CN105875412A - Planting method of organic selenium-rich sweet Gynostemma pentaphylla - Google Patents
Planting method of organic selenium-rich sweet Gynostemma pentaphylla Download PDFInfo
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- CN105875412A CN105875412A CN201610271020.1A CN201610271020A CN105875412A CN 105875412 A CN105875412 A CN 105875412A CN 201610271020 A CN201610271020 A CN 201610271020A CN 105875412 A CN105875412 A CN 105875412A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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Abstract
The invention provides a planting method of organic selenium-rich sweet Gynostemma pentaphylla, and belongs to the technical field of cultivation. The planting method of the organic selenium-rich sweet Gynostemma pentaphylla comprises the following steps: selecting varieties of the sweet Gynostemma pentaphylla, carrying out tissue culture on explants of the sweet Gynostemma pentaphylla at first to obtain tissue culture seedlings; then transplanting the tissue culture seedlings in a nursery garden to build a strain garden; planting rhizomes of plants of Gynostemma pentaphylla growing for 1 year in the strain garden to build a propagation expanding garden; planting rhizomes of plants of the Gynostemma pentaphylla growing for 1 year in the propagation expanding garden to build a garden for a land for growing field crops; and planting rhizomes of plants of the Gynostemma pentaphylla growing for 1 year in the garden for the land for growing field crops to the land for growing field crops. The strain garden is built by tissue culture technology, the sweet Gynostemma pentaphylla can be quickly reproduced by tissue culture, the yield of the sweet Gynostemma pentaphylla is increased while viral diseases are reduced, then by building of three-garden reproduction, the tissue culture seedlings subjected to tissue culturing grow well, the quality and yield of the sweet Gynostemma pentaphylla are increased, and high-yield and high-efficiency planting is realized.
Description
Technical field
The present invention relates in technical field of cultivation, in particular to a kind of organic selenium-rich sugariness
The implantation methods of Herb Gynostemmae Pentaphylli.
Background technology
Whole world Herb Gynostemmae Pentaphylli has 16 kinds and 3 mutation, and there are 15 kinds and 3 mutation in China, only
Just there are 11 kinds and 3 mutation in state, enshi.The former mutation of Herb Gynostemmae Pentaphylli promoted both at home and abroad is because of taste
Extremely bitter, cold heavy, it is difficult to entrance, injuring spleen-stomach, become Herb Gynostemmae Pentaphylli artificial growth and industrialization
The bottleneck of exploitation；In addition, Herb Gynostemmae Pentaphylli is in wild and semi-wild state for a long time, and domestic do not have
There is the kind being available for cultivation, be suitable for its growth and realize the high-yield high-efficiency cultivation skill of its health value
Art research is blank.
Summary of the invention
It is an object of the invention to provide the implantation methods of a kind of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, no
Only keep the kind of sugariness Herb Gynostemmae Pentaphylli, improve the quality and yield of sugariness Herb Gynostemmae Pentaphylli, it is achieved
High-yield and high-efficiency is planted, and improves the health care of sugariness Herb Gynostemmae Pentaphylli simultaneously.
The present invention solves it and technical problem is that and realize by the following technical solutions.
A kind of implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, including:
Tissue culture: the outer implant of sugariness Herb Gynostemmae Pentaphylli be inoculated in culture medium, carries out tissue training
Support, it is thus achieved that tissue cultured seedling, tissue culture includes that induction of callus, differentiation are cultivated
And root culture；Three garden breedings are set up: tissue cultured seedling is transplanted in nursery build up strain nursery；Will
The rhizome of the Herb Gynostemmae Pentaphylli strain plant growing 1 year in strain nursery carries out plantation and builds up expanding propagation garden；Will
The rhizome of the Herb Gynostemmae Pentaphylli plant that expanding propagation garden grows 1 year carries out plantation and builds up garden, land for growing field crops kind；And
Cultivate administration: the rhizome of the Herb Gynostemmae Pentaphylli plant growing 1 year in garden, land for growing field crops kind is planted
Land for growing field crops.
The useful effect of the implantation methods of the organic selenium-rich sugariness Herb Gynostemmae Pentaphylli that the embodiment of the present invention provides
Fruit is: the present invention utilizes tissue culture technique to set up strain nursery, and tissue culture can keep sugariness
The kind of Herb Gynostemmae Pentaphylli, tissue culture can Fast-propagation sugariness Herb Gynostemmae Pentaphylli, substantially increase sweet
The yield of taste Herb Gynostemmae Pentaphylli, is obtained in that the tissue cultured seedling of virus-free simultaneously, then uses by tissue culture
This seedling is bred, then the sugariness Herb Gynostemmae Pentaphylli planted or would not seldom occur virosis.Have
It is beneficial to improve the quality of sugariness Herb Gynostemmae Pentaphylli, reduces planting cost simultaneously, then bred by three gardens
Foundation, make the tissue cultured seedling of tissue culture well be grown, by land for growing field crops strand in kind of garden
The rhizome of the blue plant of stock plants land for growing field crops, improves the quality and yield of sugariness Herb Gynostemmae Pentaphylli, it is achieved
High-yield and high-efficiency is planted.Nutrition through the sugariness Herb Gynostemmae Pentaphylli that the implantation methods of present invention offer is planted
Being worth height, health care is more excellent.
Detailed description of the invention
For making the purpose of the embodiment of the present invention, technical scheme and advantage clearer, below by right
Technical scheme in the embodiment of the present invention is clearly and completely described.In embodiment unreceipted
Actual conditions person, the condition advised according to normal condition or manufacturer is carried out.Agents useful for same or instrument
Device unreceipted production firm person, being can be by the commercially available conventional products bought and obtain.
Below the implantation methods of the organic selenium-rich sugariness Herb Gynostemmae Pentaphylli of the embodiment of the present invention is had
Body explanation.
The implantation methods of a kind of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, comprises the following steps:
S1, outer implant select: will put into screening culture medium through the to be selected outer implant disinfected
Middle cultivation filters out outer implant, and outer implant to be selected includes stem apex, stem section and blade；Screening and culturing
Base be in MS culture medium add 0.5～1.5mg/L 6-BA, 0.1～0.3mg/L NAA,
20～40g/L sucrose and 6～8g/L agar are made；Described outer implant is described stem apex.
Wherein, the manufacture method of screening culture medium is: first add suitable on the basis of MS culture medium
Amount water, in beaker, adds 6-BA 0.5～1.5mg, NAA 0.1～0.3mg, sucrose 20～40g
With agar 6～8g, then the graduated cylinder constant volume that adds water is to 1L, then debugs PH.
MS culture medium was Murashige and Skoog in 1962 is tobacco cell Training Design
, MS culture medium is currently used most common culture medium.Specifically, sweet in the present embodiment
The outer implant of taste Herb Gynostemmae Pentaphylli is selected from grace five leaf honey kind or the sweet tissue of grace seven leaf or organ.
