CN108471713A - Plant cultivation method using metal nanoparticle and the nutrient medium applied in fact - Google Patents

Plant cultivation method using metal nanoparticle and the nutrient medium applied in fact Download PDF

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CN108471713A
CN108471713A CN201680068482.1A CN201680068482A CN108471713A CN 108471713 A CN108471713 A CN 108471713A CN 201680068482 A CN201680068482 A CN 201680068482A CN 108471713 A CN108471713 A CN 108471713A
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nutrient medium
particle
nano
zinc
copper
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CN108471713B (en
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赵辉
刘敏
陈瑜
鹿金颖
李华盛
孙乔
G·S·尼奇塔洛
A·N·日加奇
I·O·利普尼斯基
A·A·雅克米托娃
O·A·博格斯洛夫斯卡
N·N·格卢先科
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Emanuel Inst Of Biochemical Physics Of Russian Acad Of Sciences (ibcp Ras)
V L Talrose Inst For Energy Problems Of Chemical Physics Of Russian Acad Of Science (inepcp Ras)
Space Shenzhou Biology & Technology Group Co ltd
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Emanuel Inst Of Biochemical Physics Of Russian Acad Of Sciences (ibcp Ras)
V L Talrose Inst For Energy Problems Of Chemical Physics Of Russian Acad Of Science (inepcp Ras)
Space Shenzhou Biology & Technology Group Co ltd
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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
    • 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
    • A01G31/00—Soilless cultivation, e.g. hydroponics
    • 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
    • A01G22/05—Fruit crops, e.g. strawberries, tomatoes or cucumbers
    • 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
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
    • 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
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A01N59/16—Heavy metals; Compounds thereof
    • 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
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A01N59/16—Heavy metals; Compounds thereof
    • A01N59/20—Copper
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Botany (AREA)
  • Health & Medical Sciences (AREA)
  • Inorganic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Nanotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Dentistry (AREA)
  • Plant Pathology (AREA)
  • Pest Control & Pesticides (AREA)
  • Agronomy & Crop Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Medical Informatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Cultivation Of Plants (AREA)
  • Fertilizers (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

Biology in being cultivated the present invention relates to plant and field of nanometer technology can be used for producing the proportion of crop planting material of high quality, for implementing biotechnology research, so as to improve the quality of agricultural product.The present invention can be used for aerial fog cultivation method and water culture, moreover it can be used to create life support system under the conditions of long-term space flight for astronaut.The plant cultivation method proposed includes making germination on sterile letheen nutrient medium and then cultivating plant, which contains organic and inorganic constituents necessary to one group of plant growth, including：Iron, zinc and copper, wherein iron, zinc or copper or combinations thereof are the nanoparticle form of electroneutral metal.In addition, the nutrient medium may include chitosan.The present invention can improve germination and improve the physiology and morphological index of plant, such as root long degree and root activity, the chlorophyll content of blade, germination length and green portion Biomass yield.

Description

It is trained using the plant cultivation method of metal nanoparticle and the nutrition applied in fact Support base
Technical field
Biology in being cultivated the present invention relates to plant and field of nanometer technology can be used for producing the proportion of crop planting of high-quality Material, to instruct biotechnology research, so as to improve the quality of agricultural product.The present invention can be used for aerial fog cultivation method and water culture, It can be used for creating life support system under the conditions of long-term space flight for astronaut.
Background technology
Application of micron is that plant protection product and fertilizer help to improve plant and supported to adverse weather condition and disease Drag improves the yield and quality of crops.Nanotechnology preparation (for example, NanoGro, GreenLift, AgBion etc.) is Know, but the research of the method for the development field and utilization nanotechnology raising agricultural product production efficiency for novel formulation also exists Continue.Latest developments (Azamal, Husen the Khwaja Salahuddin in the field are especially summarized in survey article Siddiqi."Phytosynthesis of nanoparticles:concept,controversy and application",Nanoscale Res.Lett.,2014,V.9,№1,p.229；L.R.Khot et al." Applications of nanomaterials in agricultural production and crop protection: A review",Crop Protection,V.35,May 2012,p.64)。
Describe in application WO2013121244A1 (on 08 22nd, 2013 open) by with micro- nourishing additive agent or The nano fertilizer of metal nanoparticle (NP) form of the coating substance of their precursors.According to the invention, in extensive substance extremely A kind of few ingredient (including other elements necessary to carbon compound, phosphorus, nitrogen, boron and the nutrition of plant and development；Further include Salt, chelate and metal oxide or other compounds) it can be used as coating.In the technical scheme, metal nanoparticle (example Such as noble metal, iron, copper) be used as it is micro- fertilizer enter plant tissue and cell " delivery vehicle ", but be unable to estimate own as The independent action of nutritional ingredient.
Processing method (UA on 07 10th, 33863,2008 open) and cereal and the vegetable crop of seed has been described Cultural method (UA on December 10th, 92875,2010 is open and UA is open on December 10th, 92876,2010) includes：It uses Nano-particle blend and chemicals (its known biological action) group of biologic trace element (zinc, manganese, iron, copper, molybdenum, cobalt) The colloidal solution of conjunction.With before the case where it is similar, in these technical solutions, do not find also nano-particle as nutritional ingredient Independent role.In addition, it, which is introduced into organic products composition, is likely to result in environmental pressure, and the quality of the product is generated Adverse effect.
It is known that, conventionally, nanometer formulation be intended to stimulation plant in the open growth with protected soil.Meanwhile it is modern raw The development of object technology and utilization synthetic medium (media) (there is balanced nutrients ingredient necessary to the appropriate growth and development of plant) Planting plants and cultured tissue culture are inseparable.
With to clone plant it is quick breeding and under plant prevailing conditions form mass production breeds (hard firmly Propagated) the needs of plant generations, and with to planting material quality modern needs (need to make improvements and Test), make biotechnology be widely used with to precious species gleanings creation and maintenance it is related.
In addition, in the exploitation of spontaneous life support system (for example, being used for long-term space flight), artificial nutrition culture medium Use be very important.
Assuming that the solution of all these tasks has used advanced technology, improves plant cultivation technology and be free of disease to obtain Poison, fungi and bacterial disease, acarid and the vegetable material of nematode.
In this respect, in different technologies using nanostructure improving and cultivate planting material and be possibly used for planting The clonal propagation of object is up-and-coming route.Since culture factor and condition are important the growth of plant, nanometer is used Structure is with regard to even more important.In addition to light and temperature, the ingredient of nutrient medium is also very important factor for the culture of plant； Therefore, the culture medium variation caused by introducing nanostructure types should be able to influence the mass parameter of plant growth.
