FR2880344A1 - Organic or mixed organic-inorganic fertilizer containing stimulatory bacteria, useful particularly for non-leguminous crops, includes bacteria selected from soil for hardiness - Google Patents

Abstract

Preparation of organic and/or organic-inorganic fertilizer (X) containing hardy bacteria (B) that promote growth of plants comprises preparing an inoculum of (B); fermentation and formulation of the inoculum as a solid state product; and combining the resulting powder with the fertilizer. Independent claims are also included for: (1) fertilizer (X) prepared by the new method; (2) any (X) that contains fermented (B), formulated as a solid state material; (3) screening, for hardiness, bacteria (diazotrophes and able to dissolve phosphorus) that promote growth of plants; and (4) determining the level of bactericidal activity of biosolids used to prepare (X). ACTIVITY : Fertilizer. Maize was grown in a podzol (56% sand; 34% silt; 10% clay) in Quebec. When grown with application of a mixed organic-inorganic fertilizer at 3.8, 14 and 0.3 kg/hectare of nitrogen, phosphorous pentoxide and potassium oxide, respectively, the yield was 103% of that for a sample grown without any fertilizers. The same application of nutrients from a fertilizer inoculated with hardy soil bacteria gave a yield of 109%. MECHANISM OF ACTION : None given.

Description

TECHNICAL AREA

  The technical field of the invention is that of the manufacture of bacterial inocula intended for agriculture and, more particularly, the bacterization of crop residues on the ground, residual fertilizing materials and / or organo-mineral fertilizers. The (bio) industrial sectors such as solid and / or liquid fermentation, as well as the formulation and packaging of fertilizers are concerned.

  STATE OF THE ART Hardiness and active persistence of plant growth promoting bacteria (or BFCP), sometimes referred to as edaphic and / or saprophytic competence, or, in English, fitness in soil, and the key factor that can determine the success of an inoculation (bacterization) protocol for soils, crop residues on the soil, roots, seeds and, in the context of the present invention, organic and organo-mineral fertilizers (EO / EOM). However, this hardiness is a poorly understood phenomenon given the complexity of the biological processes that underlie it. Its direct and positive demonstration using biomolecular techniques, which often presuppose gambling and the expression of relatively simple mechanisms, does not seem feasible. However, the agronomic and bioindustrial importance of the BFCP remains, and the usefulness of techniques that can promote its expression and / or selection (obtaining and retention) is undeniable. It is therefore desirable to make available a technique making it possible to retain the most hardy BFCP strains in mother cultures of telluric bacteria (CMBT) and therefore, in principle, the ones best suited to life in arable soils.

  In addition, the present inventor has developed an additional agronomic and bio-industrial valorization of these BFCP biomasses particularly well adapted to life in soils, as well as a simple analytical method capable of revealing the dominance and the survival of this type. of bacteria despite the bactericidal environment proximal to EOM enriched with mineral fertilizing salts.

  The present invention makes it possible, inter alia, to manufacture and agronomically use EOMI bacteria (I for inoculants).

  Fertilization of crops with nitrogenous, phosphatic and potassium fertilizers of chemical syntheses, or NPK (Nitrate Phosphate Potassium), is an important advance in agriculture but is today associated with many agro-environmental concerns. that EO / EOM avoid, and more particularly, the EOMI (ie EOM + inocula BFCP) bactericized as proposed here. First, the marginal agronomic effectiveness of NPK decreases as the yield increases attributable to them increase. This causes clogging of NPK soils, thus increasing the risk that these highly soluble minerals will be transported to the groundwater thus contributing massively to diffuse pollution of agricultural origin. Secondly, this high solubility of NPK fertilizers in itself poses a certain risk of agri-environmental pollution. These characteristics of NPK fertilization of arable crops contribute in part to increasing the risk of pollution of soils, but especially of water, by nitrates and phosphates.

  In order to reduce the agro-environmental impacts of nitrogen and phosphate fertilizers, there is in principle a range of intervention measures. Without listing them all, it is generally better to regulate the use of NPK fertilizers, particularly by promoting their uptake by crops by modifying their in situ chemistry and / or by better assessing the current needs of the crop in ways that to avoid overdoses.

  In this sense, bacteriization of EO / EOM is desirable and agronomically useful. Indeed, it makes it possible, on the one hand, to confer a triple organic, mineral and bacterial action on starter fertilizers, and, on the other hand, facilitates the conditioning and the transfer towards the targets of the BFCP inocula, in particular with regard to non-leguminosea crops, abandoned by the bio-industrial sector of microbial inocula for agriculture.

  This microbiological evaluation of the EOMs therefore makes it possible to significantly reduce the amount of phosphorus introduced at the start of the crop. Indeed, EOMs that are bactericidal or not, are necessarily less concentrated in P than conventional starter fertilizers; in equivalent hectare doses, the EOMs therefore reduce the intake of P at startup and thus allow the user to delay the supply of organic fertilizer supplements, biosolids or even slurry during the season, while respecting, and this is essential here, the NPK mass balances that are dictated by possible agri-environmental fertilization plans (PAEF), and / or any other agri-environmental regulation. At equal efficiency, the EOM bacteria according to the present invention can significantly reduce the quantities of phosphate inputs required to start crops, thus allowing an environmentally friendly recovery by avoiding other organic materials, manure, manure, biosolids, Fertilizing residual materials (FRM), etc., whose disposal and management are often problematic.

  So called biosolids is any organic matter, which looks like black earth, resulting from the treatment of wastewater. Unlike sewage sludge, biosolids have been treated to reduce or eliminate pathogenic organisms they may contain. Biosolids are rich in nutrients needed for crops, and they are often used as fertilizer for farmland. We will then talk about spreading biosolids.

  Biofertilization using EO / EOM and inocula BFCP the state of the art According to Parent (2003), EOMs are not just a mechanical mixture of granules of organic and inorganic matter. Mineral and organic materials must be premixed and granulated together to increase nutrient content and granule density (according to Parent et al., 2000, for example). The minimum required content is 7.5% carbon in the composition of the granule; those which do not contain exogenous mineral substances are therefore called organic fertilizers (EO) granules. In principle, and according to Parent (2003), the interaction between the organic ligands (organic matter) of EO / EOM and the fixation sites normally occupied by orthophosphate (phosphate) in the soil makes it possible to improve elementary efficiency. For the most part, these binding sites are iron and aluminum hydroxides as well as calcium. By occupying these binding sites, these ligands indirectly increase the availability and therefore the assimilation of orthophosphates by the plant.

  The biofertilization of Leguminosea cultures by inoculation of BFCP is already well known and has been the subject of a major bio-industrial development (Bashan 1998). This is far from the case for non-Leguminosea crops. The agronomic efficacy of BFCPs for non-leguminous crops is generally severely limited by the amount of root exudates produced during the season (Lynch and Whipps 1990, Alden et al., 2001). In the presence of large quantities of crop residues, this limitation can be circumvented insofar as the BFCPs are applied directly by spraying on crop residues, which are thus valued as carbon and energy substrates (Claude and Fillion 2004, Claude 2001, Claude 2003 and FR 2833016). In the absence of such amounts of crop residues, the problem remains, and the agronomic utility of BFCPs for non-Leguminosea crops limited.

  Traditionally, and in the absence of abundant residues of cellulosic culture at the seedlings, the bacterization-BFCP of large non-leguminous cultures is done by seed lamination with an inoculum designed for this purpose (Bashan 1998). Such bacterization of seeds can be problematic given the bacteriocidal nature of certain chemical compounds used for the phytosanitary treatment of seeds. More generally, the bacteria used for the bacterization of non-Leguminosea seeds lack edaphic competence and are relatively poorly adapted to life in soils (Van Veen et al 1997).

  It is therefore useful today to carry out this biofertilization by bacterization by alternative routes, in particular, via the bacterization in the fall of crop residue on the soil, if there is (Claude and Fillion 2004; Claude 2001; Claude 1997). In the absence of such residues, the present invention also allows the bacterization of EO / EOM using bacterial inocula advantageously produced, formulated and packaged in the solid state, incorporated into the EO / EOM either by way of laminating, either by integrating these inocula directly to the premix during their manufacturing processes.

  Starter fertilizers Localized fertilization consists in applying a part of the fertilizer close to the seed at the time of sowing, or at the beginning of the crop. This practice is advantageous in soils that are poor in assimilable phosphate and / or highly acidic, where phosphorus is not very mobile in the soil and easily immobilized by iron and aluminum compounds (CPVQ 1988). These fertilizers, or so-called starter fertilizers, are generally of two types (Craaq 2003): based on mono- or di-ammonium phosphate (MAP and DAP, respec- tively), or based on urea (Alkanani and MacKenzie 1996 ).

  In the first case, the aim is to reduce the fertilizer / soil contact so as to reduce the degree of physicochemical fixation on the oxy-hydroxide sites of iron and aluminum, and in the second case the degree of volatilization of urea in the form of ammonia. In general, acid-base reactions and osmolysis phenomena in the vicinity of granules containing N and P minerals may interfere with the viability of reintroduced BFCPs in their proximities. Given the high organic matter content of EOMs, a carbon content detrimental to their weight efficiency compared to NPK fertilizers, the immediate reactivity of the granules is reduced and the mortality of BFCP possibly reintroduced in their vicinity is increased.

  In Quebec and Canada, the market for phosphate starter fertilizers initially seems to be the most promising for EOMI (EOMI). From 10 to 15 million units (i.e. kg P205 / ha) are sold in Quebec and over 20 million in Ontario, or about one quarter of the phosphate fertilizer units normally consumed on the market; the Canadian market as a whole is in the order of $ 125 million CDN. The French market amounts to over 150 million units. The replacement value based on the 1g price of P205 is approximately $ 1.00 CDN. For the most part, the lack of a weight efficiency comparable to that of NPK starter fertilizers, the use of EO / EOM remains marginal today. It is precisely this weight efficiency that improves the present invention.

  Biofertilization using EO / EOM bacteria with BFCP It would be advantageous to combine the application of P starter fertilizer and inocula BFCP. In fact, the accumulation of phosphorus in reaction with calcium, iron and / or aluminum in the soil is a marked phenomenon in many agricultural soils (Beauchemin and Simard 1999, Zhang et al 1995, Giroux et al. Pellerin 2003). This accumulation leads to an increase in the pollutant load of these soils, and therefore represents a serious agro-environmental problem in several cases (Khiari et al., 2000). In addition, soils rich in phosphorus with little or no solubility, both in France and in Quebec, respond poorly to fertilization - P (eg Chabot et al 1998, Craaq 2003). Thus, on soils with M3 indices (P / AI) above certain pre-established critical values for this type of soil, the probability of response to phosphate fertilizers is generally low (for example, less than 20% according to Craaq 2003). . The invention makes it possible to improve this response.

  In so-called P-rich soils, the efficiency of P fertilizers is therefore limited although there is no indication that the crop's full yield potential has been reached. The valuation of P and more particularly of P fertilizers on these soils is therefore incomplete. In general, the agronomic efficiency of P fertilization on these soils, which are widespread in many temperate agricultural regions in both Quebec and France (Beauchemin and Simard 1999), can be partially improved using two techniques: - the use of EOM (Khiari and Parent 2003, Parent 2003); the bacterization of rhizospheres using BFCP (Antoun et al., 1998, Chabot et al., 1996).

