WO2003017754A1 - Modified taxus-type plant species and the production method therefor - Google Patents
Modified taxus-type plant species and the production method therefor Download PDFInfo
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- WO2003017754A1 WO2003017754A1 PCT/ES2002/000393 ES0200393W WO03017754A1 WO 2003017754 A1 WO2003017754 A1 WO 2003017754A1 ES 0200393 W ES0200393 W ES 0200393W WO 03017754 A1 WO03017754 A1 WO 03017754A1
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- concentration
- weight
- taxanes
- plant species
- leaves
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- 241000196324 Embryophyta Species 0.000 title claims abstract description 53
- 238000004519 manufacturing process Methods 0.000 title description 2
- 229940123237 Taxane Drugs 0.000 claims abstract description 41
- 241001116500 Taxus Species 0.000 claims description 36
- 229930000044 secondary metabolite Natural products 0.000 claims description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 11
- 241000894007 species Species 0.000 claims description 11
- 235000013311 vegetables Nutrition 0.000 claims description 9
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 abstract description 8
- 229910002092 carbon dioxide Inorganic materials 0.000 abstract description 4
- 239000001569 carbon dioxide Substances 0.000 abstract description 3
- 238000002715 modification method Methods 0.000 abstract 1
- RCINICONZNJXQF-XAZOAEDWSA-N taxol® Chemical compound O([C@@H]1[C@@]2(CC(C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3(C21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-XAZOAEDWSA-N 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 239000002028 Biomass Substances 0.000 description 4
- 229930012538 Paclitaxel Natural products 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 229960001592 paclitaxel Drugs 0.000 description 4
- -1 diterpene amide taxane Chemical class 0.000 description 3
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 241000202349 Taxus brevifolia Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000024053 secondary metabolic process Effects 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000719 anti-leukaemic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 229930004069 diterpene Natural products 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 208000005026 persistent vegetative state Diseases 0.000 description 1
- 235000010204 pine bark Nutrition 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/02—Treatment of plants with carbon dioxide
Definitions
- the present invention relates to a modified plant species of the genus Taxus and the method of obtaining it.
- the present invention also relates to the use of carbon dioxide (C0 2 ) to increase the concentration of natural products from secondary metabolism in a plant species.
- the modified plant species has levels in taxanes (especially paclitaxel, also known as taxol®) higher than those found in the unmodified species.
- Carbon fertilizer in greenhouses is a well-known technique that is used to increase biomass in trees and, in agriculture, to increase fruit quality.
- Taxol® the compound known as paclitaxel
- Taxol® is the only diterpene amide taxane that has antitumor and antileukemic properties. Therefore, it is of great interest to obtain this therapeutic chemical agent for the clinical treatment of patients presenting with breast cancer.
- the taxol® exists only in small quantities, of the order of 0.01% inside the bark, in the species of Tejo ⁇ Taxus).
- taxol® could not be synthesized and even now, the most economical source of this compound still comes from the Pacific Yew (Taxus brevifolia). For this reason, large areas of Pacific Tejo forests have been destroyed in the northeastern United States in order to obtain this anticancer drug.
- the paclitaxel compound is, therefore, a secondary metabolite of great interest for its therapeutic qualities in the treatment of cancer and, therefore, there is a need to increase its extraction without this being a detriment to nature.
- the present invention relates to the method of obtaining said modified plant species.
- the present invention relates to a modified plant species of the genus Taxus that, surprisingly, comprises a concentration of taxanes higher than that presented by a plant species of the genus Taxus unmodified.
- plant species means any of the vegetative states that plants present in their natural state or when they are cultivated in vivo in greenhouses, excluding intermediate growth or propagation states used in this definition. In vitro culture techniques.
- Modified plant species of the genus Taxus means any plant species of the genus Taxus that has been subjected to the conditions of the process of the invention.
- the invention relates to a modified plant species of the genus Taxus having a bark with a concentration by weight of taxanes greater than the concentration by weight of said compounds in the bark of a plant species of the genus Taxus unmodified.
- the concentration by weight of taxanes in the bark is equal to or greater, by more than 50%, to the concentration by weight of taxanes in the bark of an unmodified plant species of the genus Taxus.
- the concentration by weight of taxanes in the bark is equal to or greater than, by more than 100%, the concentration by weight of taxanes in the bark of an unmodified plant species of the genus
- the invention also relates to a species modified vegetable of the genus Taxus that has leaves with a concentration by weight of taxanes greater than the concentration by weight of said compound in the leaves of a plant species of the genus Taxus unmodified.
- the concentration by weight of taxanes in the leaves is equal to or greater than, by more than 20%, the concentration by weight of taxanes in the leaves of an unmodified plant species of the genus Taxus.
- the concentration by weight of taxanes in the leaves is equal or greater, by more than 25%, to the concentration by weight of taxanes in the leaves of an unmodified plant species of the genus Taxus.
- the present invention also relates to the process for obtaining the modified plant species of the genus Taxus.
- the dose of C0 2 ranges between 500 and 700 ppm and the water regime, preferably, is 100%.
- secondary metabolite refers to any natural product from the secondary metabolism of a plant species.
- the present invention also relates to the use of C0 2 to increase the concentration of secondary metabolites in a plant species.
- the use C0 2 in the present invention does not cause an exclusive increase in biomass but, in addition, there is an increase in the concentration of secondary metabolites, some with therapeutic interest such as paclitaxel.