Grace five leaf honey kind: utilize Herb Gynostemmae Pentaphylli mutation Gynostemma pentaphyllum
Makino DanTian No.9926835 is maternal, with Gynostemma pentaphyllum
Makino DanTian No.17892 is paternal hybrid, to its F1Individual selection is carried out for colony,
Choose the F that performance is excellent1Strain, uses its F1Strain Propagation of Rhizomes seedling, obtains " grace five
Leaf honey " new varieties, its Propagation of Rhizomes descendant inheritting is stable.
Grace seven leaf sweets kind: utilize Herb Gynostemmae Pentaphylli mutation Gynostemma pentaphyllum
Makino DanTian No.19371 is maternal and Gynostemma pentaphyllum
Makino DanTian No.592 is paternal hybrid, to its F1Individual selection is carried out for colony,
Choose the F that performance is excellent1Strain；Use its F1Strain Propagation of Rhizomes seedling, obtains " grace seven
Ye Tian " new varieties, its Propagation of Rhizomes descendant inheritting is stable.
Sugariness Herb Gynostemmae Pentaphylli new varieties eliminate the defect of bitterness Herb Gynostemmae Pentaphylli former mutation bitter in the mouth imapirment of the spleen and stomach,
Medicinal ingredient does not reduce compared with the former mutation of bitterness Herb Gynostemmae Pentaphylli, and not only ginsenoside is greatly improved, and
And sugariness Herb Gynostemmae Pentaphylli new varieties contain the unexistent medicinal ingredient of bitterness Herb Gynostemmae Pentaphylli, such as: peculiar
Flavone, aromatic substance, sapogenin, sugariness element etc., these peculiar medicinal ingredients have the highest medicine
With and health value.
Sweetleaf Herb Gynostemmae Pentaphylli new varieties medical value is high, and quality better, grace seven leaf is sweet containing gynostemma pentaphyllum total soap
Glycoside 5.52%, amino acid/11 4.2%, vitamin E 108.70mg/kg, than promote both at home and abroad
Former mutation respectively the highest by 33.3%, 11.8%, 66%；Per mu yield 290kg, more former than promote both at home and abroad
Mutation per mu yield 243kg height 47kg；Grace five leaf honey contains Herb Gynostemmae Pentaphylli total glycosides 5.56%, aminoacid
13.3%, vitamin E 6.72mg/kg, respectively higher by 34.3% than the former mutation promoted both at home and abroad,
4.7%, 17.2%；Per mu yield 274kg, the ratio former mutation high 31kg of per mu yield 243kg promoted both at home and abroad,
Improve its medical value, expand application approach.
So far, what Herb Gynostemmae Pentaphylli carried out isolated culture has 5 kinds of outer implant, including stem section,
Stem apex, blade, petiole, bud etc..Under suitable condition of culture, choose different sources,
Different cultivars, the tender stem segments of healthy and strong Herb Gynostemmae Pentaphylli plant of different growing stage, stem apex, blade,
Petiole, bud etc., as outer implant, are disinfected through suitable, are seeded in hormon proportioning
MS culture medium on, its callus induction, differentiation, root culture and be migrated to
Live and the most easily carry out, but result has different.
Research callus induction rate finds with outer implant selection dependency: the callus induction of blade
Rate is the highest, but the callus formed is yellow green, more fragile, is difficult to differentiate further not
Normal bud, is difficult to successive transfer culture；The callus of the stem section formation that children is tender is in green, loose, swollen
Greatly, inductivity is higher；Stem tip induction rate is relatively low, but the callus induced is more easy to survival,
Growth potential is good and easily differentiates adventitious bud, can avoid the endophyte shadow to whole fast numerous process
Ring.Therefore, the stem apex of robust growth is chosen as outer implant. it is to prevent having most of variety deterioration
Effect approach, is greatly improved reproduction speed with this regrowth cultivated, and suitable Clonal regeneration is advised
Modelling produces.
Meanwhile, a kind of side disinfected for sugariness Herb Gynostemmae Pentaphylli is embodiments provided
Method, after outer implant liquid detergent or detergent that concentration is 5% rinsing 3～8min, with stream
Water rinses 0.5～1h, and then by 75% soak with ethanol 25～35s, sterile deionized water rinses 4～6
Time, being subsequently placed into concentration is vibration sterilization 7～10min or with secondary chlorine in 0.1%HgCl2 solution
Acid sodium sterilization 22～27min, then rinses 5～6 times with sterile deionized water.
After washing 0.5～1h with flowing water, before by 75% soak with ethanol 25～35s,
Need to suck moisture unnecessary in outer implant with filter paper, and outer implant moved into superclean bench,
Carry out subsequent operation.
After outer implant has been disinfected, the external body after sterilization is placed in sterilizing culture dish
In, then outer implant cuts segment, and (length 0.5～1.5cm, with 1 axillary-bud or top-bud, excision
The wound that medicinal liquid contacted), then the outer implant after cutting is inoculated in screening culture medium.
S2, tissue culture: the outer implant of sugariness Herb Gynostemmae Pentaphylli is inoculated in culture medium, carries out group
Knitting cultivation, it is thus achieved that tissue cultured seedling, described tissue culture includes induction of callus, Multiple Buds
Differentiation culture and root culture.
Specifically, induction of callus includes outer implant is inoculated into callus induction training
Support in base, outer implant is formed callus.
Take 1.0cm stalwartness stem apex, to contain 3% sucrose and the MS of 0.7% agar by densitometer
Culture medium based on culture medium, the KT and 2 of interpolation variable concentrations, 4-D, 6-BA and NAA,
Tri-kinds of hormone combinations of 6-BA and 2,4-D, observe the induction to callus and the shadow of growth potential
Ring.
Result shows, KT and 2,4-D combination is relatively low to inductivity and the survival rate of callus,
Growing way is slower；Induction is combined by the 1.5mg/L 6-BA and 0.3mg/L NAA of densitometer
Callus is that peak green, structure are relatively tight, growth potential is preferable, and inductivity reaches 92.6%；Press
The callus of 1.0mg/L 6-BA and the 2.0mg/L 2,4-D combination induction of densitometer is yellow
White, loosen, expand, water content higher, inductivity reaches 91.9%.
It is therefore preferred that the optimal hormone combinations of callus induction is 1.5mg/L 6-BA
And 0.3mg/LNAA.Callus inducing medium is to add in MS culture medium
1～2mg/L 6-BA, 0.2～0.4mg/L NAA, 20～40g/L sucrose and 6～8g/L agar systems
?.
Specifically, differentiation is cultivated and is included being transferred in division culture medium callus,
And differentiate Multiple Buds.
Stem apex callus is inoculated on division culture medium through 2-3 successive transfer culture, the
6d starts Multiple Buds occur, and after 25d, the inductivity of Multiple Buds is up to 93%.Add merely growth
Element (NAA) is conducive to induction and the formation of root of callus, adds merely the basic element of cell division
(6-BA) also can callus induction, and differentiate Multiple Buds, and two classes adding low concentration be raw
Long element (NAA and 2,4-D) and the basic element of cell division (6-BA) of high concentration, the differentiation to Multiple Buds
Frequency is apparently higher than the combination the most individually adding the basic element of cell division.
