[COPY]
United States Patent [19]
H. E. Kistner, Sr.
[54] FERTILIZER UTILIZING A
CHlTIN-PRODUCING FUNGUS AND
METHOD FOR ITS USE
[76] Inventor: Harold E. Kistner, Sr., 1500 W.
Matlock, #22, Aransas Pass TX ,
78336
[21] Appl. No. : 764,932
[22] Filed: Aug. 12, 1985
[51] Int, cl),4 ....... CO5F 11/08;
[52] U.S. CL .. .. .. 71/7;435/918;
[58] Field of Search ...... .. 426/331,634,807;
47/57.6, DIG. 9, DIG. 10; 71/1, 6, 7; 435/171,
915, 918; 424/115
Patent Number: 4,670,037
Date of Patent: Jun. 2, 1987
References Cited
[56]
US. PATENT DOCUMENTS
3,115,404 12/1963 Carney ....71/6
Primary examiner---Ferris H. Lander
Attorney, Agent, or Firm-Pearne, Gordon, McCoy &
Granger
[57] ABSTRACT A novel natural fertilizer and method of
its use are Provided. The fertilizer acts to increase
the Yield of food crop producing plants. The fertilizer
comprises less than 0.05% of an active agent in a stable
carrier that is non-toxic to the plants and the active
agent. The active agent is a fungus of the genus
Aspergillus. The fertilizer can be used by coating the
seeds of the food crop producing plant at or prior to
the time of planting by dusting the ends of a cut plant,
by dusting the roots of a seedling, or by treating the
soil in which the plant will be grown.
6 Claims,
No Drawings
FERTILIZER UTILIZING A CHITIN-PRODUCING FUNGUS AND
METHOD FOR ITS USE BACKGROUND OF THE INVENTION
The invention relates to fertilizers and plant growth
and yield promoters, and to processes for the use of
these products.
Fertilizers and plant growth and yield promoters have
long been used to increase plant growth and crop yield.
While mankind first used natural products, such as waste
products, as fertilizers, the use of synthesized
fertilizers has become increasingly prevalent. This
proliferation of synthetic fertilizers has given rise to
concern for the health of individuals and ecological
systems exposed to these products and to the by-products
from their production.
The present invention responds to these concerns by
providing a fertilizer having an active agent which
occurs naturally, which has been shown causes
significantcant increases in plant growth and crop
yields, and which is relatively economical and
convenient to use. The active agent is a fungus of the
genus Aspergillus.
DISCUSSION OF THE PREVIOUS ART Naturally occurring
products have long been used as fertilizers and plant
growth promoters. Such products have included manure,
fish products, and crop residues.
Another example of naturally occurring products that are
used in agricultural science are certain strains of
bacteria. It is understood that the addition of
bacteria such as Rhizobia and Azotobacter can aid plants
in the breakdown of soil materials to form plant
nutrients, as well as in the in fixation of atmospheric
nitrogen. An example of a teaching illustrating this is
found in US. Pat.No.4,061,488 to Mann, which teaches
methods of using a plant treating mixture incorporating
spores of Bacillus uniflagellatus.
It has been recognized as well that certain types of
fungus or their products can be used by agronomists.
US. Pat. No. 4,370,351 relates to a method of
producing a food or feed product, such as a cereal
grain, which has been treated with a spawn culture of
Pleurotus to increase the protein content of the food.
It is also known that the genus of fungi used in the
present invention, Aspergillus, can be useful for
agricultural purposes. US. Pat. No. 3,043,748,
having the same inventor as the present invention,
discloses a product resulting from the fermentation of
sucrose by Aspergillus us flavus, oryzae. which serves
as a growth-promoting substance.
OBJECTS AND SUMMARY OF THE
INVENTION
It is an object Of the present invention to provide a
new fertilizer or plant yield and growth promoter par-
ticularly useful for commercial plants such as commer-
cial crop producers.
It is another object of the invention to provide methods
for the use of the new fertilizer.
The fertilizer of the present invention concerns the use
of fungus as the active agent. It is believed that all
chitin-producing fungi may be used in accordance with
the invention as a fertilizer. However, the strains of
fungi which have been demonstrated to be useful are from
the Euascomycetae subclass of Asomycetes, and carriers
are finely ground calcium carbonate, talc, and silt or
the material washed from sand. An example of a material
which has been found to be unsuitable as a carrier is
wheat middlings. It is believed that this substance
results in agglutination by the mold.