It is known that titania nanoparticles (nano-anatase) are added to Murashing- with the concentration of 10-40mg/L It can be used for increasing the germination of parsley in Skoog culture mediums (hereinafter referred to as MS culture mediums) and improve seedling quality (Dehkourdi E H.Mosavi M.Biol Trace Elem Res.2013,Nov.,155(2),p.283).It is a concentration of The TiO of 30mg/L2Nano-particle shows best effect：The germination index and survival potential for improving seed, increase root With the length of bud, green portion biomass (green mass) and other parameters are improved.However, due to still suspecting dioxy Change safety of the titanium nano-particle for living organism, can not possibly still consider to be regarded as plant growth in production estimation can Receive stimulant.It is that the mankind are potential for example, it has been shown that sucking titania powder increases the possibility of rat developing cancer Carcinogenic factor (http://www.neboleem.net/dioksid-titana.php).
According to one group of essential feature, to invent EP 2499107A1 (09 month 2012 19 disclosures) as the use stated Method prototype of the nano-particle as active factors culture plant.The invention is shown, by multilayer carbon nanotube with effective concentration 0.01-0.2pg/mL is introduced into MS culture mediums, is accelerated the germination of tomato seeds and is improved germination percentage.With control group It compares, the tomato plants being planted on the nutrient medium containing carbon nanotube have larger biomass volume, while in root There is no difference with control group in length.It is associated that this effect is improved the process of seed water suction by author with carbon nanotube auxiliary. It is known, however, that carbon nanotube passes through skin, by the way that in sucking or oral osmotic to human body, meeting image-stone cotton is equally destroyed carefully Born of the same parents.Nanotube gos deep into tissue and destroys their ability to immune system without leaving chance that canceration effect (Shvedova can be caused A.A.,Kisin E.R.,Yanamala N.,Tkach A.V.et.all."MDSC and TGFβare required for facilitation of tumor growth in the lungs of mice exposed to carbon nanotubes",Cancer Research,2015,V.75(8),pp.1-9).Therefore, it is necessary to some special researchs to be planted to control The distribution of carbon pipe, accumulation and removal in object, and apply it in the practice of plant culture.This to receive using carbon in agricultural Mitron will not be realized in a short time.
Known Nutrient medium (for example, Murashing-Skoog, Gamborg, Heller etc.) is contained necessary to development of plants The balanced nutrients compound of neccessary composition, this includes organic matter (vitamin, carbohydrate, amino acid and/or proteolysis production Object) and inorganic salts (including nitrogen, phosphorus, boron, potassium, calcium, magnesium, sulphur, iron and trace element, such as manganese, zinc, copper, molybdenum etc.).For reality The nutrient medium prototype for applying stated method is widely used Murashing-Skoog nutrient mediums (MS culture medium) (Murashing T.,Skoog F."A received medium for rapid growth and bio-assays with tobacco tissue culture".Physiol.Plant.1962.V.15,pp.473-497).Table 1 shows the prototype Agar nutrient medium known composition.
Table 1：Murashing-Skoog culture media compositions
As a part for coenzyme, metal is in the adjusting and redox for participating in plant as such as iron, copper and zinc Element is necessary in journey.MS culture mediums contain in the form of salt ion existing for these metals.
Invention content
The method that it is an object of the present invention to provide a kind of to cultivate plant on nutrient medium, the nutrient medium Nano-particle containing essential elements, the nano-particle can improve germination, and can improve form and/or the life of plant Manage parameter, it is therefore an objective to obtain improved high quality planting material.
The task is solved by the plant cultivation method proposed, and this method includes：Aseptically, containing receiving Make germination on the agar nutrient medium of rice corpuscles and then cultivate plant, wherein the nano-particle is iron nanoparticle Son or zinc nano-particle or copper nano-particle or combination thereof.
It is another object of the present invention to the nutrient medium compositions proposed for implementing method claimed.
The task is solved by the substitute of the agar nutrient medium of proposition, and the substitute contains Murashing- Ingredient contained in Skoog culture media compositions, that is, vitamin：РР、В6And В1, glycine, sucrose, meso inositol, Chelating agent Na2EDTA×2H2O, inorganic salts：NH4NO3、KNO3、CaCl2×2H2O、MgSO4×7H2O、KH2PO4、KI、H3BO3、 MnSO4×7H2O、NaMoO4×2H2O and CoCl2×6H2O, Yi Jitie, zinc and copper, wherein iron or zinc or copper or they Combination is included in the battalion in the form of Fe nanometer particles or zinc nano-particle or copper nano-particle or combination thereof respectively It supports in culture medium.
In addition, the nutrient medium can include chitosan.
In the nutrient medium, a concentration of 10.0-0.06mg/L of the nano-particle of electroneutral iron.
In the nutrient medium, a concentration of 3.0-0.016mg/L of the nano-particle of electroneutral zinc.
In the nutrient medium, a concentration of 0.04-0.00016mg/L of the nano-particle of electroneutral copper.
The solution have the advantages that stimulating germination and improving the form and/or physiological parameter of planting plants.
The essence of the present invention is actually that plant is cultivated on conventional nutrient media, in addition to improving in this way：It will The salt of necessary metal, especially ferric sulfate or zinc sulfate or copper sulphate or ferric sulfate, zinc sulfate and copper sulphate use these simultaneously The nano-particle of metal electroneutral state replaces, while other ingredients of nutrient medium remain unchanged, and in unmodified battalion It supports the plant planted on culture medium to compare, the present invention causes the seed for the plant being planted on these culture mediums more preferable can must send out Bud, and improve the form and/or physiological parameter of the plant.
The present invention is the research of the influence about metal nanoparticle to the structure and function of various biosystems with author As a result based on.(Glushchenko N.N.,BogoslovskayаO.A.,OlhovskayаI.P."Physical and chemical regularities of biological effect of high-disperse powders of Metals ", Chemical physics.2002, V.21, № 4, P.79-85；Publications on the website: http://nanobiology.narod.ru)。
It has been proved that the metal nanoparticle of electroneutral is characterized in that multi-functional and long-acting, hypotoxicity (than corresponding Ionic species metal it is 7-50 times low), and can be with biological dose (that is, 10-50 times lower than maximum tolerated dose dosage) Energetically by the organ of plant and tissue, to stimulate life process.
Further, it is known that given birth to as the absorbent of plant and microbiologic population's growth metabolism product and as plant The chitosan of long stimulating factor can mix in claimed nutrient medium.Known chitosan there is high absorption to live bacterium Property, growth-promoting effect is shown, the adaptability of plant is improved, protects the plants from infection, promotes higher crop yield (Nyanikova G.G.,Mametnabiev T.E.,Kalinkin I.P.,Gepetskaya M.V.,Komissarchik S.M.,Eldinova E.Y."Applications of chitosan"http://science.spb.ru/).It uses herein Be chitosan Taynshi (China), which is widely used in as anesthetic in Chinese medicine.Once chitosan is added to battalion It supports in culture medium, any bacteria flora, fungi or mould contamination would not occur.(as follows) in some cases, record The synergistic effect of coefficient chitosan and metal nanoparticle in nutrient medium, causes to further improve at this The germination for the plant planted under the conditions of a little and physiology and Morphologic Parameters.