  These two techniques, on the other hand, currently suffer from several technical problems which limit their efficiency and therefore their deployment and use by the farmer. The agronomic efficacy of BFCP inocula is undermined by: insufficient carbon near the roots (Lynch and Whipps 1990, Alden et al., 2001), some bacteriocidal properties of certain plant protection products and insufficiently sought edaphic competence (Van Veen et al. . 1997). The agronomic efficiency of EOMs, and especially EOs, is limited by poor weight efficiency given their organic matter content, which is about five to six times their P2O5 content (Khiari and Parent 2003). The invention proposes to combine these two strategies. It is also possible to overcome these limitations, especially when used as a starter fertilizer.

  o However, the valorization of EO / EOM by bacterization-BFCP has so far been limited because of certain technological obstacles, including the incompetence of the BFCP strains used in soil life, especially in the presence of strong osmolarities in the vicinity of the soil. starter fertilizer, the difficulty of obtaining low solid formulations thereof, and finally the relatively high costs associated with filming ultimate MRF.

  The present invention therefore proposes to solve these problems, while allowing the manufacture and economical use of EOMI (i.e. EOM + inocula BFCP). This was possible thanks to a double contact between the BFCP cells. Indeed, the first contact makes it possible to boost the growth kinetics of the selected BFCP cells as constituents of the eventual solid inoculum, and the second makes it possible to ensure the packaging and shipping of these cells to the targets in situ. including organo-mineral fertilizers (EOM) for crop start-ups (Figure 1). The invention thus makes it possible to ensure the production of an EOMI (i.e. EOM + inocula BFCP) with a triple action, organic, mineral and bacterial.

  Many agricultural soils, especially during spring planting, have little or no crop residues on their surface. Soils low in crop residues at planting time are also not conducive to biofertilization of non-Leguminosea crops using BFCP inocula. Indeed, given the lack of effective symbiosis between the BFCP and the non-Leguminosea culture, said BFCP cells would benefit from using the residues as carbon substrates and energy, which in this case is impossible. Although in such situations, the contribution of exogenous crop residues by the farmer is unthinkable agronomically speaking, the contribution of soil amendments is, on the other hand, advisable. However, the required hectare doses are generally of the order of 10 Mg per hectare or more (N'Dayegamiye 2000), and the bacterization BFCP of such quantities of material is not economical and inefficient because of the very large amount of associated bacterial cells that are able to mask the in situ action of reintroduced BFCPs. The bacterization of a smaller quantity of amendments in the form of biosolids, biomasses, fertilizer residual materials, and more particularly of EO / EOM, must therefore be considered. It is necessary to place these materials and BFCPs to reintroduce near seeds using a combined seeder, as for starter fertilizers. The use of EO / EOM for the start of non-Leguminosea agronomic crops as support and shipping for the said BFCP is therefore particularly advantageous.

  The present invention therefore particularly relates to in situ bacterization of non-leguminosea crops when there is little or no crop residues on the soil at the time of planting.

  PRESENTATION OF THE INVENTION Technical problem Although inocula BFCP and EO / EOM exist separately, it would be advantageous to combine them in a single product to facilitate agronomic use. However, and to date, this combination has been impossible, on the one hand, given the absence of rustic and truly adapted BFCP cells to soil life and able to withstand some osmotic stress, and, on the other hand, because of the unavailability of hard-shell solid inocula BFCP inocula formulated economically by fermentation in the solid state. Really telluric BFCP cells and therefore sufficiently hardy to survive the physicochemical stress of such an addition are therefore indispensable here. However, the implementation of FR 2 833 016 (Claude 2001) makes it possible to obtain this type of cells, which are not only rustic and well adapted to life in the soil, but also able to better withstand physico-chemical stress. in the proximal presence of the fertilizer granules, even of EO / EOM (although in this case the stress is less). In addition, solid formulated and economically produced inocula are needed to make the bacterization of EO / EOM economically viable. However, in the context of the present invention, it has been developed a fermentation method and solid state formulation, or FFES, enhancing the edaphic competence of bacterial cells obtained according to FR 2 833 016. To do this, fine soil particles are introduced in addition to the cellulosic substrate normally present. Indeed, it is known that it is especially the fine particles, advantageously less than 200 micrometers, which are most associated with soil bacterial biomasses (Mills 2003, Jocteur-Monrozier et al 1991, Chenu 1993). In addition, the presence of these fine soil particles during the FFES can not only enhance the edaphic competence of the BFCPs isolated according to FR 2 833 016, but also contribute to it. Indeed, according to Van Dyke and Prosser (1998 and 2000) it is possible to induce the hardiness of already isolated telluric bacterial cells by putting them in contact with inert, organic and inorganic materials, essentially lacking in nutrients. Other technical problems also prevented this combination of EO / EOM and inocula BFCP.

  Firstly, the bacterization of NPK fertilizers could not be achieved because the chemical reactions near the granules interfere with the viability of the proximal BFCPs. Indeed, osmolarity (i.e. the sum of osmotic pressures attributable to all chemical species in solution) near the granules is a determining stress that must face the BFCP cells incorporated in the possible IMO. Studies show the predominant role of the exopolysaccharide (EPS) envelope or glycocalyx, soil bacteria in controlling the diffusion of ions, water, carbon substrates, etc., and therefore likely (well). that not unequivocally demonstrated) in the resistance to a possible osmotic shock caused by a too high immediate concentration of fertilizing mineral salts. That said, there are nevertheless some publications on the osmo-regulating effect of alginates (Alg) produced by certain BFCPs of the genus Pseudomonas, without the saline concentrations being as high as they are in the vicinity of said granules (Hart et al 2001, Chenu and Roberson 1996, Schnider-Keel et al 2001, Miller and Wood 1996, Hartmann et al 1991). For example, according to Schnider-Keel et al. (2001), a maximum salt content (NaCl) of 0.6 M is sufficient to severely attenuate the growth of Pseudomonas; in the case of an EOM enriched with 25% of MAP salts, the molarity of the MAP is about 3.00, or five times the molarity of the NaCl used by Schnider-Keel et al. (2001). In fact, this molarity of 0.6 M NaCl is probably much higher than that in the soil solution. It remains, therefore, that the osmotic stress in the vicinity of enriched EOM granules is considerably stronger than it is in the soil solution. The inventors estimate that the molarity of the consequent salts (ie ammonium, phosphate, is sodium, chlorine, etc.) in the vicinity of an EOM granule enriched with 25% MAP is approximately 3, ie 30 to 60 times that of it is in the soil solution (see Miller and Wood 1996). It is therefore questionable whether the BFCP cells incorporated in the IMOs can survive and remain active and effective despite this stress, at least the time needed for it to dissipate, ie approximately a period of 6 to 10 days.

  Second, adding BFCPs directly to EO / EOM during certain types of manufacturing processes (Parent et al 2000, Marcoux et al., 1992) is impractical, given the nature of these processes that typically involve high temperatures and high temperatures. significant hydrostatic pressures necessary for the cooking and consolidation of extruded granules, temperatures and / or pressures in principle contrary to any bacterization. It should be noted here that the resistance of bacteria to certain thermal shocks is related to their resistance to hydrostatic pressures (Aertsen et al., 2004); especially for the aforesaid manufacturing processes.

  Third, this type of bacterization is also limited by the use of BFCP cells poorly adapted to life in soils (Van Veen et al., 1997) and therefore unable to persist over time and to use the carbon source that represents the organic fraction of EOM.

  Finally, fourthly, it is desirable to economically produce these BFCP, and preferably by solid state fermentation so as to obtain a solid formulation, thereby reducing the shock of bacteria at the time of their combination after contact with the EO / EOM.

  In this sense, a previous patent application (WO 02/057862 A2) requires, for example, for the incorporation of microorganisms into biosolids granules, that these be incorporated sequentially into one of the multiple layers encapsulating said granules. This approach can be limiting and cause problems for non-spherical granules or for granules having multiple layers encapsulating a certain nucleus. There is also a French technology (FR 2 615 203) for the manufacture of an inoculum in the form of granules comprising a core consisting of a feeder matrix coating the microorganisms to be inoculated. However, this technology is not transferable to the manufacture of EO / EOM and / or their precursors biosolids; it does not therefore directly concern the present invention.

  However, the bactericide of these granules, even if they are EOM type, is little or not documented in the scientific literature. Neilsen et al. (1994) speak of the utility of bringing bacteria and MAP together with an organic amendment for the establishment of horticultural crops, without however raising the question of bactericidal MAP at this stage. Gaind and Gaur (2002) note that the contribution of Pseudomonas striata to ash of mining origin, up to 40 tonnes per hectare, is not bactericidal, despite the presence of phosphate fertilizers in the mixture. The hectare doses of ash effectively make it possible to dilute the fertilizer P, avoiding it to interact too directly with the BFCP. Finally, Lima and L. (1996) report the effect of phosphate fertilization on bacterial populations, but for the whole soil, without being particularly concerned about the environment in the vicinity of the fertilizer granules. The bactericidal nature of the proximal environment of EO / EOM granules, or even MAP / DAP granules, has not been described.

  Technical solution: Double contact biosolid-BFCP The present invention advantageously combines inocula BFCP and EO / EOM, creating a new fertilizer material with triple action, mineral, organic and bacteriological, very advantageous for the start of crops not -Leguminosea, especially on soils with a medium degree of phosphorus saturation and / or with little or no crop residues on the soil at planting. The invention makes it possible to better valorize BFCPs that are particularly well adapted to life in the soil and to make them resistant to high osmotic pressures, as well as to the organic carbon of EO / EOM. The invention also makes it possible to solve certain bio-industrial problems which hinder the full development of biofertilization.

  The invention relates to a bioprocess incorporating EO / EOM granules and bacterial biomasses of telluric origin which are particularly well adapted to life in soils. The resulting biofertilizer can be used as a substitute for chemical fertilizers normally used in agronomic crops as a starter fertilizer.

  The method according to the invention comprises the co-formulation of EO / EOM granules and / or their organic precursors with BFCP bacteria capable of solubilizing soil phosphorus (Chabot et al, 1993, Rodriguez and Fraga 1999). This increases the production of the fertilized crops and adds value to the EO / EOM, making them more competitive with NPK fertilizers. This promotes the flow of NPK stocks and FRs.

  The combinations of EO / EOM and BFCP show synergistic activity, allowing to biofertilize non-Leguminosea crops in the absence of soil culture residues that could act as carbon substrates, to make the soils normally perceived as saturated with phosphorus. more conducive to phosphate fertilization and increase their yield.

  Hardiness and edaphic competence (i.e. adaptation to life in soils) of telluric bacteria of the BFCP type and isolated from arable soils according to FR 2 833 016 (Claude 2001) can now be doubly valued. A first time during said fermentation and solid state formulation, or FFES, an integral part of the present invention, and this through a first contact of these bacteria and fine solid particles of soil capable of boosting production bio-industrial of these microorganisms.

  A second valorization of this hardiness and this adaptation to the life in the grounds is possible by a second putting in contact of the thus produced cells FFES and EO / EOM.

  It should be noted that these contacts are either ineffective or harmful to bacteria isolated from arable soils conventionally. Double contacting is therefore imperative with competent telluric bacteria isolated according to FR 2 833 016. In fact, without the contact between the fine soil particles and these bacteria, the cellular title of the inoculum reached by fermentation at solid state would not be high enough to make economically viable the bacterization of fertilizing waste materials such as EO / EOM. In addition, without this first contact, the 10 cells / g should be produced via a more conventional liquid-state fermentation; the freeze-drying and / or drying of the bacterial biomasses thus produced would, however, result in such a mortality that it would be necessary initially to produce not more than 10 but more than 1 viable cells per gram, hence an appreciable extra cost. Finally, the application of bacteria formulated in the liquid state causes a chemical reaction of the EO / EOM at the expense of the survival of telluric bacteria thus placed on their surface.

