CN104838841A - Method adopting salt-tolerant reinforcement nanometer garbage compost for reinforcing turf grass salt resistance - Google Patents
Method adopting salt-tolerant reinforcement nanometer garbage compost for reinforcing turf grass salt resistance Download PDFInfo
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Abstract
The invention discloses a method adopting salt-tolerant reinforcement nanometer garbage compost for reinforcing turf grass salt resistance. Salt-tolerant microbes are extracted from compost, pure salt-tolerant microbes are obtained through screening, a method for gradually improving NaCl concentration is adopted for further reinforcing and refining a reinforcement salt-tolerant bacterium agent, and the salt-tolerant bacterium agent and nanometer garbage compost are jointly applied to a saline-alkali soil turf medium. Three kinds of mild, medium and server salt stress treatment are designed by the experiment, reinforcement microbes are combined with nanometer garbage compost to reinforce the salt resistance of turf grass so as to provide basis for development of an effective fertilizer, improvement of salt resistance of turf grass and saline-alkali soil property and promotion of turf establishment in a salt stress environment.
Description
Technical field
The invention belongs to environmental protection technical field, relate to a kind of method adopting salt to strengthen active nano garbage compost enhancing Salinity Tolerance of Turfgrass.
Background technology
Nanoscale science and technology (Nano-ST) is the new science and technology emerged phase late 1980s, and its main research structure yardstick is (10
-7-10
-9m) character of material and application thereof in scope.Because nano material has the essential characteristics such as interface surface interfacial effect, small-size effect, macro quanta tunnel effect and quantum size effect, so, there is the Strange properties that many traditional materials do not possess.Nano material all shows the characteristic being different from traditional material in absorption, catalysis, magnetic effect and sensitivity characteristic, can with multiple fields height intersection, comprise physics, chemistry, electronics, material science and biology, high-tech application demonstrates vast potentiality, so applications to nanostructures has more and more received the concern of scientist.The Chinese government attaches great importance to the research of nanometer technology, and nanostructured and nano material is considered as the key of nanometer technology and is preferentially supported.2012, Chinese industrial and information-based portion are according to the spirit of " country's " 12 " emerging strategic industries development plan ", issue " new material industry " 12 " development plan ", nano material is classified as forward position field of new, and explicitly point out: China will strengthen nanotechnology research, promote the research and utilization of nano material in fields such as new forms of energy, environmental improvement, energy-saving and emission-reduction and bio-medicals energetically.
The external research for nano material mainly concentrates on the research to safety of nano materials, and the Five problems of main research is: the absorption of skin to nano material and the injury to skin thereof; Drink the consequence containing nano particle water; Nano particle is on the impact of animal lung; Become sedimental nano particle in water and affected 1 to 4 May in 2005 to surrounding environment, the European Community holds " meeting of European nano biological effect " at Bonn, Germany inside.The session discussing formation of nano particle, release, motion in the environment and detection method, the impact of its acute virus action, chronic toxic action and cellular level and molecular level, as the impact etc. on haemocyte, lung cells, immunocyte, DNA damage, gene expression.Domesticly there have been developed the impact of nano material on plant, nano fertilizer is proposed by soil and fertilizer research institute of the Chinese Academy of Agricultural Sciences Zhang Fudao researcher, and sets up the project in national " 863 " project.Nano fertilizer is a branch of nano biological science and technology, is the brand-new fertilizer building modification by nano material technology and medical microcapsules technology and make, is divided into nano material coating, cementing sustained-release and controlled release fertilizer and nanostructured fertilizer.As everyone knows, the groundwater contamination that abuse fertilizer causes is more and more serious, how to improve utilization rate of fertilizer and become the emphasis of dealing with problems, nano-material surface atom number accounts for more than 80% of complete particle total atom number, because lack adjacent atom around surface atom, containing many dangling bonds, so have unsaturation, be easy to combine with other atoms and settle out, thus show very high chemism.Can significantly improve the availability of fertilizer with Fertilizer combination, be the magnetic effect due to nano material on the one hand, can promote that nutrient is absorbed by plants, stimulating plant grows, and can also improve the activity of multiple enzyme in plant corpus.On the other hand, be because its skin effect, nano fertilizer surface energy and Surface binding energy are increased, and help it to be absorbed by root system of plant in soil environment, the effect of Fertilizer application is improved.There are some researches show, add the decline that nano-sized carbon fertilizer synergist effectively can slow down the available nutrient of soil, improve rice yield, amount of increase in production asking at 1.9%-3.5%.Nano-sized carbon, on the impact of rice yield and utilization rate of nitrogen fertilizer, also demonstrates and uses nano-carbon powder and can significantly improve rice yield, meanwhile, under optimum nitrogen rate condition, be affixed by the raising that nano-carbon powder contributes to nitrogen fertilizer for paddy rice availability.Nano material adhesive coating type slowly/controlled releasing fertilizer is on the impact of crop yield and quality, and result shows, nano material adhesive coating type slowly/controlled releasing fertilizer all contributes to the raising of wheat and maize seed output and protein output.The nano fertilizer majority of current research is nano material synergy fertilizer or nano material coat fertilizer, and fertilizer itself is worked into the research that nanometer particle size is applied to lawn plant environment stress, there is no bibliographical information.