Preferably, division culture medium be in MS culture medium add 0.1～0.3mg/L NAA,
0.1～0.3mg/L 2,4-D, 0.5～1.5mg/L 6-BA, 20～40g/L sucrose and 6～8g/L fine jades
Fat.
Specifically, root culture includes being transferred in root media Multiple Buds, grows root,
Form tissue cultured seedling.
When sprout length is to about about 2cm, cuts single sprout and proceed to 1/2MS culture medium is entered
Row root culture.Result shows, can take root when NAA is used alone, and with concentration is
1.5mg/L's is optimal, but adds paclobutrazol (PP in the medium333After) can hestening rooting,
PP333Best results when concentration is 0.3mg/L；After adding root-inducing powder, root system is many, and root is sturdy,
Sprout is sturdy, and leaf color is dark green, and root-inducing powder is optimal with 100mg/L concentration.After considering,
Root media is addition 1～2mg/L NAA, 0.2～0..4mg/L in 1/2MS culture medium
PP333Make with 90～110mg/L root-inducing powders.
Specifically, plant tissue culture is had by different illumination intensity, periodicity of illumination and temperature conditions
Appreciable impact.Temperature is higher than 24～26 DEG C, and tissue cultured seedling growth is thin and delicate, the most sturdy；Light application time
Higher than 11～13h/d, tissue cultured seedling Failure to thrive is contained, and light application time is less than 11～13h/d, tissue cultured seedling
Grow thin and delicate；Intensity of illumination is higher than 1500～2000lx, and tissue cultured seedling turns to be yellow, and intensity of illumination is less than
1500～2000lx tissue cultured seedling poor growths, prolongation group training time more than 15d, and tissue cultured seedling
Thin and delicate, short and small.
It is therefore preferable that the outer induction of implant callus of sugariness Herb Gynostemmae Pentaphylli, differentiation and life
Root cultivate temperature be 24～26 DEG C, light application time 11～13h/d, intensity of illumination be
Carry out under conditions of 1500～2000lx.
After under the conditions of basis, callus inducing medium cultivates 7d, outer implant otch base portion is dashed forward
Rising and expand, the periphery of otch starts wound healing, and whole wound healing after 2 weeks, callus lures
Conductance average out to more than 95%, and after 1 week, just can induce substantial amounts of aseptic adventitious bud, 3
A large amount of crowd shoots can be bred after week；Crowd shoots is proceeded to about 15d after root media
Left and right sends out roots.
Additionally, the acid-base value of culture medium is also studied by the embodiment of the present invention, culture medium
The growth of the external implant of acid-base value does not directly affect, but there are certain remote-effects, properly
PH value be not only advantageous for outer planting bulk-growth, and contribute to culture medium solidifying, work as pH value
During higher than 6.5, culture medium can be hardening；During less than 5.0, agar can not solidify well.Cause
This, in the embodiment of the present invention, the pH value of preferred culture medium is 5.8～6.4.
In order to improve the tissue cultured seedling adaptive faculty to land poor environment, shorten transplanting seedling time, strengthen
The tissue cultured seedling resistivity to low temperature, strong wind etc..Tissue cultured seedling is being transplanted to by the embodiment of the present invention
Before nursery, first tissue cultured seedling is carried out seedling exercising.
Specifically, when tissue cultured seedling take root number reach about 10, height of seedling about 5cm time, permissible
Carry out seedling exercising: bottle cap or sealed membrane are opened under shading screen, add in media surface in good time
The sterilized water of thin layer, removes Seedling after adapting to 5d, is transplanted to after carefully washing most residual media
Cultivate equipped with in the nutritive cube of Nutrition Soil.Culturing pot is placed in shading shed, keeps temperature 15 DEG C
Above, humidity about 90%, survival rate averagely reaches 98%, and after cultivation, 7d restore normal growth,
After 15d, plant reaches about 10cm, and after 1 month, plant height is up to 20cm.Now can be by nutrition
Daily management is carried out, it is possible to directly transplant on big Tanaka, side during Seedling moves to other cool canopy in alms bowl
The little tree branch inserting band leaf shades.The transplanting survival rate of Herb Gynostemmae Pentaphylli tissue cultured seedling is higher, typically up to
More than 95%.
When tissue cultured seedling take root number more than 10, height of seedling 6cm time seedling exercising, although its survival rate is also
Reaching 98%, but because culture medium nutrition tails off, incubation time extends to 6 days, plant volume becomes
Greatly, light and condition are deteriorated so that it is tissue cultured seedling plant dies down, unfavorable acclimatization and transplants, extends simultaneously
The group training time, also can increase group training light at cost.
Height of seedling less than 5cm time, culture medium nutrition does not make full use of, and Seedling is too small, seedling exercising and
Field-transplanting survival rate is less than 98%, and survival rate is in the range of 80%-91%.
It is preferred, therefore, that when tissue cultured seedling take root number reach about 10, height of seedling about 5cm time, can
To carry out seedling exercising.
S3, three garden breedings are set up: be transplanted in nursery build up strain by the tissue cultured seedling after seedling exercising
It it is garden；The rhizome of the Herb Gynostemmae Pentaphylli strain plant growing 1 year in strain nursery is carried out plantation and builds up expansion
Numerous garden；The rhizome of the Herb Gynostemmae Pentaphylli plant that expanding propagation garden is grown 1 year carries out plantation and builds up land for growing field crops kind
Garden.
Specifically, sugariness Herb Gynostemmae Pentaphylli tissue cultured seedling is transplanted to nursery after seedling exercising in seedling exercising canopy.Seedling
Garden substrate base fertilizer mixes decomposed manure 3000kg/ mu, Se-enriched bio-organic fertilizer 80kg/ mu, is combined
Microbial bacterial agent fertilizer 4kg/ mu, railway carriage or compartment width 1.2m, seeding row spacing 15cm × 15cm, railway carriage or compartment furrow width 0.3m,
Every 1, cave, plants into 1 railway carriage or compartment each elected strain seed, builds up strain nursery.
Select the rhizome of the Herb Gynostemmae Pentaphylli strain plant that the growth stalwartness of 1 year is anosis in strain nursery, use
The agricultural streptomycin infiltration 0.4～0.6min diluting 1800～2200 times is pulled out, by 3-4 stipes
It is 1 section, is truncated into kind of a root, plant by seeding row spacing 15cm × 15cm kind, 1, every cave, build up expansion
Numerous garden.
By Herb Gynostemmae Pentaphylli plant rhizome anosis for the expanding propagation garden growth stalwartness of 1 year, with dilution
The agricultural streptomycin infiltration 0.4～0.6min of 1800～2200 times is pulled out, is 1 by 3-4 stipes
Section, is truncated into kind of a root, plants by seeding row spacing 15cm × 15cm kind, 1, every cave, builds up land for growing field crops and uses
Plant garden, plant 1 year for Production of Large Fields kind.
