CN103444530B - Technology for field return-to-nature ex-situ plantation of wild anoectochilus roxburghii (Wall.) Lindl in Fujian - Google Patents
Technology for field return-to-nature ex-situ plantation of wild anoectochilus roxburghii (Wall.) Lindl in Fujian Download PDFInfo
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Abstract
The invention discloses a technology for field return-to-nature ex-situ plantation of wild anoectochilus roxburghii (Wall.)Lindl in Fujian. The technology comprises the steps of obtaining a material in the field, disinfecting the material, performing induction culture on adventitious buds, performing one-time grown-up seedling culture, transplanting bottle seedlings, performing underwood planting, and preparing a finished product. The technology solves the problems that the seeds of anoectochilus roxburghii (Wall.) Lindl are tiny, embryos are not completely developed, and the seeds are difficult to germinate under a natural condition; the method also solves the problems that in the prior art, a medicinal herb resource of the anoectochilus roxburghii (Wall.) Lindl is rare, and especially, the components for strongly easing pain, diminishing inflammation and resisting cancer contained in plants can not meet clinical requirements and daily requirements for enhancing human body immunity and the like. According to the technology, the wild anoectochilus roxburghii (Wall.) Lindl is cultured by a one-time grown-up seedling method and is subjected to field return-to-nature ex-situ plantation, so that the survival rate of the anoectochilus roxburghii (Wall.) Lindl is high, the yield is improved by 1-2 times, and the plants contain sufficient substances and are good in condition and quality.
Description
Technical field
The invention belongs to field of plant tissue culture technique, be specifically related to a kind of Wild in Fujian Province roxburgh anoectochilus terminal bud field naturally return move ground planting technology.
Background technology
Roxburgh anoectochilus terminal bud (
anoectochilus roxburghii(Wall.) Lindl) be one of precious herbal medicine, belong to the orchid family (
orchidaceae) Anoectochilus Blume (
anoectochilus) plant, also known as gold thread city, bird ginseng, enjoy the laudatory title such as " king of medicine ", " refreshing medicine ".Roxburgh anoectochilus terminal bud is Lu Shenglan, perennial herb, high 4 ~ 18cm; Root-like stock crawls to extend, reddish tan, slightly meat, long 4 ~ 9cm, and tool obviously saves, and long on joint have adventive root; Leaf alternate, width egg shape is oval to ovum shape, long 1.5 ~ 3.5cm; wide 1 ~ 3cm; vein is netted, and golden yellow, leaf lower surface is lilac red; primary growth is under height above sea level 300 ~ 600 meters of remote, thickly forested mountains or in the other moist thick grass in mountain stream or under bamboo grove; shady wetland band particularly under broad leaved forest, natural environment growth roxburgh anoectochilus terminal bud generally in sparse, sporadicly distribute, little close life in blocks; germ plasm resource is rare, is imminent extinction protected plants.Roxburgh anoectochilus terminal bud seed is small, and embryonic development is incomplete, difficulty of germinateing under field conditions (factors).Mainly be distributed in the states such as the Japan in Asia, China, Sri Lanka, India and Nepal, Taiwan and Fujian are one of main places of production of China.
Roxburgh anoectochilus terminal bud is containing carbohydrate, taurine, Cardiac glycosides, ester class, alkaloid, steroidal, several amino acids, trace element and inorganic elements etc.There are some researches show, roxburgh anoectochilus terminal bud has and improves immunity of organisms, the effect such as anti-ageing, hypoglycemic.Wherein, anoectochilus roxburghii polyose is very promising antineoplastic or assistant anti-tumor drug.Extracting substance in the plant of roxburgh anoectochilus terminal bud, can successfully suppress the most ticklish breast cancer (MCF-7), accelerates and induced breast cancer cell death, and block cancer cell structural generation, this extract also has fungistatic effect, can kill common bacterial infection.Various places, the Fujian people, roxburgh anoectochilus terminal bud asks that medicining condition is slightly different: Minxi region children's's high fever, the infantile convulsion etc. of being used among the people such as (peaceful north, clear stream, Taining county); South Fujian Province is used for treatment ephritis, cystitis, diabetes etc.; Area, the middle part of Fujian Province is used for anti-inflammatory analgesic, treatment gall stone, cholangitis etc.; Area, east, Fujian is used for wines used as antirheumatic.Stimulate the menstrual flow network, nourish blood, logical numbness, treatment rheumatism joint is sick, waist knee numbness pain etc.