- the invention also relates to the use of C0 2 to, surprisingly, increase the concentration by weight of secondary metabolites in the leaves, bark, stems, branches, flowers and roots of a
- C0 2 causes an increase in the concentration by weight of secondary metabolites in the same or higher cortex, by more than one
- the use of C0 2 causes an increase in the weight concentration of metabolites
- the invention also relates to the use 30 of C0 2 to advantageously increase the concentration by weight of secondary metabolites in the leaves of a plant species.
- C0 2 results in an increase in the metabolite concentration by weight Secondary in the leaves equal or higher, by more than 20%, to the concentration of secondary metabolites in the leaves before submitting them to the supply of C0 2 .
- the use of C0 2 results in an increase in the concentration by weight of secondary metabolites in the leaves equal or greater, by more than 25%, to the concentration of secondary metabolites in the leaves before subjecting them to the supply of C0 2 .
- the present invention also relates to the use of C0 2 in a dose larger than 350 ppm, preferably a dose between 500 and 700 ppm, to increase the concentration of secondary metabolites in a plant species.
- Example 1 Obtaining a modified plant species of the genus Taxus.
- Yews are irrigated by spraying twice at day with a nutrient solution comprising the nutrients: P 2 ⁇ 5 : K 2 0, in a ratio 1: 0.5: 1.5 and pH 6.5.
- C0 2 was supplied at various levels (350, 500 and 700 ppm), and at each level of C0 2 different water regimes (50 and 100%, 400 and 800 m-) were supplied. year "1 respectively). Thus the yews were treated in six different conditions.
- the application of C0 2 was carried out using conventional polyethylene pipes.
- the source of C0 2 was a liquid C0 2 tank equipped with an evaporator to pass the C0 2 to the gas phase.
- the dosage of C0 2 was carried out by means of a system composed of an infrared analyzer, an automaton and a series of solenoid valves that injected C0 2 depending on the orders of the automaton.
- Example 2 Effects of enrichment with C0 2 on yew.
- the procedure followed to analyze the levels of taxanes comprises the following stages:
- the carbonic fertilizer does not promote 'significant differences in the relative growth rate (RGR) of all yew plant or its fractions.
- the tree shows a positive tendency towards growth in the presence of higher levels of C0 2 during the two years.
- plants grown at 700 ppm contain 28.8% more taxanes in 400 mm-years "" 1 and 49.36% more taxanes in 800 mm-years "1 in the leaves compared to those submitted to 350 ppm
- FIGS 1 and 2 Relative growth rate (RGR) of dry leaf weight at 400 and 800 mm-year "1 , respectively.
- FIGS 3 and 4 Relative growth rate (RGR) of the dry weight of stems at 400 and 800 mm-year “1 , respectively.
- FIGS 5 and 6 Relative growth rate (RGR) of the dry root weight at 400 and 800 mm-year “1 , respectively.
- Figures 7 and 8 Relative growth rate (RGR) of the total dry weight at 400 and 800 mm -year "1 , respectively.
- Figure 9 Distribution of dry weight of leaves grown at three doses of C0 2 and two of water. The values are the average of five plants ⁇ standard error.
- Figure 10 Distribution of dry weight of roots grown at three doses of C0 2 and two of water. The values are the average of five plants ⁇ standard error.
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biodiversity & Conservation Biology (AREA)
- Botany (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Environmental Sciences (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Epoxy Compounds (AREA)
Abstract
The invention relates to a Taxus-type plant species which is modified in the presence of carbon dioxide and which has a weight concentration of taxanes that is greater than that present in a non-modified Taxus-type plant species. The modification method consists in administering a dose of carbon dioxide which is greater than 350 ppm and subjecting the plants to a moisture regime of between 50 and 100 %.
Description
ESPECIE VEGETAL MODIFICADA DEL GENERO TAXUS, PROCEDIMIENTOMODIFIED VEGETABLE SPECIES OF THE GENRE TAXUS, PROCEDURE
PARA SU OBTENCIÓN Y UTILIZACIÓN DE C02 PARA INCREMENTAR LAFOR ITS OBTAINING AND USE OF C0 2 TO INCREASE THE
CONCENTRACIÓN DE METABOLITOS SECUNDARIOS EN UNA ESPECIECONCENTRATION OF SECONDARY METABOLITES IN A SPECIES
VEGETAL .VEGETABLE
CAMPO DE LA INVENCIÓNFIELD OF THE INVENTION
La presente invención se refiere a una especie vegetal modificada del género Taxus y al procedimiento de obtención de la misma.The present invention relates to a modified plant species of the genus Taxus and the method of obtaining it.
La presente invención también se refiere a la utilización del dióxido de carbono (C02) para aumentar la concentración de productos naturales procedentes del metabolismo secundario en una especie vegetal.The present invention also relates to the use of carbon dioxide (C0 2 ) to increase the concentration of natural products from secondary metabolism in a plant species.
La especie vegetal modificada, según la presente invención, presenta unos niveles en taxanos (especialmente paclitaxel, también conocido como taxol®) superiores a los que se encuentran en la especie no modificada.The modified plant species, according to the present invention, has levels in taxanes (especially paclitaxel, also known as taxol®) higher than those found in the unmodified species.
ANTECEDENTES DE LA INVENCIÓNBACKGROUND OF THE INVENTION
El abono carbónico en invernaderos es una técnica bastante conocida que se utiliza con el fin de aumentar la biomasa en árboles y, en agricultura, para aumentar la calidad del fruto.Carbon fertilizer in greenhouses is a well-known technique that is used to increase biomass in trees and, in agriculture, to increase fruit quality.