S4, Cultivate administration: by land for growing field crops rhizome kind of the growth Herb Gynostemmae Pentaphylli plant of 1 year in kind of garden
Plant to land for growing field crops.
Selection of land: the rhizome of the Herb Gynostemmae Pentaphylli plant growing 1 year in garden, land for growing field crops kind is planted
Before, first select arable field, preferably field to be positioned at height above sea level 400-1600m, have
The region of irrigation condition, near have methane-generating pit, have pouring root and equipment of sprinkler irrigation, available immersible pump is even
Biogas manure is delivered to field fertilization by adapter road.Soil is fertile soil, sandy loam or loam,
PH6.0-7.5, previous crops can not be solanaceous crops or the ground family crops such as Rhizoma Solani tuber osi, Nicotiana tabacum L., Fructus Lycopersici esculenti,
The highest temperature is less than 30 DEG C, and the lowest temperature is not less than-9.5 DEG C.
Whole field: turn over before strain nursery, expanding propagation garden, land for growing field crops kind of garden, field seeding dry field once,
Whole fine earth earth, flattens field, makees railway carriage or compartment 1.2m width, railway carriage or compartment furrow width 0.3m, ditch depth 0.25m.
The soil of Herb Gynostemmae Pentaphylli continuous cropping, after annual results, field is poured water and is flooded field soil 7d, kills
Nematicide in field, eliminates Herb Gynostemmae Pentaphylli rhizosphere insect；If building Herb Gynostemmae Pentaphylli planting matrix in dry land region
Ground, then before plantation Herb Gynostemmae Pentaphylli, 15d combines whole field and uses biocontrol bacterial strain SR18 nematode killer
(effectively live spore count >=10,000,000,000/g), mu 160g mixes fine earth mixing and is spread on planting pit
In or field planting ditch in.
Fertilising: strain nursery, expanding propagation garden, land for growing field crops kind garden, every mu of the base fertilizer in land for growing field crops use base fertilizer:
Decomposed manure 4500～5500kg, complex micro organism fungicide fertile 1～3kg and Se-enriched bio are organic
Fertile 70～90kg.
Wherein, decomposed manure is that people's Animal fecal pollution is through EM (effective microorganisms fermentation skill
Art) method, hot fermentation heap corruption method, the self-control fermentation Harmless such as heap corruption method that accelerates the ripening processes, kills
Gone out various parasitic ovum, pathogen, weed seed, and eliminate harmful organic acid and have
The fertilizer of evil gas.
Complex micro organism fungicide is by the microorganism fungus kind system of two or more mutual the most not antagonism
The microorganism formulation become.
Se-enriched bio-organic fertilizer is that multiple beneficial microorganism species is combined formation with selenium-enriched organic fertilizer
Novel, efficient, safe microorganism-fertilizer compound fertilizer.
Decomposed manure, complex micro organism fungicide and the Se-enriched bio used in the embodiment of the present invention
Fertilizer all can directly purchased on the market.
Plantation: after applying fertilizer field, the Herb Gynostemmae Pentaphylli growing 1 year in selecting garden, land for growing field crops kind is planted
The grace five leaf honey of the Propagation of Rhizomes rejuvenation of strain, the plantation of grace seven leaf sweet detoxic seedling, 3-5 stipes
One, the every cave of Herb Gynostemmae Pentaphylli rhizome, plants specification 15cm × 15cm, planting depth 5cm.Plantation
Rear covering black mulch film, plays insulation and the effect of controlling weeds growth, is stabbed by mulch film after emerging
Broken, allow Herb Gynostemmae Pentaphylli Seedling expose mulch film.
Cropping pattern can use three-dimensional built, floor file etc. according to different altitude height, terrain
Multiple cultivation mode, during corn relay interplanting, corn seeding is in the central authorities in face, railway carriage or compartment, the double strain in every cave.
Weeding: Feng Hangqian uproots clean weeds, sees that grass i.e. removes later.
The field envelope departure date: imposing biogas manure 20 when emerging and carry on a shoulder pole/mu, Herb Gynostemmae Pentaphylli height of seedling 30cm takes advantage of
Fine day is pinched, and often plucks 1 time and sprays fertilizing of biogas manure 1 time, each 20 loads/mu.
Topdress: use complex micro organism fungicide in peak of growing season fertile, use richness in the stem and leaf collecting period
Organic selenium liquid fertilizer, imposes biogas manure 20 after plucking stem and leaf and carries on a shoulder pole/mu.Complex micro organism fungicide
Fertilizer is used: with the fertile aqueous solution to dilute 250 times of complex micro organism fungicide in peak of growing season pouring root
1 time, consumption is 50kg/ mu, later fertile to dilute the water of 500 times with complex micro organism fungicide
Solution interval 30d sprays 1 time again, and consumption is 50kg/ mu, twice totally.Composite microbial bacteria
Sugariness Herb Gynostemmae Pentaphylli can be played the effect of diseases prevention by agent fertilizer.
Organic selenium-rich liquid fertilizer is used: in the stem and leaf collecting period, plucks outer every mu of 15d root before stem and leaf
Spraying 16kg selenium content is 0.30mg/ml organic selenium-rich liquid fertilizer, and interval 30d sprays again later
Execute 1 time, twice totally；Impose biogas manure 20 after plucking stem and leaf every time and carry on a shoulder pole/mu.Organic selenium-rich liquid
State fertilizer can be soil selenium supplement, and biogas manure can supplement the nutrients for soil, makes sugariness Herb Gynostemmae Pentaphylli
Selenium content higher, nutrition is more abundant.
Wherein, biogas manure and complex micro organism fungicide fertilizer can the most directly purchased.
And organic selenium-rich liquid fertilizer is by bacillus subtilis is inoculated into test tube rejuvenation cultivation
Cultivating in base and form test tube kind, then test tube kind being inoculated in fermentation tank carries out fermenting prepares.
Wherein, the composition of test tube rejuvenation culture medium includes: 6g/L glucose, 5g/L sucrose,
5g/L Carnis Bovis seu Bubali cream, 5g/L peptone, 4g/L sodium chloride, 18g/L agar；In described fermentation tank
Including: 6g/L glucose, 5g/L sucrose, 5g/L Carnis Bovis seu Bubali cream, 10g/L peptone, 4g/L
Sodium chloride, biofermentation defoamer 1g/L.
Wherein in test tube rejuvenation culture medium and fermentation tank, the concentration of material refers to that material is in 1L water
Content.
