CN104844345A - Method for improving drought resistance of field turf by adopting drought tolerance-reinforced active nanometer waste compost - Google Patents
Method for improving drought resistance of field turf by adopting drought tolerance-reinforced active nanometer waste compost Download PDFInfo
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Abstract
The invention discloses a method for improving the drought resistance of field turf by adopting drought tolerance-reinforced active nanometer waste compost. The method is characterized in that appropriate beneficial microbe strains are separated and extracted from domestic waste compost, drought tolerance reinforcement is carried out to obtain an efficient microbial agent, the efficient microbial agent and nanometer compost are mixed and then are applied to a turf matrix, the drought tolerance of turf plants under drought stress is researched, and bases are provided for application of the nanometer compost combined microbial agent in the drought resistance of turf grass. Experiment results show that nanometer compost combined microbes are helpful for growth of Festuca arundinacea under the drought stress, improve the activity of leaf protection enzyme, enhance the adaptation ability of the plants to the drought stress, and realize the water saving rates under moderate stress and serious stress of 43.10% and 52.39% respectively.
Description
Technical field
The invention belongs to environmental protection technical field, relate to a kind of method adopting strengthening drought-enduring microbiobacterial agent associating nanometer compost to improve turf drought resistance.
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, Materials 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 nanotechnology, and nanostructure and nano material is considered as the key of nanotechnology and is preferentially supported.2012, Chinese industrial and information-based portion are according to the spirit of " country's " 12 " emerging strategic industries development program ", issue " new material industry " 12 " development program ", 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 cell levels and molecular level, as the impact etc. on hemocyte, lung cells, immunocyte, DNA damage, genetic expression.Domesticly there have been developed the impact of nano material on plant, nano fertilizer is proposed by soil and fertilizer 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 microcapsulary and make, is divided into nano material coating, cementing sustained-release and controlled release fertilizer and nanostructure fertilizer.As everyone knows, the groundwater pollution 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 bondss, so have nonsaturation, be easy to combine with other atoms and settle out, thus show very high chemically reactive.Can significantly improve the utilization ratio 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 materials.On the other hand, be because its surface effects, nano fertilizer surface energy and Surface binding energy are increased, and help it to be absorbed by root system of plant in edatope, 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 utilization ratio.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 fertilizer is also called bacterial manure, bio-feritlizer, 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 fertilizer 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 microorganisms 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.Microbiobacterial 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 microbial preparation energy Promoting plant growth and nutritive element.Have investigator 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 kind of plant growth-promoting fungies filtered out are applied to Sunflower Receptacle to be sent out, result proves, fungi can obviously promote its primary growth, and effectively can suppress the generation of oidium.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 microorganism there is no bibliographical information.
Turf establishment system integrates ecological 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 all respects 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, sterilization and disinfection, and lawn absorbs CO by photosynthesis
2release O
2keep CO in air
2and O
2balance, lawn can also be diluted, decompose, absorb and the toxic substance such as toxic gas, carcinogenic substance and some heavy metal gas in fixing air, is turned bane into boon by photosynthesis.Secondly, lawn can conserve water and soil, maintain the eubiosis.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 atmospheric moisture.
In recent years, environmental problem is day by day serious, and green grassland has very important ecology and the value of environmental protection in protection environment for human survival and human physical and mental health.But turfgrass survives in nature, geographic range is extensive, and growth and breeding often suffers the impact of poor environment, as having distribution in arid, saline and alkaline and cold wet environment.Wherein arid is one of main environment stress factors of restriction Turfgrass Growth.Arid causes water deficient in plant materials, suppresses plant-growth, more seriously causes plant mechanical injuries thus causes plant dead.China is vast in territory, and weather is various, and wherein 47 % areas are arid and semi-arid area, and shortage of water resources has become China's most area even global problem, significantly limit the growth planting of Lawn, has increased the weight of the difficulty of turf production management.Moisture ensures the important factor of Turfgrass Growth, and lawn water deficient can make lawn accelerated deterioration, shortens lawn working life.In addition, cultivation lawn, city drought-resistance ability is relatively poor compared with natural lawn, and its lawn root system is shallow, intake water scarce capacity, needs to carry out man-made irrigation.In northern China city, the year irrigation quantity on every square metre of lawn is approximately 0.6-1.0 m
3.So in today that the nervous environment of water resources constantly worsens, the ultimate challenge that turf establishment faces is allotment and the utilization of water resources.Thus turf grass drought resistance research is to saving irrigation water, promotes the planting of drought environment lawn plant, alleviates water resources crisis and just seems most important.