Por otro lado, el compuesto conocido como paclitaxel (taxol®) es el único taxano diterpeno amida que posee propiedades antitumorales y antileucémicas. Por lo tanto, es de gran interés la obtención de este agente químico terapéutico para el tratamiento clínico de pacientes que presentan cáncer de mama. Sin embargo, el
taxol® existe sólo en pequeñas cantidades, del orden de 0,01% en el interior de la corteza, en las especies de Tejo { Taxus) . Hasta no hace muchos años, el taxol® no podia sintetizarse e incluso ahora, la fuente más económica de este compuesto procede todavía del Tejo del Pacifico { Taxus brevifolia ) . Por este motivo, se han destruido grandes áreas de bosques de Tejo del Pacifico en el nordeste de Estados Unidos con el fin de obtener éste fármaco anticancerigeno.On the other hand, the compound known as paclitaxel (taxol®) is the only diterpene amide taxane that has antitumor and antileukemic properties. Therefore, it is of great interest to obtain this therapeutic chemical agent for the clinical treatment of patients presenting with breast cancer. However the taxol® exists only in small quantities, of the order of 0.01% inside the bark, in the species of Tejo {Taxus). Until not many years ago, taxol® could not be synthesized and even now, the most economical source of this compound still comes from the Pacific Yew (Taxus brevifolia). For this reason, large areas of Pacific Tejo forests have been destroyed in the northeastern United States in order to obtain this anticancer drug.
El compuesto paclitaxel es, pues, un metabolito secundario de gran interés por sus cualidades terapéuticas en el tratamiento del cáncer y, por lo tanto, existe una necesidad de aumentar su extracción sin que ello represente un perjuicio para la naturaleza.The paclitaxel compound is, therefore, a secondary metabolite of great interest for its therapeutic qualities in the treatment of cancer and, therefore, there is a need to increase its extraction without this being a detriment to nature.
Es un objeto de la presente invención la obtención de una especie vegetal modificada del género Taxus que se caracteriza por presentar una concentración de taxanos y, en particular de taxol®, superiores a la que presenta la especie vegetal sin tratar.It is an object of the present invention to obtain a modified plant species of the genus Taxus that is characterized by a concentration of taxanes and, in particular of taxol®, higher than that presented by the untreated plant species.
De la misma manera, la presente invención se refiere al procedimiento de obtención de dicha especie vegetal modificada.In the same way, the present invention relates to the method of obtaining said modified plant species.
Es otro objetivo de la invención aumentar los niveles de metabolitos secundarios en especies vegetales mediante la utilización de C02.It is another objective of the invention to increase the levels of secondary metabolites in plant species by using C0 2 .
DESCRIPCIÓN DE LA INVENCIÓNDESCRIPTION OF THE INVENTION
La presente invención se refiere a una especie vegetal modificada del género Taxus que, sorprendentemente, comprende una concentración de taxanos
superior a la que presenta una especie vegetal del género Taxus sin modificar.The present invention relates to a modified plant species of the genus Taxus that, surprisingly, comprises a concentration of taxanes higher than that presented by a plant species of the genus Taxus unmodified.
A efectos de la presente invención, por "especie vegetal" se entiende cualquiera de los estados vegetativos que presentan las plantas en su estado natural o cuando son cultivadas in vivo en invernaderos, quedando excluida de esta definición los estados intermedios de crecimiento o propagación empleados en las técnicas de cultivo in vitro .For the purposes of the present invention, "plant species" means any of the vegetative states that plants present in their natural state or when they are cultivated in vivo in greenhouses, excluding intermediate growth or propagation states used in this definition. In vitro culture techniques.
Por "especie vegetal modificada del género Taxus" se entenderá cualquier especie vegetal del género Taxus que haya sido sometida a las condiciones del procedimiento de la invención."Modified plant species of the genus Taxus" means any plant species of the genus Taxus that has been subjected to the conditions of the process of the invention.
Más concretamente, la invención se refiere a una especie vegetal modificada del género Taxus que tiene una corteza con una concentración en peso de taxanos superior a la concentración en peso de dichos compuestos en la corteza de una especie vegetal del género Taxus no modificada .More specifically, the invention relates to a modified plant species of the genus Taxus having a bark with a concentration by weight of taxanes greater than the concentration by weight of said compounds in the bark of a plant species of the genus Taxus unmodified.
Preferiblemente, la concentración en peso de taxanos en la corteza es igual o superior, en más de un 50%, a la concentración en peso de taxanos en la corteza de una especie vegetal no modificada del género Taxus .Preferably, the concentration by weight of taxanes in the bark is equal to or greater, by more than 50%, to the concentration by weight of taxanes in the bark of an unmodified plant species of the genus Taxus.
Todavía más preferiblemente, la concentración en peso de taxanos en la corteza es igual o superior, en más de un 100%, a la concentración en peso de taxanos en la corteza de una especie vegetal no modificada del generoEven more preferably, the concentration by weight of taxanes in the bark is equal to or greater than, by more than 100%, the concentration by weight of taxanes in the bark of an unmodified plant species of the genus
Taxus .Taxus
La invención también se refiere a una especie
vegetal modificada del género Taxus que tiene hojas con una concentración en peso de taxanos superior a la concentración en peso de dicho compuesto en las hojas de una especie vegetal del género Taxus no modificada.The invention also relates to a species modified vegetable of the genus Taxus that has leaves with a concentration by weight of taxanes greater than the concentration by weight of said compound in the leaves of a plant species of the genus Taxus unmodified.
