Summary of the invention
The object of the invention one is to provide the disposable seedling culture technique of a kind of application, moves ground growing and cultivation roxburgh anoectochilus terminal bud for sylvan life and promotes that industry development provides key technology.The 2nd, solve the roxburgh anoectochilus terminal bud seed small, embryonic development is incomplete, the difficult problem of germinateing under field conditions (factors); Solve interior contained strong analgesia, anti-inflammatory, the anticancer component of roxburgh anoectochilus terminal bud medicine source rareness, particularly plant in prior art and 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 returns naturally moves the ground planting technology, it is characterized in that comprising draw materials, materials disinfection processing, adventitious bud induction culture, disposable seedling are cultivated, bottle transplantation of seedlings, underwood planting, finished product preparation.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 and take back with wet cloth after plucking;
2) materials disinfection is processed: the wild roxburgh anoectochilus terminal bud gathered is rinsed well with running water, in distance bud point 1 ~ 2cm place clip stem section, remove unnecessary blade, put in saturated bleaching powder supernatant rinsing 15min and scrub gently with banister brush, after rinsing well, being placed in the flowing water punching and dripping 0.5 ~ 1 hour, with distilled water, rinse 2 ~ 3 times again, in superclean bench, with 75% alcohol disinfecting 30s, add 0.1% mercuric chloride to process 5 ~ 10min, more standby with 3 ~ 4 times rearmounted workbench of sterile water punching;
3) adventitious bud induction culture: the material after sterilization is placed on sterilized culture dish, cut away the cut out portion 0.2 ~ 0.4cm contacted with mercuric chloride with scalpel, then material being cut into to the long stem section of 1 ~ 2cm is inoculated in respectively in high temperature, autoclaved adventitious bud induction culture base, cultivation temperature is 23 ± 2 ℃, light application time 10h/d, cultivate and within first 10 days, deceive dim cultivation, and after 11 days, intensity of illumination is adjusted into 500-1000lx, bud is induced and is cultivated 35 ~ 45 days, obtains Multiple Buds;
4) disposable seedling is cultivated: will obtain Multiple Buds through inducing to cultivate, high 3 ~ 4cm, tool 3 ~ 4 internodes, the unrooted seedling of 1 ~ 2 leaf, be cut into the long stem section of 1 ~ 2cm and be inoculated in high temperature, autoclaved disposable seedling medium, the storing of must tiling of stem section, cultivation temperature is 23 ± 2 ℃, light application time 12h/d, intensity of illumination is 1500~2500lx, but cultivate, within 4 ~ 5 months, obtains the complete and rooting rate of growth and reaches 99% directly transplanting bottle seedling plant;
5) bottle transplantation of seedlings: will induce the roxburgh anoectochilus terminal bud bottle seedling that cultivation is also synchronously taken root in 4 ~ 5 months and plant height is 6 ~ 10cm through disposable seedling, be placed on the natural daylight lower refining seedling 3 ~ 5 days, open again the bottle cap hardening 2 ~ 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: select that 300 ~ 800 meters soil properties of height above sea level are loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, transplant front matrix and irrigate sterilization with 900 times of solution of 45% carbendazol wettable powder; 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 12 ~ 15 ℃ of ground moistening and temperature, fertilising 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 while growing 5 ~ 8 leaves, and the survival rate by the underwood planting roxburgh anoectochilus terminal bud is 99%;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can be made fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dries or dry three kinds of products.
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 the disposable seedling medium of described step 4) 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 disposable seedling technology groups training roxburgh anoectochilus terminal bud, improved roxburgh anoectochilus terminal bud transplanted seedling survival rate and rooting rate.Naturally return plantation by sylvan life with moving, discharge the content of some hormones of incubation, the accumulation that further improves growing ability and the benefit materials of roxburgh anoectochilus terminal bud, after plantation, roxburgh anoectochilus terminal bud increases by 1 ~ 2 times than bottle transplantation of seedlings phase weight.
2, the present invention sets up disposable seedling culture technique, transplanting and simulation sylvan life growth; naturally return a complete set of production technology system of plantation with moving; 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; accelerated the capital fund collection process; meet the market demand, saved investment enterprise's plenty of time and fund.
3, economize the land resource; the protection peasant benefit; in agricultural structure is adjusted; take full advantage of forest land resource; development is applicable to the Wild in Fujian Province roxburgh anoectochilus terminal bud of sylvan life growth, and peasant's vested interest is without prejudice like this, does not only affect the agricultural production while but also has created the agricultural products of high added value; increase farmers' income, truly accomplished and reached 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 the forest land utilization rate, greatly improve the economic benefit of forest, for mountain area labour's reasonable arrangement provides condition, simultaneously for returning and the labour in abundant current mountain area is best appeal.