For above problem, current research mainly concentrate on choose strong stress resistance lawn plant, use concentrated fertilizer, use the measure such as water-holding agent 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 edatope, 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, there is no bibliographical information.
Consumer garbage compost is one and highly oozes system, and microorganism wherein has certain adaptive faculty to adverse circumstance, will wherein beneficial microorganism 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 microorganism in turf establishment thereof, garbage compost is worked into about 30nm through nanofabrication technique by this technology, 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, with nanometer compost be united and applied in drought stress coerce under turf establishment, 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.And experimental result is applied to field experiment, further checking has been done, for practical application provides foundation to the effect of enhancement microbiological associating nanometer compost in the environment stress of field.
By measuring strengthening drought-enduring microbiobacterial agent associating nanometer compost, in field to Index Influences such as lawn plant plant height, green degree, cover degree, biomass and green phases, to the practical application effect of research enhancement microbiological microbial inoculum associating nanometer compost in land for growing field crops, for its practical application provides technical support.
Summary of the invention
The present invention goes out suitable beneficial microbe strain by separation and Extraction from consumer garbage compost, more efficient microbiobacterial agent is obtained by drought-enduring strengthening, itself and nanometer compost is co-administered in lawn matrix, study its drought tolerance to lawn plant under drought stress, for the application of nanometer compost associating microbiobacterial agent in turf grass drought resistance provides foundation.
The invention discloses following technology contents for achieving the above object:
Adopt drought-enduring strengthening active nano garbage compost to improve a method for field sod production drought resistance, 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 drought-enduring microbiobacterial agent
Compost sample is taken 10g and be placed in aseptic Erlenmeyer flask, after adding 100mL sterilized water shaken well, get 10mL suspension in the Erlenmeyer flask filling 100mL enrichment medium, at 28 DEG C, shaking culture 3d under 220r/min, is mixing microorganisms flora; What the drought-enduring strengthening of mixing microorganisms flora adopted is the method progressively increasing PEG6000 concentration, in strengthening process, depending on OD
600increase and progressively improve PEG6000 concentration, the concentration of PEG6000 increases with the gradient of 5%, and ultimate aim is 25%;
Compost sample is taken 10g and be placed in aseptic Erlenmeyer flask, after adding 100mL sterilized water shaken well, get 10mL suspension in the Erlenmeyer flask filling 100mL enrichment medium, at 28 DEG C, shaking culture 3d under 220r/min, is mixing microorganisms flora; The low temperature resistant strengthening of mixing microorganisms is by different concns gradient (10
-1, 10
-2, 10
-3, 10
-4, 10
-5with 10
-6) be coated on (i.e. beef-protein medium, Gao Shi No. I substratum and Ma Dingshi substratum) on corresponding substratum be placed in artificial climate incubator cultivate 2 weeks, temperature is 10 DEG C;
What the drought-enduring strengthening of mixing microorganisms flora adopted is the method progressively increasing PEG6000 concentration.Mixed bacterial without strengthening generally grows better, in strengthening process, depending on OD under without the condition of PEG6000
600increase and progressively improve PEG6000 concentration, the concentration of PEG6000 increases with the gradient of 5%, and ultimate aim is 25%; Often increase the concentration of a PEG6000, after thalline increment is stable, will could continues the concentration increasing PEG6000, strengthen step by step and drought-enduring mixing microorganisms flora, contain;
(3) preparation of the drought-enduring microbiobacterial agent of compound
Three kinds of drought-enduring microorganisms of the strengthening enlarged culturing in corresponding liquid nutrient medium obtained will be screened, bacillus cereus and Methionin genus bacillus are at 30 DEG C, 180r/min cultivates, rhodotorula glutinis is at 28 DEG C, 220r/min cultivates, cultivate after bacillus cereus, Methionin genus bacillus and rhodotorula glutinis by volume 1:1:1 be mixed with composite microbial bacteria liquid, dilute 100 times and 4 times for subsequent use;
(4) experiment establishes 4 process, each process repetition 3 times, and plot area is 6m
2(3m × 2m), each process adopts the group arrangement of random district, and a stress level is established in experiment, and experimental session hillslope processes maintains severe water stress (40% ~ 20% FC), and experimental design is as follows:
1) 1L blank cultures is added
2) add 8g nanometer compost and add 1L blank cultures
3) add 8g nanometer compost and add the complex micro organism fungicide that 1L dilutes 100 times
4) add 8g nanometer compost and add the complex micro organism fungicide that 1L dilutes 4 times
(5) after process, in 4-11 in 2014 month-carry out plant height mensuration, take the mode of random sampling to measure at every turn.
