WO2017017633A1 - Composition et procédé permettant de favoriser la croissance de plantes herbacées et de favoriser une accumulation de matière organique dans le sol - Google Patents

Composition et procédé permettant de favoriser la croissance de plantes herbacées et de favoriser une accumulation de matière organique dans le sol Download PDF

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Publication number
WO2017017633A1
WO2017017633A1 PCT/IB2016/054515 IB2016054515W WO2017017633A1 WO 2017017633 A1 WO2017017633 A1 WO 2017017633A1 IB 2016054515 W IB2016054515 W IB 2016054515W WO 2017017633 A1 WO2017017633 A1 WO 2017017633A1
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WIPO (PCT)
Prior art keywords
soil
composition
composition according
lactobacillus
microorganism belonging
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PCT/IB2016/054515
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English (en)
Inventor
Giovanni DINELLI
Ilaria MAROTTI
Diana DI GIOIA
Bruno BIAVATI
Francesca GAGGIA
Loredana BAFFONI
Sara BOSI
Mattia ACCORSI
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Alma Mater Studiorum-Universita' Di Bologna
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Application filed by Alma Mater Studiorum-Universita' Di Bologna filed Critical Alma Mater Studiorum-Universita' Di Bologna
Priority to EP16757081.1A priority Critical patent/EP3328203A1/fr
Publication of WO2017017633A1 publication Critical patent/WO2017017633A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom

