CN103250646B - Method for controlling rapid propagation of platycodon grandiflorum by virtue of light source - Google Patents
Method for controlling rapid propagation of platycodon grandiflorum by virtue of light source Download PDFInfo
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- CN103250646B CN103250646B CN201310206548.7A CN201310206548A CN103250646B CN 103250646 B CN103250646 B CN 103250646B CN 201310206548 A CN201310206548 A CN 201310206548A CN 103250646 B CN103250646 B CN 103250646B
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Abstract
The invention discloses a method for controlling the rapid propagation of platycodon grandiflorum by virtue of a light source. The method comprises the following steps of: (1) selecting seeds of the platycodon grandiflorum, cleaning, disinfecting, inoculating the seeds onto an MS culture medium to obtain tissue culture seedlings, grafting stems on the tissue culture seedlings onto a new MS culture medium, culturing the grafted tissue culture seedlings in a dark environment, then transferring the new MS culture medium with the seedlings in the dark environment under a white fluorescent lamp for culturing to obtain the stems which have the lengths of 10-15mm and respectively have two euphyllas; and (2) selecting the stems obtained in the step (1) and inoculating the selected stems onto a culture medium with 1/2MS+0.1mg/l of NAA (neutron activation analysis), culturing in the dark environment for 5-7 days, transferring the culture medium with the seedlings under the light-emitting diode (LED) light source and culturing for 35-45 days to obtain tissue culture seedling plants of the platycodon grandiflorum, wherein the LED light source sends combined light of one or two of red light or blue light with different proportions, the light intensity is 45-55mu mol.m<-2>.S<-1>, the photoperiod is 8-12 hours, and the temperature is 25+/-2 DEG C. The method is easy to operate, enables the plants to rapidly grow in a healthy and strong way, and environment-friendly, the saving is realized.
Description
Technical field
The present invention relates to and a kind ofly carry out balloonflower root method numerous soon by means of light source control, belong to biological technical field.
Background technology
The light source used in Plant Tissue Breeding is generally fluorescent lamp, but power consumption is large, and emission spectrum can not match with the photosynthesis absorption spectrum of plant well, and containing much unnecessary wavelength, light efficiency is very low; And produce a lot of thermal radiation, and can not to plant close to irradiating, not high to plant growth light stimulus efficiency, service life is not long yet.Compare with traditional light source, light emitting diode (LED) receives publicity day by day as a kind of novel semi-conductor light source that can be used for plant irradiation in recent years.The basic structure of LED is one block of electroluminescent semi-conducting material, is placed on a leaded shelf, surrounding epoxy sealing, plays the effect of protection internal core.LED ripple width, photochromic pure, wavelength abundant species, coincide with the spectral range of photosynthesis of plant and morphogenesis.LED also has less quality and volume, longer life-span, regulates its frequency and dutycycle can save the energy and improve photosynthetic efficiency.Due to these significant features, LED is subject to increasing favor in recent years in Plant Tissue Breeding.
LED is used for Plant Tissue Breeding light source and has following advantage: LED light exports half width, close to monochromatic light, be used singly or in combination, use LED can concentrate the light balancedly irradiate crops of specific wavelength, not only can regulate and control the physiological activities such as C N metabolism thus regulate the morphological indexs such as plant height dry weight and fresh weight of plant seedlings, plant leaf stomatal aperture can also being regulated, the physiological activities such as improving activity of root system, preparing for obtaining the transplanting of healthy and strong plantlet in vitro.The LED applied in plant tissue culture in addition belongs to cold light source, and illumination system heating is few, achieves low heat loads, can be placed in from plant very close to place and can not roasting for plant wound; LED shape is minimum, the LED combination of different wave length can be prepared into the device of multiple ratio various shape, take up room little, easy for installation, conveniently filters out the specific light source of applicable certain species plant; In addition, its stronger durability also reduces operation cost.