Preferiblemente, la concentración en peso de taxanos en las hojas es igual o superior, en más de un 20%, a la concentración en peso de taxanos en las hojas de una especie vegetal no modificada del género Taxus.Preferably, the concentration by weight of taxanes in the leaves is equal to or greater than, by more than 20%, the concentration by weight of taxanes in the leaves of an unmodified plant species of the genus Taxus.
Todavía más preferiblemente, la concentración en peso de taxanos en las hojas es igual o superior, en más de un 25%, a la concentración en peso de taxanos en las hojas de una especie vegetal no modificada del género Taxus .Even more preferably, the concentration by weight of taxanes in the leaves is equal or greater, by more than 25%, to the concentration by weight of taxanes in the leaves of an unmodified plant species of the genus Taxus.
La presente invención también se refiere al procedimiento para la obtención de la especie vegetal modificada del género Taxus .The present invention also relates to the process for obtaining the modified plant species of the genus Taxus.
Dicho procedimiento se caracteriza por las siguientes condiciones experimentales:Said procedure is characterized by the following experimental conditions:
- Régimen hidrico del 50-100% (400-800 mm-año-1) . - Dosis de C02 superior 350.- Water regime of 50-100% (400-800 mm-year -1 ). - Dose of C0 2 upper 350.
Preferiblemente la dosis de C02 oscila entre 500 y 700 ppm y el régimen hidrico, preferiblemente, es del 100%.Preferably the dose of C0 2 ranges between 500 and 700 ppm and the water regime, preferably, is 100%.
A efectos de la presente solicitud, el término "metabolito secundario" se refiere a cualquier producto natural procedente del metabolismo secundario de una especie vegetal.
La presente invención también se refiere a la utilización de C02 para aumentar la concentración de metabolitos secundarios en una especie vegetal.For the purposes of the present application, the term "secondary metabolite" refers to any natural product from the secondary metabolism of a plant species. The present invention also relates to the use of C0 2 to increase the concentration of secondary metabolites in a plant species.
5 Ventajosamente, la utilización C02 en la presente invención no provoca un aumento exclusivo de biomasa sino que, además, se produce un incremento en la concentración de metabolitos secundarios, algunos con interés terapéutico como el paclitaxel .Advantageously, the use C0 2 in the present invention does not cause an exclusive increase in biomass but, in addition, there is an increase in the concentration of secondary metabolites, some with therapeutic interest such as paclitaxel.
1010
Además, la invención también se refiere a la utilización de C02 para, sorprendentemente, aumentar la concentración en peso de metabolitos secundarios en las hojas, corteza, tallos, ramas, flores y raices de unaIn addition, the invention also relates to the use of C0 2 to, surprisingly, increase the concentration by weight of secondary metabolites in the leaves, bark, stems, branches, flowers and roots of a
15 especie vegetal.15 plant species.
Más particularmente, la utilización de C02 provoca un aumento de la concentración en peso de metabolitos secundarios en la corteza igual o superior, en más de unMore particularly, the use of C0 2 causes an increase in the concentration by weight of secondary metabolites in the same or higher cortex, by more than one
20 50%, a la concentración de metabolitos secundarios en la corteza antes de someterlas al suministro de C02.20 50%, to the concentration of secondary metabolites in the cortex before submitting them to the supply of C0 2 .
Preferiblemente, la utilización de C02 provoca un aumento de la concentración en peso de metabolitosPreferably, the use of C0 2 causes an increase in the weight concentration of metabolites
25 secundarios en la corteza igual o superior, en más de un25 secondary in the same or superior cortex, in more than one
100%, a la concentración de metabolitos secundarios en la corteza antes de someterlas al suministro de C02.100%, at the concentration of secondary metabolites in the cortex before submitting them to the supply of C0 2 .
La invención también se refiere a la utilización 30 de C02 para, ventajosamente, aumentar la concentración en peso de metabolitos secundarios en las hojas de una especie vegetal.The invention also relates to the use 30 of C0 2 to advantageously increase the concentration by weight of secondary metabolites in the leaves of a plant species.
Más concretamente, la utilización de C02 da lugar 35 a un aumento de la concentración en peso de metabolitos
secundarios en las hojas igual o superior, en más de un 20%, a la concentración de metabolitos secundarios en las hojas antes de someterlas al suministro de C02.More specifically, the use of C0 2 results in an increase in the metabolite concentration by weight Secondary in the leaves equal or higher, by more than 20%, to the concentration of secondary metabolites in the leaves before submitting them to the supply of C0 2 .
Preferiblemente, la utilización de C02 da lugar a un aumento de la concentración en peso de metabolitos secundarios en las hojas igual o superior, en más de un 25%, a la concentración de metabolitos secundarios en las hojas antes de someterlas al suministro de C02.Preferably, the use of C0 2 results in an increase in the concentration by weight of secondary metabolites in the leaves equal or greater, by more than 25%, to the concentration of secondary metabolites in the leaves before subjecting them to the supply of C0 2 .
La presente invención también se refiere a la utilización de C02 en una dosis superior a 350 ppm, preferiblemente una dosis entre 500 y 700 ppm, para aumentar la concentración de metabolitos secundarios en una especie vegetal.The present invention also relates to the use of C0 2 in a dose larger than 350 ppm, preferably a dose between 500 and 700 ppm, to increase the concentration of secondary metabolites in a plant species.
Es evidente para un experto en la materia que el objeto de la presente invención puede llevarse a cabo con varias modificaciones sin salir del ámbito de la invención definida por las reivindicaciones.It is obvious to one skilled in the art that the object of the present invention can be carried out with several modifications without departing from the scope of the invention defined by the claims.
EJEMPLOSEXAMPLES
Ejemplo 1: Obtención de una especie vegetal modificada del género Taxus .Example 1: Obtaining a modified plant species of the genus Taxus.
