• 专利附录
  • 专利说明
  • 权利要求
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    • 专利摘要:
    The present invention relates to the treatment of seeds with bacteria of the Bacillus genus in order to improve their germination capacity and the vigor of the resulting seedling. The invention also describes a method to stimulate seed germination and the vigor of the resulting seedlings in soils or substrates with high salinity. Additionally, the invention also relates to the seeds obtained according to the methods for incorporating bacteria described above. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2792323A1
    申请号:ES201900081
    申请日:2019-05-06
    公开日:2020-11-10
    发明作者:Bejar Francisco De Borja Manuel Torres；Navarro Laura Toral；David Jonathan Castro
    申请人:Xtrem Biotech S L；
    IPC主号:
    专利说明:

    [0001] Microbiological method to improve seed germination
    [0002] Technical sector
    [0003] The present invention relates to a method of improving the germination rate of seeds and the vigor of the resulting seedlings by applying beneficial microorganisms of the genus Bacillus on the surface of the seed. Said method comprises applying a preparation comprising microorganisms to said seed. Furthermore, the present invention provides coated seeds that contain beneficial microorganisms.
    [0004] Background of the invention
    [0005] The demand for biological seed treatment solutions is increasing as they ensure that farmers protect their potential crop and quality while minimizing crop losses.
    [0006] It is well known that certain microorganisms when present in the soil, in the proximity of plant seeds, can work in symbiosis with seeds in a number of ways to enhance, for example, plant growth, or control of certain pests.
    [0007] These microorganisms known as rhizobacteria that live in soils in the vicinity of the roots. In many cases, these rhizobacteria benefit the ecosystem by stimulating plant growth and making agricultural production more sustainable. These microorganisms, also called growth-promoting rhizobacteria or PGPR (plant growth promoting rhizobacteria), colonize plant roots, compete with and control plant pathogens, and act as fertilizers.
    [0008] These bacteria have the ability to actively colonize the root system to promote and / or improve its growth and performance (Berendsen, Pieterse and Bakker, Trends Plant Sci. 2012 Aug; 17 (8): 478-86). The p G p R represent around 2 to 5% of rhizospheric bacteria (Jha and Saraf, Journal of Agricultural Research and Development 2015 Vol. 5 (2). Pp.
    [0009] 0108-0119,). The following genera of bacteria have been reported as PGPR: Agrobacterium, Arthrobacter, Azoarcus, Azospirillum, Azotobacter, Bacillus, Burkholderia, Caulobacter, Chromobacterium, Enterobacter, Erwinia, Flavobacterium, Klebsiella, Micrococcous, Pantoea, and Klebsemadiaretia (Rhizobhemadium and Serratiaretium , Journal of King Saud University - Science 2014, 26, 1-20).
    [0010] Bacterial inoculation to improve the productivity of different cultures has been practiced since the discovery of the beneficial effects of these bacteria. Methods used for the addition of these beneficial bacteria include seed coating, foliar application, and direct application to the soil.
    [0011] Most plants, and specifically those used by man as cultivated plants, use seeds to reproduce. However, on many occasions, the seeds after their maturation and dispersal are not able to germinate, either because they are dormant or because environmental conditions are not favorable for them. In this situation, the seeds begin to deteriorate, which is manifested by the progressive loss of their ability to germinate (that is, to improve their viability) and to give rise to healthy and vigorous seedlings. The vigor of a seed lot is defined as the set of properties that determine the level of activity and capacity of the seeds during germination and subsequent emergence of the seedlings.
    [0013] Given the importance of all these aspects in the field of seed physiology and technology, different protocols have been developed to improve the viability and vigor of the seeds, as well as to achieve storage conditions that ensure greater longevity.
    [0015] Biological seed treatment consists of active ingredients that can include microbes, such as fungi and bacteria, as well as plant and algae extracts. The seeds are treated with biological substances, in liquid or powder form. An even layer covers the entire seed.