Balloonflower root (Platycodon grandiflorum) belongs to Campanulaceae, herbaceos perennial, adularescent milk.Root is conical, and stem is upright, and base portion impeller is raw or to life.The effect that there is antibechic and eliminate the phlegm.Its bud, flower shape are unique, and color is gorgeous, has much sight; Root contains multiple saponin(e, tool a surname lung, loose to tremble with fear, eliminate the phlegm, the effect such as apocenosis; Root and tender leaf also can processing and eating.Balloonflower root is collection reward, food, medicine are multipurpose economic plants all over the body, has great value of exploiting and utilizing.Generally commonly use seminal propagation or bury root propagation, but in seminal propagation process, the phenomenon such as quality deterioration, variation can be produced, and raw seedling growing way is weak then, cannot form higher output, long-term bury root nourish and generate in process, virus infections can be subject to, production declining.
This method explores the screening of balloonflower root tissue culture sprout quick propagation light source, establishes complete feasible test method, and operating procedure is simple, and test effect is stable, clear, therefore no matter can well be applied in scientific research, detection and student's test.
Summary of the invention
For solving the problem, we send out strong sprout aseptic for material with balloonflower root kind, carry out light quality process, obtain the optimum spectral power distribution of the most applicable balloonflower root plantlet in vitro physiological growth, to providing technical support for the Germ-plasma resources protection of balloonflower root, new varieties and popularization and factorial praluction.The object of the present invention is to provide the one more effective balloonflower root tissue culture sprout quick propagating technology by means of light source control.
Carry out a balloonflower root technology numerous soon by means of light source control, comprise the following steps:
(1) preparation of explant: choose Seed of Platycodon Grandiflorum, cleaning, sterilization, seed is inoculated into cultivation on MS medium and obtains plantlet in vitro, stem section on plantlet in vitro is transferred on new MS medium, first cultivate under dark, then cultivate under proceeding to white fluorescent lamp, obtain 10 ~ 15mm long, with the stem section of two panels true leaf;
(2) light quality process: the stem section that selecting step (1) obtains is seeded on the medium of 1/2MS+0.1mg/l NAA, first cultivate 4 ~ 7 days under dark, then cultivate 35 ~ 45 days under proceeding to LED light source, obtain healthy and strong balloonflower root plantlet in vitro plant, wherein said LED light source is the combined light of one or both different proportions of ruddiness or blue light, condition of culture: light intensity 45 ~ 55 μm of olm
-2s
-1, the photoperiod is 8 ~ 12 hours, temperature 25 ± 2 DEG C.
Described step (1) cleaning and sterilizing process is specific as follows: Seed of Platycodon Grandiflorum first rinses 2 ~ 3 hours with water, then in superclean bench, use the alcohol disinfecting 20 ~ 30 seconds of 75% successively, the hypochlorite disinfectant of 15% 10 ~ 15 minutes, the hydrogen peroxide sterilization of 15% 10 ~ 15 minutes, finally uses aseptic water washing 5 ~ 6 times.
Described in step (1), stem section is cultivated 4 ~ 7 days under dark; The condition of culture that described stem section is cultivated under white fluorescent lamp: light intensity 45 ~ 55 μm of olm
-2s
-1, the photoperiod is 8 ~ 12 hours, temperature 25 ± 2 DEG C.
Described LED light source be ultra-blue-light (B), monochromatic ruddiness (R), Lan Hong than 5:3 (blue light and ruddiness portfolio ratio 5:3/BR53), Lan Hong than 1:1 (blue light and ruddiness portfolio ratio 1:1/BR11) or blue red in 3:5 (blue light and ruddiness portfolio ratio 3:5/BR35), preferably bluely redly compares 5:3.
Described LED light source is the rectangle lamp plate of a 47.5 × 50cm, the length direction of described rectangle lamp plate arranges 20 Luminous cover groups, the width of described rectangle lamp plate arranges 19 Luminous cover groups, and each Luminous cover group is made up of three light emitting diodes.
Described LED light source controls by an adjustable constant-flow driving power, regulates and controls light intensity by regulate electrical current.
Beneficial effect of the present invention:
The invention provides and a kind ofly carry out balloonflower root technology numerous soon by means of light source control.The inventive method has the following advantages: 1) simple to operate, effective; 2) energy environment protection is saved; 3) plant strain growth is healthy and strong fast.The inventive method and result are carry out relevant biological study numerous with balloonflower root tissue culture sprout quick and develop to provide new approach and foundation.