Wherein: the concentration of PEG6000 is respectively 5%, 10%, 15%, 20% and 25%.Often increase the concentration of a PEG6000, after thalline increment is stable, will could continues the concentration increasing PEG6000, strengthen step by step and drought-enduring mixing microorganisms flora.Material therefor:
Choose full seed, perennial Festuca Arundinacea of uniform size (
festuca arundinacea L.) seed is test materials;
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 impurities removing, cross 2mm sieve, be positioned over ventilation opening place 2-3d, air-dry under natural condition, its physico-chemical property is: pH 7.44, saturation moisture content 0.58mL/g, organic 4.68%, full nitrogen 0.21%, available phosphorus 22.03 mg/kg, full potassium 45.61g/kg.
The present invention further discloses the method adopting drought-enduring strengthening active nano garbage compost to improve field sod production drought resistance and promote the application in the green degree anaphase effect of turf.Particularly extend the turf green phase, in advance turn green the time, improve application in regrowth rate.
The more detailed preparation method of the present invention is as follows:
1 development materials and methods
1.1 experiment material
Experiment field is located in the institute of grass flowers and plants technique center, the Tianjin North Star red port, is located in N 39.23′，E 117。21 ', height above sea level 5m, average temperature of the whole year 12.1 DEG C, the lowest temperature-10 DEG C, the highest temperature 31.2 DEG C.Experimental plot basic physical and chemical is: pH 7.7, saltiness 0.3%, full nitrogen 0.32%, full K43 .27 g/kg, available phosphorus 22.41mg/kg, organic 4.83%, and for examination lawn by English grass and English ryegrass 9:1 ratio mixed seeding in mass ratio, planting 6 years.
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.
1.2 trial plot moisture controlled and management
2014 annual precipitation 293.8mm, focus mostly in 7 ~ September, and process initial moisture in experiment field is 15-20%, and substantially anhydrous point of irrigation after process is annual by natural rainfall, but Winter-Spring is almost without precipitation, maintains severe water stress state.
The preparation of 1.3 nanometer compost
Garbage compost 105 DEG C is dried to constant weight, by Qinhuangdao Tai Ji ring nanometer products company limited, 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 particle size analysis under Electronic Speculum, nanometer compost particle size distribution is even, and median size is at about 30nm.
The preparation of 1.4 drought-enduring microbiobacterial agents
1.4.1 the enrichment of bacterial classification
Compost sample is taken 10g and be placed in aseptic Erlenmeyer flask, after adding 100mL sterilized water shaken well, get 10mL suspension in the Erlenmeyer flask filling 100mL enrichment medium, at 28 DEG C, shaking culture 3d under 220r/min, is mixing microorganisms flora.
1.4.2 the strengthening of drought-enduring microorganism
What the drought-enduring strengthening of mixing microorganisms flora adopted is the method progressively increasing PEG6000 concentration.Mixed bacterial without strengthening generally grows better, in strengthening process, depending on OD under without the condition of PEG6000
600increase and progressively improve PEG6000 concentration, the concentration of PEG6000 increases with the gradient of 5%, and ultimate aim is 25%; Namely the concentration of PEG6000 is respectively 5%, 10%, 15%, 20% and 25% in this experiment.Often increase the concentration of a PEG6000, after thalline increment is stable, will could continues the concentration increasing PEG6000, strengthen step by step and drought-enduring mixing microorganisms flora.