A continuación se describe, a modo de ejemplo no limitativo, un procedimiento experimental para aumentar los niveles de taxanos en corteza y hojas.The following describes, by way of non-limiting example, an experimental procedure for increasing the levels of taxanes in bark and leaves.
En enero de 1997 se plantaron plántulas procedentes de una semilla comercial en un tiesto de 16 c de diámetro, en un sustrato compostado de corteza de pino.In January 1997, seedlings from a commercial seed were planted in a pot of 16 c in diameter, in a substrate composed of pine bark.
Los tejos son irrigados por aspersión dos veces al
dia con una solución nutritiva que comprende los nutrientes :P2θ5:K20, en una relación 1:0,5:1,5 y pH 6,5.Yews are irrigated by spraying twice at day with a nutrient solution comprising the nutrients: P 2 θ 5 : K 2 0, in a ratio 1: 0.5: 1.5 and pH 6.5.
Durante el periodo invernal (desde enero hasta mayo) se les suministró C02 en varios niveles (350, 500 y 700 ppm) , y a cada nivel de C02 se le suministraron diferentes regímenes hidricos (50 y 100%, 400 y 800 m-año"1 respectivamente). De esta forma los tejos se trataron en seis condiciones diferentes .During the winter period (from January to May) C0 2 was supplied at various levels (350, 500 and 700 ppm), and at each level of C0 2 different water regimes (50 and 100%, 400 and 800 m-) were supplied. year "1 respectively). Thus the yews were treated in six different conditions.
A partir de mayo, debido a la necesidad de ventilar el invernadero a causa de las altas temperaturas, se detuvo el suministro de C02. Y el régimen hidrico se aplicó mediante riego localizado con la misma solución, con unas dosis de agua de 116 1/m2 (tratamiento 100%) y 58 1/m2 (tratamiento 50%) .As of May, due to the need to ventilate the greenhouse due to high temperatures, the supply of C0 2 was halted. And the hydric regime was applied by irrigation located with the same solution, with water doses of 116 1 / m 2 (100% treatment) and 58 1 / m 2 (50% treatment).
La aplicación de C02 se realizó mediante tuberías de polietileno convencional. La fuente de C02 era un depósito de C02 líquido dotado de un evaporador para pasar el C02 a fase gas. La dosificación del C02 se realizó mediante un sistema compuesto por un analizador de infrarrojos, un autómata y una serie de electroválvulas que inyectaban C02 en función de las ordenes del autómata.The application of C0 2 was carried out using conventional polyethylene pipes. The source of C0 2 was a liquid C0 2 tank equipped with an evaporator to pass the C0 2 to the gas phase. The dosage of C0 2 was carried out by means of a system composed of an infrared analyzer, an automaton and a series of solenoid valves that injected C0 2 depending on the orders of the automaton.
Cada mes de mayo, después de finalizar la adición de CO2, se realizaron las medidas de la biomasa y taxanos presentes en las hojas y corteza.Each month of May, after finishing the addition of CO2, the measurements of the biomass and taxanes present in the leaves and bark were made.
Ejemplo 2: Efectos del enriquecimiento con C02 en el tejo.Example 2: Effects of enrichment with C0 2 on yew.
El procedimiento seguido para analizar los niveles de taxanos comprende las siguientes etapas:The procedure followed to analyze the levels of taxanes comprises the following stages:
1- Congelar hojas y tallos.
2- Liofilización.1- Freeze leaves and stems. 2- Lyophilization.
3- Conservación en seco.3- Dry preservation.
4- Análisis de la cantidad de taxanos mediante un ensayo inmuno enzimático Elisa (Heinstein y Chang, 1994) .4- Analysis of the amount of taxanes by an Elisa immunochemical assay (Heinstein and Chang, 1994).
Los resultados obtenidos se muestran en las figuras 1-8, de las que se deduce lo siguiente:The results obtained are shown in Figures 1-8, from which the following follows:
- El crecimiento del tejo debido al enriquecimiento con C02 es más marcado el primer año, desapareciendo el segundo año (Callaway et . al. 1994) .- Yew growth due to enrichment with C0 2 is more marked the first year, disappearing the second year (Callaway et. Al. 1994).
El abono carbónico no promueve ' diferencias significativas en la tasa de crecimiento relativo (RGR) de toda la planta de tejo ni de sus diferentes fracciones .The carbonic fertilizer does not promote 'significant differences in the relative growth rate (RGR) of all yew plant or its fractions.
El árbol muestra una tendencia positiva al crecimiento en presencia de niveles mayores de C02 durante los dos años .The tree shows a positive tendency towards growth in the presence of higher levels of C0 2 during the two years.
De esta manera se comprueba que durante el primer año el incremento de peso seco de hojas, tallos y raíces de las plantas crecidas a 500 ppm de C02 es mayor respecto a las crecidas a 350 ppm (27,8; 7,2; 48,5%). Cuando se suministra C02 en una dosis de 700 ppm el incremento en peso es menor siendo, incluso, negativo (-1,3; 9,2; 37,2%), como se ilustra en las figuras 9-12.In this way it is found that during the first year the increase in dry weight of leaves, stems and roots of plants grown at 500 ppm of C0 2 is greater compared to those grown at 350 ppm (27.8; 7.2; 48 ,5%). When C0 2 is supplied in a 700 ppm dose, the increase in weight is smaller, even being negative (-1.3; 9.2; 37.2%), as illustrated in Figures 9-12.
