U.S. patent application number 10/178768 was filed with the patent office on 2003-06-05 for alkaline preparaions of inula for the control of fungal diseases in plants.
Invention is credited to Cohen, Yigal.
Application Number | 20030104086 10/178768 |
Document ID | / |
Family ID | 26753281 |
Filed Date | 2003-06-05 |
United States Patent
Application |
20030104086 |
Kind Code |
A1 |
Cohen, Yigal |
June 5, 2003 |
Alkaline preparaions of Inula for the control of fungal diseases in
plants
Abstract
Rapid, convenient and inexpensive methods for preparing
fungicidal suspensions from Inula are described. Methods for
controlling fungal disease and crops by using alkaline solutions
are also disclosed.
Inventors: |
Cohen, Yigal; (Kiryat Ono,
IL) |
Correspondence
Address: |
SOL SHEINBEIN
c/o ANTHONY CASTORINA
SUITE 207
2001 JEFFERSON DAVIS HIGHWAY
ARLINGTON
VA
22202
US
|
Family ID: |
26753281 |
Appl. No.: |
10/178768 |
Filed: |
June 25, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10178768 |
Jun 25, 2002 |
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09781164 |
Feb 13, 2001 |
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6423352 |
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09781164 |
Feb 13, 2001 |
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09233971 |
Jan 20, 1999 |
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60072356 |
Jan 23, 1998 |
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Current U.S.
Class: |
424/764 |
Current CPC
Class: |
A01N 65/12 20130101;
A01N 25/02 20130101; A01N 65/12 20130101 |
Class at
Publication: |
424/764 |
International
Class: |
A61K 035/78 |
Claims
What is claimed:
1. A method for preparing an extract from Inula species, said
method comprising the steps of: (a) contacting shoots and/or
portions thereof with an alkaline aqueous solvent to form a
solution and debris; and (b) removing the debris from the
solution.
2. A method according to claim 1, wherein said removing step
includes filtering the debris from the solution.
3. A method according to claim 1, wherein the shoots include leaves
and stems of upper parts of the shoots.
4. A method according to claim 1, wherein the shoots or portions
thereof are dried prior to said contacting step.
5. A method according to claim 4 including the step of grinding the
dried shoots or portions thereof to form a powder.
6. A method according to claim 1, wherein the shoots or portions
thereof are freshly cut.
7. A method according to claim 1, wherein the alkaline aqueous
solvent comprises at least one amino compound.
8. A method according to claim 7, wherein the amino compound
comprises an ammonium compound.
9. A method according to claim 7, wherein the amino compound is
selected from the group consisting essentially of organic amines
and heterocyclic amines.
10. A method according to claim 9, wherein the organic amine
compound is selected from the group consisting essentially of
ethylamine, ethylenediamine, diaminopropane, diethanolamine,
triethanolamine, and butylamine.
11. A method according to claim 9, wherein the heterocyclic amine
compound is selected from the group consisting essentially of
piperazine and piperidine.
12. A method according to claim 7, wherein the alkaline aqueous
solvent comprises inorganic bases or salts thereof.
13. A method for protecting plants against fungal infection, said
method comprising the steps of: (a) preparing a fungicidal extract
of Inula species by contacting shoots and/or portions thereof with
an alkaline aqueous solvent to form an extract solution and debris;
and (b) applying a fungicidally effective amount of the fungicidal
extract to a plant for protecting the plant against fungal
infection.
14. A method according to claim 13, wherein the plant includes
grapevines, cucurbits, tomatoes, wheat, barley, onion, tobacco,
crucifers, and potatoes.
15. A method according to claim 13, wherein the fungal infection is
caused by phytopathogenic fungi including Oomycetes, Ascomycetes,
Basidiomycetes and Fungi imperfecti classes.
16. A method according to claim 15, wherein the fungal infection is
caused by a fungus including Cladosporium cucumerinum, Phytophthora
infestans, Botrytis cinerea, Pseudoperonospora cubensis,
Sphaerotheca fuliginea, Plasmopara viticola, Uromyces
appendiculatis, and Erysiphe graminis.
17. A method according to claim 13, wherein said preparing step
includes filtering the debris from the solution.
18. A method according to claim 13, wherein the shoots include
leaves and stems of upper parts of the shoots.
19. A method according to claim 13, wherein the shoots or portions
thereof are dried prior to said contacting step.
20. A method according to claim 19 including the step of grinding
the dried shoots or portions thereof to form a powder.
21. A method according to claim 13, wherein the shoots or portions
thereof are freshly cut.
22. A method according to claim 13, wherein the alkaline aqueous
solvent comprises at least one amino compound.
23. A method according to claim 22, wherein the amino compound
comprises an ammonium compound.
24. A method according to claim 22, wherein the amino compound is
selected from the group consisting essentially of organic amines
and heterocyclic amines.
25. A method according to claim 24, wherein the organic amine
compound is selected from the group consisting essentially of
ethylamine, ethylenediamine, diaminopropane, diethanolamine,
triethanolamine, and butylamine.
26. A method according to claim 24, wherein the heterocyclic amine
compound is selected from the group consisting essentially of
piperazine and piperidine.
27. A method according to claim 13, wherein the alkaline aqueous
solvent comprises inorganic bases or salts thereof.
28. A method for preparing a fungicide derived from Inula species,
said method comprising the step of: combining a substantially
powdered form of Inula shoots and/or portions thereof with a solid
chemical to form a mixture which upon dissolution of the mixture in
an aqueous solution, forms an alkaline aqueous solution which can
be directly applied to plants to prevent fungal infection.
29. A method according to claim 28, wherein the solid chemical
comprises at least one amino compound.
30. A method according to claim 29, wherein the amino compound
comprises an ammonium compound.
31. A method according to claim 29, wherein the amino compound is
selected from the group consisting essentially of organic amines
and heterocyclic amines.