Definitions

  • the invention relates to a composition and a method for promoting the growth of herbaceous plants, in particular herbaceous plants comprised in turfs for sporting or ornamental use and in agricultural lands, and favouring an accumulation of organic matter in the soil in which the aforesaid herbaceous plants live and grow.
  • Thatch means a layer consisting of roots, leaves and undecomposed vegetable matter, which layer is produced naturally in a lawn that is in good conditions.
  • the thatch plays a key role in the structure of a golf facility's lawn, especially inside the putting green (the grassy area surrounding the holes), since a certain amount of thatch is necessary to reduce the trampling stress and for the impact dynamics of the golf ball.
  • agronomic methods that enable the thatch to be degraded and that are carried out at preset times of the year, like the drilling, the coring, the vertical mowing (vertical cutting of the turf) and the top-dressing (filling of the surface depressions of the turf with soil and sand).
  • the aforesaid treatments aim to remove compacted matter, enable an oxygenation at root level and restore a correct porosity of the soil.
  • a drawback of the known agronomic methods consists of the fact that although the latter are effective they inevitably cause an aesthetic and functional interruption to the golf facility's lawn in which they are carried out.
  • W096/29874 discloses compositions containing the strains YCED 9 and WYE 53 of Streptomyces, which are usable for reducing the susceptibility to fungal infections of plants of agronomic interest.
  • the strains YCED 9 and WYE 53 are able to produce antimicrobial metabolites, including antifungal metabolites, with which it is possible to treat the root apparatuses of the plants or the seeds of the latter.
  • a drawback of the composition disclosed in W096/29874 consists of the fact that the antifungal action is performed by bacterial strains producing antimicrobial metabolites.
  • delivering the aforesaid metabolites to the soil tends to reduce significantly the biodiversity in situ, eliminating microbial species that are potentially useful or at least not harmful to the plants.
  • One object of the invention is to improve known compositions and methods for preventing the thickening of the layer of thatch in the turfs and for containing the fungal infections in the latter.
  • Another object is to make available a composition and a method for promoting the growth of herbaceous plants in the turfs, in particular favouring a beneficial interaction between the endogenous microorganisms of the soil and the root apparatuses of the herbaceous plants, and favouring an accumulation of organic matter in the aforesaid turfs.
  • a further object is to make available a composition and a method for preventing the thickening of the layer of thatch in the turfs, so as to avoid, or at least minimise, the intervention of specialised personnel and the use of specific equipment.
  • Another further object is to make available a composition and a method for containing the fungal infections in the turfs that enable the use of toxic chemical substances and/or of microbial metabolites that are harmful for biodiversity to be avoided.
  • a composition for promoting the growth of herbaceous plants in a soil and favouring an accumulation of organic matter in said soil, as defined in claim 1.
  • a method for promoting the growth of herbaceous plants in a soil and favouring an accumulation of organic matter in said soil, as defined in claim 9.
  • compositions and a method are made available that, as highlighted by the experimental tests conducted (and disclosed in detail below), ensure clear effects on both the plants and on the growth substrate of the latter.
  • composition and the method according to the invention make it possible to obtain:
  • composition and the method according to the invention can be easily and effectively used in the cultivation of both plants of agronomic interest and ornamental plants. In particular, as the intervention of specialised personnel is not necessary, the composition and the method according to the invention can be used in those growth environments that generate significant technical and maintenance problems.
  • compositions and of the method according to the invention can be widely spread in agricultural systems with low input, such as, for example, organic farming, given the organic nature of the composition.
  • the composition according to the invention does not contain xenobiotic substances, which are harmful for the environment and, in particular, for the natural microbiota of the rhizosphere.
  • Figure 1 shows two graphs, illustrating respectively the production of wet leaf biomass and the production of dry leaf biomass in parcels of turf treated with the composition according to the invention and in parcels of turf treated with a known composition;
  • Figure 2 shows two graphs, illustrating respectively the production of wet thatch and roots biomass and the production of dry thatch and roots biomass in the parcels shown in Figure 1 ;
  • Figure 3 is a graph illustrating the tearing strength in the parcels shown in Figure 1 ;
  • Figure 4 is a graph illustrating the organic carbon content in the first 5 cm of soil of the parcels shown in Figure 1 ;
  • Figure 5 shows three graphs, illustrating respectively the nitrogen percentage content, the carbon percentage content and the carbon/nitrogen percentage ratio in the soil of the parcels shown in Figure 1 ;—
  • Figure 6 is a graph illustrating the humidity content in the first 5 cm of soil of the parcels shown in Figure 1 ;
  • Figure 7 is a graph illustrating an evapotfanspiration test conducted on grass sods removed from the parcels shown in Figure 1 ;
  • Figure 8 shows a graph of the RGB colour space and a photograph of a parcel of turf treated with the composition according to the invention, after two months of treatment
  • Figure 9 shows four graphs, illustrating respectively the chlorophyll a content, the chlorophyll b content, the chlorophyll a / chlorophyll b ratio and the total chlorophyll content in leaves of grass (Agrostis stolonifera) removed from the parcels shown in Figure 1 ;
  • Figure 10 shows a series of photographs, summarising the profiles of the soil in the parcels shown in Figure 1 ;
  • Figure 1 1 is a graph illustrating the reduction in the thickness of the layer of thatch in the parcels shown in Figure 1.
  • composition according to the invention comprises the following components:
  • a mycorrhizal fungus namely a fungus the mycelium of which is able to establish a mutual symbiosis with the root of an upper plant (mycorrhizal symbiosis or mycorrhiza);
  • humic acids long chain organic molecules, of colloidal type.
  • the microbial mixture consists of microorganisms belonging to:
  • SB l Candida sp. Forest soil Antagonist-protective action (competitor against pathogenic fungi and bacteria)
  • the action performed by the single strains is defined as "probable” inasmuch as it is not identifiable with certainty.
