ES2387666A1 - Method for large-scale production of mycelial inoculum of ascomycota - Google Patents
Method for large-scale production of mycelial inoculum of ascomycota Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 15
- 241000235349 Ascomycota Species 0.000 title claims abstract description 11
- 239000002054 inoculum Substances 0.000 title abstract description 6
- 238000011031 large-scale manufacturing process Methods 0.000 title abstract description 5
- 230000012010 growth Effects 0.000 claims abstract description 21
- 239000001963 growth medium Substances 0.000 claims abstract description 19
- 239000002609 medium Substances 0.000 claims abstract description 18
- 241000233866 Fungi Species 0.000 claims abstract description 16
- 230000002538 fungal effect Effects 0.000 claims abstract description 15
- 244000223977 Tuber melanosporum Species 0.000 claims abstract description 11
- 241001279783 Terfezia Species 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 8
- 241001279770 Terfezia arenaria Species 0.000 claims abstract description 7
- 238000000746 purification Methods 0.000 claims abstract description 7
- 229960000304 folic acid Drugs 0.000 claims abstract description 4
- 239000011724 folic acid Substances 0.000 claims abstract description 4
- 238000011081 inoculation Methods 0.000 claims abstract description 4
- 238000002955 isolation Methods 0.000 claims abstract description 4
- 241000221638 Morchella Species 0.000 claims abstract description 3
- 241001634871 Terfezia boudieri Species 0.000 claims abstract 3
- 235000002777 Tuber melanosporum Nutrition 0.000 claims description 7
- 238000000338 in vitro Methods 0.000 claims description 7
- 235000015097 nutrients Nutrition 0.000 claims description 7
- 235000016709 nutrition Nutrition 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 5
- 244000075850 Avena orientalis Species 0.000 claims description 3
- 235000007319 Avena orientalis Nutrition 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 claims description 3
- 241000221198 Basidiomycota Species 0.000 claims description 2
- 230000000694 effects Effects 0.000 claims 8
- 241000233855 Orchidaceae Species 0.000 claims 5
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims 4
- 239000007642 baf medium Substances 0.000 claims 4
- 208000005156 Dehydration Diseases 0.000 claims 2
- 241001290329 Tricholoma terreum Species 0.000 claims 2
- 230000007613 environmental effect Effects 0.000 claims 2
- 230000009105 vegetative growth Effects 0.000 claims 2
- 101100281682 Danio rerio fsta gene Proteins 0.000 claims 1
- 241001517183 Epidendrum Species 0.000 claims 1
- 240000006383 Epidendrum nocturnum Species 0.000 claims 1
- 230000001419 dependent effect Effects 0.000 claims 1
- 150000002224 folic acids Chemical class 0.000 claims 1
- 238000004362 fungal culture Methods 0.000 claims 1
- 239000003375 plant hormone Substances 0.000 claims 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 claims 1
- 241000894007 species Species 0.000 abstract description 10
- 240000002769 Morchella esculenta Species 0.000 abstract description 5
- 235000002779 Morchella esculenta Nutrition 0.000 abstract description 4
- 241000972296 Choiromyces meandriformis Species 0.000 abstract 1
- 241000609666 Tuber aestivum Species 0.000 abstract 1
- 241000142927 Tuber magnatum Species 0.000 abstract 1
- 230000006037 cell lysis Effects 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000002093 peripheral effect Effects 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 210000001187 pylorus Anatomy 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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Abstract
Description
La presente invención proporciona un procedimiento para la obtención in vitro de micelio fúngico en cultivo puro, aislado, tanto de los cuerpos fructíferos de especies fúngicas de interés, como de secciones de raicillas de plantas micorrizadas con dichas especies fúngicas. The present invention provides a method for obtaining in vitro fungal mycelium in pure, isolated culture, both from the fruiting bodies of fungal species of interest, and from root sections of mycorrhizal plants with said fungal species.