El efecto del agua en el crecimiento del tejo no muestra diferencias significativas aunque se muestra la tendencia que los valores en peso seco aumentan cuanta más cantidad de agua sea aportada.The effect of water on the growth of yew does not show significant differences although the tendency is shown that the values in dry weight increase the more water is supplied.
En cuanto a la producción de taxanos, al contrario de lo que sucede con la biomasa, sí que se muestran
diferencias significativas en los diferentes tratamientos con C02 y diferentes regímenes hídricos, y presenta una tendencia a aumentar la cantidad de taxanos a medida que aumenta la concentración de C02; así lo ilustran las figuras 13 y 14.As for the production of taxanes, contrary to what happens with biomass, they are shown significant differences in the different treatments with C0 2 and different water regimes, and it has a tendency to increase the amount of taxanes as the concentration of C0 2 increases ; this is illustrated by figures 13 and 14.
De esta manera, las plantas crecidas a 700 ppm contienen un 28,8% más de taxanos en 400 mm-año""1 y 49,36% más de taxanos en 800 mm-año"1 en las hojas respecto a las sometidas a 350 ppm.In this way, plants grown at 700 ppm contain 28.8% more taxanes in 400 mm-years "" 1 and 49.36% more taxanes in 800 mm-years "1 in the leaves compared to those submitted to 350 ppm
En la corteza, las plantas a las que se les suministró una dosis de C02 de 700 ppm presentaron un 152% más de taxanos en 400 mm-año"1 y un 517% más de taxanos en 800 mm-año-1 en la corteza respecto a las sometidas a 350 ppm.In the bark, plants that were given a dose of C0 2 of 700 ppm had 152% more taxanes in 400 mm-year "1 and 517% more taxanes in 800 mm-year -1 in the crust compared to those subjected to 350 ppm.
FIGURASFIGURES
Figuras 1 y 2: Tasa de crecimiento relativo (RGR) del peso seco de hojas a 400 y 800 mm-año"1, respectivamente .Figures 1 and 2: Relative growth rate (RGR) of dry leaf weight at 400 and 800 mm-year "1 , respectively.
Figuras 3 y 4: Tasa de crecimiento relativo (RGR) del peso seco de tallos a 400 y 800 mm-año"1, respectivamente.Figures 3 and 4: Relative growth rate (RGR) of the dry weight of stems at 400 and 800 mm-year "1 , respectively.
Figuras 5 y 6: Tasa de crecimiento relativo (RGR) del peso seco de raíces a 400 y 800 mm-año"1, respectivamente . Figuras 7 y 8: Tasa de crecimiento relativo (RGR) del peso seco total a 400 y 800 mm-año"1 , respectivamente.Figures 5 and 6: Relative growth rate (RGR) of the dry root weight at 400 and 800 mm-year "1 , respectively. Figures 7 and 8: Relative growth rate (RGR) of the total dry weight at 400 and 800 mm -year "1 , respectively.
Figura 9: Distribución del peso seco de hojas crecidas a tres dosis de C02 y dos de agua. Los valores son la media de cinco plantas ± error estándar. Donde : Línea continua: y= 0,007x+l,165 R2= 0,8334
Línea discontinua: y= 0, 0032x+4 , 0342 R2= 0,8358 Figura 10: Distribución del peso seco de raíces crecidas a tres dosis de C02 y dos de agua. Los valores son la media de cinco plantas ± error estándar. Donde :Figure 9: Distribution of dry weight of leaves grown at three doses of C0 2 and two of water. The values are the average of five plants ± standard error. Where: Continuous line: y = 0.007x + l, 165 R 2 = 0.8334 Dashed line: y = 0.0032x + 4, 0342 R 2 = 0.8358 Figure 10: Distribution of dry weight of roots grown at three doses of C0 2 and two of water. The values are the average of five plants ± standard error. Where :
Línea continua: y= 0, 0042x+5, 2618 R2= 0,6816 Línea discontinua: y= 0, 0154x+0, 7915 R2= 0,9519 Figura 11: Área foliar de las especies vegetales de Taxus crecidas a tres dosis de C02 y dos de agua. Los valores son la media de cinco plantas ± error estándar. Donde :Continuous line: y = 0.0042x + 5, 2618 R 2 = 0.6816 Dashed line: y = 0.0154x + 0.7915 R 2 = 0.9519 Figure 11: Leaf area of plant species of Taxus grown to three dose of C0 2 and two of water. The values are the average of five plants ± standard error. Where :
Línea continua: y= -0, 0507x+550, 9 R2= 0,6401 Línea discontinua: y= 0, 0154x+0, 7915 R2= 0,9933 Figura 12: Relación raíz/copa de las plantas de Taxus , crecidas a tres dosis de C02 y dos de agua. Los valores son la media de 5 plantas ± error estándar. Donde :Continuous line: y = -0.0507x + 550, 9 R 2 = 0.6401 Dashed line: y = 0.0154x + 0.7915 R 2 = 0.9933 Figure 12: Root / cup ratio of Taxus plants, grown at three doses of C0 2 and two of water. The values are the average of 5 plants ± standard error. Where :
Línea continua: y= -0, 0001x+l, 1577 R2= 0,5793 Línea discontinua: y= 0,0009x4-0,6029 R2= 0,4718 Figura 13: Concentración de taxanos en las hojas de las especies vegetales del género Taxus crecidas en tres concentraciones de C02 y dos dosis de agua. Los valores son la media de 2 valores ± error estándar. Donde : Línea continua: y= 0, 0019x+l, 0841 R2= 0,3335 Línea discontinua: y= 0, 0023x+0, 6103 R2= 0,7737 Figura 14: Concentración de taxanos en la corteza de los tallos de las especies vegetales del género Taxus crecidas en tres concentraciones de C02 y dos dosis de agua. Los valores son la media de 2 valores + error estándar . Donde :Continuous line: y = -0, 0001x + l, 1577 R 2 = 0.5793 Dashed line: y = 0.0009x4-0.6029 R 2 = 0.4718 Figure 13: Concentration of taxanes in the leaves of plant species of the genus Taxus grown in three concentrations of C0 2 and two doses of water. The values are the average of 2 values ± standard error. Where: Continuous line: y = 0.0019x + l, 0841 R 2 = 0.3335 Dashed line: y = 0.0023x + 0, 6103 R 2 = 0.7737 Figure 14: Concentration of taxanes in the bark of the stems of the plant species of the genus Taxus grown in three concentrations of C0 2 and two doses of water. The values are the average of 2 values + standard error. Where :