  • the overall and experimentally verified beneficial effects that are obtainable through the composition according to the invention are therefore attributable to a synergic action.
  • the latter is performed by the various strains of the aforesaid microbial mixture, together with the other components of the composition (mycorrhizal fungus and source of humic acids).
  • the strain LB2 (L. buchneri) was filed on date 13/07/2015 at the MSCL collection ⁇ Microbial Strain Collection of Lithuania) with number P I 482.
  • strain LB6 L. parafarraginis
  • MSCL collection Microbial Strain Collection of Lithuania
  • strain LB 8 (L. diolivorans) was filed on date 13/07/2015 at the MSCL collection (Microbial Strain Collection of Lithuania) with number P I 484.
  • strain LB9 L. plantarum
  • MSCL collection Microbial Strain Collection of Lithuania
  • strain TS l (Stenotrophomonas maltophilia) was filed on date 13/07/2015 at the MSCL collection (Microbial Strain Collection of Lithuania) with number P I 485.
  • strain TS l 2 Bacillus subtilis
  • MSCL collection Microbial Strain Collection of Lithuania
  • strain SB l (Candida sp.) was filed on date 1 3/07/2015 at the MSCL collection (Microbial Strain Collection of Lithuania) with number P I 486.
  • the mycorrhizal fungus of the composition according to the invention is of the arbuscular - vesicular type and belongs to the Glomus intraradices species.
  • This mycorrhizal fungus performs a protective function because it reduces biotic and abiotic stresses to which the herbaceous plants are subjected.
  • the aforesaid mycorrhizal fungus performs an ameliorative function, since, through the arbuscular structures thereof, it increases the capacity of the plants to absorb nutritional elements and the internal translocation of mineral elements.
  • composition according to the invention contains propagules - namely spores - of Glomus intraradices in a concentration of 1 -2 propagules/ml of product.
  • the source of humic acids is the leonardite, a fossil humus.
  • the leonardite is usually used as an organic soil improver and is available commercially in the form of powder or granules.
  • composition according to the invention contains leonardite in a concentration comprised between 1 % in weight and 5% in weight.
  • Example 1 Preparation of a composition according to the invention
  • Each culture medium is prepared and sterilised by using known laboratory methods and apparatuses.
  • Each flask is inoculated with 5% (vol/vol) of a liquid culture of the corresponding strain, grown for a night in MRS broth at a temperature of +37 °C (Lactobacillus buchneri, Lactobacillus parafarraginis, Lactobacillus diolivorans and Lactobacillus plantarum), in BHI broth at a temperature of +30 °C (Stenotrophomonas maltophilia and Bacillus subtilis) or in SDB broth (Candida sp).
  • the (seven) flasks are incubated for 48 h at +37 °C (strains of Lactobacillus buchneri, Lactobacillus parafarraginis, Lactobacillus diolivorans and Lactobacillus plantarum) and at +30 °C for the other microorganisms (strains of Stenotrophomonas maltophilia, Bacillus subtilis, Candida sp.).
  • the mycorrhizal fungus is prepared by starting from a commercially available product, for example Endomic Clay - Altea ® , which is a formulation that is based on spores of Glomus intraradices .
  • the product is in the form of a wettable powder having a spore concentration equal to 10% ( 100 propagules/g).
  • humic acids As a source of humic acids, leonardite in powdered form is used. The latter is dissolved (1 % w/v) in the physiological solution containing the microbial mixture and the spores of mycorrhizal fungus, obtained as disclosed in preceding point 1.2.
  • the microbial mixture, the spores of mycorrhizal fungus and the source of humic acids are mixed reciprocally until a final product is obtained, corresponding to the composition according to the invention.
  • composition according to the invention can be stored for 24 h at +4°C in conditions of semi-darkness, until the moment of use (field application).
  • Example 2 Treatment of parcels of turf with a composition according to the invention
  • the experimental test was conducted on a turf intended for sporting use, namely a professional putting green inside a golf club in the province of Bologna (Emilia-Romagna, Italy).
  • the putting green consisted entirely of the herbaceous vegetable species Agrostis stolonifera and was made according to USGA (United States Golf Association) guidelines of 1994.
  • Each parcel extended over about 2 m .
  • the commercial product which was used as a comparison element in the experimental test, consists of lactic bacteria (1.2 x 10 6 UFC/ml), yeasts (9.5 x 10 4 UFC/ml) and photosynthetic bacteria (1.8 x 10 5 UFC/ml).
  • the treatments were administered to the parcels weekly for a period of two months (from July 10, 2012 to September 1 1 , 2012) and were carried out during the weekly closing of the golf club (Tuesday afternoons), so as to avoid possible dispersion and/or contamination induced by the trampling of the athletes.
  • composition according to the invention was named "DiSTA PLUS" and it is indicated by this name in Figures 1-1 1.
  • the composition according to the invention was sprayed at the leaf level by a pump operating at pressure equal to 3 bar, so as to administer 1 litre of composition to each parcel.
  • the watering system of the putting green was activated for 2 rriin., providing for a supply of water of about 10 mm/m 2 .
  • the composition was washed away from the leaf surface and transferred inside the soil and the thatch.
  • the treated (and untreated) parcels were subjected to weekly analyses (from July 9, 2012 to September 12, 2012) to determine with the maximum possible accuracy the effects of the composition according to the invention on the chemical, physical and biological components of a turf intended for sporting use.
  • the analysis of the leaf biomass was conducted by taking into consideration both wet and dry biomass. Once a week a sod of lawn was taken and leaves were cut therefrom and weighed in laboratory to obtain the weight of the wet biomass. Immediately afterwards, the leaves were placed in a dryer at +50°C for 4 days. When all the humidity contained in the leaf tissues had evaporated, the leaves were weighed again to determine the dry weight thereof, namely the weight of the dry biomass.
  • the soil present was removed by washing, leaving only roots, leaves and the undecomposed vegetable matter that constitutes the thatch.
  • the sods were cleaned inside a sieve having 0.5 mm pores. After determining the wet weight of the vegetable tissues (wet biomass), they were placed in a dryer at 50°C for 4 days. When all the humidity contained in the vegetable tissues had evaporated, they were weighed again in order to determine the dry weight (dry biomass).
  • the tearing strength of the leaves was analysed directly in the field, using a mechanical dynamometer.
  • the end of the dynamometer provided with tweezers, was connected to the base of the culm of an Agrostis stolonifera plant.
  • the resistance provided by the dynamometer pulling in the opposite direction to the soil, caused the plant to tear.
