CN106613172A - Method for original ecological planting of genuine shorthairy antenoron in Guangdong Conghua Heavenly lake - Google Patents

Method for original ecological planting of genuine shorthairy antenoron in Guangdong Conghua Heavenly lake Download PDF

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Publication number
CN106613172A
CN106613172A CN201611064121.8A CN201611064121A CN106613172A CN 106613172 A CN106613172 A CN 106613172A CN 201611064121 A CN201611064121 A CN 201611064121A CN 106613172 A CN106613172 A CN 106613172A
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China
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terminal bud
roxburgh anoectochilus
anoectochilus terminal
lake
day
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CN106613172B (en
Inventor
吴坤林
叶志聪
张新华
李鉴深
段俊
吴志斌
张彦
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Guangzhou Ganruntang Biological Technology Co ltd
South China Botanical Garden of CAS
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Guangzhou Ganruntang Biological Technology Co ltd
South China Botanical Garden of CAS
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00Cultivation of specific crops or plants not otherwise provided for
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001Culture apparatus for tissue culture
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008Methods for regeneration to complete plants

Abstract

The invention discloses a method for original ecological planting of genuine shorthairy antenoron in Guangdong Conghua Heavenly lake. The method comprises the following steps that (A) Heavenly lake shorthairy antenoron tissue culture seedlings are obtained; (B) forest land selection and preparation are performed; (C) the Heavenly lake shorthairy antenoron tissue culture seedlings are planted; (D) field management is performed; (E) harvesting is performed, wherein the Heavenly lake shorthairy antenoron is harvested in harvesting season. The method successfully utilizes seeds high in germination rate, a culture medium rich in nutrition and a full-natural-light culture room to perform one-time and large-scale production of shorthairy antenoron seedlings, has the advantages of being low in cost and high in efficiency, is a high and new biotechnology for large-scale seedling production and can promote sustainable development of the shorthairy antenoron industry. The method for original ecological planting of the genuine shorthairy antenoron in Guangdong Conghua Heavenly lake ensures the original performance and treating effect of the variety, has the advantages of being low in cost, strong in genuineness, high in medicinal effect and high in selling price, is a method for original ecological planting of precious traditional Chinese medicinal materials and is unique, simple, affordable and high in application value.

Description

A kind of genuine roxburgh anoectochilus terminal bud ecosystem implantation methods in Conghua of Guangdong Province day lake
Technical field:
The invention belongs to field of plant reproduction, and in particular to a kind of genuine roxburgh anoectochilus terminal bud ecosystem plantation side in Conghua of Guangdong Province day lake Method.
Background technology:
Roxburgh anoectochilus terminal bud another name Shorthairy Antenoron, rough melic herb, are that orchid family (Orchidaceae) opens lip plant flowers aspidistra category (Anoectochilus Bl.) perennial rare herbaceous plant, China has 23 kinds or so, and which part kind herb is among the people to make medicine With, it is traditional rare rare traditional Chinese medicine, it is wider in use range among the people, is used for treating diabetes, high fat of blood, hepatitis B Etc. disease, the laudatory titles such as " king of medicine ", " gold grass ", " god's medicine ", " black ginseng " are have, be referred to as " king in medicine ".For many years, due to it The market demand is big and cause artificial excessively excavation, add that roxburgh anoectochilus terminal bud natural propagation rate is low, poor growth, wild roxburgh anoectochilus terminal bud resource It is very rare, it is endangered, just roxburgh anoectochilus terminal bud is classified as into one of Chinese medicine of focused protection early in State Council in 1987, it is country Second class protection plant.And because artificial growth roxburgh anoectochilus terminal bud has larger difficulty, the quantity of current artificial growth is also very limited. Due to the restriction of former material doses, in recent years roxburgh anoectochilus terminal bud industry atrophy.Therefore greatly develop artificial growth and seem and very must Will with it is urgent.Because roxburgh anoectochilus terminal bud is extensively applied in clinic, and curative effect is fabulous, with high economic benefit and society's effect Benefit, therefore the market demand is increasing, and the roxburgh anoectochilus terminal bud of artificial growth is also far from the demand for meeting market, existing market price Costliness, the current price per kilogram of the dry product by made by roxburgh anoectochilus terminal bud reaches 10,000 yuan, and market roxburgh anoectochilus terminal bud fresh goods price is also up to 800- 1000 yuan/kilogram, if per mu of energy more than 300 kilograms fresh goods of output, per mu yield value is up to more than 240,000 yuan.It is current agricultural planting One of most effective industry in industry.The artificial growth of roxburgh anoectochilus terminal bud is greatly developed, need of the people to roxburgh anoectochilus terminal bud can be not only met Ask, but also can be because reducing the purpose that protection wild resource is played to the dependence of wild resource, therefore artificial growth gold thread Lotus not only has good market prospects, and the protection to wild resource also has important meaning.