Línea continua: y= 0, 0027x-0, 6655 R2= 0,531 Línea discontinua: y= 0, 00834x-2, 4862 R2= 0,9817
Continuous line: y = 0.0027x-0, 6655 R 2 = 0.531 Dashed line: y = 0.00834x-2, 4862 R 2 = 0.9817
Claims
R E I V I N D I C A C I O N E S R E I V I N D I C A C I O N E S
- Especie vegetal modificada del género Taxus caracterizada por el hecho que comprende una concentración en peso de taxanos superior a la concentración en peso de taxanos en una especie vegetal no modificada del género Taxus .- A modified plant species of the genus Taxus characterized by the fact that it comprises a concentration by weight of taxanes greater than the concentration by weight of taxanes in an unmodified plant species of the genus Taxus.
- Especie vegetal según la reivindicación 1, caracterizada por el hecho que la corteza comprende una concentración en peso de taxanos superior a la concentración en peso de taxanos en la corteza de una especie vegetal no modificada del género Taxus .- Vegetable species according to claim 1, characterized in that the bark comprises a concentration by weight of taxanes in excess of the concentration by weight of taxanes in the bark of an unmodified plant species of the genus Taxus.
- Especie vegetal según la reivindicación 1 o 2, caracterizada por el hecho que la corteza comprende una concentración en peso de taxanos igual o superior, en más de un 50%, a la concentración en peso de taxanos en la corteza de una especie vegetal no modificada del género Taxus .- Plant species according to claim 1 or 2, characterized in that the bark comprises a concentration by weight of taxanes equal to or greater than 50%, at the concentration by weight of taxanes in the bark of a non-plant species modified from the genus Taxus.
- Especie vegetal según la reivindicación 3, caracterizada por el hecho que en la corteza la concentración en peso de taxanos es, preferiblemente, igual o superior, en más de un 100%, a la concentración en peso de taxanos en la corteza de una especie vegetal no modificada del género Taxus .- Vegetable species according to claim 3, characterized in that in the bark the concentration by weight of taxanes is, preferably, equal or greater, by more than 100%, to the concentration by weight of taxanes in the bark of a species unmodified vegetable of the genus Taxus.
- Especie vegetal según la reivindicación 1, caracterizada por el hecho que en las hojas la concentración en peso de taxanos es superior a la concentración en peso de taxanos en las hojas de una especie vegetal no modificada del género Taxus .- Vegetable species according to claim 1, characterized in that in the leaves the concentration by weight of taxanes is higher than the concentration by weight of taxanes in the leaves of an unmodified plant species of the genus Taxus.
- Especie vegetal según la reivindicación 1 o 5,
caracterizada por el hecho que en las hojas la concentración en peso de taxanos es igual o superior, en más de un 20%, a la concentración en peso de taxanos en las hojas de una especie vegetal no modificada del género Taxus.- Vegetable species according to claim 1 or 5, characterized by the fact that in the leaves the concentration by weight of taxanes is equal or greater, by more than 20%, to the concentration by weight of taxanes in the leaves of an unmodified plant species of the genus Taxus.
- Especie vegetal según la reivindicación 6 caracterizada por el hecho que en las hojas la concentración en peso de taxanos es, preferiblemente, igual o superior, en más de un 25%, a la concentración en peso de taxanos en las hojas de una especie vegetal no modificada del género Taxus- Vegetable species according to claim 6 characterized in that in the leaves the concentration by weight of taxanes is, preferably, equal to or greater than 25%, at the concentration by weight of taxanes in the leaves of a plant species unmodified of the genus Taxus
- Procedimiento de obtención de una especie vegetal modificada del género Taxus que se caracteriza por someter dicha especie vegetal del género Taxus a un régimen hídrico de entre el 50 y el 100% y a una dosis de C02 superior a 350 ppm.- Procedure for obtaining a modified plant species of the genus Taxus that is characterized by subjecting said plant species of the genus Taxus to a water regime of between 50 and 100% and at a dose of C0 2 greater than 350 ppm.
- Procedimiento según la reivindicación 8, caracterizado por el hecho que el régimen hídrico. preferiblemente, es del 100% y la dosis de C02 oscila, preferiblemente, entre 500-700 ppm.- Procedure according to claim 8, characterized by the fact that the water regime. preferably, it is 100% and the dose of C0 2 preferably ranges between 500-700 ppm.
0- Utilización de C02 para incrementar la concentración en peso de metabolitos secundarios en una especie vegetal.0- Use of C0 2 to increase the concentration by weight of secondary metabolites in a plant species.
