CN104161047B - The preparation method of transpiration inhibitor based on spirulina induction pore immunity Guan Bi - Google Patents
The preparation method of transpiration inhibitor based on spirulina induction pore immunity Guan Bi Download PDFInfo
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- CN104161047B CN104161047B CN201410326611.5A CN201410326611A CN104161047B CN 104161047 B CN104161047 B CN 104161047B CN 201410326611 A CN201410326611 A CN 201410326611A CN 104161047 B CN104161047 B CN 104161047B
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- spirulina
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
Abstract
The preparation method of the open transpiration inhibitor based on spirulina induction pore immunity Guan Bi of the present invention.First being inoculated into by fresh water spirulina in the liquid spirulina culture medium after sterilizing, be placed in illumination box cultivation, be subsequently placed in centrifuge centrifugal, dilution obtains pure spirulina solution；Be placed in 121 DEG C, 0.12～0.15 Mpa pressure high-pressure sterilizing pot in inactivation make spirulina agent A；By CaCl2·2H2O and sucrose are dissolved in distilled water, stir, and obtain adjuvant B；Spirulina agent A is joined in adjuvant B, stirring and evenly mixing.The present invention has the advantages such as recyclability, low cost (spirulina is the microorganism that photoautotrophy produces), environment friendly and high efficiency, belong to a new generation's microorganism induction type transpiration suppression water-saving agent, there is powerful application prospect and potential market competitiveness.
Description
Technical field
The invention belongs to biological field, be specifically related to a kind of based on spirulina induction pore immunity Guan Bi
The preparation method of transpiration inhibitor.
Background technology
Along with Global climate change and population increase, the shortage of freshwater resources has become the weight that the whole world faces
Big challenge.Agricultural water proportion is always bigger.Therefore, exploitation novel plant is rising
Suppression water-saving agent, effectively controls the aperture of pore, reduces the excessive rising of blade, improves moisture profit
By efficiency, it is to solve water resources in china crisis and the effective way of Developing Water-saving Agriculture.According to it not
Same model of action and feature, plant transpiration inhibitor in the market can be divided into 4 classes: 1. becomes
Membranous type antiperspirant (Film-forming antitranspirant), common film-forming type medicament has
Wilt-Pruf, Vapor Guard, Mobileaf, Folicote, Plant guard etc., greatly
It is all the patented product of expensive foreign, is difficult to natural degradation, there is persistently pollution.2. reflection-type resists
Rising agent (Reflecting antitranspirant), conventional has: Kaolin (Kaoline)
With kaolinite (Kaolinite), the usage amount of this type of preparation is big, weak effect and have directly fall
The side effect etc. of low photosynthetic rate, the most less actual application.3. metabolic pattern antiperspirant
(Metabolic antitranspirant), conventional have PMA (phenylmercuric acetate), ABA,
NaHSO3、CaC12, CCC, 2,4-D, G-30027, alachlor, triazolone, fulvic acid (FA),
Wherein mostly there is in addition to fulvic acid serious environmental toxicity (such as phenylmercuric acetate) and physiological-toxicity, danger
Evil environment and human health, the growth regulator such as ABA not only has environmental toxicity but also price simultaneously
Costliness, is therefore difficult to promote the use of.4. biological response type antiperspirant (biological
Induction antitranspirant), the one of its Shi Wanggen pavilion et al. invention is based on pore sense
Answer the biotype plant transpiration resistant agent of microorganism, but owing to microorganism may produce some, plant is had
The material of side effect, that also needs to further improve just to be actually applied popularization.Therefore at water
Under conditions of crisis of resource is day by day serious and drought takes place frequently, in the urgent need to researching and developing further newly
The anti-rising preparation of biological response type of type high effect nontoxic and water-saving technology.
Summary of the invention
It is an object of the invention to for the deficiencies in the prior art, it is provided that a kind of based on spirulina induction
The preparation method of the transpiration inhibitor of pore immunity Guan Bi.