At present, roxburgh anoectochilus terminal bud industrialization begins to take shape.Mainly be distributed in Fujian, the environment such as Guangdong are comparatively applicable to the area of its growth, and annual value of production is more than 1,000,000,000 yuan of high pointes.But in roxburgh anoectochilus terminal bud industrialization production process, be subject to the restriction of nutriment in bottle, somatotropin content, growing space, cause the content of some nutriments in its plant corpus lower than the roxburgh anoectochilus terminal bud under wild environment.Most enterprise is when setting up roxburgh anoectochilus terminal bud production line, and plantation is main does not carry out rational underwood planting based on greenhouse gardening, and without ripe complete large-scale production system, the quality of producing roxburgh anoectochilus terminal bud is uneven.Therefore a roxburgh anoectochilus terminal bud industrialization standard is built, set up a complete roxburgh anoectochilus terminal bud production system pattern, " roxburgh anoectochilus terminal bud underwood planting " technology of exploitation, and accelerate its industrialization to the raising people's livelihood general level of the health, promote that the health preserving health-care food of distinct Chinese characteristics has important practical significance.
Summary of the invention
The object of the invention one is to provide a kind of application one-step-seedling formation culture technique, moves ground growing and cultivation roxburgh anoectochilus terminal bud promote that industry development provides key technology for sylvan life.Two is that solution roxburgh anoectochilus terminal bud seed is small, and embryonic development is incomplete, difficult problem of germinateing under field conditions (factors); The roxburgh anoectochilus terminal bud medicine source in prior art that solves is rare, and particularly contained in plant strong analgesia, anti-inflammatory, anticancer component can not meet the problems such as clinical and daily enhancing body immunity needs.
Method of the present invention realizes as follows:
A kind of Wild in Fujian Province roxburgh anoectochilus terminal bud field naturally returns moves planting technology, it is characterized in that comprising draw materials, prepared by materials disinfection process, adventitious bud induction culture, one-step-seedling formation cultivation, bottle transplantation of seedlings, underwood planting, finished product.Concrete steps are as follows:
1) draw materials: wild roxburgh anoectochilus terminal bud plants stems section, pick up from Yongtai, Fujian height above sea level 300 ~ 600m mountain stream, wrap with wet cloth after plucking and take back;
2) materials disinfection process: by clean for the wild roxburgh anoectochilus terminal bud tap water gathered, in distance bud point 1 ~ 2cm place clip stem section, remove unnecessary blade, put rinsing 15min in saturated bleaching powder supernatant and scrub gently with banister brush, after rinsing well, being placed in flowing water punching and dripping 0.5 ~ 1 hour, rinse 2 ~ 3 times with distilled water again, with the alcohol disinfecting 30s of 75% in superclean bench, add 0.1% mercuric chloride process 5 ~ 10min, more for subsequent use with sterile water punching 3 ~ 4 times rearmounted workbench;
3) adventitious bud induction culture: the material after sterilization is placed on sterilized culture dish, cut out portion 0.2 ~ the 0.4cm contacted with mercuric chloride is cut away with scalpel, then stem section material being cut into 1 ~ 2cm long is inoculated in respectively in high temperature, autoclaved adventitious bud induction culture base, cultivation temperature is 23 ± 2 DEG C, light application time 10h/d, cultivates and carries out black dim cultivation in first 10 days, and after 11 days, intensity of illumination is adjusted to 500-1000lx, bud inducement is cultivated 35 ~ 45 days, obtains Multiple Buds;
4) one-step-seedling formation is cultivated: will obtain Multiple Buds through Fiber differentiation, high 3 ~ 4cm, tool 3 ~ 4 internode, the unrooted seedling of 1 ~ 2 leaf, the stem section being cut into 1 ~ 2cm long is inoculated in high temperature, autoclaved one-step-seedling formation medium, and stem section must tile storing, cultivation temperature is 23 ± 2 DEG C, light application time 12h/d, intensity of illumination is 1500 ~ 2500lx, cultivate within 4 ~ 5 months, obtain the complete and rooting rate of growth reach 99% can directly transplanting bottle seedling plant;
5) bottle transplantation of seedlings: synchronously will to take root and plant height is the roxburgh anoectochilus terminal bud bottle seedling of 6 ~ 10cm through one-step-seedling formation Fiber differentiation 4 ~ 5 months, be placed on natural daylight lower refining seedling 3 ~ 5 days, open bottle cap hardening again 2 ~ 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: select that height above sea level 300 ~ 800 meters of soil properties are loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, transplant front matrix 45% carbendazol wettable powder, 900 times of solution and irrigate sterilization; During plantation, every seeding row spacing 5*5cm, the cultivation height of bed 20 ~ 30cm, the wide 1 ~ 1.2m of bed surface, density 500 ~ 1000 strains/m
2; During cultivation, keep ground moistening and temperature 12 ~ 15 DEG C, apply fertilizer the roxburgh anoectochilus terminal bud season of growth every two weeks use farmers''s liquid fertilizer or gas-liquid, be diluted to 1000 times of uses; Sylvan life nature cultivation 6 ~ 8 months, plant height 10 ~ 15cm, can gather when growing 5 ~ 8 leaves, is 99% by the survival rate of underwood planting roxburgh anoectochilus terminal bud;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can make fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dry or dry product three kinds.