Microbial manure is also called bacterial manure, bio-fertilizer, and Main Function shows and helps crop alimentary absorption, lifting chemical fertilizer utilization ratio and the aspect such as quality of agricultural product, enhancing crop anti-adversity.Microbial manure generally can be divided into two classes, and a class is the vital movement of microorganism by self, and increasing can for the nutritive element of plant absorption; Another kind of, by the secondary metabolism secreting hormone of microbial life activity, the resistance of Promoting plant growth and raising plant can be carried out.As nitrogen-fixing microorganism can increase Soil Nitrogen Content by nitrogen fixation, microorganism can improve conversion and the absorption of Nutrient Elements in Soil by vital movement, be the major function of most of microbe fertilizer.In addition, some beneficial microbes can produce various saccharides material by the metabolic activity of self, these glucides can not only organically combine with the gelatin substance in soil, and the mucus that simultaneously can also produce with plant self combines, and play the effect of improvement soil.Microbial bacterial agent also has and improves crop growth conditions, strengthens the effects such as plant resistance to environment stress.Current great majority research shows, the absorption of microorganism formulation energy Promoting plant growth and nutritive element.Have researcher to be applied to rice research, result shows, compared with conventional fertilizer application, applies the increase that bio-bacterial manure effectively can promote rice yield.Also have people the culture studies that the seven Plants growth-promoting fungies filtered out are applied to sunflower to be sent out, result proves, fungi can obviously promote its primary growth, and effectively can suppress the generation of downy mildew.In sum, the research of current related microorganism lays particular emphasis on beneficial microorganism more and extracts, identifies and application, but the research being applied to lawn plant adverse growth after being strengthened further by beneficial microbe there is no bibliographical information.
Turf establishment system integrates ecology regulation and control, beautifies the environment and the ecological engineering of the several functions such as entertainment.As the important component part of urban landscaping, green grassland is one of standard weighing economic development level, also be one of standard weighing modern city, be that modern city social development and economic strength one embodies effectively, the structure of turf establishment system is for improving City Investment Environment, inviting outside investment, promoting that the various aspects such as tourism all play a part can not ignore.Lawn, while bringing view and landscaping effect, also plays an important role maintaining in ecosystem balance.First, it can purify air, sterilizing, and lawn absorbs CO by photosynthesis
2release O
2keep CO in air
2and O
2balance, lawn can also be diluted, decompose, absorb and the noxious material such as toxic gas, carcinogen and some heavy metal gas in fixing air, is turned bane into boon by photosynthesis.Secondly, lawn can conserve water and soil, maintain the ecological balance.The fine and close grass layer covering earth's surface of lawn energy, and intensive root system can effectively set table be native, prevents the formation of rainwash, the possibility that reduction table soil is pulled away.In addition, solar radiation can also be slowed down in lawn, effectively reduces surface temperature, increases air humidity.
Saline land (soil) is salinization soil, the general name of alkali soil and saline-alkali soil.In recent years, the salinization of soil constantly increases the weight of.At present, there is 20 × 106hm in China
2above saline land and 7 × 106hm
2above salinization soil, accounts for 20% of arable area.No matter be in arid area, or in the coastal area that water resource is enriched, salt stress is all a kind of very serious abiotic stress.And salt stress is one of critical environments factor for restriction Turfgrass Growth.A large amount of salinization soils is fallen into disuse, and greatly wastes soil resource, hinders agricultural production and urban afforestation process.The harm main manifestations of salinity to crop is, percentage of seedgermination reduces, Developing restraint slows down, root growth slows down, yellow leaf, injury plant tissue, cause the phenomenons such as plant cell membrane damage, causes growing of plant to be severely limited.For a long time, what the improvement of saline-alkali soil mainly adopted is irrigation and drainage, the plant applying chemical improvement agent or mineral fertilizer and plantation Salt And Alkali Tolerance.For above problem, current research mainly concentrate on choose strong stress resistance lawn plant, use concentrated fertilizer, use the measure such as water-loss reducer and modifying agent.Nano fertilizer, as a kind of novel concentrated fertilizer, receives increasing concern in Adversity-stressed Plant.
Nano fertilizer is a kind of special concentrated fertilizer, the surface area of its particle, surface energy and Surface binding energy are all relatively large, contribute to it by the root hair of plant or tip of a root partial adsorbates in soil environment, its characteristic property likely solves the problems such as utilization rate of fertilizer, crop yield and plant resistance to environment stress.Research majority at present about nano fertilizer is nano material synergy fertilizer or nano material coat fertilizer, and fertilizer itself is worked into the research that nanometer particle size is applied to lawn plant, equally also has no correlation technique report.