The fertilizer should contain 5 to 500 parts per million
parts (i.e., 0.0005% to 0.05%) of active agent, and
preferably 5 to 100 parts per million (i.e., 0.0005%.
to 0.01%), and most preferably 10 to 50 parts of active
agent per million (i.e., 0.001% to 0.005%). Other plant
growth additives which do not interfere with the
intended tended activity of the active agent may be
added. Trace elements would be an example of such
additives.
The fertilizer may be used in various ways. Seeds may
be coated at planting time or before planting time with
sufficient fertilizer to coat the seed. The exact
amount of fertilizer necessary to coat the seed will
vary with the size of the seed. Smaller seeds will
require more, since they have a greater surface area.
One-half 20 cup of growth-promoter is sufficient to coat
one bushel of seed corn.
It is also possible to dust the cut ends of plant
cuttings before planting. Plants that may be planted in
this manner ner include agricultural crops such as
potatoes, or even house plants that are grown in this
manner.
The growth-promoter may be dusted on the roots of
seedlings which will be transplanted, such as tomatoes
or cabbage. The growth-promoter may be applied, as by
dusting the tilled field, to the soil prior to planting.
The following examples demonstrate the products, use,
and effectiveness of the growth-promoter of the present
invention.
PART 2 KISTNER CHITIN
PATENT
EXAMPLE 1
Production of the active agent for the fertilizer of the
present invention can be accomplished as follows. A
culture of Aspergillus flavus-oryzae, NRRL 458, is
isolated by use of Czapek-Dox medium with agar. This
strain of fungi is on deposit at the Northern
Utilization Branch at Peoria, I11. The Czapek-Dox
medium consists of NaNo3, 3.0 grams; K2HP04, 1 .0 gram;
MgS04.7- H20, 0.5 gram; KC1, 0.5 gram; FeS04.7H20, 0.01
gram; sucrose 30.0 grams; and agar 15.0 grams in 1 liter
distilled water. All components are of reagent grade.
This medium is sterilized at 15 pounds pressure for 15
minutes. The medium is inoculated with spores of the
mold culture and is subsequently incubated at 25 degrees
C. for two to three days. Single colonies of the
culture will develop velop and isolated mold spores can
be selected by a trained observer.
The culture material that acts as the active agent can
be prepared from the isolated mold spores by either of
two methods The culture material may he grown in
sterilized, distilled water that is aerated with air and
carbon dioxide gas, or the culture material may be grown
in any suitable carbohydrate medium, such as wheat bran.
To cause growth in water, isolated mold spores are
introduced to sterilized, de ionized distilled water in
a flask by air floatation of spores into the water Air
and carbon dioxide are bubbled through the water and
mold mixture. The mixture is incubated at 28 degrees C.
until a sufficient quantity of culture material has
grown. The culture material is separated from the water
by filtration, decanting or evaporation. This is used
as the active agent.
For the second growth method, isolated mold spores are
air-floated into a 500 milliliter flask containing 32
grams of wheat bran which has been moistened with 16
milliliters of 0. 1 normal hydrochloric acid. The
inoculated bran is incubated at 20" to 25" C. The
flasks should be left upright, until rapid growth is
completed, specifically when the material will remain
caked in the bottom when laid flat. This material is
allowed to dry. The dried material is used as the
active agent. It consists of fungus and any residual
bran.
In order to assure that the proper amount of active
agent is added to the carrier for the maximum
efficiency, a sample of the active agent is tested.
This standardization procedure can be accomplished by
the fermentation of starch. Fifty-six grams of finely
ground corn is placed in a 500 milliliter flask to which
250 milliliters of distilled water is added. This
mixture is autoclaved at 15 pounds of pressure for 15
minutes. The mixture is allowed to cool to 61 degrees
C. and one gram of mold spore culture is added to the
mixture. This is cultured with yeast at 28 degrees C.
This temperature is maintained tained until the growth
medium is spent. The weight loss is measured every 12
hours.
In order to determine the glucose which is produced by
the enzyme activity of the mold spore culture, a control
is also run which was not treated with mold spore
culture. The weight loss for the control represents the
amount of fermentable sugar native to the corn. This
value is subtracted from the weight loss for the treated
sample. The weight of the carbon dioxide produced
equals 95.5% of the value of the alcohol produced. The
weight of the alcohol produced equals 51.1 % of the
value of the glucose fermented. Thus, the weight of the
carbon dioxide produced equals 48.9% of the glucose
fermented. It has been determined that 20 to 30 parts
per million of active agent in the carrier is the
optimal level when the sample which has been
standardized results in the production of 12-14 grams of
carbon dioxide over a 60-hour period.
