CN103004589A - Tissue culture and rapid propagation method of agrimony - Google Patents
Tissue culture and rapid propagation method of agrimony Download PDFInfo
- Publication number
- CN103004589A CN103004589A CN2012105058692A CN201210505869A CN103004589A CN 103004589 A CN103004589 A CN 103004589A CN 2012105058692 A CN2012105058692 A CN 2012105058692A CN 201210505869 A CN201210505869 A CN 201210505869A CN 103004589 A CN103004589 A CN 103004589A
- Authority
- CN
- China
- Prior art keywords
- agrimony
- tissue
- cultivated
- medium
- plant growth
- 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.)
- Granted
Links
Abstract
The invention relates to a tissue culture and rapid propagation method of plants and particularly relates to a tissue culture and rapid propagation method of tulipa edulis. The method comprises the following steps that firstly, explants are disinfected, wherein the large leaflets of pinnately compound leaves of the fully-developed tulipa edulis are washed and disinfected and then are cut into small pieces of 1 to 3cm<2>, and then the small pieces with upward front surfaces are horizontally inoculated into an MS (Murashige and Skoog) culture medium with different plant growth regulating substances; secondly, adventitious bud is induced, wherein the explants are inoculated into the MS culture medium; and finally, the adventitious buds of the leaves, which are light green, grow vigorously and are 2cm to 3cm high, are cut, the adventitious buds are inserted into a rooting medium, the adventitious buds are taken out from the medium after 20-30 hours, washing the medium adhered to the roots of the adventitious buds are washed, the adventitious buds are transplanted into earth, and a beaker is removed after young leaves grow. The tissue culture and rapid propagation method has the advantages that the rooting rate and the rotting number are increased, the length and thickness of roots are moderate, and the survival rate of transplanting is greatly increased.
Description
Technical field
The present invention relates to Plant Tissue Breeding and method for quickly breeding, the tissue that relates in particular to a kind of Agrimony is cultivated and method for quickly breeding.
Background technology
Agrimony (
Agrimonia pilosa) belong to herbaceos perennial for the rose family (Rosaceae) Agrimony, be a kind of important medicinal plant, its acrial part, all can be used as medicine with hibernaculum and the root of short and small rhizome.Wherein, acrial part is used as medicine and claims hairyvein agrimony, different name fortune silene herb with root, wolf's fang grass, goldentop fossilia dentis mastodi, hairyvein agrimony, Agrimony, Huanglong's tail, triflorous ainsliaea leaf, dry foot mattress etc., property is put down, bitter, puckery, return lung, liver, the spleen channel, Bearberry Extract hemostasis, anti-inflammtory anti-dysentery, desinsection and antitumor effect, the clinical infectious food poisoning of Halophiles, trichomonas vaginitis and the meniere's syndrome of being used for the treatment of very imitated; Be used as medicine with the hibernaculum of short and small rhizome and claim gemma agrimoninae, different name wolf's fang, wolf tooth, wolf's fang, wolf's fang grass roots bud, hairyvein agrinonia rhizome etc., it is cool in nature, bitter, puckery, the effect of tool expelling parasite and removing toxicity for detumescence cures mainly tapeworm disease, trichomoniasis, sore mange, furuncle and dysentery.Clinical treatment tapeworm disease has good effect; A root hairyvein agrimonia root that is used as medicine, different name frozen ground wind, it is warm in nature, and flavor is hot, puckery, and the effect of tool detoxifcation and expelling parasite cures mainly the diseases such as dysentery, sore, pyogenic infections, malaria, tapeworm disease and amenorrhoea.In the last few years, chemical composition analysis, extraction separation, assay method and the effect of Agrimony there had been more deep research.The breeding of present imperial bud adopts seed and root division mode to carry out, and these two kinds of methods exist floods the shortcoming that reproduction coefficient is low, the breeding cycle is long, therefore sets up the tissue-culturing rapid propagation system of Agrimony, is the another effective way that realizes that its industrialization is produced.