1.4.3 separation and the purifying of drought-enduring microorganism is strengthened
1.4.3.1 dilution spread flat band method
1) plate is down flat: by beef extract-peptone nutrient agar, Gao Shi No. I nutrient agar, Ma Dingshi (PDA) nutrient agar high-temperature sterilization, when being cooled to 55-60 DEG C, in martin agar substratum, add Streptomycin Solution (final quality concentration is 0.3%), being mixed evenly is down flat plate respectively afterwards.
2) mixing microorganisms diluent is prepared: add in the Boiling tube filling 9ml sterilized water with the mixing microorganisms bacteria suspension after liquid-transfering gun absorption 1ml strengthening and fully mix, this is 10
-1diluent, makes 10 by that analogy
-2, 10
-3, 10
-4, 10
-5with 10
-6the diluent of several concentration.
3) be coated with: the dilution bacteria suspension drawing 0.2ml different concns with liquid-transfering gun respectively accurately puts into corresponding culture medium flat plate central authorities, and the process of each different concns 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 substratum and Ma Dingshi substratum (PDA) is placed in 28 DEG C of incubators and cultivates 3-5 d.
1.4.3.2 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 substratum, carry out line purifying.Until longer on substratum is purebred, as impure, still needs to repeat this step, finally obtain 3 bacterial classifications.
1.4.4 strengthen the qualification of drought-enduring microorganism
The DNA of dominant bacteria is extracted according to the operational manual of test 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 ultrapure water and be settled to final volume 20 μ L.PCR reaction conditions: 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 ultrapure water and be settled to final volume 20 μ L.PCR reaction conditions: 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 isolated bacterial classification to its colony morphological observation, binding molecule qualification result (PCR sequence similarity) compares, and identifies and is respectively: bacillus cereus, rhodotorula glutinis and Methionin genus bacillus.
Drought-enduring microorganism identification result strengthened by table 1
By three kinds of drought-enduring microorganisms of strengthening enlarged culturing in corresponding liquid nutrient medium.Bacillus cereus and Methionin genus bacillus are at 30 DEG C, and 180r/min cultivates, and rhodotorula glutinis is at 28 DEG C, and 220r/min cultivates.Cultivate after bacillus cereus, Methionin genus bacillus and rhodotorula glutinis are mixed with composite microbial bacteria liquid by the volume ratio of 1:1:1, dilute 100 times and 4 times for subsequent use.
1.5 experimental design
Experiment establishes 4 process, each process repetition 3 times, and plot area is 6m
2(3m × 2m), each process adopts the group arrangement of random district, and a stress level is established in experiment, and experimental session hillslope processes maintains severe water stress (40% ~ 20% FC).Experimental design is as follows:
(1) 1L blank cultures (CK) is added
(2) add 8g nanometer compost and add 1L blank cultures (M1)
(3) add 8g nanometer compost and add the complex micro organism fungicide (M2) that 1L dilutes 100 times
(4) add 8g nanometer compost and add the complex micro organism fungicide (M3) that 1L dilutes 4 times.
1.6 testing index
1.6.1 the mensuration of plant height
After process, carry out plant height mensuration on April 20th, 2014, on April 30th, 2014, on May 16th, 2014, on May 26th, 2014, on November 3rd, 2014, take the mode of random sampling to measure, each process repetition 6 times at every turn.
1.6.2 the mensuration of biomass
Each process chooses 0.01m immediately
2the sample prescription of (0.1m × 0.1m), cradles neat for lawn plant matrix, and 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.
1.6.3 chlorophyllous mensuration
The portable Chlorophyll meter of SPDA-502 that chlorophyll adopts Japanese Konica Minolta to produce measures.Measure 2 times altogether, minute is respectively on November 3rd, 2014 and on December 18th, 2014.
1.6.4 the mensuration of green degree
After process, carry out green degree measure on April 20th, 2014, on April 30th, 2014, on May 16th, 2014, on May 26th, 2014, on November 3rd, 2014, adopt appearance method, nine points of systems evaluate turf quality, color and luster evaluation is done by 4 valuators are relatively independent in each community, averages as evaluation result.