The content of the invention:
It is an object of the invention to provide a kind of have low cost, the genuine roxburgh anoectochilus terminal bud in Conghua of Guangdong Province day lake of efficient feature Ecosystem implantation methods, the day lake roxburgh anoectochilus terminal bud bred by the inventive method has the characteristics of genuineness is strong, and drug effect is high.
The genuine roxburgh anoectochilus terminal bud ecosystem implantation methods in Conghua of Guangdong Province day lake of the present invention, it is characterised in that comprise the following steps:
The acquisition of A, day lake roxburgh anoectochilus terminal bud plantlet in vitro:In the roxburgh anoectochilus terminal bud Proterozoic of Conghua of Guangdong Province day lake, robust growth, stem are chosen Slightly, the day lake roxburgh anoectochilus terminal bud health strain that blade is big, blade reticulate pattern is clear and many, resistance, disease and insect resistance are strong, in day lake roxburgh anoectochilus terminal bud Flowering naturally season carries out artificial pollination and bagging, and fruit grows under natural conditions, and selection fruit age is 50~70 days Capsule is explant, the sterilized moisture for blotting capsule surface with aseptic filter paper under aseptic condition on superclean bench, is connect And cut capsule with aseptic operation knife, broadcast sowing embryo germination and protocorm differentiation growth medium A1, in 23 ± 2 DEG C, illumination 500~1000lux of degree, cultivates under illumination 12h/d, and embryo expands to form seedling of the continued growth formation with root after protocorm, will High 2~3 centimetres of seedling, goes to Rooting and hardening-off culture base A2 cultures, and cultivation temperature is 25 ± 2 DEG C, and illuminance 1000~ 2000lux, illumination 12h/d, obtain day lake roxburgh anoectochilus terminal bud and take root seedling, and day lake roxburgh anoectochilus terminal bud is taken root into seedling in natural light lower refining seedling 7~15 Bottle outlet after it, obtains bottle seedling of taking root, and before plantation, bottle seedling of taking root is taken out from blake bottle, after cleaning the culture medium of attachment, uses quality The millesimal potassium permanganate solution of fraction soaks 5~10min, and a day lake roxburgh anoectochilus terminal bud plantlet in vitro is obtained after draining, and waits to plant;Institute The embryo germination stated and protocorm differentiation growth medium A1, its per liter containing spending treasured No. 1 1~2g, 0.5~2g of peptone, it is active 50~100mg of carbon, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2mg of thiamine hydrochloride, puridoxine hydrochloride 0.4~0.8mg, 0.4~0.8mg of nicotinic acid, 0.2~0.5mg of methyl α-naphthyl acetate, 15~20g of sucrose, 6~7g of agar, balance of water, pH 5.4-5.6;Described Rooting and hardening-off culture base A2, its per liter containing spending No. 1 1~2g of treasured, 0.5~2g of peptone, mashed potatoes 40~ 80g, 500~1000mg of activated carbon, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2mg of thiamine hydrochloride, salt Sour 0.4~0.8mg of pyridoxol, 0.4~0.8mg of nicotinic acid, 0.2~0.5mg of methyl α-naphthyl acetate, 20~30g of sucrose, 6~7g of agar, surplus For water, pH5.4-5.6;
B, forest land select and site preparation:Select day lake roxburgh anoectochilus terminal bud Proterozoic, Subtropical Mountainous of the height above sea level in 100-500m Or hills forest land, vegetation is green throughout the year based on evergreen broadleaf forest and theropencedrymion, well arranged, and canopy density are in 0.85- 0.9, periphery has sufficient pollution-free water source, and long-term air humidity is more than 60%;In nearly small stream limes marginis shady place forest land, forest land is removed Weeds and surface are fallen leaves, and the soil layer of sylvan life is turned over, and ploughed depth is 20-30cm, with the soil layer that ravels, and play kind of a planting-line, The fertile fertile soil that layer overlay periphery hillside fields is collected on furrow;Fallen leaves bearer network is installed on furrow, the blade that trees drop is carried, Avoid covering the day lake roxburgh anoectochilus terminal bud in forest land;Water injection system is installed simultaneously on furrow, can be sprayed water under more dry weather Irrigate, keep the humidity of air;
The plantation of C, day lake roxburgh anoectochilus terminal bud plantlet in vitro:When the annual 4-8 months or other month temperatures being higher than 10 DEG C, by day Lake roxburgh anoectochilus terminal bud plantlet in vitro is implanted on the kind planting-line in forest land, and day lake roxburgh anoectochilus terminal bud plantlet in vitro plantation spacing is 3-5cm, and planting depth is Soil layer hides a day root for lake roxburgh anoectochilus terminal bud, pours root water;
D, forest management:Day lake roxburgh anoectochilus terminal bud kind woodlot relative air humidity is kept to be maintained at using water injection system water spray More than 70%, cleaning fallen leaves ensure that day lake anoectochilus blume seedling can receive the scattered light of abundance, in the roxburgh anoectochilus terminal bud plantlet in vitro growth of day lake Period, water spray process is carried out using water injection system so that plant planting-line and air keeps the state of moistening, apply fertilizer after plant, and prevent Control pest and disease damage;
E, harvesting:Day lake roxburgh anoectochilus terminal bud is harvested to collecting season.