Report (" Plant Physiology Communications " 04 phase in 1986 that existing Agrimony tissue is cultivated, the tissue of Agrimony is cultivated, Wu Meifen, Chen Weidong): the adventitious organogenesis take hypocotyl, cotyledon and the blade of Agrimony seedling as explant, blade and hypocotyl need through 2, in the 4-D evoked callus stage, cotyledon can differentiate indefinite bud behind MS+6-BA 2.0 mg/L NAA 0.2 mg/L evoked callus.
Summary of the invention
In order to solve above-mentioned technical problem, the tissue that the purpose of this invention is to provide a kind of Agrimony is cultivated and method for quickly breeding, the tissue that adopts the method to carry out Agrimony is cultivated and Fast-propagation, improved rooting rate and the number of taking root, and length and the rugosity of root are moderate, have greatly improved the survival rate of transplanting.
In order to realize above-mentioned purpose, the present invention has adopted following technical scheme:
The tissue of Agrimony is cultivated and method for quickly breeding, and the method comprises the steps:
1) sterilization of explant:
Get the larger leaflet in the firm fully-developed Agrimony winglike compound leaf, add washing agent and carefully scrub, wash with flowing water again, in super-clean bench, blade changed in the alcoholic solution and soak, after change soaking disinfection in the thimerosal over to, use aseptic water washing, then blade is cut into 1 ~ 3 cm
2Fritter, be inoculated on the MS medium that adds different plant growth regulating substances with heads horizontal disposing way;
2) adventitious bud inducing
The composition of the plant growth regulating substance that adds in the MS medium of explant access comprises:
6-BA 0.5~5.0 mg/L
TDZ 0.1~1.0 mg/L；
Secretly cultivate first, temperature is 25 ± 1 ℃, and condition of culture is light/dark hour than being 12 ~ 16:8 ~ 12, intensity of illumination 30 ~ 40 μ mol/ms;
3) cut the indefinite bud that blade is light green, growth is vigorous and 2 ~ 3 cm are high, be inserted in the root media, comprise in the described root media:
Minimal medium (1/4 ~ 1) MS
NAA 0.05~0.5 mg/L；
Crack cultivation bottle cap carries out hardening during hardening, and the medium that taking-up and clean root adhere to behind 20 ~ 30h moves in the soil, and the initial stage will be carried out moist keeping measures, and In Shade, treats that young leaves grows, and can remove beaker.
As preferably, above-mentioned step 1) thimerosal is for dripping the 0.2%HgCl of 1 a mol/L NaOH
2Solution.Be 10 ~ 11 min as preferred again, above-mentioned disinfecting time.
The composition of the plant growth regulating substance that adds as preferably, above-mentioned step 2) comprises:
6-BA 1.0~4.0 mg/L
TDZ 0.1~0.8mg/L。
As most preferably, above-mentioned step 2) plant growth regulating substance that adds in is selected TDZ 0.3 mg/L+6-BA 2.0 mg/L.
As preferably, above-mentioned step 2) composition of plant growth regulating substance also comprises IAA, NAA or both combinations, and the concentration of IAA is 0 ~ 2.0 mg/L, and the concentration of NAA is 0 ~ 0.5 mg/L.
As preferred, above-mentioned again step 2) composition of plant growth regulating substance also comprises IAA, the concentration of IAA is 0.5 ~ 1.5 mg/L.
As preferably, NAA is 0.08 ~ 0.3mg/L in the above-mentioned step 3) root media.
As most preferably, above-mentioned step 3) root media is 1/4MS+NAA 0.1 mg/L.
The present invention has been owing to having adopted above-mentioned technical scheme, and that adopts that the method carries out Agrimony has improved rooting rate and the number of taking root with Fast-propagation, and the length of root and rugosity are moderate, have greatly improved the survival rate of transplanting.The method has been set up the tissue-culturing rapid propagation system of Agrimony, is the effective way that realizes that its industrialization is produced.
Embodiment
Material source: Agrimony is picked up from Lishui White Cloud Mountain, transplants and plants to China Measures Institute solarium.