Description of the drawings
It is listed below the attached drawing and its brief description for explaining present invention essence.
Figure 1A -1C show the electron microscope image and metal of Fe nanometer particles, zinc nano-particle, copper nano-particle The block diagram of particle size distribution：1A- Fe nanometer particles；1B- copper nano-particles；1C- zinc nano-particles.
Fig. 2A -2D show various concentration metal nanoparticle and chitosan (100mg/L) to Venice tomato seeds The influence of germination：2A- contains the nutrient medium of Fe nanometer particles；2B- contains the nutrient medium of zinc nano-particle；2C- contains There is the nutrient medium of copper nano-particle；2D- contains the nutrition of the combination of Fe nanometer particles, zinc nano-particle and copper nano-particle Culture medium.
Fig. 3 A-3D show various concentration metal nanoparticle and chitosan (100mg/L) to LJ-king pepper root longs The influence of degree：3A- contains the nutrient medium of Fe nanometer particles；3B- contains the nutrient medium of zinc nano-particle；3C- contains The nutrient medium of copper nano-particle；3D- contains the nutrition training of the combination of Fe nanometer particles, zinc nano-particle and copper nano-particle Support base.
Fig. 4 shows the LJ-king pepper plants planted on the nutrient medium containing zinc nano-particle and chitosan Photo：Zn-b0：Reference material；Zn-b2：The nutrient medium of zinc nano-particle containing a concentration of 0.08mg/L；Zn-b3：Contain There is the nutrient medium of the zinc nano-particle of a concentration of 0.016mg/L；Zn-b4：Zinc nano-particle containing a concentration of 0.4mg/L Nutrient medium.
Fig. 5 A-5D show various concentration metal nanoparticle and chitosan (100mg/L) to HY-2 tomato root length Influence：5A- contains the nutrient medium of Fe nanometer particles；5B- contains the nutrient medium of zinc nano-particle；5C- contains copper The nutrient medium of nano-particle；5D- contains the nutrition culture of the combination of Fe nanometer particles, zinc nano-particle and copper nano-particle Base.
Fig. 6 A-6D show various concentration metal nanoparticle and chitosan (100mg/L) to LJ-king peppers, HY- The active influence of the root of 2 tomatoes and Venice tomatoes：6A- contains the nutrient medium with Fe nanometer particles；6B- contains zinc nanometer The nutrient medium of particle；6C- contains the nutrient medium of copper nano-particle；6D- contain Fe nanometer particles, zinc nano-particle and The nutrient medium of the combination of copper nano-particle.
Fig. 7 A-7C show that the metal nanoparticle of various concentration and chitosan (100mg/L) are green to the plant leaf blade middle period The influence of cellulose content：7A- is used for the nutrient medium containing Fe nanometer particles of LJ-king peppers；7B- is for HY-2 tomatoes Nutrient medium containing zinc nano-particle；7C- is used for the nutrient medium containing copper nano-particle of LJ-king peppers.
Specific implementation mode
The iron of electroneutral, zinc and copper nano-particle use device Migen-3 (be recorded in Jigatch A.N., Leipunskii I.O.,Kuskov M.L.,Stoenko N.I.,Storozhev V.B."An apparatus for the production and study of metal nanoparticles",Instrum.Exp.Tech.43(2000)839- 845) it is prepared (AS SSSR № 814432, Bull.inventions.1981, № 11, С .25) by method described in document.
The estimation of Nanoparticle shape and size by using LEO 912AB OMEGA devices transmission electron microscope (TEM) method carries out.
The analysis of nano-particle phase composition by using ADP-1 diffractometers (Russia) X-ray diffraction analysis (XRD) method It carries out.
In order to determine the average diameter of nano-particle, TEM image handle by using 25 programs of computer Micran (with Based on measurement at least 1,000 particle diameters).Based on the data obtained, its point is drawn according to the size of metal nanoparticle Cloth curve, and calculate the average diameter of particle.
The TEM image and size distribution curve of metal nanoparticle are shown in Figure 1A -1C.The Size Distribution of Fe nanometer particles Curve is located in the region of 5-80nm.The average diameter of iron particle is 27.0 ± 0.51nm.The Size Distribution of zinc nano-particle is bent Line is located in the region of 10-200nm.The average diameter of zinc particles is 54.0 ± 2.8nm.The size distribution curve of copper nano-particle In the region of 5-225nm.The average diameter of copper particle is 79.0 ± 1.24nm.
According to Figure 1A -1C, Fe nanometer particles, zinc nano-particle and copper nano-particle are covered by translucent oxidation film Mono-crystalline structures with round regular shape.
XRD analysis is the result shows that the crystal content of Fe nanometer particles is as follows：Ferrous metal is equal to 53.6%, aoxidizes iron phase (Fe3O4) it is equal to 46.4%, the thickness of oxidation film is 3.5nm.Zinc nano-particle and copper nano-particle are by with 1.0-0.5nm The crystalline metal phase composition of thickness oxidation object film.
About the preparation of nutrient medium according to the present invention, any of balanced nutrients culture medium is (containing to plant Vital all organic and inorganic macronutrient and micronutrient) preparation can use based on.Therefore, it is Nutrient medium according to the present invention is prepared, electricity consumption neutrality Fe nanometer particles, electroneutral zinc nano-particle or electroneutral copper receives Rice corpuscles or combination thereof are introduced into instead of the combination of molysite, zinc salt or mantoquita or these salt in culture medium respectively, simultaneously Other components of nutrient medium as basis remain unchanged.
In order to prove the possibility of the invention implemented, agar Murashing-Skoog nutrient media formulations are used herein The nutrient medium composition according to embodiment of the present invention is prepared based on (referring to table 1).
In embodiments of the present invention, the selection of Fe nanometer particles, zinc nano-particle and copper nano-particle concentration range In view of based on culture medium intermediate ion form these metals content.Therefore, in terms of metal, basal MS medium A concentration of 5.6mg/L of middle iron ion, in embodiments of the present invention, the concentration of electroneutral Fe nanometer particles in nutrient medium For 10.0-0.06mg/L；A concentration of 1.96mg/L of zinc ion in basal MS medium in terms of metal, in the reality of the present invention It applies in mode, a concentration of 3.0-0.016mg/L of the electroneutral zinc nano-particle in nutrient medium；In basal MS medium with A concentration of 0.0064mg/L of the copper ion of metal meter, in embodiments of the present invention, the electroneutral copper in nutrient medium A concentration of 0.04-0.00016mg/L of nano-particle.
The preparation of basic MS nutrient mediums storing solution (control group) and the nutrient medium according to embodiment of the present invention Method is as described below.