32. A method according to claim 31, wherein the organic amine
compound is selected from the group consisting essentially of
ethylamine, ethylenediamine, diaminopropane, diethanolamine,
triethanolamine, and butylamine.
33. A method according to claim 31, wherein the heterocyclic
compound is selected from the group consisting essentially of
piperazine and piperdine.
34. A method according to claim 28, wherein the solid chemical
comprises inorganic bases or salts thereof.
35. A method according to claim 28, wherein an emulsifier is added
to the mixture.
36. A method according to claim 35, wherein the emulsifier
comprises a surfactant.
37. A method according to claim 36, wherein the surfactant
comprises sodium dodecyl sulfate.
38. A method according to claim 35, wherein the emulsifier
comprises diatomaceous earth.
Description
[0001] This application claims priority of U.S. Provisional
Application Serial No. 60/072,356 filed Jan. 23, 1998.
FIELD OF THE INVENTION
[0002] The subject invention relates an anti-fungal preparation for
the control of fungal diseases in plants, more specifically, to
alkaline extracts of Inula which are highly active in controlling
diseases caused by fungi in crop plants.
BACKGROUND OF THE INVENTION
[0003] Extracts of plants which are members of the Inula species
are effective against infections of plants caused by a variety of
fungi. These extracts are typically prepared by dipping freshly cut
Inula shoots in an organic solvent or by agitating freshly cut or
dried Inula shoots in an organic solvent, removing the solvent to
form a paste, and then dissolving the paste in an organic solvent
or in water, possibly with an additive. Alternatively, suspensions
of Inula can be prepared by grinding dried Inula shoots into a fine
powder, adding an emulsifier, and then suspending the mixture in
water. In both cases, the resultant preparation is then applied to
plants resulting in the control of a wide variety of fungal
diseases. Extracts of suspensions of Inula plants are effective at
low concentrations, in the range of fractions of a single percent
of extract, such that dilute concentrations have excellent
fungal-control properties.
[0004] The plants which are the basis of these fungicidal extracts
and suspensions are Inula viscosa and Inula graveolens (Family
Compositae), perennial weeds widespread in the Mediterranean
Basin.
[0005] Methods of preparing aqueous extracts from various parts of
the Inula plant are well known in the literature. Additionally,
organic extracts of the Inula plant are also well known in the art,
e.g., U.S. Pat. No. 5,837,253. However, the prior art indicates
that aqueous extracts of Inula shoots are poorly effective against
fungal diseases of crop plants whereas extracts made with organic
solvents are highly effective anti-fungal agents.
[0006] Methods have also been described for using organic solvents
to extract Inula plants, but these are clearly distinguished from
that disclosed in U.S. Pat. No. 5,837,253. Two of the prior art
methods involved contacting the whole Inula plant, or the aerial
parts thereof, with an organic solvent either by maceration of the
plant in the organic solvent or by percolation of the solvent
through the plant. Furthermore, U.S. Pat. No. 4,254,112 to Debat et
al., (hereinafter referred to as "Debat") describes the preparation
of extracts of Inula viscosa and Inula graveolens using whole Inula
plants which have been dried and ground and organic solvents, by
using a Soxhlet apparatus. The yield of the paste obtained by this
method was approximately 1.75-4%. U.S. Pat. No. 5,176,193 to
Honerlagen et al. (hereinafter referred to as "Honerlagen")
describes a process for preparing a partial extract from roots of
Inula helenium which involves contacting the plant material with an
organic solvent, adding a drying agent to the solution to remove
the water, removing the drying agent, and then distilling the dried
organic phase. By contrast, the method disclosed in U.S. Pat. No.
5,837,753 disclosed either briefly dipping the leaves and stems of
the shoots of Inula viscosa or Inula graveolens into an organic
solvent or shaking the freshly cut or dried and ground leaves and
stems of the shoots in an organic solvent for thirty minutes, and
then evaporating the solvent to form a paste. The yield obtained by
this method can be as much as 30%, in contrast to the low yields
known in the literature and described above.
[0007] The medicinal properties of Inula extracts in humans are
well known. For example, Debat disclosed the anti-microbial
activity of extracts of Inula for use in human beings. However, the
fungicidal effects of Inula extracts have only been demonstrated on
fungi growing in Petri dishes or on post-harvest of fruits. For
example, Qasem et al. (Phytopathologia Mediterana, 34:7-14, 1995)
demonstrated that the growth of certain fungi in Petri dishes was
inhibited by aqueous extracts of Inula viscosa as well as by dried
plant material added directly to the fungal growth media. By
contrast, the method of the present invention uses Inula extracts
prepared with alkaline aqueous solvents used against fungal
infections of crop plants themselves.
[0008] Clearly, although Inula extracts have been shown to have
fungicidal activity in the Petri dish (in vitro) and on plant (in
vivo), the methods of preparation for these extracts have not been
sufficient for large scale use directly on crop plants and have had
other serious disadvantages. The true effectiveness of these
extracts against fungal infections of plants is, therefore,
unknown. Furthermore, there is a clear need for better methods to
prepare Inula extracts. Qasem et al. (Ibid, page 13, 1995)
concluded: "The diversity and the methodology of extraction and the
differences in the results obtained . . . increased the need for
developing more efficient, convenient, and cheaper methods of
extraction to facilitate more extensive utilization of fungicidal
extracts, especially if greater quantity of extracts must be
prepared for large-scale production".
[0009] Thus, there is a widely recognized need for and would be
highly advantageous to have a method or methods for preparing
extracts and/or suspensions of plants of Inula species which would
facilitate the large-scale use of these extracts and suspensions
which would simplify their preparation and use, be highly effective
in controlling fungal infection in plants, as well as controlling
fungal infections in crop plants.