  In addition, the need to enrich EOMs with a certain amount of mineral salts, especially MAP, would in itself cause the death, by osmolysis, of BFCP cells that are not sufficiently hardy and adapted to life in soils. However, the BFCP cells isolated according to the method described in FR 2 833 016 are particularly hardy and resistant to this type of osmotic stress, which allows them to be advantageously combined with relatively mineral-rich EOMs. In the opposite case, and as soon as banded EOMs with conventional BFCPs are banded, the chemical reactions in the vicinity of said granules will be too bactericidal to allow survival and in situ activity of these organisms.

  Benefits EO / EOM Synergy: The carbon requirements of the BFCPs near the roots are only partially satisfied by the exudates io from them. The abundance of carbon contained in EO / EOM helps to further meet the nutritional needs of these microorganisms. Carbon and organic matter brought near the roots by the EO / EOM as a starting fertilizer are therefore doubly valued; they allow a greater availability of phosphates by their fixation on the fixation sites otherwise occupied by phosphates (Parent 2003), and they act as nutritive substrates for the BFCPs thus reintroduced on the ground near the roots.

  Triple (Bio) Fertilizer Action: The mineral action results from the supply of nutrients incorporated in the EOMs according to, for example, a method described in US Patent 6,056,801. Various EOM (Labrie 2004, Dupuis 2004) and some organic fertilizers (EO) (Provencher 2003) have already been the subject of some development. The organic action results from the interaction between the organic ligands (organic matter) of EO / EOM and the attachment sites normally occupied by orthophosphate (phosphate) in the soil (Parent 2003). For the most part, these binding sites are iron and aluminum hydroxides as well as calcium. By occupying these binding sites, these ligands indirectly increase the availability and therefore the assimilation of orthophosphates by the plant. Finally, the bacteriological action is the result of the interaction between the BFCP near the roots. This interaction can be multiform and involve a number of mechanisms, including diazotrophy, production of growth factors, the dissolution of certain organic and inorganic compounds so as to make them more assimilable by the plant, etc. (see Arshad and Frankenberger, Jr. 1998, Antoun et al, 1998, Chabot et al, 1993, Rodriguez and Fraga 1999).

  Triple-action fertilizers - inorganic, organic and bacterial - are particularly useful because, on the one hand, the synergistic action between the organic component of EO / EOM and BFCP, and, on the other hand, the possibility of biofertilize non-leguminous cultures even in the absence of crop residues that may otherwise have served as carbon substrates. In addition, this triple action makes the soils normally perceived as o saturated with phosphorus more conducive to a certain phosphate fertilization, and therefore to an increase in their yield potentials.

  From an agronomic point of view, the advantages of the invention are the following: reduction of the risks of agro-environmental pollution and improvement of the mass balance sheets of organo-mineral fertilization; simultaneous introduction of inocula BFCP and starter fertilizer, and therefore greater ease in the use of inocula BFCP for non-leguminosea cultures; partial flow of fertilizer residuals (FRs) and biosolids during the production of these value-added inputs for field crop production; - alternative to BFCP bacterization of seeds for non-leguminous cultures sown in the absence of crop residues on the soil; flexibility of use on the farm, and / or incorporation into EOMs of BFCPs at the plant.

  From a bio-industrial point of view, the FFES, an integral part of the present invention makes it possible to reach cell densities of the order of 10 cells per gram of organomineral mixture, advantageously mixed cellulosic culture residues. to fine particles of arable soils, within three to four weeks after the start of incubation, and that from an initial cell density of about 1e7, or sometimes as low as 1e6, or even 1e5 per gram of mixing, under non-sterile conditions and at room temperature if necessary. The bio-industrial advantages of this process, especially for the production of diazotrophic bacteria, are: - A strong reduction of the fermentation capacity requirements, since most of the increase in the cell density of the inoculum is in the medium solid; Suitability of the carrier, mixture of crop residues and soil particles, for telluric bacteria to be reintroduced into the soil. The possibility of using a solid formulation for spraying purposes, which has traditionally been is lacking in bacterial formulations based on peat, or advantageously for EO / EOM film coating; Given the hardiness of candidate BFCPs, their use as a bacterial additive for EO / EOM enriched with mineral nutrients ensures some numerical dominance at the expense of contaminating and less hardy bacterial strains. The degree of microbiological purity of the EOMI thus produced is thus de facto ensured and all the more compatible with the current ecotoxicological regulations.

  Finally, it is important to note that the invention allows to boost, in the context of a certain fermentation in the solid state, the growth kinetics of truly telluric bacterial cells at the expense of those which are not. The invention is therefore perfectly compatible with the state of the art in the field of solid state fermentations, in particular and by way of non-limiting example, according to the technology described in patent EP 0 406 103 A1. In such situations, the production of truly telluric BFCPs isolated according to FR 2 833 016 is boosted within the production times normally required by said solid state fermentation technologies according to the state of the art.

  The invention thus makes it possible to produce, from a choke easily produced by conventional fermentation and an inexpensive organo-mineral mixture based on crop residues and soil particles, all containing only 1 6 cells per gram. a solid inocula rich at 5e9 cells per gram, or even up to 1 e10, so able to bring more than 1 e13 cells per hectare for a hectare dose of about 1 kg.

  More specifically, the present invention thus relates, according to a first object, to a process for the manufacture of organic fertilizer and / or organomineral fertilizer (EO / EOM) which is bacteriostated by rustic plant growth promoting bacteria (BFCP) comprising the following steps: preparation of a BFCP inocula; ii. fermentation and formulation of said inocula in the solid state (FFES); iii. contacting the powder obtained in ii with the EO / EOM.

  According to the present invention, rustic or hardiness of bacterial strains is understood to be their edaphic competence or fitness in soil in English, that is to say their ability to adapt to life in arable soils. This property thus confers said bacteria to survive actively and not in dormancy, and this despite the presence of various stress, such as osmotic stress, water, heat, biological (predation), etc.. Preferably: in step i. BFCPs are obtained from bacterial biomasses of telluric origin; the inocula of BFCP is obtained by a process according to FR 2833016 incorporated herein by reference; more specifically, this method is characterized in that it comprises the steps of: - bringing an aqueous suspension of soil into contact with a matrix substrate, so as to form a hydrated zone, or biofilm, on the surface of the substrate, analogous to that associated with the soil CAH, essentially sheltering the autochthonous bacterial component of the soil, in order to retain at the surface of the matrix substrate, most of the endogenous bacterial microflora in the original edaphic media; maturation of the biofilm with establishment of oligotrophic conditions allowing the most constitutive cells of the edaphic biofilm which have colonized the matrix substrate, to migrate to the supernatant liquid phase the matrix substrate, and gradually to dominate it; recovery and cultivation of the most prolific bacterial strains in rich liquid cultures, native, rustic and persistent in situ, cultivable in liquid media, and therefore able to be produced industrially.

  the BFCPs are chosen from the families Azotobacteracea and Pseudonomacea, in particular the bacteria are diazotrophic and capable of dissolving phosphorus (MPS).

  the bacteria express the AZM gene.

  - the FFES of step ii. comprises contacting the BFCP with a substrate / carbon support and soil particles.

  the substrate / carbon support comprises residues of cereal crops such that the residues of cereal crops consist of particles of 1 to 4 mm, in particular from arable soils and more particularly from analogs of the functional fraction of the clay-humic complex of the ground.

  the soil particles have a diameter of between 50 and 100 micrometers.

  - The moisture content of the powder obtained after ii is between 10 and 25%, more particularly between 12 and 15%.

  the powder obtained at the end of stage ii has a titre greater than 5 × 10 9 / g.

  the dose hectare is between 1 and 5 kg of the powder obtained in ii.

  - bringing into contact iii. is carried out by filming EO / EOM with the powder obtained in ii or by incorporating the powder obtained in ii into EO / EOM granules.

  the composition obtained at the end of step iii has a cellular titre greater than 5 × 10 7 cells / g.

  the powder obtained in II and the EOM are mixed in a ratio of between 1 and 10% by weight, according to the organic matter content of the EOMs. Step iii is carried out without the addition of water.

  the process does not comprise a bactericidal step.

  - EO / EOM comprise at least about 20% carbon.

  EO / EOM comprises 25 to 30% of mineral fertilizing salts, such as N, P, K, ammonium phosphate (MAP).

  the EO / EOM are chosen from organic and organo-mineral fertilizers comprising biosolids, biomasses or other residual fertilizing materials.

  According to another subject, the present invention also relates to an organic or organo-mineral fertilizer (EO / EOM) bacterized comprising plant growth-promoting bacteria (BFCP) fermented and formulated in the solid state that can be obtained by the process of the invention defined above.

  According to another subject, the present invention relates to an organic fertilizer or organo-mineral (EO / EOM) bacterized comprising plant growth-promoting bacteria (BFCP) fermented and formulated in the solid state.

  Preferably, the EO / EOM is as defined above and / or the BFCP is as defined above.

  According to another object, the present invention also relates to a method for the fertilization of cultures comprising the application of EO / EOM bacterized according to the invention.

  Preferably: the cultures are non-leguminosea.

  - the application of the bactericidal fertilizer is carried out as a starter fertilizer.

  - the application of the bactericidal fertilizer is carried out in strips near the seeds.

  the soils are devoid of large amounts of crop residues at the time of sowing.

  According to another subject, the present invention also relates to a method for screening plant growth-promoting bacteria (BFCP) according to their hardiness, said BFCP being diazotrophic and MPS, comprising: - culturing precultures of BFCP at room temperature in a liquid medium devoid of nitrogen and phosphorus with the exception of hydroxypathy, - the application of osmotic pressure, - the incubation of cultures for a sufficient period of time, - the measurement of the optical density (OD) middle at two instants (DO1 and o DO2); - the hardiness of the bacteria depending on the difference (D02-D01). Preferably: - said increased hardiness reflects the expression of the AZM gene in said BFCP.

  - the osmotic pressure is such that it causes the death of the less rustic bacteria but not that of the most rustic bacteria.

  the osmotic pressure depends on the monoammoniacal salt concentration of phosphorus.

  the osmolarity of the mono-amoniacal salt of phosphorus is between 0.26 and 2.8, more preferably between 0.58 and 2.8, and particularly 1.26.

  - said culture time is 28 days.

  - DO1 and D02 are measured respectively at the 3rd and 28th days of culture.

  the ambient temperature is a temperature of between 20 and 22 C.

  - said precultures are diluted to 1 / 5th; 1 / 7th or 1 / 9th.

  Finally, according to another subject, the present invention relates to a method for determining the degree of bactericidal biosolids useful for the manufacture of EO / EOMI bacteriocussed by plant growth promoting bacteria (BFCP) comprising: - culturing of precultures of BFCP in a liquid medium in the presence of said biosolids, - incubation at room temperature, - the measurement of the optical density (OD) of the medium at two instants (OD1 and DO2), - the bactericide of said biosolids being high when the difference (DO2-DO1) is small or vice versa.

  Preferably, said biosolids are selected from envirograin, lisox, lisox C, crumb, GSI, Shoc.

  The invention thus relates to compositions for the bacterization of organic fertilizers (EO) and of fertilizer-organo-minerals (EOM) granules for non-Leguminosea agronomic crops, as well as their manufacturing process. Their manufacture consists of a double contact between: (1) fine particles of soil and plant growth promoting bacteria (BFCP), bacteria isolated because of their edaphic competence, in the context of a fermentation and formulation to the solid state (FFES), and (2) the inoculum BFCP thus produced and EO / EOM, and / or their precursors during the manufacture of said EO / EOM, which can be used as expedients and carbon substrates .

