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    • 专利摘要:
    Product for the control of phytopathogenic fungi causing diseases of vine wood and procedure for its application in vine grafts. There are 3 strains of actinobacteria belonging to the genus streptomyces and called streptomyces sp.vv/r1, streptomyces sp. Vv/r4 and streptomyces sp. Vv/e1. Strains vv/r1 and vv/r2 have been isolated from vine rhizosphere, while strain vv/e1 is an endophytic strain isolated from the root interior, in both cases obtained from vine plants of an age of 1 year. These strains were selected on the basis of their ability to effectively inhibit the growth of the phytopathogenic fungi llyonectria sp., phaeomoniella chlamydospora and phaeoacremonium aleophilum in in vitro tests and vine grafts in the nursery, as indicated by numerous studies as the main causes of decay. Of young vine plants, which can be infected through the root system. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2543363A1
    申请号:ES201530448
    申请日:2015-04-01
    公开日:2015-08-18
    发明作者:José Manuel ÁLVAREZ PÉREZ；Sandra GONZÁLEZ GARCÍA；Rebeca COBOS ROMÁN；Miguel Ángel OLEGO MORÁN；José Enrique GARZÓN JIMENO；Juan José RUBIO COQUE
    申请人:VIVEROS VILLANUEVA VIDES S L；VIVEROS VILLANUEVA VIDES SL；
    IPC主号:
    专利说明:

    OBJECT OF THE INVENTION
    The present invention relates to the selection of vine rhizosphere actinobacteria (Vitís vinífera) or endophytic actinobacteria isolated from the inside of the root of
    vine plants to reduce the degree of fungal infection causing
    Vine wood diseases through the root system by applying to vine grafts before being established in the nursery.
    The object of the invention is to provide an economical technology, of easy
    application, and that can be applied during the industrial production process of
    vine plants in nursery, or even that can be applied to adult vineyards and
    established in the field in order to avoid or reduce the risk of infection through
    root system by phytopathogenic fungi that cause diseases of grapevine wood.
    5 The method of the invention finds special application in plant nurseries
    vine in order to produce plants with a better sanitary status or in dedicated companies
    to the production of grapes, both table and dedicated to the production of wine.
    According to the method, a decrease in the levels of
    10 infection of treated plants ranging from 73.1% to 76.9% by phytopathogenic fungi Ilyonectria sp., Phaeomoniella chlamydospora and Phaeoacremonium aleophilum, when compared with the level of infection detected in grapevine grafts
    untreated control with actinobacteria.There has also been a decrease in the mortality of vine grafts in the
    15 establishment phase of the nursery on land for the production of vine plants ranging between 5.4% and 12.4% by comparison with the degree of mortality observed in lots of untreated control plants.
    BACKGROUND OF THE INVENTION Since the early 90s of the twentieth century the symptoms of vine wood diseases in young vineyards (vine decay), as well as failures in the
    Planting material has been assuming significant economic losses worldwide for the wine sector. It is currently considered that fungi
    Causes of vine wood diseases such as black foot, disease
    Petri or syndromes produced by species of the Botryosphaeriaceae family are one of the factors that contribute most significantly to decay
    of young vine plants [Gramaje and Armengol, 2011, Plant Dis. 95, 1040-1055]. 10 Among the symptoms observed are low yields of berry production, rickets and poor development and poor development of the apparatus
    radicular with the appearance of necrotic lesions, both in root and in branches. These symptoms lead to a marked delay in plant growth and with high frequency in premature death.
    15 The main pathologies associated with the decay of young vine plants are the black foot, produced by different species of the genus Ilyonectria (formerly Cylindrocarpon) [Halllen et al., 2006, Phytopathol. Mediterr. 45, S55-S67] and Petn's disease, mainly caused by Phaeomoniella chlamydospora fungi and different species of the Phaeoacremonium genus [Mundy and Manning, 2010, New Zealand Plant Protection 63: 160-166].