    [0017] The biological treatment of seeds benefits the growth, the resilience of the plant, the abiotic stress, the development of the root system and the productivity of the crop. Among the advantages provided by the treatment of seeds with microorganisms, it is worth highlighting that it acts as a biostimulator and improves harvests, while helping plants to fight against pathogens and minimize biotic stress. Microorganisms colonize the roots and protect the crop throughout the growing season; improves the availability of nutrients in the root system and increases their assimilation. This better growth of roots and shoots means that early growth is optimized and both the nutritive value of the crop and the production yield are boosted.
    [0019] Biological seed treatment reduces the use of agrochemicals, which results in less exposure of farmers to chemicals and reduces their impact on the environment. In this way, the potential exposure of the soil to agrochemicals also decreases.
    [0021] Microorganisms used in seed treatment can be applied to the seed environment in a variety of ways. Generally, the bacteria can be applied directly to the soil, the seeds can be inoculated with the microorganisms immediately before sowing, or the seeds can be well coated with the microorganisms before sowing. One of the main difficulties encountered when using the latter approach is that most microorganisms are killed during the coating process.
    [0023] For coating seeds with microorganisms, most processes comprise the steps of making a suspension containing dry or non-dry microorganisms, applying the suspension to the seeds, and drying the coated seeds. Such a method, which generates seeds with microorganisms in the coating itself, is described for example in EP0818135. Another example of this approach is described in EP0097459, where the microorganisms are added so that they are approximately in the middle of a coating comprising a binder and filler. However, it was found that the presence of microorganisms in the aqueous suspension negatively affects their survival. Also, long drying times are needed for this type of application, eg 1.5 hours as described in EP0097459. This drying process consumes a lot of energy and has a negative impact on the viability of the seed, as well as on the microorganisms.
    [0025] Another option is to apply, for example, dry microorganisms to dry the coated or uncoated seed. For example, US6156699 describes alfalfa seeds that are sprayed with a mixture containing binder, limestone powder, surfactant, colorant, an agent suspension and 30.3% water by weight. After the addition of finely ground silica, the seeds are dried by a stream of hot air before applying a powder containing Rhizobium bacteria . A study by Brockwell and others. (Crop and Pasture Science, 1962, 13 (4): 638-649) compared such an approach with the suspension method described above. It was found that the incorporation of the microorganisms in the coating led to greater viability and stability of the inoculant than external application.
    [0027] Still another option is to coat the seeds with a binder and apply compositions containing dried organisms directly to them. For example, EP0192342 describes a dry bran carrier that is mixed with dry microorganisms that is applied to gum-coated seeds. EP0192342 is silent on the presence of minerals, it only leaves mineral particles. EP0494802 refers to the application of particles containing polymerized polysaccharides and dried microorganisms to seeds coated with an adhesive. US5041290 applies ascospore powder to adhesive coated seeds.
    [0028] EP3178325 describes the use of microorganisms of the species Bacillus methylotrophicus as plant growth stimulants and / or for the biological control of phytopathogens such as bacteria, insects, fungi and / or nematodes. In particular, EP3178325 describes the use of microorganisms belonging to Bacillus methylotrophicus strain XT1 (deposit number CECT8661) deposited on April 23, 2014 in the Spanish Collection of Type Cultures (CECT) by the University of Granada and / or microorganisms with a high degree of homology to strain XT1 whose DNA sequence is at least 99.6%, 99.7%, 99.8%, or 99.9% identical to the DNA sequence of strain XT1, based on the identity of all the nucleotides of said DNA sequences as plant growth stimulants and / or for the biological control of phytopathogens such as bacteria, insects, fungi and / or nematodes.
    [0029] EP3178325 also describes a method for stimulating plant growth comprising the steps of obtaining the bacteria, bacterial cultures or compositions and putting a plant in contact with the obtained bacteria, bacterial cultures or compositions. The procedure described in EP3178325 refers to already established plants that are in the growth phase.