Consumer garbage compost is one and highly oozes system, and microorganism wherein has certain adaptive capacity to adverse circumstance, will wherein beneficial microbe microbial inoculum to be applied to lawn adverse circumstance planting after strengthening further significant.Based on above consideration, in order to study nanometer compost and the effect of beneficial microbe in turf establishment thereof, garbage compost is worked into about 30nm through nanofabrication technique herein, and separation and Extraction goes out suitable beneficial microbe strain from consumer garbage compost, tamed by further resistance, obtain more efficient enhancement microbiological microbial inoculum, after being pressed certain volume ratio mixing, make complex micro organism fungicide, the turf establishment of to coerce etc. under environment stress is united and applied in nanometer compost, by exploring its impact on lawn plant physiological and ecological characteristic under environment stress, to being the application of enhancement microbiological associating nanometer compost in adverse circumstance turf establishment, promote that the planting of adverse circumstance environment lawn plant provides theoretical foundation.
Microorganism energy significantly Promoting plant growth, promotes the absorption of nutritive element.In addition, fungi effectively can improve the resistance such as cold resistance, drought resistance, opposing biohazard of lawn plant.Some results of study show, nanometer compost has obvious facilitation to plant growth, but technology nanometer compost being used for the aspect of salt stress turf establishment there is no bibliographical information.
Summary of the invention
The present invention extracts salt-durable microbe from compost, and screening is obtained purebred salt-durable microbe, adopts the concentration method progressively improving NaCl to strengthen white silk further and makes strengthening salt tolerant microbial inoculum, co-administered in saline-alkali soil lawn matrix with nanometer compost.Experimental design is slight, moderate and severe three kinds of salt stress process, realize the salt-resistance that enhancement microbiological associating nanometer compost strengthens turfgrass, object is development concentrated fertilizer, improves the salt-resistance of turfgrass, improvement saline-alkali soil character, promotes that the planting of salt stress environment lawn plant provides foundation.
The invention discloses following content for achieving the above object:
Adopt salt to strengthen a method for active nano garbage compost enhancing Salinity Tolerance of Turfgrass, it is characterized in that being undertaken by following step:
(1) preparation of nanometer compost:
Consumer garbage compost 105 DEG C is dried to constant weight, garbage compost processing grinding is made 30-35nm nanometer compost, for subsequent use;
(2) preparation of Salt-tolerant microbial agent:
Compost sample is taken 10g and be placed in aseptic conical flask, after adding 100mL sterile water shaken well, get 10mL suspension in the conical flask filling 100mL enriched medium, at 28 DEG C, shaken cultivation 3d under 220r/min, is mixed microorganism flora; Containing bacillus subtilis, lysine bacillus and penicillium chrysogenum in described mixed microorganism flora;
(3) what the strengthening of mixed microorganism flora salt tolerant adopted is the method progressively increasing salinity, to alleviate the moment impact that causes hybrid bacterial strain of high salt concentration and murder by poisoning, the concentration of NaCl is increased to 3% with the gradient of 0.5%, often increase the concentration of a NaCl, after thalline increment is stable, the concentration increasing NaCl could be continued, tame out salt-durable microbe flora step by step;
(4) preparation of compound Salt-tolerant microbial agent:
The proportions of bacillus subtilis, lysine bacillus and penicillium chrysogenum 1:1:1 by volume becomes compound Salt-tolerant microbial agent, dilute 100 times for subsequent use;
(5) turf established by sod:
Soil and saline-alkali soil to be sieved removal of impurities, make 0.3% slight (W/W), 0.6% moderate (W/W) and 0.9% severe (W/W) saline-alkali soil by a certain percentage, by for subsequent use for saline-alkali soil sterilizing 30 min at 121 DEG C mixed, each coercing establishes 3 process, three to coerce set process identical, is respectively:
(1) with 100% soil for contrast matrix add 15 ml blank cultures;
(2) soil and nanometer compost mix in the ratio of 59:1 and add 15ml blank cultures;
(3) soil and nanometer compost mix in the ratio of 59:1 and add 15ml strengthening Salt-tolerant microbial agent;
With the soil mixed for matrix carries out lawn plant experiment in cultivation, during experiment, substrates quantity is 9400gm
-2, compounded turf matrix thickness is 15 mm, and Festuca Arundinacea grass seeds seeding quantity is 160gm
-2, son is evenly sowed in matrix top layer after soaking 24h, first carries out dark processing, and fully water and ensure to sprout, cultivate after sprouting, intensity of illumination is 450-700 μm of ol.m
-2.s
-1period regularly waters a water sooner or later, the position that turf is cultivated in frequent exchange is consistent to ensure illumination, indoor relative moisture is 40-55%, temperature is 20-25 DEG C, and carry out the mensuration of each index of plant after 50d is carried out in experiment, the wherein said method progressively increasing salinity refers to: the concentration of NaCl is respectively 0.5%, 1%, 1.5%, 2%, 2.5% and 3%, in domestication process, depending on OD
600increase and progressively improve salinity, the concentration of NaCl is increased to 3% with the gradient of 0.5%.