  The invention ensures that these contacts are economical and weakly bacteriocidal despite a content of mineral fertilizing salts of EOM thus formed up to 25 to 30%. The invention makes it possible to confer on these new fertilizing materials a triple mineral, organic and bacteriological action. This triple action allows it to surpass in terms of agronomic efficiency the current EO / EOM when used as a starter fertilizer for non-Leguminosea agronomic crops. This invention is applicable to various farming methods, including the application in band of starter fertilizer. The invention is advantageously implemented in agronomic situations where there is absence of crop residues on the ground at the time of sowing and / or on soils moderately saturated with phosphorus and thus little or not conducive to conventional phosphate fertilization.

BRIEF DESCRIPTION OF THE DRAWINGS

  Figure 1: Schematic representation of the process according to the invention comprising a first and a second contacting of bacteria (BFCP) with (bio) solids to ensure full appreciation of the edaphic competence and the hardiness of bacterial strains particularly well adapted to life in arable soils lo advantageously isolated according to the protocol FR 2 833 016 (Claude 2001).

  Figure 2: Azotobacterial cellular titers of microcosms containing only residues of bacterialized cultures and residues of bacterialized cultures coated with fine soil particles; note the increase in cellular titers attributable to the induction of telluric cells (AZM) by fine soil particles (SPF).

  FIG. 3: Evolution of Azotobacterial Cell Titres in Microcosms Containing Bactericidal Culture Residues Coated with Fine Particles of Soils According to the Invention.

  Figure 4: Dominance of the AZMs according to the iMPS index and as a function of the concentration of nitrogen (a) and phosphorus (b) directly from the MAP in relation to the total N and P content (total) of the granule.

  BEST MODE FOR CARRYING OUT THE INVENTION The present invention comprises: a method of manufacturing mineral fertilizers; - a fermentation process and solid state formulation (FFES) innovative BFCP and isolated from these CMBT, advantageously according to Claude (2001); a method of biofertilizing non-leguminosea cultures by banding in the vicinity of the EO / EOM seeds thus bacteriocussed, advantageously on soils with little or no crop residues on the soil at the time of sowing and moderately saturated with phosphorus according to recognized indices.

  The invention also uses an innovative analytical method to confirm the expression of the rustic phenotypes on the part of the candidate BFCPs concerned. This innovative method presented for the first time in the context of the present invention does not require the intervention of biomolecular and / or bioinformatic techniques, techniques essentially unsuitable for the detection of phenotypes whose genomic and metabolic bases are presumably very complex (see Rüberg et al., 2003), and therefore difficult to reduce and explain via simple adnique transcription mechanisms in proteomes.

  The invention is realized in two stages. First, biosolids / BFCP bacteria must be contacted for the first time in order to manufacture the BFCP inocula to be used for the bacterization of the EO / EOM, preform post-constitution of these. In a second step, a second contact between biosolids and BFCP bacteria now formulated in the solid state must be carried out in order to definitively ensure the bacterization of the EO / EOMs, either directly by lamination of the granules already produced and consolidated, or indirectly, if possible, by incorporating said inoculum during their manufacturing processes.

  In summary, the invention therefore comprises (FIG. 1): É A first contact of the BFCPs with fine soil particles serving to boost their multiplication during said fermentation and solid state formulation (FFES), allowing the manufacturing a BFCP inocula, advantageously in the form of a more or less fine powder, with a sufficient cellular title. The preferred method for obtaining the candidate BFCPs which are particularly well suited to living in the soil is described in FR 2 833 016.

  É A second contact between the inoculum BFCP thus produced and the biosolids carried out either at the time of the filming of the EO / EOM with said inoculant powder produced by FFES, or by the incorporation of this inocula into a manufacturing process of EO / EOM granules having no ultimately bactericidal step with respect to the candidate BFCP hardy and / or isolated according to the protocol FR 2 833 016 (Claude 2001) and may therefore adversely affect and / or permanently to the survival of said BFCP .

  1st contacting: Manufacture of the inoculant powder by FFES According to one embodiment of the invention, a consortia of BFCP type cells, advantageously azotobacterial, given the ease of their culturing once isolated according to the protocol described in FR 2 833 016. These BFCP cells may, for example, come from Québec's arable soils such as sandy podzols and / or clayey gleysols. We can also highlight the usefulness of FFES using previously isolated azotobacterial strains according to FR 2 833 016 (Claude 2001). These bacterial strains, BFCP as it should be according to the claims of FR 2 833 016, are particularly well adapted to life in the soil, and therefore capable of interacting favorably with the fine soil particles forming an integral part of the FFES.

  It is not imperative to use a consortium of n candidate strains to start the FFES; everything depends on the nature of the soil, the in situ functioning of the strains. The FFES, by the presence of fine soil particles, also has the advantage of contributing itself to ensuring a certain selection of the most edaphically competent strains (ie adapted to life in arable soils). In addition to providing the production and formulation of bacterial biomasses of telluric origin, the FFES according to the present invention thus also makes it possible to ensure, to a certain extent, the edaphic competence. At the end of the FFES, the BFCP cellular titles of the n members of the initial consortium may not necessarily be identical. That said, for the purpose of registering a potential commercial product, the use of a single BFCP strain to start FFES is probably the most beneficial.

  The production and stabilization of these candidate bacterial BFCP cells on an organic support, for example residues of chopped cereal crops of approximate dimensions of the order of 1 to 4 mm, allow them to interact with both this support, which can also serve as a source of carbon substrate, and fine particles of arable soil, preferably about 100 micrometers, clay or any other inorganic and / or mineral support of a colloidal or clay character capable of interacting with humic substances or other phenolic polymers, natural or synthetic, characteristic of arable soils. The fine soil particles, according to the preferred embodiment of the invention, can be considered as analogous to a functional fraction of the clay-humic soil complex (Mills 2003). In order to allow this inorganic support x bacterial x organic support interaction, it is advantageous to proceed as follows: coating, by spraying, kneading, spraying or immersing, a portion of substrate / carbon supports, advantageously cereal crop residues including corn - grain, with about 1 e6 cells of each of the few BFCP strains retained as research component of the possible inoculum, per gram of substrate / support, preferably a residue of non-sterile cellulosic culture; - Again re-coating, advantageously by mixing, the culture residues with said fine soil particles so that the substrates / supports are completely coated with these particles. In principle, the effectiveness of this coating is recognizable by the absence of unadhered fine particles. It is advantageous to provide as many fine particles as organic support, although the proportions may vary depending on the metabolism of BFCP.

  Once the desired cellular titre has been achieved (from 109 to 1010 cells per gram), the mixture can be brought to a relative humidity level of about 18%, or even preferably about 12 to 15%, which is sufficiently dry to be pulverized by grinding in the form of an inoculant powder of variable granulometry according to need (see below, 21st contacting).

  In this sense, because of the ability of edaphic bacterial cells to interact favorably with the fine soil particles, or analogs thereof, coating the chopped crop residues, the invention is particularly well-suited for the formulation of nitrogenous biomasses and BFCP as described in FR 2 833 016. According to the preferred embodiment of the invention, the procedure is as follows: Sterilize, for example by autoclaving, the culture residues finely minced beforehand; - Sterilize, for example by autoclaving, the fine soil particles; Lo treatment with an antibiotic, ampicillin for example, an aqueous extract of a mixture of soil (fresh or pre-incubated) and residues of straw-rich crops (including those of corn), advantageously 50/50 w / w , in order to ensure a clear microbiological predominance of the fungal biomasses in these extracts (here, it is necessary to be very careful to sufficiently wash the solid fraction of this extract, for example by centrifugations and re-suspensions, in order to eliminate the antibiotic used); - Bring to culture residues, by equal volume by weight, 1 7 bacterial (azoto) cells per gram of residues; it is necessary here to use, as liquid medium of co-inoculation, said fungal-dominated extract prepared above; After mixing the culture residues thus inoculated, incorporate the fine particles of soil and knead, under aseptic conditions, preferably by stirring; Incubate, once the container is sealed, at least two to three weeks in order to ensure the development of cellular titre at a level equal to or greater than 5x109 bacterial cells per gram of solid support, or even more if allowed.

  This mixture of crop residues / fine soil particles / fungal extract and bacterial biomasses is now the raw inocula and solid support formulated in the solid state. It is equally advantageous, instead of sterilizing crop residues and fine soil particles, to subject them to thermal shock (for example, heating them to about 60 degrees Celsius for an hour in an oven) in order to mitigate any bacterial and fungal contaminants they may generate during incubation. This approach has the advantage of not overly harming the incubation dynamics, but still leaves an appreciable amount of contaminants, below the 1 6 germs per gram of inocula, the upper limit allowed by the legislation. course in France.

  In order to facilitate the filming of the EO / EOM, if necessary, it is advantageous to finely grind the inoculum thus produced and its solid support. o Simple mechanisms like coffee grinder or food processor at

  Rotative actions are useful in this sense. Due to the low relative humidity of the preparation, this milling and sieving (eg 750 micrometers) is easily achievable and does not cause the death of the bacteria.

  There is no technical limit to the size of each batch of production, since a solid state fermentation is feasible, for example according to the methods of Suryanarayan et al. (2001), Inra (1990), or according to the rules of art generally described in Raimbault (1998). It is also possible to make small batches, individually from 1 to 5 kg, or even 10 kg, final weight post-FFES, the equivalent of a hectare dose. Finally, the use of a solid state fermentation process in concert with said FFES as described herein can advantageously serve to reduce the production time, for example here and without limitation, well below the 21 days required to produce the desired cell title mentioned above.

  2nd Contacting: Incorporation of BFCP in EO / EOM The inocula, and more particularly the BFCP cells that it contains, produced during the first contacting can now be put in contact with the EO / EOM, or, where appropriate and if possible, the organic fraction of the premixtures to be used for their manufacture.

  Given a certain fragility inherent in all bacterial cells of telluric origin, it is advantageous that the EO / EOM and / or the aforementioned premixes contain a certain proportion of organic carbon, advantageously a minimum of 20 to 25%. That said, and given the hardiness and the edaphic competence of bacterial cells isolated according to FR 2 833 016, the mineral counterpart preferentially MAP (mono-ammoniacal phosphate salts) can reach the equivalent of 25% of the final composition, even more, for example 30%, given the presence of organic binders and / or according to the organic content of the constituent biosolids.

  Bacterization by contacting the inocula BFCP and granular EO / EOM and / or their organo-mineral precursors can be carried out either on the farm by contacting the inocula and said EO / EOM, or industrially by way of a conventional seed treatment method, and this insofar as said method is not per se bactericidal. In both cases, it is necessary to incorporate the equivalent of the hectare dose, advantageously from 1 to 5 kg of the cell titre inocula of at least 5e9 candidate BFCP cells per gram, which is equivalent to a hectare dose of approximately 1E13 to 5E13 via 200 kg of EO / EOM, or according to the needs of the crop.

  Regarding the bacterization of EO / EOM pellets postconstitution, and given the desired small size of inoculum particles and the roughness of EO / EOM granules, the adhesion of the inoculum is ensured without having to add water or any binder, although these may be useful in certain circumstances; this spontaneous adhesion, where appropriate, has the advantage of minimizing bactericidal physicochemical reactions near the granules. Finally, the EO / EOM are advantageously granulated, either in the form of cylindrical plugs or in spherical form, less common but more advantageous packaging (see EP 1 174 404 A2) With regard to the incorporation of BFCP by contacting with the premix during the manufacturing process of said EO / EOM, it is advantageous that this process does not include bactericidal steps, especially 3o by physicochemical and / or thermal treatments such as consolidation by baking at high temperature during extended periods of time.