  • Both the commercial product and the composition according to the invention determined an increase in the tearing strength of the leaves of Agrostis stolonifera compared with the control. As shown in Figure 3, the composition according to the invention showed a slightly greater effect than the commercial product.
  • the organic component of the soil was analysed on samples (portions of soil or sods) corresponding to the first 5 cm of soil, starting from the base of the culm of Agrostis stolonifera plants.
  • the samples were first weighed, then dried at 50°C for 4 days, until they dried out completely. The weight at humidity equal to zero was determined.
  • the samples were placed in a muffle at 550 °C for 3 hours, so as to incinerate all the organic component present.
  • the samples, once removed from the muffle were cooled in a container in the absence of humidity and then weighed to determine the weight loss corresponding to the incinerated quantity of organic carbon.
  • composition according to the invention produced a gradual decrease in the organic carbon content in the first 5 cm of soil below the layer of Agrostis stolonifera, whilst the treatment with the commercial product did not show statistically significant differences compared to the control.
  • the difference in values induced by the composition according to the invention is ascribable to the degrading of the thatch produced by the introduced microorganisms, which mineralised and transformed into metabolytes the carbon constituting the cellulose and the lignin of the thatch.
  • both the applied treatments caused an increase in the values of total nitrogen and total carbon inside the first 5 centimetres of soil.
  • the increase of carbon inside the soil was probably caused by the use of products of an organic nature like microorganisms and humic acids, long carbon chain compounds.
  • the humidity of the soil was determined by taking samples (cores) of soil and determining the water content thereof by drying the samples at a temperature of 80 °C, until the stable dry weight was reached.
  • composition according to the invention caused a statistically significant, and equal to 6%, decrease of the humidity in the soil.
  • the analysis of the evapotranspiration was carried out by using sods of soil having 2,5 x 10 cm in area and 8 cm in depth. These sods were taken to laboratory, weighed and then watered until they reached the field capacity (which is understood as the water content in a soil that is associated with a hydraulic potential of -0.33 bar). The sod was almost completely insulated with parafilm, leaving only the upper surface free, so that the water loss was due only to evapotranspiration. The sods were placed in a growth chamber with artificial lighting. The photoperiod and the temperature were adjusted at: 16 hours of light at +28°C; 8 of dark at +20°C. The weight loss of each sod, as caused by the evapotranspiration of water, was measured each day at 17:00 until the soil was completely dried.
  • the sods taken from the parcels treated with the composition according to the invention showed greater ability to evapotranspire the water (contained therein) than the control and the commercial product, losing half the water content in 10 days.
  • the sods treated with the commercial product did not determine a statistically significant difference with respect to the control.
  • the analysis conducted for determining the leaf pigments content inside the vegetable tissues consisted of a quantitative determination of chlorophyll a, chlorophyll b and carotenoids by a UV spectrophotometer.
  • the leaves were sampled from the parcels and immediately frozen in liquid nitrogen to be able to be subsequently analysed in laboratory.
  • the treatment with the composition according to the invention determined a 76% increase in the concentration of chlorophyll a in the first month, decreasing to 48% at the end of the test.
  • a similar trend (Figure 9, top right-hand graph) was also obtained for chlorophyll b, both for the plants treated with the commercial product and for the plants treated with the composition according to the invention.
  • the ratio between the concentration of chlorophyll a and the concentration of chlorophyll b is a typical parameter for describing the physiological state of a plant. Generally this ratio is 3.5-4.5 for plants that have grown in full sunlight and 2.5-3.5 for plants in partial shade (Lichtenthaler, H.K., 1981. Adaptation of leaves and chloroplasts to high quanta fluence rates. Photosynthesis VI. Ed. G. Akoyunoglou, Balaban Internat. Science Service, Philadelphia, pp. 273-287). All the carried out analyses detected a ratio between chlorophyll a and chlorophyll b of about 1.5 -2, which indicates a condition of general turf stress.
  • the composition according to the invention caused a constant reduction in the thickness of the layer of thatch, reducing the latter by more than 26 % compared to the start of the experimental test.
  • the commercial product caused a reduction in the thatch, but of only 6% with compared to the values at the start of the test and anyway without producing a statistically significant effect compared to the control.
  • a further change to the profile of the soil induced by the composition according to the invention consisted of an increase in humus, i.e. the dark brown layer below the thatch and consisting of decomposing organic matter.
  • composition according to the invention produced an increase in the production of leaf biomass. Although it does not comply completely with the required parameters for the putting green of a golf course (in which it is sought to minimise growth to lower the frequency, and thus the costs, of grass cutting), this experimental datum is nevertheless interpretable positively.
  • the growth spurt induced by the composition according to the invention is attributable to a greater availability of nutrients for the plants, which greater availability is obtained owing to the degradation of the thatch. Consequently, a new balance of fertilisations - made less frequent during the year taking into account the recycling of the biomass induced by the composition according to the invention - can reduce the production of leaf biomass and enable the number, and thus the cost, of the maintenance interventions on the turfs to be reduced.
  • the treatment with the composition according to the invention has produced a clearly positive effect on the respiratory capacity of the lawn.
  • This effect is strongly correlated to the reduction of the layer of thatch, which reduction is produced by the metabolic and synergic action of the microorganisms, of the mycorrhizas (formed owing to the presence of the mycorrhizal fungus in the composition according to the invention) and of the humic acids.
  • the reduction of the layer of thatch reduced the insulating effect that the latter causes between soil and surface, facilitating the gaseous exchanges, the infiltration of the water and consequently improving the physiological state of the entire grass surface.