    [0031] Despite numerous attempts to develop strategies for the treatment of seeds, there is currently an urgent need to develop alternative strategies or better strategies than the current ones, capable of promoting by biological procedures the germination of vegetable seeds and the vigor of the seedlings during the first days of development.
    [0033] The authors of the present application have surprisingly found that the application of Bacillus methylotrophicus strain XT1 (deposit number CECT8661) on seeds of various plant species improves the germination capacity of said seeds and the vigor of the seedling during the first days of developing. This bacterium has surprising characteristics that make it especially suitable for treating seeds. The bacterium is capable of producing exopolysaccharides with which it adheres to the seed without the need to incorporate adjuvants. On the other hand, the bacterium produces various metabolites that favor the germination and growth of the seedling during the first days of development. Additionally, the bacterium has antifungal capacity, preventing the infection of the seeds by phytopathogenic fungi.
    [0034] Explanation of the invention
    [0036] The present invention relates to the treatment of seeds with bacteria of the Bacillus genus in order to improve their germination capacity and the vigor of the resulting seedling. More specifically, the present invention relates to the use of microorganisms of the species Bacillus methylotrophicus for the treatment of vegetable seeds.
    [0038] Preferably, the present invention relates to the use of microorganisms of the species Bacillus methylotrophicus to improve the germination capacity and the vigor of the seedling resulting from seeds of the plant genera Asparagus, Brassica, Glycine, Solanum, Cucumis, Zea and Arabidopsis. More specifically, the present invention relates to the use of microorganisms of the species Bacillus methylotrophicus for the treatment of seeds of the species Asparagus officinalis, Brassica napus, Glycine max, Solanum lycopersicum, Cucumis sativus, Zea mais or Arabidopsis thaliana.
    [0040] Preferably, the present invention refers to the use of microorganisms of the species Bacillus methylotrophicus strain XT1 (deposit number CECT8661) deposited on April 23, 2014 in the Spanish Collection of Type Cultures (CECT) by the University of Granada to improve the capacity germination and the vigor of the seedling resulting from vegetable seeds.
    [0041] More specifically, the present invention refers to the use of microorganisms of the species Bacillus methylotrophicus strain XT1 (deposit number CECT8661) to improve the germination capacity and vigor of the seedling resulting from seeds of the plant genera Asparagus, Brassica, Glycine, Solanum , Cucumis, Zea and Arabidopsis. More specifically, the present invention relates to the use of microorganisms of the species Bacillus methylotrophicus strain XT1 (deposit number CECT8661) for the treatment of seeds of the species Asparagus officinalis, Brassica napus, Glycine max, Solanum lycopersicum, Cucumis sativus, Zea mais or Arabidopsis thaliana.
    [0043] The present invention also has as its object a method to stimulate the germination of seeds and the vigor of the resulting seedlings that comprises the steps of:
    [0044] to. Obtain a bacteria preparation as described above; Y
    [0045] b. Contacting a seed preparation with the bacterial preparation obtained in step a.
    [0046] c. Sow the seeds thus treated in a suitable substrate.
    [0048] Preferably the present invention describes a method for stimulating seed germination and the vigor of the resulting seedlings comprising the steps of:
    [0049] to. Obtain a bacteria preparation as described above; Y
    [0050] b. Putting a seed preparation in contact with the bacteria preparation obtained in step a in which the ratio of bacteria to the quantity of seeds is between 1 x 106 CFU (Colony Forming Units) of batteries per gram of seeds and 1 x 109 CFU of batteries per gram of seeds.
    [0051] c. Sow the seeds thus treated in a suitable substrate.
    [0053] The present invention also has as its object a method to stimulate the germination of seeds and the vigor of the resulting seedlings in soils or substrates with high salinity that comprises the steps of:
    [0054] to. Obtain a bacteria preparation as described above; Y
    [0055] b. Contacting a seed preparation with the bacterial preparation obtained in step a.