    5 Different studies have shown that many of the affected plants had been infected in the nursery during the different stages of industrial production of vine plants. In fact, several moments or stages of the process have been identified in which the contamination of the material used for the production of vine plants can occur, mainly 1) .- the infection of the plants
    10 mother of American varieties, from which the rootstocks are produced; 2) .- infections produced during the grafting and callus formation stages between the rootstock and the variety; 3) .- infections produced during the establishment phase of the nursery on land [Gramaje and Armengol, 2011, Plant Dis. 95, 1040-1055].
    Infection during the propagation process in the nursery is a very important problem, since in this way pathogens are introduced into the cycle of
    Plant production The role of vine nurseries as the origin of the contamination of a large number of plants has been demonstrated by Arcca
    et al. [Aroca et al., 2006, Eur. J. Plant Pathol. 115: 195-202] that in a screening carried out on grafted plants and patterns of American varieties they found that 45.2% of the plants were infected with some fungus associated with 5 decay of the vine, especially the species responsible for the disease
    of Petri.
    In another study [Giménez-Jaime et al., 2006, J. Phytopathol. 154: 598-602] carried out in 6 nurseries of the Valencian Community verified the presence of 10 fungi causing wood diseases in all nurseries analyzed. From
    more concrete form Cylindrocarpon spp. Botryospheria spp. and B. obtusa (O. seriata) they were preferably isolated in plant material corresponding to early phases
    of the production process (patterns of the American variety and graft area in grafted plants before being planted in soil) in percentages ranging from
    15 0.48-2.38% of the plants analyzed. On the contrary P. chlamydospora and P. aleophilum were not detected until the cuttings were planted in the field to develop roots and shoots (aerial part), with infection percentages ranging between 0-7.62% of the analyzed plants.
    The problem is not exclusive of nurseries in Spain. In fact Cy / indrocarpon spp. it has been isolated in South Africa from the mother plants of the graft carriers and in this country and California this genus is considered the most important pathogen in nurseries
    5 [Hallen et al., 2003, Australas. Plant Pathol 32: 47-52 .; Fourie and Hallen, 2004, Australasian Plant Pathol. 33: 313-315; Dubrovsky and Fabritius, 2007, Phytopathol. Mediterr. 46: 46-86].
    The most important phytopathogenic fungi indicated as responsible for
    10 decay of young vine plants have in common their ability to infect plants through the root system. This has demonstrated the presence of fungi responsible for black foot disease in the soil [Agustí-Brisach et al. , 2014, Plant Pathol. , 63: 316-322]. Fungi can remain here for long periods of time (in the form of clamidospores or in the form of
    15 saprophytic) and penetrate the plant through the roots, already from the phase of
    rooting in nursery. Proper soil management can significantly reduce the risk of infection through the root. It has also
    demonstrated the ability of P. ch / amydospora and P a / eophilum to use this route of entry to infect grafts. In fact, Giménez-Jaime et al. [Giménez-Jaime et al., 2006, J. Phytopathol. 154: 598-602] have described that these two species could only be isolated from grafts two months after being
    planted in the nursery. All these studies confirm that these pathogenic fungi 5 can infect the grafts through the root apparatus that develops in the nursery.
    In recent years numerous studies have been developed to reduce
    the risk of infection of vine plants in nursery through the use of different
    10 strategies such as the use of different chemical fungicides, treatments of plant material with hot water, biocontrol or the development of varieties of rootstocks more resistant to infection [for a review see Gramaje and Armengol, 2011, Plant Dis. 95, 1040-1055].
    15 Biocontrol is one of the most promising technologies for the control of many pathologies without having to resort to the use of pesticides of chemical synthesis, of high toxicity in many cases and that have frequently been shown to be harmful to the environment and / or living beings. In vine nurseries they have
    performed different tests with biocontrol agents (SeA). Thus Fourie and collaborators showed that the application of different strains of the Trichoderma fungus had a positive effect to prevent infection by Cy / indrocarpon spp., P. ch / amydospora and Phaeoacremonium spp. In fact levels were observed
    5 minor of these fungi in the roots of plants treated in nursery [Fourie et aL,
    2001, Phy1opathol. Mediterr. 40S: 473-478]. It has also been tested in SeA Trichoderma halZianum vine nurseries to control P. chlamydospora infection. He
    SCA was applied in different ways: by immersion of the root mass in
    a spore suspension, by immersion of the stems that are grafted on the 10 foot of the plant to form the callus in spore suspension or by a
    combination of both. The ACS showed some efficacy by promoting the development of the root system and increasing the percentage of plants that survived infection with the pathogen. However, also as an adverse effect was observed
    an increase in the mortality of plants treated with SeA [De Marco and Osti,
    15 2007, Phy1opathol. Mediterr. 46: 73-83], so the effectiveness of the application of seA of the genus Trichoderma remains in doubt.