SUMMARY OF THE INVENTION
[0010] According to the present invention, there is provided a
method for preparing an aqueous extract from Inula species which
comprises contacting shoots and/or portions thereof with an
alkaline aqueous solvent to form a solution and debris, and
removing the debris from the solution.
[0011] There is also provided a method for protecting plants
against fungal infection comprising preparing a fungicidal extract
of Inula species by contacting shoots and/or portions thereof with
an alkaline aqueous solvent to form an extract solution and debris,
and applying a fungicidally effective amount of the fungicidal
extract to a plant for protecting the plant against fungal
infection.
[0012] There is also provided in accordance with the present
invention, a method for preparing a fungicide derived from Inula
species which comprises combining a substantially powdered form of
Inula shoots and/or portions thereof with a solid chemical to form
a mixture which upon dissolution of the mixture in an aqueous
solvent, forms an alkaline aqueous solution which can be directly
applied to plants to prevent fungal infection.
[0013] Additionally, also in accordance with the present invention,
there is provided an alkaline anti-fungal composition which
comprises Inula shoots and/or portions thereof and a compound which
when mixed with an aqueous solvent forms an alkaline solution,
whereby the composition can be directly applied to plants to
prevent fungal infection.
[0014] Accordingly, the subject invention provides a method for
unexpectedly producing an anti-microbial extract from Inula plants
which is not only active in vitro against fungi but also is active
against diseases caused by fungi in crop plants while being safe to
apply to the crop itself.
DETAILED DESCRIPTION OF THE INVENTION
[0015] The present invention provides a method for preparing an
extract from Inula species including the steps of contacting shoots
and/or portions thereof with an alkaline aqueous solvent to form a
solution and debris and then removing the debris from the
solution.
[0016] Preferably, the portion and/or portions of the Inula plant
which are utilized in the present invention are the leaves and stem
of the upper, younger parts of the shoots, and the upper part of
the shoot extending from about 20-40 centimeters from the tip of
the shoot. The flowers themselves are not used. As used herein, the
term "shoot" is used to define the leaves and stem of the shoot of
the Inula plant. In the methods of the present invention, the
shoots or portions thereof can be either freshly cut or may be
dried prior to the addition of the alkaline aqueous solvent.
[0017] The alkaline aqueous solvent can include any compound or
compounds which alone or in solution have a pH which is greater
than about seven such as an inorganic base or salt thereof.
[0018] Preferably, the alkaline aqueous solvent includes at least
one of amino compound. The amino compound can include an ammonium
compound, organic amines and/or heterocyclic amines. The organic
amines can include compounds such as ethylamine, ethylenediamine,
diaminopropane, diethanolamine, triethanolamine, and butylamine.
Heterocyclic compounds can include compounds such as piperazine and
piperidine.
[0019] The removal of the debris from the solution can be
accomplished by filtering the debris from the solution utilizing a
screen such as a 100 micron .mu.m (micron) sieve or other suitable
filtering devices known to those skilled in the art.
[0020] If dried shoots and/or portions thereof are utilized in the
method of the present invention, the dried shoots and/or portions
thereof can be ground, by various means known to those skilled in
the art, to form a powder.
[0021] In an embodiment of the invention, fresh Inula shoots are
dipped in the alkaline solution for an appropriate period of time,
typically for approximately 0.2-5 minutes. The plant material is
then discarded and the solution, which may be colored, obtained
therefrom is screened through a filter such as a 100 micron sieve
and can be applied, as is, for spraying crop plants.
[0022] In another embodiment, fresh shoots are first air dried at
30-40.degree. C. for period of time sufficient to dry the shoots,
typically approximately 1-2 days and the dried shoots then dipped
in the alkaline solution for a period of time which can be
approximately 1-10 minutes. The plant material is discarded and the
solution (colored) obtained can be filtered through a filter or
sieve and then utilized, as is, for spraying crop plants.
[0023] In another embodiment, dried shoots obtained by air drying
as described immediately above can be ground and then passed
through a metal sieve (i.e., 2 mm). The ground plant material can
then be shaken in an alkaline solution for 1-10 minutes, screened
through a filtering device such as a 100 micron sieve, and the
colored liquid obtained therefrom can be utilized for spraying crop
plants.
[0024] In yet another embodiment, dried and ground plant material
as immediately described above, can be placed in a mortar and is
milled to less than 100 micron-sized particles to obtain a fine
powder. The powder is then mixed with an alkaline solution and can
be sprayed, as such, onto the crop plants.
[0025] In another embodiment, dried powder which is ground to less
than 100 micron-sized particles is milled with crystallized
cellulose (Avicel PH-101, Fluka) and is then mixed with the
alkaline solution and sprayed directly onto crop plants.
[0026] In another embodiment, a dried powder which has been milled
to less than 100 micron-sized particles is milled with a surfactant
such as Vercoryl-S, diatomaceous earth, or sodium dodecyl sulfate
and is then mixed with the alkaline solution and is sprayed
directly onto the crop plants.
[0027] In another embodiment, dried Inula shoots ground to less
then 100 micron-sized particles are mixed with a solid chemical,
which upon dissolution in water, produces an alkaline solution. The
solid chemical can be any of the compounds described above for the
alkaline solution including an imidazole, dibasic ammonium
phosphate, Trizma-base (Sigma) either with or without the addition
of an inert surfactant formulation. The powder mixture is then
mixed with water to form an anti-fungal agent and is sprayed
directly onto crop plants. In this form, the anti-fungal agent can
be prepared and/or sold as a dry mix which, when hydrated, forms an
anti-fungal composition which can be easily applied to plants. In
this form, the costs of shipping and/or transporting the
anti-fungal material can be reduced since the water would only be
added just prior to use/application to plants.