1, MS nutrient mediums (control group) are prepared
MS nutrient mediums are prepared according to the formula that table 1 provides.
1.1, the stock solution of main salt (macronutrient) is prepared.
By the weighing section in terms of mg：NH4NO333000, KNO338000, CaCl2×2H2O 8800, MgSO4×7H2O 7400, KH2PO43400 are dissolved in 1 liter of water, are stirred 5 minutes on magnetic stirring apparatus " IKA RH basic 2 " (Germany).
1.2, the stock solution of trace salt (micronutrient) is prepared.
By the weighing section in terms of mg：KJ 166, H3BO31240, MnSO4×7H2O 4460, ZnSO4×7H2O 1720, Na2MoO4×2H2O 50, CuSO4×5H2О 5, CoCl2×6H2O 5 is dissolved in 1 liter of water, and 5 points are stirred on magnetic stirring apparatus Clock.
1.3, the stock solution of iron chelate is prepared.
By the weighing section in terms of mg：Na2EDTA×2H2O 37.3 and FeSO4×7H2O 27.8 is dissolved in 1 liter of water, It is stirred 5 minutes on magnetic stirring apparatus.
1.4, the stock solution of vitamin and organic matter is prepared.
By the weighing section in terms of mg：Inositol 100000, niacin (PP) -500, pyridoxol-HCl (B6) -500, thiamine - HCl(B1) 500, glycine 2000 is dissolved in 1 liter of water, is stirred 5 minutes on magnetic stirring apparatus.
Prepare the weighing section in terms of mg：The sucrose 30000 of powder type, agar 7000.
1.5, MS nutrient mediums are prepared.
In order to prepare 1L culture mediums, by the stock solution of 50mL macronutrients, the stock solution of 5mL micronutrients and The stock solution and the stock solution of 1mL vitamins and organic matter of 5mL chelated irons are dissolved in 800mL water, nuclear-magnetism blender Stirring 5 minutes.Then, the weighing section of sucrose and agar is added, and volume is adjusted to 1000mL.
In autoclave " in Hirayama, HVE-50 " (Japan), by nutrient medium in 120 DEG C of temperature, 0.1MPa It sterilizes 20 minutes under pressure.
As a control group by the Murashing-Skoog nutrient mediums of above-mentioned preparation.
2, the nutrient medium added with chitosan is prepared.
100mg chitosans Tyanshi (China) is added to the sterile MS nutrition training for being cooled to 45 DEG C of the above-mentioned preparations of 1L It supports in base, and mixed by stirring 2 minutes on magnetic stirring apparatus.
3, the nutrient medium containing metal nanoparticle is prepared.
3.1, chelating agent solution is prepared.
By weighing section 37.3mg Na2EDTA·2H2O is dissolved in 1L water, while being stirred 5 minutes on magnetic stirring apparatus.
3.2, the water slurry of Fe nanometer particles is prepared.
In order to prepare the deposit water slurry of Fe nanometer particles, the 2000mg powder for containing electroneutral Fe nanometer particles is put Enter in flask, and 200mL sterile distilled waters are added.Then, by " Scientz JY 92-IIN " disperser (China) with 99% Power by sample dispersion 30 seconds, the suspension is 30 seconds cooling in ice bath, and it is 10-15 DEG C to make temperature.The dispersion process weight Again three times.
By 9.0mL sterile distilled waters be added to 1.0mL preparation suspension in, and by above description carry out dispersion and it is cold But.
In order to prepare the Fe nanometer particles suspension of smaller concentration, the deposit Fe nanometer particles are suspended with sterile distilled water Liquid is diluted to required concentration, then carries out dispersion and cooling stage as described above.
3.3, the nutrient medium containing Fe nanometer particles is prepared.
The deposit water slurry of 10mL Fe nanometer particles or its dilution appropriate are added to 1L the 1st (referring to 3.2) Point description it is prepared it is sterile, be cooled in 45 DEG C of nutrient medium, but do not add iron chelate stock solution (referring to 1.3), then 5mL chelating agents are added to the stock solution that iron chelate is replaced in solution (referring to 3.1).The mixture is existed It is stirred 2 minutes on magnetic stirring apparatus.
3.4, the water slurry of zinc nano-particle is prepared.
In order to prepare the deposit water slurry of zinc nano-particle, the 600mg powder for containing electroneutral zinc nano-particle is put Enter in flask, and the distillation sterile water of 200mL is added.Then, by sample with 99% power dispersion 30 seconds, and suspension is existed 30 seconds cooling in ice bath, it is 10-15 DEG C to make temperature.The dispersion process is in triplicate.
9.0mL sterile distilled waters are added in the suspension of 1.0mL preparations, and according to described by iron powder carrying out Dispersion and cooling procedure (referring to 3.2).
In order to prepare the zinc nanoparticle suspension of smaller concentration, zinc nanoparticle suspension will be laid in sterile distilled water It is diluted to required concentration, then carries out dispersion and cooling stage as described above.
3.5, the nutrient medium containing zinc nano-particle is prepared.
The deposit water slurry of 10mL zinc nano-particles or its dilution appropriate are added to 1L according to (referring to 3.4) 1 part prepare it is sterile, be cooled in 45 DEG C of nutrient medium, but do not add zinc sulfate.By the mixture in magnetic agitation It is stirred 2 minutes on device.
3.6, the water slurry of copper nano-particle is prepared.
In order to prepare the deposit water slurry of copper nano-particle, the 8mg powder for containing electroneutral copper nano-particle is put into Flask, and 200mL sterile distilled waters are added.Then, by sample with 99% power dispersion 30 seconds, the suspension is cold in ice bath But 30 seconds, it was 10-15 DEG C to make temperature.The dispersion process is repeated 3 times.
9.0mL sterile distilled waters are added in the suspension of 1.0mL preparations, and according to described by iron powder carrying out Dispersion and cooling procedure (see 3.2).
In order to prepare the copper nano-particle suspension of smaller concentration, the deposit water of copper nano-particle is hanged with sterile distilled water Supernatant liquid is diluted to necessary concentration, then carries out dispersion and cooling stage by above description.
3.7, the nutrient medium containing copper nano-particle is prepared.
The deposit water slurry of 10mL copper nano-particles or its dilution appropriate are added to 1L according to (referring to 3.6) The description of 1 part prepare it is sterile, be cooled in 45 DEG C of nutrient medium, but do not add copper sulphate.By the mixture in magnetic It is stirred 2 minutes on power blender.
3.8, the water slurry containing iron, zinc and copper nano-particle is prepared.
It, will in order to prepare the water slurry of the Fe nanometer particles containing different proportion, zinc nano-particle and copper nano-particle The water slurry (referring to 3.2,3.4 and 3.6) of the required concentration of above-mentioned preparation is mixed with each 2mL, and by the sterile steaming of sample Distilled water is adjusted to 10mL, then according to described by Fe nanometer particles being disperseed above and cooled down (referring to 3.2).