Day lake roxburgh anoectochilus terminal bud plantlet in vitro be colonized 7~9 months can harvesting, now day lake roxburgh anoectochilus terminal bud plant grow to height 6~ More than 10cm, 5~6 leaves, every plant of 2~3g of fresh weight can be harvested.Day lake roxburgh anoectochilus terminal bud wild characters now are good, using effect Really good, drug effect is high, thus its price is higher by more than 1/3rd than the day lake roxburgh anoectochilus terminal bud that Artificial facilities are cultivated.
The sterilization of described step A is preferably soaks explant 30 seconds in the alcohol of volume fraction 70%, then with 0.1% Mercuric chloride solution is sterilized 15~20 minutes, aseptic water washing 4~5 times.
The pouring root water of described step C is preferably after the roxburgh anoectochilus terminal bud plantlet in vitro transplanting of day lake with carbendazim or thiram 1000 times of water dilute solutions are poured as root water.
Apply fertilizer after the plant of described step D, and prevention and elimination of disease and pests preferably transplants one from day lake roxburgh anoectochilus terminal bud plantlet in vitro After month, with solution as Foliar Spraying, spray to the dropping liquid of blade face, later per 7~10 days, fertilising (used solution as blade face Fertilizer is sprayed) once, in disease season occurred frequently, with agricultural streptomycin sulphate 100mg/kg or general 1000 times of water diluents, many are made every effort to overcome The rotations such as bacterium 1000 times of water diluents of spirit are sprayed, and are monthly sprayed 1 time, and snail is born in for 5~June with slug is multiple, is spread with quick lime Impose on surrounding preventing and treating;Described solution be per 15 jin of peanut press pulps plus 150 jin of water spontaneous fermentation 30 days after 8 times of liquid fertilizer dilute with water Afterwards, then spend precious No. 1 foliage fertilizer, 1.7 grams of potassium dihydrogen phosphates of per liter of 2.0 grams of addition is well mixed to form solution.
Easy to operate, low production cost of the invention, is easy to the extensive underwood planting of the genuine roxburgh anoectochilus terminal bud ecosystem in day lake, it is ensured that Its original performance of lake roxburgh anoectochilus terminal bud and curative effect.Using the present invention plantation day lake roxburgh anoectochilus terminal bud, be conducive to keep introduces a collection genuineness and Local Adaptation, promotes the growth of day lake roxburgh anoectochilus terminal bud and the raising of drug effect, improves the use value and safety of the genuine roxburgh anoectochilus terminal bud in day lake Property, there is very active influence to resources of medicinal plant regeneration in short supply in imminent danger and sustainable utilization;Furthermore with the woods of Proterozoic Ground, without the need for building booth and purchase planting matrix, can save many cost payouts, production cost be reduced, while price is compared Higher, economic benefit is more significantly.
Compared to prior art, the technology of the present invention feature is as follows:
1. the key point that the present invention obtains high seed germination rate be to take pollination after the capsule of 50~70 days as explant, when Between too short or long capsule can reduce a day seed germination rate for lake roxburgh anoectochilus terminal bud.
2. it is the growth adaptability of guarantee day lake seedlings of anoectochilus, the day lake roxburgh anoectochilus terminal bud fine individual plant with Proterozoic is as mother This, carries out Proterozoic growth and artificial pollination, and its seed germination rate of the capsule of acquisition of pollinating is high, easy seedling.
3., to ensure day lake roxburgh anoectochilus terminal bud Chinese medicine genuineness, pedestrian is entered using the timbered soil matrix of its original in Proterozoic Work post is planted, and can significantly reduce production cost, and the day lake roxburgh anoectochilus terminal bud produced has wildness, drug effect height, price high Feature.
The many reports of roxburgh anoectochilus terminal bud artificial growth, but be mostly to pluck in explant or field gene bank to enter pedestrian from field Work pollination obtains the artificial propagation that seedling is carried out after seed, then carries out moving ground underwood planting or artificial growth;Generally existing gold The problems such as genuine sex chromosome mosaicism of line lotus Chinese medicine and the not strong Local Adaptation of introduces a collection, cause the effective component of roxburgh anoectochilus terminal bud inadequate Fully affect its using effect.Choosing elite plant strain in roxburgh anoectochilus terminal bud group in the wild of the invention carries out artificial pollination, in primary State is grown and obtains ripe seed, is successfully carried out aseptic seeding using the seed of high germination rate on the spot and is produced roxburgh anoectochilus terminal bud kind Seedling, then seedlings of anoectochilus recurrence ecosystem forest land is carried out into imitating wild planting, with low cost, genuineness is strong, and drug effect is high, price High the characteristics of, be precious Chinese herbal medicine ecosystem implantation methods, and technology is unique and simple material benefit, and using value is high.Implement the invention Carry out by only there need to be simple Plant Tissue Breeding equipment.