[0028] In another embodiment, the dried and ground Inula of
particle size less than 100 microns can be mixed with
(NH4).sub.2HPO.sub.4 (dibasic ammonium phosphate) and be suspended
in water to which NH.sub.4OH is added. This solution can then be
sprayed directly onto crop plants.
[0029] According to the present invention, there is also provided a
method for protecting plants against fungal infections by the steps
of (a) preparing a fungicidal extract of Inula species by
contacting shoots and/or portions thereof with an alkaline aqueous
solvent to form an extract solution and debris and (b) applying a
fungicidally effective amount of the fungicidal extract to a plant
for protecting a plant against fungal infection. As defined herein,
a fungicidally effective amount includes such an amount that is
capable of preventing, inhibiting, and/or killing fungus in or on a
plant.
[0030] Solutions or emulsions containing the Inuta extract, or the
Inula suspension, are sprayed onto the upper leaf surfaces of
various crop plants by well known techniques for the control of
fungal plant infections. The Inula extracts of the present
invention can be used to control fungal plant infections caused by
fungi including, but not limited to, Oomycetes, Ascomycetes,
Basidiomycetes, and Fungi imperfecti classes.
[0031] The method and fungicidal extracts of the present invention
are particularly suitable for use against fungi of the Oomycetes,
Ascomycetes, Basidiomycetes and Fungi imperfecti classes including
but not limited to Phytophthora infestans, Pseudoperonospora
cubensis, Plasmopara viticola, Sphaerotheca fuliginea, Cladosporium
cucumerinum, Erysiphe graminis, Uromyces appendiculatus, and
Botrytis cinera and in crops including, but not limited to,
grapevines, tomato, wheat, barley, tobacco, potatoes, onions,
cucurbits, beans and crucifers.
[0032] Also according to the present invention, there is provided a
method for preparing a fungicide derived from Inula species which
includes combining a substantially powdered form of Inula shoots
and/or portions thereof with a solid chemical to form a mixture
which upon dissolution of the mixture and an aqueous solution,
forms an alkaline aqueous solution which can be directly applied to
plants to prevent fungal infection. Fungicides prepared by this
method are described above and in the Example section below.
[0033] Also in accordance with the present invention, an alkaline
anti-fungal composition of Inula shoots and/or portions thereof and
a compound which when mixed with an aqueous solvent forms an
alkaline solution which can be directly applied to plants to
prevent fungal infection is disclosed. The compounds which can be
mixed with an aqueous solution to form an alkaline solution are
described above and in the Examples section below.
[0034] The invention of the present invention is illustrated below
in the following Examples which describe the preparation and use of
Inula extracts and suspensions against fungal infections in
plants.
[0035] To test the efficacy of these extracts and suspensions,
experiments were performed in which plants were sprayed using a
fine glass atomizer with either Inula extract or with pure solvent
as a control for the Inula extracts, with either Inula extract and
emulsifier or with an emulsifier solution as a control for the
emulsified Inula extracts, and with either Inula suspension or with
water or emulsifier solution as a control for the Inula
suspensions. Treated and control plants were then inoculated with a
crop-respective fungal pathogen. After an incubation period, the
extent of the infection was measured. Unless otherwise stated,
percentage protection from the disease due to the treatment with
the Inula extract or suspension was calculated as:
% control of the disease=[1-(% infection and treated plants/%
infection in control plants)].times.100.
[0036] The following experimental data illustrate the utility of
the flungicidal compositions of the present invention.
EXAMPLES
[0037]
1 METHODS: The crop plants and pathogens used for inoculation are
described in the following list. crop plant pathogen disease Tomato
Phytophthora infestans late blight Potato " " Cucumber
Pseudoperonospora cubensis downy mildew Melon " " Wheat Erysiphe
graminis tritici powdery mildew Cucumber Sphaerotheca fuliginea "
Melon " " Tobacco Perenospora tabacina downy mildew Grapes
Plasmpara viticola " Tomato Botrytis cinerea gray mold Cucumber " "
Tomato Cladosporium fulvum leaf mold Inoculation with P.
infestants, P. cubensis, P. tabacina, P. viticola, B. cinera and C.
fulvum was done by spraying a spore suspension in distilled water
onto the upper leaf surfaces of the treated (and the control)
plants. Inoculated plants were kept in a dew chamber at
18-20.degree. C. in the dark for 16-20 hours and then in a growth
chamber at 20.degree. C. with 12 hours photoperiod. Inoculation
with E. graminis and S. fuliginea was # done by blowing spores over
the treated (and control) plants. Inoculated plants were then kept
in a growth chamber as above. Disease records were visually
estimated in the inoculated plants at 3-13 days after inoculation,
depending on the crop and disease. Percentage leaf area occupied
with disease symptoms or fungal colonies was recorded and given in
the results section.
Example 1
[0038]
2 Activity of dipping extract of fresh Inula shoots in 0.1 N NaOH
for ten minutes on disease development in crop plants. % Diseased
leaf area powdery mildew Extract late blight in potato in wheat
control untreated 94 63 original (pH 12.7) 15 24 diluted: 2 9 25
diluted: 4 10 30 diluted: 8 8 50 original (neutralized to pH 7.2) 5
12 diluted: 2 94 18 diluted: 4 95 25 diluted: 8 95 30 50 g fresh
weight shoots were dipped in 250 ml of 0.1 N NaOH for ten minutes.
Half of the extract was sprayed (pH 12.7) after various dilutions
and the other half was neutralized with HCl to pH 7.2 and then
sprayed after various dilutions. Late blight and powdery mildew
were inoculated one day after spray and disease records taken after
four and eight days, respectively.