Explant material: the blade of Agrimony.
1. the sterilization of explant: get the larger leaflet in the firm fully-developed Agrimony winglike compound leaf, adding an amount of washing agent carefully scrubs, again with behind flowing water flushing 0.5 ~ 1.0 h, in super-clean bench, blade changed in 70% the alcoholic solution and soaks 30 s, after change the 0.2%HgCl that has dripped 11 mol/L NaOH over to
2The soaking disinfection some time in the solution, use aseptic water washing 4 ~ 5 times, then blade is cut into 1 ~ 3 cm
2Fritter, be inoculated on the MS medium that adds different plant growth regulating substances with heads horizontal disposing way.
1.1 result: sterilization 5 ~ 6 min reach optimum efficiency, and the pollution rate of explant and lethality all are lower than 10%.
2. adventitious bud inducing preliminary experiment
The composition of the plant growth regulating substance that adds in the MS medium of explant access is respectively:
1) 2,4-D 2.0 mg/L +6-BA 0.5 mg/L；
2) 6-BA 2.0 mg/L +NAA 0.1 mg/L；
3) TDZ 0.3 mg/L
-1+NAA 0.1 mg/L；
4) TDZ 0.05 mg/L +2,4-D 2.0 mg/L；
5) TDZ 0.3 mg/L +6-BA 2.0 mg/L。
Repeat 3 times.Sucrose concentration is 3% in all medium, and agar concentration is 0.7%, pH 5.8-6.0, and is lower same.Secretly cultivate first, temperature is (25 ± 1) ℃, and condition of culture is that 14h light/10h is dark, intensity of illumination 30 ~ 40 μ mol/ms, lower with.Observe at ordinary times explant growth situation and record, extract 10 bottles out and add up at random when 30 d, and filter out better plant growth regulating substance and make up to be optimized.
2.1 result: add 2, do not induce indefinite bud in the medium of 4-D (1 and No. 4 medium), and all induced indefinite bud in all the other 3 kinds of medium, but the time that indefinite bud occurs differs, to add TDZ 0.3 mg/L+6-BA 2.0 mg/L the earliest (No. 5 medium), the earliest can 12 d, inductivity is the highest (65.6%) also, add 6-BA 2.0 mg/L+NAA 0.1 mg/L the latest (No. 2 medium), earliest time is 20 d, inductivity is minimum (12.2%) also, therefore select TDZ and 6-BA as the candidate plant growth regulating substance in the screening of best adventitious bud inducing.Simultaneously, in incubation, observe, only observe less more tissue in the medium of interpolation 2,4-D, and in the medium that produces indefinite bud, its part leaf dish base portion expands after finding certain hour, the edge perk, and the paddle cutout place has the green matter of similar callus to form, and base portion is more than the end, but carefully be distinguished as the budlet point, and more obvious along with the increase of incubation time, the maximum part of indefinite bud occurring and be leave handle place.
3. the optimization of the plant growth regulating substance of adventitious bud inducing combination
TDZ (0.1,0.3 and 1.0 mg/L) and 6-BA (0.5, the 2.0 and 5.0 mg/L) combination of 3 gradients respectively are set, and preparation MS cultivates, and with the explant inoculation thereon, 30 d extract 10 bottles out at random and add up.
The rising of 3.1 result: TDZ and 6-BA concentration, adventitious bud induction frequency shows zooming trend with inducing to count, but descends to some extent when high concentration, and each inductivity of processing is between 44.1 ~ 8.9%, and the indefinite bud quantity that induces is between 3.3 ~ 7.7.The synergistic effect of two plant growth regulating substances is also very obvious, especially during low concentration, the raising of a plant growth regulating substance concentration can improve fast inductivity and induce number, adventitious bud induction frequency to reach as high as about 90%, and the indefinite bud that induces also has about 8.But the TDZ of high concentration and 6-BA have brought vitrified problem, and both also have synergistic effect, the TDZ of maximum concentration and 6-BA combination, and its vitrifying is the most serious.The vitrifying degree of moderate TDZ and 6-BA is lower, and adventitious bud induction frequency and induce the number higher, especially good with being combined as of TDZ 0.3 mg/L+6-BA 2.0 mg/L, inductivity can arrive 85.6%, indefinite bud quantity can reach 7.4, and well-grown.