  According to the various embodiments described in Tables 1 and 2, it is advantageous to include a binder, preferably organic, such as molasses and / or beet vinasse at a level of 5 to 8% (mass basis). In addition to allowing better consolidation of the granule, this type of binder can serve as a carbon substrate for BFCP cells. According to one embodiment, the granules can be gradually dried at a relatively low temperature, such as, for example, the room temperature of a room. From a bio-industrial point of view, however, it is sometimes advantageous to use a higher drying temperature in order to reduce the time required for this consolidation. This approach is also compatible with the present invention, given the hardiness of the BFCP (AZM) strains isolated according to FR 2 833 016.

  Thus, the incorporation of BFCPs can be carried out, either before or after the formation of the granules. If the manufacturing process involves a potentially bactericidal step, it is advantageous to incorporate the BFCPs after the formation of the consolidated granules. To do this, it is advantageous to modulate the porosity of the granules and the particle size of the inoculant powder so as to allow said inoculant to be embedded in the granules once they have been produced. If the manufacturing process does not include a potentially or effectively bactericidal step, it is advantageous to incorporate the inoculant powder produced by FFES directly to the premixes.

  Furthermore, the present invention also relates to a conditioning of plant growth promoting bacterial (BFCP) formulations as obtained prior to mixing with fertilizers, in the aqueous phase comprising: (a) a formulation of BFCP produced and formulated at the solid state contained in a first container, (b) an aqueous phase in a second container, the first and second container being arranged such that the microorganisms contained in said formulation are in suspension in the aqueous phase.

  Preferably: - said conditioning is characterized in that the formulation of BFCP in the solid state is immiscible with water.

  said conditioning is characterized in that the BFCP formulation is inorganic or organic.

  s - said conditioning is characterized in that the formulation contains fine soil particles, or their analogues.

  said conditioning is characterized in that the second container comprises a volume sufficient to allow said suspension of the micro-organisms, advantageously from 1.5 to 10 L. - said conditioning is characterized in that the second container is an infusible bag with inside the first container 1 closed and integral.

  said conditioning is characterized in that the second closed container consists of a suitable porous membrane enclosing said microbial formulation immiscible with water.

  said conditioning is characterized in that the first container is not closed and screwed on its lower part to the second container, containing said microbiological formulation immiscible with water.

  said conditioning is characterized in that the first container contains in its lower part a grid for supporting a porous membrane.

  said conditioning is such that the first container contains an orifice making it possible to adjust therein means for spraying the suspension.

  PREFERRED EMBODIMENT AND EXAMPLES OF APPLICATIONS From small samples of two soils (Table 1) were isolated, according to Claude (2001), pure cultures of AZM candidate strains, that is to say diazotrophic and capable of to solubilize inorganic recalcitrant phosphate compounds, ie a priori and otherwise poorly soluble in water. Aliquots of the resulting mother cultures had to be transferred to JM media (without nitrogen, Kumar and Narula 1999). The numerical dominance of these AZM cells will be provided by liquid JM cultures of Rang III. JM media contain as the sole source of phosphate di- and tricalcium phosphates and in the form of hydroxyapatite, or HA. Colonies will be obtained by plating aliquots of Tier III cultures on JM and PK agar media (with N, Kumar and Narula 1999) also containing insoluble di- and tricalcium phosphates. The formation of translucent halos around certain colonies will indicate the dissolution of these previously insoluble phosphate compounds.

  In order to evaluate the solubilizing capacity of the AZM isolates, the pH of the JM media was determined after seven days of culture for the (R) I, II and III cultures; the lower the pH, the greater the solubilizing power of AZM isolates, in principle (Kim et al 1997, Kim et al 2003). For practical reasons, only twelve colonies, six per soil type, were selected based on their solubilization indices (IS, Kumar and Narula 1999); these colonies will be used for the production of AZM biomasses in No.1 culture medium (Institut Pasteur, Paris) of AZM cell titers of approximately 1e9 per mL.

  Each of the 12 isolates (6 per soil) were evaluated individually against (12) non-bacterialized matched controls (t); each comparison is repeated three times. Dry matter production of aerial parts, or MSPA (Hilali et al 2001, Gachon 1988, Sardi and Csatho 2002) will serve as an index of the in situ activity of these candidate AZM isolates.

  The twelve isolates will be tested in soils A and S reconstituted (170 g / pot) and seeded with millet (Phleum pratense L.). Culture residues are used as expedients for AZM (1% w / w); the cultured residues thus contained 5E5 AZM cells per gram, or approximately the equivalent of the hectare dose reported by Claude and Fillion (2004) for the in situ bacterization of crop residues in agronomic situations. The pots, 72 in all, were fertilized with a nutrient solution (100 ml / pot) devoid of nitrogen and phosphorus, the only exogenous phosphorus introduced by solid incorporation being the equivalent of 60 kg of P per ha in the form of hydroxyapatite (HA). The equivalent of 75 units of nitrogen (ammonium nitrate) per hectare is provided 28 days after sowing, 10 to 12 days before the first cut (Cl). In order to evaluate the effect of the HA itself and / or culture residues, two controls were included; reconstituted soil without HA or crop residues, and soil reconstituted with HA, the controls with HA and culture residues being paired with each of the infected pots using one of the 12 candidate AZMs.

  Depending on the ability to favor the production of MSPA, it is possible to eliminate the few AZM candidate strains the least promising. The resulting AZM candidates will be used to initiate a solid state fermentation and formulation process, or FFES, to produce an inocula for the bacterization of EO / EOM precursor biosolids. The cell title the solid inocula thus produced can advantageously reach 5E9 AZM per gram.

  o Each successful AZM will be produced in this way, with the final consortium being an equal part of each one. The evaluation of the 12 AZM (T) isolates is performed using 12 paired non-bacterial controls (t), each experimental unit thus constituted being repeated three times. The T / t ratios for each isolate will serve as criteria for evaluating the in situ efficacy of the candidate AZMs. These strains were subsequently produced and formulated according to the fermentation and solid state formulation method according to the present invention.

  Table 1: Physico-chemical characterization of the soil series Batiscan (soil S sandy podzol) and Le Bras (clay soil Gleysol A) from which both soils originate; these two sols are also used for application examples 3 and 4.

  Soil Characteristics Soil S Series LeBras Batiscan Classification Gleysol Podzol Sandy Loam Texture Clay Soil Deschambault, Locality St. Lambert, Que. Qc% Sable 17 56 Soil 63 34% Clay 20 10 5.87 5.95 2, 61 3.26 11.6 14.1 45.4 76.4 45.1 154 898 712 80.9 37.5 972 1588 169 78 7, 5 15 4.68 4.81 62 31 o To better ensure the realization of the In the invention, it is advantageous to bacterize EO / EOM containing at least approximately 20% carbon, ie approximately 45 to 50% of organic matter. This approximate minimum content avoids too much physico-chemical reactivity of the granules in contact with the soil, reactivity contrary to the immediate survival of the BFCP, while providing them with a maximum of carbon substrate in addition to that exuded by the roots.

  Banding in the vicinity of the seeds as a starting fertilizer: EO / EOM should be applied near the seeds using a combined seed drill as a starter fertilizer (Craaq 2003). This is often the case with spring planting of field crops, including corn and some spring cereals (CPVQ 1988). Fortunately, these situations are also generally characterized by the absence of crop residues on the soil, or on the surface of the soil, at the time (spring) of the sowing and thus the placement of starter fertilizer in strips.

  ^ Soils lacking large amounts of crop residues at planting time: the presence of an overabundance of crop residues, which is more than a water pH M.O.

CEC

  mg / kg P mg / kg K mg / kg Ca mg / kg Mg mg / kg Al mg / kg Fe VEC (% P / AI) E / AI (P / AI) / VEC three or even 5 mg per hectare at planting time, will favor the strictly organo-mineral action of the bacterial EO / EOM at the expense of highlighting the relative efficiency of BFCP. More precisely, it is the relatively low C / N ratio (about 30) of the organic component of EO / EOM, relative to that of crop residues (about 100), which will allow this organic material to initiate some mineralization of these residues. It is therefore advantageous to jointly apply BFCP and EO / EOM in situations where crop residues on the soil are little or not present at the seedlings.

  In the case of BFCP intended for the improvement of the phosphate nutrition of the cultures, it is advantageous that the dominant speciation of the most present phosphatic compounds is in agreement with the nature of the MPS power of the BFCP concerned. More particularly, soils that are rather rich, or even saturated with alumino-ferric phosphate compounds, for example podzols, must receive isolated BFCPs according to their MPS powers directed towards this type of compounds. Alternatively, soils that are rather rich, or even saturated with calcio-magnesium phosphate compounds, should rather receive isolated BFCPs according to their MPS capacities directed towards calcium phosphates, for example hydroxyapatites.

  The utility of the invention is not limited to the bacterization of the EOMs since the EOs too can, in principle, be applied in strips near the seeds there and where the application of EO / EOM would not be possible. possible when planting winter cereal crops in France, for example.

  Although the practice is not common, it could become so through the advantages of the invention. In fact therefore, and to facilitate the description of the present invention, the concept of EO / EOM does not exclude that of EO.

  Compared to a more conventional solid-state fermentation, ie without the addition of fine soil particles capable of interacting favorably with bacterial strains of telluric origin and particularly well adapted to life in soils, we can therefore significantly increase the cellular titer of these preparations (Figures 2 and 3). Since these preparations are rather dry (ie 15% humidity), they can be considered as formulated in the solid state and can be used, inter alia, for the bacterization of EO / EOM, or even their premixes in the course of extrusion and consolidation. If it were not for this increase in cellular titre, the costs normally associated with the preparation of such an inoculum composed of rustic and generally oligotrophic telluric bacterial strains would be prohibitive and the bacterization of EO / EOM would be too expensive.

EXAMPLES

  Example 1: Effect of the coating of fine soil particles on the nitrogen cell density of a mixture of crop residues (wheat) and fine soil particles (70 microns) on the accumulation of ammonia nitrogen and Azotobacterial biomasses over time.

  In the context of an FFES according to the present invention, we determined the azotobacterial cell density of the AZM strains with and without the presence of fine soil particles (Gleysolic clay loam). The presence of fine particles of soil (Figure 2) effectively makes it possible to increase the density of the azotobacterial cell hundredfold. Only the AZM azotobacterial strains isolated according to FR 2 833 016 (Claude 2001), or even, to a lesser extent, with the control nitrogenous strains isolated according to FR 2 833 016, are capable of interacting favorably in this way with said fine soil particles. and achieve an important cellular title; the cellular titre obtained with a control azotobacterial strain (Azotobacter, chroococcum No. 76.116, Collection of the Institut Pasteur, Paris) is it, much lower.

  Example 2 Evolution of Cellular Densities Within Microcosms Containing the Bacterilized Culture Residues and Coated with Fine Particles of Soils According to the Invention

  Most of the protocol used for Example 1 was taken up again but only with the microcosms containing the fine soil particles in order to better demonstrate the kinetics of accumulation of the different bacterial biomasses according to their origin and the type of isolation protocol used. .