    [0056] c. Sow the seeds thus treated in a suitable substrate or soil with high salinity.
    [0057] Another object of the present invention is the seeds obtained according to the processes for incorporating bacteria described above. More specifically, the present invention includes the seeds treated with bacteria of the Bacillus genus , in particular it includes the seeds treated with bacteria of the Bacillus methylotrophicus species and more specifically, the present invention has for its object the seeds treated with microorganisms of the Bacillus methylotrophicus XT1 strain ( deposit number CECT8661).
    [0058] Brief description of the drawings
    [0059] Figure 1. Electron microscope image showing bacteria adhering to rapeseed. Upper left panel: untreated seeds (image at 3,000x magnification); Upper right panel: seeds treated with the XT1 bacteria (image at 3,000 magnification); Lower left panel: untreated seeds (image at 10,000X magnification); Lower right panel: seeds treated with the XT1 bacteria (image at 10,000 magnification). In the treated seeds, the presence of the bacterium XT 1 adhered to the surface of the seed is observed.
    [0060] Figure 2. Percentage of germination of Arabidopsis thaliana seeds in substrates with increasing concentrations of sodium chloride. White bars: seeds treated with water without bacteria (control); black bars: seeds treated with XT1 bacteria;
    [0061] Figure 3. Metabolic activity of Arabidopsis thaliana seeds in substrates with increasing concentrations of sodium chloride. Dashed line: seeds treated with water without bacteria (control); solid line: seeds treated with the XT1 bacteria.
    [0062] Preferred embodiment of the invention
    [0063] The present invention relates to the treatment of seeds with bacteria of the genus Bacillus. Example 1. Preparation of the bacteria suspension and treatment of the seeds.
    [0064] From a colony of the bacterium previously cultivated in Petri dishes, a culture was prepared in liquid medium (nutritive broth) shaking at 120 rpm for 48 hours at 28 ° C. Once the culture had grown, it was centrifuged at 14,000 g for 2 minutes and the supernatant was discarded. The bacterial pellet was resuspended in volume V 2 of the initial volume of sterile distilled water. This suspension was centrifuged again at 14,000 g for 2 minutes and the supernatant was discarded. This raising procedure to remove the remains of the culture supernatant was repeated 2 more times. After the last wash, the bacterial pellet was resuspended in sterile distilled water keeping a ratio of 1/10 of the initial volume.
    [0065] The number of microorganisms present in the suspension (CFU / ml, Colony Forming Units per milliliter) was then determined using serial dilutions and sowing on nutrient agar.
    [0066] Once the initial concentration of microorganisms in the suspensions was determined, it was adjusted with sterile distilled water to the desired concentration (between 108 and 109 CFU / ml depending on the case).
    [0067] These bacterial suspensions were mixed with the seeds using different proportions (between 104 and 109 CFU / g of seed depending on the case). For this the seeds were mixed with the bacterial suspensions and were kept at 28-30 ° C under stirring at 120 rpm for at least 1 hour.
    [0068] In order to determine the percentage of retention of the suspension of bacteria in the seeds, three seeds were taken for each of the treatments and a count of the microorganisms present on the surface of the seeds was made. For this, the seeds are washed with sterile saline solution and the number of microorganisms present in the washing solution is determined using serial dilutions and sowing on nutrient agar.
    [0069] Example 2. Treatment of asparagus seeds ( Asparagus officinalis).
    [0070] The asparagus seeds underwent a pre-treatment to activate synchronization in emergence. They were first stored at 4 ° C for seven days. Subsequently, the seeds were kept in water at 35 ° C for 12 hours and then they were disinfected by means of a 10-minute wash in 30% bleach and four washes in distilled water, each for 10 minutes.
    [0071] 53 activated and sterilized seeds were taken and treated as described in example 1 and subsequently sown in vegetable peat. As negative control, 53 activated and sterilized seeds were used that were treated with sterile water.