Wherein, the formula of described step 3) adventitious bud induction culture base is MS+BA 2mgL
-1+ NAA 1mgL
-1+ KT 0.5mgL
-1+ N6-isopentene base adenine 1mgL
-1+ sucrose 30gL
-1+ active carbon 1gL
-1+ agar 6.5gL
-1, medium thickness is 1.4 ~ 1.6cm.
The formula of described step 4) one-step-seedling formation medium is 1/3MS+N6-isopentene base adenine 0.5 ~ 1.5mgL
-1+ NAA 1 ~ 2mgL
-1+ HyponexI 2 ~ 3gL
-1+ Hyponex2 2 ~ 3gL
-1+ Peptone 1 ~ 3gL
-1+ potato 100 ~ 150gL
-1+ sucrose 35gL
-1+ active carbon 1 ~ 2gL
-1+ agar 6.5gL
-1, medium thickness is 1.4 ~ 1.6cm.
The present invention has following remarkable advantage:
1, apply one-step-seedling formation technology groups training roxburgh anoectochilus terminal bud, improve roxburgh anoectochilus terminal bud transplanted seedling survival rate and rooting rate.Naturally returned by sylvan life and plant with moving, the content of some hormones of release incubation, improve the growing ability of roxburgh anoectochilus terminal bud and the accumulation of benefit materials further, after plantation, roxburgh anoectochilus terminal bud increases by 1 ~ 2 times than bottle transplantation of seedlings phase weight.
2, the present invention sets up one-step-seedling formation culture technique, transplanting and simulation sylvan life growth; naturally a complete set of production technology system of planting with moving is returned; for the formation of the industrialization of whole roxburgh anoectochilus terminal bud industry, scale, standardization cultivation system provides technical support; simultaneously guaranteeing the quality, on the basis of guarantor's amount; accelerate capital fund collection process; meet the market demand, investment reduction enterprise plenty of time and fund.
3, economize the land resource; protection peasant benefit; in agricultural sector structure adjustment; make full use of forest land resource; development is applicable to the Wild in Fujian Province roxburgh anoectochilus terminal bud of sylvan life growth, and such peasant's vested interest is without prejudice, and does not affect not only agricultural production simultaneously but also create the agricultural products of high added value; add farmers' income, truly accomplish and reach the aim of new and high technology serving agriculture, Serve the farmers.
4, utilize forest land resource, increase economic efficiency.Give full play to the resourceful advantage of forest land of mountain area, improve forest land utilization rate, greatly improve the economic benefit of forest, for the reasonable arrangement of mountain area labour provides condition, the labour simultaneously for recurrence and abundant current mountain area is best appeal.
5, forest ecology protection is conducive to, in conjunction with fostering, playing branch, topdress, promote tree growth.
6, sylvan life is carried out artificial cultivation and can be met growing environment needed for roxburgh anoectochilus terminal bud wild state, saves the input of sunshade net simultaneously.Therefore, these species of protection roxburgh anoectochilus terminal bud have important scientific research, medical science, ecological significance and value, will play an important role for mankind's treating disease and health.Carrying out calculating with 6 months according to current plantation situation is a production cycle, develop at a progressive speed, one mu of mountain region is planted every year can twice harvest, by the fresh goods 300 yuan/kg of (current market price) underwood planting, every mu of results fresh goods 500kg*2, every mu of annual value of production 600000 RMB, so Anoectochilus roxburghii underwood planting has a extensive future.