5, be conducive to the forest ecology protection, in conjunction with fostering, play branch, topdressing, promote tree growth.
6, sylvan life is carried out artificial cultivation and can be met the required growing environment of roxburgh anoectochilus terminal bud wild state, the input of simultaneously saving sunshade net.Therefore, protection roxburgh anoectochilus terminal bud these species have important scientific research, medical science, ecological significance and value, will play an important role for mankind's treating disease and health.Calculated and be take 6 months as a production cycle according to current plantation situation, develop at a progressive speed, one mu of annual plantation in mountain region can have twice harvest, 300 yuan/kg of fresh goods by (current market price) underwood planting, every mu of results fresh goods 500kg*2, every mu of annual value of production 600000 RMB, so the Anoectochilus roxburghii underwood planting has a extensive future.
The present invention bases oneself upon Fujian mountain area forest land resource advantage, " carry out the intercropping of woods medicine, ecological cultivation " principle, to roxburgh anoectochilus terminal bud, adopt sylvan life to return cultivation with moving, build the optimum growh environment that is applicable to roxburgh anoectochilus terminal bud bottle seedling, improve transplanting success, form the scale of roxburgh anoectochilus terminal bud group training seedling underwood planting, set up cultivation system under the forest plantation of Anoectochilus roxburghii, improving roxburgh anoectochilus terminal bud Industry Promotion system so neither takies the peasant and can plough soil, can preserve the ecological environment again, through for many years, the multiple spot experiment in cultivation, development Anoectochilus roxburghii industry, must carry out the sylvan life exploitation, implement ecological cultivation, reach the effect of economic benefit and ecological benefits doulbe-sides' victory.
Embodiment
For fully disclose Wild in Fujian Province roxburgh anoectochilus terminal bud of the present invention field naturally return move planting technology, below the present invention is described further, but the invention is not restricted to this.
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 and take back with wet cloth after plucking.
2) materials disinfection is processed: the wild roxburgh anoectochilus terminal bud gathered is rinsed well with running water, in distance bud point 2cm place clip stem section, remove unnecessary blade, put in saturated bleaching powder supernatant rinsing 15min and scrub gently with banister brush, after rinsing well, being placed in the flowing water punching and dripping 1 hour, with distilled water, rinse 3 times again, in superclean bench, with 75% alcohol disinfecting 30s, add 0.1% mercuric chloride to process 8min, more standby with 4 times rearmounted workbench of sterile water punching.
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, then material is cut into to the long stem section of 2cm and 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 ℃, and light application time 10h/d cultivates and within first 10 days, deceives dim cultivation, and after 11 days, intensity of illumination is adjusted into 800lx, and bud is induced and cultivated 40 days, obtains Multiple Buds.
4) disposable seedling is cultivated: will induce to cultivate through bud to obtain Multiple Buds, and high 3cm, tool 3 internodes, the unrooted seedling of 2 leaves, be cut into the long stem section of 2cm and be inoculated in high temperature, autoclaved disposable seedling medium, and the storing of must tiling of stem section, be shown in 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 ℃, light application time 12h/d, and intensity of illumination is 2000lx, cultivates within 4.5 months, to obtain the complete and rooting rate of growth and reach 99% and can directly transplant a bottle seedling plant, sees Fig. 2, Fig. 3.
5) bottle transplantation of seedlings: will induce the roxburgh anoectochilus terminal bud bottle seedling that cultivation is also synchronously taken root in 4.5 months and plant height is 8cm through disposable seedling, and be placed on the natural daylight lower refining seedling 5 days, then open the bottle cap hardening 2 days, and see Fig. 4.Then take out plant from blake bottle, wash away the medium be attached on root system.
6) underwood planting: comprise with planting select, clear up with planting, put up that shade, matrix are disinfected, manage after underplanting, transplanting, sylvan life gathers.
A) select with planting
Select that 500 meters soil properties of height above sea level 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%, suitable life temperature is 15-28 ℃, roxburgh anoectochilus terminal bud should be grown under the Acid soil condition.
B) cleaning with planting
Land readjustment, weed with a hoe, to around shrub and some tree branches dredge between carrying out, surgery, make simultaneously flat, the wide 30-60 centimetre in the ridge, high 20 centimetres, also can plant according to mountain region actual that position arranges and the tree line-spacing is grasped flexibly.Under field conditions (factors), upper strata arbor crown density 65%-75%, low cover cover degree 35-45%.
C) put up shade
Put up shade, regulate light transmittance, printing opacity be take three sun seven the moon as being applicable to, and 4 ~ May, temperature raise, and printing opacity can be reduced to below 30%, must adopt the fences basketry around planting site, draw and cover the invasion and attack that sunshade net prevents the wildlife harm such as mouse, snake, bird, worm.
Fig. 5 is seen in the underwood planting habitat.