3.9, the nutrient medium containing Fe nanometer particles, zinc nano-particle and copper nano-particle is prepared.
The metal nanoparticle suspension of the above-mentioned preparations of 10mL is added to 1L (referring to 3.8) and describes institute according to part 1 Prepare it is sterile, be cooled in 45 DEG C of nutrient medium, but do not add iron chelate (referring to 1.3), zinc sulfate (ZnSO4× 7H2) and copper sulphate (CuSO O4×5H2O stock solution).Then 5mL chelating agent solutions are added (referring to 3.1).By the mixture It is stirred 2 minutes on magnetic stirring apparatus.
The nutrient medium of the above-mentioned preparations of 50mL is dispensed into 200mL aseptic bottles immediately (referring to 3.3,3.5,3.7 and 3.9) In, it is sealed with polyethylene film or polypropylene cap, and stay in cooling in desinfection chamber.
The nutrient medium prepared according to described method is further used for germination and plant culture.
The nutrient medium sample containing metal nanoparticle added with a concentration of 100mg/L Tyanshi chitosans Preparation method describes similar added with the Murashing-Skoog nutrient mediums of chitosan with part 2.
The present invention realizes as follows：
The seed of plant is placed in sterile culture container, 3 seeds of each container, according to the present invention, agar nutrition training Contain the combination of Fe nanometer particles, zinc nano-particle or copper nano-particle or these nano-particle different proportions in the surface for supporting base.
The container is placed on the shelf of climatic chamber (22-25 DEG C, humidity 36%), with the daily luxs 3500-3000 It is illuminated within 12/12 hour.It is received using standard MS medium (control group) and using containing Tyanshi chitosans and/or metal The nutrient medium (test group) of rice corpuscles carries out independent every group of experiment.
Estimated the germinating capacity of seed at the 15th day, the form and physiological parameter of planting plants are the after plant culture starts It measures within 40 days.
Repeat 7-10 time obtain experimental result with 6.0 computer programs of Microsoft Excel Statistica into Row statistical disposition.As a result reliability (p) uses U- criterion evaluations (the Mann H.B., Whitney of Mann-Whitney D.R.Ann.Math.Statist.1947,V.18,№1,pp.50-60).Difference between two kinds of samples 0.001≤p≤ It is statistically significant when 0.1.
It is the reality that pepper and tomato plants are germinateed and grown on the nutrient medium according to embodiment of the present invention below Example is applied, the nutrient medium contains the Fe nanometer particles of various concentration, zinc nano-particle or copper nano-particle or their group It closes.These embodiments only illustrate the possibility that the present invention is implemented, but are not intended to limit the be possible to modification of its realization.
Embodiment 1：The influence that metal nanoparticle and chitosan in nutrient medium composition germinate to tomato seeds.
Germination is characterized with the numerical value of germination index, and germination index is defined as the quantity of chitting piece relative to institute Take the percentage of seed sum.Seed germination index measures on the 15th day after plant culture starts.
Fig. 2A -2D are shown in nutrient medium composition, and metal salt and chitosan are replaced with metal nanoparticle The influence germinateed to Venice tomato seeds of supplement (100mg/L).
Fig. 2A is shown on the nutrient medium of the Fe nanometer particles containing a concentration of 0.6-10.0mg/L, tomato seeds Germination index improves 13-27% (compared with the control group).Chitosan is added into the nutrient medium containing Fe nanometer particles, is shown Bursting of buds index additionally improves 23-33%.
As shown in Figure 2 B, compared with the control group, the zinc nanoparticle of a concentration of 0.2-3.0mg/L is supplemented in nutrient medium Son makes the germination index of seed increase 2-3 times.It is poly- that shell is added into the nutrient medium containing 1.0mg/L zinc nano-particles Sugar makes germination index further increase 13%.
As shown in Figure 2 C, compared with the control group, in nutrient medium a concentration of 0.0008-0.04mg/L copper nano-particle Tomato seeds germination is set to improve 2.0-2.8 times.It adds chitosan into containing a concentration of 0.0008mg/L's and 0.04mg/L In the nutrient medium of copper nano-particle, germination is made additionally to improve 20%.
As shown in Figure 2 D, compared with the control group, the mixture of Fe nanometer particles, zinc nano-particle and copper nano-particle is mixed Enter into nutrient medium to replace the salt of these metals, so that tomato seeds is germinateed is improved, in addition, nanometer in culture medium The concentration of particle is higher, and the germination index of tomato seeds is higher.For example, compared with the control group, combination：Fe nanometer particles (10.0mg/L)+zinc nano-particle (3.0mg/L)+copper nano-particle (0.04mg/L) makes the germination of tomato seeds improve to 2.7 Times.Chitosan is added to containing combination：Fe nanometer particles (3.0mg/L)+zinc nano-particle (1.0mg/L)+copper nano-particle (0.004mg/L)) nutrient medium in, cause the germination index of tomato seeds to further increase 13%.
Embodiment 2：The shadow of metal nanoparticle in nutrient medium composition to pepper and tomato plants morphological index It rings.
The morphological index (germination length, root long degree and green portion Biomass yield) of plant is received from plant containing metal It measures within the 40th day after culture starts (under these conditions) on the nutrient medium of rice corpuscles.As a result in experiment (test group) and Percentage size between the index obtained in control group indicates.
Fig. 3 A-3D show the metal nanoparticle in nutrient medium composition to the root long degree of LJ-king peppers It influences.
As can be seen that compared with the control group, containing a concentration of 0.06mg/L, 0.3mg/L and 3.0mg/L from Fig. 3 A Fe nanometer particles cultivate pepper on the nutrient medium instead of the iron (being calculated as 5.6mg/L with metal) of ionic species, lead to root long Degree has increased separately 54%, 118% and 102%.The iron of the concentration ratio ionic species of Fe nanometer particles almost low two in culture medium When a order of magnitude, this positive effect is observed.
Fig. 3 B are shown, compared with the control group, if pepper is containing a concentration of 0.4mg/L, 0.008mg/L and 0.016mg/L Zinc nanoparticle seed replace cultivating on the nutrient medium of zinc ion (being calculated as 1.96mg/L with metal), the root long degree of pepper point 71%, 80% and 62% is not increased.Therefore, the optium concentration of zinc nano-particle seems than the zinc ion in MS nutrient medium Concentration it is 5-122 times low.
Chitosan, which is added in the nutrient medium containing 0.4mg/L zinc nano-particles, causes root long degree to add additional 30%.
Fig. 3 C explanations contain a concentration of 0.00016mg/L, 0.0008mg/ respectively when pepper is planted in compared with the control group When the copper nano-particle of L and 0.004mg/L is replaced on the nutrient medium of copper ion (being calculated as 0.0064mg/L with metal), pepper Root long degree increase 34%, 57% and 54%.Therefore, in the concentration ratio MS nutrient mediums of copper nano-particle copper ion it is dense When spending low 1.6-40 times, positive effect is observed.