  As shown in FIG. 3, as of the 7th day of incubation, only the FFES containing the fine particles of soil inoculated with AZM azotobacterial biomasses isolated according to FR 2 833 016 (Claude 2001) contain more than 1 e8 AZM azotobacterial cells per gram of preparation. This clearly demonstrates the possibility of simultaneously producing and packaging nitrogen-containing inocula, and that if and only if, the bacterial biomasses concerned are particularly well suited to life in soils and therefore all the more capable of interacting with these fines. soil particles, which is not the case, for example, of the A. pleuro azoclobular A. crroococcum 76.116 biomass of the Institut Pasteur, or even of the other control biomasses produced according to FR 2 833 016. In the case of candidate azotobacterial strains isolated according to FR 2 833,016, the cell density increased from 1e6 to 1s5e9 after 21 days of incubation; the control strain sensu FR 2 833 016 was, as expected, more prolific than Azotobacter 76.116, without reaching the cellular titres achieved with AZM.

  Table 2: Physico-chemical characterization of the 9 biosolids (BS) / MAP / AZM premixes, including a calculation of the potential contribution of MAP to osmolarity (osmotic stress, N-MAP / N-Total and P-MAP / P- Total).

  Map origin M% N% P% K% M P / N PIK% NI% P I N% P biosolid Map org. MAP MAP MAP / MAP / N total sum GSI - SLPt 0.0 0.00 3.70 3.90 0. 52 64 1.05 7.50 37 20 0 0 GSI - SLPe 0.0 0.00 3.40 3.80 0.48 54 1.12 7.92 34 19 0 0 Shock BM S.0251 0.0 0.00 1.00 2.08 1.60 81 2.08 1.31 10 10 0 0 Shock BM s.0281 0.0 0.00 1.27 1.49 1.27 73 1.17 1.18 13 7 0 0 GSI - SLPt 4. 0 0.28 3.95 4.55 0.50 61 1.15 9.11 40 23 6 14 GSI - SLPe 4.0 0.28 3.66 4 45 0.46 52 1.21 9.66 37 22 7 15 Shoc BM S.0251 4.0 0.28 1.36 2.80 1.53 78 2.06 1.83 14 14 27 26 Shock BM s.0281 4.0 0.28 1.62 2.23 1.22 70 1.38 1. 84 16 11 22 33 GSI - SLPt 7.7 0.58 4.18 5.14 0.48 59 1.23 10.70 42 26 12 24 GSI - SLPe 7.7 0.58 3.91 5.05 0.44 50 1.29 11.39 39 25 13 25 Shock BM 7. 7 0.58 1.69 3.46 1.47 75 2.05 2.35 17 17 41 40 S.0311 Shock BM 7.7 0.58 1 94 2.92 1.17 68 1.50 2.49 19 15 35 49 S.0341 GSI - SLPt 14.3 1.26 4.60 6. 20 0.45 55 1.35 13.91 46 31 20 37 GSI - SLPe 14.3 1.26 4.34 6.11 0.41 46 1. 41 14.86 43 31 22 38 a Shoc BM 14.3 1.26 2.29 4.64 1.37 69 2.03 3.40 23 23 56 55 S.0311 Shoc BM 14.3 1.26 2.52 4.14 1.09 63 1.64 3.81 25 21 50 64 S.0341 GSI - SLPt 25.0 2.88 5.28 7.93 0.39 48 1.50 20.32 53 40 30 51 GSI -SLPe 25.0 2.88 5.05 7.85 0.36 41 1.55 21.81 51 39 33 52 Shock BM 25.0 2. 88 3.25 6.56 1.20 61 2.02 5.49 33 33 69 68 S.0251 Shoc BM s.0281 25.0 2. 88 3.45 6.12 0.95 55 1.77 6.44 35 31 63 76 Table 3: Physicochemical characterization of the six granular preparations acting as EOMI prototypes according to their levels in organic matter, dry matter, organic binder and N, P and K nutrients. The relative content of P and K directly from MAP as an osmolar index (osmotic stress) is also indicated.

  Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 70 70 70 70 70 Shoc Shoc Shoc GSI GSI GSI 25.1 28.1 28.1 Peat Peat (t) (t) 81 73 73 64 54 54 5 8 8 8 12 25 25 25 25 30 100 100 100 100 100 0 0 0 0 0 0 50 50 50 50 50 100 100 100 100 100 240 250 253 253 253 262 137.5 137. 5 137.5 137.5 137.5 120.0 112.5 115.5 115.5 115.5 124.5 29.2 28.0 27.7 27. 7 27.7 26.7 2.1 2.0 3.2 3.2 3.2 4.6 10.4 10.0 9.9 9.9 9.9 11.5 8.3 5.0 4. 9 4.9 4.9 4.8 33.0 26.4 26.7 24.2 21.5 21.5 50.0 45.0 45.7 45.7 45.7 47. 5 58.3 62.2 60.6 60.6 60.6 56.2 4.2 4.4 6.9 6.9 6.9 9.6 20.8 22.2 21.6 21. 6 21.6 24.1 g BS type BS% MO of BS g inocula BFCP g MAP mlH2O ml binder (50% water) ml binder (25% water) MO dry binder Total weight (g) ml H2O total g MS total É BS ÉFFES É MAP 0/0 Binder É MO 0/0 MS É BS / MS ÉFFES / MS É MAP / MS% Binder / MS 16.7 11.1 10.8 10.8 10.8 10.0 cYo MO / MS 66.0 58.8 58.5 53.1 47.0 45.2 of which% of BS 72 77 76 73 70 67 N in BS / MS 0. 0100 0.0127 0.0127 0.0370 0.0340 0.0340 P in BS / MS 0.0 208 0.0149 0.0149 0.0390 0. 0380 0.0380 K in BS / MS 0.0160 0.0127 0.0127 0.0052 0.0048 0. 0048 N 2.67 3.01 2.93 4.41 4.23 4.32% P 5.38 5.37 5.23 6.69 6.63 6.96 K 0.93 0.79 0. 77 0.32 0.29 0.27 N / MAP 26.7 30.1 29.3 44.1 42.3 43.2 P / MAP 26.9 26.9 26.2 33.5 33.2 34.8 N-MAP / N 78.1 73.8 73.8 49.1 51.2 55.8 P-MAP / P 77. 4 82.7 82.7 64.7 65.3 69.3 gMO / gN-MAP 31.68 26.44 27.04 24.52 21.72 18. 77 gMO / gP-MAP 15.84 13.22 13.52 12.26 10.86 9.38 gMO / gN-total 24.75 19.50 19.95 12.04 11.13 10.46 gMO / gP-total 12.26 10.94 11.18 7.93 7.09 6.50 Example 3: Resistance and survival of selected AZM-type BFCP cells according to the protocol of Isolation described in FR 2 833 016 and subjected to osmotic stress attributable to high doses of MAP used in the composition of said IMO: A third example of application made it possible to demonstrate the extraordinary resistance of AZM strains to osmotic stress caused by relatively high doses of MAP in the body of the EOMI. To do this, premixes of various types of biosolids and various doses of MAP were incubated; certain physico-chemical characteristics of these premixes are presented in Table 2. The hardiness of the AZM cells from FR 2 833 016 (Claude 2001) can be advantageously valorised during the second contact with a high concentration of mineral fertilizing salts, here about 25% of MAP.

  The presence of these salts is useful, even necessary given the low content of fertilizing mineral salts of EOM compared to that of MAP. Nevertheless, the content of mineral fertilizing salts of the possible EOMI will be necessarily and appreciably less than that of their conventional counterparts devoid of organic matter. This impoverishment, far from being a disadvantage, represents an agro-environmental advantage in that it makes it possible to reduce the dose-hectare of phosphorus introduced at start-up; it is the BFCP, advantageously of the AZM type, which will have to compensate for this beneficial phosphorus deficit.

  The ability of AZM cells isolated according to FR 2 833 016 to resist osmotic (osmolar) pressures caused by MAP concentrations up to 3 M MAP has been evaluated. 25% MAP on a mass basis (Tables 2 and 3). These concentrations are several times higher than those present in the soil solution and therefore a priori and in principle bactericidal except for the inherent hardiness of the AZM isolated according to FR 2 833 016. To highlight this greater hardiness, Control strains, called azm, diazotrophs and conventionally isolated MPS target soils (Table 1), were used. Obtaining this type of control strain was previously described in FR 2 833 016, and repeated above in the context of the present patent application. One e8 g of AZM and AZM strains were introduced into two series of biosolids / MAP mixtures more or less enriched in MAP (Tables 2 and 3) and previously sterilized by autoclaving; the cellular titre of these media was subsequently determined on PK and JM media (Kumar and Narula 1999, Table 3). These two strains were introduced into the same mixtures, but this time not sterilized, to evaluate their respective capacities to dominate all the microbial flora present (Table 4).

  Table 4: Cellular titre (mean for the nine (9) BS tested) of the azm control cells and the rustic AZM cells according to the monoammoniacal phosphorus salt (MAP) concentration of the biosolids premixes / MAP / BFCP (AZM, azm). The iMPS index, or dt28-3, is evaluated according to the innovative analytical method of the invention.

  Osmolarity concentration cells x e9 / g AZM / azm iMPS (dt28-3) MAP (% w / w) MAP (osM) azm AZM 0 0 10 10 1 -0.300 4 0.26 4 9 2 -0.175 7.7 0.58 1 8 8 -0.050 14.3 1.26 0.1 5 50 0.120 2.8 0.033 2 61 0.070 It is important to note that it is impossible to date to discern with certainty between these two strains, or between them and the other Io BFCP MPS soil, using biomolecular and genomic tools since these bacterial biomasses are related, apart from their hardiness. Indeed, and apart from their actions in situ on the growth of plants, the edaphically competent strains from FR 2 833 016 are difficult to identify and / or distinguishable from related strains that are less so.

  The osmotic stress provoked by the MAP is useful here in that it makes it possible to highlight this greater competence. In fact, the osmotic pressure ensures the dominance of the most rustic strains, and more particularly here the AZM, reintroduced into the soil according to the present invention. This dominance can be illustrated using an analytical method developed in the context of the present invention. It is a question of confronting the introduced strains with multiple stresses and to make the diazotrophy and the MPS character of these strains of the traits rapporteurs for the supposed hardiness of these strains. The inventors have developed an analytical method making it possible to ascertain the effective dominance of the AZM strains, the most hardy ones according to FR 2 833 016 (Claude 2001), at the expense of those which are less so. This method is thus advantageous for quality assurance and quality control (AQCQ) without having to depend on a fine genetic and / or biomolecular characterization of the edaphic competence. The inventors have therefore developed in this sense the following analytical method, called MPS or iMPS index, or dt28-3; Aliquots of a dilution of the premix during incubation are placed in a liquid medium of the JM type (Kumar and Narula 1999) free of nitrogen and soluble phosphorus; the only source of phosphorus being hydroxyapatite (HA), a recalcitrant and insoluble phosphate compound, except for the MPS action of the AZM and azm strains. The aliquots are diluted in this manner at pH = 17, and pH = 9 and the JM tubes incubated for 28 days at room temperature (20-22 degrees C). The optical density (OD) of the JM media is determined after 3 and 28 days of incubation, the difference between the two observations is indicative of the MPS activity attributable to the AZM or azm strains. In principle, if the aliquot consists only of diazotrophic and MPS strains, the difference t28-t3 will be large; the presence of non-diazotrophic contaminant strains and / or MPS will reduce the size of this difference. Therefore, and given the supposed hardiness of the AZM strains from FR 2 833 016 compared to the control azm cells, the aliquots from the high MAP premixes should contain a higher proportion of AZM, the other constituents of the flora. bacterial, including azm inoculated with AZM, having been removed by bactericidal osmotic pressure; the difference t28-3 will therefore be rather large if and only if the AZMs (MPS here) have actually resisted the osmotic stress. If the MAP content of the premixes is reduced, the selection in favor of the AZMs and at the expense of the other less hardy bacterial cells, and for the most part non-diazotrophic and / or MPS, will be less strong and the development of the optical density from t3 to t28 will be even less pronounced. In this sense, the MAP induces a selective stress in favor of the rustic BFCP (AZM) strains isolated according to FR 2 833 016, whereas the JM medium promotes the expression of the MPS character of these diazotrophic strains. The character MPS, and incidentally the development of the DO from 3 to 28 days post-inoculation of JM media, is here a reporter trait of the hardiness of AZM.