The present invention bases oneself upon Fujian mountain area forest land resource advantage, " carry out forestry specialty, ecological cultivation " principle, adopt sylvan life to return to roxburgh anoectochilus terminal bud to cultivate with moving, build the optimum growh environment being applicable to roxburgh anoectochilus terminal bud bottle seedling, improve transplanting success, form the scale of roxburgh anoectochilus terminal bud plantlet in vitro underwood planting, set up cultivation system under the forest plantation of Anoectochilus roxburghii, improve roxburgh anoectochilus terminal bud Industry Promotion system so neither to take peasant and can plough soil, can preserve the ecological environment again, through for many years, multiple spot experiment in cultivation, development Anoectochilus roxburghii industry, sylvan life exploitation must be carried out, implement ecological cultivation, reach the effect of economic benefit and ecological benefits doulbe-sides' victory.
Accompanying drawing explanation
Fig. 1 one-step-seedling formation cultivates the growing state that tiling is placed
The transplant of Fig. 2 batch cultur
Fig. 3 prepares the large measuring bottle seedling transplanted
Fig. 4 is bottle seedling growing state after transplanting
Fig. 5 underwood planting habitat
Fig. 6 underwood planting seedling
Fig. 7 underwood planting 6 months growing states
The roxburgh anoectochilus terminal bud that Fig. 8 has been dried
Fig. 9 finished product roxburgh anoectochilus terminal bud
Embodiment
In order to fully disclose Wild in Fujian Province roxburgh anoectochilus terminal bud field of the present invention naturally return move ground planting technology, the present invention is described further below, but the present invention is not limited thereto.
embodiment 1
1) draw materials: wild roxburgh anoectochilus terminal bud plants stems section, pick up from Yongtai, Fujian height above sea level 500m mountain stream, wrap with wet cloth after plucking and take back.
2) materials disinfection process: by clean for the wild roxburgh anoectochilus terminal bud tap water gathered, in distance bud point 2cm place clip stem section, remove unnecessary blade, put rinsing 15min in saturated bleaching powder supernatant and scrub gently with banister brush, after rinsing well, being placed in flowing water punching and dripping 1 hour, 3 times are rinsed again with distilled water, with the alcohol disinfecting 30s of 75% in superclean bench, add 0.1% mercuric chloride process 8min, then it is for subsequent use to rush 4 times rearmounted workbench with sterile water.
3) adventitious bud induction culture: the material after sterile-processed is placed on sterilized culture dish, cut away the cut out portion 0.2cm contacted with mercuric chloride with scalpel, the stem section then material being cut into 2cm long is inoculated in respectively in high temperature, autoclaved adventitious bud induction culture base.The formula of medium is MS+BA 2mgL
-1+ NAA 1mgL
-1+ KT 0.5mgL
-1+ N6-isopentene base adenine 1mgL
-1+ sucrose 30gL
-1+ active carbon 1gL
-1+ agar 6.5gL
-1, medium thickness is 1.5cm.
Cultivation temperature is 23 DEG C, light application time 10h/d, cultivates and carries out black dim cultivation in first 10 days, and after 11 days, intensity of illumination is adjusted to 800lx, and bud inducement cultivates 40 days, obtains Multiple Buds.
4) one-step-seedling formation is cultivated: will cultivate through bud inducement and obtain Multiple Buds, high 3cm, tool 3 internode, the unrooted seedling of 2 leaves, and be cut into the stem section that 2cm grows and be inoculated in high temperature, autoclaved one-step-seedling formation medium, stem section must tile storing, sees Fig. 1.The formula of medium is 1/3MS+N6-isopentene base adenine 1.0mgL
-1+ NAA 1.5mgL
-1+ HyponexI 2.5gL
-1+ Hyponex2 2.5gL
-1+ Peptone 2gL
-1+ potato 120gL
-1+ sucrose 35gL
-1+ active carbon 1.5gL
-1+ agar 6.5gL
-1, medium thickness is 1.5cm.
Cultivation temperature is 23 DEG C, light application time 12h/d, and intensity of illumination is 2000lx, cultivates within 4.5 months, to obtain that growth is complete and rooting rate reaches 99% directly can transplant a bottle seedling plant, sees Fig. 2, Fig. 3.
5) bottle transplantation of seedlings: synchronously will to take root and plant height is the roxburgh anoectochilus terminal bud bottle seedling of 8cm through one-step-seedling formation Fiber differentiation 4.5 months, be placed on natural daylight lower refining seedling 5 days, then open bottle cap hardening 2 days, see Fig. 4.Then from blake bottle, take out plant, wash away the medium be attached on root system.
6) underwood planting: comprise select with planting, clear up with planting, put up shade, matrix is disinfected, underplanting, manage after transplanting, sylvan life gathers.