Fig. 3 D are shown, compared with the control group, when in the Fe nano-particles containing different proportion, Zn nano-particles and Cu nanometers When the combination of particle on the nutrient medium of iron ion, zinc ion and copper ion instead of cultivating pepper, the root long degree of pepper increases 7-58%.Wherein nano-particle is the Cmin (iron 0.06mg/L, zinc 0.016mg/L and copper 0.00016mg/L) of test group Experiment be exception.
But Fig. 3 D show that low-concentration metallic nano-particle and chitosan collective effect have significant synergistic effect, cause The root long degree of pepper increases 40% compared with the control group.
Containing chitosan and combination：Fe nanometer particles (0.3mg/L)+zinc nano-particle (0.08mg/L)+copper nanoparticle Similar synergistic effect is also obtained in the case of the nutrient medium of sub (0.0008mg/L).
As explanation, Fig. 4 shows the germination photo for the LJ-king peppers being planted on nutrient medium, wherein nutrition Zinc salt in culture medium is replaced by the combination of chitosan and zinc nano-particle.It is obvious that compared with the control group, in nutrient medium In there are a concentration of 0.016-0.4mg/L zinc nano-particles and chitosan, and pepper root system to be contributed to more energetically to develop：Root long degree Increase 1.6-1.8 times.Chitosan is added in the nutrient medium of the zinc nano-particle containing a concentration of 0.4mg/L, is made Root long degree adds additional 30%.
Fig. 5 A-5D show metal nanoparticle in nutrient medium composition and chitosan to HY-2 tomato root length Influence.It is planted in the battalion that ionic species iron is replaced containing a concentration of 0.06mg/L, 0.3mg/L and 3.0mg/L Fe nanometer particles Tomato in foster culture medium shows root long degree and has increased separately 58%, 31% and 3% (referring to Fig. 5 A).
When tomato planting respectively contain a concentration of 0.016mg/L, 0.08mg/L and 0.4mg/L zinc nano-particle replace from When in the nutrient medium of sub- form zinc, root long degree has increased separately 1%, 17% and 38%.With the supplement of chitosan, the knot Fruit has increased separately 17%, 2% and 15% (referring to Fig. 5 B).
The copper ion of a concentration of 0.0064mg/L is substituted with the copper nano-particle of a concentration of 0.0008mg/L leads to tomato Root long degree increases 24% (referring to Fig. 5 C).
When tomato planting is in the combination (0.3mg/L iron+0.08mg/L zinc+0.0008mg/L copper) containing metal nanoparticle (3.0mg/L iron+0.4mg/L zinc+0.004mg/L copper) and when in the nutrient medium of non-ionic form metal, with control group It compares, the root long degree of tomato has increased separately 7% and 10%.In some experiments, the addition of chitosan promotes tomato root long Degree further increases.For example, compared with the control group, Fe nanometer particles (3.0mg/L), zinc are applied in combination in nutrient medium Nano-particle (0.4mg/L), copper nano-particle (0.004mg/L) and chitosan make the root long degree of tomato increase 70%.
Include germination length and green portion biomass of the metal nanoparticle to pepper and tomato plants in the medium With actively impact.Therefore, zinc nano-particle is introduced into the concentration of 0.08mg/L in nutrient medium and HY-2 tomatoes is promoted to send out Bud length increases 1.2 times.Chitosan adds in nutrient medium and the index is further increased 17%.
Fe nanometer particles, which are introduced into the concentration of 3.0mg/L in nutrient medium, promotes LJ-king peppers germination length to increase Add 4.7%.Chitosan, which adds in nutrient medium, further makes the index increase 8.9%.
Zinc nano-particle, which is introduced into the concentration of 0.2mg/L in nutrient medium, promotes Venice tomatoes germination length to increase 15.5%.
It is planted in containing the green of the LJ-king peppers on zinc nano-particle (0.4mg/L) and the nutrient medium of chitosan Color part biomass in control group than being higher by 42%.Compared with the control group, using the zinc nanoparticle containing a concentration of 0.08mg/L The nutrient medium of son makes its green portion biomass increase 80% for planting Venice tomatoes.Group containing nano-particle Closing the nutrient medium of (3.0mg/L iron+0.4mg/L zinc+0.004mg/L copper) makes the green portion biology volume production of Venice tomatoes Amount is higher by 1.3 times than control group.
Embodiment 3：The shadow of metal nanoparticle in nutrient medium composition to pepper and tomato plants physical signs It rings.
The physical signs (chlorophyll content of root activity and blade) of planting plants cultivates (under these conditions) from plant It records within the 40th day after beginning.
The result is indicated with the percentage for testing exponential quantity and control group exponential quantity.Test value is from containing metal nanoparticle Nutrient medium obtain, control value obtains from Murashing-Skoog nutrient mediums.
Root activity by described chlorination 3- phenyltetrazoles it is also original measurement [Adebusoye O.Onanuga, Ping’an Jiang,Sina Adl."Effect of phytohormones,phosphorus and potassium on cotton varieties(Gossypium hirsutum)root growth and root activity grown in hydroponic nutrient solution",Journal of Agricultural Science.2012,Vol.4,N 3, pp.93-110]。
Fig. 6 A-6D, which show to be introduced into nutrient medium, replaces the metal nanoparticle of corresponding metal salt to LJ- King peppers and the active influence of Venice tomato roots.
It will be apparent that, trained respectively in the nutrition containing a concentration of 0.06mg/L and the Fe nanometer particles of 0.3mg/L from Fig. 6 A When supporting culture pepper on base, the root activity of LJ-king peppers improves 59% and 58%.Compared with the control group, containing concentration The HY-2 tomato plants cultivated in nutrient medium for the Fe nanometer particles of 0.06mg/L, 0.3mg/L and 3.0mg/L, root Activity has been respectively increased 37%, 34% and 48%.
Compared with the control group, in the culture of the Fe nanometer particles containing a concentration of 0.6mg/L, 3.0mg/L and 10.0mg/L Venice tomato plants are cultivated in base, its root activity is caused to be respectively increased 112%, 125% and 76%.
When replacing the zinc ion in nutrient medium with zinc nano-particle, observe similar effect (referring to Fig. 6 B).With Control group is compared, and LJ-king pepper plants are planted in the zinc nanometer containing a concentration of 0.016mg/L, 0.08mg/L and 0.4mg/L When in the nutrient medium of particle, root activity has been respectively increased 31%, 56% and 38%.In these cases, with control group It compares, the root activity of HY-2 tomatoes has been respectively increased 86%, 8% and 29%.