  In other words, the dissolution of phosphorus as detected by the initial clarification of the JM media, acts here as a reporter trait for the cells capable of diazotrophy, diazotrophy which is, by definition, BFCP-AZM, associated with hardiness supposed of these cells. If, in response to osmotic stress, the non-hardy cells are removed, only the rustic (AZM) cells can be subjected to a deficiency of nitrogen and phosphorus characteristic of the JM media, and contribute to densify (ie increase the apparent density) of these environments. To verify that it is the most hardy cells (AZM) reintroduced by inoculation that resist, and not other types of cells, in principle, the hardy months, even the control cells (azm), also reintroduced by inoculation, it s' is simply to include uninoculated controls and controls inoculated with control cells as defined in patent FR 2 833 016 (Claude 2001). Insofar as the uninoculated and inoculated (azm) controls do not cause a significant increase in the bulk density of the JM media as a function of the increase in osmotic stress in premixes with MAP, the densification of the associated JM media premixes inoculated with the so-called hardy BFCPs can only be attributed to the resistance of said cells to said osmotic stress. The increase in the optical density (densification) of the JM media, or even here of the iMPS as defined above, is therefore necessarily a positive demonstration of the hardiness of the candidate strains (AZM) introduced by inoculation into the biosolids premixes. / MAP, potential precursors of EOMIs. Better still, this method of analysis also makes it possible to evaluate, not only different candidate strains for their degrees of hardiness, but also different matrix substrates for their bactericidal levels (see Tables 5 and 6, and Figure 4). As demonstrated in the context of the present invention, this method of analysis is particularly suitable and useful for the screening of candidate strains and prototype biosolids which can serve as precursors to the manufacture of EOMI. This method is particularly useful for the evaluation of resistance to various stresses whose genomic and metabolic mechanisms are (necessarily) complex and thus little or no elucidated (elucidable) and for which, therefore, no biomolecular or even bioinformatic method has has been developed to date.

  Table 5: Estimate of AZM dominance according to the dt28-3 index (or iMPS) according to the various types of biosolids (BS) evaluated and the MAP concentration of biosolids premixes / MAP / BFCP (AZM, azm); these data are reported graphically in Figure 4.

  Biosolids (BS) 0% MAP 4% MAP 8% MAP 15% MAP 25% MAP iMPS (dt28-3) Envirogain (BS 1) Lisox (BS 2) Lisox C (BS 3) Powder (BS 4) GSI Centri (BS 5) -0.429 -0.161 -0.127 0.052 -0.015 GSI-SLPt (BS 6) -0.141 0.056 -0.047 0.189 0.315 GSI-SLPe (BS 7) -0.388 -0.096 -0.151 0.122 0. 139 Shock 25.1 (BS 8) -0.326 -0.105 -0.092 0.063 0.155 Shock 28.1 (BS 9) -0. 445 -0.256 -0.225 0.091 0.018 Average -0.346 -0.135 -0.128 0.103 0.122 This analytical method also has the advantage of highlighting the effect of the various biosolids on the selection to the advantage of AZM (hardy) 20 and the costs azm (non-hardy) thus favoring a dominance of the AZM in the biosolids / MAP mixtures and possibly also in the vicinity of the EOMI granules in situ. According to our analysis, the iMPS index (or dt28-3) increases steadily according to the osmotic stress resulting from the increase in salt content directly from the MAP (Tables 5 and 6). This increase in iMPS is all the more detectable as the organic matter (OM) content of the biosolids is low (see Tables 2 and 3); the low OM content allowing free MAP salts to act on the survival of non-hardy cells to the advantage of AZMs that are. According to this analysis, reported in Figure 4, it is possible to clearly distinguish between the biosolids rather low in MO, ie here the biosolids GSI (see BS 5, 6 and 7), and those which are rather rich in MO, ie to here the so-called Shoc biosolids (see BS 9 and 8). This indirect but appropriate measure of bactericidal biosolids / MAP mixtures serves to intelligently influence the choice of biosolids to be used for the eventual constitution of prototype EOMIs.

  Table 6: AZM-azm cellular titers and AZM iMPS, or dt28-3, dominance indices for various EOMI granular preparations according to their MAP content and their N and P concentration relative to that of MAP; two modes of drying and consolidation of the granules have been tried.

  iMPS (dt28-3) Preparation (21C - 56h)% MAP% P / MAP azm AZM AZM / azm dt28-t3 BS 8 (Shoc 25.1) 20.8 27 0.1 20 200 0.155 BS 9 (Shoc 28.1) 21.2 27 0.08 50 625 0.018 BS 9 (Shoc 28.1) 21.6 26 0.06 40 667 0.018 BS 7 (GSI SLP bark) 21.6 34 0.04 44 1100 0.315 BS 6 (GSI SLP peat) 21.6 33 0.04 43 1075 0.139 BS 6 (GSI SLP peat) 24.1 35 0.05 45 900 0.139 Preparation (60C - 1h)% MAP% P / MAP iMPS (dt28-3) AZM / azm dt28-t3 azm AZM BS 8 (Shoc 25.1) 20.8 27 0.07 19 271 0.210 BS 9 (Shoc 28.1) 21.2 27 0. 05 45 900 0.026 BS 9 (Shoc 28.1) 21.6 26 0.05 39 780 0.021 BS 7 (GSI SLP bark) 21.6 34 0.03 38 1267 0.363 BS 6 (GSI SLP peat) 21.6 33 0.03 40 1333 0.173 BS 6 (GSI SLP peat) 24.1 35 0.04 39 975 0.151 In a first step, a range of biosolids / MAP / BFCP mixtures (here AZM or azm, Table 5) were evaluated for their degree of bactericide according to the index dt28-3 (or iMPS). As expected, the cellular titre is inversely proportional to the MAP concentration. Better still, AZM cellular titers are much more resistant to these osmolar stresses than azm cellular titers (Tables 4 and 5). At high MAP levels, AZM cellular titers largely exceed those of azm cells by a factor of more than 50, with azm probably less resistant to this type of stress. In addition, the ratio of cellular titers for AZM and azm cells is in perfect agreement with the change in the optical density of the JM media as determined by the iMPS index (dt28-3); this relative measure of the in situ dominance of the AZM type strains resulting from FR 2 833 016 thus reflects the hardiness inherent in these cells specifically selected for their edaphic competence and their adaptation to life in arable soils, including in proximity to granules of EOMI relatively rich in MAP salts. This analysis also distinguished between two groups of biosolids (Table 5 and Figure 4).

  Despite the addition of organic binders and the organo-mineral support necessary for the production and formulation by FFES of the AZMs, the bacterial titre, and thus the resistance to osmotic stress attributable to the presence of MAP, could probably be maintained due to the relative hardiness of AZM compared to less competent azm and adapted to life in soils (Table 6).

  Finally, the effect of a drying temperature higher than room temperature on the survival of post-extrusion granules was also briefly evaluated. The accelerated consolidation of these granules with a drying temperature of 65 degrees Celsius for a period of 90 minutes, sufficient to ensure the consolidation of the granules, had no marked and / or counterproductive effect. on the survival of AZM bacteria. According to the data reported in Table 6, AZM cells, and azm to a lesser extent, withstand such thermal shock. This hardness of AZM is perfectly in accordance with the present invention.

  Once the AZM strains produced economically by FFES, and their proven hardiness, especially as regards their resistance to osmotic stress near the EOMI granules, their agronomic utility has been demonstrated. To do this, azo-bacterial strains of the AZM type were re-isolated according to the protocol described in FR 2 833 016 (Claude 2001) and intended to be used in the manufacture of EOMI (see Examples 1, 2 and 3). Non-bacterial EOM homologs of these EOMIs (EOMs) have also been produced to clearly demonstrate the AZM effect on the relative weight efficiency of EOMIs. For the purpose of the experiment, and here by way of non-limiting example, there is the BS6 granular preparation containing 24.1% MAP (see Tests 6 in Table 2); the bactericide of this preparation is reported in Table 5. To produce the EOMI, 9.6% of the inocula AZM produced according to our FFES process was therefore incorporated on a mass basis. For the constitution of the non-bacterized EOM controls, 9.6% of the sterile co-formulating substrate was incorporated on a mass basis without the BFCP-AZM. This supply of EOMI makes it possible to send to the soil, close to the corn seeds, the equivalent of 13 cells AZM per hectare. The granulation of the premixes may be carried out by extrusion according to a recognized industrial process. As a conventional control, salts of MAP (11-52-0) were used as mineral fertilizer. A phosphorus-free control was also used. These EOMIs were tested, along with their controls in the greenhouse (Tables 7 and 8) and fields (Tables 9 and 10).

  The choice of soils and experimental sites has been the subject of particular attention. In soil relatively rich in P, MAP as a control will not be effective, and therefore the relative ineffectiveness of non-bacterial EOM can not be inferred; a possible effectiveness of the EOMI can only be attributed to the BFCP, which is good, but is not inventive in itself and / or the object of the present experimental development. In relatively poor soil at P, the action of the MAP will be detectable, and, although that of the EOMI a little less than in soil rich in P, the effectiveness of the EOMI compared to those of the EOM will be now more obvious, even comparable to MAP. In this sense, we have not simply sought to highlight the usefulness of shipping AZM-type BFCPs near the roots, which is not in itself inventive, but to do so with the help of EOMI, thereby improving the relative efficiency of an existing technology, EOMs. It is best to ensure that the MAP effect is detectable. In addition, and from a commercial point of view, it is interesting to demonstrate that the bacterization of EOM is effective on soil poor in P, or comparable to MAP used as a starter fertilizer. So, if the risk associated with this type of innovative o fertilization in a situation where the use of MAP is imperative, the use of an organomineralized starter is possible. In addition, the use of this new type of starter can contribute, through its BFCP component, to increase the yield potential on rich soils and phosphorus.

  Example 4 Efficiency of the EO / EOM Bacterisized versus Uninfected EO / EOM.

  As part of this greenhouse trial, two large groups of soils were used (Table 6); (1) Podzolic soils of the uplands in Quebec, whose phosphates are mainly in aluminoferric form (Fe, Al), and (2) Gleysolic soils of the Montreal plain (also known as the St. Lawrence Lowlands), including phosphates. are mainly fixed in calcio-magnesic form (Ca, Mg) (Tabi et al., 1990). For each of these large groups, two levels of phosphorus fertility were chosen.

  For each of these four (4) soil types, the selected prototype EOMI, its non-bacteriated counterpart (EOM), and a conventional starter fertilizer (MAP) were evaluated, as well as a control modality without starter fertilizer. These four (4) modalities were repeated four times and were arranged in Latin squares in two-liter pots containing 1.667 kg of reconstituted soils for greenhouse experimentation. The fertilizers were initially placed on 1,000 kg and covered with a layer of 333 g of soil; two maize seeds were placed at the top of these 1.333 kg of soil and covered with 333 g of soil. In this way, the fertilizers are placed to simulate the application of starter fertilizer in strips to the fields at the time of corn planting. In all, 64 two-liter pots were installed, monitored and harvested. The test plant used is maize harvested at 30 cm; the plants were irrigated with a phosphorus-free nutrient solution, but rich enough to ensure the full growth of seedlings according to the Craaq guidelines (2003). The dry matter of the aerial parts (MSPA) were determined and analyzed for their content of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg).