The present invention further discloses the method adopting salt strengthening active nano garbage compost to strengthen Salinity Tolerance of Turfgrass and improve the application in plant root growth.Particularly in the chlorophyllous increase of promotion Festuca Arundinacea, improve the application of Festuca Arundinacea photosynthetic rate aspect.
The more detailed description of the present invention:
1 development materials and methods
1.1 experiment material
Choose full seed, perennial Festuca Arundinacea of uniform size (
festuca arundinaceal.) seed is test material.
Consumer garbage compost is from Tianjin little Dian composting plant, basic physical and chemical is: pH 7.62, saturation moisture content 0.76 ml/g, unit weight 0.85 g/ml, full nitrogen 5.18%, full potassium 50.83 g/kg, available phosphorus 77.92 mg/kg, organic 12.12%, garbage compost is removed the foreign material such as wood, plastics, metal wherein, air-dry rear for subsequent use.
Soil matrix is taken from school institute of Tianjin Normal University, impurities removing, and cross 2mm sieve, be positioned over ventilating opening place 2-3d, air-dry under natural conditions, its physicochemical property is: pH7.44, saturation moisture content 0.58mLg
-1, organic 4.68%, full nitrogen 0.21%, salt content 0.29%.
Saline-alkali soil takes from Huanghua City, Hebei province seashore pH 7.85, and organic 2.21%, full nitrogen 0.258%, available phosphorus 15.26mgkg
-1, full potassium 28.96gkg
-1, salt content 1.58%.
The preparation of 1.2 nanometer compost
Garbage compost 105 DEG C is dried to constant weight, by Qinhuangdao Tai Ji ring nanometer products Co., Ltd, utilizes the high-energy ball-milling process improved, by tank body multi-D swing motion fast, garbage compost processing grinding is made nanometer compost, for subsequent use.By granularmetric analysis under Electronic Speculum, nanometer compost particle size distribution is even, and average grain diameter is at about 30nm (Fig. 1).
The preparation of 1.3 Salt-tolerant microbial agent
1.3.1 the enrichment of bacterial classification
Compost sample is taken 10g and be placed in aseptic conical flask, after adding 100mL sterile water shaken well, get 10mL suspension in the conical flask filling 100mL enriched medium, at 28 DEG C, shaken cultivation 3d under 220r/min, is mixed microorganism flora.
1.3.2 the strengthening of salt-durable microbe
What the strengthening of mixed microorganism flora salt tolerant adopted is the method progressively increasing salinity, to alleviate the moment impact that causes hybrid bacterial strain of high salt concentration and murder by poisoning.Mixed bacterial without domestication in NaCl concentration be generally under the condition of 0.5% growth better, and the target NaCl concentration this time studied is 3%, namely in this research, the concentration of NaCl is respectively 0.5%, 1%, 1.5%, 2%, 2.5% and 3%.In domestication process, depending on OD
600increase and progressively improve salinity, the concentration of NaCl is increased to 3% with the gradient of 0.5%, often increases the concentration of a NaCl, after thalline increment is stable, could will continues the concentration of increase NaCl, tame out salt tolerant mixed microorganism flora step by step.
1.3.3 separation and the purifying of salt-durable microbe is strengthened
1.3.3.1 dilution spread flat band method
1) plate is down flat: by beef extract-peptone agar medium, Gao Shi No. I agar medium, Ma Dingshi (PDA) agar medium high-temperature sterilization, when being cooled to 55-60 DEG C, in martin agar medium, add Streptomycin Solution (final mass concentration is 0.3%), being mixed evenly is down flat plate respectively afterwards.
2) mixed microorganism dilution is prepared: add in the Boiling tube filling 9ml sterile water with the mixed microorganism bacteria suspension after liquid-transfering gun absorption 1ml strengthening and fully mix, this is 10
-1dilution, makes 10 by that analogy
-2, 10
-3, 10
-4, 10
-5with 10
-6the dilution of several concentration.
3) be coated with: the dilution bacteria suspension drawing 0.2ml variable concentrations with liquid-transfering gun respectively accurately puts into corresponding culture medium flat plate central authorities, and the process of each variable concentrations gradient repeats 3 times.Be coated with lightly evenly in media surface with sterile glass rod.
4) cultivate: beef extract-peptone flat-plate inverted is placed in 37 DEG C of incubators and cultivates, and the flat-plate inverted containing Gao Shi No. I medium and Ma Dingshi medium (PDA) is placed in 28 DEG C of incubators and cultivates 3-5 d.
1.3.3.2 plate streaking partition method
Choose bacterium colony: by the single bacterium colony that grows after cultivating respectively a little lawn of picking on new above-mentioned 3 kinds of medium, carry out line purifying.Until longer on medium is purebred, as impure, still needs to repeat this step, finally obtain 3 bacterial classifications.