Example 2
[0039]
3 Activity of dipping extracts of fresh Inula shoots in ammonium
hydroxide (containing 25% NH.sub.3 by weight) for ten minutes on
late blight development in potato. Treatment Diseased leaf area %
Protection untreated control 98 -- NH.sub.4OH 8% 100 0 (=2% aqueous
NH.sub.3) 4% 100 0 2% 100 0 1% 100 0 Inula in NH.sub.4OH 8% 10 90
4% 20 80 2% 50 50 1% 88 12 60 g fresh weight shoot were dipped in
250 ml NH.sub.4OH solution for ten minutes. Extracts were sprayed
after various dilutions onto potato plants and inoculated with
Phytophtera infestants two days later. Disease was recorded four
days post inoculation. NH.sub.4OH solutions of corresponding
concentration were also sprayed for comparison. The % NH.sub.4OH
throughout the Examples, where appropriate, refers to # % from a
liquid containing 250 g NH.sub.3 per liter. For example, 8%
NH.sub.4OH means 8 ml of a solution (containing 25% NH.sub.3) per
100 ml water. To express % NH.sub.4OH as % NH.sub.3, should divide
by 4. Note: NH.sub.4OH was not phytotoxic.
Example 3
[0040]
4 Efficacy of Inula extracts, made by dipping fresh shoots in
NH.sub.4OH for ten minutes, in the control of late blight in potato
(a) and powdery mildew in wheat (b). % blighted leaf area 3d - old
extract NH.sub.4OH conc., % fresh extract 1d - old extract a b
untreated control 100 100 100 75 1 23 63 25 23 2 5 13 23 1 4 8 18 8
11 8 8 10 25 24 16 3 10 5 6 One Inula shoot (10 g fresh weight) was
dipped in 50 ml of NH.sub.4OH of various concentrations for ten
minutes and the extract sprayed onto potato plants one or three
days later. Plants were inoculated with P. infestants or E.
graminis one day later and disease records taken seven days post
inoculation.
Example 4
[0041]
5 Activity, after storage, of fresh shoot Inula extract made with
ammonium hydroxide against fungal diseases of crop plants. %
diseased leaf area downy powdery downy storage late blight mildew
in mildew mildew in NH.sub.4OH period, in potato, melon in wheat
grapes conc., % days 5 d 6 d 7 d 10 d control 1 98 81 100 50
untreated 0.25 38 15 50 20 0.5 15 4 13 15 1.0 8 1 1 5 control 9 98
50 untreated 0.25 69 38 0.5 13 13 1.0 13 nd control 18 75 untreated
0.25 nd 0.5 0 1.0 0 0 60 95 0.25 38 0.5 21 1 18 100 g fr. w. shoot
were shaken for ten minutes in 1 L of NH.sub.4OH. Plant material
discarded and the extract was sprayed onto the crop plants. nd =
not done.
Example 5
[0042]
6 Activity of aqueous or acetone extracts made from dry crushed
shoots of Inula against fungal diseases of crop plants. % Diseased
leaf area downy powdery late blight mildew in mildew in powdery in
potato cucumber cucumber mildew in solvent 4d 6d 6d wheat, 8d
untreated control 83 88 88 88 water 18 5 75 63 KOH 0.1N 88 4 75 63
NaOH 0.1N 63 28 50 25 NH.sub.4OH 1.4% 1 8 50 13 NH.sub.4OH 7% 3 4
25 15 (NH.sub.4).sub.2HPO.sub.4 0.3 0 50 3 0.1M (1.3%)
K.sub.2HPO.sub.4 0.1M 23* 3 88 20 Acetone 8 3 25 5 1 g dry crushed
shoots were shaken for thirty minutes in 50 ml of the solvent,
screened (100.mu.) and sprayed onto the crop plants. Plants were
inoculated one day after spray. *phytotoxic
Example 6
[0043]
7 Activity of dry crushed Inula shoots extracted with ammonium
hydroxide and ammonium phosphate (dibasic) against fungal disease
in crop plants. % diseased leaf area pow- pow- dery dery late late
downy mildew mildew blight in blight in mildew in in in tomato
potato cucumber wheat melon* solvent, conc. pH 4 d 6 d 8 d 11 d 14
d None -- 88 100 100 100 180 (NH.sub.4).sub.2HPO.sub.4 7.9 25 14 16
18 150 1.32% (0.1M) NH.sub.4OH 0.7% 10.97 25 22 28 15 55 (0.1M)
(NH.sub.4).sub.2HPO.sub.2 + NH.sub.4OH, mixed, 0.1M 9 + 1 8.25 29 5
27 15 80 7.5 + 2.5 8.6 31 13 16 3 100 5 + 5 9.0 11 6 6 0 40 2.5 +
7.5 9.4 20 0 11 1 25 1 + 9 9.8 16 2 16 8 20 water 6.2 75 80 58 75
180 acetone -- 0 16 11 8 130 1 g crushed dry shoots were shaken for
five minutes in 50 ml of solvent, screened and sprayed. Inoculation
took place three hours after spray. *Number of fungal colonies per
plant. Extracts were used six days after preparation. The molarity
of NH.sub.4OH is based on molecular weight of 35.
Example 7
[0044]
8 Activity of crushed dry Inula shoots extracts made with aqueous
solutions of ethylamine and ethylenediamine against fungal diseases
of crop plants. Potato late blight cucumber downy solvent, conc.
and 4d mildew 6d pH % blighted leaf area lesions/plant control
untreated 94 125 ethylamine 0.7% 15 5 (pH 12.3) ethylenediamine
0.6% 50 4 (pH 11.95) 1 g dry crushed shoots of Inula were shaken
for five minutes in 50 ml of the aqueous solvent, screened and
sprayed onto the plants. Plants were inoculated one day after
spray.