1.6.5 the mensuration of leaf area index
Leaf area index is adopted as Si Aidi (Beijing) ecological science Instrument Ltd. and produces CI-203 Handheld laser leaf area instrument, and the same portion choosing relatively homogeneous turfgrass blade is positioned at and measures on November 3rd, 2014.
1.6.6 withered and yellow phase and regrowth rate
Observe the green phase length of turf, record the withered and yellow date, measure turn green time and regrowth rate spring in Second Year.Thus calculate the turf green phase, namely start to turn green to completely withered and yellow the experienced time.
1.6.7 the mensuration of cover degree
After process, carry out cover degree mensuration on May 26th, 2014, adopt appearance method, cover degree evaluation is done by 4 valuators are relatively independent in each community, averages as evaluation result.
1.7 data analyses adopt Microsoft Excel and SPSS17.0 software to process.
2 development results analyses
2.1 enhancement microbiological associating nanometer compost are on the impact of turfgrass plant height under field condition
Under different treatment, the dynamic plant height of Festuca Arundinacea is in table 2.As can be seen from the table, Severe drought is coerced down, and enhancement microbiological associating nanometer compost has obvious promoter action to lawn plant plant height.Wherein, with the best results of nanometer compost high concentration enhancement microbiological, during on May 16th, 2014, improve 86.1% compared with the control.
Table 2 enhancement microbiological associating nanometer compost is on the impact (cm) of lawn plant plant height under the drought stress of field
The impact of turfgrass biomass under 2.2 enhancement microbiological associating nanometer compost field drought stress conditions
As can be seen from Table 3, under drought stress, each process community lawn plant biomass is all apparently higher than check plot.Wherein with M3 nanometer compost high concentration enhancement microbiological microbial inoculum treatment group best results, Severe drought is coerced down, and M3 treatment group on the ground fresh weight, on the ground dry weight and underground dry weight is respectively 2.0 times, 2.2 times and 3.7 times more than of control group, and difference is extremely remarkable.
Table 3 enhancement microbiological associating nanometer compost is on the impact (g/m of lawn plant biomass under the drought stress of field
-2)
2.3 enhancement microbiologicals associating nanometer compost are on the impact of the grass green degree in lawn under autumn has set in phase field drought stress
As shown in Table 4, enhancement microbiological associating nanometer compost is comparatively remarkable to the lifting anaphase effect of the green degree of turf, and during on May 26th, 2014, the green degree in M3 treatment group lawn exceeds 25.5% than contrast.
Table semi-finals microbial association nanometer compost is on the impact of the green degree of lawn plant under the drought stress of field
2.4 enhancement microbiological associating nanometer compost are on the chlorophyllous impact of lawn plant under the drought stress of field
As shown in Table 5, three treatment group lawn plant chlorophyll contents are all significantly higher than control group, and wherein enhancement microbiological associating nanometer compost treatment group effect is better than single compost treatment group.Severe drought is coerced down, and M3 treatment group turfgrass chlorophyll a, chlorophyll b, Chlorophyll content, exceed 27.9%, 70.4% and 36.6% than contrast respectively, and between interpolation high density microbial inoculum and the process of lower concentration microbial inoculum, there is some difference.
Table 5 enhancement microbiological associating nanometer compost is on the chlorophyllous impact (mg.g of lawn plant under the drought stress of field
-1fW)
2.5 enhancement microbiological associating nanometer compost are on the impact of lawn plant leaf area index under the drought stress of field
Apply enhancement microbiological associating nanometer compost to the impact of turfgrass leaf area index under drought stress in table 6.Enhancement microbiological associating nanometer compost effectively can promote the growth of lawn plant blade under drought stress, and showing as blade length increases, and area strengthens, and wherein M3 treatment group blade length and leaf girth add 249.8% and 71.5% compared with the control respectively.
Table 6 enhancement microbiological associating nanometer compost is on the impact of lawn plant leaf area index under the drought stress of field
2.6 enhancement microbiological associating nanometer compost are on the impact of turfgrass regrowth rate under the drought stress of field
From table 7, obviously can find out that being applied with enhancement microbiological associating nanometer compost significantly can promote the prolongation of green period of lawn and the raising of regrowth rate, the green phase extends 15 days to M3 compared with the control, and regrowth rate improves 15%.
