Background technology
Chloranthus glaber (Sarcandra glabra (Thunb.) Nakai), another name: Glabrous Sarcandra Herb, synthetism lotus, sarcandra, diffuse dayflower herb etc. are Chloranthaceae Sarcandra perennial evergreen fruticuli.The complete stool hyoscine, have anti-inflammation, clearing heat and detoxicating, dispel rheumatism, the effect such as promoting blood circulation and stopping pain, the synthetism of stimulating the menstrual flow, be used for the treatment of various diseases associated with inflammation, rheumatic arthritis, sore swollen toxin, traumatic injury, fracture etc., remarkable to the Several Kinds of Malignancy effect, be the good traditional Chinese medicine that a kind of pharmacological action is extensive, toxicity is low.Along with continually developing of the new product take Chloranthus glaber as raw material, increasing to its demand, traditional complete stool mode of excavating causes going to wreck property of wild resource destruction, causes raw medicinal material in short supply, so Chloranthus glaber has boundless plantation DEVELOPMENT PROSPECT.Under natural conditions, Chloranthus glaber is born in hillside, cheuch sylvan life dark and damp place, 420~1500 meters of height above sea level.The environment of happiness warm moist is avoided high light direct projection and long high-temperature drying.The habit of growth of Chloranthus glaber uniqueness requires planting process that certain condition of sheltering from heat or light must be provided, and the measure of manually sheltering from heat or light will improve planting cost greatly.If Chloranthus glaber is interplanted in mao bamboo woods, can effectively address this problem.
Mao bamboon is distributed in 23 ° 23 ' ~ 32 ° 20 ' of China's north latitude, in the broad regions that east longitude is 104 ° 30 ' ~ 122 °, is that China's distribution is the widest, area is maximum, one of economic, ecological, that social benefit is best bamboo kind.There are the mao bamboon cultivation in/3rd above provinces and regions, the whole nation, municipality directly under the Central Government at present, and existing mao bamboo woods area is 2,800,000 hm nearly
2, wherein major part is the material mao bamboo woods that Management Technology is simple and input-output ratio is high.And the continuous expansion of the downslide of bamboo shoots price in recent years and bamboo wood range of application makes material further increase with the cultivated area of woods.But single plantation mao bamboon economic benefit is limited, and interplant Chloranthus glaber in mao bamboo woods, not only save the link of shading in the Chloranthus glaber cultivation, reduced planting cost, the special odor and the efficacy of a drug that also have because of Chloranthus glaber can reduce damage by disease and insect, the more important thing is conditions such as to take full advantage of soil and light, temperature, water, fertilizer, the output of raising the multiple-cropping index and obviously increase economic benefit.
In addition, interplant on a large scale Chloranthus glaber and must first solve the seedling breeding problem, its traditional modes of reproduction has cuttage, sowing and plant division etc., but reproduction coefficient is low, survival rate is low, seedling cost is relatively high and be subject to weather influence.Utilizing the Plant Tissue Breeding means to set up the quick reproduction technique system is optimal selection.Publication number be CN 101194593A Introduction To Cn Patent a kind of tissue culture and rapid propagation method of Chloranthus glaber, the method obtains regrowth take the root system of aseptic seedling as explant passes through the callus approach, but the method must induce the test-tube plantlet of acquisition as the examination material take Chloranthus glaber aseptic seeding or stem segment with axillary buds, complex steps, production cycle are long, and still face Chloranthus glaber seed maturity rate and utilization percent is low, seed source is difficult, drawing materials is subject to seasonal restrictions the problem low with germination rate.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, the method for ecological interplanting Chloranthus glaber under a kind of mao bamboo woods is provided, with planting cost, the cultivation period that shortens Chloranthus glaber that reduces Chloranthus glaber, the output that improves Chloranthus glaber.