A) select with planting
Select that height above sea level 500 meters of soil properties are loose, ventilative, draining, mountain region that water retention property is good, humidity should remain between 70%-85%, avoids direct sunlight, light transmittance 50%-60%, and suitable to give birth to temperature be 15-28 DEG C, and roxburgh anoectochilus terminal bud should grow under Acid soil condition.
B) clear up with planting
Land readjustment, weed with a hoe, the shrub of surrounding and some tree branches are carried out between dredge, surgery, make flat simultaneously, the wide 30-60 centimetre in the ridge, high 20 centimetres, also can plant position according to mountain region actual and to arrange and tree line-spacing is grasped flexibly.Under field conditions (factors), upper layer trees crown density 65%-75%, low cover cover degree 35-45%.
C) shade is put up
Put up shade, regulate light transmittance, printing opacity is applicable with three sun seven the moon, and 4 ~ May, temperature raised, and printing opacity can be reduced to less than 30%, must adopt fences basketry, draws lid sunshade net to prevent the invasion and attack of the wildlife harm such as mouse, snake, bird, worm around planting site.
Fig. 5 is seen in underwood planting habitat.
D) matrix is disinfected
Before transplanting, matrix 45% carbendazol wettable powder, 900 times of solution irrigate sterilization, and humidity remains between 70%-85%, avoids direct sunlight, light transmittance 50%-60%.
E) underplanting
Planting time late Febuary-early June, early September-December.Growing the best time is September-November.Through practice, in the underwood planting of height above sea level 500-800 rice, be 99% in cruel summer survival rate.
Rational close planting is important measures of roxburgh anoectochilus terminal bud high yield, ground fertilizer should be dredged, thin should be close, seeding row spacing 5*5 centimetre.Should not plant too dark, the cultivation height of bed 25 centimetres, bed surface is wide 1.2 meters, density 800 strains/m
2,size divides cultivation, be convenient to management and harvest grade distinguishing, compacting seedling surrounding after plantation, water once permeable with mountain spring water after plantation, reach soil completely moistening, apply fertilizer the roxburgh anoectochilus terminal bud season of growth every two weeks use farmers''s liquid fertilizer or gas-liquid, be diluted to 1000 times to carry out, must guard against during fertilising and pollute roxburgh anoectochilus terminal bud blade, if any careless pollution, clear water must be sprayed immediately.Underwood planting situation is shown in Fig. 6.
F) management after transplanting
Keep soil temperature 13 DEG C, temperature 20-25 DEG C; Cultivating 5 days plant can survive, is 99% by the survival rate of underwood planting roxburgh anoectochilus terminal bud.But it is excessive to note watering, rather dryly not wet.
G) extermination of disease and insect pest
Due to sylvan life ecotope more complicated, add that roxburgh anoectochilus terminal bud is the food that rat snake bird winged insect is liked gnawing, so must fence up with object net after plantation.Snail, two spotted spider mite, slug are many, can manual catching and killing, and the diseases such as stem wilt, damping off, damping off, pull out in time or execute organic agricultural chemicals.
H) sylvan life is gathered
Determination and the planting time length of Harvesting Date have substantial connection, must cultivate more than 6 months plant heights more than 10 centimetres, can gather, see Fig. 7 when growing 6 leaves in sylvan life nature.At this moment roxburgh anoectochilus terminal bud internal substance abundant roxburgh anoectochilus terminal bud plant is healthy, stem is sturdy, and well developed root system, aerial root are obvious, and leaf color is deep, flannelette shape is obvious, vein gold thread is distinct, survival rate 99%, and every strain weight reaches 3.5-5.5 gram, pluck to adopt manually to pull out and pluck, advocating with pulling out with putting, gently pulling out and putting down gently.
I) clean
Put well according to magnitude classification when dialling seedling, fresh plant carries out clearing up by strain, remove the plant surface earth especially soil of root, impurity and part epizoite, current are avoided directly to wash away blade, affect roxburgh anoectochilus terminal bud product phase, the roxburgh anoectochilus terminal bud through underwood planting increases by 2 times than the weight of bottle transplantation of seedlings phase in output.
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can make fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dry or dry product three kinds, sees Fig. 8, Fig. 9.