Table 7 enhancement microbiological associating nanometer compost is on the impact of field lawn plant green phase and regrowth rate
2.7 enhancement microbiological associating nanometer compost are on the impact of turfgrass cover degree under the drought stress of field
2014.5.26 day measures lawn, different treatment community coverage under drought stress, and result is as shown in table 8, singly executes nanometer compost and enhancement microbiological associating nanometer compost treatment all can improve lawn coverage, significant difference compared with the control.
Table Final 8 microbial association nanometer compost is on the impact of field lawn plant coverage under salt stress
3 development conclusions
In the application of this technology, microbial association nanometer compost effectively can improve the growth of turfgrass plant, the accumulation of biomass and chlorophyllous content improve, and this is conducive to improving, and lawn plant is photosynthetic to carry out and anti-environment stress ability.Wherein, add 4 times of diluting high-concentration microbiobacterial agent best results, fresh weight and underground dry weight compared with the control, are 1.7 times and 2.5 times more than of contrast respectively, can significantly improve the adaptive faculty of turfgrass to drought stress on the ground for they.The green phase prolongation of turf in addition, time advance of turning green, regrowth rate improve, cover degree increases, and these indexs improve the function value of turf greatly, have important practical application meaning.
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 bacillus cereus in the complex micro organism fungicide obtained, Methionin genus bacillus and rhodotorula glutinis market all there is sale, 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
A kind of method adopting drought-enduring strengthening active nano garbage compost to improve field sod production drought resistance:
(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 drought-enduring microbiobacterial agent
Compost sample is taken 10g and be placed in aseptic Erlenmeyer flask, after adding 100mL sterilized water shaken well, get 10mL suspension in the Erlenmeyer flask filling 100mL enrichment medium, at 28 DEG C, shaking culture 3d under 220r/min, is mixing microorganisms flora; What the drought-enduring strengthening of mixing microorganisms flora adopted is the method progressively increasing PEG6000 concentration, in strengthening process, depending on OD
600increase and progressively improve PEG6000 concentration, the concentration of PEG6000 increases with the gradient of 5%, and ultimate aim is 25%;
Compost sample is taken 10g and be placed in aseptic Erlenmeyer flask, after adding 100mL sterilized water shaken well, get 10mL suspension in the Erlenmeyer flask filling 100mL enrichment medium, at 28 DEG C, shaking culture 3d under 220r/min, is mixing microorganisms flora; The low temperature resistant strengthening of mixing microorganisms is by different concns gradient (10
-1, 10
-2, 10
-3, 10
-4, 10
-5with 10
-6) be coated on (i.e. beef-protein medium, Gao Shi No. I substratum and Ma Dingshi substratum) on corresponding substratum be placed in artificial climate incubator cultivate 2 weeks, temperature is 10 DEG C;
What the drought-enduring strengthening of mixing microorganisms flora adopted is the method progressively increasing PEG6000 concentration.Mixed bacterial without strengthening generally grows better, in strengthening process, depending on OD under without the condition of PEG6000
600increase and progressively improve PEG6000 concentration, the concentration of PEG6000 increases with the gradient of 5%, and ultimate aim is 25%; Often increase the concentration of a PEG6000, after thalline increment is stable, will could continues the concentration increasing PEG6000, strengthen step by step and drought-enduring mixing microorganisms flora, contain;
(3) preparation of the drought-enduring microbiobacterial agent of compound
Three kinds of drought-enduring microorganisms of the strengthening enlarged culturing in corresponding liquid nutrient medium obtained will be screened, bacillus cereus and Methionin genus bacillus are at 30 DEG C, 180r/min cultivates, rhodotorula glutinis is at 28 DEG C, 220r/min cultivates, cultivate after bacillus cereus, Methionin genus bacillus and rhodotorula glutinis by volume 1:1:1 be mixed with composite microbial bacteria liquid, dilute 100 times and 4 times for subsequent use;
(4) experiment establishes 4 process, each process repetition 3 times, and plot area is 6m
2(3m × 2m), each process adopts the group arrangement of random district, and a stress level is established in experiment, and experimental session hillslope processes maintains severe water stress (40% ~ 20% FC), and experimental design is as follows:
1) 1L blank cultures is added
2) add 8g nanometer compost and add 1L blank cultures
3) add 8g nanometer compost and add the complex micro organism fungicide that 1L dilutes 100 times
4) add 8g nanometer compost and add the complex micro organism fungicide that 1L dilutes 4 times
(5) after process, in 4-11 in 2014 month-carry out plant height mensuration, take the mode of random sampling to measure at every turn.