  To determine whether increases in MSPA yields, and possibly, if necessary, N, P, K, Ca and Mg samples attributable to EOMIs are significant, the four sets of 16 pots were arranged in 4x4 Latin squares. With the help of a conventional Anova analysis protocol, which was adapted to this type of experimental device, it was possible to compare the averages of the four modalities according to a Newman-Keuls (NK) technique.

  On soils with a P / Al saturation degree of P, i.e. (i.e., 5.0), the effectiveness of MAP as a starter fertilizer is undeniable; that of the EOM is much less so (Tables 7 and 8), mainly because of their lower P content and, to a lesser extent, N. This reduction in P contents, although desirable from one point from an agro-environmental point of view, is agronomically significant. The bacterization of these same EOMs, however, made it possible to achieve MSPA yields comparable to those obtained with MAP (Table 7 and 8), and this while achieving at the start of the crop a saving of about 26 kg of P2O5 and 5 kg of nitrogen per hectare. These results were reproduced on both types of soils. That said, if the degree of saturation of the soils is more appreciable (ie P / Al = 7.5), the effectiveness of the MAP, and incidentally that of the non-bacterized EOM control, is essentially zero, whereas the efficiency of the EOMI remains detectable; it is even more effective in MAP compared to less saturated soils in P. The action of the BFCP-AZM component of EOMIs is therefore independent of their organo-mineral components. In this sense, it adds to these and provides, under certain circumstances, tripartite action organic, mineral and bacterial EOMI.

  Table 7: Production of aerial matter dry matter (MSPA) on podzolic soil by maize plants grown in a greenhouse and harvested when they reached 30 cm in height depending on the starting mode.

  Podzolic soil Inputs Yields MSPA at start-up (maize 30 cm) Starting method N P205 K20 P / Al = 5.0 P / Al = 7.5 (kg / ha) (mg / pot) Indicator without starter 0 0 0 300 c 402 b With MAP (11 52 - 0) 9 40 0 425 to 426 b With EOM (4.5 - 17 0.5) 3.8 14 0.3 325 bc 412 b With EOMI (4.5 17 3.8 14 0.3 400 to 468 ab 0.5) Table 8: Dry matter production of aerial parts (MSPA) on a Gleysolic soil by maize plants grown in a greenhouse and harvested when they reached 30 cm in height according to the start mode.

  Gleysolic soil Contributions MSPA yields at start (maize 30 cm) Starting method N P205 K20 P / Al = 5.0 P / Al = 7.5 (kg / ha) (mg / pot) Indicator without starter 0 0 0 328 c 448 b With MAP (11 52 - 0) 9 40 0 450 to 455 b With EOM (4.5 - 17 0.5) 3.8 14 0.3 318 b 455 b With EOMI (4.5 17 3.8 14 0.3 436 to 492 ab 0.5) Example 5: Effectiveness of the EO / EOM bactericidal compared to EO / EOM uncharacterized.

  The procedure is the same as that used for Example 4. We used two large groups of soils (Table 6) in order to locate two contrasting experimental devices in terms of texture and pedology but whose saturation levels P (ie P / AI) are comparable. To determine whether increases in MSPA yield and N, P, K, Ca and Mg samples attributable to EOMIs are significant, we arranged the two sets of 12 plots (ie, four (4) ox oxo-repeat modalities) in blocks. Fischer (randomized complete block design), 4x3. Using a conventional Anova analysis protocol for variance adapted to this type of experimental device, we were able to compare the averages of the four modalities according to a Newman-Keuls (NK) technique.

  As expected, and given an average degree of saturation at P (ie P / Al of approximately 4.8), the MAP efficiency at both sites is detectable, whereas that of the non-bacterial EOMs is detectable. is hardly (Tables 9 and 10). The relative efficacy of the IMO is comparable to that of the MAP at the Saint Lambert Gleysolic site, and similarly to a lesser extent at the Deschambeault Podzolic site, but above all, significantly greater than that of the non-bactericidal EOMs. It should be noted that only the MAP treatment is supplemented with 20 units (kg / ha) of ammonium nitrate. This supplement is not added to the EOMI treatments for bactericidal reasons. Compared to MAP treatments (with ammonium nitrate supplement), the weight efficiency of the EOMI treatments is therefore superior.

  Table 9: Maize grain yields and nutrient samples (N, P, K, Ca, Mg) for a non-starter control on a Batiscan podzol (Deschambault, Quebec) according to different starting conditions P / Al = 4 8.

  Table 10: Corn grain yield and nutrient harvest (N, P, K, Ca, Mg) relative to a control without a starter on a podzol of the Le Bras series (Saint Lambert, Quebec) according to different start-up modalities P / AI = 4.7.

  Inputs at start-up Yield of grain corn / control without starter N P205 K2O grain NPK Ca Mg Starting method (kg / ha) (kg / ha) Indicator without starter 0 0 0 (1.00) (1.00) (1.00) (1.00) (1.00) (1.00) With MAP (11 52 - 0) 29 40 0 1.10 to 1.10 to 1.10 to 1.10 to 1.10 to 1.10 a With EOM (4.5 - 17.0 , 5) 3.8 14 0.3 1.03 b 1.02 b 1.02 b 1.01 b 1, 02 b 1.01 b With EOMI (4.5 17 0.5) 3.8 14 0.3 1.09 to 1.09 to 1.10 to 1.07 to 1.08 to 1.08 a Start-up efficiencies N P205 K2O (kg / ha) Yield of grain corn / control without starter grain NPK Ca Mg (kg / ha) Start mode Indicator without starter With MAP (11 52 - 0) With EOM (4.5 - 17 0.5) With EOMI (4.5 17 0.5) 0 0 0 (1.00) (1.00) ( 1.00) (1.00) (1.00) (1.00) 29 40 0 1.10 to 1.10 to 1.10 to 1.10 to 1.10 to 1.10 to 3. 8 14 0.3 1.02 b 1.01 b 1.02 b 1.01 b 1.01 b 1.01 b 3.8 14 0.3 1.06 ab 1, 06 ab 1.06 ab 1.06 ab 1.05 ab 1.06 ab References Aertsen , A., K. Vanoirbeek, Ph. De Spiegeleer, J. Sermon, K. Hauben, A.Far Ewell, T. Nystrom and C.W. Michiels. 2004. Heat shock protein-mediated resistance to high hydrostatitic pressure. Appl. About.

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Claims (27)

  A method of making organic fertilizer and / or organomineral fertilizer (EO / EOM) bacteriated by rustic plant growth promoting bacteria (BFCP) comprising the steps of: preparing a BFCP inocula; ii. fermentation and formulation of said inocula in the solid state (FFES); iii. contacting the powder obtained in ii with the EO / EOM.   2. The method of claim 1 as in step i. BFCPs are obtained from bacterial biomasses of telluric origin.   3. Method according to claim 1 or 2 such that the inocula of BFCP is obtained by a process characterized in that it comprises the steps of: placing an aqueous suspension of soil in contact with a matrix substrate, so as to to form a hydrated zone, or biofilm, on the surface of the substrate, analogous to that associated with the CAH of the soil, essentially sheltering the indigenous bacterial component of the soil, in order to retain on the surface of the matrix substrate, most of the endogenous bacterial microflora in the original edaphic media; - maturation of the biofilm with establishment of oligotrophic conditions allowing the most constitutive cells of the edaphic biofilm which colonized the matrix substrate, to migrate to the supernatant liquid phase the matrix substrate, and gradually to dominate it; - Recovery and culture of the most prolific bacterial strains in rich liquid culture environments, native, rustic and persistent in situ, cultivable in liquid media, and therefore able to be produced industrially.   4. Method according to any one of the preceding claims, such that the BFCPs are chosen from the families Azotobacteracea and Pseudonomacea.   5. A method according to any one of the preceding claims, such that the bacteria are diazotrophic and capable of dissolving the inorganic phosphorus attached to the soil particles.   6. Method according to any one of the preceding claims, such that the FFES of step ii. comprises contacting the BFCP with a substrate / carbon support and soil particles.   7. The method of claim 6 such that the carbon substrate / support comprises residues of cereal crops.   8. The process of claim 6 wherein the soil particles are from arable soils.   9. The method of claim 7 or 8 such that the soil particles are from analogs of the functional fraction of the soil argilohumic complex.   10. A process according to any one of the preceding claims, such that the moisture content of the powder obtained at the end of ii is between 10 and 25%.   11. Method according to any one of the preceding claims, such that the powder obtained at the end of step ii has a titre greater than 5x109 / gram.   The method of any of the preceding claims, such as contacting iii. is carried out by filming EO / EOM with the powder obtained in ii or by incorporating the powder obtained in ii into EO / EOM granules.   13. Method according to any one of the preceding claims, such that the composition obtained at the end of step iii has a cellular titre greater than 5x107 cells / g.   14. Process according to any one of the preceding claims, such that it does not comprise a bactericidal step.   15. The method of any one of the preceding claims, wherein the EO / EOM comprises at least about 20% carbon.   16. Process according to any one of the preceding claims, such that the EO / EOM are chosen from organic and organo-mineral fertilizers comprising biosolids, biomasses or other residual fertilizing materials.   17. Bactericidal organic or organo-mineral fertilizer (EO / EOM) comprising fermented plant growth promoting bacteria (BFCP) formulated in the solid state obtainable by the method of any one of claims 1 to 16.   18. Bacterialized organic or organo-mineral fertilizer (EO / EOM) comprising fermented plant growth promoting bacteria (BFCP) formulated in the solid state.   19. Fused EO / EOM fertilizer according to claim 17 or 18, wherein the EO / EOM is as defined in claims 15 or 16.   20. Fused EO / EOM fertilizer according to claim 17, 18 or 19 such that the BFCP is as defined in any one of claims 2 to 5.   21. Process for the fertilization of cultures comprising the application of EO / EOM bactericide according to any one of claims 17 to 20.   22. The method of claim 21 wherein the cultures are non-Leguminosea.   23. The method of claim 21 or 22 such that the application of the bactericidal fertilizer is carried out as a starter fertilizer.   24. The method of claim 21, 22 or 23 such that the application of the bacterized fertilizer is carried out in strips near the seeds.   25. A process according to any one of claims 21 to 24 such that the soils are free of large amounts of crop residues at the time of sowing.   26. A method of screening plant growth-promoting bacteria (BFCP) according to their hardiness, said BFCP being diazotrophic and MPS, comprising: - culturing precultures of BFCP at room temperature in a liquid medium free of nitrogen and phosphorus; phosphorus with the exception of hydroxypathy, - the application of osmotic pressure, - the incubation of cultures for a sufficient period of time, - the measurement of the optical density (OD) of the medium at two instants (DO1 and D02 ); - the hardiness of the bacteria depending on the difference (D02-D01).   27. A method for determining the bactericidal degree of biosolids useful for the manufacture of EO / EOMI bacterialized by plant growth-promoting bacteria (BFCP) comprising: - culturing precultures of BFCP in a liquid medium in the presence of said biosolids, incubation at room temperature, measurement of the optical density (OD) of the medium at two instants (OD1 5 and DO2), the bactericide of said biosolids being high when the difference (DO2DO1) is low or vice versa.