Roxburgh anoectochilus terminal bud adopts rear application: fresh roxburgh anoectochilus terminal bud directly enters meat and fish dishes, integration of drinking and medicinal herbs, is medicine and food.Gold thread lotus leaf grinds is entered meat and fish dishes, mainly cooks the dish of some soup custards.Select fresh roxburgh anoectochilus terminal bud to clean up to put juice extractor to carry out fresh squeezing roxburgh anoectochilus terminal bud juice, directly drink after adding first-class honey and have good effect for the pyrolysis heat that disappears, anti-inflammatory, raising immunity.Adopt the preservation by low temperature technology of freeze-drying (subzero 30 ° of C), make roxburgh anoectochilus terminal bud fresh-keeping more lasting.
embodiment 2
1) draw materials: wild roxburgh anoectochilus terminal bud plants stems section, pick up from Yongtai, Fujian height above sea level 300m mountain stream, wrap with wet cloth after plucking and take back;
2) materials disinfection process: by clean for the wild roxburgh anoectochilus terminal bud tap water gathered, in distance bud point 1.5cm place clip stem section, remove unnecessary blade, put rinsing 15min in saturated bleaching powder supernatant and scrub gently with banister brush, after rinsing well, being placed in flowing water punching and dripping 1 hour, 2 times are rinsed again with distilled water, with the alcohol disinfecting 30s of 75% in superclean bench, add 0.1% mercuric chloride process 10min, then it is for subsequent use to rush 4 times rearmounted workbench with sterile water;
3) adventitious bud induction culture: the material after sterilization is placed on sterilized culture dish, the cut out portion 0.3cm contacted with mercuric chloride is cut away with scalpel, then material is cut into the long stem section of 1.5cm and is inoculated in high temperature, autoclaved adventitious bud induction culture base that (culture medium prescription is with embodiment 1 respectively, medium thickness 1.4cm), cultivation temperature is 21 DEG C, light application time 10h/d, cultivate and carry out black dim cultivation in first 10 days, after 11 days, intensity of illumination is adjusted to 1000lx, bud inducement cultivates 35 days, obtains Multiple Buds;
4) one-step-seedling formation is cultivated: will obtain Multiple Buds through Fiber differentiation, high 4cm, tool 4 internode, the unrooted seedling of 2 leaves, the stem section being cut into 1cm long is inoculated in high temperature, autoclaved one-step-seedling formation medium, and the formula of medium is 1/3MS+N6-isopentene base adenine 1.5mgL
-1+ NAA 2mgL
-1+ HyponexI 3gL
-1+ Hyponex2 3gL
-1+ Peptone 3gL
-1+ potato 150gL
-1+ sucrose 35gL
-1+ active carbon 2gL
-1+ agar 6.5gL
-1, medium thickness is 1.4cm.Stem section must tile storing, and cultivation temperature is 21 DEG C, light application time 12h/d, and intensity of illumination is 1500lx, cultivate within 5 months, obtain the complete and rooting rate of growth reach 99% can directly transplanting bottle seedling plant;
5) bottle transplantation of seedlings: synchronously will to take root and plant height is the roxburgh anoectochilus terminal bud bottle seedling of 10cm through one-step-seedling formation Fiber differentiation 5 months, be placed on natural daylight lower refining seedling 4 days, open bottle cap hardening again 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: comprise select with planting, clear up with planting, put up shade, matrix is disinfected, underplanting, manage after transplanting, sylvan life gathers.Key be to select that height above sea level 300 meters of soil properties are loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, before transplanting, matrix 45% carbendazol wettable powder, 900 times of solution irrigate sterilization; During plantation, every seeding row spacing 5*5cm, cultivation height of bed 30cm, the wide 1.2m of bed surface, density 1000 strains/m
2; During cultivation, keep ground moistening and temperature 15 DEG C, apply fertilizer the roxburgh anoectochilus terminal bud season of growth every two weeks use farmers''s liquid fertilizer or gas-liquid, be diluted to 1000 times of uses; Cultivate more than 7 months more than plant height 12cm in sylvan life nature, can gather when growing 5 leaves.The every strain weight of the roxburgh anoectochilus terminal bud of gathering reaches 3.5 ~ 5.5 grams, is 99% by the survival rate of underwood planting roxburgh anoectochilus terminal bud;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can make fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dry or dry product three kinds.