Compared with the control group, Venice tomato plants are planted in containing a concentration of 0.2mg/L, 1.0mg/L and 3.0mg/L When in nutrient medium, root activity has been respectively increased 25%, 15% and 42%.
Fig. 6 C show that the root activity of the pepper being planted on the nutrient medium containing copper nano-particle and tomato plants carries It is high.
Copper ion is replaced in the copper nano-particle containing a concentration of 0.00016mg/L, 0.0008mg/L and 0.004mg/L Pepper is cultivated on nutrient medium, and the root activity of LJ-king peppers is caused to be respectively increased 18%, 61% and 21%.With compare Group is compared, and the root activity of HY-2 tomatoes has been respectively increased 43%, 151% and 149%.
Compared with the control group, in the copper nano-particle containing a concentration of 0.0008mg/L, 0.004mg/L and 0.04mg/L Venice tomatoes are cultivated on nutrient medium, its root activity is made to be respectively increased 65%, 10% and 12%.
Most of experiments show in nutrient medium, combination replacement iron ion, the zinc ion of these metal nanoparticles And copper ion, it improves to the active actively impact of root (referring to Fig. 6 D).
In the combination containing nano-particle：(0.06mg/L iron+0.016mg/L zinc+0.00016mg/L copper) and (0.3mg/L The copper of the zinc+0.0008mg/L of iron+0.08mg/L) and (3.0mg/L iron+0.4mg/L zinc+0.004mg/L copper) replace ion shape Pepper is cultivated in the nutrient medium of these metals of formula, the root activity of pepper is caused to be respectively increased 98%, 51% and 91%.
Compared with the control group, in the combination containing nano-particle：(0.06mg/L iron+0.016mg/L zinc+0.00016mg/L Copper) and (0.3mg/L iron+0.08mg/L zinc+0.0008mg/L copper) replace ionic species these metals nutrient medium in HY-2 tomatoes are cultivated, cause the root activity of HY-2 tomatoes that 33% and 46% has been respectively increased.
Compared with the control group, in the combination containing nano-particle：(0.6mg/L iron+0.2mg/L zinc+0.0008mg/L copper) It is cultivated in the nutrient medium of (3.0mg/L iron+1.0mg/L zinc+0.004mg/L copper) instead of these metals of ionic species Venice tomatoes cause the root activity of Venice tomatoes that 43% and 48% has been respectively increased.
In most cases, chitosan is added to and promotes plant in the nutrient medium containing metal nanoparticle Root activity is additional to be improved.
The chlorophyll content of blade according to described is assessed (" Measurement and Characterization by UV-VIS Spectroscopy UNIT F4.3",Current Protocols in Food Analytical Chemistry,2001,F4.3.1-F4.3.8).The result received is indicated by Fig. 7 A-7C.
Fig. 7 A, which show Fe nanometer particles being added in nutrient medium, replaces ferrous sulfate to LJ-king pepper plants Blade chlorophyll content influence.It has been shown that compared with the control group, a concentration of 0.3mg/L and 3.0mg/L in culture medium Fe nanometer particles make chlorophyll content increase separately 5% and 27%.Chitosan is added to nutrient medium, and it is a concentration of 0.06mg/L and 0.3mg/L Fe nanometer particles cause the chlorophyll content of pepper blade to further increase 6% He respectively together 10%.
Fig. 7 B, which show zinc nano-particle being added in nutrient medium, replaces zinc sulfate to the HY-2 tomato leaf middle periods The influence of chlorophyll contents.The zinc nano-particle of a concentration of 0.08mg/L and 0.4mg/L makes the leaf of HY-2 tomato leafs in culture medium Chlorophyll contents have increased separately 56% and 108%.Chitosan is additionally introduced in nutrient medium, if zinc nano-particle is dense Degree is 0.016mg/L and 0.08mg/L, which has increased separately 48% and 20%.
Fig. 7 C, which show to add to copper nano-particle in nutrient medium, replaces copper sulphate in LJ-king pepper blades The influence of chlorophyll content.Copper nano-particle is added in nutrient medium, and the chlorophyll for increasing LJ-king pepper blades contains Amount.Observe that maximum efficiency, the content of Determination of Chlorophyll are higher than in control group at the minimum test concentrations 0.00016mg/L of copper Go out 59%.
Chitosan is introduced in the nutrient medium containing 0.0008mg/L copper nano-particles, the chlorophyll of pepper blade contains Amount increases 75%.
It needs it is important to note that ground is, in nutrient medium, by the electroneutral of the iron of ionic species, zinc and copper these metals Metal nanoparticle replaces, and will not cause any interference to the development of plant：Plant is kept upright, and has well-developed impeller (leaf plates) and maintain blade replacement and other species characteristic features.In addition, plant is to bacterium, mould and fungi Infection has resistance.
It is therefore desirable to protect the plant cover cultivation methods based on using nutrient medium, wherein in nutrient medium The salt of iron, zinc and copper is partially or completely replaced by the electroneutral nano-particle of these metals.This method can be produced with densification The health plant of stem, the plant have flourishing and active root system, may be used as the planting material of high quality.
The nutrient medium proposed can be used for biotechnology research, the quality for improving agricultural product, be planted for aerosol Training technology and water culture technique.The method of planting plants on the nutrient medium containing iron, copper and zinc nano-particle proposed It can be used to create life support system under conditions of long-term space flight for astronaut.

Claims (10)

1. a kind of cultural method of plant, this method include：Aseptically, in the agar nutrition culture containing nano-particle Make germination on base and then cultivate plant, wherein the nano-particle is that electroneutral Fe nanometer particles or electroneutral zinc are received Rice corpuscles or electroneutral copper nano-particle or combination thereof.
2. according to the method described in claim 1, wherein, the nutrient medium also contains chitosan.
3. according to the method described in claim 1 and 2, wherein the nutrient medium contains a concentration of 10.0-0.06mg/L's Electroneutral Fe nanometer particles.
4. according to the method described in claim 1 and 2, wherein the nutrient medium contains a concentration of 3.0-0.016mg/L's Electroneutral zinc nano-particle.
5. according to the method described in claim 1 and 2, wherein the nutrient medium contains the electricity of 0.04-0.00016mg/L Neutral copper nano-particle.
6. the nutrient medium for implementing method according to claim 1, which, which contains, is included in Ingredient in Murashing-Skoog culture media compositions：Vitamin Р Р, 6 В and В 1, glycine, sucrose, meso flesh Alcohol, agar, chelating agent Na2EDTA×2H2O, inorganic salts NH4NO3、KNO3、CaCl2×2H2O、MgSO4×7H2O、KH2PO4、KI、 H3BO3、MnSO4×7H2O、NaMoO4×2H2O and CoCl2×6H2O, Yi Jitie, zinc and copper, wherein iron, zinc or copper or combinations thereof The nutrition culture is included in the form of Fe nanometer particles, zinc nano-particle or copper nano-particle or combination thereof respectively In base.