4. different growth regulatory substance combinations are on the impact of proliferate
Though MS+TDZ 0.3 mg/L+6-BA 2.0 mg/L can induce indefinite bud, though find that in subculture indefinite bud has propagation, poor growth, seldom be higher than 2 cm during 30 d, the experiment that is difficult to take root is therefore in proliferate is cultivated, additionally add growth hormone IAA and NAA again and screen.The indefinite bud clump that will in MS+TDZ 0.3 mg/L+6-BA 2.0 mg/L medium, induce, be cut into the fritter of 2 ~ 3 strains, variable concentrations IAA (0,0.2,0.5,1.0 and 2.0 mg/L) or NAA (0,0.05,0.1,0.2 and 0.5 mgL are additionally added in inoculation again
-1) Optimal Medium in.30 d extract 10 bottles out at random and add up.
4.1 result: along with the rising of IAA concentration, the rate of increase of indefinite bud improves, and growth is also accelerated simultaneously, and reach stable at 1 mg/L, the rate of increase can be elevated to 4.39 from 4.03, but NAA is all little to the rate of increase and indefinite bud affects on the growth, between 4.09 ~ 4.14.
5. the culture of rootage of regeneration plant and transplanting
Cut the indefinite bud that blade is light green, growth is vigorous and 2 ~ 3 cm are high, be inserted in the root media, minimal medium (MS, 1/2MS and 1/4MS), NAA (0,0.1 and 0.5 mg/L) and the inositol (0%, 50%, 100%) of 3 gradients respectively are set, extract 10 bottles out at random during 30 d and add up rooting rate.Can carry out hardening by crack cultivation bottle cap during hardening, the medium that taking-up and clean root adhere to behind 1 d moves in the soil, and soil is little silt, and the initial stage will be carried out moist keeping measures, and In Shade, treats that young leaves grows, and can remove beaker, adds up simultaneously transplanting survival rate.
5.1 result: Agrimony takes root easily, and each rooting rate of processing is between 88 ~ 98%.Along with MS concentration reduces, rooting rate improves, and the radical amount increases, and is elongated, attenuates; And along with the increase of NAA concentration, taking root takes the lead in raising descends again, and the quantity of root increases first and reduces, and the length of root also shows and increases first the trend that reduces afterwards, but the rugosity of root shows the trend of increase; But the concentration of inositol is little for the culture of rootage impact.In addition, observe to find, each rootage duration of processing is close, with 1/4MS+NAA 0.1 mg/L the earliest (6 d), with MS the latest (8 d).But have any different on the transplanting survival rate, when high concentration (0.5 mg/L) NAA cultivates, although the quantity of root increases, but elongated, shorten, the lignification of these roots is obvious, root becomes thick and hard, transplanting survival rate (81%) reduces on the contrary, and significantly is lower than other processing; No significant difference between the transplanting survival rate of the different regrowths that obtain of MS concentration and inositol concentration all reaches about 95%.Therefore comprehensive rooting rate, radical, root are grown, root is thick and transplanting survival rate, suitable root media is 1/4MS+NAA 0.1 mg/L, if consideration cost factor, can not add inositol, 1/4MS+NAA 0.1 mg/L that does not namely add inositol is best root media, and taking root to reach 98%, and the number of taking root is 6.1, length and the rugosity of root are moderate, and transplanting survival rate can reach 96%.