Specifically, when preparing 1 ton of organic selenium-rich liquid fertilizer containing organic selenium 300mg/L,
(1) test tube rejuvenation culture medium composition: 6g glucose, 5g sucrose, 5g Carnis Bovis seu Bubali cream,
10 peptones, 4g sodium chloride, 18g agar, water 1000mL, pH7.6；
(2) rejuvenation culture medium sterilizing: put radiated by gamma-ray machine 2.5kGy sterilizing, is put into tiltedly
Face, cooled and solidified；
(3) the bacillus subtilis test tube bacterium in 4 DEG C of cold preservations of refrigerator will take 1 with inoculation ring species
Ring strain is seeded in test tube rejuvenation culture medium, inoculates 100 test tube kinds and puts 32 DEG C of incubators
Middle cultivation covers with bacterium to SE201412 test tube slant；
(4) bacillus subtilis seed liquor amplification culture: take the tap water 100L dress of aeration
Enter in 200L fermentation tank, add 600g glucose, 500g sucrose, 500g Carnis Bovis seu Bubali cream,
1000g peptone, 400g sodium chloride, biofermentation defoamer 0.1kg, adjust with sodium hydroxide
PH7.6, with radiated by gamma-ray machine 3kGy disinfection fermentation tank breather line, disinfection fermentation tank
Tank body, sterilization fermentation liquid, then by 100 bacillus subtilis SE201412 examinations of inoculation
The inoculation of pipe kind accesses in fermentation tank, set fermentation mixing speed 195r/min, temperature in tank
32 DEG C, pressure inside the tank 0.05MPa, in tank culture fluid volume be passed through volume of air ratio 1: 0.3
(V/V).Fermentor cultivation 15h, after accessing seed, samples fermentation liquid acidity every 3h
Meter measures pH (keeping fermentation tank fermentation liquid during the fermentation to keep pH7.6) and with 722
Visible spectrophotometer measures bacterial cell concentration, when finding that fermentation liquid 12h bacterial cell is dense
Degree and 15h bacterial cell concentration detect at maximum absorbance, when not being significantly increased, terminate
Fermentation.
(5) 1 tons of productions containing organic selenium 300mg/L organic selenium-rich liquid fertilizer: at 2 tons
In fermentation tank add 1 ton of aeration tap water, add 6000g glucose, 5000g sucrose,
5000g Carnis Bovis seu Bubali cream, 10000g peptone, 4000g sodium chloride, purity 99% sodium selenite
673.7g, GPE-6330 biofermentation special defoamer 1kg, adjusts pH7.6, uses gamma ray
Irradiator 4kGy sterilizing, fermentation tank breather line, sterilization fermentation tank, sterilization fermentation liquid,
Then 100 bacillus subtilis SE201412 test tube kind inoculations of inoculation will access fermentation
In tank, set fermentation mixing speed 195r/min, temperature 32 DEG C, pressure inside the tank in tank
0.05MPa, in tank culture fluid volume be passed through volume of air than 1: 0.3 (V/V).Fermentation tank
Cultivate 15h, after accessing seed, measure pH every 3h sampling fermentation liquid acidometer and (keep
Fermentation tank fermentation liquid during the fermentation keeps pH7.6) and use 722 visible spectrophotometers
Measure bacterial cell concentration, when finding fermentation liquid 18h bacterial cell concentration and 15h bacterial cell
Concentration detects at maximum absorbance, when not being significantly increased, terminates fermentation, pipeline input storage
Tank, uses automatic filling machine fill, and every bottle fills 5L, with radiated by gamma-ray machine 3kGy after capping
Can deposit after sterilizing 1 year.
Certainly, the organic selenium-rich liquid fertilizer that the embodiment of the present invention provides can enter according to specific needs
Row equal proportion scaling preparation, however it is not limited to 1 ton provided in the embodiment of the present invention is containing organic selenium
The preparation of the organic selenium-rich liquid fertilizer of 300mg/L.
Below three-dimensional planting mode, floor file cultivation mode, corn relay interplanting planting pattern are had
Body illustrates:
Three-dimensional planting mode:
Three-dimensional planting mode district tries and demonstration effect is: plant what 25000 strain Herb Gynostemmae Pentaphylli processed
Herb Gynostemmae Pentaphylli produces fresh leaf 7300kg/667m2；Plant 20000 strain Herb Gynostemmae Pentaphylli, produce fresh leaf 5695
kg/667m2；Plant 15000 strain Herb Gynostemmae Pentaphylli and produce fresh leaf 456.05kg/667m2；Variance analysis,
Solid cultivation increases Herb Gynostemmae Pentaphylli strain number, and the volume increase of Herb Gynostemmae Pentaphylli fresh leaf is the most notable.
Floor file cultivation mode:
Floor file cultivation area is tried and demonstration effect is: plants 15000 strain Herb Gynostemmae Pentaphylli and all produces fresh leaf
2265.5kg/667m2；Plant 20000 strain Herb Gynostemmae Pentaphylli and all produce fresh leaf 2905.5kg/667m2；Kind
Plant 25000 strain Herb Gynostemmae Pentaphylli and all produce fresh leaf 3433.5kg/667m2；Variance analysis, increases strand
Blue strain strain number, Herb Gynostemmae Pentaphylli fresh leaf increase yield significantly.
Semen Maydis intercropping and interplanting cropping pattern:
Try and cultivation mode and yield such as following table in demonstration with corn relay interplanting district:
The results of analysis of variance shows, Semen Maydis 3500 strain, and Herb Gynostemmae Pentaphylli 15000 strain processes, Semen Maydis
Volume increase is extremely notable, and Herb Gynostemmae Pentaphylli volume increase is the most notable.
With sugariness Herb Gynostemmae Pentaphylli and corn intercrop, soil out of question, growing space out of question, illumination out of question,
Corn yield is suitable with nonoculture, and Herb Gynostemmae Pentaphylli per mu yield fresh leaf 1130kg-1315 produces blade tea product and increases
Receive 2000 yuan-3000 yuan/667m2, in sugariness producing region, the Herb Gynostemmae Pentaphylli that every mu of family of plantation is plucked is fresh
Leaf and the young stem and leaf output value, more than 15000 yuan, it is achieved that grain money gets bumper harvests, and greatly improves
Land utilization ratio and raising unit are economic benefit.
Carry out sweet taste Herb Gynostemmae Pentaphylli high-yield and high-efficiency production model research in planted in different ecological areas, filter out sweet
The nonoculture patterns such as taste Herb Gynostemmae Pentaphylli solid built, floor file and with high stalk crops, such as: Semen Maydis,
Multiple cultivation mode plantation with the intercropping pattern such as economic forest.Can enter according to actual situation
Row selects.
The present invention utilizes tissue culture technique to set up strain nursery, and tissue culture can keep sugariness to twist
The kind that stock is blue, tissue culture can Fast-propagation sugariness Herb Gynostemmae Pentaphylli, substantially increase sugariness
The yield of Herb Gynostemmae Pentaphylli, is obtained in that the tissue cultured seedling of virus-free simultaneously, then uses this by tissue culture
Seedling is bred, then the sugariness Herb Gynostemmae Pentaphylli planted or would not seldom occur virosis.Favorably
In the quality of raising sugariness Herb Gynostemmae Pentaphylli, reduce planting cost, then by three gardens breedings simultaneously
Set up, make the tissue cultured seedling of tissue culture well be grown, by land for growing field crops strand in kind of garden
The rhizome of blue plant plants land for growing field crops, rich at land for growing field crops bottom application Se-enriched bio-organic fertilizer and root outer spraying
Organic selenium microbial bacterial agent.Improve the quality and yield of sugariness Herb Gynostemmae Pentaphylli, it is achieved high-yield and high-efficiency
Plantation.Through being of high nutritive value of sugariness Herb Gynostemmae Pentaphylli of the implantation methods plantation that the present invention provides,
Health care is more excellent.And the total Se content of sugariness Herb Gynostemmae Pentaphylli stem and leaf reaches 0.2-5.0mg/kg, organic selenium
Mass fraction >=80%, reaches DBS42/002-2014 " requirement of organic selenium-rich food Se content "
Index.