La invención es una adición a la Patente de Invención P200930246 por “Procedimiento para la Obtención a Gran Escala de Inóculo Miceliar de Basidiomicetos”, que comprende las etapas: aislamiento del micelio fúngico de especies de hongos Ascomicetos, preferiblemente de los géneros Tuber, Terfezia, o Morchella y más preferiblemente a partir de cuerpos fructíferos de los hongos; inoculación del micelio aislado en la etapa anterior en un medio de cultivo sólido de avena modificado (MOM); purificación del micelio en medio de cultivo sólido de avena modificado (MOM); desarrollo del micelio purificado en la etapa anterior en un medio de cultivo BiotinaAneurina-Ácido Fólico (BAF) modificado y obtención del micelio fúngico purificado. The invention is an addition to the Invention Patent P200930246 for "Procedure for the Large-scale Obtaining of Mycelial Inocula of Basidiomycetes", which comprises the steps: isolation of the fungal mycelium from species of Ascomycete fungi, preferably from the genus Tuber, Terfezia, or Morchella and more preferably from fruiting bodies of fungi; inoculation of the mycelium isolated in the previous stage in a solid modified oats culture medium (MOM); mycelium purification in solid modified oatmeal culture medium (MOM); development of the purified mycelium in the previous stage in a modified BiotinAneurin-Folic Acid (BAF) culture medium and obtaining the purified fungal mycelium.
En una realización preferible de la invención, dicho hongo del género Tuber es Tuber melanosporum, y en otra realización preferible dicho hongo del género Terfezia es Terfezia arenaria. En una realización preferible más de la invención dicho medio líquido BAF modificado presenta un pH de 6,5. In a preferable embodiment of the invention, said fungus of the genus Tuber is Tuber melanosporum, and in another preferable embodiment said fungus of the genus Terfezia is Terfezia arenaria. In a further preferred embodiment of the invention said modified BAF liquid medium has a pH of 6.5.
Los resultados de los ejemplos muestran un extraordinario desarrollo y una gran cantidad de micelio obtenido. A los 5 días de cultivo se obtuvo por término medio un crecimiento de 1:103 del micelio en cultivo puro, ya dispuesto para la inoculación de las plantas a micorrizar. The results of the examples show an extraordinary development and a large amount of mycelium obtained. After 5 days of culture, a growth of 1: 103 of the mycelium in pure culture was obtained, already ready for the inoculation of the plants to be mycorrhized.
Las figuras muestran dos perspectivas diferentes del sistema utilizado para la purificación del micelio fúngico. The figures show two different perspectives of the system used for purification of the fungal mycelium.
La figura 1 corresponde a una placa de Petri de un diámetro de 9 cm (1) en cuyo fondo se coloca de forma centrada una placa (2) de 5 cm de diámetro, que se muestra con trazo discontinuo. Como resultado de esta configuración entre ambas placas se crea un espacio circular de 2 cm de ancho (3) en el que se deposita el medio de cultivo fundido. Figure 1 corresponds to a Petri dish of a diameter of 9 cm (1) at the bottom of which a plate (2) of 5 cm in diameter is placed centrally, shown in broken lines. As a result of this configuration between both plates a circular space of 2 cm wide (3) is created in which the molten culture medium is deposited.
La figura 2 muestra el espacio central (4) que queda cuando, una vez enfriado el medio, se retira la placa interpuesta. En dicho espacio vacío central es donde se sitúa el inóculo del micelio, que se desarrolla hasta alcanzar la sección periférica (3) donde se encuentra el medio para la purificación, liberándose así de la posible presencia de bacterias, las cuales quedan retenidas en la sección central de las placas, al no poder desarrollarse por ausencia de nutrientes. Figure 2 shows the central space (4) that remains when, once the medium has cooled, the interposed plate is removed. In this central empty space is where the mycelium inoculum is located, which develops until reaching the peripheral section (3) where the means for purification is found, thus freeing the possible presence of bacteria, which are retained in the section central plates, unable to develop due to lack of nutrients.
Con la intención de mostrar la presente invención de un modo ilustrativo aunque en ningún modo limitante, se aportan los siguientes ejemplos. With the intention of showing the present invention in an illustrative manner but in no way limiting, the following examples are provided.
Ejemplo 1: Preparación del inóculo miceliar de Tuber melanosporum, Terfezia arenaria y Morchella esculenta. Example 1: Preparation of the mycelial inoculum of Tuber melanosporum, Terfezia arenaria and Morchella esculenta.