The present invention successfully carries out gold using the seed of high germination rate, nutritious culture medium, full natural light culturing room Line lotus seedling once-seedling forming is mass produced, and with low cost, efficient feature is that the height of the large-scale production of seedling is newborn Thing technology, can promote the sustainable development of roxburgh anoectochilus terminal bud industry.It is primary that patent of the present invention provides the genuine roxburgh anoectochilus terminal bud in Conghua of Guangdong Province day lake State implantation methods, it is ensured that the original performance of the kind and curative effect, with low cost, genuineness is strong, and drug effect is high, the high spy of price Point, is precious Chinese herbal medicine ecosystem implantation methods, and technology is unique and simple material benefit, and using value is high.
Seedling large-scale production is carried out in situ using the seed of the high germination rate of Conghua of Guangdong Province day lake roxburgh anoectochilus terminal bud, then will production Seedling out carries out imitating wild planting in ecosystem forest land, it is ensured that the original performance of the kind and curative effect, with low cost, medicine The characteristics of imitating high, genuineness is strong, improves the security that seedlings of anoectochilus are used, to roxburgh anoectochilus terminal bud resources of medicinal plant in short supply in imminent danger again Raw and sustainable utilization is of great significance, and meets great market demand of the health industry to precious Chinese herbal medicine roxburgh anoectochilus terminal bud.
Specific embodiment:
Following examples are that the present invention is further illustrated, rather than limitation of the present invention.
Embodiment 1:
1. Proterozoic artificial pollination:In the roxburgh anoectochilus terminal bud Proterozoic of Conghua of Guangdong Province day lake (37 points to 23 degree 40 points of north latitude 23 degree, 36 points to 113 degree 38 points of east longitude 113 degree), choose robust growth, stem is thick, blade is big, blade reticulate pattern is clear and many, resistance, resist The strong day lake roxburgh anoectochilus terminal bud of pest and disease damage (Anoectochilus roxburghii (wall) lindl. ' tianhu ') health strain, in Its lake roxburgh anoectochilus terminal bud flowering naturally season carries out the information such as artificial pollination and bagging, mark date, and fruit grows under natural conditions Development.
2. aseptic seeding produces seedling:It is explant to choose the day lake roxburgh anoectochilus terminal bud capsule that fruit age is 50~70 days, in volume Soak 30 seconds in the alcohol water blend of fraction 70%, then with the water-soluble liquid disinfectant of the mercuric chloride of mass fraction 0.1% 15~20 minutes, sterilized water Rinse 4~5 times, blot the moisture on capsule surface with aseptic filter paper under aseptic condition on superclean bench.Then aseptic hand is used Art knife cuts capsule, uniform broadcasting to embryo germination and protocorm differentiation growth medium A1 [described embryo germination and protocorm Per liter of stem differential growth culture medium A 1 is to be configured so that:Treasured No. 1 1g, peptone 2g, activated carbon 50mg, inositol 120mg will be spent, it is sweet Propylhomoserin 1.5mg, thiamine hydrochloride (VB1) 0.2mg, puridoxine hydrochloride (VB6) 0.4mg, nicotinic acid 0.8mg, methyl α-naphthyl acetate (NAA) 0.2mg, sucrose 20g, agar 6g, in being added to a small amount of water, are then settled to 1L with water, adjust pH 5.6].After 90 days, plant Embryo expands and to form continued growth after protocorm and form high 2~3 centimetres of seedlings with root.Embryo germination and protocorm differentiation growth step Section cultivation temperature be (23 ± 2) DEG C, 500~1000lux of illuminance, illumination 12h/d.By high 2~3 centimetres of seedling, go to [per liter of Rooting and hardening-off culture base A2 is configured so that Rooting and hardening-off culture base A2:Spend No. 1 1g of treasured, peptone 2g, mashed potatoes 40g, activated carbon 1000mg, inositol 80mg, glycine 2.5mg, thiamine hydrochloride (VB1) 0.05mg, puridoxine hydrochloride (VB6) 0.8mg, nicotinic acid 0.4mg, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 20g, agar 7g, in being added to a small amount of water, are then settled to water 1L, adjusts pH 5.6], during culture 90 days, height of seedling can reach 5~7 centimetres, radical 3~5, and rooting rate is 100%.Strong plantlets and rootage The cultivation temperature in stage be (25 ± 2) DEG C, 1000~2000lux of illuminance, illumination 12h/d.Roxburgh anoectochilus terminal bud seedling of taking root in day lake is existed Natural lighting lower refining seedling bottle outlet after 7~15 days.Before plantation, seedling of taking root is taken out from blake bottle, after cleaning the culture medium of attachment, used The potassium permanganate solution of one thousandth (mass fraction) soaks 5~10min, and a day lake roxburgh anoectochilus terminal bud plantlet in vitro is obtained after draining, and treats Plantation.To go up to the sky, lake seedlings of anoectochilus are produced near to the tissue culture workshop that day lake side is set up.The genuine roxburgh anoectochilus terminal bud in one day lake Capsule by the aseptic seeding culture of 6~August, can quick propagating and cultivating go out more than 100,000 plants of day lake roxburgh anoectochilus terminal bud plantlet in vitro.