    DESCRIPTION OF THE INVENTION
    The technology that is recommended is designed to solve a specific point of the aforementioned problem, specifically reducing or limiting to the maximum the infection of vine grafts in the nursery through the root apparatus during the phase
    of the establishment of the ground graft nursery, at which time these
    develop the root system For this we rely on employment as agents
    Biological control (BeAs) of 3 selected strains of actinobacteria, when applied to grapevine grafts
    A) .- Streptomyces sp. VV / E1, W / R1 and W / R4: description.
    In one aspect the invention relates to 3 strains of actinobacteria.
    belonging to the genus Streptomyces sp. selected from the rhizosphere (strains VV / R1 and VV / R4) or from the inside of the roots (strain VV / E1) of vine plants aged 15 years of 1 year and showing high effectiveness to inhibit growth, both
    in in vlfro tests as in plant material, of phytopathogenic fungi
    Iyonectria sp., P. chlamydospora and P. aleophilum, mainly responsible for
    decay of young grapevine plants and various diseases of grapevine wood.
    The respective cultures of the Streptomyces sp. denominated VV / E1, W / R 1 and VV / R4 they are conserved and deposited of permanent form in the Spanish Collection of Cultures Type (CECT), located in University of Valencia;
    5 Research building; Burjassot Campus; 46100-Burjassot (Valencia), with the respective access numbers CECT 8852; CECT 8853 and CECT 8854.
    Streptomyces sp. W / E1, VV / R1 and VV / R4 present among others the
    Following features listed below:
    1) .- Strain W / E1.-Sequencing of ribosomal DNA (rDNA) 16S concluded that it is a Streptomyces strain belonging to the taxonomic group S capil / ispiralis-S cell / ulosae-S gancidicus-S pseudogriseolus-S werraensis, being indistinguishable based on the 16S rDNA sequence, since it shows a homology
    15 of 100%.
    The W / E1 strain has good growth in the ISP1, ISP2, ISP3, ISP4, ISP5, ISP6, MBA, MEY, TBO and 2xTY media. This strain has a very low capacity to
    sporulation in ISP3 and TBO media, not sporulating in ISP1, ISP2, ISP4, ISP5, ISP6, MBA, MEY, Y2XTY media.
    Morphologically in medium plates ISP2 presents colonies with light brown coloration, rough or striated morphology and with a bulge in the center.
    11) .- Strain VV / R1.-The sequencing of ribosomal DNA (rDNA) 16S concluded that it is a strain of Slreplomyces belonging to taxonomic group 5. peucelius-5o kursanovi-5o xanlholilicus, being indistinguishable based on the sequence
    10 of the 16S rDNA, since it shows a 100% homology.
    The VV / R1 strain has good growth in the ISP1, ISP2, ISP3, ISP5, ISP6, MBA, MEY, TBO and 2xTY media, showing lower growth in ISP4 medium. This strain has a low sporulation capacity in MEY medium, not sporulating
    15 apparently in ISP1, ISP2, ISP3, ISP4, ISP5, ISP6, MBA, TBO and 2xTY media.
    In ISP2 medium plates morphologically characterized by presenting a pinkish coloration, with rough morphology, some concentricity in its growth
    with a bulge in the center.
    11) .- Strain VV / R4.-Sequencing of the ribosomal AON (AONr) 16S concluded that it is a Streptomyces strain belonging to the taxonomic group S
    5 capil / ispiralis-S cel / ulosae-S gancidicus-S pseudogriseolus-S werraensis, being indistinguishable based on the AONr 16S sequence, as it shows 100% homology.