Se aislaron en cámara de flujo laminar pequeñas secciones de la parte más interna del himenio del píleo de carpóforos maduros de cada especie por separado, a los que se les había eliminado previamente la pellis. Seguidamente, se inocularon dichas secciones en condiciones de máxima asepsia en cámara de flujo laminar en placas Petri de plástico de 5 cm de diámetro, previamente esterilizadas en autoclave. A estas placas Petri se les había adicionado con anterioridad medio de cultivo sólido de avena modificado (MOM) con composición/litro de medio de 0,5 g de Ca(NO3)2 4H2O; 0,2 g de PO4KH2 ; 0,1 g de SO4Mg 7H2O; 0,1g de KCl; 5,0 g de sacarosa; 3,5 g de avena en polvo; 0,1 g de extracto de levadura y 8,0 g de agar. Las placas inoculadas se mantuvieron en cámara a 20ºC hasta que el micelio se desarrolló llegando a ocupar toda la superficie de la placa. Posteriormente, el micelio obtenido en ambos casos se sometió a un proceso de purificación con objeto de eliminar la posibilidad de existencia de contaminación bacteriana, utilizando para ello placas Petri de 9 cm de diámetro en las que el medio de cultivo MOM tal cual anteriormente descrito se dispuso solamente en una sección circular periférica de 2 cm (placas de 5 cm de diámetro introducidas dentro de las placas Petri de 9 cm). En la parte central de estas placas se colocaron secciones de 1x1cm del medio sólido de crecimiento MOM conteniendo el micelio fúngico. Estas placas se mantuvieron en cámara a 20ºC hasta que el micelio se desarrolló ocupando toda la superficie de la placa. Small sections of the innermost part of the hymenium of the mature carpal pylorus of each species were isolated in a laminar flow chamber, which had previously been removed from the pellis. Then, said sections were inoculated under conditions of maximum aseptic laminar flow chamber in plastic Petri dishes 5 cm in diameter, previously autoclaved. These Petri dishes had previously been added modified solid oatmeal culture medium (MOM) with a composition / liter of 0.5 g Ca (NO3) 2 4H2O medium; 0.2 g of PO4KH2; 0.1 g of SO4Mg 7H2O; 0.1g of KCl; 5.0 g sucrose; 3.5 g of oatmeal powder; 0.1 g of yeast extract and 8.0 g of agar. The inoculated plates were kept in a chamber at 20 ° C until the mycelium developed to occupy the entire surface of the plate. Subsequently, the mycelium obtained in both cases was subjected to a purification process in order to eliminate the possibility of bacterial contamination, using 9 cm diameter Petri dishes in which the MOM culture medium as described above is arranged only in a peripheral circular section of 2 cm (5 cm diameter plates inserted into the 9 cm Petri dishes). In the central part of these plates, 1x1cm sections of the solid MOM growth medium containing the fungal mycelium were placed. These plates were kept in a chamber at 20 ° C until the mycelium was developed occupying the entire surface of the plate.
Ejemplo 2: Cultivo del inóculo miceliar Example 2: Culture of mycelial inoculum
Una vez purificado el micelio de cada especie según el Ejemplo 1 se transfirió una pequeña porción para su desarrollo a gran escala a medio de cultivo líquido BAF 5 modificado. En todos los experimentos se partió de un inóculo micelial de tamaño similar con un peso medio de unos 10 mg, que se incubó en matraces erlenmeyer conteniendo medio de cultivo líquido BAF modificado con una composición/litro de medio: 0,1 g de Cl2Ca; 0,5 g de PO4KH2; 0,5 g de SO4Mg 7H2O; 0,005 g de SO4Mn; 0,001 g de SO4Zn; 0,01 g de Cl3Fe 7H2O; 2,0 g de peptona; 0,0005 g de tiamina; Once the mycelium of each species was purified according to Example 1, a small portion was transferred for large-scale development to modified BAF 5 liquid culture medium. In all experiments, a mycelial inoculum of similar size with an average weight of about 10 mg was started, which was incubated in erlenmeyer flasks containing modified BAF liquid culture medium with a composition / liter of medium: 0.1 g of Cl2Ca; 0.5 g of PO4KH2; 0.5 g of SO4Mg 7H2O; 0.005 g of SO4Mn; 0.001 g of SO4Zn; 0.01 g of Cl3Fe 7H2O; 2.0 g peptone; 0.0005 g of thiamine;
10 0,00001 g de biotina; 0,05 g de inositol; 0,0001 g de ácido fólico; 5,0-20,0 g de glucosa, y 0,2 g de extracto de levadura; el cultivo se mantuvo bajo condiciones controladas: 25 ºC, oscuridad, pH 6,5 y agitación constante a 120 rpm durante todos los días del cultivo ininterrumpidamente. 10 0.00001 g of biotin; 0.05 g of inositol; 0.0001 g of folic acid; 5.0-20.0 g of glucose, and 0.2 g of yeast extract; The culture was maintained under controlled conditions: 25 ° C, darkness, pH 6.5 and constant agitation at 120 rpm during all days of the crop continuously.