3. forest land selects and site preparation:The genuine roxburgh anoectochilus terminal bud Proterozoic in day lake is selected, height above sea level is in the subtropical zone of 100-500m Mountain region or hills forest land, 20.9 degree of year-round average temperature, 38.7 degree of maximum temperature, minimum temperature -2.5 is spent;Average annual rainfall 2100 milliliters, precipitation concentrates the 4-8 months;Vegetation is green throughout the year based on evergreen broadleaf forest and theropencedrymion, well arranged, strongly fragrant In 0.85-0.9, periphery has sufficient pollution-free water source to degree of closing, and long-term air humidity is more than 60%;In nearly small stream limes marginis shady place woods Ground, removes forest land weeds and surface fallen leaves, and the soil layer of sylvan life is turned over, and ploughed depth is 20-30cm, with the soil layer that ravels, And the kind planting-line of 5 meters of wide 60 centimeter length is played into, the fertile fertile soil that layer overlay periphery hillside fields is collected on furrow;Two meter ampere on furrow Dress fallen leaves bearer network, carries the blade that trees drop, it is to avoid cover the day lake roxburgh anoectochilus terminal bud in forest land;Water spray system is installed simultaneously on furrow System, can carry out spray irrigation under more dry weather, keep the humidity of air.
4. the plantation of day lake roxburgh anoectochilus terminal bud plantlet in vitro:Using sterile-processed day lake roxburgh anoectochilus terminal bud plantlet in vitro, by day lake gold Line lotus plantlet in vitro is implanted on the kind planting-line in forest land, and day lake roxburgh anoectochilus terminal bud plantlet in vitro plantation spacing is 3-5cm, planting depth preferably it is shallow not It is preferably deep, it is advisable with hiding a day root for lake roxburgh anoectochilus terminal bud;With carbendazim or thiram 1000 after its lake roxburgh anoectochilus terminal bud plantlet in vitro transplanting Times water dilute solution is poured as root water;Implantation time is advisable with the annual 4-8 months, as long as other times temperature is higher than 10 DEG C Also can plant, the present embodiment is in June.
5. forest management:Day lake roxburgh anoectochilus terminal bud ecosystem kind woodlot relative air humidity is kept to be maintained at more than 70%, and Shi Qingli fallen leaves ensure that day lake anoectochilus blume seedling can receive the scattered light of abundance.In day lake roxburgh anoectochilus terminal bud plantlet in vitro growth period, when When kind planting-line is more dried, water spray process is carried out using water injection system so that plant planting-line and air keeps the state of moistening.Plant Day lake roxburgh anoectochilus terminal bud after one month, with solution as Foliar Spraying, sprays to the dropping liquid of blade face, applies per 7~10 days later Fertilizer (with solution as Foliar Spraying) once, in disease season occurred frequently, is made every effort to overcome with agricultural streptomycin sulphate 100mg/kg or general The rotations such as 1000 times of water diluents, 1000 times of water diluents of carbendazim are sprayed, and are monthly sprayed 1 time, and snail is multiple with slug to be born in 5~ In June, spread fertilizer over the fields in surrounding preventing and treating with quick lime;Described solution be per 15 jin of peanut press pulps plus 150 jin of water spontaneous fermentation 30 days after 8 times of liquid fertilizer dilute with water after, then per liter adds 2.0 grams of spend precious No. 1 foliage fertilizer, 1.7 grams of potassium dihydrogen phosphates to be well mixed Form solution.
6. harvest:In day lake, roxburgh anoectochilus terminal bud plantlet in vitro bottle outlet is colonized 7~9 months and can harvest, now day lake roxburgh anoectochilus terminal bud plant Harvested by growing to 6~more than 10cm of height, 5~6 leaves, every plant of 2~3g of fresh weight.Day lake roxburgh anoectochilus terminal bud wildness now Shape is good, and using effect is good, drug effect height (identified, it is identical with wild day lake roxburgh anoectochilus terminal bud), thus its price is planted than Artificial facilities The day lake roxburgh anoectochilus terminal bud of training wants high by more than 1/3rd.