Embodiment one
A kind of implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, including:
Tissue culture: temperature be 24 DEG C, light application time 11h/d, intensity of illumination be 1500lx
Under conditions of carry out.The outer implant of sugariness Herb Gynostemmae Pentaphylli is inoculated into callus inducing medium
In, outer implant is formed callus, then callus is transferred in division culture medium,
And differentiate Multiple Buds, when the sprout length of Multiple Buds is to about about 2cm, cut single bud
Seedling is transferred in root media, grows root, forms tissue cultured seedling.
Wherein: callus inducing medium by densitometer consist of containing 1mg/L 6-BA,
0.2mg/L NAA, 20g/L sucrose and the MS culture medium of 6g/L agar.
Division culture medium by densitometer consist of containing 0.2mg/L NAA, 0.2mg/L 2,4-D,
0.5mg/L 6-BA, 20g/L sucrose and the MS culture medium of 6g/L agar.
Root media consisting of containing 1mg/L NAA, 0.2mg/L PP by densitometer333With
The 1/2MS culture medium of 90mg/L root-inducing powder.
Three garden breedings are set up: decomposed manure 3000kg/ mu mixed by nursery substrate base fertilizer, and selenium-rich is raw
Organic fertilizer 80kg/ mu, complex micro organism fungicide fertilizer 4kg/ mu, railway carriage or compartment width 1.2m, seeding row spacing 15cm
× 15cm, railway carriage or compartment furrow width 0.3m, 1, every cave, each elected strain seed is planted into 1 railway carriage or compartment,
Build up strain nursery.
Select the rhizome of the Herb Gynostemmae Pentaphylli strain plant that the growth stalwartness of 1 year is anosis in strain nursery, use
Dilute 1800 times agricultural streptomycin infiltration 0.4min pull out, be 1 section by 3-4 stipes,
It is truncated into kind of a root, plants by seeding row spacing 15cm × 15cm kind, 1, every cave, build up expanding propagation garden.
By Herb Gynostemmae Pentaphylli plant rhizome anosis for the expanding propagation garden growth stalwartness of 1 year, with dilution 1800
Agricultural streptomycin infiltration 0.4min again pulls out, is 1 section by 3-4 stipes, is truncated into kind of a root,
Plant by seeding row spacing 15cm × 15cm kind, 1, every cave, build up garden, land for growing field crops kind, plant 1 year
For Production of Large Fields kind.
Cultivate administration: select suitable land for growing field crops, at Shang Zuo railway carriage or compartment, land for growing field crops 1.2m width, railway carriage or compartment furrow width 0.3
M, ditch depth 0.25m.Then land for growing field crops being carried out parasite killing and fertilising, every mu of the base fertilizer in land for growing field crops is used
Decomposed manure 4500kg, complex micro organism fungicide fertilizer 1kg and Se-enriched bio-organic fertilizer 70kg.
The most again the rhizome of the Herb Gynostemmae Pentaphylli plant growing 1 year in garden, land for growing field crops kind is planted to land for growing field crops.This
Embodiment uses three-dimensional planting mode, one, the every cave of Herb Gynostemmae Pentaphylli rhizome of 3 stipes, plantation
Specification is 15cm × 15cm, planting depth 5cm.Cover black mulch film after plantation, wait to emerge
After mulch film is poked, allow Herb Gynostemmae Pentaphylli Seedling expose mulch film.Then Gynostemma pentaphyllum Makino is managed, and first uproots
Clean weeds, then impose biogas manure 20 and carry on a shoulder pole/mu when emerging, and Herb Gynostemmae Pentaphylli height of seedling 30cm takes advantage of fine
It is pinched, and often plucks and sprays biogas slurry 1 time 1 time, each 20 loads/mu.Then composite microbial is used
The fertile aqueous solution with dilution 250 times of thing microbial inoculum is in peak of growing season pouring root 1 time, and consumption is 50kg/
Mu, sprays with the fertile aqueous solution interval 30d with dilution 500 times of complex micro organism fungicide later again
Executing 1 time, consumption is 50kg/ mu, twice totally.In the stem and leaf collecting period, pluck 15d before stem and leaf
Outside root, every mu sprays 16kg selenium content is 0.30mg/ml organic selenium-rich liquid fertilizer, is spaced later
30d sprays 1 time again, twice totally；Impose biogas manure 20 after plucking stem and leaf every time and carry on a shoulder pole/mu.
Embodiment two
A kind of implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, including:
Outer implant selects: in the present embodiment, before tissue culture procedures, also include outer planting
Body selects: the to be selected outer implant through disinfecting is put into cultivation in screening culture medium and filters out
Outer implant, outer implant to be selected includes stem apex, stem section and blade；Outer implant is stem apex.
Wherein, screening culture medium consisting of containing 1mg/L6-BA, 0.1mg/L by densitometer
The MS culture medium of NAA, 30g/L sucrose and 7g/L agar.
Sterilization is by after outer implant liquid detergent or detergent that concentration is 5% rinsing 5min,
Rinsing 1h with flowing water, then by 75% soak with ethanol 30s, sterile deionized water rinses 5 times,
Be subsequently placed into concentration be in 0.1%HgCl2 solution vibration sterilization 8min, then with aseptic go from
Sub-water rinses 5 times.
After washing 1h with flowing water, before by 75% soak with ethanol 30s, need to use
Filter paper sucks moisture unnecessary in outer implant, and outer implant is moved into superclean bench, after carrying out
Continuous operation.
After outer implant has been disinfected, the external body after sterilization is placed in sterilizing culture dish
In, then outer implant is cut segment, then the outer implant after cutting is inoculated into screening culture medium
In.
Tissue culture: the tissue culture of the present embodiment is to be 25 DEG C, light application time in temperature
12h/d, intensity of illumination are to carry out under conditions of 1800lx.
Wherein: screening culture medium consisting of containing 1mg/L 6-BA, 0.1mg/L by densitometer
The MS culture medium of NAA, 30g/L sucrose and 7g/L；
Callus inducing medium by densitometer consist of containing 1.5mg/L 6-BA,
0.3mg/L NAA, 30g/L sucrose and the MS culture medium of 7g/L.
Division culture medium by densitometer consist of containing 0.1mg/L NAA, 0.1mg/L 2,4-D,
1mg/L 6-BA, 30g/L sucrose and the MS culture medium of 7g/L.
Root media consisting of containing 1.5mg/L NAA, 0.3mg/L PP by densitometer333
1/2MS culture medium with 100mg/L root-inducing powder.