Wherein the concentration of PEG6000 is respectively 5%, 10%, 15%, 20% and 25%.Often increase the concentration of a PEG6000, after thalline increment is stable, will could continues the concentration increasing PEG6000, strengthen step by step and drought-enduring mixing microorganisms flora.Used material: choose full seed, perennial Festuca Arundinacea of uniform size (Festuca arundinacea L.) seed is test materials;
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 impurities removing, cross 2mm sieve, be positioned over ventilation opening place 2-3d, air-dry under natural condition, its physico-chemical property is: pH 7.44, saturation moisture content 0.58mL/g, organic 4.68%, full nitrogen 0.21%, available phosphorus 22.03 mg/kg, full potassium 45.61g/kg.
Claims (5)
1. adopt drought-enduring strengthening active nano garbage compost to improve a method for field sod production drought resistance, it is characterized in that being undertaken by following step
(1) with the soil after sterilizing for matrix carries out lawn plant experiment in cultivation, nanometer compost adopts the mixed mode executed to be manured into soil matrix, and during experiment, substrates quantity is 9400gm
-2, compounded turf matrix thickness is 15 mm, and Festuca Arundinacea grass seeds application rate is 160gm
-2, evenly sow after seed being soaked 24h in matrix top layer, first carry out dark processing, the plant strain growth initial stage after sprouting, every day unifies quantitative water supply, to keep culture medium water regime good, then adds 30-35nm nanometer compost 8g and adds the drought-enduring microbial inoculum of strengthening that 1L dilutes 100 times; Or add 30-35nm nanometer compost 8g and add the drought-enduring microbial inoculum of strengthening that 1L dilutes 4 times; What the drought-enduring microbial inoculum of described strengthening adopted is the method progressively increasing PEG6000 concentration, and the concentration of PEG6000 increases with the gradient of 5%, and ultimate aim is 25%; Described microbial inoculum refers to: bacillus cereus, the preparation of Methionin genus bacillus and rhodotorula glutinis 1:1:1 by volume;
(2), after process, carry out plant height mensuration in the 4-11 month in 2014, take the mode of random sampling to measure at every turn.
2. described in claim 1, adopt drought-enduring strengthening active nano garbage compost to improve the method for field sod production drought resistance, 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) experiment establishes 4 process, each process repetition 3 times, and plot area is 6m
2(3m × 2m), each process adopts the group arrangement of random district, and a stress level is established in experiment, and experimental session hillslope processes maintains severe water stress (40% ~ 20% FC), and experimental design is as follows:
1) 1L blank cultures is added
2) add 8g nanometer compost and add 1L blank cultures
3) add 8g nanometer compost and add the complex micro organism fungicide that 1L dilutes 100 times
4) add 8g nanometer compost and add the complex micro organism fungicide that 1L dilutes 4 times
(5), after process, carry out plant height mensuration in the 4-11 month in 2014, take the mode of random sampling to measure at every turn.
3. described in claim 1, adopt drought-enduring strengthening active nano garbage compost to improve the method for field sod production drought resistance, wherein: the concentration of PEG6000 is respectively 5%, 10%, 15%, 20% and 25%, often increase the concentration of a PEG6000, after thalline increment is stable, the concentration increasing PEG6000 could be continued, strengthen step by step and drought-enduring mixing microorganisms flora.
4. the method adopting drought-enduring strengthening active nano garbage compost to improve field sod production drought resistance described in claim 1 is promoting the application in the green degree anaphase effect of turf.
5. the method adopting drought-enduring strengthening active nano garbage compost to improve field sod production drought resistance described in claim 1 extend the turf green phase, in advance turn green the time, improve application in regrowth rate.
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