Los resultados del crecimiento miceliar en medio nutritivo líquido BAF de la especie Tuber melanosporum durante catorce días sin nuevos aportes de medio nutritivo, se muestran en la tabla I. El crecimiento se mantiene hasta el día 14 en que, a causa del agotamiento de los nutrientes del medio de cultivo, se observa un cierto descenso del The results of the mycelial growth in liquid nutritional medium BAF of the species Tuber melanosporum for fourteen days without new contributions of nutritional medium, are shown in Table I. The growth is maintained until day 14 when, due to the depletion of nutrients of the culture medium, a certain decrease in the
20 mismo, que evidencia el inicio de procesos de lisis celular. Por ello, después de estos 14 días se requirió un nuevo aporte de medio nutritivo para la reanudación del crecimiento del micelio. 20 which evidences the beginning of cell lysis processes. Therefore, after these 14 days a new supply of nutrient media was required for the resumption of mycelium growth.
Tabla I Table I
- Peso medio del micelio (gr) Average weight of mycelium (gr)
- Especie Species
- día 0 día 3 día 5 día 8 día 12 día 14 day 0 day 3 day 5 day 8 day 12 day 14
- T.melanosporum T. melanosporum
- 0,0081 5,1681 7,9722 10,9754 12,1732 11,7312 0.0081 5,1681 7.9722 10,9754 12.1732 11.7312
25 De estos datos se deduce que a los cinco días el crecimiento del micelio en cultivo puro fue de 1:103, lo que se corresponde con una producción a gran escala. 25 It follows from these data that after five days the growth of mycelium in pure culture was 1: 103, which corresponds to a large-scale production.
Ejemplo 4: Obtención de micelio de Terfezia arenaria en cultivo puro Example 4: Obtaining mycelium of Terfezia arenaria in pure culture
30 El crecimiento miceliar en medio nutritivo líquido BAF de la especie Terfezia arenaria durante catorce días sin nuevos aportes de medio nutritivo, se muestra en la tabla II. 30 The mycelial growth in the BAF liquid nutrient medium of the Terfezia arenaria species for fourteen days without new contributions of the nutrient medium, is shown in Table II.
En este ejemplo también se mantiene el crecimiento constante hasta el día 14 en que se observa un cierto descenso del mismo, que evidencia el inicio de los procesos de lisis celular. A partir de los 14 días se requirió un nuevo aporte de medio nutritivo para la reanudación del crecimiento del micelio. In this example, constant growth is also maintained until day 14 when a certain decrease is observed, which evidences the start of cell lysis processes. After 14 days, a new supply of nutrient medium was required for the resumption of mycelium growth.
Tabla II Table II
- Peso medio del micelio (gr) Average weight of mycelium (gr)
- Especie Species
- día 0 día 3 día 5 día 8 día 12 día 14 day 0 day 3 day 5 day 8 day 12 day 14
- T.arenaria T. arenaria
- 0,0111 4,9051 7,6351 9,2431 10,3065 10,0271 0.0111 4,9051 7.6351 9.2431 10,3065 10,0271
Al igual que en la especie anterior, de los datos expuestos en la tabla se deduce que a los cinco días el crecimiento fue de 1:7x102, lo que indica también un extraordinario As in the previous species, from the data presented in the table it follows that after five days the growth was 1: 7x102, which also indicates an extraordinary
10 desarrollo, si bien, un poco menor que en el ejemplo anterior. 10 development, although a little less than in the previous example.