D) matrix is disinfected
Before transplanting, matrix irrigates sterilization with 900 times of solution of 45% carbendazol wettable powder, and humidity remains between 70%-85%, avoids direct sunlight, light transmittance 50%-60%.
E) underplanting
Planting time late Febuary-early June, early September-December.Best time of growing is September-November.Through practice, in the underwood planting of height above sea level 500-800 rice, in cruel summer survival rate, be 99%.
Rational close planting is important measures of roxburgh anoectochilus terminal bud high yield, ground fertilizer should dredge, thin should be close, seeding row spacing 5*5 centimetre.Should not plant too dark, 25 centimetres of the height of bed of cultivation, bed surface is wide 1.2 meters, density 800 strains/m
2,size minute is planted, be convenient to management and harvest grade distinguishing, compacting seedling surrounding after plantation, water once permeablely after plantation with mountain spring water, reach soil fully moistening, fertilising the roxburgh anoectochilus terminal bud season of growth every two weeks use farmers''s liquid fertilizer or gas-liquid, being diluted to 1000 times carries out, must guard against during fertilising and pollute the roxburgh anoectochilus terminal bud blade, if any careless pollution, must spray immediately clear water.The underwood planting situation is shown in Fig. 6.
F) transplant rear management
Keep 13 ℃ of soil temperature, temperature 20-25 ℃; Cultivating 5 days plant can survive, and the survival rate by the underwood planting roxburgh anoectochilus terminal bud is 99%.But it is not it is excessive to note watering, peaceful dry wet.
G) extermination of disease and insect pest
Due to sylvan life ecotope more complicated, add that roxburgh anoectochilus terminal bud is that rat snake bird winged insect is liked the food of gnawing, so must fence up with the purpose net after plantation.Snail, two spotted spider mite, slug are many, can the manual catching and killing, and the diseases such as stem wilt, damping off, damping off, pull out or execute organic agricultural chemicals in time.
H) sylvan life is gathered
Determining with planting time length of Harvesting Date has substantial connection, must cultivate plant height more than 6 months in the sylvan life nature and, more than 10 centimetres, can gather while growing 6 leaves, sees Fig. 7.At this moment the abundant roxburgh anoectochilus terminal bud plant of roxburgh anoectochilus terminal bud internal substance is healthy, stem is sturdy, and well developed root system, aerial root are obvious, and leaf color is deep, the flannelette shape is obvious, the vein gold thread is distinct, and survival rate reaches 99%, and every strain weight reaches the 3.5-5.5 gram, pluck to adopt and manually pull out and pluck, advocate with pulling out with putting, gently pull out and put down gently.
I) clean
While dialling seedling, according to magnitude classification, put well, bright plant pursues the strain cleaning, remove plant surface earth especially soil, impurity and the part epizoite of root, avoid current directly to wash away blade, affect roxburgh anoectochilus terminal bud product phase, through the roxburgh anoectochilus terminal bud of underwood planting, on output, than the weight of bottle transplantation of seedlings phase, increase by 2 times.
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can be made fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dries or dry three kinds of products, sees Fig. 8, Fig. 9.
Roxburgh anoectochilus terminal bud is adopted rear application: fresh roxburgh anoectochilus terminal bud is directly into meat and fish dishes, and integration of drinking and medicinal herbs, be medicine and food.Gold thread lotus leaf grinds is entered to meat and fish dishes, is mainly the dish of doing some soup custards.Select fresh roxburgh anoectochilus terminal bud to clean up to put juice extractor to carry out aquatic foods and press roxburgh anoectochilus terminal bud juice, adding after first-class honey directly to drink for the pyrolysis heat that disappears, anti-inflammatory, raising immunity has good effect.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 and take back with wet cloth after plucking;
2) materials disinfection is processed: the wild roxburgh anoectochilus terminal bud gathered is rinsed well with running water, in distance bud point 1.5cm place clip stem section, remove unnecessary blade, put in saturated bleaching powder supernatant rinsing 15min and scrub gently with banister brush, after rinsing well, being placed in the flowing water punching and dripping 1 hour, with distilled water, rinse 2 times again, in superclean bench, with 75% alcohol disinfecting 30s, add 0.1% mercuric chloride to process 10min, more standby with 4 times rearmounted workbench of sterile water punching;
3) adventitious bud induction culture: the material after sterilization is placed on sterilized culture dish, cut away the cut out portion 0.3cm contacted with mercuric chloride with scalpel, then material is cut into to the long stem section of 1.5cm and is inoculated in respectively that (culture medium prescription is with embodiment 1 in high temperature, autoclaved adventitious bud induction culture base, medium thickness 1.4cm), cultivation temperature is 21 ℃, light application time 10h/d, cultivate and within first 10 days, deceive dim cultivation, after 11 days, intensity of illumination is adjusted into 1000lx, bud is induced and is cultivated 35 days, obtains Multiple Buds;