    The strain VV / R1 has good growth in the media ISP1, ISP2, ISP3, ISP4, ISPS,
    10 MBA, MEY, TBO AND 2xTY, showing lower growth in ISP6 medium. This strain has a very low sporulation capacity in ISP4 medium; low sporulation capacity between ISP1, ISP7 and MBA media; average sporulation capacity in ISP3 and MEY; high sporulation capacity between ISP2 and TBO. It does not show sporulation capacity in ISPS and 2xTY media.
    Morphologically in ISP2 medium plates it is characterized by producing colonies of
    a dark beige color, with flat morphology and concentric growth. Tend to
    sporular in this medium (white color).
    B) .- Isolation of Streptomyces sp. VV / R1, VV / R4 and VV / E1.
    The mentioned strains have been isolated and selected from a total of 85
    strains of rhizospheric actinobacteria and 66 endophytic strains analyzed. These
    5 strains were isolated from samples of root rhizosphere in half starch and casein or from inside of vine root, according to the methodology described by Li et al. [Li et al., 2012, Ant. Van Leeuwenhoek. 101: 515-527]. Samples of rhizosphere soil or root material were successively diluted in
    saline (0.9% NaCl) sterile. Dilutions were plated in medium with
    10 starch and casein containing cycloheximide and nalidixic acid to inhibit
    selectively the growth of fungi and Gram (-) bacteria respectively. The
    plates were incubated 7 days at 28 ° C. Then all the colonies with a typical
    Actinobacterial morphology (mycelial development, velvety or waxy appearance, possible production of aerial mycelium with or without exospores and possibility of production of dilusible pigments in the culture medium) were replicated to half yeast extract-malt extract (MEY) [ Hopwood et al., 1985, Genetic manipulation 01 Streptomyces-A laboratory manual. The John Innes Foundation, Norwich, UK and Cold Spring Harbor Laboratory] .. Provisional confirmation of your membership in the
    Actinobacteria group was performed by determining their filamentous morphology after observation under an optical microscope.
    C) .- Selection of Streptomyces sp. VV / R1, VV / R4 and VV / E1.Strains W / R1, VV / R4 and VV / E1 can be used to obtain plants
    5 vine grafted into vine nurseries with a medium level of fungal infection
    causing wood diseases (particularly / lyonectria sp., P. ch / amydospora and P. a / eophi / um) much lower. This application would allow the
    applicant company have a plant material with a very sanitary state
    improved.
    These strains can be used individually or jointly, in different
    combinations, for application in vine grafts by inclusion in the
    rooting hormone solution or for application to young and adult plants in the field by application to the root system by fertirrigation or
    15 other irrigation or viticultural techniques, in order to control the infection of vine plants by these phytopathogenic fungi through the root apparatus.
    The use of the W / R1, VV / R4 and W / E1 strains in obtaining vine plants
    Grafted provides numerous advantages. In particular its employment in nurseries of
    Vine allows the obtaining of plants with a better sanitary status by decreasing the level of fungal infection causing wood diseases. These plants, for
    5 their best health status, they will be less susceptible to suffer later
    infections once planted definitely in the field, especially if
    they make periodic contributions of these microorganisms to the root system through fertirrigation or by any other alternative technique. The implantation of these microorganisms in the rhizosphere of plants or inside the system
    10 root will prevent or limit infection by any fungus susceptible to the antifungal mechanisms or compounds produced by them.
    DESCRIPTION OF THE DRAWINGS
    15 To complement the description that will then be carried out and with a view to
    to help a better understanding of the features of the invention, according to a preferred example of practical realization thereof, is accompanied as
    integral part of this description, a set of drawings where with character
    Illustrative and not limiting, the following has been represented:
    Figure 1.- Shows a graph corresponding to the decrease in the average level of
    mortality observed in vine grafts established in nursery in the field after
    5 application of the selected actinobacteria VV / E1, VV / R 1 and VV / R4 by comparison with the average level of mortality observed in untreated control plants.
    10 Figure 2.- Shows a graph corresponding to the relative decrease ("lo) of pathogenic fungi lIyonectria sp., P. chlamydospora and P. aleophilum observed in vine grafts established in nursery in the field after the application of the selected actinobacteria W / E1, W / R1 and W / R4 by comparison with the level
    infection medium observed in untreated control plants.