Ejemplo 5: Obtención de micelio de Morchella esculenta en cultivo puro Example 5: Obtaining mycelium of Morchella esculenta in pure culture
El crecimiento miceliar en medio nutritivo líquido BAF de la especie Morchella esculenta. El crecimiento se mantiene, asimismo, hasta el día 14 en que se observa un The mycelial growth in liquid nutritional medium BAF of the species Morchella esculenta. The growth is also maintained until the 14th day when a
15 cierto descenso del mismo, que evidencia el inicio de procesos de lisis celular. Después de estos 14 días se requirió, igualmente, un nuevo aporte de medio nutritivo para la reanudación del crecimiento del micelio. 15 certain decrease in it, which evidences the beginning of cell lysis processes. After these 14 days, a new supply of nutritional medium was also required for the resumption of mycelium growth.
- Peso medio del micelio (gr) Average weight of mycelium (gr)
- Especie Species
- día 0 día 3 día 5 día 8 día 12 día 14 day 0 day 3 day 5 day 8 day 12 day 14
- M.esculenta M.
- 0,0078 4,6273 7,6895 10,7395 12,6912 12,0161 0.0078 4.6273 7.6895 10,7395 12.6912 12,0161
20 De los datos expuestos en la tabla se deduce que a los cinco días el crecimiento fue de 1:103, lo que se corresponde igualmente con una producción a gran escala similar a la del Ejemplo 3. 20 From the data presented in the table it follows that after five days the growth was 1: 103, which also corresponds to a large-scale production similar to that of Example 3.
Claims (7)
- 1. one.
- Adición a la Patente de Invención P200930246 por “Procedimiento para la Obtención a Gran Escala de Inóculo Miceliar de Basidiomicetos”, que comprende las etapas: Addition to the Invention Patent P200930246 for "Procedure for the Large-scale Obtaining of Mycelial Inocula of Basidiomycetes", which comprises the steps:
- 2. 2.
- Adición a la Patente de Invención P200930246 según la reivindicación 1, caracterizada por que dicho aislamiento del micelio fúngico de la etapa a) se realiza a partir de cuerpos fructíferos de hongos. Addition to the Patent of Invention P200930246 according to claim 1, characterized in that said isolation of the fungal mycelium of step a) is carried out from fruiting fungal bodies.
- 3. 3.
- Adición a la Patente de Invención P200930246 según la reivindicación 2, caracterizado por que el hongo de ascomicetos a partir del cual se obtiene el micelio es de los géneros Tuber, Terfezia, o Morchella. Addition to the invention patent P200930246 according to claim 2, characterized in that the ascomycete fungus from which the mycelium is obtained is of the genera Tuber, Terfezia, or Morchella.
- 4. Four.
- Adición a la Patente de Invención P200930246 según la reivindicación 3, caracterizado por que dicho hongo del género Tuber es Tuber melanosporum. Addition to the invention patent P200930246 according to claim 3, characterized in that said fungus of the genus Tuber is Tuber melanosporum.
- 5. 5.
- Adición a la Patente de Invención P200930246 según la reivindicación 3, caracterizado por que dicho hongo del género Terfezia es Terfezia arenaria. Addition to the invention patent P200930246 according to claim 3, characterized in that said fungus of the genus Terfezia is Terfezia arenaria.
- 6. 6.
- Adición a la Patente de Invención P200930246 según la reivindicación 1, caracterizado por que dicho medio líquido BAF modificado de la etapa d) presenta un pH de 6,5. Addition to the invention patent P200930246 according to claim 1, characterized in that said modified BAF liquid medium of step d) has a pH of 6.5.
- Categoría Category
- 56 Documentos citados Reivindicaciones afectadas 56 Documents cited Claims Affected
- A TO
- VÁZQUEZ-GARCÍA, A . et al . I nfluencia del pH en el cr ecimiento d e q uince ce pas de h ongos ectomicorrizógenos. Anales del Instituto de Biología, Universidad Autónoma de México. Serie Botánica. 2002. Volumen 73. Número 1. Páginas 1-15. ISSN 0185-254X. 1-6 VÁZQUEZ-GARCÍA, A. et al. The influence of pH on the growth of the ectomycorrhizal fungi. Annals of the Institute of Biology, Autonomous University of Mexico. Botanical Series 2002. Volume 73. Number 1. Pages 1-15. ISSN 0185-254X. 1-6
- A TO
- KIBAR, B. et al. Nutritional and environmental requirements for vegetative growth of edible 1-6 KIBAR, B. et al. Nutritional and environmental requirements for vegetative growth of edible 1-6
- ectomycorrhizal mushroom Tricholoma terreum. Zemdirbysté=Agriculture. 2011. Volumen 98. Nº 4. Páginas 409-414. ISSN 1392-3196. ectomycorrhizal mushroom Tricholoma terreum. Zemdirbysté = Agriculture. 2011. Volume 98. No. 4. Pages 409-414. ISSN 1392-3196.