Accompanying drawing explanation
Fig. 1, Different Light is on the impact of the whole strain fresh weight of balloonflower root plantlet in vitro.
Fig. 2, Different Light is on the impact of the whole strain dry weight of balloonflower root plantlet in vitro.
Fig. 3, Different Light falls the impact of three leaf leaf dry weights on balloonflower root plantlet in vitro.
Fig. 4, Different Light falls the impact of three leaf leaf areas on balloonflower root plantlet in vitro.
Fig. 5, the impact that Different Light is long on balloonflower root plantlet in vitro stem.
Fig. 6, different light source process is on the impact of balloonflower root plantlet in vitro VC content.
Fig. 7, different light source process is on the impact of balloonflower root plantlet in vitro improving activity of root system.
Fig. 8, different light source process is on the impact of balloonflower root morphology of stomata.In Fig. 8, Sto represents pore, and Ep represents epidermal cell.
Embodiment
In following embodiment, method therefor is conventional method if no special instructions, and described percentage composition is volumn concentration if no special instructions.
1, first use the inventive method, by means of LED light source, fast numerous process carried out to balloonflower root (production kind), comprise following two steps:
(1) preparation of explant: choose the Seed of Platycodon Grandiflorum that clean full color and luster is brighter, first rinse 2 hours with water, then in superclean bench with 75% alcohol disinfecting 30 seconds, the hypochlorite disinfectant of 15% 15 minutes, the hydrogen peroxide of 15% is sterilized 15 minutes, aseptic water washing 5 times, seed is inoculated on MS medium; After 30 days, the stem section on plantlet in vitro is transferred on new MS medium, and dark lower cultivation, after 5 days, is cultivated under proceeding to white fluorescent lamp, condition of culture: light intensity 50 μm of olm
-2s
- 1, the photoperiod is 12 hours, temperature 25 ± 2 DEG C, obtain 15mm long, with the stem section of two panels true leaf;
(2) light quality process: the stem section that selecting step (1) obtains is seeded on the medium of 1/2MS+0.1mg/l NAA, first cultivate 5 days under dark, then proceed to ultra-blue-light (B), Lan Hong cultivates 40 days than under the LED light source of 3:5 (BR35), monochromatic ruddiness (R) than 5:5 (BR55), Lan Hong than 5:3 (BR53), Lan Hong, white fluorescent lamp (FL) in contrast, condition of culture: light intensity 50 μm of olm
-2s
-1, the photoperiod is 12 hours, temperature 25 ± 2 DEG C, obtains balloonflower root plantlet in vitro plant.
2, light source: described LED light source is the rectangle lamp plate of a 47.5 × 50cm, the length direction of described rectangular plate lamp plate arranges 20 Luminous cover groups, at described rectangular plate lamp plate width, 19 Luminous cover groups are set, each Luminous cover group is made up of three light emitting diodes, the composition of lamp can be regulated according to different light source requirements, light intensity, light quality ratio.
Described LED light source controls by an adjustable constant-flow driving power, regulates and controls light intensity by regulate electrical current.
3, treatment effect
3.1 Different Light process are on the impact of balloonflower root plantlet in vitro form
See Fig. 1, Fig. 2, compared with Lan Hong to contrast with other LED light source process and white fluorescent lamp than the whole strain fresh weight of the lower balloonflower root plantlet in vitro of 5:3LED process and dry weight, without significant difference.See Fig. 3, the three leaf leaf dry weights that fall under blue-ray LED process are significantly higher than other process, and Lan Hong takes second place than 5:3, with other LED light source process without significant difference.See Fig. 4, three leaf leaf areas of falling under blue-ray LED process are maximum, and Lan Hong takes second place than 5:3LED, are all significantly higher than other LED light source process and white fluorescent lamp contrast.See Fig. 5, in all process, plant height is without significant difference, but blue red higher than the lower balloonflower root plantlet in vitro plant height of 1:1LED process than 5:3LED and Lan Hong.Result illustrates, Lan Hong is comparatively quicker and healthy and strong than the lower plant strain growth of 5:3LED process.