The scheme that the present invention solve the technical problem is: a kind of method of ecological interplanting Chloranthus glaber under mao bamboo woods is characterized in that: comprise the steps,
(1) tissue culture
Choose the Chloranthus glaber blade and make explant, after sterilization, blade is cut into small pieces and is placed on the adventitious bud induction culture base, inoculate to transfer to after 5 days and continue in fresh adventitious bud induction culture base to cultivate 30~35 days, the adventitious bud induction culture base is take MS as minimal medium, additional 2.0~4.0 mg/L TDZ, 0.1~0.2 mg/L NAA, 100~150g/L banana puree and 0.5~1.0g/L active carbon;
The indefinite bud that blade edge is grown is cut into single bud, is inoculated in proliferated culture medium, and proliferated culture medium is take MS as minimal medium, additional 1.0~2.0 mg/L TDZ and 0.05~0.1 mg/L NAA;
Cut the simple bud of high 2~3cm, be inoculated in root media, root media is take 1/2MS as minimal medium, additional 0.1~0.2 mg/L NAA;
Above all medium all add 6.0 mg/L agar and 30.0 mg/L sucrose, and it is 25 ± 2 ℃ of temperature, illuminance 1500~2000 lx that group is trained the chamber condition of culture, illumination every day 12 h;
(2) hardening
Bottle seedling in late April culture of rootage about 20 days is transported to the mao bamboo woods lower refining seedling, does not open the bottle cap hardening 10~12 days, then opens the bottle cap hardening 2~3 days;
(3) transplant
After hardening is completed, group training transplantation of seedlings is the north slope bamboo grove of 150~400 strain/mus or the southern slope bamboo grove that vertical bamboo degree is 200~400 strain/mus in the vertical bamboo degree of mao bamboon;
(4) forest management
Use in conjunction with bamboo grove and urge whip fertilizer to intertill for the first time annual June, dark 10~15cm, and apply urea or the composite fertilizer of 5kg/ mu, intertill September for the second time; Field planting then winter the ground mulching straw around the root;
(5) gather
The field planting Second Year plays the Chloranthus glaber of gathering annual 11~December, and completes before cutting down bamboo and gather.
As further technical scheme, in described transplant step, the Chloranthus glaber seedling is vertically in a row planted along the hillside physical features, spacing in the rows 35~40cm, two row's spacing 25~30cm arrange a pavement that 1.0~1.2m is wide every 6~8 Lysimachia sikokiana corals.
As technical scheme further, after described group of training transplantation of seedlings at its foliage-spray 0.1% calcium chloride.
As technical scheme further, the type of described bamboo grove is the material mao bamboo woods.
Beneficial effect of the present invention has:
(1) at mao bamboon forest interplanting Chloranthus glaber, need not occupy cultivated land separately, not need prior afforestation, improve land utilization rate, reduce the planting cost of Chloranthus glaber; Mao bamboo woods are the Chloranthus glaber moisturizing of sheltering from heat or light, thereby make Chloranthus glaber not be subjected to the good growth of direct sunlight, and the special odor that Chloranthus glaber has and the efficacy of a drug can reduce the damage by disease and insect of mao bamboo woods again, make two kinds of crop harmonic growths, form benign cycle; On the other hand, in the time of the farming Chloranthus glaber, bamboo grove is also fostered, be conducive to the management of mao bamboo woods, changed the traditional single cropping pattern of mao bamboo woods, can greatly improve farmers' income;
(2) the present invention makes explant with the Chloranthus glaber blade, set up a kind of Chloranthus glaber blade high-efficiency regeneration system, by the method carry out that seedling breeding is simple and easy to do, efficient stable, material source is abundant and endophytic bacterial contamination is few, can obtain at short notice the seedling that a large amount of germplasms are good, proterties is unified, for extensive, standardization interplanting Chloranthus glaber provide powerful guarantee;
(3) with hardening off method hardening of the present invention, and in conjunction with suitable transplanting method, Chloranthus glaber group training seedling can directly be transplanted to mao bamboo woods, does not need to have saved the time through the nursery phase, has also saved the cost that construction of nursery garden, management and secondary are transplanted.
(4) the present invention is along the vertical in a row plantation Chloranthus glaber of hillside physical features, and every 6~8 rows, a pavement is set, and vacant lot is provided not only for forest management, operation, also convenient felling mao bamboon operation and mao bamboon is pulled.
(5) bamboo shoot are not suitable for interplanting Chloranthus glaber with woods or the dual-purpose woods of bamboo shoot material, because it need to dig in a large number bamboo shoot and cultivate again easy damaged sylvan life crop during farming every year.The type of bamboo grove of the present invention is the material mao bamboo woods, can reduce to greatest extent personnel's activity to the impact of Chloranthus glaber.