1.3.4 the qualification of enhancement microbiological
The DNA of dominant bacteria is extracted according to the operation manual of kit.The PCR system of predominant bacteria: 10 × Buffer(with MgCl
2) 2 μ L, dNTP(10mmol/L) 0.4 μ L, 341f(10 μm of ol/L) 1 μ L, 534r(10 μm of ol/L) 1 μ L, Taq enzyme (5u/ μ L) 0.4 μ L, template DNA 1 μ L, add ultra-pure water and be settled to final volume 20 μ L.PCR reaction condition: 94 DEG C of 5min denaturations, 94 DEG C of sex change 1min, 55 DEG C of renaturation 45s, 72 DEG C extend 45s, 30 circulations, and 72 DEG C extend 10min.Primer is 341f (5 '-CGC CCG CCG CGC GCG GCG GGC GGG GCG GGG GCA CGG GGG GCC TAC GGG AGG CAG CAG-3 ') and 534r(5 '-ATT ACC GCG GCT GCT GG-3 ').The PCR reaction system of dominant fungi: 10 × Buffer (without MgCl
2) 2 μ L, MgCl
2(25mmol/L) 1.6 μ L, dNTP(10mmol/L) 0.4 μ L, Geo11(10 μm of ol/L) 0.4 μ L, GeoA2(10 μm of ol/L) 0.4 μ L, Taq enzyme (5u/ μ L) 0.2 μ L, template DNA 1 μ L, add ultra-pure water and be settled to final volume 20 μ L.PCR reaction condition: 94 DEG C of 4min denaturations, 94 DEG C of sex change 1min, 54 DEG C of renaturation 1min, 72 DEG C extend 2min, 30 circulations, and 72 DEG C extend 7min.Primer is GeoA2 (5 '-CCA GTA GTC ATA TGC TTG TCT C-3 ') and Geo11 (5 '-ACC TTG TTA CTT TTA CTT CC-3 ').The PCR primer obtained is delivered to Hua Da gene sequencing portion, Beijing, according to the result of order-checking, in BLAST system, find out corresponding bacterial classification.
Pass through its colony morphological observation above-mentioned 3 bacterial classifications, binding molecule qualification result (PCR sequence similarity) compares, and qualification result is respectively: bacillus subtilis, lysine bacillus and penicillium chrysogenum:
1.3.5 complex micro organism fungicide preparation
On corresponding liquid nutrient medium, cultivation is expanded by screening the strengthening salt-durable microbe obtained in previous experiments.Bacillus subtilis and lysine bacillus are at 30 DEG C, and 180r/min cultivates 1d, and penicillium chrysogenum is at 28 DEG C, and 220r/min cultivates.Cultivate after bacillus subtilis, lysine bacillus becomes complex micro organism fungicide with penicillium chrysogenum by the proportions of 1:1:1, dilute 100 times for subsequent use.
1.4 experimental designs and turf established by sod
Campus soil and saline-alkali soil to be sieved removal of impurities, make slight (0.3%), moderate (0.6%) and severe (0.9%) saline-alkali soil by a certain percentage, by for subsequent use for saline-alkali soil sterilizing 30 min at 121 DEG C mixed.Each coercing establishes 3 process, and three to coerce set process identical, is respectively:
(1) with 100% soil for contrast matrix add 15 ml blank cultures (CK)
(2) soil and nanometer compost mix in the ratio of 59:1 and add 15ml blank cultures (M1)
(3) soil and nanometer compost mix in the ratio of 59:1 and add 15ml strengthening Salt-tolerant microbial agent (M2)
With the soil mixed for matrix carries out lawn plant experiment in cultivation, during experiment, substrates quantity is 9400gm
-2, compounded turf matrix thickness is 15 mm, and Festuca Arundinacea grass seeds seeding quantity is 160gm
-2.Evenly sow after seed being soaked 24h in matrix top layer, first carry out dark processing, fully water and ensure to sprout, cultivate after sprouting, intensity of illumination is 450-700 μm of ol.m
-2.s
-1, period regularly waters a water sooner or later, often exchanges the position of cultivating turf consistent to ensure illumination.Indoor relative moisture is 40-55%, and temperature is 20-25 DEG C, carries out the mensuration of each index of plant after 50d is carried out in experiment.
The mensuration of 1.5 indexs
1.5.1 the mensuration of biomass
Finally neat for lawn plant matrix cradled, measure fresh weight on the ground, root is clean with distilled water flushing, and complete under being placed in baking oven 105 DEG C of conditions 30 min, dries to constant weight for 80 DEG C, measures on the ground and underground dry weight.
1.5.2 the mensuration of chlorophyll content
The mensuration of chlorophyll content: get 0.1 g blade, is cut into 1-2 mm fragment, is soaked in acetone: in ethanol (V:V=1:1) solution 24 hours, and soak is liquid to be measured.Under wavelength 633 nm and 645 nm, light absorption value is measured with spectrophotometer, and according to formulae discovery chlorophyll content:
1.6 data processing
Microsoft Excel and SPSS 17.0 software is adopted to process.