- A TO
- ZETTLER, L. W. et al . Conservation-driven pr opagation of an ep iphytic orchid ( Epidendrum nocturnum) with a mycorrhizal fungus. HortScience. Febrero, 2007. Vol. 2, Nº 1, páginas 135-139. ISSN 0018-5345. 1-6 ZETTLER, L. W. et al. Conservation-driven pr opagation of an ep iphytic orchid (Epidendrum nocturnum) with a mycorrhizal fungus. HortScience February 2007. Vol. 2, No. 1, pages 135-139. ISSN 0018-5345. 1-6
- A TO
- KAMAL, S. et al. Effect of nu trient so urces and plant h ormones on m ycelial m orphology of t he black perigord truffle Tuber melanosporum. Proceedings of the 7th International Conference on Mushroom Biology and Mushroom Products. 2011. Páginas 509-515. 1-6 KAMAL, S. et al. Effect of nu trient so urces and plant h ormones on m ycelial m orphology of t he black perigord truffle Tuber melanosporum. Proceedings of the 7th International Conference on Mushroom Biology and Mushroom Products. 2011. Pages 509-515. 1-6
- A TO
- NAVARRO-RÓDENAS, A . e t a l. Effect o f w ater st ress on i n v itro m ycelium cu ltures of t wo mycorrhizal desert truffles. Mycorrhiza. Mayo, 2011. Volumen 21, Nº 4. Páginas 247-253. ISSN 1432-1890. doi:10.1007/s00572-010-0329-z 1-6 NAVARRO-RÓDENAS, A. e t a l. Effect o f w ater st ress on i n v itro m ycelium cu ltures of t wo mycorrhizal desert truffles. Mycorrhiza May 2011. Volume 21, No. 4. Pages 247-253. ISSN 1432-1890. doi: 10.1007 / s00572-010-0329-z 1-6
- Categoría de los documentos citados X: de particular relevancia Y: de particular relevancia combinado con otro/s de la misma categoría A: refleja el estado de la técnica O: referido a divulgación no escrita P: publicado entre la fecha de prioridad y la de presentación de la solicitud E: documento anterior, pero publicado después de la fecha de presentación de la solicitud Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application
- El presente informe ha sido realizado • para todas las reivindicaciones □ para las reivindicaciones nº: This report has been prepared • for all claims □ for claims no:
- Fecha de realización del informe 14.09.2012 Date of realization of the report 14.09.2012
- Examinador E. M. Ulloa Calvo Página 1/4 Examiner E. M. Ulloa Calvo Page 1/4
- Novedad (Art. 6.1 LP 11/1986) Novelty (Art. 6.1 LP 11/1986)
- Reivindicaciones Reivindicaciones 1-6 SI NO Claims Claims 1-6 IF NOT
- Actividad inventiva (Art. 8.1 LP11/1986) Inventive activity (Art. 8.1 LP11 / 1986)
- Reivindicaciones Reivindicaciones 1-6 SI NO Claims Claims 1-6 IF NOT
- Documento Document
- Número Publicación o Identificación Fecha Publicación Publication or Identification Number publication date
- D01 D01
- VÁZQUEZ-GARCÍA, A. et al. Influencia del pH en el crecimiento de quince cepas de hongos ectomicorrizógenos. 2002 VÁZQUEZ-GARCÍA, A. et al. Influence of pH on the growth of fifteen strains of ectomycorrhizal fungi. 2002
- D02 D02
- KIBAR, B. et al. Nutritional and environmental requirements for vegetative growth of ed ible ect omycorrhizal mushroom Tricholoma terreum. 2011 KIBAR, B. et al. Nutritional and environmental requirements for vegetative growth of edible ect omycorrhizal mushroom Tricholoma terreum. 2011
- D03 D03
- ZETTLER, L .W. e t a l. Conservation-driven pr opagation of a n epiphytic orchid ( Epidendrum n octurnum) with a m ycorrhizal fungus. 2007 ZETTLER, L .W. e t a l. Conservation-driven pr opagation of a n epiphytic orchid (Epidendrum n octurnum) with a m ycorrhizal fungus. 2007
- D04 D04