    EXAMPLES OF PREFERRED REALIZATION OF THE INVENTION
    The following examples illustrate the invention, although they should not be considered as limiting the scope thereof.
    • Example 1. Obtaining and characterizing strains VV / R1, VV / R4 and VV / E1. A total of 85 strains of rhizospheric actinobacteria and 66 isolated endophytic strains
    of 1-year-old vine plants were differentiated according to easily appreciable morphological features 5 (color and morphology of the colony, ability to form
    aerial mycelium, ability to sporulate, soluble pigment production and
    diffusible in the culture medium). All these strains were tested for their ability to inhibit the growth of phytopathogenic fungi Ilyonectria sp. Y
    Serious diplodia due to its aggressive growth. For this, the technique of
    10 bioassay All strains were grown at 28 ° C in medium agar malt extract (MEA) plates for 5 days. Then an agar block containing mycelium of the phytopathogenic fungi mentioned above was placed at the center of the plate at a distance of 2 cm from the edge of the colony of the tested actinobacterium. The plates were then incubated at 25 ° C for 5-10 days to
    15 determine the possible inhibitory effect of the actinobacterial strain on phytopathogenic fungi. A total of 1 endophytic actinobacteria (W / E1) and 2 rhizospheric strains (W / R1 and W / R4) were selected for their ability to inhibit in vitro
    effectively the growth of both pathogens. Inhibition ability
    of growth was determined by the methodology of Lamsal et al. (2012) [Mycobiology 40: 244-251], calculating the Inhibition Rate (TI,%) of each of the actinobacteria using the following expression: TI (%) = [1- (Oa-0) / (Oc0) ] x1 OD; where Da is the growth of the fungus in the antagonism trial, Oc is 5 the diameter of growth in the control (no antagonism with any
    actinobacteria), and O is the diameter of the agar block containing the fungus (7 mm).
    This experiment was performed in triplicate for each of the actinobacteria. In this trial, the VV / E1 strain showed a growth inhibition against D. seriala of 56.8 (± 6.1)% and 38.8 (± 2.9)% against l. macrodldyma. VV / R1 strain
    10 showed respectively a growth inhibition of 48.8 (± 3.9)% and 53.7 (± 4.2)% against D. seriata and 1. macrodidyma respectively. The strain VV / R4 was able to inhibit in vi / ro the growth of D. seriala in a percentage of 51.7 (± 6.2) and that of l. macrodidymaen 50.8% (± 3.7).
    The 3 preselected strains also showed a high capacity to inhibit in
    15 vitro growth of other phytopathogenic fungi such as Cadophora luteo-olivacea, Eutypa lata, // yonectria sp., Phaeomonie // a chlamydospora and Phaeoacremonium aleophilum
    • Example 2. Biomass production of strains W / R1, W / R4 and W / E1. Two different methodologies were used to test the versatility of each
    strain when producing biomass.
    5 2.1) .- Biomass production due to growth in flasks in aerobic conditions.
    For this, 1 L flasks containing 0.2 L of half extract broth were used
    of malt Each flask was inoculated with 5 ml of a culture developed in it
    10 average of 0.0. 1 estimated spectrophotometrically at 600 nm. The flasks were incubated at 28'C with constant agitation of 220 rpm in an orbital incubator. He
    Growth was determined every 24 hours. During the process the homogeneity of the culture was verified by direct observation under the optical microscope. A
    Once the incubation was finished, cell viability was checked by counting 15 colony forming units per ml (CFU / ml). All strains grew with
    very quickly in these conditions obtaining a large amount of biomass and a viability equal to or greater than that of the species S. coe / ic% r used as
    control.
    2.2) .- Production of biomass in fermenter.
    5 The study was carried out in a 2.5 L Biostat-A laboratory fermenter, controlling the different parameters that affect strains such as 02 dissolved, pH, foam, acidity and temperature. For the development of the cultures, 1.5 L of medium malt extract broth was used which was inoculated with a preculture of each bacteria until a cell suspension of 0.0 is obtained. = 0.1 (determined at 600
    10 nm). The pH of the culture was kept constant at a value of 7.0 and the temperature of
    incubation was 28 ° C. Finally the concentration of dissolved oxygen is
    kept constant at 35%. Growth and cell viability were monitored
    as indicated in the previous section. Under these conditions all strains showed a fast and homogeneous biomass production with a high
    15 cell viability, of the same order of magnitude as the S. coelico! Or strain used
    As a Witness.