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  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Pest Control & Pesticides (AREA)
  • Biotechnology (AREA)
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  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Soil Conditioners And Soil-Stabilizing Materials (AREA)

Abstract

La présente invention concerne une composition permettant de favoriser la croissance de plantes herbacées dans un sol et favoriser une accumulation de matière organique dans ledit sol, comprenant un mélange de micro-organismes, de spores d'un champignon mycorhizien et d'une source d'acides humiques, ledit mélange de micro-organismes comprenant au moins un micro-organisme appartenant au genre Lactobacillus, au moins un micro-organisme appartenant au genre Bacillus, au moins un micro-organisme appartenant au genre Stenotrophomonas et au moins un micro-organisme appartenant au genre Candida. L'invention concerne également un procédé permettant de favoriser la croissance de plantes herbacées dans un sol et favoriser une accumulation de matière organique dans ledit sol, consistant à administrer à ces plantes et/ou ce sol une composition ; ladite composition comprenant un mélange de micro-organismes, de spores d'un champignon mycorhizien et d'une source d'acides humiques, ledit mélange de micro-organismes comprenant au moins un micro-organisme appartenant au genre Lactobacillus, au moins un micro-organisme appartenant au genre Bacillus, au moins un micro-organisme appartenant au genre Stenotrophomonas et au moins un micro-organisme appartenant au genre Candida.
PCT/IB2016/054515 2015-07-30 2016-07-28 Composition et procédé permettant de favoriser la croissance de plantes herbacées et de favoriser une accumulation de matière organique dans le sol WO2017017633A1 (fr)

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EP16757081.1A EP3328203A1 (fr) 2015-07-30 2016-07-28 Composition et procédé permettant de favoriser la croissance de plantes herbacées et de favoriser une accumulation de matière organique dans le sol

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IT102015000040461 2015-07-30
ITUB2015A002637A ITUB20152637A1 (it) 2015-07-30 2015-07-30 Composizione e metodo per promuovere la crescita di piante erbacee e favorire un accumulo di sostanza organica nel terreno

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111793585A (zh) * 2020-08-12 2020-10-20 杭州新原起生物科技有限公司 布氏乳杆菌及其培养法和在农业种植中的应用
CN111793585B (zh) * 2020-08-12 2023-06-30 杭州新原起生物科技有限公司 布氏乳杆菌及其培养法和在农业种植中的应用

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