Example 8
[0045]
9 Activity of alkaline extracts or acetone of fresh Inula shoots
against disease development in crop plants. % infected leaf area
late blight in tomato powdery mildews in solvent and conc. 6d wheat
11d untreated control 88 75 ethylamine, 1% 0 38 ethylenediamine,
0.9% 0 38 ammonium hydroxide, 1% 1 20 (0.25% NH.sub.3 in water)
2-amino-2-methylpropanole 25 nt (AMP) 0.9% acetone 0 23 25 g fresh
shoots of Inula were dipped for ten minutes in 250 ml of the
solvent, and the extracts sprayed onto crop plants. Inoculation
took place three hours after spray.
Example 9
[0046]
10 Sequential extraction of fresh Inula shoots with acetone and
ammonium hydroxide and the activity of the extract against plant
disease. % infected leaf area powdery late blight in mildews in
First solvent Second solvent tomato, 6d wheat, 11d untreated
control 88 75 NH.sub.4OH, 1% -- 1 20 acetone -- 0 23 NH.sub.4OH, 1%
acetone 1 25 acetone NH.sub.4OH, 1% 38 75 25 g fresh Inula shoots
were dipped for ten minutes in the first solvent. Then washed with
water, blotted dry and dipped for ten minutes in the second
solvent. Extracts were sprayed onto the crop plants before
inoculation.
Example 10
[0047]
11 Antimicrobial activity, in vitro, of alkaline extracts of fresh
Inula shoots. fungal colonies bacterial colonies extraction made
with per dish per dish no extract, control 75 30 ethylamine, 1% 7
30 ethylene diamine, 0.9% 10 40 ammonium hydroxide, 1% 12 15
acetone (positive control) 4 0 0.5 ml extract (see Example 8) was
pipetted onto a 9 cm diam petri dish containing 10 ml Potato
Dextrose Agar. Dishes were exposed to aerial contamination for ten
minutes. Dishes incubated at 25.degree. C. for four days until data
were collected.
Example 11
[0048]
12 Dose - dependent activity of alkaline extracts made from dry
crushed Inula shoots with 1:1 mixture (pH 9.0) of 0.1M (1.32%)
(NH.sub.4).sub.2HPO.sub.4 and 0.1M (0.7%) NH.sub.4OH, against late
blight in tomato. g dry shoots blighted leaf area per 50 ml solvent
%, 4 days untreated control 88 solvent control 88 0.1 75 0.2 63 0.4
25 0.6 25 0.8 18 1 14 2 3 0.1-2 g dry crushed shoots were shaken
for five minutes in 50 ml of the solvent mixture, screened and
sprayed. Inoculation took place three hours later.
Example 12
[0049]
13 Activity of aqueous alkaline extracts made from dry crushed
Inula shoots against late blight in tomato. % diseased leaf area
solvent Inula extract solvent control untreated control 81 --
piperazine hydrate O.05M 3 81 (l%) pH 113 piperazine hydrate 0.1M 3
81 (2%) pH 11.3 triethanolamine 0.1M 23 81 (1.59%) pH 10.3 1 g dry
crushed shoots was shaken in 50 ml of the solvent for one hour,
screened and sprayed. Plants were inoculated three hours after
spray and disease was recorded four days later.
Example 13
[0050]
14 The effect of concentration of piperazine hydrate on the
activity of Inula extracts against late blight in tomato and
potato. piperazine hydrate % diseased leaf area conc.,% tomato
potato 0 91 95 0.06 18 25 0.12 11 25 0.25 4 25 0.50 18 44 1.00 0 75
2.00 8 75 (=0.1 M) 1 g dry crushed Inula shoots was shaken for one
hour in 50 ml of piperazine solutions, screened and sprayed.
Inoculation was done three hours after spray and disease recorded
five days after inoculation.
Example 14
[0051]
15 Efficacy of tetraethylene or piperidine aqueous solutions on
extraction of antifungal components from dry shoots of Inula %
diseased leaf area Tomato late cucumber downy wheat powdery
Treatment blight, 4d mildew, 6d mildew, 10d none (control) 88 88 50
tetraethylene 1% 63 31 50 (pH 11.2) piperidine 1% 88 94 25 (pH
12.0) Inula in tetraethylene 25 47 50 1% Inula in piperidine 1% 25
1 13 1 g dry crushed Inula shoots was shaken for ten minutes in 50
ml of the solvent, screened and sprayed. Inoculation was done three
hours after spray.
Example 15
[0052]
16 Activity of aqueous alkaline or acetone extracts made from Inula
shoots against mildews in cucumber and wheat % diseased leaf area
downy mildew powdery mildew powdery mildew in cucumber in cucumber
in wheat solvent 6d 11d 8d untreated control 75 100 100 Trizma-base
.RTM. 20 13 3 (Sigma) 1% piperazine 18 38 38 hydrate 1% piperadine
1% 5 25 18 imidazole 1% 0 0 0 acetone 2 25 25 (positive control) 1
g dry crushed leaves of Inula was shaken for ten minutes in 50 ml
of the solvent, screened and sprayed. Inoculation was done three
hours after spray.
Example 16
[0053]
17 Efficacy of sec - butylamine in extracting antifungal components
from dry Inula shoots. lesions per plant % diseased leaf area sec -
butylamine late blight in late blight powdery mildew conc., %
potato 3d tomato 4d cucumber 13d wheat 7d 0 130 88 88 100 0.06 60
63 14 23 0.12 0 25 14 0 0.25 5 8 9 15 0.5 0 15 15 3 1.0 (pH 11.5) 2
3 9 5 1 g dry crushed Inula shoots was shaken for ten minutes in 50
ml of sec-butylamine, screened and sprayed. Inoculation was done
three hours after spray.
Example 17
[0054]
18 Efficacy of imidazole in extracting antifungal component from
dry Inula shoots. % diseased leaf area late blight late blight
powdery mildew Treatment tomato, 5d potato, 5d wheat, 8d none 81
100 69 Imidazole only 0.12% pH 9.55 95 50 63 0.25% pH 9.70 88 50 50
0.5% pH 9.86 69 25 23 1% pH 9.95 50 18 25 Inula extract in
imidazole 0.12% 30 5 0 0.25% 20 18 18 0.5% 30 30 0 1.0% 30 15 0
Inula in acetone 5 0 3 1 g of dry crushed Inula shoots was shaken
in 50 ml of imidazole for ten minutes, screened and sprayed.
