CN106258080B - A method of utilizing biotechnology vernalization Chinese yew seed - Google Patents
A method of utilizing biotechnology vernalization Chinese yew seed Download PDFInfo
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- CN106258080B CN106258080B CN201610704099.2A CN201610704099A CN106258080B CN 106258080 B CN106258080 B CN 106258080B CN 201610704099 A CN201610704099 A CN 201610704099A CN 106258080 B CN106258080 B CN 106258080B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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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
- C05F9/00—Fertilisers from household or town refuse
- C05F9/04—Biological compost
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
Abstract
The method that the present invention relates to the use of biotechnology vernalization Chinese yew seed, it is low effectively to solve Chinese yew germination emergence rate, the low problem of survival rate, and method is, by mature Chinese yew seed, to be cleaned up with clear water, is placed in desinfection chamber；60~65% biogas slurry aqueous solution of mass concentration is added according to 1 ︰ 1 of weight ratio in the seed of wash clean, the active biochemical bacteria and 0.5~1% brown sugar water of seed weight 1~2% is added, stirs evenly, 25~30 DEG C of static soaks ferment 7 days；The Chinese yew rudiment kind grain impregnated in fermentation vat is pulled out, gibberellin is added in remaining bacterium solution, as seedling stage slow release fertilizer；Seedling stage slow release fertilizer is diluted with water 5 times, the kind grain of rudiment nursery is irrigated, shades after emergence, the light transmittance 15%~20% of cool canopy, Routine Management is transplanted after 1 year.The method of the present invention is simple, easily operated, and expense is low, and effect is good, and percentage of seedgermination is high, high survival rate, ecological, environmental protective.
Description
Technical field
The present invention relates to cultivating seeds, especially a kind of method using biotechnology vernalization Chinese yew seed.
Background technique
Chinese yew is left rare or endangered species after quaternary glacier, and distribution is narrow, slow growth, existing
Quantity is reduced year by year, is the wild plant of special-protection-by-the-State.The root of Chinese yew, leaf, contains the Japanese yew that can be purified in skin at stem
Alcohol is to integrate medicinal, material use, ornamental rare tree species, has the title of " plant gold ", have very high economic development valence
Value.
Taxol has efficient anticancer activity, oophoroma and breast cancer is primarily adapted for use in, to lung cancer, colorectal cancer, melanin
Tumor, head-neck carcinoma, lymthoma, brain tumor also have certain curative effect.Structure is complicated for taxane molecule, is still difficult to realize chemical conjunction at present
At etc. factors, rely primarily on and extracted from Chinese yew histoorgan, but content of taxol is low in Chinese yew body, to Chinese yew plant
Demand is very big, but natural Chinese yew distribution is scattered, so that taxol supply problem becomes the principal element for limiting its application, because
This Efforts To Develop Chinese yew artificial cultivation becomes the task of top priority.
Chinese yew genus plants are typical feminine trees, and the variation to weather especially wants temperature, illumination and humidity
Ask relatively tight.It is good in slant acidity, draining, it moistens, well-grown on loose, fertile soil.It is educated by Taxus chinensis var. mairei Seeds
Seedling test discovery, Chinese yew produce seeds quantity it is few, kind skin depth, dormant period be up to 1 year or more, suitable autumn adopt after broadcast, sow in spring need to use
The seed by low-temperature stratification vernalization or next year buried, germinates in 2 to three years successively after otherwise broadcasting.Nursery stock primary growth is slow
Slowly, only 5~6 centimetres of annual seedling height, root is elongated, and fibrous root is less, need to stay bed 2 to three years, and can be out of the garden transplanting.Due to this
Kind shade tolerance is extremely strong, and slow growth, and seedling is advisable after occurring with appropriate shade.In short, with Chinese yew seminal propagation, emergence rate
Low, seed difficulty is adopted, and Seedling survival rate is not high, leads to seedling cost height.
Summary of the invention
For above situation, for the defect for solving the prior art, the purpose of the present invention is just to provide a kind of biological skill of utilization
The method of art vernalization Chinese yew seed, it is low effectively to solve Chinese yew germination emergence rate, the low problem of survival rate.