2 development results analyses
2.1 enhancement microbiological associating nanometer compost are on the impact of lawn plant biomass under salt stress
Salt stress obviously can suppress the accumulation of Festuca Arundinacea biomass, and along with the increase of salt stress degree, Festuca Arundinacea above and below ground biomass accumulation all obviously reduces (table 2).Singly execute nanometer compost and nanometer compost and enhancement microbiological co-administered, all can significantly improve the accumulation of Festuca Arundinacea biomass, strengthen the resistance to salt stress ability of plant.Wherein with the two co-administered effect significantly, under 0.3% and 0.6% salt stress, fresh weight adds 43.8% and 76.0% than contrast respectively on the ground, and when severe salt stress, fresh weight is 3 times of contrast on the ground, and this also has certain relation with sprouting.The two is co-administered also has obvious facilitation to root growth, and slight, moderate, Severe drought are coerced down, and underground dry weight is 1.5 times, 1.9 times and 3.7 times of contrast respectively, applies nanometer compost effect and takes second place.Experimental result shows, enhancement microbiological associating nanometer compost significantly can promote plant root growth, contributes to the raising of Festuca Arundinacea salt resistance.
Table 2 enhancement microbiological microbial inoculum associating nanometer compost is on impact (the g m of Festuca Arundinacea biomass under salt stress
-2)
2.4 enhancement microbiological associating nanometer compost are on the impact of Tall Fescue Leaves chlorophyll contents under salt stress
As shown in Table 3, along with the increase of salt stress degree, Festuca Arundinacea chlorophyll content in leaf blades reduces gradually, and under severe water stress condition, leaves of plants lamellar body inner chlorophyll content is minimum.Under same salt stress degree, nanometer compost and enhancement microbiological microbial inoculum are combined to apply and obviously can be promoted the chlorophyllous increase of Festuca Arundinacea, have obvious facilitation to Festuca Arundinacea photosynthetic rate.Higher chlorophyll content is all shown under three kinds of salt stress levels, slightly, under moderate and severe water stress, Chlorophyll content exceeds 12.34%, 24.08% and 30.59% than contrast respectively, Chlorophyll-a Content exceeds 18.2%, 25.0% and 29.7% than contrast respectively, and chlorophyll b also has increase in various degree.
Table 3 enhancement microbiological microbial inoculum associating nanometer compost is on the impact (mg.g of Tall Fescue Leaves chlorophyll contents under salt stress
-1fW)
3 conclusions
Experimental result shows, enhancement microbiological associating nanometer compost can significantly improve lawn plant salt resistance, and still upgrowth situation is good when salt content is 0.9% to ensure lawn plant.Slightly and under moderate salt stress also there is positive effect, after applying enhancement microbiological associating nanometer compost, fresh weight adds 12.1% and 76% than contrast respectively on the ground, and during severe salt stress, fresh weight is 3 times more than of contrast on the ground, and underground dry weight is 1.5 times, 1.8 times and 3 times of contrast respectively.
Accompanying drawing illustrates:
Granularmetric analysis under Fig. 1 nanometer compost Electronic Speculum.
Embodiment
In order to explain enforcement of the present invention more fully, provide following preparation method's embodiment.These embodiments are only explain instead of limit the scope of the invention.Special instruction is needed to be: the present invention screens in the complex micro organism fungicide obtained, and all there is sale in bacillus subtilis, lysine bacillus and penicillium chrysogenum market, also method of the present invention can be adopted to be separated from consumer garbage compost and obtain complex micro organism fungicide, the biochemical characteristic of its flora obtained is identical with commercially available therefore not in preservation.
Embodiment 1
Adopt salt strengthening active nano garbage compost to strengthen the method for Salinity Tolerance of Turfgrass, it is characterized in that being undertaken by following step: the preparation of (1) nanometer compost:
Consumer garbage compost 105 DEG C is dried to constant weight, garbage compost processing grinding is made 30nm nanometer compost, for subsequent use;
(2) preparation of Salt-tolerant microbial agent:
Compost sample is taken 10g and be placed in aseptic conical flask, after adding 100mL sterile water shaken well, get 10mL suspension in the conical flask filling 100mL enriched medium, at 28 DEG C, shaken cultivation 3d under 220r/min, is mixed microorganism flora; Containing bacillus subtilis, lysine bacillus and penicillium chrysogenum in described mixed microorganism flora;
(3) what the strengthening of mixed microorganism flora salt tolerant adopted is the method progressively increasing salinity, to alleviate the moment impact that causes hybrid bacterial strain of high salt concentration and murder by poisoning, the concentration of NaCl is increased to 3% with the gradient of 0.5%, often increase the concentration of a NaCl, after thalline increment is stable, the concentration increasing NaCl could be continued, tame out salt-durable microbe flora step by step;