Embodiment 2:
1. Proterozoic artificial pollination:In the roxburgh anoectochilus terminal bud Proterozoic of Conghua of Guangdong Province day lake (37 points to 23 degree 40 points of north latitude 23 degree, 36 points to 113 degree 38 points of east longitude 113 degree), choose robust growth, stem is thick, blade is big, blade reticulate pattern is clear and many, resistance, resist The strong day lake roxburgh anoectochilus terminal bud of pest and disease damage (Anoectochilus roxburghii (wall) lindl. ' tianhu ') health strain, in Its lake roxburgh anoectochilus terminal bud flowering naturally season carries out the information such as artificial pollination and bagging, mark date, and fruit grows under natural conditions Development.
2. aseptic seeding produces seedling:It is explant to choose the roxburgh anoectochilus terminal bud capsule that fruit age is 50~70 days, in volume fraction Soak 30 seconds in 70% alcohol water blend, then with the water-soluble liquid disinfectant of the mercuric chloride of mass fraction 0.1% 15~20 minutes, aseptic water washing 4~5 times, the moisture on capsule surface is blotted with aseptic filter paper under aseptic condition on superclean bench.Then aseptic operation knife is used Capsule is cut, [described embryo germination and protocorm divides for uniform broadcasting to embryo germination and protocorm differentiation growth medium A1 Long per liter of the culture medium A 1 of metaplasia is to be configured so that:Treasured No. 1 2g, peptone 0.5g, activated carbon 100mg, inositol 80mg, sweet ammonia will be spent Sour 2.5mg, thiamine hydrochloride (VB1) 0.05mg, puridoxine hydrochloride (VB6) 0.8mg, nicotinic acid 0.4mg, methyl α-naphthyl acetate (NAA) 0.5mg, sucrose 15g, agar 7g, in being added to a small amount of water, are then settled to 1L with water, adjust pH 5.6].After 90 days, plant Embryo expands and to form continued growth after protocorm and form high 2~3 centimetres of seedlings with root.Embryo germination and protocorm differentiation growth step Section cultivation temperature be (23 ± 2) DEG C, 500~1000lux of illuminance, illumination 12h/d.By high 2~3 centimetres of seedling, go to [per liter of Rooting and hardening-off culture base A2 is configured so that Rooting and hardening-off culture base A2:Spend No. 1 2g of treasured, peptone 0.5g, mashed potatoes 80g, activated carbon 500mg, inositol 120mg, glycine 1.5mg, thiamine hydrochloride (VB1) 0.2mg, puridoxine hydrochloride (VB6) 0.4mg, nicotinic acid 0.8mg, methyl α-naphthyl acetate (NAA) 0.2mg, sucrose 30g, agar 6g, in being added to a small amount of water, are then settled to water 1L, adjusts pH 5.4], during culture 90 days, height of seedling can reach 5~7 centimetres, radical 3~5, and rooting rate is 100%.Strong plantlets and rootage The cultivation temperature in stage be (25 ± 2) DEG C, 1000~2000lux of illuminance, illumination 12h/d.Roxburgh anoectochilus terminal bud seedling of taking root in day lake is existed Natural light lower refining seedling bottle outlet after 7~15 days.Before plantation, seedling of taking root is taken out from blake bottle, after cleaning the culture medium of attachment, use thousand The liquor potassic permanganate of/mono- (mass fraction) soaks 5~10min, and a day lake roxburgh anoectochilus terminal bud plantlet in vitro is obtained after draining, and waits to plant. To go up to the sky, lake seedlings of anoectochilus are produced near to the tissue culture workshop that day lake side is set up.The genuine roxburgh anoectochilus terminal bud capsule in one day lake leads to Cross the aseptic seeding culture of 6~August, can quick propagating and cultivating go out more than 100,000 plants of day lake roxburgh anoectochilus terminal bud plantlet in vitro.
3. forest land selects and site preparation:The genuine roxburgh anoectochilus terminal bud Proterozoic in day lake is selected, height above sea level is in the subtropical zone of 100-500m Mountain region or hills forest land, 20.9 degree of year-round average temperature, 38.7 degree of maximum temperature, minimum temperature -2.5 is spent;Average annual rainfall 2100 milliliters, precipitation concentrates the 4-8 months;Vegetation is green throughout the year based on evergreen broadleaf forest and theropencedrymion, well arranged, strongly fragrant In 0.85-0.9, periphery has sufficient pollution-free water source to degree of closing, and long-term air humidity is more than 60%;In nearly small stream limes marginis shady place woods Ground, removes forest land weeds and surface fallen leaves, and the soil layer of sylvan life is turned over, and ploughed depth is 20-30cm, with the soil layer that ravels, And the kind planting-line of 5 meters of wide 60 centimeter length is played into, the fertile fertile soil that layer overlay periphery hillside fields is collected on furrow;Two meter ampere on furrow Dress fallen leaves bearer network, carries the blade that trees drop, it is to avoid cover the day lake roxburgh anoectochilus terminal bud in forest land;Water spray system is installed simultaneously on furrow System, can carry out spray irrigation under more dry weather, keep the humidity of air.