The inventive method comprises the following steps:
Step (1). prepare spirulina agent A:
First fresh water spirulina (Spirulina platensis) is inoculated into the liquid spiral shell after sterilizing
In rotation algae culture medium, being placed in illumination box cultivation, cultivation temperature controls at 25～30 DEG C, training
Support 8～10 periodicity of illuminations, a periodicity of illumination is 24 hours, initially with light intensity be 2500～
The light illumination 12h of 3500lx, then it is placed in 12h in dark, the spirulina liquid after being cultivated；
Then the spirulina liquid after cultivating is placed in a centrifuge centrifugal, and rotating speed is 2000～2500 revs/min,
Taking washing of precipitate after Li Xin and be dried, being then diluted with distilled water, obtaining concentration is 10～15
The pure spirulina solution of g/L；The pure spirulina liquid prepared is placed in 121 DEG C, 0.12～0.15
The high-pressure sterilizing pot of Mpa pressure inactivates 70～90 minutes, after cooling, frozen in-20 DEG C,
Make spirulina agent A；
Described fresh water spirulina (Spirulina platensis) is bought in Chinese Academy of Sciences's allusion quotation
Type culture collection committee's algae kind storehouse, deposit number is FACHB-439；
Described liquid spirulina culture medium is solution I, solution II, vitamin B12Mixed liquor；
Wherein solution I is 1:1 with the volume ratio of solution II, vitamin B12Concentration be 5 × 10-6g/L；
Solution I is by sodium bicarbonate 13.61g/500ml, sodium carbonate 4.03g/500ml, phosphoric acid hydrogen two
Potassium 0.5g/500ml preparation obtains；Solution II is by sodium nitrate 2.50g/500ml, sodium chloride 1.00
G/500ml, potassium sulfate 1.00g/500ml, magnesium sulfate 0.20g/500ml, calcium chloride 0.04
G/500ml, ferrous sulfate 0.01g/500ml, ethylenediaminetetraacetic acid 0.08g/500ml join
Prepare；
Step (2). prepare adjuvant B:
By CaCl2·2H2O and sucrose are dissolved in distilled water, stir, and obtain adjuvant B；Auxiliary
CaCl in agent B2·2H2The concentration of O is 2g/L, and the concentration of sucrose is 0.4g/L；
Step (3). prepare transpiration inhibitor:
Step (1) spirulina agent A is joined in step (2) adjuvant B, stirring and evenly mixing；Its
The volume ratio of middle step (1) spirulina agent A and step (2) adjuvant B is 0.03～0.06:1.
Present invention utilizes the principle of microorganism induction immunity Guan Bi.Specifically, plant stomata sense
Answer the microorganism can closedown actively so that it is phenomenon pathogenic in plant can not being entered by pore
It is referred to as pore immunity.It is micro-that its mechanism is that the specific receptor on plant stomata guard cell is capable of identify that
Biological or come from the associated molecular pattern of microorganism, cause the signal level that a series of mediation pore is closed
Connection reaction, including kinase whose activation, the generation of second message,second messenger, the vibration of calcium ion and raising, egg
White modification, the restructuring of cytoskeleton, the rising of cytoplasmic matrix PH, ion and the tune of aquaporin
Joint.In nature, plant stomata plays most important in the transpiration and photosynthesis controlling plant
Effect, along with stomatal aperture reduces in certain threshold value, transpiration rate generally subtracts than photosynthetic rate
Few is fast, so the suitable aperture reducing pore, can improve the instantaneous water utilization effect of plant
Rate.This senses the rising suppression water-saving agent offer of microorganism and immunity Guan Bi for the pore that exploitation is novel
Theoretical basis.Found by research, spray certain density spirulina permissible to the blade of plant
Cause plant stomata aperture (stomatal aperture/pore length) and the significantly decline of stomatal conductance, and then
Increasing stomatal resistance, the transpiration decreasing moisture is scattered and disappeared, and improves plant over a period to come
WUEL.
The invention have the advantages that:
1. suppression transpiration water saving preparation based on pore sensing microorganism immunity Guan Bi mechanism, ratio is all kinds of
Existing transpiration inhibitor has more high-efficiency water-saving, safety and environmental protection, low cost and adaptation and gives birth on a large scale
The advantage produced.