    [0072] At 15 days and 30 days, the percentage of live seedlings over the total number of planted seeds (% germination) and the vigor index (proportion of live seedlings over the total number of planted seeds multiplied by the height in centimeters of the aerial part of the seedling).
    [0073] The results obtained are summarized in Table 1:
    [0075]
    [0077] The results showed a clear increase in the percentage of germination and vigor of the resulting seedlings in the seeds treated with XT1 compared to those not treated both at 15 and 30 days post-sowing.
    [0078] Example 3. Treatment of rapeseed ( Brassica napus) seeds .
    [0079] The treatment of three samples of 2.5 g of sterilized rapeseed was tested with bacteria and inoculated with water (negative control), with a ratio of 1.25 x 106 CFU / g of seeds and with 1.25 x 107 CFU / g of seeds. The mixture was kept stirring for 30 minutes and 12 seeds from each group were later sown in 12-cell seedbeds with sterile sand / vermiculite.
    [0080] At 10, 15 and 25 days, the percentage of live seedlings over the total number of planted seeds (% germination) and the vigor index (proportion of live seedlings over the total number of planted seeds multiplied by the height in centimeters of the aerial part of the seedling). The results obtained are summarized in Table 2:
    [0082]
    [0084] The results showed a clear increase in the germination and vigor percentage of the resulting seedlings in the seeds treated with XT1 compared to the untreated ones at 10, 15 and 25 days post-sowing.
    [0085] Example 4. Treatment of soybeans ( Glycine max).
    [0086] The treatment of three samples of 10 g of soybeans was tested with bacteria and inoculated with water (negative control), with a proportion of 1.25 x 106 CFU / g of seeds and with 1.25 x 107 CFU / g of seeds. The mixture was kept stirring for 30 minutes and then 50 seeds from each group were sown in seedbeds with sterile sand / vermiculite.
    [0087] After 4 days in the dark, the seeds were kept for an additional 3 days with light / dark cycles (14/10). After this period, the percentage of live seedlings over the total planted seeds (% germination) and the vigor index (proportion of live seedlings over the total number of planted seeds multiplied by the height in centimeters of the aerial part of the seedling) were analyzed. ).
    [0088] The results obtained are summarized in Table 3:
    [0090]
    [0092] The results showed a clear increase in the percentage of germination and vigor of the resulting seedlings in the seeds treated with XT1 compared to the untreated ones.
    [0093] Example 5. Treatment of tomato seeds ( Solanum lycopersicum).
    [0094] The treatment of 14 tomato seeds was tested with bacteria and inoculated with water (negative control), with a ratio of 1.25 x 106 CFU / g of seeds and with 1.25 x 107 CFU / g of seeds. The mixture was kept stirring for 30 minutes and then 50 seeds from each group were sown in seedbeds with sterile sand / vermiculite.
    [0095] After 4 days in the dark, the seeds were kept for an additional 3 days with light / dark cycles (14/10). After this period, the percentage of live seedlings over the total planted seeds (% germination) and the vigor index (proportion of live seedlings over the total number of planted seeds multiplied by the height in centimeters of the aerial part of the seedling) were analyzed. ).
    [0096] The results obtained are summarized in Table 4:
    [0098]
    [0100] The results showed a clear increase in the percentage of germination and vigor of the resulting seedlings in the seeds treated with XT1 compared to the untreated ones.
    [0101] Example 6. Treatment of cucumber seeds ( Cucumis sativus).
    [0102] The treatment of 14 cucumber seeds was tested with bacteria and inoculated with water (negative control), with a ratio of 1.25 x 106 CFU / g of seeds and with 1.25 x 107 CFU / g of seeds. The mixture was kept stirring for 30 minutes and then 50 seeds from each group were sown in seedbeds with sterile sand / vermiculite.
    [0103] After 4 days in the dark, the seeds were kept for an additional 3 days with light / dark cycles (14/10). After this period, the percentage of live seedlings over the total planted seeds (% germination) and the vigor index (proportion of live seedlings over the total number of planted seeds multiplied by the height in centimeters of the aerial part of the seedling) were analyzed. ).