Three garden breedings are set up: three provided with embodiment one are set up in three garden breedings in the present embodiment
Garden breeding establishment step is essentially identical, and difference is:
Preferably in the Herb Gynostemmae Pentaphylli strain that the stalwartness that will grow 1 year in strain nursery is anosis in the present embodiment
It is that the Herb Gynostemmae Pentaphylli plant rhizome growing the stalwartness of 1 year in the rhizome of plant and expanding propagation garden anosis enters
Before row plantation, first with the agricultural streptomycin infiltration of dilution 2000 times, and all at 0.5min
After pull out.
Cultivate administration: the Cultivate administration step that the Cultivate administration in the present embodiment and embodiment one provide
The most essentially identical, difference is:
The present embodiment use floor file cultivation mode to land for growing field crops growth strand of 1 year in kind of garden
The rhizome of blue plant is planted, and every mu of the base fertilizer in land for growing field crops use decomposed manure 5000kg,
Complex micro organism fungicide fertilizer 2kg and Se-enriched bio-organic fertilizer 80kg.
Embodiment three
A kind of implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, its step includes:
Outer implant selects: the outer implant provided in the present embodiment selects step to carry in embodiment two
Confession essentially identical, difference is: carry out the screening training of outer implant selection in the present embodiment
Support base consisting of containing 1.5mg/L 6-BA, 0.3mg/L NAA, 40g/L sucrose by densitometer
MS culture medium with 8g/L agar.
Tissue culture: the tissue culture of the present embodiment is to be 26 DEG C, light application time in temperature
13h/d, intensity of illumination are to carry out under conditions of 2000lx.
Callus inducing medium consisting of containing 2mg/L 6-BA, 0.4mg/L by densitometer
The MS culture medium of NAA, 40g/L sucrose and 8g/L agar.
Division culture medium by densitometer consist of containing 0.3mg/L NAA, 0.3mg/L 2,4-D,
1.5mg/L 6-BA, 40g/L sucrose and the MS culture medium of 8g/L agar.
Root media consisting of containing 2mg/L NAA, 0.4mg/L PP by densitometer333With
The 1/2MS culture medium of 110mg/L root-inducing powder.
Seedling exercising: tissue cultured seedling, before tissue cultured seedling is transplanted to nursery, is first entered by the embodiment of the present invention
Row seedling exercising.When tissue cultured seedling take root number reach 10, height of seedling 5cm time, carry out seedling exercising: shading
Off the net bottle cap or sealed membrane are opened, add the sterilized water of thin layer in good time in media surface, suitable
After answering 5d, Seedling is removed, after carefully washing most residual media, be transplanted to the nutrition equipped with Nutrition Soil
Alms bowl is cultivated.Culturing pot is placed in shading shed, keeps temperature more than 15 DEG C, and humidity 90% is left
The right side, survival rate averagely reaches 98%, and after cultivation, 7d restore normal growth, and after 15d, plant reaches 10cm
Left and right, after 1 month, plant height is up to 20cm.
Three garden breedings are set up: be transplanted in nursery build up strain by the tissue cultured seedling after seedling exercising
Garden；The rhizome of the Herb Gynostemmae Pentaphylli strain plant growing 1 year in strain nursery is carried out plantation and builds up expanding propagation
Garden；The rhizome of the Herb Gynostemmae Pentaphylli plant that expanding propagation garden is grown 1 year carries out plantation and builds up land for growing field crops kind
Garden.
Three garden breedings in the present embodiment are set up the three garden breedings provided with embodiment one and are set up step
The most essentially identical, difference is:
Preferably in the Herb Gynostemmae Pentaphylli strain that the stalwartness that will grow 1 year in strain nursery is anosis in the present embodiment
It is that the Herb Gynostemmae Pentaphylli plant rhizome growing the stalwartness of 1 year in the rhizome of plant and expanding propagation garden anosis enters
Before row plantation, first with the agricultural streptomycin infiltration of dilution 2200 times, and all at 0.6min
After pull out.
Cultivate administration: the Cultivate administration step that the Cultivate administration in the present embodiment and embodiment one provide
The most essentially identical, difference is:
The present embodiment use Semen Maydis intercropping and interplanting cropping pattern land for growing field crops is grown 1 year in kind of garden
The rhizome of Herb Gynostemmae Pentaphylli plant plant, and decomposed manure used by every mu of the base fertilizer in land for growing field crops
5500kg, complex micro organism fungicide fertilizer 3kg and Se-enriched bio-organic fertilizer 90kg.
Implantation methods plantation system through the organic selenium-rich sugariness Herb Gynostemmae Pentaphylli that the embodiment of the present invention provides
Sugariness Herb Gynostemmae Pentaphylli be analyzed, including to the analysis of effective component of sugariness Herb Gynostemmae Pentaphylli, sugariness
Herb Gynostemmae Pentaphylli full stem and leaf sanitary index is analyzed and always selenium and organic selenium content analysis, and concrete data are joined
It is shown in Table 1, table 2 and table 3.
The effective ingredient of table 1. sugariness Herb Gynostemmae Pentaphylli；
Table 2. sugariness Herb Gynostemmae Pentaphylli full stem and leaf sanitary index；
Table 3. sugariness Herb Gynostemmae Pentaphylli full stem and leaf Se content
(1) analysis of effective component of sugariness Herb Gynostemmae Pentaphylli
According to the testing result at Ministry of Agriculture's food quality supervision verification test center (Wuhan), sweet
The effective ingredient of taste Herb Gynostemmae Pentaphylli is as follows:
The effective ingredient of table 1. sugariness Herb Gynostemmae Pentaphylli
In terms of result above: ginsenoside, dimension " grace seven leaf is sweet ", " grace five leaf honey " are raw
Element B1, vitamin E and total amino acid content are above former mutation.
(2) sugariness Herb Gynostemmae Pentaphylli full stem and leaf sanitary index is analyzed
According to the testing result at Ministry of Agriculture's food quality supervision verification test center (Wuhan), sweet
Taste Herb Gynostemmae Pentaphylli full stem and leaf sanitary index is as follows:
Table 2. sugariness Herb Gynostemmae Pentaphylli full stem and leaf sanitary index
Sugariness Herb Gynostemmae Pentaphylli full stem and leaf testing result meets the index of correlation of green product.
(3) sugariness Herb Gynostemmae Pentaphylli full stem and leaf Selenium Contents
According to the testing result of country's selenium-rich product quality supervision and inspection center (bestowing favour), sugariness
The total selenium of the full stem and leaf of Herb Gynostemmae Pentaphylli and organic selenium content are as follows:
Table 3 sugariness Herb Gynostemmae Pentaphylli full stem and leaf Se content
The sugariness Herb Gynostemmae Pentaphylli full stem and leaf total selenium of testing result and organic selenium content meet
The index of DBS42/002-2014 " requirement of organic selenium-rich food Se content ".