- KAMAL, S. et al. Effect of nutrient sources and plant hormones on mycelial m orphology of t he black perigord truffle T uber melanosporum. 2011 KAMAL, S. et al. Effect of nutrient sources and plant hormones on mycelial m orphology of t he black perigord truffle T uber melanosporum. 2011
- D05 D05
- NAVARRO-RÓDENAS, A. et al. Effect of water stress on in vitro mycelium cultures of two mycorrhizal desert truffles.. 2011 NAVARRO-RÓDENAS, A. et al. Effect of water stress on in vitro mycelium cultures of two mycorrhizal desert truffles .. 2011
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108522139A (en) * | 2018-04-25 | 2018-09-14 | 赵金亮 | A kind of efficient implantation methods of brooder hickory chick |
| EP4009778A4 (en) * | 2019-08-09 | 2023-09-27 | Carolina Truffiéres LLC | Methods and compositions for the production of ectomycorrhizal mycelia and methods of use thereof |
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| US20220104523A1 (en) * | 2019-01-09 | 2022-04-07 | Mycotechnology, Inc. | Food and beverage products comprising ascomycetes |
| PT3811773T (en) | 2019-10-22 | 2024-07-09 | Univ Evora | Culture media and process for improved isolation and maintenance of terfezia spp. mycelium cultures |
| CN115715146A (en) | 2020-06-25 | 2023-02-24 | 麦可科技有限公司 | Sweet protein from truffle |
| CN113774091B (en) * | 2021-09-23 | 2023-08-18 | 山东安为先生物科技有限公司 | Stable water-soluble nano-selenium and preparation method thereof |
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| ES2332031B2 (en) * | 2009-06-02 | 2010-05-31 | Universidad Politecnica De Madrid | PROCEDURE FOR OBTAINING A LARGE SCALE OF MICELLIUM INOCULATE OF BASIDIOMICETS. |
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Non-Patent Citations (5)
| Title |
|---|
| KAMAL, S. et al. Effect of nutrient sources and plant hormones on mycelial morphology of the black perigord truffle Tuber melanosporum. Proceedings of the 7th International Conference on Mushroom Biology and Mushroom Products. 2011. Páginas 509-515. * |
| KIBAR, B. et al. Nutritional and environmental requirements for vegetative growth of edible ectomycorrhizal mushroom Tricholma terreum. Zemdirbysté=Agriculture. 2011. Volumen 98. Nº 4. Páginas 409-414. ISSN 1392-3196. * |
| NAVARRO-RÓDENAS, A. et al. Effect of water stress on in vitro mycelium cultures of two mycorrhizal desert truffles. Mycorrhiza. Mayo, 2011. Volumen 21, Nº 4. Páginas 247-253. ISSN 1432-1890. doi:10.1007/s00572-010-0329-z . * |
| VÁZQUEZ-GARCÍA, A. et al. Influencia del pH en el crecimiento de quince cepas de hongos ectomicorrizógenos. Anales del Instituto de Biología, Universidad Autónoma de México. Serie Botánica. 2002. Volumen 73. Número 1. Páginas 1-15. ISSN 0185-254X. * |
| ZETTLER, L.W. et al. Conservation-driven propagation of an epiphytic orchid (Epidendrum nocturnum) with a mycorrhizal fungus. HortScience. Febrero, 2007. Vol. 2, Nº 1, páginas 135- 139. ISSN 0018-5345. * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108522139A (en) * | 2018-04-25 | 2018-09-14 | 赵金亮 | A kind of efficient implantation methods of brooder hickory chick |
| EP4009778A4 (en) * | 2019-08-09 | 2023-09-27 | Carolina Truffiéres LLC | Methods and compositions for the production of ectomycorrhizal mycelia and methods of use thereof |
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| WO2014020215A1 (en) | 2014-02-06 |
| ES2387666B2 (en) | 2013-01-24 |
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