3.2 Different Light process are on the impact of balloonflower root plantlet in vitro VC content
See Fig. 6, balloonflower root plantlet in vitro VC content is at Lan Hong than the highest under 5:3LED, and white fluorescent lamp contrast is lower minimum, other value that mediates.Therefore, Lan Hong is conducive to improving the VC content in balloonflower root plantlet in vitro than 5:3LED.
3.3 Different Light process are on the impact of balloonflower root plantlet in vitro improving activity of root system
See Fig. 7, balloonflower root plantlet in vitro improving activity of root system, takes second place under blue-ray LED than maximum under 5:3LED at Lan Hong, is significantly higher than other LED process and white fluorescent lamp contrast.Therefore, Lan Hong is conducive to than 5:3LED the improving activity of root system improving balloonflower root plantlet in vitro.
3.4 Different Light process are on the impact of balloonflower root plantlet in vitro stomatal properties
See Fig. 8 and table 3, balloonflower root plantlet in vitro pore frequency, reduces than under 3:5LED than 5:3LED, blue-ray LED, Lan Hong at red-light LED, Lan Hong than maximum under 1:1LED successively at Lan Hong, minimum under white fluorescent lamp contrast.Stomatal size, reduces under blue-ray LED, Lan Hong are than 5:3LED, red-light LED than maximum under 3:5LED successively at Lan Hong, at Lan Hong than minimum under 1:1LED.Pore aspect ratio is maximum under Lan Hong is than 1:1LED and red-light LED, pore is in abnormal oblateness, pore aspect ratio at Lan Hong than minimum under 3:5LED, pore is in abnormal subcircular, and white fluorescent lamp contrast, blue-ray LED, Lan Hong to mediate value than 5:3LED air holes aspect ratio, in the ellipse with normal function.These results suggest that, Lan Hong is conducive to the normal development of balloonflower root plantlet in vitro pore than 5:3LED.
3.5 Different Light process are on the impact of balloonflower root plantlet in vitro C N metabolism
In table 1, under red-light LED, soluble sugar content is the highest, is significantly higher than other LED process and white fluorescent lamp contrast, without significant difference between other LED light source process.All process Determination of Free Amino Acids are without significant difference, and blue-ray LED, Lan Hong are than slightly high under 5:3LED.Soluble protein content is at blue-ray LED, Lan Hong than the highest under 1:1LED, and be significantly higher than other process, Lan Hong is than lower slightly under 5:3.In sum, Lan Hong is higher than C N metabolism level under 5:3LED.
3.6 Different Light process are on the impact of balloonflower root plantlet in vitro natomical leaf structure
In table 2, under blue-ray LED, balloonflower root plantlet in vitro vane thickness is maximum, and Lan Hong takes second place than under 1:1LED than 5:3LED and Lan Hong, and Lan Hong is than minimum under 3:5LED, and other LED light source process mediates value.Palisade tissue length is maximum under blue-ray LED, and white fluorescent lamp contrasts and indigo plant is red takes second place than under 5:3LED, and Lan Hong is than minimum under 3:5LED.Lan Hong, than having maximum spongy tissue thickness under 5:3LED, takes second place under blue-ray LED, is significantly higher than other LED process and white fluorescent lamp contrast.In the blade of many plants, palisade cell contains about 70% of chloroplast sum, and exceedes sponge cell about to twice.Palisade tissue impels light to focus on chloroplast better, and spongy tissue improves light capture ability by scattered light.So larger palisade tissue and spongy tissue thickness ensure that higher photosynthetic capacity.Therefore blue-ray LED, Lan Hong grow better than balloonflower root plantlet in vitro natomical leaf structure under 5:3LED.
The effect of 3.7 Different Light process balloonflower root plantlet in vitro
Lan Hong is than under 5:3LED process, and the blade of balloonflower root plantlet in vitro is large and thick, physically well develops, and photosynthetic capacity is strong, and leaf dry weight, whole strain dry weight and fresh weight of plant seedlings are comparatively large, and plant strain growth is healthy and strong; Soluble sugar, free amino acid, soluble protein content are higher, and improving activity of root system is the highest, and VC content is the highest.In a word, Lan Hong is than under 5:3LED process, and the growth of balloonflower root plantlet in vitro is quick, healthy and strong, and every physiological and biochemical index is substantially all higher than the process of contrast white fluorescent lamp, therefore, and the alternative source of light that Lan Hong can produce as balloonflower root plantlet in vitro than 5:3LED.Red and blue LED combined light source, can the advantage of comprehensive Red and blue light, makes up monochromatic shortcoming, the preferred light source can cultivated as balloonflower root plantlet in vitro.