The technical solution that the present invention solves is, comprising the following steps:
1. seed treatment: mature Chinese yew seed being cleaned up with clear water, is placed in desinfection chamber；
2. impregnating: by the seed of wash clean according to 1 ︰ 1 of weight ratio addition 60~65% biogas slurry aqueous solution of mass concentration, then plus
The active biochemical bacteria and 0.5~1% brown sugar water for entering seed weight 1~2%, stir evenly, and 25~30 DEG C of static soaks ferment 7 days；
3. the Chinese yew rudiment kind grain impregnated in fermentation vat is pulled out, gibberellin is added in remaining bacterium solution, it is slow as seedling stage
Release fertilizer, the weight content in slow release fertilizer is 5% to gibberellin in seedling stage；
4. carrying out seeding and seedling raising to Chinese yew rudiment kind grain in nursery: seedling stage slow release fertilizer being diluted with water 5 times, is used for
The Chinese yew kind grain for irrigating rudiment nursery, will shade after emergence, and seedling is afraid of that day burns, then the light transmittance 15%~20% of cool canopy is pressed
Routine is managed, and is transplanted after 1 year.
The active biochemical bacteria is the multipurpose compound fungus of patent No. ZL200710054677.3 method preparation, specifically
It is: parent species is made with thermophilic fiber bacterium, saccharomyces cerevisiae and streptomyces rimosus first, three kinds of female strains expand and grow culture, step
It is: conventional sterilant is carried out to thermophilic fiber bacterium culture medium, then the merging of thermophilic fiber starter kind is grown equipped with the training of thermophilic fiber bacterium culture medium
In test tube, strain additional amount is 1:100 to culture medium weight ratio by strain, and thermophilic every 1000 g of fiber bacterium culture medium is by beef extract
4~15g, 15~20g of peptone, 1~3g of soluble starch, 6~8g of salt, 18~20g of agar and surplus are water composition, thermophilic fibre
After tieing up starter kind implantation culture medium, trains and grow 84 hours under being 7.3 in 26 DEG C of constant temperature, pH value, be multiplied into test tube species, then be placed in training and grow
Anaerobic fermentation is carried out in tank, at 18~30 DEG C anaerobic fermentation 200 hours it is stand-by；Conventional go out is carried out to brewer's yeast bacterium culture medium
Bacterium, the saccharomyces cerevisiae after sterilizing by weight is implanted into saccharomyces cerevisiae than the ratio that brewer's yeast bacterium culture medium is 1:100,26
Shaking flask culture 84 hours under DEG C constant temperature, at saccharomyces cerevisiae test tube species, described brewer's yeast bacterium culture medium is by grape for numerous expansion
25~35g of sugar, 15~25g of yeast powder add fermented bean drink be adjusted to 1000 ml composition, expand it is numerous at saccharomyces cerevisiae test tube species be placed in training again
Interior anaerobic fermentation 200 hours at 18~30 DEG C of tank are grown, the saccharomyces cerevisiae for expanding and growing is become, it is spare；To streptomyces rimosus culture
Base carries out 120 DEG C of high-temperature heat sterilizations, then the merging of streptomyces rimosus parent species is equipped with sterilized streptomyces rimosus culture medium
Training grow in tank, streptomyces rimosus parent species additional amount be culture medium weight 1/100, i.e., weight ratio be 1:100, pH value 7.3,
Constant temperature is to train to grow 84 hours at 26 DEG C, is multiplied into test tube species, described streptomyces rimosus culture medium is the culture medium of every 1000ml
It is by 15~25g of soluble starch, 0.5~1.5g of potassium nitrate, 0.4~0.8g of salt, K2HPO4.3H20.4~0.8g of O,
MgSO4.7H20.4~0.8g of O, FeSO4.7H20.005~0.02g of O, 15~25g of agar and surplus are made of water, and pH value is
7.2~7.4, when preparation, soluble starch is first tuned into paste with cold water, then pour into the water boiled, is heated on fire, while stirring
It mixes side and other ingredients is added, after being sufficiently uniformly dissolved, supply moisture to 1000 ml, the test tube species streptomyces rimosus being multiplied into is again
Merging training is grown in tank, 18~30 DEG C at a temperature of, carry out anaerobic fermentation 200 hours, training, which is grown into, expands the streptomyces rimosus grown, standby
With；Three kinds are expanded thermophilic fiber bacterium, saccharomyces cerevisiae and the streptomyces rimosus grown after anaerobic fermentation and are existed with weight 1:1:1 ratio mixture
Form Mixed Microbes together, then be placed in training and grow in tank, be added wheat bran or rice bran expand it is numerous, carried out at 15~3 DEG C solid expand it is numerous
240~300 hours, the weight ratio of wheat bran or rice bran and Mixed Microbes was 100:1, solid expand it is numerous after place into training and grow tank, at 25 DEG C
It ferments 200 hours under constant temperature, at bacterium powder, water content 2.5~4%, finished product packing after drying and dehydrating.