4. the plantation of day lake roxburgh anoectochilus terminal bud plantlet in vitro:Using sterile-processed day lake roxburgh anoectochilus terminal bud plantlet in vitro, by day lake gold Line lotus plantlet in vitro is implanted on the kind planting-line in forest land, and day lake roxburgh anoectochilus terminal bud plantlet in vitro plantation spacing is 3-5cm, planting depth preferably it is shallow not It is preferably deep, it is advisable with hiding a day root for lake roxburgh anoectochilus terminal bud;With carbendazim or thiram 1000 after its lake roxburgh anoectochilus terminal bud plantlet in vitro transplanting Times water dilute solution is poured as root water;Implantation time is advisable with the annual 4-8 months, as long as other times temperature is higher than 10 degree Also can plant, the present embodiment is in June.
5. forest management:Day lake roxburgh anoectochilus terminal bud ecosystem kind woodlot air humidity is kept to be maintained at more than 70%, it is clear in time Reason fallen leaves ensure that day lake anoectochilus blume seedling can receive the scattered light of abundance.In day lake roxburgh anoectochilus terminal bud plantlet in vitro growth period, work as plantation When furrow are more dried, water spray process is carried out using water injection system so that plant planting-line and air keeps the state of moistening.Plant one Day lake roxburgh anoectochilus terminal bud after month, with solution as Foliar Spraying, sprays to the dropping liquid of blade face, and later per 7~10 days, fertilising (was used Solution is used as Foliar Spraying) once, in disease season occurred frequently, with agricultural streptomycin sulphate 100mg/kg or general 1000 times are made every effort to overcome The rotations such as water diluent, 1000 times of water diluents of carbendazim are sprayed, and are monthly sprayed 1 time, and snail is born in for 5~June with slug is multiple, Spread fertilizer over the fields in surrounding preventing and treating with quick lime;Described solution be per 15 jin of peanut press pulps plus 150 jin of water spontaneous fermentation 30 days after liquid fertilizer After 8 times of dilute with water, then per liter adds 2.0 grams of spend precious No. 1 foliage fertilizer, 1.7 grams of potassium dihydrogen phosphates to be well mixed to form fertilizer Liquid.
6. harvest:In day lake, roxburgh anoectochilus terminal bud plantlet in vitro bottle outlet is colonized 7~9 months and can harvest, now day lake roxburgh anoectochilus terminal bud plant Harvested by growing to 6~more than 10cm of height, 5~6 leaves, every plant of 2~3g of fresh weight.Day lake roxburgh anoectochilus terminal bud wildness now Shape is good, and using effect is good, drug effect height (identified, it is identical with wild day lake roxburgh anoectochilus terminal bud), thus its price is planted than Artificial facilities The day lake roxburgh anoectochilus terminal bud of training wants high by more than 1/3rd.

Claims (4)

1. genuine roxburgh anoectochilus terminal bud ecosystem implantation methods in a kind of Conghua of Guangdong Province day lake, it is characterised in that comprise the following steps:
The acquisition of A, day lake roxburgh anoectochilus terminal bud plantlet in vitro:In the roxburgh anoectochilus terminal bud Proterozoic of Conghua of Guangdong Province day lake, choose robust growth, stem it is thick, The day lake roxburgh anoectochilus terminal bud health strain that blade is big, blade reticulate pattern is clear and many, resistance, disease and insect resistance are strong, in day lake roxburgh anoectochilus terminal bud certainly So season of flowers carries out artificial pollination and bagging, and fruit grows under natural conditions, chooses the capsule that fruit age is 50~70 days It is really explant, the sterilized moisture for blotting capsule surface with aseptic filter paper under aseptic condition on superclean bench, then Capsule is cut with aseptic operation knife, embryo germination and protocorm differentiation growth medium A1 is broadcasted sowing, in 23 ± 2 DEG C, illuminance 500~1000lux, cultivates under illumination 12h/d, and embryo expands to form seedling of the continued growth formation with root after protocorm, will be high by 2 ~3 centimetres of seedling, goes to Rooting and hardening-off culture base A2 cultures, and cultivation temperature is 25 ± 2 DEG C, 1000~2000lux of illuminance, Illumination 12h/d, obtains day lake roxburgh anoectochilus terminal bud and takes root seedling, and day lake roxburgh anoectochilus terminal bud is taken root seedling bottle outlet after natural light lower refining seedling 7~15 days, Bottle seedling of taking root is obtained, before plantation, is taken out from blake bottle and is taken root bottle seedling, after cleaning the culture medium of attachment, with thousand points of mass fraction One of potassium permanganate solution soak 5~10min, a day lake roxburgh anoectochilus terminal bud plantlet