Inoculation was done three hours after spray.
Example 18
[0055]
19 Composition of a 50 WP formulation of Inula shoots and its
activity against fungal plant diseases. Composition: 5 g dry Inula
shoot powder 2.5 g imidazole 2.5 g inert carriers and surfactants
(Sandoz) Activity % diseased leaf area g product per late blight
late blight powdery mildew 50 ml water pH tomato 5d potato 5d wheat
13d 0 -- 98 100 38 0.25 8.20 75 75 25 0.5 8.21 38 63 13 0.75 8.22
31 25 0 1 8.23 23 18 0 1.25 8.24 20 20 0 1.5 8.25 15 13 0 2 8.25 1
10 0
Example 19
[0056]
20 Composition of a 50 WP formulation of Inula shoots and its
activity against fungal plant diseases. Composition: 5 g dry Inula
shoot powder 3.75 g imidazole 1.25 g inert carriers and surfactants
(Sandoz) Activity % diseased leaf area g product per late blight
late blight powdery mildew 50 ml water pH tomato 4d potato 5d wheat
9d 0 -- 95 81 38 0.25 8.28 75 69 25 0.5 8.29 63 25 0 0.75 8.32 38
15 0 1 8.33 25 13 0 1.25 8.35 21 13 0 1.5 8.36 13 18 0 2 8.37 6 8
0
Example 20
[0057]
21 Composition of 45 WP formulation of Inula shoots and its
activity against fungal plant diseases. Composition: 5 g dry Inula
shoot powder 5 g imidazole 1 g inert carriers and surfactants
(Sandoz) Activity % diseased leaf area g product per 50 ml late
blight in water pH tomatoes 4d 0 -- 75 0.25 8.18 75 0.5 8.23 50
0.75 8.28 50 1 8.29 38 1.5 8.30 25 2 8.38 3
Example 21
[0058]
22 Composition of 50 WP formulation of Inula shoots and its
activity against fungal plant diseases. Composition: 5 g dry Inula
shoot powder 2.5 g dibasic ammonoim phosphate 2.5 g Avicel PH - 101
.RTM. (Fluka) = crystalline cellulose % diseased leaf area Activity
downy g product late blight mildew powdery per 50 ml late blight in
in potato, in cucumber, mildew in water pH tomato, 4 d 6 d 6 d
wheat, 9 d 0 -- 98 98 50 75 0.1 7.33 75 93 13 75 0.2 7.44 75 93 8
50 0.4 7.46 63 38 3 50 0.6 7.46 63 30 1 25 0.8 7.46 18 23 3 23 1
7.46 23 20 0 20 1.5 7.46 23 18 0 18
Example 22
[0059]
23 Composition of 50 WP formulation of Inula shoots and its
activity against fungal plant diseases. Composition: 5 g dry Inula
shoot powder 2.5 g Trizma - Base .RTM. (Sigma) 2.5 g Avicel PH -
101 .RTM. (Fluka) Activity % diseased leaf area g product per late
blight late blight in powdery mildew 50 ml water pH tomato, 3d
potato, 5d in wheat, 8d 0 -- 95 95 75 0.2 8.55 88 95 50 0.4 8.62 88
93 50 0.6 8.67 38 69 25 0.8 8.68 23 38 25 1 8.70 30 25 25 1.5 8.72
23 25 25 2 8.74 25 25 25
Example 23
[0060]
24 Composition of 50 WP formulation of Inula shoots and its
activity against fungal plant diseases. Composition: 5 g dry Inula
shoot powder 5 g dibasic ammonium phosphate Activity % diseased
leaf area g product late blight in potato per 50 ml late blight in
lesions/plant powdery mildew water pH tomato, 4 d 3 d 4 d in wheat,
7 d 0 -- 95 >500 88 88 0.2 7.42 38 200 69 38 0.4 7.45 25 180 69
38 0.6 7.45 23 50 25 23 0.8 7.45 20 40 25 13 1 7.46 3 20 20 13 2
7.47 1 5 8 3
Example 24
[0061]
25EXAMPLE 24 Composition of alkaline wettable powder formulations
of Inula used to control plant diseases. Percent Other Formula a-i
Alkaline agent pH components Inula leaves (27%) 25 none 6.8 Zeofree
80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%) 25
amines (10%) 7.9 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779
(10%) Inula leaves (25%) 12.5 triethanolamines 8.5 Zeofree 80 (95%)
Stepsperse DF500 (10%) Monoamine 779 (20%) Triethanolamine (50%)
Inula leaves (25%) 25 (NH.sub.4).sub.2HPO.sub.4 (25%) 7.5 Zeofree
80 (25%) diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%)
Inula leaves (25%) 25 (NH.sub.4).sub.2HPO.sub.4 (25%) 8 Hubersorb
600 (25%) diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%)
Inula leaves (25%) 25 (NH.sub.4).sub.2HPO.sub.4(25%) 8 Hubersorb
600 (40%) diammonium phosphate (25%) Stepan DF 500 (10%) Inula
leaves (25%) 25 (NH.sub.4).sub.2HPO.sub.4 (25%) 8 Hubersorb 600
(24%) diammonium phosphate (25%) Stepan DF 500 (5%) 40% Tdet 09 on
Zeolex 7A (5%)
Example 25
[0062]
26EXAMPLE 25 Late blight in potato. % infected leaf area 6 days
Conc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 100 100
100 100 Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula
leaves (25%) 50 25 38 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779
(10%) Inula leaves (25%) 25 25 18 Zeofree 80 (95%) Stepsperse DF500
(10%) Monoamine 779 (20%) Triethanolamine (50%) Inula leaves (25%)
100 69 50 Zeofree 80 (25%) diammonium phosphate (25%) Reax M100
(5%) Kaolin (20%) Inula leaves (25%) 100 50 20 Hubersorb 600 (25%)
diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves
(25%) 50 20 0 Hubersorb 600 (40%) diammonium phosphate (25%) Stepan
DF 500 (10%) Inula leaves (25%) 100 88 25 Hubersorb 600 (24%)