Embodiment
The present invention will be further described below in conjunction with embodiment, and test is carried out under the mao bamboo woods of Zhejiang Mountainous Region, has selected 4 different material mao bamboo woods, tests simultaneously.
Embodiment 1:
(1) tissue culture
Choose the healthy disease-free tender leaf of Chloranthus glaber and make explant, first use 75% alcohol-pickled 20s in superclean bench, then soak 1min with 0.1% mercuric chloride solution, aseptic water washing 3~4 times is cut into approximately 1cm of area
2The blockage of left and right, standby.Blade with after sterilization lies against in the adventitious bud induction culture base up with adaxial and its surface, inoculates to transfer to after 5 days and continues in fresh culture to cultivate 30 days.The adventitious bud induction culture base is take MS as minimal medium, additional 2.5 mg/L TDZ, 0.15 mg/L NAA, 120g/L banana puree and 0.8g/L active carbon; After 15 days, incision slightly has expands and has green bud point to occur blade inoculation, begins subsequently to have the regeneration of indefinite bud to fresh culture.When cultivating 30 days, explant regeneration frequency and average regenerated adventitious bud number are respectively 82.1% and 5.3.The indefinite bud that blade edge is grown is cut into single bud, is inoculated in proliferated culture medium, and proliferated culture medium is take MS as minimal medium, additional 1.2mg/L TDZ and 0.06 mg/L NAA.Available identical proliferated culture medium is shoot proliferation repeatedly, every 25~30 days subcultures once, the average propagation multiple of Multiple Buds is 6.2.Cut the simple bud of high 2~3cm, be inoculated in root media, root media is take 1/2MS as minimal medium, additional 0.15 mg/L NAA.Begin to take root about 12 days, when being cultured to 20 days, rooting rate 100%.The time of beginning culture of rootage decides according to transplanting time.Above all medium all add 6.0 mg/L agar and 30.0 mg/L sucrose, and it is 25 ± 2 ℃ of temperature, illuminance 1500~2000 lx that group is trained the chamber condition of culture, illumination every day 12 h.
(2) hardening
In late April, 2009, the bottle seedling with culture of rootage about 20 days is transported under traffic convenience, the water source is convenient, physical features is smooth mao bamboo woods and concentrates hardening.The bottle seedling is neatly put on ground by bottle spacing 5cm * 5cm, first do not opened the bottle cap hardening 10 days, then opened the bottle cap hardening 3 days, make group training seedling progressively adapt to the plantation environment.After hardening is completed, group training seedling presss from both sides out with tweezers, and water washes away after the root medium minute to be delivered to peripheral forest land and transplant.
(3) transplant
Begin whole ground the first tenday period of a month in May to the middle ten days and transplant in the material mao bamboo woods that are positioned at north slope, the vertical bamboo degree of north slope bamboo grove mao bamboon is controlled at 150~400 strain/mus, always cultivated area is 2.5 mu.Surgery weeds, shrub in woods, dig up the bamboo root and stem of certain plants, shrub is first-class, then plough, flatten.The Chloranthus glaber seedling is in a row planted along the hillside physical features is vertical, spacing in the rows 35~40cm, two row's spacing 25~30cm after every plantation 6~8 Lysimachia sikokiana corals, reserve the wide pavement of 1.0~1.2m, so that cut down the mao bamboon operation and mao bamboon is pulled; Can distribute according to bamboo during plantation slightly adjusts, and needn't too impose rule standardized.Dig cave, the dark 10~15cm in cave during transplanting, the table soil of humus is rich in backfill on a small quantity, but the cave deeply still should be less than 5cm, and plant is put into the cave, covers root system, compacting with table soil as far as possible.Irrigate normal root water, foliage-spray transpiration inhibitor 0.1% calcium chloride, spray once, connects and sprays 3 times week about.Need in time water as the weather arid.Transplant and can water for the first time the night soil that becomes thoroughly decomposed in rear about one month, the dilution ratio of night soil and water is 1: 15, uses later on night soil in good time.In the time of one month, the statistics transplanting survival rate is 93%.