(4) preparation of compound Salt-tolerant microbial agent:
The proportions of bacillus subtilis, lysine bacillus and penicillium chrysogenum 1:1:1 by volume becomes compound Salt-tolerant microbial agent, dilute 100 times for subsequent use;
(5) turf established by sod:
Soil and saline-alkali soil to be sieved removal of impurities, make 0.3% slight (W/W), 0.6% moderate (W/W) and 0.9% severe (W/W) saline-alkali soil by a certain percentage, by for subsequent use for saline-alkali soil sterilizing 30 min at 121 DEG C mixed, each coercing establishes 3 process, three to coerce set process identical, is respectively:
(1) with 100% soil for contrast matrix add 15 ml blank cultures;
(2) soil and nanometer compost mix in the ratio of 59:1 and add 15ml blank cultures;
(3) soil and nanometer compost mix in the ratio of 59:1 and add 15ml strengthening Salt-tolerant microbial agent;
With the soil mixed for matrix carries out lawn plant experiment in cultivation, during experiment, substrates quantity is 9400gm
-2, compounded turf matrix thickness is 15 mm, and Festuca Arundinacea grass seeds seeding quantity is 160gm
-2, son is evenly sowed in matrix top layer after soaking 24h, first carries out dark processing, and fully water and ensure to sprout, cultivate after sprouting, intensity of illumination is 450 μm of ol.m
-2.s
-1, period regularly waters a water sooner or later, and often exchange the position of cultivation turf to ensure that illumination is consistent, indoor relative moisture is 40%, and temperature is 20 DEG C, carries out the mensuration of each index of plant after 50d is carried out in experiment.The described method progressively increasing salinity refers to: the concentration of NaCl is respectively 0.5%, 1%, 1.5%, 2%, 2.5% and 3%, in domestication process, depending on OD
600increase and progressively improve salinity, the concentration of NaCl is increased to 3% with the gradient of 0.5%.Test material comprises:
(1) Festuca Arundinacea: choose full seed, perennial Festuca Arundinacea of uniform size (
festuca arundinaceal.) seed is test material;
(2) consumer garbage compost: from Tianjin little Dian composting plant, basic physical and chemical is: pH 7.62, saturation moisture content 0.76 ml/g, unit weight 0.85 g/ml, full nitrogen 5.18%, full potassium 50.83 g/kg, available phosphorus 77.92 mg/kg, organic 12.12%, garbage compost is removed the foreign material such as wood, plastics, metal wherein, air-dry rear for subsequent use;
(3) soil: impurities removing, cross 2mm sieve, be positioned over ventilating opening place 2d, air-dry under natural conditions, its physicochemical property is: pH7.44, saturation moisture content 0.58mLg
-1, organic 4.68%, full nitrogen 0.21%, salt content 0.29%;
(4) saline-alkali soil: take from Huanghua City, Hebei province seashore pH 7.85, organic 2.21%, full nitrogen 0.258%, available phosphorus 15.26mgkg
-1, full potassium 28.96gkg
-1, salt content 1.58%.
Embodiment 2
Adopt salt strengthening active nano garbage compost to strengthen the method for Salinity Tolerance of Turfgrass, it is characterized in that being undertaken by following step: the preparation of (1) nanometer compost:
Consumer garbage compost 105 DEG C is dried to constant weight, garbage compost processing grinding is made 35nm nanometer compost, for subsequent use; (2) preparation of Salt-tolerant microbial agent:
Compost sample is taken 10g and be placed in aseptic conical flask, after adding 100mL sterile water shaken well, get 10mL suspension in the conical flask filling 100mL enriched medium, at 28 DEG C, shaken cultivation 3d under 220r/min, is mixed microorganism flora; Containing bacillus subtilis, lysine bacillus and penicillium chrysogenum in described mixed microorganism flora;
(3) what the strengthening of mixed microorganism flora salt tolerant adopted is the method progressively increasing salinity, to alleviate the moment impact that causes hybrid bacterial strain of high salt concentration and murder by poisoning, the concentration of NaCl is increased to 3% with the gradient of 0.5%, often increase the concentration of a NaCl, after thalline increment is stable, the concentration increasing NaCl could be continued, tame out salt-durable microbe flora step by step;
(4) preparation of compound Salt-tolerant microbial agent:
The proportions of bacillus subtilis, lysine bacillus and penicillium chrysogenum 1:1:1 by volume becomes compound Salt-tolerant microbial agent, dilute 100 times for subsequent use;
(5) turf established by sod:
Soil and saline-alkali soil to be sieved removal of impurities, make 0.3% slight (W/W), 0.6% moderate (W/W) and 0.9% severe (W/W) saline-alkali soil by a certain percentage, by for subsequent use for saline-alkali soil sterilizing 30 min at 121 DEG C mixed, each coercing establishes 3 process, three to coerce set process identical, is respectively:
(1) with 100% soil for contrast matrix add 15 ml blank cultures;
(2) soil and nanometer compost mix in the ratio of 59:1 and add 15ml blank cultures;
(3) soil and nanometer compost mix in the ratio of 59:1 and add 15ml strengthening Salt-tolerant microbial agent;
With the soil mixed for matrix carries out lawn plant experiment in cultivation, during experiment, substrates quantity is 9400gm