embodiment 3
1) draw materials: wild roxburgh anoectochilus terminal bud plants stems section, pick up from Yongtai, Fujian height above sea level 600m mountain stream, wrap with wet cloth after plucking and take back;
2) materials disinfection process: by clean for the wild roxburgh anoectochilus terminal bud tap water gathered, in distance bud point 1cm place clip stem section, remove unnecessary blade, put rinsing 15min in saturated bleaching powder supernatant and scrub gently with banister brush, after rinsing well, being placed in flowing water punching and dripping 0.5 hour, 3 times are rinsed again with distilled water, with the alcohol disinfecting 30s of 75% in superclean bench, add 0.1% mercuric chloride process 5min, then it is for subsequent use to rush 3 times rearmounted workbench with sterile water;
3) adventitious bud induction culture: the material after sterilization is placed on sterilized culture dish, the cut out portion 0.4cm contacted with mercuric chloride is cut away with scalpel, then material is cut into the long stem section of 1cm and is inoculated in high temperature, autoclaved adventitious bud induction culture base that (culture medium prescription is with embodiment 1 respectively, medium thickness 1.6cm), cultivation temperature is 25 DEG C, light application time 10h/d, cultivate and carry out black dim cultivation in first 10 days, after 11 days, intensity of illumination is adjusted to 500lx, bud inducement cultivates 45 days, obtains Multiple Buds;
4) one-step-seedling formation is cultivated: will obtain Multiple Buds through Fiber differentiation, high 3.5cm, tool 3 internode, the unrooted seedling of 1 leaf, the stem section being cut into 1.5cm long is inoculated in high temperature, autoclaved one-step-seedling formation medium, and the formula of medium is 1/3MS+N6-isopentene base adenine 0.5mgL
-1+ NAA 1mgL
-1+ HyponexI 2gL
-1+ Hyponex2 2gL
-1+ Peptone 1gL
-1+ potato 100gL
-1+ sucrose 35gL
-1+ active carbon 1gL
-1+ agar 6.5gL
-1, medium thickness is 1.6cm.Stem section must tile storing, and cultivation temperature is 25 DEG C, light application time 12h/d, and intensity of illumination is 2500lx, cultivate within 4 months, obtain the complete and rooting rate of growth reach 99% can directly transplanting bottle seedling plant;
5) bottle transplantation of seedlings: synchronously will to take root and plant height is the roxburgh anoectochilus terminal bud bottle seedling of 6cm through one-step-seedling formation Fiber differentiation 4 months, be placed on natural daylight lower refining seedling 3 days, open bottle cap hardening again 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: comprise select with planting, clear up with planting, put up shade, matrix is disinfected, underplanting, manage after transplanting, sylvan life gathers.Key be to select that height above sea level 800 meters of soil properties are loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, before transplanting, matrix 45% carbendazol wettable powder, 900 times of solution irrigate sterilization; During plantation, every seeding row spacing 5*5cm, cultivation height of bed 20cm, the wide 1m of bed surface, density 500 strains/m
2; During cultivation, keep ground moistening and temperature 12 DEG C, apply fertilizer the roxburgh anoectochilus terminal bud season of growth every two weeks use farmers''s liquid fertilizer or gas-liquid, be diluted to 1000 times of uses; Cultivate more than 8 months more than plant height 15cm in sylvan life nature, can gather when growing 8 leaves.The every strain weight of the roxburgh anoectochilus terminal bud of gathering reaches 3.5 ~ 5.5 grams, is 99% by the survival rate of underwood planting roxburgh anoectochilus terminal bud;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can make fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dry or dry product three kinds.
Claims (1)
1. Wild in Fujian Province roxburgh anoectochilus terminal bud field naturally returns and moves a ground implantation methods, it is characterized in that: concrete steps are as follows:
1) draw materials: wild roxburgh anoectochilus terminal bud plants stems section, pick up from Yongtai, Fujian height above sea level 300 ~ 600m mountain stream, wrap with wet cloth after plucking and take back;
2) materials disinfection process: by clean for the wild roxburgh anoectochilus terminal bud tap water gathered, in distance bud point 1 ~ 2cm place clip stem section, remove unnecessary blade, put rinsing 15min in saturated bleaching powder supernatant and scrub gently with banister brush, after rinsing well, being placed in flowing water punching and dripping 0.5 ~ 1 hour, rinse 2 ~ 3 times with distilled water again, with the alcohol disinfecting 30s of 75% in superclean bench, add 0.1% mercuric chloride process 5 ~ 10min, more for subsequent use with sterile water punching 3 ~ 4 times rearmounted workbench;