    • Example 3. Field trials: application of Streptomyces sp. W / R1, VV / R4 and VV / E1 to vine grafts.
    The 3 preselected actinobacteria strains - endophytic acinobacteria (W / E1)
    5 AND 2 rhizospheric strains (VV / R1 and W / R4) - malt extract was grown in half medium by inoculating 200 ml of medium in 1 L capacity flasks at 28'C and 220 rpm of agitation for 120 hours. Microbial biomass was precipitated by centrifugation at 12,000 xg for 20
    10 minutes at 4'C. It was then resuspended in 50 ml of 20% glycerol (w / v) and kept at -20 ° C until application.
    Immediately before application, a quantification of the number of bacteria present in each preparation was carried out by planting successive dilutions in
    15 POA medium and incubation at 28'C for 5 days.
    Bacteria were applied to the grafts on batches of 100 grafts (R-110 rootstock and white Tempranilla variety) by immersion of the basal portion of the rootstock in 2L of solution containing rooting hormone to which
    107 bacterial cells were added per ml in the form of a 20% glycerol suspension (w / v). In parallel, a negative graft control was performed to which an equivalent amount of 20% glycerol without bacteria was added.
    The grafts are kept in rooting solution with bacteria for 24 hours. After this time they are planted in the field for the establishment of an experimental nursery, which is maintained in optimal conditions by personnel
    10 specialized of Viveros Villa nueva Vides S.L. through the contribution of the program normal fertirrigation that the company applies.
    Seventy days after its establishment on the ground, a graft control was performed consisting of: determination of mortality level (non-sprouted grafts
    15 versus total grafts placed in the field), total height of the plant and length of
    Seventh internode.
    The plants were uprooted 200 days after their establishment in the field and they were
    proceeded to determine the level of infection by pathogenic fungi Ilyonectria sp, P. chlamydospora and P. aleophilum at the root insertion point, in order to determine the possible protective effect of the applied actinobacteria to avoid
    5 infection of these fungi through the root system. The analysis was performed of
    according to the methodology described by Giménez-Jaime et al. [Giménez-Jaime et al., 2006, J. Phytopathol. 154: 598-602 [And Aroca et al. [Aroca et al., 2006, Eur. J. Plant Pathol. 115: 195-202].
    10 The identification of the isolated fungi was carried out as follows:
    - To identify the species of the Botryosphaeriaceae family, the classical morphological criteria were followed [Phíllips et al., 2013, Studíes in Mycology 76: 51-167] and the molecular techniques described for this group [Alves et al., 2007, Res.
    15 Microbiol. 158: 112-21; Martos et al., 2009, Phytopathol. Mediterr. 48: 162; Spagnolo et al. 2011, Europ. J. Plant Palhol. 129: 485-500].
    - The different species of the genus Phaeoacremon; um were identified following the
    electronic identification key developed by Mostert et al. [Mostert et al. 2006, Studies in Mycology 54: 1-113) Additionally, specific oligonucleotides were used to detect P. a / eophi / um [Tegli et al., 2000,
    5 Phytopathol. Mediterr. 39: 134-49).
    - Identification of species of lithonetry was carried out taking into account the
    Recent reclassification of some species of the genus "Cylindrocarpon" [Cabral et al., 2012, Fungal biology 116: 62-80.) associated with wood diseases.
    - The identification of Phaeomonie // a ch / amydospora was developed according to the classic described morphological criteria [Crous and Gams, 2006, Phytopathol. Mediterr. 39: 112-18) And the specific detection by PCR [Tegli et al., 2000, Phytopathol. Mediterr. 39: 134-49).
    15 Field trials were conducted for 2 consecutive years in 3 lots of 25
    plants per treatment and year.