7. nutrient medium according to claim 6, wherein the nutrient medium also contains chitosan.
8. according to the nutrient medium described in claim 6 and 7, wherein the nutrient medium contains a concentration of 10.0- The electroneutral Fe nanometer particles of 0.06mg/L.
9. according to the nutrient medium described in claim 6 and 7, wherein the nutrient medium contains a concentration of 3.0- The electroneutral zinc nano-particle of 0.016mg/L.
10. according to the nutrient medium described in claim 6 and 7, wherein the nutrient medium contains a concentration of 0.04- The electroneutral copper nano-particle of 0.00016mg/L.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113575313A (en) * 2021-07-28 2021-11-02 江南大学 Application of copper nano fertilizer in improvement of tomato resistance and yield

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107646618A (en) * 2017-10-20 2018-02-02 成都盛世光华生物科技有限公司 A kind of plant nutrition liquid for meeting soilless culture cuckoo and preparation method thereof
US11483988B2 (en) 2017-11-30 2022-11-01 OnePointOne, Inc. Vertical farming systems and methods
RU2698718C1 (en) * 2018-12-26 2019-08-29 Федеральное государственное бюджетное учреждение "Национальный исследовательский центр "Курчатовский институт" Method for quantitative determination of weight of carbon nanostructures in samples
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RU2757791C1 (en) * 2021-03-10 2021-10-21 Ооо "Мещерский Научно-Технический Центр" Method for pre-sowing treatment of spring barley seeds on gray forest soils using nanoparticles of copper and copper oxide
CN113287634B (en) * 2021-06-09 2022-02-01 江南大学 Method for controlling tomato wilt based on nano elemental sulfur

Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH05123070A (en) * 1991-07-10 1993-05-21 Naasarii Technol:Kk Selection of young plant obtained by tissue culture
CN101142893A (en) * 2007-10-09 2008-03-19 浙江大学 Water-planting method for arabidopsis thaliana
CN101548644A (en) * 2009-03-25 2009-10-07 刘星华 Tissue culturing method for applying nanotechnology to promote fast proliferation of dendrobium officinale
CN102257942A (en) * 2010-05-27 2011-11-30 盐城中绿农业发展有限公司 Nano-selenium-enriched grape and cultivation method thereof
CN102643128A (en) * 2012-04-09 2012-08-22 湖北圣峰药业有限公司 Nano-selenium compound fertilizer
CN103430719A (en) * 2013-07-24 2013-12-11 王运海 Method for planting dangshen
CN103858685A (en) * 2014-03-13 2014-06-18 贵州中烟工业有限责任公司 Method for reducing lead content of tobacco leaves
CN104170558A (en) * 2014-08-12 2014-12-03 安徽永兴岛生态农业科技有限公司 Salvia miltiorrhiza planting method
CN104277983A (en) * 2014-09-17 2015-01-14 浙江中医药大学 Culture medium for fungal culture and preparation method of culture medium
WO2015020542A1 (en) * 2013-08-08 2015-02-12 Extenday Ip Limited Ground cover materials
CN104823736A (en) * 2015-04-07 2015-08-12 上海应用技术学院 Root control fabric used for container cultivation, and preparation method and application thereof
CN104838841A (en) * 2015-04-20 2015-08-19 天津师范大学 Method adopting salt-tolerant reinforcement nanometer garbage compost for reinforcing turf grass salt resistance
CN105272396A (en) * 2015-10-08 2016-01-27 武汉纺织大学 Composting treatment method of excess sludge from urban sewage treatment plant
CN105347983A (en) * 2015-12-23 2016-02-24 山东农业大学 Special super-large-granule humic acid fertilizer for fruit trees and preparation method of special super-large-granule humic acid fertilizer

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100450364C (en) * 2002-01-15 2009-01-14 崔光洙 The liquid composition for promoting plant growth, which includes nano-particle titanium dioxide
UA40794U (en) * 2008-11-24 2009-04-27 Владимир Борисович Борисевич Method for activation of microbiological processes, acceleration of growth and multiplication of microorganisms complex biophysical-and biochemical effect in microbiological processes
WO2011059507A1 (en) * 2009-11-13 2011-05-19 Biris Alexandru S Method of using carbon nanotubes to affect seed germination and plant growth
UA56207U (en) * 2010-05-25 2011-01-10 Николай Васильевич Косинов Method for microclonal propagation of grape in vitro
KR101478564B1 (en) * 2013-05-30 2015-01-06 주식회사 지앤에스바이오 Method of cultivating crops

Patent Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH05123070A (en) * 1991-07-10 1993-05-21 Naasarii Technol:Kk Selection of young plant obtained by tissue culture
CN101142893A (en) * 2007-10-09 2008-03-19 浙江大学 Water-planting method for arabidopsis thaliana
CN101548644A (en) * 2009-03-25 2009-10-07 刘星华 Tissue culturing method for applying nanotechnology to promote fast proliferation of dendrobium officinale
CN102257942A (en) * 2010-05-27 2011-11-30 盐城中绿农业发展有限公司 Nano-selenium-enriched grape and cultivation method thereof
CN102643128A (en) * 2012-04-09 2012-08-22 湖北圣峰药业有限公司 Nano-selenium compound fertilizer
CN103430719A (en) * 2013-07-24 2013-12-11 王运海 Method for planting dangshen
WO2015020542A1 (en) * 2013-08-08 2015-02-12 Extenday Ip Limited Ground cover materials
CN103858685A (en) * 2014-03-13 2014-06-18 贵州中烟工业有限责任公司 Method for reducing lead content of tobacco leaves
CN104170558A (en) * 2014-08-12 2014-12-03 安徽永兴岛生态农业科技有限公司 Salvia miltiorrhiza planting method
CN104277983A (en) * 2014-09-17 2015-01-14 浙江中医药大学 Culture medium for fungal culture and preparation method of culture medium
CN104823736A (en) * 2015-04-07 2015-08-12 上海应用技术学院 Root control fabric used for container cultivation, and preparation method and application thereof
CN104838841A (en) * 2015-04-20 2015-08-19 天津师范大学 Method adopting salt-tolerant reinforcement nanometer garbage compost for reinforcing turf grass salt resistance
CN105272396A (en) * 2015-10-08 2016-01-27 武汉纺织大学 Composting treatment method of excess sludge from urban sewage treatment plant
CN105347983A (en) * 2015-12-23 2016-02-24 山东农业大学 Special super-large-granule humic acid fertilizer for fruit trees and preparation method of special super-large-granule humic acid fertilizer

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
刘秀梅等: "纳米氧化铁对花生生长发育及养分吸收影响的研究", 《植物营养与肥料学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113575313A (en) * 2021-07-28 2021-11-02 江南大学 Application of copper nano fertilizer in improvement of tomato resistance and yield

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