The method of the present invention is simple, easily operated, and expense is low, and effect is good, and percentage of seedgermination is high, high survival rate, and ecology
Environmental protection is the technical innovation of planting yew.
Specific embodiment
Specific embodiments of the present invention will be described in further detail with reference to embodiments.
Embodiment 1
The present invention in specific implementation, September is put in basin to the mature Chinese yew seed of acquisition in October and is cleaned with clear water
Completely, it is placed in desinfection chamber, 60% biogas slurry aqueous solution of mass concentration is added according to 1 ︰ 1 of weight ratio in the seed of wash clean, adds kind
The active biochemical bacteria and 0.5% brown sugar water of sub- weight 1%, stir evenly, and 30 DEG C of static soaks ferment 7 days.
Embodiment 2
The present invention in specific implementation, September is put in basin to the mature Chinese yew seed of acquisition in October and is cleaned with clear water
Completely, it is placed in desinfection chamber, 65% biogas slurry aqueous solution of mass concentration is added according to 1 ︰ 1 of weight ratio in the seed of wash clean, adds kind
The active biochemical bacteria and 1% brown sugar water of sub- weight 2%, stir evenly, and 25 DEG C of static soaks ferment 7 days.
By described in Examples 1 to 2, after Chinese yew Seeds preprocess, the seeding technique of Summary is as follows, it may be assumed that
1) the annual September Chinese yew seed mature to acquisition in October, be put in after being cleaned up in basin with clear water be transported into it is sterile
Room；
2) seed is put into rustless steel container, and 60~65% biogas slurry aqueous solution of mass concentration is added according to 1 ︰ 1 of weight ratio, then
The active biochemical bacteria and 0.5~1% brown sugar water of seed weight 1~2% is added, stirs evenly, 25~30 DEG C of static soak fermentations 7
It；
3) the Chinese yew rudiment kind grain impregnated in fermentation vat is pulled out, gibberellin is added in remaining bacterium solution, it is slow as seedling stage
Release fertilizer, the weight content in slow release fertilizer is 5% to gibberellin in seedling stage；
4) seeding and seedling raising is carried out to Chinese yew rudiment kind grain in nursery, the seedling stage slow release fertilizer of previous stage configuration is diluted
After 5 times, for irrigating the Chinese yew kind grain of above-mentioned nursery, to shade after emergence, seedling is afraid of that day burns, the light transmittance 15% of cool canopy~
20%, then by traditional water and fertilizer management, transplanted after 1 year.
Chinese yew seed is handled with the present invention, sowing experiment shows: the Chinese yew seed without the process is after planting
Germination percentage be only 50%；And the germination percentage of Chinese yew seed after planting through the process can reach 96%, survival rate is up to 99%
More than.
Innovation of the invention is: for the first time using discarded biogas slurry, active bacteria technology to Chinese yew presprouting of seeds nursery,
The germination percentage and survival rate for improving seed seedling-raising, are the technical innovations of planting yew.
Compared with prior art, the present invention having the advantage that
1. germination percentage and high survival rate
Pregermination and seedling breeding is carried out to Chinese yew seed using the present invention, the effective component that active biochemical bacteria generates can be quick
Seed dormancy is released, and biogas slurry is rich in multiple nutritional components, sprouts for the seed later period and provides nutrition deposit, substantially increases and educate
The germination percentage and survival rate of seedling, while active biochemical bacteria and biogas slurry can effectively kill the various pathogens hidden in seed, subtract
The light generation of later period pest and disease damage, increases nursery success rate.The fermented carrying of the pericarp and pulp of outer covering of the seed is a certain amount of
Active biochemical bacteria also can promote rudiment, resists pest and disease damage and infects.