Claims (9)
1. the tissue of Agrimony is cultivated and method for quickly breeding, it is characterized in that the method comprises the steps:
1) sterilization of explant:
Get the larger leaflet in the firm fully-developed Agrimony winglike compound leaf, add washing agent and carefully scrub, wash with flowing water again, in super-clean bench, blade changed in the alcoholic solution and soak, after change soaking disinfection in the thimerosal over to, use aseptic water washing, then blade is cut into 1 ~ 3 cm
2Fritter, be inoculated on the MS medium that adds plant growth regulating substance with heads horizontal disposing way;
2) adventitious bud inducing
The composition of the plant growth regulating substance that adds in the MS medium of explant access comprises:
6-BA 0.5~5.0 mg/L
TDZ 0.1~1.0 mg/L；
Secretly cultivate first, temperature is 25 ± 1 ℃, and condition of culture is light/dark hour than being 12 ~ 16:8 ~ 12, intensity of illumination 30 ~ 40 μ mol/ms;
3) cut the indefinite bud that blade is light green, growth is vigorous and 2 ~ 3 cm are high, be inserted in the root media, comprise in the described root media:
Minimal medium (1/4 ~ 1) MS
NAA 0.05~0.5 mg/L；
Crack cultivation bottle cap carries out hardening during hardening, and the medium that taking-up and clean root adhere to behind 20 ~ 30h moves in the soil, and the initial stage will be carried out moist keeping measures, and In Shade, treats that young leaves grows, and can remove beaker.
2. the tissue of Agrimony according to claim 1 is cultivated and method for quickly breeding, it is characterized in that the step 1) thimerosal is for dripping the 0.2%HgCl of 1 a mol/L NaOH
2Solution.
3. the tissue of Agrimony according to claim 2 is cultivated and method for quickly breeding, it is characterized in that the step 1) disinfecting time is 5 ~ 6 min.
4. the tissue of Agrimony according to claim 1 is cultivated and method for quickly breeding, it is characterized in that step 2) in the composition of the plant growth regulating substance that adds comprise:
6-BA 1.0~4.0 mg/L
TDZ 0.1~0.8mg/L。
5. the tissue of Agrimony according to claim 1 is cultivated and method for quickly breeding, it is characterized in that step 2) in the plant growth regulating substance that adds select TDZ 0.3 mg/L+6-BA 2.0 mg/L.
6. according to claim 1 or the tissue of 3 or 4 described Agrimonies is cultivated and method for quickly breeding, it is characterized in that step 2) composition of plant growth regulating substance also comprises IAA, NAA or both combinations, the concentration of IAA is 0 ~ 2.0 mg/L, and the concentration of NAA is 0 ~ 0.5 mg/L.
7. the tissue of Agrimony according to claim 5 is cultivated and method for quickly breeding, it is characterized in that step 2) composition of plant growth regulating substance also comprises IAA, the concentration of IAA is 0.5 ~ 1.5 mg/L.
8. the tissue of Agrimony according to claim 1 is cultivated and method for quickly breeding, it is characterized in that NAA is 0.08 ~ 0.3mg/L in the step 3) root media.