2. spirulina is photoautotrophy microalgae, does not only have carbon emission, also can form new carbon and converge,
It has the highest nutritive value and pharmacodynamics effect simultaneously.
3. water-saving result is notable, applied widely, is that the preferable plant in arid and semi-arid lands is steamed
Rise suppression water-saving agent.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further analyzed.
Embodiment 1.
Step (1). the preparation of spirulina agent A, method is:
First fresh water spirulina (Spirulina platensis) is inoculated into the liquid after sterilizing
In Spirulina culture medium, being placed in illumination box cultivation, light intensity is set to 3500lx, illumination
Cycle is that 12h illumination/12h is dark, and cultivation temperature controls at 25 DEG C, cultivates 8 days, is trained
Spirulina liquid after Yanging；
Then the spirulina liquid after cultivating is placed in a centrifuge centrifugal, rotating speed 2000 revs/min, from
After the heart, washing and be dried, and then compound concentration is the spirulina solution that 15g/L is pure.Will preparation
Good pure spirulina liquid is placed on 121 DEG C, inactivates 70 points in the high-pressure sterilizing pot of 0.15 MPa
Clock, after cooling, frozen in-20 DEG C, make spirulina agent A；
Step (2). preparing adjuvant B, method is:
By 2g CaCl2·2H2O and 0.4g sucrose is dissolved in 1L distilled water, stirs,
Obtain adjuvant B；
Step (3). preparing transpiration inhibitor, method is: by 0.03L step (1) spirulina agent A
Join in 1L step (2) adjuvant B, stirring and evenly mixing.
Embodiment 2
Step (1). the preparation of spirulina agent A, method is:
First fresh water spirulina (Spirulina platensis) is inoculated into the liquid after sterilizing
In Spirulina culture medium, being placed in illumination box cultivation, light intensity is set to 3000lx, illumination
Cycle is that 12h illumination/12h is dark, and cultivation temperature controls at 28 DEG C, cultivates 9 days；
Then cultured spirulina liquid is placed in a centrifuge centrifugal, rotating speed 2200 revs/min, from
After the heart, washing and be dried, and then compound concentration is the spirulina solution that 13g/L is pure.Will preparation
Good pure spirulina liquid is placed on 121 DEG C, inactivates 80 points in the high-pressure sterilizing pot of 0.13 MPa
Clock, after cooling, frozen in-20 DEG C, make spirulina agent A；
Step (2). preparing adjuvant B, method is:
By 2g CaCl2·2H2O and 0.4g sucrose is dissolved in 1L distilled water, stirs,
Obtain adjuvant B；
Step (3). preparing transpiration inhibitor, method is: by 0.04L step (1) spirulina agent A
Join in 1L step (2) adjuvant B, stirring and evenly mixing.
Embodiment 3
Step (1). the preparation of spirulina agent A, method is:
First fresh water spirulina (Spirulina platensis) is inoculated into the liquid after sterilizing
In Spirulina culture medium, being placed in illumination box cultivation, light intensity is set to 2500lx, illumination
Cycle is that 12h illumination/12h is dark, and cultivation temperature controls at 30 DEG C, cultivates 10 days；
Then cultured spirulina liquid is placed in a centrifuge centrifugal, rotating speed 2500 revs/min, from
After the heart, washing and be dried, and then compound concentration is the spirulina solution that 10g/L is pure.Will preparation
Good pure spirulina liquid is placed on 121 DEG C, inactivates 90 points in the high-pressure sterilizing pot of 0.12 MPa
Clock, after cooling, frozen in-20 DEG C, make spirulina agent A；
Step (2). preparing adjuvant B, method is:
By 2g CaCl2·2H2O and 0.4g sucrose is dissolved in 1L distilled water, stirs,
Obtain adjuvant B；
Step (3). preparing transpiration inhibitor, method is: by 0.06L step (1) spirulina agent A
Join in 1L step (2) adjuvant B, stirring and evenly mixing.