    [0104] Likewise, the mean wet weight of the seedlings was determined 17 days after germination.
    [0105] The results obtained are summarized in Table 5:
    [0107]
    [0109] The results showed a clear increase in the percentage of germination and vigor of the resulting seedlings in the seeds treated with XT1 compared to the untreated ones.
    [0110] Example 7. Treatment of corn (Zea mais) seeds .
    [0111] The treatment of 10 corn seeds that were inoculated with water (negative control) or with a ratio of 1.25 x 107 CFU / g of seeds was tested with bacteria. The mixture was kept stirring for 30 minutes and subsequently the 10 seeds of each group were sown in seedbeds with sterile sand / vermiculite.
    [0112] After 30 days, the percentage of live seedlings over the total number of planted seeds (% germination) and the dry weight of the aerial part and the roots were analyzed.
    [0113] The results obtained are summarized in Table 6:
    [0115]
    [0117] The results showed a clear increase in the germination and vigor percentage of the resulting seedlings in the seeds treated with XT 1 compared to the untreated ones.
    [0118] Example 8. Treatment of Arabidopsis thaliana seeds .
    [0119] Arabidopsis thaliana seeds were sterilized, according to the protocol detailed in previous reports. The sterilized seeds were divided into two groups: control and treated with the XT1 bacteria. For this last group, they were treated with 1.25 x 106 CFU / g of seeds and kept stirring at 28 ° C for 1 hour. After this time, the seeds of both treatments were resuspended in Hepes 0.1% agarose buffer.
    [0120] From each of the treatments, 50 µl of the seed suspension were taken to inoculate each of the wells (between 8 and 12 seeds per well), later the volume was made up to 100 µl with sterile water. The plate was incubated for 48 h in the dark at room temperature (20 ° C), and incubated for another 24 h in the dark. The percentage of germinated seeds was determined by microscopic observation.
    [0121] The results obtained are summarized in Table 7:
    [0123]
    [0125] The results showed a clear increase in the germination percentage in the seeds treated with XT1 compared to the untreated ones.
    [0126] Example 9. Seed treatment under high salinity conditions.
    [0127] Ambidopsis thaliana seeds were sterilized, according to the protocol detailed in the previous examples. The sterilized seeds were divided into two groups: control and treated with XT1 (1.25 x 106 CFU / g of seeds) that were kept stirred at 28 ° C for 1 hour. After the established time, the seeds of the two treatments were resuspended in Hepes 0.1% agarose buffer. From each of the treatments, 50 µl of the seed suspension were taken to inoculate each of the wells with 96-well microtiter plates, leaving between 6 and 12 seeds per well.
    [0128] The seeds of both groups were subjected to increasing concentrations of NaCl (0, 10, 20, 30, 40, 50, 60, 70, 80 and 90 mM). For this, the volume of each well was completed up to 100 µl with sterile water or with the corresponding NaCl solution to achieve the desired salt concentration. Two repetitions were performed for each of the indicated salt concentrations.
    [0129] The plate was incubated for 72 h in the dark at 21 ° C and the germination percentage was determined by observation with the binocular magnifying glass.
    [0130] The results obtained are represented in Table 8 and in Figure 2:
    [0132]
    [0133] The results showed a clear increase in the germination percentage in the seeds treated with XT1 compared to the untreated ones in the entire NaCl concentration range in the germination medium.
    [0135] Indirect methods for quantifying the metabolic activity of seeds based on the spectrophotometric measurement of the reduction of methyl-tetrazolium salts such as the 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT). The metabolic activity of seed cells includes that of dehydrogenases. The activity of mitochondrial dehydrogenases, in particular succinate dehydrogenase, but cytosolic reductases or other subcellular compartments can also intervene, act on MTT and the resulting reduced coenzymes (NADH and NADPH) convert MTT to its formazan derivative. This violet colored derivative is insoluble in water. To quantify it, it is dissolved in an organic solvent, such as DMSO (dimethylsulfoxide) and absorbance at 590 nm is measured.