2. it is at low cost, it is easy to operate
The present invention implements kind of a required raw material, and cost is relatively low, and compared with general chemistry pesticide, cost is only the 70% of its cost.
Moreover, the technology of preparing is easy to operate, it is easy to grasp, input and output are higher.
3. ecological friendly
The present invention implements kind of a required raw material, and main component is remaining biogas slurry after biogas fermentation, from a wealth of sources, nonhazardous, nothing
Sharp aroma is conducive to recycling for resource, alleviates the environmental pressure of biogas slurry discharge；Reduce the use of chemicals
Amount, more ecological, environmental protective.
In short, the present invention is to carry out Pregermination and seedling breeding to Chinese yew seed using biogas slurry and active biochemical bacteria, substantially increase
Chinese yew seeding ratio and survival rate are low, reduce seedling cost, and efficiently solve scale raising excrement object biogas slurry
Pollution problem, it was found that the new way that an agricultural wastes recycle meets the requirement of agricultural sustainable development, ecological, environmental protective
Characteristic is preferable, and popularization and application foreground is wide.Operation of the present invention method is simple, and product practice effect is good, no sharp aroma, environment friend
It is good, there is important application value.
Claims (3)
1. a kind of method using biotechnology vernalization Chinese yew seed, which comprises the following steps:
(1) seed treatment: mature Chinese yew seed is cleaned up with clear water, is placed in desinfection chamber；
(2) it impregnates: 60~65% biogas slurry aqueous solution of mass concentration is added according to 1 ︰ 1 of weight ratio in the seed of wash clean, adds kind
The active biochemical bacteria and 0.5~1% brown sugar water of sub- weight 1~2%, stir evenly, and 25~30 DEG C of static soaks ferment 7 days；
The active biochemical bacteria is multipurpose compound fungus, makees parent species with thermophilic fiber bacterium, saccharomyces cerevisiae and streptomyces rimosus,
Three kinds of female strains expand and grows culture and is made, step is: conventional sterilant carried out to thermophilic fiber bacterium culture medium, then by thermophilic fiber bacterium
Parent species merging is grown in test tube equipped with the training of thermophilic fiber bacterium culture medium, and strain additional amount is 1:100 to culture medium weight ratio by strain,
Thermophilic every 1000 g of fiber bacterium culture medium be by 4~15g of beef extract, 15~20g of peptone, 1~3g of soluble starch, salt 6~
8g, 18~20g of agar and surplus are water composition, after thermophilic fiber starter kind implantation culture medium, in the case where 26 DEG C of constant temperature, pH value are 7.3
Training is grown 84 hours, and test tube species are multiplied into, then is placed in training and is grown progress anaerobic fermentation in tank, and anaerobic fermentation 200 is small at 18~30 DEG C
When it is stand-by；Conventional sterilant is carried out to brewer's yeast bacterium culture medium, the saccharomyces cerevisiae after sterilizing by weight is trained than saccharomyces cerevisiae
It supports the ratio that base is 1:100 and is implanted into saccharomyces cerevisiae, shaking flask culture 84 hours under 26 DEG C of constant temperature, numerous expansion is tried at saccharomyces cerevisiae
Pipe kind, described brewer's yeast bacterium culture medium are to add fermented bean drink to be adjusted to 1000 ml groups by 25~35g of glucose, 15~25g of yeast powder
At, expand it is numerous at saccharomyces cerevisiae test tube species be placed in training again and grow in tank anaerobic fermentation 200 hours at 18~30 DEG C, become expansion and grow
Saccharomyces cerevisiae, it is spare；120 DEG C of high-temperature heat sterilizations are carried out to streptomyces rimosus culture medium, then by streptomyces rimosus parent species
Merging is grown in tank equipped with the training of sterilized streptomyces rimosus culture medium, and streptomyces rimosus parent species additional amount is culture medium weight
1/100, i.e. weight ratio is 1:100, and pH value 7.3, constant temperature is to train and grow 84 hours at 26 DEG C, is multiplied into test tube species, described
Streptomyces rimosus culture medium is that the culture medium of every 1000ml is by 15~25g of soluble starch, 0.5~1.5g of potassium nitrate, salt