During use, the antiperspirant by sterilized water (comparison) with according to three kinds of embodiment method preparations sprays
It is applied to the blade surface of plant.First by above-mentioned 3 kinds of embodiments preparation antiperspirant, then distinguish
Spray the antiperspirant of sterilized water and the preparation of 3 kinds of embodiments to live body Semen Viciae fabae, after 24 hours,
The width of 50 pores of live body random measurement and length under microscope, simultaneously with the photosynthetic instrument of Li-6400
Measure the stomatal conductance of blade, photosynthetic rate, transpiration rate, and with photosynthetic rate and transpiration rate
Ratio calculate WUEL, result meansigma methods and standard error represent.Such as following table:
The transpiration inhibitor of three embodiment method preparations can significantly reduce Semen Viciae fabae as seen from the above table
Stomatal aperture (stomatal aperture/pore length), stomatal conductance and improve Broad Bean Leaves water utilization
Efficiency, reaches the effect of good suppression plant transpiration.
The fresh water spirulina (Spirulina platensis) that above-described embodiment uses is bought in China
American Type Culture Collection committee of academy of science algae kind storehouse, deposit number is FACHB-439；
The liquid spirulina culture medium used is solution I, solution II, vitamin B12Mixed liquor；
Wherein solution I is 1:1 with the volume ratio of solution II, vitamin B12Concentration be 5 × 10-6g/L；
Solution I is by sodium bicarbonate 13.61g/500ml, sodium carbonate 4.03g/500ml, phosphoric acid hydrogen two
Potassium 0.5g/500ml preparation obtains；Solution II is by sodium nitrate 2.50g/500ml, sodium chloride 1.00
G/500ml, potassium sulfate 1.00g/500ml, magnesium sulfate 0.20g/500ml, calcium chloride 0.04
G/500ml, ferrous sulfate 0.01g/500ml, ethylenediaminetetraacetic acid 0.08g/500ml join
Prepare.
Claims (1)
1. the preparation method of transpiration inhibitor based on spirulina induction pore immunity Guan Bi, it is characterised in that should
Method comprises the following steps:
Step (1). prepare spirulina agent A:
First fresh water spirulina Spirulina platensis is inoculated into the liquid spirulina after sterilizing cultivate
In base, being placed in illumination box cultivation, cultivation temperature controls at 25～30 DEG C, cultivates 8～10 illumination
In the cycle, a periodicity of illumination is 24 hours, initially with the light illumination 12h that light intensity is 2500～3500lx,
It is placed in 12h in dark again, the spirulina liquid after being cultivated；Then will cultivate after spirulina liquid be placed in from
Being centrifuged in scheming, rotating speed is 2000～2500 revs/min, takes washing of precipitate and be dried after being centrifuged, then with steaming
Distilled water is diluted, and obtains the pure spirulina solution that concentration is 10～15g/L；The pure spirulina that will prepare
Liquid is placed in the high-pressure sterilizing pot of 121 DEG C, 0.12～0.15Mpa pressure inactivation 70～90 minutes, treats cold
But after, frozen in-20 DEG C, make spirulina agent A；
Described fresh water spirulina Spirulina platensis buys and protects in Chinese Academy of Sciences's type culture
Hiding committee's algae kind storehouse, deposit number is FACHB-439；
Step (2). prepare adjuvant B:
By CaCl2·2H2O and sucrose are dissolved in distilled water, stir, and obtain adjuvant B；In adjuvant B
CaCl2·2H2The concentration of O is 2g/L, and the concentration of sucrose is 0.4g/L；
Step (3). prepare transpiration inhibitor:
Step (1) spirulina agent A is joined in step (2) adjuvant B, stirring and evenly mixing；Wherein step
(1) volume ratio of spirulina agent A and step (2) adjuvant B is 0.03～0.06:1；