diammonium phosphate (25%) Stepan DF 500 (5%) 40% Tdet 09 on Zeolex
7A (5%)
Example 26
[0063]
27EXAMPLE 26 Late blight in tomato. Suspensions were used
.about.100 hours after preparation. % infected leaf area 4 days
Conc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 95 88 75
38 Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves
(25%) 19 15 8 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%)
Inula leaves (25%) 18 1 0 Zeofree 80 (95%) Stepsperse DF500 (10%)
Monoamine 779 (20%) Triethanolamine (50%) Inula leaves (25%) 75 63
25 Zeofree 80 (25%) diammonium phosphate (25%) Reax M100 (5%)
Kaolin (20%) Inula leaves (25%) 75 63 25 Hubersorb 600 (25%)
diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves
(25%) 50 38 9 Hubersorb 600 (40%) diammonium phosphate (25%) Stepan
DF 500 (10%) Inula leaves (25%) 75 38 19 Hubersorb 600 (24%)
diammonium phosphate (25%) Stepan DF 500 (5%) 40% Tdet 09 on Zeolex
7A (5%)
Example 27
[0064]
28EXAMPLE 27 Downy mildew in cucumber. % infected leaf area 6 days
Conc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 100 63 38
38 Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves
(25%) 38 38 10 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%)
Inula leaves (25%) 20 20 13 Zeofree 80 (95%) Stepsperse DF500 (10%)
Monoamine 779 (20%) Triethanolamine (50%) Inula leaves (25%) 50 30
30 Zeofree 80 (25%) diammonium phosphate (25%) Reax M100 (5%)
Kaolin (20%) Inula leaves (25%) 44 13 13 Hubersorb 600 (25%)
diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves
(25%) 44 8 8 Hubersorb 600 (40%) diammonium phosphate (25%) Stepan
DF 500 (10%) Inula leaves (25%) 75 63 44 Hubersorb 600 (24%)
diammonium phosphate (25%) Stepan DF 500 (5%) 40% Tdet 09 on Zeolex
7A (5%)
Example 28
[0065]
29EXAMPLE 28 Downy mildew in grapes. % sporulating leaf area 8 days
Conc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 97 60 5 0
Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves (25%)
25 25 20 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%) Inula
leaves (25%) 50 25 15 Zeofree 80 (95%) Stepsperse DF500 (10%)
Monoamine 779 (20%) Triethanolamine (50%) Inula leaves (25%) 70 15
0 Zeofree 80 (25%) diammonium phosphate (25%) Reax M100 (5%) Kaolin
(20%) Inula leaves (25%) 60 20 20 Hubersorb 600 (25%) diammonium
phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves (25%) 50
25 20 Hubersorb 600 (40%) diammonium phosphate (25%) Stepan DF 500
(10%) Inula leaves (25%) 80 80 50 Hubersorb 600 (24%) diammonium
phosphate (25%) Stepan DF 500 (5%) 40% Tdet 09 on Zeolex 7A
(5%)
Example 29
[0066]
30EXAMPLE 29 Powdery mildew in wheat. % infected leaf area 8 days
Conc. % a-i Formulation 0 0.25 0.5 1.0 Inula leaves (27%) 50 50 25
13 Zeofree 80 (60%) Polyfon F (3%) Synfactant (10%) Inula leaves
(25%) 25 13 0 Zeofree 80 (62%) Polyfon F (3%) Monoamine 779 (10%)
Inula leaves (25%) 25 13 0 Zeofree 80 (95%) Stepsperse DF500 (10%)
Monoamine 779 (20%) Triethanolamine (50%) Inula leaves (25%) 38 19
13 Zeofree 80 (25%) diammonium phosphate (25%) Reax M100 (5%)
Kaolin (20%) Inula leaves (25%) 38 25 13 Hubersorb 600 (25%)
diammonium phosphate (25%) Reax M100 (5%) Kaolin (20%) Inula leaves
(25%) 30 25 0 Hubersorb 600 (40%) diammonium phosphate (25%) Stepan
DF 500 (10%) Inula leaves (25%) 25 25 5 Hubersorb 600 (24%)
diammonium phosphate (25%) Stepan DF 500 (5%) 40% Tdet 09 on Zeolex
7A (5%)
[0067] In view of the teachings presented herein, other
modifications and variations of the present inventions will be
readily apparent to those of skill in the art. The foregoing
discussion, and description are illustrative of some embodiments of
the present invention, but are not meant to be limitations on the
practice thereof. It is the following claims, including all
equivalents, which define the scope of the invention.
[0068] Any patents or publications mentioned in this specification
are indicative of the levels of those skilled in the art to which
the invention pertains. These patents and publications are herein
incorporated by reference to the same extent as if each individual
publication was specifically and individually indicated to be
incorporated by reference.
[0069] One skilled in the art will readily appreciate that the
present invention is well adapted to carry out the objects and
obtain the ends and advantages mentioned, as well as those inherent
therein. The present examples along with the methods, procedures,
treatments, and specific compositions described herein are
presently representative of preferred embodiments, are exemplary,
and are not intended as limitations on the scope of the invention.
Changes therein and other uses will occur to those skilled in the
art which are encompassed within the spirit of the invention as
defined by the scope of the claims.
* * * * *