-2, compounded turf matrix thickness is 15 mm, and Festuca Arundinacea grass seeds seeding quantity is 160gm
-2, son is evenly sowed in matrix top layer after soaking 24h, first carries out dark processing, and fully water and ensure to sprout, cultivate after sprouting, intensity of illumination is 700 μm of ol.m
-2.s
-1, period regularly waters a water sooner or later, and often exchange the position of cultivation turf to ensure that illumination is consistent, indoor relative moisture is 40-55%, and temperature is 25 DEG C, carries out the mensuration of each index of plant after 50d is carried out in experiment.The described method progressively increasing salinity refers to: the concentration of NaCl is respectively 0.5%, 1%, 1.5%, 2%, 2.5% and 3%, in domestication process, depending on OD
600increase and progressively improve salinity, the concentration of NaCl is increased to 3% with the gradient of 0.5%.Test material comprises:
(1) Festuca Arundinacea: choose full seed, perennial Festuca Arundinacea of uniform size (
festuca arundinaceal.) seed is test material;
(2) consumer garbage compost: from Tianjin little Dian composting plant, basic physical and chemical is: pH 7.62, saturation moisture content 0.76 ml/g, unit weight 0.85 g/ml, full nitrogen 5.18%, full potassium 50.83 g/kg, available phosphorus 77.92 mg/kg, organic 12.12%, garbage compost is removed the foreign material such as wood, plastics, metal wherein, air-dry rear for subsequent use;
(3) soil: impurities removing, cross 2mm sieve, be positioned over ventilating opening place 3d, air-dry under natural conditions, its physicochemical property is: pH7.44, saturation moisture content 0.58mLg
-1, organic 4.68%, full nitrogen 0.21%, salt content 0.29%;
(4) saline-alkali soil: take from Huanghua City, Hebei province seashore pH 7.85, organic 2.21%, full nitrogen 0.258%, available phosphorus 15.26mgkg
-1, full potassium 28.96gkg
-1, salt content 1.58%.
Claims (5)
1. adopt salt to strengthen a method for active nano garbage compost enhancing Salinity Tolerance of Turfgrass, it is characterized in that being undertaken by following step:
(1) soil and saline-alkali soil sieve removal of impurities, make slight 0.3%(w/w), moderate 0.6%(w/w) and severe 0.9%(w/w) saline-alkali soil, by for subsequent use for saline-alkali soil sterilizing 30 min at 121 DEG C mixed;
(2) soil of step (1) and 30-35nm nanometer compost mix in the ratio of 59:1 and add 15ml compound Salt-tolerant microbial agent; Described compound Salt-tolerant microbial agent refers to: bacillus subtilis: lysine bacillus: the proportions of penicillium chrysogenum 1:1:1 by volume;
(3) be that matrix carries out lawn plant experiment in cultivation with the step mixed (2) soil, during experiment, substrates quantity is 9400gm
-2, compounded turf matrix thickness is 15 mm, and Festuca Arundinacea grass seeds seeding quantity is 160gm
-2, evenly sow after seed being soaked 24h in matrix top layer, first carry out dark processing, fully water and ensure to sprout, cultivate after sprouting, intensity of illumination is 450-700 μm of ol.m
-2.s
-1, period regularly waters a water sooner or later, and often exchange the position of cultivation turf to ensure that illumination is consistent, indoor relative moisture is 40-55%, and temperature is 20-25 DEG C, carries out the mensuration of each index of plant after 50d is carried out in experiment.
2. described in claim 1, adopt salt to strengthen the method for active nano garbage compost enhancing Salinity Tolerance of Turfgrass, wherein:
(1) preparation of nanometer compost: consumer garbage compost 105 DEG C is dried to constant weight, garbage compost processing grinding is made 30-35nm nanometer compost, for subsequent use;
(2) turf established by sod:
Soil and saline-alkali soil to be sieved removal of impurities,=make 0.3% slight (W/W), 0.6% moderate (W/W) and 0.9% severe (W/W) saline-alkali soil, by for subsequent use for saline-alkali soil sterilizing 30 min at 121 DEG C mixed, each coercing establishes 3 process, three to coerce set process identical, is respectively:
(1) with 100% soil for contrast matrix add 15 ml blank cultures;
(2) soil and nanometer compost mix in the ratio of 59:1 and add 15ml blank cultures;
(3) soil and nanometer compost mix in the ratio of 59:1 and add 15ml strengthening Salt-tolerant microbial agent.
3. described in claim 1, adopt salt to strengthen the method for active nano garbage compost enhancing Salinity Tolerance of Turfgrass, the wherein said method progressively increasing salinity refers to: the concentration of NaCl is respectively 0.5%, 1%, 1.5%, 2%, 2.5% and 3%, in domestication process, increase depending on OD600 and progressively improve salinity, the concentration of NaCl is increased to 3% with the gradient of 0.5%.
4. the method adopting salt to strengthen active nano garbage compost enhancing Salinity Tolerance of Turfgrass described in claim 1 is improving the application in plant root growth.
5. adopt salt to strengthen active nano garbage compost described in claim 1 and strengthen the method for Salinity Tolerance of Turfgrass in the chlorophyllous increase of promotion Festuca Arundinacea, improve the application of Festuca Arundinacea photosynthetic rate aspect.
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