(4) forest management
Use in conjunction with bamboo grove and urge whip fertilizer to intertill for the first time annual June, dark 10~15cm, and apply the urea of 5kg/ mu, and intertill September for the second time, weeds a garden, and keeps loosing soil, and operating process takes into account mao bamboon and fosters; Keep ground moistening vegetative period, in field planting summer then, as surpassing the not rainfall of 2 weeks, need in time water.In December, 2009, the ground mulching straw around the root, but both heat-insulating and colds can promote that again the beginning of spring, plant early gave birth to fast long.Be buried in the earth straw as fertilizer sources from root 20cm place trench digging in March, 2010.
(5) gather
The Chloranthus glaber of gathering in December, 2010, in December, 2011 and in November, 2012 selects fine day to gather, and extracts cauline leaf on the ground, cut stay greatly little, allow branch tenderer, length≤20cm stay continued growth, and long, older, ripe stem branch, cut off at distance basal part of stem 5~10cm place.Play Chloranthus glaber growth November and stop gradually, blade is dark green, fruit reddens, this moment biomass reach the highest, the harvesting optimum; Cut down again mao bamboon and harvest Chloranthus glaber, not only meet the management rule of bamboo grove but also make the felling process convenient.Can gather in once every year later on.The disposable plantation of Chloranthus glaber, income for many years, the circulation results reach 20 years.
The medicinal material of harvesting in 3 years all is sent to city medicine inspecting institute and detects, and Isofraxidin content is 0.031~0.035% after testing, reaches standards of pharmacopoeia (must not be less than 0.020%) fully.Obtained respectively Chloranthus glaber dry product 525,852 and 956 kilograms in 3 years, amount to into yield per unit area and be respectively 210,340.8 and 382.4 thousand g/acres, cumulative year after year.
From between 2009 to 2012 4 years, any damage by disease and insect does not occur in mao bamboon, and the average diameter of a cross-section of a tree trunk 1.3 meters above the ground of newborn mao bamboon has increased 10% left and right.
Embodiment 2:
(1) tissue culture
Choose the healthy disease-free tender leaf of Chloranthus glaber and make explant, first use 75% alcohol-pickled 20s in superclean bench, then soak 1min with 0.1% mercuric chloride solution, aseptic water washing 3~4 times is cut into approximately 1cm of area
2The blockage of left and right, standby.Blade with after sterilization lies against in the adventitious bud induction culture base up with adaxial and its surface, inoculates to transfer to after 5 days and continues in fresh culture to cultivate 35 days.The adventitious bud induction culture base is take MS as minimal medium, additional 2.0 mg/L TDZ, 0.1 mg/L NAA, 150g/L banana puree and 0.5g/L active carbon; After 17 days, incision slightly has expands and has green bud point to occur blade inoculation, begins subsequently to have the regeneration of indefinite bud to fresh culture.When cultivating 30 days, explant regeneration frequency and average regenerated adventitious bud number are respectively 80.2% and 4.9.The indefinite bud that blade edge is grown is cut into single bud, is inoculated in proliferated culture medium, and proliferated culture medium is take MS as minimal medium, additional 1.0mg/L TDZ and 0.1 mg/L NAA.Available identical proliferated culture medium is shoot proliferation repeatedly, every 25~30 days subcultures once, the average propagation multiple of Multiple Buds is 5.7.Cut the simple bud of high 2~3cm, be inoculated in root media, root media is take 1/2MS as minimal medium, additional 0.2 mg/L NAA.Begin to take root about 14 days, when being cultured to 20 days, rooting rate 100%.The time of beginning culture of rootage decides according to transplanting time.Above all medium all add 6.0 mg/L agar and 30.0 mg/L sucrose, and it is 25 ± 2 ℃ of temperature, illuminance 1500~2000 lx that group is trained the chamber condition of culture, illumination every day 12 h.
(2) hardening
In late April, 2009, the bottle seedling with culture of rootage about 20 days is transported under traffic convenience, the water source is convenient, physical features is smooth mao bamboo woods and concentrates hardening.The bottle seedling is neatly put on ground by bottle spacing 5cm * 5cm, first do not opened the bottle cap hardening 12 days, then opened the bottle cap hardening 2 days, make group training seedling progressively adapt to the plantation environment.After hardening is completed, group training seedling presss from both sides out with tweezers, and water washes away after the root medium minute to be delivered to peripheral forest land and transplant.