3) adventitious bud induction culture: the material after sterile-processed is placed on sterilized culture dish, cut out portion 0.2 ~ the 0.4cm contacted with mercuric chloride is cut away with scalpel, then stem section material being cut into 1 ~ 2cm long is inoculated in respectively in high temperature, autoclaved adventitious bud induction culture base, cultivation temperature is 23 ± 2 DEG C, light application time 10h/d, cultivates and carries out dark culturing in first 10 days, and after 11 days, intensity of illumination is adjusted to 500 ~ 1000lx, bud inducement is cultivated 35 ~ 45 days, obtains Multiple Buds; Wherein, the formula of adventitious bud induction culture base is MS+BA 2mgL
-1+ NAA 1mgL
-1+ KT 0.5mgL
-1+ N6-isopentene base adenine 1mgL
-1+ sucrose 30gL
-1+ active carbon 1gL
-1+ agar 6.5gL
-1, medium thickness is 1.4 ~ 1.6cm;
4) one-step-seedling formation is cultivated: will cultivate the Multiple Buds obtained through bud inducement, high 3 ~ 4cm, tool 3 ~ 4 internode, the unrooted seedling of 1 ~ 2 leaf, the stem section being cut into 1 ~ 2cm long is inoculated in high temperature, autoclaved one-step-seedling formation medium, and stem section must tile storing, cultivation temperature is 23 ± 2 DEG C, light application time 12h/d, intensity of illumination is 1500 ~ 2500lx, cultivates and within 4 ~ 5 months, obtains the bottle seedling plant that the complete and rooting rate of growth reaches 99%; Wherein, the formula of one-step-seedling formation medium is 1/3MS+N6-isopentene base adenine 0.5 ~ 1.5mgL
-1+ NAA 1 ~ 2mgL
-1+ HyponexI 2 ~ 3gL
-1+ Hyponex2 2 ~ 3gL
-1+ Peptone 1 ~ 3gL
-1+ potato 100 ~ 150gL
-1+ sucrose 35gL
-1+ active carbon 1 ~ 2gL
-1+ agar 6.5gL
-1, medium thickness is 1.4 ~ 1.6cm;
5) bottle transplantation of seedlings: synchronously will to take root and plant height is the roxburgh anoectochilus terminal bud bottle seedling of 6 ~ 10cm through one-step-seedling formation Fiber differentiation 4 ~ 5 months, be placed on natural daylight lower refining seedling 3 ~ 5 days, open bottle cap hardening again 2 ~ 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: select that height above sea level 300 ~ 800m soil property is loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, transplant front matrix 45% carbendazol wettable powder, 900 times of solution and irrigate sterilization; During plantation, every seeding row spacing 5*5cm, the cultivation height of bed 20 ~ 30cm, the wide 1 ~ 1.2m of bed surface, density 500 ~ 1000 strains/m
2; During cultivation, keep ground moistening and temperature 12 ~ 15 DEG C, applying fertilizer uses farmers''s liquid fertilizer in the roxburgh anoectochilus terminal bud season of growth every two weeks, is diluted to 1000 times of uses; At sylvan life nature cultivation 6 ~ 8 months plant height 10 ~ 15cm, gathering when growing 5 ~ 8 leaves, is 99% by the survival rate of underwood planting roxburgh anoectochilus terminal bud;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering makes fresh roxburgh anoectochilus terminal bud product respectively, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dry or dry product three kinds.
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| CN103960030A (en) * | 2014-05-23 | 2014-08-06 | 福建省邵武市二都国有林场 | Method for cultivating anoectochilus formosanus in wild imitation mode |
| CN105766647B (en) * | 2016-04-05 | 2018-01-12 | 福建农林大学 | A kind of hectolitre pine method for tissue culture |
| CN106508356A (en) * | 2016-11-01 | 2017-03-22 | 峨眉山生物资源实验站 | Underwood planting method of Emei Anoectochilus formosanus |
| CN106613172B (en) * | 2016-11-28 | 2020-03-10 | 广州甘润堂生物科技有限公司 | Original ecological planting method of anoectochilus roxburghii in Guangdong-oriented chemical-change lake sky and land |
| CN109937877A (en) * | 2019-03-21 | 2019-06-28 | 漳州职业技术学院 | A kind of breeding method of organic selenium-rich roxburgh anoectochilus terminal bud |
| CN110959530A (en) * | 2019-12-12 | 2020-04-07 | 中国林业科学研究院亚热带林业研究所 | Non-distance calanthe field regression method |
| CN111213586B (en) * | 2020-02-26 | 2022-09-13 | 四川农业大学 | Anoectochilus roxburghii tissue culture medium and preparation method thereof |
| CN112931008B (en) * | 2021-02-09 | 2022-12-20 | 中国科学院昆明植物研究所 | Regression planting method for national spice plant hemp roots |
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