0.4~0.8g, K2HPO4.3H20.4~0.8g of O, MgSO4.7H20.4~0.8g of O, FeSO4.7H20.005~0.02g of O, fine jade
15~25g of rouge and surplus are made of water, and pH value is 7.2~7.4, when preparation, and soluble starch is first tuned into paste with cold water, then
It pours into the water boiled, is heated on fire, other ingredients are added while stirring, after being sufficiently uniformly dissolved, supply moisture to 1000
Ml, the test tube species streptomyces rimosus being multiplied into are placed in training again and grow in tank, 18~30 DEG C at a temperature of, carry out anaerobic fermentation 200
Hour, the streptomyces rimosus for expanding and growing is grown into training, spare；Three kinds expand after anaerobic fermentation the thermophilic fiber bacterium grown, saccharomyces cerevisiae and
Streptomyces rimosus forms Mixed Microbes with weight 1:1:1 ratio mixture together, then is placed in training and grows in tank, be added wheat bran or rice bran into
Row expansion is numerous, and the weight ratio that numerous 240~300 hours of solid expansion, wheat bran or rice bran and Mixed Microbes are carried out at 15~3 DEG C is 100:1,
Solid expand it is numerous after place into training and grow tank, ferment 200 hours under 25 DEG C of constant temperature, at bacterium powder after drying and dehydrating, water content 2.5~
4%, finished product packing；
(3) the Chinese yew rudiment kind grain impregnated in fermentation vat is pulled out, gibberellin is added in remaining bacterium solution, as seedling stage slow-release fertilizer
Material, the weight content in slow release fertilizer is 5% to gibberellin in seedling stage；
(4) seeding and seedling raising is carried out to Chinese yew rudiment kind grain in nursery: seedling stage slow release fertilizer is diluted with water 5 times, for irrigating
The Chinese yew kind grain of rudiment nursery, will shade after emergence, and seedling is afraid of that day burns, the light transmittance 15%~20% of cool canopy, then routinely
It is managed, is transplanted after 1 year.
2. the method according to claim 1 using biotechnology vernalization Chinese yew seed, which is characterized in that described is red
The immersion of beans China fir seed is to be put in basin September to the mature Chinese yew seed of acquisition in October and cleaned up with clear water, is placed in sterile
The seed of wash clean is added 60% biogas slurry aqueous solution of mass concentration according to 1 ︰ 1 of weight ratio, adds the work of seed weight 1% by room
Property biochemical bacteria and 0.5% brown sugar water, stir evenly, 30 DEG C of static soaks ferment 7 days.
3. the method according to claim 1 using biotechnology vernalization Chinese yew seed, which is characterized in that described is red
The immersion of beans China fir seed is to be put in basin September to the mature Chinese yew seed of acquisition in October and cleaned up with clear water, is placed in sterile
The seed of wash clean is added 65% biogas slurry aqueous solution of mass concentration according to 1 ︰ 1 of weight ratio, adds the work of seed weight 2% by room
Property biochemical bacteria and 1% brown sugar water, stir evenly, 25 DEG C of static soaks ferment 7 days.
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| CN107006167A (en) * | 2017-03-17 | 2017-08-04 | 界首市兴邦家庭农场 | A kind of method for culturing seedlings of soya seeds bud rate occurred frequently |
| CN109121561A (en) * | 2018-08-13 | 2019-01-04 | 陇南远大生物科技有限公司 | Using the method for biofermentation processing Glycyrrhiza Seeds |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103283445A (en) * | 2013-05-17 | 2013-09-11 | 旌德县岭南园林绿化有限公司 | Method for comprehensively applying biogas and biogas manure to seedling growth of flower and tree seeds |
| CN104335794A (en) * | 2013-07-28 | 2015-02-11 | 赵百华 | Pot culture technique of tcaus mairei for both medicinal and viewing function |
| CN105123018A (en) * | 2015-07-30 | 2015-12-09 | 黎怀玲 | Chinese yew seed pregermination method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283445A (en) * | 2013-05-17 | 2013-09-11 | 旌德县岭南园林绿化有限公司 | Method for comprehensively applying biogas and biogas manure to seedling growth of flower and tree seeds |
| CN104335794A (en) * | 2013-07-28 | 2015-02-11 | 赵百华 | Pot culture technique of tcaus mairei for both medicinal and viewing function |
| CN105123018A (en) * | 2015-07-30 | 2015-12-09 | 黎怀玲 | Chinese yew seed pregermination method |
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