (1)
1. a method for rich No. 6 tissue-culturing rapid propagations of apple variety cigarette, is characterized in that, the method for rich No. 6 tissue-culturing rapid propagations of this apple variety cigarette comprises the following steps:
Step one, with No. 6, apple variety cigarette richness for material, cleans 3min with supersonic wave cleaning machine, then uses aseptic water washing 3 ~ 4 times, be inoculated into by the tender shoots handled well in the medium of polyvinylpyrrolidone 500mg/L and cultivate; Often liter of medium adds 30g sucrose, 8g agar, is then placed on illumination 16h/d, illumination 2000lux, cultivates under 26 DEG C of conditions;
Step 2, is inoculated in the medium of MS+6-BA0.5mg/L+IBA0.1mg/L by the aseptic explant of acquisition, carries out squamous subculture, adds 30g sucrose in often liter of medium, 8g agar under illumination 2000lux condition;
Step 3, select the robust growth that squamous subculture obtains, be highly greater than the plant of 1.5cm, growth time is bud of 20 days, is inoculated in 1/2MS+IBA1.0mg/L root media and carries out culture of rootage, often liter of medium supplemented 30g sucrose, 8g agar;
Before step one is with supersonic wave cleaning machine cleaning, with No. 6, apple variety cigarette richness for material, the healthy and strong elite stand without damage by disease and insect of growth selection, in clip in March 1 year raw branch, water planting is carried out in the greenhouse placing 25 DEG C, water planting is that every premium on currency contains 30g sucrose, change a water every 3d and cut the old clip of water planting branch, when bud pumping on the annotinous branch of water planting is 1.5 ~ 2.0cm, clip tender shoots, removes and launches blade, under running water, rinse 6 ~ 8h, with 75% ethanol postincubation 8s, 0.1% mercuric chloride process 7min;
After the culture of rootage step of step 3, plantlet in vitro root length reaches 2cm, move on to outdoor to shelter from heat or light hardening 10 ~ 12 days, then opened by culture vessel bottleneck, carry out uncork hardening after 2 ~ 4 days under natural daylight, seedling of taking root takes out from medium, putting into concentration is that 0.1% carbendazim solution soaks 5min, and medium unnecessary for root is cleaned up, finally rinse 2 ~ 3 times with clear water, in the nutritive cube be then transplanted to pure sawdust;
The way to manage of transplanted seedling is: seedling of taking root after transplanting manages under shade net, within first week, concentration is adopted to be that 0.1% carbendazim solution is sprayed, once, keep transplanting medium moistening ventilative, namely offspring pumping to be generated shifts out shade net after going out sprouting to the sprinkling irrigation of second week use every day water.
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| CN106937597B (en) * | 2017-04-12 | 2018-11-27 | 山东农业大学 | A kind of tissue-culturing rapid propagation method suitable for high flavonoids apple new lines |
| CN109863932A (en) * | 2018-10-28 | 2019-06-11 | 陕西海棠生态农林股份有限公司 | A kind of method of nutritive cube cultivation Tissue-cultured apple seedling |
| CN109679993B (en) * | 2019-02-22 | 2020-11-27 | 北京林业大学 | Construction method of agrobacterium rhizogenes-mediated transgenic plant |
| CN110651711B (en) * | 2019-09-18 | 2022-05-13 | 西北农林科技大学 | Rapid propagation method for new variety of apples Ruiyang and Ruiyue |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4038778A (en) * | 1976-09-27 | 1977-08-02 | Gte Laboratories Incorporated | Tissue culture technique for asexual plant reproduction and a medium for use therewith |
| CN1443444A (en) * | 2003-04-28 | 2003-09-24 | 陕西师范大学 | Australian greening (Grany Smith) quick propagation method |
| CN103011969A (en) * | 2012-12-25 | 2013-04-03 | 浙江农林大学 | Medium specialized for plant tissue culture, and preparation method thereof |
| CN103798136A (en) * | 2013-12-11 | 2014-05-21 | 柳州赛特生物科技研发中心 | Special phalaenopsis tissue culture medium |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4038778A (en) * | 1976-09-27 | 1977-08-02 | Gte Laboratories Incorporated | Tissue culture technique for asexual plant reproduction and a medium for use therewith |
| CN1443444A (en) * | 2003-04-28 | 2003-09-24 | 陕西师范大学 | Australian greening (Grany Smith) quick propagation method |
| CN103011969A (en) * | 2012-12-25 | 2013-04-03 | 浙江农林大学 | Medium specialized for plant tissue culture, and preparation method thereof |
| CN103798136A (en) * | 2013-12-11 | 2014-05-21 | 柳州赛特生物科技研发中心 | Special phalaenopsis tissue culture medium |
Non-Patent Citations (1)
| Title |
|---|
| 苹果砧木组织培养与快繁技术研究;赵亮明等;《西北农业学报》;20111231;第20卷(第07期);118-122 * |
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