Described liquid spirulina culture medium is solution I, solution II, vitamin B12Mixed liquor；The most molten
Liquid I is 1:1 with the volume ratio of solution II, vitamin B12Concentration be 5 × 10-6g/L；Solution I is by carbonic acid
Hydrogen sodium 13.61g/500ml, sodium carbonate 4.03g/500ml, dipotassium hydrogen phosphate 0.5g/500ml prepare
Obtain；Solution II is by sodium nitrate 2.50g/500ml, sodium chloride 1.00g/500ml, potassium sulfate 1.00g/500
Ml, magnesium sulfate 0.20g/500ml, calcium chloride 0.04g/500ml, ferrous sulfate 0.01g/500ml,
Ethylenediaminetetraacetic acid 0.08g/500ml preparation obtains.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410326611.5A CN104161047B (en) | 2014-07-10 | 2014-07-10 | The preparation method of transpiration inhibitor based on spirulina induction pore immunity Guan Bi |
| PCT/CN2014/088253 WO2016004688A1 (en) | 2014-07-10 | 2014-10-10 | Preparation method of transpiration inhibitor based on spirulina-induced immune response pore closure |
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| CN201410326611.5A CN104161047B (en) | 2014-07-10 | 2014-07-10 | The preparation method of transpiration inhibitor based on spirulina induction pore immunity Guan Bi |
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| CN104161047B true CN104161047B (en) | 2016-12-07 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106631412A (en) * | 2016-12-11 | 2017-05-10 | 孙祎 | Preparation method of non-toxic degradable transpiration inhibiter |
| CN108398429B (en) * | 2018-03-07 | 2020-08-04 | 天津农学院 | Plant stomata monitoring system and method for analyzing response of plant stomata to environment by using same |
| CN108902182B (en) * | 2018-09-28 | 2020-07-14 | 常州谙美生物科技有限公司 | Microecological preparation with function of reducing plant transpiration and preparation method thereof |
| CN109336702A (en) * | 2018-12-13 | 2019-02-15 | 河南省莱恩坪安园林植保有限公司 | A kind of plant transpiration inhibitor containing amino acid |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102626111A (en) * | 2012-03-28 | 2012-08-08 | 浙江大学 | Preparation method of immune transpiration inhibitor based on closed stomatal induction chlorella vulgaris |
| CN103387940A (en) * | 2013-07-31 | 2013-11-13 | 鄂尔多斯市新宇力藻业集团有限公司 | Culture medium and culture of Erdos plateau alkali lake spirulina platensis and culture medium preparation |
| CN103627639A (en) * | 2013-12-06 | 2014-03-12 | 张德荣 | Method utilizing spirulina cultivation liquid to cultivate Dunaliella salina |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101041812A (en) * | 2007-01-22 | 2007-09-26 | 浙江大学 | Preparation method for plant transpiration resistant agent based on escherichia coli |
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2014
- 2014-07-10 CN CN201410326611.5A patent/CN104161047B/en not_active Expired - Fee Related
- 2014-10-10 WO PCT/CN2014/088253 patent/WO2016004688A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102626111A (en) * | 2012-03-28 | 2012-08-08 | 浙江大学 | Preparation method of immune transpiration inhibitor based on closed stomatal induction chlorella vulgaris |
| CN103387940A (en) * | 2013-07-31 | 2013-11-13 | 鄂尔多斯市新宇力藻业集团有限公司 | Culture medium and culture of Erdos plateau alkali lake spirulina platensis and culture medium preparation |
| CN103627639A (en) * | 2013-12-06 | 2014-03-12 | 张德荣 | Method utilizing spirulina cultivation liquid to cultivate Dunaliella salina |
Non-Patent Citations (3)
| Title |
|---|
| 利用废水培养螺旋藻（Spirulina platensis）的实验;骆育敏等;《暨南大学学报》;19970228;第18卷(第1期);第104-109页 * |
| 可利用微藻的种类及其应用前景;张建民等;《资源开发与市场》;20051231;第21卷(第1期);第65,66,80页 * |
| 叶际微生物诱发气孔免疫的机制及其应用前景;王洋等;《植物学报》;20131231;第48卷(第6期);第658-664页 * |
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| WO2016004688A1 (en) | 2016-01-14 |
| CN104161047A (en) | 2014-11-26 |
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