EXAMPLE 2
This and the following examples illustrate methods of
using the invention, and demonstrate the efficacy of the
present invention. Active agent was isolated and
prepared by growth in wheat bran as described above.
The active agent was mixed, 25 parts per million active
agent in a horizontal mixer to form a substantially
uniform disbursement with a carrier of fine silt which
was washed from alluvial deposit to form the fertilizer
of the present invention. This carrier is also referred
to as sand washings. A field test was conducted
comparing the crop yield and the root growth of Lincoln
pea plants grown from treated seeds with those of
Lincoln pea plants grown from non-treated seeds. The
seeds were treated with the fertilizer by coating the
seeds. Two 13-foot rows of treated seed were planted
intermediate two 13-foot rows of untreated seed used as
a control. Rows I and IV were the control rows.
Twenty-four inch netting was placed over the seeds to
protect the plants from pests. The peas were allowed to
mature and the peas were harvested every other day,
three times. The yield is shown below.
Yield in Pounds
Control Treated Seeds
Row I Row IV Row II Row 111
First Harvest 0.90 1.40 1.30 1.70
Second Harvest 0.25 0.75 0.85 0.50
Third Harvest 0.00 0.10 0.00 0.20
1.15 2.25 2.15 2.40
The total yield for the control was 3.40 pounds, while
the total yield for the treated seeds was 4.55 pounds.
This represents a 33.8% increase in yield for the
treated plants. At the bloom stage and after harvest, a
number of plants were carefully removed from the soil.
The dirt was carefully cleaned from the roots so that
the root system and the nodules on the root system were
not disturbed. The roots were dried and mounted for
com parison of the root system and the nodules.
Nodules are round swellings on a root which are
inhabited by symbiotic bacteria. These bacteria help
the plant to assimilate needed nutrients, most notably
nitrogen. Thus, an in- creased nodule count indicates a
greater ability to as- similate nitrogen. This in turn
contributes to greater crop yield. Comparison of the
roots of the treated and the con- trol plants both at
the bloom stage and the mature stage shows a greater
root development and greater development of nodules in
the treated plant.
EXAMPLES 3-5
Similar field tests were conducted for red beets, car-
rots, and popcorn. The tables below represent the per-
centage weight increase for the plants treated with the
fertilizer at 25 ppm Aspergillus flavus-oryzae active
agent in sand washings as compared to an untreated
control grown at the same period in the same soil
conditions in a neighboring plot.
Percentage Yield Increase
Plant (by weight)
Red beets 50.0%
Carrots 37.5%
Popcorn 33.3%
Comparative photographs of the resultant crops
illustrate that the individual vegetables yielded from
the treated plants were generally larger than the
vegetables of the control plants.
EXAMPLE 6
A similar field test was conducted for Pontiac potatoes
The test crop was produced from cuttings dusted at the
fresh cut end with a fertilizer having 25 ppm
Aspergillus Flavus oryzae active agent in sand washings.
A control crop was grown in a neighboring plot from
untreated cuttings. The treated plants yielded 18.8% by
weight more potatoes than the control plants. Once
again, comparative photographs illustrate that the
treated plants yielded a generally higher quality
product than the control plants. .
EXAMPLE 7
A similar field test was conducted for Tendergreen
beans. The test crop was grown from seeds coated with
25 ppm Aspergillus flavus oryzae active agent in sand
washings. The control crop was grown at the same period
in a neighboring plot. After several weeks, young
plants were removed from the ground and the dirt was
carefully removed from the root system. Care was taken
not to disturb root hairs and nodules on the roots. The
roots were carefully dried and mounted far comparison.
A comparison between the estimated num- ber of visible
nodules per plant on test plants and on treated plants
is seen below.
IMMATURE PLANTS
Plain Nodule Treated Nodule
Plants Count Plants Count
Control 1 48-58 Test 1 56-66
Control 2 6-16 Test 2 18-28
Control 3 16-26 Test 3 36-46
About four weeks later, mature plants were similarly
removed from the ground. The dirt was carefully removed
from the root system. The roots were dried and mounted
for comparison. Because of more extensive root system,
it was difficult to estimate the number of nodules for
the plants; however, the treated plant ap- peared to
have significantly more nodules than the con- trol and,
further, to have a more extensive root system.