9. the tissue of Agrimony according to claim 7 is cultivated and method for quickly breeding, it is characterized in that the step 3) root media is 1/4MS+NAA 0.1 mg/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210505869.2A CN103004589B (en) | 2012-12-03 | 2012-12-03 | Tissue culture and rapid propagation method of agrimony |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210505869.2A CN103004589B (en) | 2012-12-03 | 2012-12-03 | Tissue culture and rapid propagation method of agrimony |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103004589A true CN103004589A (en) | 2013-04-03 |
| CN103004589B CN103004589B (en) | 2014-08-06 |
Family
ID=47954120
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210505869.2A Expired - Fee Related CN103004589B (en) | 2012-12-03 | 2012-12-03 | Tissue culture and rapid propagation method of agrimony |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103004589B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104813940A (en) * | 2015-05-18 | 2015-08-05 | 中国计量学院 | Rapid asystasiella neesiana tissue culture propagation method |
| CN105961202A (en) * | 2016-05-26 | 2016-09-28 | 上海市园林科学规划研究院 | Rapid propagation method of Agrimonia pilosa |
| CN114766319A (en) * | 2017-11-24 | 2022-07-22 | 浙江省农业科学院 | Strawberry cultivation medium and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000057690A2 (en) * | 1999-03-25 | 2000-10-05 | University Of Guelph | Micropropagation and production of phytopharmaceutical plants |
| CN101268758A (en) * | 2008-05-09 | 2008-09-24 | 南京林业大学 | Quick replication method for Xuan pawpaw tissue cultivation |
-
2012
- 2012-12-03 CN CN201210505869.2A patent/CN103004589B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000057690A2 (en) * | 1999-03-25 | 2000-10-05 | University Of Guelph | Micropropagation and production of phytopharmaceutical plants |
| CN101268758A (en) * | 2008-05-09 | 2008-09-24 | 南京林业大学 | Quick replication method for Xuan pawpaw tissue cultivation |
Non-Patent Citations (1)
| Title |
|---|
| 吴美芬: "龙芽草的组织培养", 《植物生理学通讯》, no. 4, 15 April 1986 (1986-04-15), pages 56 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104813940A (en) * | 2015-05-18 | 2015-08-05 | 中国计量学院 | Rapid asystasiella neesiana tissue culture propagation method |
| CN105961202A (en) * | 2016-05-26 | 2016-09-28 | 上海市园林科学规划研究院 | Rapid propagation method of Agrimonia pilosa |
| CN114766319A (en) * | 2017-11-24 | 2022-07-22 | 浙江省农业科学院 | Strawberry cultivation medium and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103004589B (en) | 2014-08-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107047320B (en) | A kind of bigflower centranthera root method for tissue culture | |
| CN102204512B (en) | Tissue culture method for lilium tenuifolium | |
| CN104663450A (en) | Tissue culture and rapid propagation method for Acer rubrum 'Brandywine' | |
| CN1748478A (en) | Method for cultivating and producing detoxic seedling on strawberry stemp apex | |
| Martin et al. | An efficient micropropagation system for Celastrus paniculatus Willd.: a vulnerable medicinal plant | |
| CN106665357A (en) | Method for establishing lycoris regeneration system | |
| US11547071B2 (en) | Methods for disinfecting and inducing direct rapid proliferation of explants of Kadsura coccinea stems with buds | |
| CN103314863B (en) | A kind of method of efficient acquisition tulip callus | |
| CN100512631C (en) | Method and technique of the separated tissue culture of Kava | |
| CN103004589B (en) | Tissue culture and rapid propagation method of agrimony | |
| CN103004588B (en) | Tissue culture and rapid propagation method of tulipa edulis | |
| CN104094845B (en) | A kind of in-vitro culture method of Dendranthema indicum | |
| Abhyankar et al. | Rapid micropropagation via axillary bud proliferation of Adhatoda vasica Nees from nodal segments | |
| CN112470929B (en) | Method for obtaining regeneration plant from root-neck apical tissue of rhodiola crenulata | |
| CN109220804A (en) | A kind of high efficiency quick breeding method of the thick rib grass of coloured silk leaf | |
| CN101015280B (en) | Tissue culture method for fast propagation of primula denticulata ssp.sino-denticulata | |
| CN102396418A (en) | Iris germanica Jinwawa tissue culture method | |
| Sharma et al. | In vitro propagation of Citrus rootstocks | |
| CN104429940B (en) | A kind of method obtaining Fructus Fragariae Ananssae detoxic seedling | |
| Mishra et al. | Micropropagation of guava (Psidium guajava L.) | |
| CN105532452A (en) | Method for inducing and quickly breeding daphnemezereum seeds | |
| CN106613933A (en) | Method for inducing rooting of apple dwarf rootstock tissue culture seedling | |
| CN105519435B (en) | A kind of tissue culture method of induction fritillaria thunbergii clove | |
| CN105766643A (en) | Method for obtaining explant based on heading-back twig in August to September to increase survival rate of Dangshan crisp pear tissue culture seedlings | |
| CN106386492B (en) | Induction method of loose embryonic callus of potato |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140806 Termination date: 20171203 |