1- Utilización según la reivindicación 10, caracterizada por el hecho que el incremento de la concentración en peso de metabolitos secundarios tiene lugar en las hojas, corteza, tallos, ramas, flores o raíces de la especie vegetal1- Use according to claim 10, characterized in that the increase in the concentration by weight of secondary metabolites takes place in the leaves, bark, stems, branches, flowers or roots of the plant species
2- Utilización según la reivindicación 11,
caracterizada por el hecho que el incremento de la concentración en peso de metabolitos secundarios en la corteza es igual o superior, en más de un 50%, a la concentración en peso de metabolitos secundarios en la corteza antes de someterlas al suministro de C02.2- Use according to claim 11, characterized by the fact that the increase in the concentration by weight of secondary metabolites in the cortex is equal or greater, by more than 50%, to the concentration by weight of secondary metabolites in the cortex before subjecting them to the supply of C0 2 .
- Utilización según la reivindicación 12, caracterizada por el hecho que el aumento de la concentración en peso de metabolitos secundarios en la corteza es igual o superior, en más de un 100%, a la concentración en peso de metabolitos secundarios en la corteza antes de someterlas al suministro de C02.- Use according to claim 12, characterized in that the increase in the concentration by weight of secondary metabolites in the cortex is equal to or greater than, by more than 100%, the concentration by weight of secondary metabolites in the cortex before submit them to the supply of C0 2 .
- Utilización según la reivindicación 11, caracterizada por el hecho que el incremento de la concentración en peso de metabolitos secundarios en las hojas es igual o superior, en más de un 20%, a la concentración en peso de metabolitos secundarios en las hojas antes de someterlas al suministro de C02.- Use according to claim 11, characterized in that the increase in the weight concentration of secondary metabolites in the leaves is equal to or greater than, by more than 20%, the concentration in weight of secondary metabolites in the leaves before submit them to the supply of C0 2 .
- Utilización según la reivindicación 14, caracterizada por el hecho que el incremento de la concentración en peso de metabolitos secundarios en las hojas es, preferiblemente, igual o superior, en más de un 25%, a la concentración en peso de metabolitos secundarios en las hojas antes de someterlas al suministro de C02.- Use according to claim 14, characterized in that the increase in the concentration by weight of secondary metabolites in the leaves is, preferably, equal to or greater than, by more than 25%, to the concentration by weight of secondary metabolites in the leaves. sheets before submitting them to the supply of C0 2 .
- Utilización según cualquiera de las reivindicaciones 10 a 15, caracterizada por el hecho que la dosis de C02 utilizada es superior a 350 ppm.- Use according to any of claims 10 to 15, characterized in that the dose of C0 2 used is greater than 350 ppm.
- Utilización según la reivindicación 16, caracterizada por el hecho que la dosis de C02 oscila, preferiblemente, entre 500-700 ppm.
- Use according to claim 16, characterized in that the dose of C0 2 preferably ranges between 500-700 ppm.
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| ESP200101951 | 2001-08-23 | ||
| ES200101951A ES2183745B1 (en) | 2001-08-23 | 2001-08-23 | TAXUS PLANT M0DIFIED AND PROCEDURE FOR OBTAINING. |
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| WO2003017754A1 true WO2003017754A1 (en) | 2003-03-06 |
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| WO (1) | WO2003017754A1 (en) |
Cited By (5)
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|---|---|---|---|---|
| CN103430740A (en) * | 2013-08-14 | 2013-12-11 | 江苏鼎钰生态农业科技有限公司 | Method for planting taxus chinensis in saline and alkaline land |
| CN104838948A (en) * | 2015-05-14 | 2015-08-19 | 平顶山学院 | Planting method of pure ecological taxus chinensis |
| CN106550837A (en) * | 2015-09-29 | 2017-04-05 | 奉节县奎满农业开发有限公司 | The nursery fertilizing method of Chinese yew |
| CN106550838A (en) * | 2015-09-29 | 2017-04-05 | 奉节县奎满农业开发有限公司 | The implantation methods of Ramulus et folium taxi cuspidatae |
| CN111492885A (en) * | 2020-05-14 | 2020-08-07 | 东北林业大学 | How to build a mixed forest of yew and fig trees |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103430740A (en) * | 2013-08-14 | 2013-12-11 | 江苏鼎钰生态农业科技有限公司 | Method for planting taxus chinensis in saline and alkaline land |
| CN104838948A (en) * | 2015-05-14 | 2015-08-19 | 平顶山学院 | Planting method of pure ecological taxus chinensis |
| CN106550837A (en) * | 2015-09-29 | 2017-04-05 | 奉节县奎满农业开发有限公司 | The nursery fertilizing method of Chinese yew |
| CN106550838A (en) * | 2015-09-29 | 2017-04-05 | 奉节县奎满农业开发有限公司 | The implantation methods of Ramulus et folium taxi cuspidatae |
| CN111492885A (en) * | 2020-05-14 | 2020-08-07 | 东北林业大学 | How to build a mixed forest of yew and fig trees |
| WO2021227168A1 (en) * | 2020-05-14 | 2021-11-18 | 东北林业大学 | Method for constructing mixed forest of taxus chinensis trees and ficus carica trees |
| US11240955B2 (en) | 2020-05-14 | 2022-02-08 | Northeast Forestry University | Method for constructing a mixed forest of yew trees and fig trees |
Also Published As
| Publication number | Publication date |
|---|---|
| ES2201933B1 (en) | 2005-02-01 |
| ES2183745B1 (en) | 2004-06-01 |
| ES2183745A1 (en) | 2003-03-16 |
| ES2201933A1 (en) | 2004-03-16 |
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