4) disposable seedling is cultivated: will obtain Multiple Buds through inducing to cultivate, high 4cm, tool 4 internodes, the unrooted seedling of 2 leaves, be cut into the long stem section of 1cm and be inoculated in high temperature, autoclaved disposable seedling medium, 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.The storing of must tiling of stem section, cultivation temperature is 21 ℃, light application time 12h/d, intensity of illumination is 1500lx, but cultivate, within 5 months, obtains the complete and rooting rate of growth and reaches 99% directly transplanting bottle seedling plant;
5) bottle transplantation of seedlings: will induce the roxburgh anoectochilus terminal bud bottle seedling that cultivation is also synchronously taken root in 5 months and plant height is 10cm through disposable seedling, be placed on the natural daylight lower refining seedling 4 days, open again the bottle cap hardening 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: comprise with planting select, clear up with planting, put up that shade, matrix are disinfected, manage after underplanting, transplanting, sylvan life gathers.Key be to select that 300 meters soil properties of height above sea level are loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, before transplanting, matrix irrigates sterilization with 900 times of solution of 45% carbendazol wettable powder; 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 15 ℃ of ground moistening and temperature, fertilising 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; In sylvan life nature cultivation, more than 7 months more than plant height 12cm, while growing 5 leaves, can gather.The every strain weight of the roxburgh anoectochilus terminal bud of gathering reaches 3.5 ~ 5.5 grams, and the survival rate by the underwood planting roxburgh anoectochilus terminal bud is 99%;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can be made fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dries or dry three kinds of products.
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 and take back with wet cloth after plucking;
2) materials disinfection is processed: the wild roxburgh anoectochilus terminal bud gathered is rinsed well with running water, in distance bud point 1cm place clip stem section, remove unnecessary blade, put in saturated bleaching powder supernatant rinsing 15min and scrub gently with banister brush, after rinsing well, being placed in the flowing water punching and dripping 0.5 hour, with distilled water, rinse 3 times again, in superclean bench, with 75% alcohol disinfecting 30s, add 0.1% mercuric chloride to process 5min, more standby with 3 times rearmounted workbench of sterile water punching;
3) adventitious bud induction culture: the material after sterilization is placed on sterilized culture dish, cut away the cut out portion 0.4cm contacted with mercuric chloride with scalpel, then material is cut into to the long stem section of 1cm and is inoculated in respectively that (culture medium prescription is with embodiment 1 in high temperature, autoclaved adventitious bud induction culture base, medium thickness 1.6cm), cultivation temperature is 25 ℃, light application time 10h/d, cultivate and within first 10 days, deceive dim cultivation, after 11 days, intensity of illumination is adjusted into 500lx, bud is induced and is cultivated 45 days, obtains Multiple Buds;
4) disposable seedling is cultivated: will obtain Multiple Buds through inducing to cultivate, high 3.5cm, tool 3 internodes, the unrooted seedling of 1 leaf, be cut into the long stem section of 1.5cm and be inoculated in high temperature, autoclaved disposable seedling medium, 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.The storing of must tiling of stem section, cultivation temperature is 25 ℃, light application time 12h/d, intensity of illumination is 2500lx, but cultivate, within 4 months, obtains the complete and rooting rate of growth and reaches 99% directly transplanting bottle seedling plant;
5) bottle transplantation of seedlings: will induce the roxburgh anoectochilus terminal bud bottle seedling that cultivation is also synchronously taken root in 4 months and plant height is 6cm through disposable seedling, be placed on the natural daylight lower refining seedling 3 days, open again the bottle cap hardening 3 days, then from blake bottle, take out plant, wash away the medium be attached on root system;
6) underwood planting: comprise with planting select, clear up with planting, put up that shade, matrix are disinfected, manage after underplanting, transplanting, sylvan life gathers.Key be to select that 800 meters soil properties of height above sea level are loose, ventilative, draining, mountain region that water retention property is good, avoid direct sunlight, before transplanting, matrix irrigates sterilization with 900 times of solution of 45% carbendazol wettable powder; 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 12 ℃ of ground moistening and temperature, fertilising 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; In sylvan life nature cultivation, more than 8 months more than plant height 15cm, while growing 8 leaves, can gather.The every strain weight of the roxburgh anoectochilus terminal bud of gathering reaches 3.5 ~ 5.5 grams, and the survival rate by the underwood planting roxburgh anoectochilus terminal bud is 99%;
7) finished product preparation: the roxburgh anoectochilus terminal bud after gathering can be made fresh roxburgh anoectochilus terminal bud product, the freeze-drying prods of roxburgh anoectochilus terminal bud, roxburgh anoectochilus terminal bud dries or dry three kinds of products.