EXAMPLE 8
A similar field test was conducted for Great Northern
Beans. Test plants were grown from seed dusted with
fertilizer of the invention containing 25 ppm of
Aspergil- lus ,flavus oryzae active agent in sand
washings. The control plants were grown from untreated
seed in a neighboring plot under the same conditions as
the test plants. The treated plants yielded a 33.3%
increase by weight in the yield. After harvest, plants
were carefully removed from the soil and the dirt was
removed from the root system for a comparison of the
roots of treated versus untreated plants, as in Examples
1 and 7. The treated plant demonstrated a significant
increase in the root growth, as well as in the number of
nodules present.
EXAMPLE 9
A laboratory test was conducted comparing germination of
hybrid seed corn coated with the fertilizer of the
present invention at varying levels of active agent.
The corn was allowed to germinate in filter paper, kept
moist with sterilized distilled water. The test was
conducted with plain corn and with corn treated with
Aspergillus flavus,oryzae active agent at 20 ppm, 25
ppm, and 30 ppm in sand washings. After two weeks of
growth, the seedlings were mounted on graph paper and
allowed to dry. Measurements were taken of the root and
stalk growth for ten seedlings for each group. The
results are shown below. The measurements are in
centimeters +0.4 centimeters.
Control 20 ppm 25 ppm 30 ppm
Stalk/Root Stalk/Root Stalk/Root Stalk/Root
Kernel Growth Growth Growth Growth
1 4.8/1.8 5.0/1.2 5.0/13.2 5.0/5.6
2 4.4/2.0 4.6/6.4 5.8/6.4 4.6/2.4
3 4.6/0.2 4.2/8.6 5.4/7.0 3.6/2.0
4 3.6/3.8 3.6/2.0 5.6/3.0 2.0/3.4
5 2.8/3.4 3.2/6.0 5.2/8.4 4.2/4.8
6 2.2/0.2 3.4/4.0 4.4/8.0 3.2/2.4
7 2.2/0.4 3.0/5.6 4.5/9.4 3.2/4.2
8 2.8/3.0 2.4/2.0 4.0/7.4 2.5/0
9 ------ 1.6/0 4.0/7.4 1.2/1.8
10 ------ 1.2/2.3 2.4/6.4 1.5/2.8
The increase in root and stalk growth for the treated
seeds was striking. The test demonstrates that the
optimal level of active agent was 25 ppm. The foregoing
example Show that the fertilizer in accordance with the
invention is an effective fertilizer and plant growth
and yield promoter. Although the preferred embodiment
of this invention has been shown and described, it
should be understood that various modifications may be
reverted to without departing from the scope of the
invention as disclosed and claimed herein.
What is claimed is:
1. A fertilizer which produces increased yields of a
food crop producing plant, which comprises less than
1.05% of an active agent consisting of a fungus of the
species Aspergillus Flavus Oryzae which can be grown at
temperatures up to 30' C. and possesses the ability to
fix nitrogen and to incorporate it into chitin, the
active agent being substantially uniformly dispersed in
a carrier that is stable during storage and non-toxic to
the plant wherein the carrier is chosen from the group
consisting of finely ground calcium carbonate, talc,
sand, sand washings, and silt. 2. A method for
increasing the yield of a food crop yielding plant which
comprises treating the food crop yielding plant with an
effective amount of fertilizer to cause an increase in
the food crop yield, the fertilizer comprising less than
0.05% of an active agent consisting of a fungus of the
species Aspergillus Flavus Oryzae which can be grown at
temperatures up to 30 degrees C. and posses the ability
to fix nitrogen and to incorporate it into chitin, the
active agent being substantially uniformly dispersed in
a carrier that is stable during storage and non-toxic to
the plant 3. A method in accordance with claim 2,
wherein the plant is grown from seed, and the seed is
coated with the fertilizer. 4. A method in accordance
with claim 2, wherein the plant is grown from a cutting
having a cut end and the cut end of the cutting is
dusted with the fertilizer. 5. A method in accordance
with claim 2, wherein the plant is grown from a seedling
having roots, and the fertilizer is dusted on the roots
of the seedling. 6. A method for increasing the yield
of a food crop yielding plant which comprises treating
the soil in which the food crop yielding plant is grown
with a fertilizer, the fertilizer comprising less than
0.5% of an active agent consisting of a fungus of the
species Aspergillus Flavus oryzae which can be grown at
temperatures up to 30 degrees C. and posses he ability
to fix nitrogen and to incorporate it into chitin, the
active agent being substantially uniformly dispersed in
a carrier that is stable during storage and non-toxic to
the plant.