Embodiments described above is a part of embodiment of the present invention rather than whole enforcement
Example.The detailed description of embodiments of the invention is not intended to limit the model of claimed invention
Enclose, but be merely representative of the selected embodiment of the present invention.Based on the embodiment in the present invention, this
It is every other that field those of ordinary skill is obtained under not making creative work premise
Embodiment, broadly falls into the scope of protection of the invention.
Claims (10)
1. the implantation methods of an organic selenium-rich sugariness Herb Gynostemmae Pentaphylli, it is characterised in that including:
Tissue culture: the outer implant of sugariness Herb Gynostemmae Pentaphylli be inoculated in culture medium, carries out tissue training
Support, it is thus achieved that tissue cultured seedling, described tissue culture includes induction of callus, differentiation
Cultivate and root culture；
Three garden breedings are set up: be transplanted in nursery build up strain nursery by described tissue cultured seedling；By described
The rhizome of the Herb Gynostemmae Pentaphylli strain plant growing 1 year in strain nursery carries out plantation and builds up expanding propagation garden；Will
The rhizome of the Herb Gynostemmae Pentaphylli plant that described expanding propagation garden grows 1 year carries out plantation and builds up garden, land for growing field crops kind；
And
Cultivate administration: by the rhizome kind of the growth Herb Gynostemmae Pentaphylli plant of 1 year in garden, described land for growing field crops kind
Plant to land for growing field crops.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 1,
It is characterized in that, before being additionally included in described tissue culture, carry out outer implant selection, will be through disappearing
The to be selected outer implant that poison processes is put into cultivate in screening culture medium and is filtered out described outer implant, described
Outer implant to be selected includes stem apex, stem section and blade；Described screening culture medium is in MS culture medium
Middle addition 0.5～1.5mg/L 6-BA, 0.1～0.3mg/L NAA, 20～40g/L sucrose and
6～8g/L agar are made；Described outer implant is described stem apex, and described sugariness Herb Gynostemmae Pentaphylli is grace five
Leaf honey or grace seven leaf are sweet.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 2,
It is characterized in that, described in disinfect and include that by described outer implant liquid detergent or concentration be 5%
Detergent rinsing 3～8min after, with flowing water rinse 0.5～1h, then use 75% soak with ethanol
25～35s, sterile deionized water rinses 4～6 times, and being subsequently placed into concentration is 0.1%HgCl2Solution
Middle vibration sterilization 7～10min or with hypochlorite disinfectant 22～27min, then with aseptic go from
Sub-water rinses 5～6 times.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 1,
It is characterized in that, described tissue culture, temperature be 24～26 DEG C, light application time be 11～13h/d,
Intensity of illumination is to carry out under conditions of 1500～2000lx.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 4,
It is characterized in that, described induction of callus includes described outer implant is inoculated into wound healing group
Knitting in inducing culture, described outer implant is formed callus, described callus induction is trained
Foster base be in MS culture medium add 1～2mg/L 6-BA, 0.2～0.4mg/L NAA,
20～40g/L sucrose and 6～8g/L agar prepare.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 5,
It is characterized in that, described differentiation is cultivated and is included described callus is transferred to differentiation training
Supporting in base, and differentiate Multiple Buds, described division culture medium is to add in MS culture medium
0.1～0.3mg/L NAA, 0.1～0.3mg/L 2,4-D, 0.5～1.5mg/L 6-BA, 20～40
G/L sucrose and 6～8g/L agar.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 6,
It is characterized in that, described root culture includes being transferred in root media described Multiple Buds,
Growing root, form described tissue cultured seedling, described root media is in 1/2MS culture medium
Add 1～2mg/L NAA, 0.2～0..4mg/L PP333With 90～110mg/L root-inducing powders.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 1,
It is characterized in that, when setting up described expanding propagation garden and garden, described land for growing field crops kind, respectively by described strain
It is rhizome and the described expanding propagation garden growth strand of 1 year of the Herb Gynostemmae Pentaphylli strain plant that garden grows 1 year
The rhizome of blue strain plant dilutes the agricultural streptomycin infiltration 0.5～1min of 2000 times and pulls out,
It is 1 section by 3-4 stipes, is truncated into kind of a root, plant by seeding row spacing 15cm × 15cm kind, every cave
1, set up described expanding propagation garden and garden, described land for growing field crops kind respectively.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 1,
It is characterized in that, in described Cultivate administration, in by garden, described land for growing field crops kind, grow the strand of 1 year
When the rhizome of the blue plant of stock plants land for growing field crops, first applying fertilizer land for growing field crops, described fertilising includes often
Mu uses well-rotted farmyard manure 4500～5500kg, complex micro organism fungicide fertile 1～3kg and Se content
The Se-enriched bio-organic fertilizer 70～90kg of 300mg/kg.
The implantation methods of organic selenium-rich sugariness Herb Gynostemmae Pentaphylli the most according to claim 9,
It is characterized in that, use complex micro organism fungicide in peak of growing season fertile, use in the stem and leaf collecting period
Organic selenium-rich liquid fertilizer, described organic selenium-rich liquid fertilizer is by bacillus subtilis is inoculated into test tube
Rejuvenation culture medium is cultivated and forms test tube kind, then described test tube kind is inoculated in fermentation tank and carries out sending out
Ferment prepares；Wherein, the composition of described test tube rejuvenation culture medium includes: 6g/L glucose, 5g/L sugarcane
Sugar, 5g/L Carnis Bovis seu Bubali cream, 5g/L peptone, 4g/L sodium chloride, 18g/L agar；Described fermentation
Include in tank: 6g/L glucose, 5g/L sucrose, 5g/L Carnis Bovis seu Bubali cream, 10g/L peptone, 4g/L
Sodium chloride, biofermentation defoamer 1g/L.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106332779A (en) * | 2016-08-29 | 2017-01-18 | 李军 | Tissue culture and fast propagation method for gynostemma pentaphylla |
| CN106613975A (en) * | 2016-12-15 | 2017-05-10 | 中国热带农业科学院热带作物品种资源研究所 | Mallotus oblongifolius callus culture method and culture medium thereof |
| CN108575750A (en) * | 2018-04-24 | 2018-09-28 | 镇江市德尔生物制品研究所有限公司 | Gynostemma pentaphylla tissue-culturing rapid propagation culture medium and apply its gynostemma pentaphylla tissue-culturing rapid propagation technique |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106332779A (en) * | 2016-08-29 | 2017-01-18 | 李军 | Tissue culture and fast propagation method for gynostemma pentaphylla |
| CN106613975A (en) * | 2016-12-15 | 2017-05-10 | 中国热带农业科学院热带作物品种资源研究所 | Mallotus oblongifolius callus culture method and culture medium thereof |
| CN108575750A (en) * | 2018-04-24 | 2018-09-28 | 镇江市德尔生物制品研究所有限公司 | Gynostemma pentaphylla tissue-culturing rapid propagation culture medium and apply its gynostemma pentaphylla tissue-culturing rapid propagation technique |
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| Publication number | Publication date |
|---|---|
| CN105875412B (en) | 2018-04-20 |
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