    [0137] The seeds of both groups (treated with XT1 and control treated with water) were subjected to increasing concentrations of NaCl (0, 10, 20, 30, 40, 50, 60, 70, 80 and 90 mM). For this, the volume of each well was completed up to 100 µl with sterile water or with the corresponding NaCl solution to achieve the desired salt concentration. Two repetitions were performed for each of the indicated salt concentrations.
    [0139] The plate was incubated for 72 h in the dark at 21 ° C and after that period 10 µl of an MTT solution were added and incubated for another 24 h in the dark. Finally, 100 µl of lysis buffer were added to solubilize the salt deposits produced by the MTT. After another 24 h in the dark, the absorbance at 590 nm of each well was determined.
    [0141] The results represented in figure 3 show a clear increase in the metabolic activity of the seeds treated with XT1 compared to the untreated ones in the whole range of NaCl concentration in the germination medium.
    权利要求:
    Claims (16)
    [1]
    1. A procedure to increase the germination rate of seeds and the vigor of the resulting seedlings by applying beneficial microorganisms to the surface of the seed.
    [2]
    2. A method according to claim 1 wherein said microorganisms belong to the genus Bacillus.
    [3]
    3. A method according to claim 1 wherein said microorganisms belong to the species Bacillus methylotrophicus.
    [4]
    4. A method according to claim 1 in which said microorganisms belong to the Bacillus methylotrophicus strain XT1 (CECT8661).
    [5]
    5. A process according to claim 4 in which said the proportion of microorganisms added is in the range between 104 and 109 CFU / g of seed.
    [6]
    6. A process according to claim 5 in which said the proportion of microorganisms added is in the range of 106 and 108 CFU / g of seed.
    [7]
    A method according to claims 1 to 6 in which the seeds are from monocotyledonous plants.
    [8]
    8. A process according to claims 7, characterized in that the seeds belong to the genera Asparagus, Saccharum, Avena, Triticum, Zea, Oryza, Secale, Hordeum.
    [9]
    9. A method according to claims 1 to 6 wherein the seeds are from dicotyledonous plants.
    A process according to claim 9, characterized in that the seeds belong to the genera Brassica, Solanum, Capsicum, Cucumis, Phaseolus, Glycine, Arabidopsis, Chenopodium.
    [10]
    10. Vegetable seeds characterized by containing on their surface microorganisms of the Bacillus genus obtained according to the procedure described in claim 2.
    [11]
    11. Vegetable seeds characterized by containing on their surface microorganisms of the Bacillus methylotrophycus species obtained according to the procedure described in claim 3.
    [12]
    12. Vegetable seeds characterized by containing on their surface microorganisms of the strain Bacillus methylotrophycus XT1 (CECT8661) obtained according to the procedure described in claim 4.
    [13]
    13. A procedure to increase the germination rate of seeds in conditions of high salinity of the substrate by applying beneficial microorganisms on the surface of the seed.
    [14]
    14. A method according to claim 13 wherein said microorganisms belong to the genus Bacillus.
    [15]
    15. A method according to claim 13 wherein said microorganisms belong to the species Bacillus methylotrophicus.
    [16]
    16. A method according to claim 13 wherein said microorganisms belong to the Bacillus methylotrophicus strain XT1 (CECT8661).
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    同族专利:
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    ES2792323B2|2022-01-19|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    ES2561908A1|2014-07-31|2016-03-01|Universidad De Granada|Use of bacillus methylotrophicus as a plant growth stimulant and biological control medium, and isolated strains of said species |
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    申请号 | 申请日 | 专利标题
    ES201900081A|ES2792323B2|2019-05-06|2019-05-06|Microbiological method to improve seed germination|ES201900081A| ES2792323B2|2019-05-06|2019-05-06|Microbiological method to improve seed germination|
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