(3) transplant
Begin whole ground the first tenday period of a month in May to the middle ten days and transplant in the material mao bamboo woods that are positioned at Nan Po, the vertical bamboo degree of southern slope bamboo grove mao bamboon is controlled at 200~400 strain/mus, always cultivated area is also 2.5 mu.Surgery weeds, shrub in woods, dig up the bamboo root and stem of certain plants, shrub is first-class, then plough, flatten.The Chloranthus glaber seedling is in a row planted along the hillside physical features is vertical, spacing in the rows 35~40cm, two row's spacing 25~30cm after every plantation 6~8 Lysimachia sikokiana corals, reserve the wide pavement of 1.0~1.2m, so that cut down the mao bamboon operation and mao bamboon is pulled; Can distribute according to bamboo during plantation slightly adjusts, and needn't too impose rule standardized.Dig cave, the dark 10~15cm in cave during transplanting, the table soil of humus is rich in backfill on a small quantity, but the cave deeply still should be less than 5cm, and plant is put into the cave, covers root system, compacting with table soil as far as possible.Irrigate normal root water, foliage-spray transpiration inhibitor 0.1% calcium chloride, spray once, connects and sprays 3 times week about.Need in time water as the weather arid.Transplant and can water for the first time the night soil that becomes thoroughly decomposed in rear about one month, the dilution ratio of night soil and water is 1: 20, uses later on night soil in good time.In the time of one month, the statistics transplanting survival rate is 92%.
(4) forest management
Use in conjunction with bamboo grove and urge whip fertilizer to intertill for the first time annual June, dark 10~15cm, and apply the composite fertilizer of 5kg/ mu, and intertill September for the second time, weeds a garden, and keeps loosing soil, and operating process takes into account mao bamboon and fosters; Keep ground moistening vegetative period, in field planting summer then, as surpassing the not rainfall of 2 weeks, need in time water.In December, 2009, the ground mulching straw around the root, but both heat-insulating and colds can promote that again the beginning of spring, plant early gave birth to fast long.Be buried in the earth straw as fertilizer sources from root 20cm place trench digging in March, 2010.
(5) gather
The Chloranthus glaber of gathering in December, 2010, in December, 2011 and in November, 2012 is gathered to be operated in and is completed before cutting down bamboo, and results situation and embodiment 1 are without significant difference.
Embodiment 3:
The operating procedure of the present embodiment and parameter are with reference to embodiment 1, and different is: blade adventitious bud induction culture base is take MS as minimal medium, additional 4.0 mg/L TDZ, 0.2 mg/L NAA, 100g/L banana puree and 1.0g/L active carbon; After 15 days, incision slightly has expands and has green bud point to occur blade inoculation, begins subsequently to have the regeneration of indefinite bud to fresh culture.When cultivating 30 days, explant regeneration frequency and average regenerated adventitious bud number are respectively 78.4% and 4.7.The indefinite bud that blade edge is grown is cut into single bud, is inoculated in proliferated culture medium, and proliferated culture medium is take MS as minimal medium, additional 2.0mg/L TDZ and 0.05 mg/L NAA.Available identical proliferated culture medium is shoot proliferation repeatedly, every 25~30 days subcultures once, the average propagation multiple of Multiple Buds is 6.0.Cut the simple bud of high 2~3cm, be inoculated in root media, root media is take 1/2MS as minimal medium, additional 0.1mg/L NAA.Begin to take root about 14 days, when being cultured to 20 days, rooting rate 100%.After group training seedling upgrowth situation, transplanting, survival rate and results situation and embodiment 1 are without significant difference.
Embodiment 4:
The operating procedure of the present embodiment and parameter are with reference to embodiment 2, and different is: select the following southern slope bamboo grove of vertical bamboo degree 200 strain/mus during transplanting.Plant after 1 month, transplanting survival rate is only 55% left and right.In process of growth, because solar radiation has the dried-up phenomenon in blade face, the severe patient complete stool is dead.In the autumn of plantation Second Year, the Chloranthus glaber survival rate is 20% left and right, and plant is short and small, leaf color is lighter, partially yellow, gather in.