Table 1. Different Light is on the impact of balloonflower root plantlet in vitro C N metabolism
Note: between the subscript representative process of different letter, there were significant differences, and a is maximum, and b takes second place, the like.
Table 2. Different Light is on the impact of balloonflower root plantlet in vitro natomical leaf structure
Table 3. Different Light is on the impact of balloonflower root plantlet in vitro pore
Claims (4)
1. carry out a balloonflower root method numerous soon by means of light source control, it is characterized in that comprising the following steps:
(1) preparation of explant: choose Seed of Platycodon Grandiflorum, cleaning, sterilization, seed is inoculated into cultivation on MS medium and obtains plantlet in vitro, stem section on plantlet in vitro is transferred on new MS medium, first cultivate 4 ~ 7 days under dark, then cultivate under proceeding to white fluorescent lamp, condition of culture: light intensity 45 ~ 55 μm of olm
-2s
-1, the photoperiod is 8 ~ 12 hours, temperature 25 ± 2 DEG C, obtain 10 ~ 15mm long, with the stem section of two panels true leaf;
(2) light quality process: the stem section that selecting step (1) obtains is seeded on the medium of 1/2MS+0.1mg/l NAA, first cultivate 4 ~ 7 days under dark, then cultivate 35 ~ 45 days under proceeding to LED light source, obtain healthy and strong balloonflower root plantlet in vitro plant, wherein said LED light source is blue red in 5:3, condition of culture: light intensity 45 ~ 55 μm of olm
-2s
-1, the photoperiod is 8 ~ 12 hours, temperature 25 ± 2 DEG C.
2. according to claim 1ly carry out balloonflower root method numerous soon by means of light source control, it is characterized in that described step (1) cleaning and sterilizing process is specific as follows: Seed of Platycodon Grandiflorum first rinses 2 ~ 3 hours with water, then in superclean bench, use the alcohol disinfecting 20 ~ 30 seconds of 75% successively, the hypochlorite disinfectant of 15% 10 ~ 15 minutes, the hydrogen peroxide sterilization of 15% 10 ~ 15 minutes, finally uses aseptic water washing 5 ~ 6 times.
3. according to claim 1ly carry out balloonflower root method numerous soon by means of light source control, it is characterized in that described LED light source is the rectangle lamp plate of a 47.5 × 50cm, the length direction of described rectangle lamp plate arranges 20 Luminous cover groups, the width of described rectangle lamp plate arranges 19 Luminous cover groups, and each Luminous cover group is made up of three light emitting diodes.
4. according to claim 1 or 3, carry out balloonflower root method numerous soon by means of light source control, described LED light source controls by an adjustable constant-flow driving power, regulates and controls light intensity by regulate electrical current.
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| CN107624648B (en) * | 2017-10-30 | 2020-05-01 | 宁夏西北药材科技有限公司 | Method for promoting rapid propagation of astragalus membranaceus by using LED lamp |
| CN114403008A (en) * | 2022-01-28 | 2022-04-29 | 中国科学院近代物理研究所 | Rapid platycodon grandiflorum radiation mutation breeding method |
| CN115918529B (en) * | 2022-09-28 | 2024-05-28 | 山东惠美农牧发展有限公司 | Lily test tube bulb proliferation method |
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Non-Patent Citations (3)
| Title |
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| Effects of different spectral lights on Oncidium PLBs induction,proliferation, and plant regeneration;Liu Mengxi等;《Plant Cell Tiss Organ Cult》;20111231;第106卷;第1-10页 * |
| Effects of Light Emitting Diodes on Growth and Morphogenesis of in vitro Seedlings in Platycodon grandiflorum;EUN Jong Seon等;《Korean J. Plant Tissue Culture》;20001231;第27卷(第1期);第71-75页 * |
| 桔梗的组织培养及快速繁殖;张凤生等;《北方园艺》;20071231(第1期);第165页 * |
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