    Field trials showed that:
    A) .- Control grafts established in nursery field showed a medium level
    of mortality of 31.7%. The application of actinobacteria to the grafts produced
    5 a decrease in mortality to 26.3% -VV / E1-strain (decrease in mortality compared to 17.0% control); a decrease in mortality to 19.3% - W / R1 strain (decrease in mortality from the control of 39.1%) And a decrease in mortality to 21.0% - VV / R4 strain - (decrease in mortality from the 33.8% control) , as we can see in Figure 2.
    10 B) .- The degree of infection of the grafts established in the nursery in the field decreased significantly after the application of the selected actinobacteria (Figure 3). Indeed, assigning a relative value of 100 for the level of infection by pathogenic fungi Ilyonectria sp., P. chlamydospora and P. aleophilum in grafted control plants (not treated with actinobacteria) and established in nursery in
    15 field at the end of the growing season we observe how the level of infection
    decreased to 26.9% (decrease of 73.1%) in the case of plants treated with strains VV / R1 and VV / R4, while the level of infection decreased to 23.1% (decrease of 76.9%) in the case of plants treated with Actinobacteria W / E1.
    权利要求:
    Claims (1)
    [1]
    1 '.- Product for the control of phytopathogenic fungi causing diseases of
    vine wood and its application in vine grafts, characterized in that it materializes
    5 in the strain of the genus Streptomyces sp. and deposited in the CECT with the access number CECT 8852 (strain Streptomycessp. VV / E1).
    2 '.- Product for the control of phytopathogenic fungi that cause diseases of grapevine wood and its application in grapevine grafts, characterized in that it materializes in the strain of the genus Streptomyces sp. and deposited in the CECT with the access number CECT 8853 (strain Streptomycessp. VV / R1)
    3 '.- Product for the control of phytopathogenic fungi causing diseases of
    vine wood and its application in vine grafts, characterized in that it materializes
    15 in the strain of the genus Streptomyces sp. and deposited in the CECT with the access number CECT 8854 (strain Streptomycessp. VV / R4).
    4 '.- Procedure for the control of phytopathogenic fungi causing
    vine wood diseases, according to the products of claims 1 to 3, characterized in that it consists in the application individually, or in
    any combination of said products, to vine grafts during the phase of
    rooting hormone treatment (or any other phase prior to
    5 establishment of the graft nursery on land) to obtain vine plants with a lower degree of fungal infection causing diseases of
    vine wood
    5 '.- Procedure for the control of phytopathogenic fungi causing 10 diseases of vine wood, according to the products of the claims
    1 to 3, characterized in that it consists of applying individually or in any combination of said products to vine grafts, during the rooting hormone treatment phase (or any other phase prior to the establishment of the graft nursery on land) to get a reduction in the
    15 graft mortality level established in nursery.
    6 '.- Procedure for the control of phytopathogenic fungi causing
    vine wood diseases, according to the products of claims 1 to 3, characterized in that it consists in the application individually or in
    any combination of such products to the root apparatus of young plants and
    adults in the field through fertirrigation or any other alternative technique for
    5 control of fungi causing diseases of vine wood.
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    公开号 | 公开日
    ES2543363B2|2016-03-15|
    PT3075249T|2020-10-19|
    HUE051393T2|2021-03-01|
    EP3075249B1|2020-07-22|
    EP3075249A1|2016-10-05|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    WO2010115802A1|2009-03-31|2010-10-14|Universite De Reims Champagne-Ardenne|New actinomycetes strain compositions and their use for the prevention and/or the control of micro organism inducing plant diseases|CN111184031A|2018-11-15|2020-05-22|浙江省大盘山国家级自然保护区管理局|Liquid composite mycorrhizal inoculant for promoting growth of blueberry seedlings and preparation method thereof|ES2389343B2|2011-03-28|2013-06-06|Bodegas Vega Sicilia S.A.|USE OF NATURAL ANTIFUNGALS TO PREVENT INFECTION OF INJURIES, GRAFTING AND PRUNING WOUNDS IN WOODY PLANTS BY PHYTOPATHOGEN FUNGI.|
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    PT160007290T| PT3075249T|2015-04-01|2016-03-30|Product for controlling phytopathogenic fungi that cause grapevine wood diseases and method for the application thereof in grapevine grafts|
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