in vitro is obtained after draining, wait to plant;Described embryo Sprout and protocorm differentiation growth medium A1, its per liter containing spending No. 1 1~2g of treasured, 0.5~2g of peptone, activated carbon 50~ 100mg, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2mg of thiamine hydrochloride, puridoxine hydrochloride 0.4~ 0.8mg, 0.4~0.8mg of nicotinic acid, 0.2~0.5mg of methyl α-naphthyl acetate, 15~20g of sucrose, 6~7g of agar, balance of water, pH 5.4- 5.6;Described Rooting and hardening-off culture base A2, its per liter containing spending treasured No. 1 1~2g, 0.5~2g of peptone, 40~80g of mashed potatoes, 500~1000mg of activated carbon, 80~120mg of inositol, 1.5~2.5mg of glycine, 0.05~0.2mg of thiamine hydrochloride, hydrochloric acid pyrrole Tremble 0.4~0.8mg of alcohol, 0.4~0.8mg of nicotinic acid, 0.2~0.5mg of methyl α-naphthyl acetate, 20~30g of sucrose, and 6~7g of agar is balance of Water, pH 5.4-5.6;
B, forest land select and site preparation:Select day lake roxburgh anoectochilus terminal bud Proterozoic, Subtropical Mountainous or mound of the height above sea level in 100-500m Mound forest land, vegetation is green throughout the year based on evergreen broadleaf forest and theropencedrymion, well arranged, and canopy density are in 0.85-0.9, week While there is sufficient pollution-free water source, long-term air humidity is more than 60%;In nearly small stream limes marginis shady place forest land, remove forest land weeds and Surface is fallen leaves, and the soil layer of sylvan life is turned over, and ploughed depth is 20-30cm, with the soil layer that ravels, and plays kind of a planting-line, furrow upper berth The fertile fertile soil that last layer periphery hillside fields is collected;Fallen leaves bearer network is installed on furrow, water injection system is installed simultaneously on furrow;
The plantation of C, day lake roxburgh anoectochilus terminal bud plantlet in vitro:When the annual 4-8 months or other month temperatures being higher than 10 DEG C, by day lake gold Line lotus plantlet in vitro is implanted on the kind planting-line in forest land, and day lake roxburgh anoectochilus terminal bud plantlet in vitro plantation spacing is 3-5cm, and planting depth is soil layer A day root for lake roxburgh anoectochilus terminal bud is hidden, root water is poured;
D, forest management:Using water injection system water spray keep day lake roxburgh anoectochilus terminal bud kind woodlot relative air humidity be maintained at 70% with On, cleaning fallen leaves, in day lake roxburgh anoectochilus terminal bud plantlet in vitro growth period, water spray process is carried out using water injection system so that plant planting-line and Air keeps the state of moistening, applies fertilizer after plant, and prevention and elimination of disease and pests;
E, harvesting:Day lake roxburgh anoectochilus terminal bud is harvested to collecting season.
2. genuine roxburgh anoectochilus terminal bud ecosystem implantation methods in Conghua of Guangdong Province day according to claim 1 lake, it is characterised in that described The step of A sterilization be to soak explant 30 seconds in the alcohol of volume fraction 70%, then with 0.1% mercuric chloride solution sterilization 15~ 20 minutes, aseptic water washing 4~5 times.
3. genuine roxburgh anoectochilus terminal bud ecosystem implantation methods in Conghua of Guangdong Province day according to claim 1 lake, it is characterised in that described The step of C pouring root water be to be made with 1000 times of water dilute solutions of carbendazim or thiram after the roxburgh anoectochilus terminal bud plantlet in vitro transplanting of day lake For root water pouring.
4. genuine roxburgh anoectochilus terminal bud ecosystem implantation methods in Conghua of Guangdong Province day according to claim 1 lake, it is characterised in that described The step of D plant after apply fertilizer, and prevention and elimination of disease and pests be transplant one month from day lake roxburgh anoectochilus terminal bud plantlet in vitro after, with solution as leaf Leaven is sprayed, and is sprayed to the dropping liquid of blade face, and later per 7~10 days, fertilising once, in disease season occurred frequently, used agricultural sulfate chain Mycin 100mg/kg or general makes every effort to overcome the rotations such as 1000 times of water diluents, 1000 times of water diluents of carbendazim and sprays, and monthly sprays 1 time, Snail is born in for 5~June with slug is multiple, is spread fertilizer over the fields in surrounding preventing and treating with quick lime;Described solution is added per 15 jin of peanut press pulps After 8 times of liquid fertilizer dilute with water after 150 jin of water spontaneous fermentation 30 days, then per liter of precious No. 1 foliage fertilizer of spending for adding 2.0 grams, 1.7 Gram potassium dihydrogen phosphate, is well mixed to form solution.
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