Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
CA 02841043 2014-01-06
WO 2012/143684 PCT/GB2012/000366
1
COMPOSITIONS FOR GROWTH AND VIGOUR FROM OILSEEDS
The present invention relates to coating compositions including an organic
component for
applying to seeds of oilseed plant from which roots and shoots are capable of
growing, uses
of coating compositions on oilseed plant seeds, methods of producing such
coating
compositions and oilseed plant seed coated with such coating compositions. In
particular,
the invention relates to coating compositions that comprise an organic
material that provides
protection from environmental stresses to oilseed plant seed.
Young oilseed plants grown from oilseed plant seed are vulnerable to abiotic
and
environmental stresses, particularly in growing habitats that have low
rainfall and/or sub-
optimal soil quality. Losses due to sub-optimal soil quality are typically
realised in the growth
of young plants lacking plant vigour in which the plants do not become well
established, such
as where the rooting systems do not develop and in circumstances where
essential
elements in the soil are not readily available. Agronomic losses due to young
oilseed plants
not being well established remain unacceptably high on soils which are for
example mineral
deficient despite the employment of conventional inorganic oilseed plant seed
coatings that
typically include essential elements for establishing young seedlings. A
problem with the use
of such conventional coatings is that they introduce nutrients to the soil in
unbalanced
quantities and this can have adverse effects on plant growth and vigour in
unforeseen ways.
Additionally, such conventional coatings are typically applied in the form of
wet slurries to
oilseed plant seeds. Once applied, the coatings are typically dried on the
seeds and this
drying may cause further abiotic stresses, which in turn may have deleterious
consequences
on the viability of young plants grown therefrom. Additionally, such
conventionally applied
coatings may not be applied to oilseed plant seeds evenly, and as a
consequence, such
coatings tend to be susceptible to chipping and/or flaking. Furthermore, the
degree of
coating uniformity of such conventionally applied coatings typically is not
optimal, with a
percentage of seeds of any one batch receiving little or no coating depending
on the coating
method being deployed.
In the following description, the terms "oilseed treatment" and "oilseed
coating" are used
interchangeably for the compositions of the invention and their uses to treat
oilseed plant
seeds, by any of the specific methods described in the prior art that provide
an improvement,
typically an enhancement, of seedling vigour. The commonly used ingredients in
oilseed
=
plant seed treatment compositions (sometimes designated as formulations)
include antidotes
and safeners; fertilisers, micronutrients and inoculants; bioregulators of
natural or synthetic
origin which are either hormones or interfere in hormone metabolism and do not
influence
plant nutrition; and/or bioregulators which interfere with plant growth by
enhancing nutrient
uptake.
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
2
It has now been found that by using certain organic materials as components of
coatings on
oilseed plant seeds, together with the application of inorganic components
and/or biological
agents, plant vigour and plant growth of plant seedlings grown from oilseed
plant seeds is
improved relative to the plant vigour and plant growth of seedlings grown from
conventionally
treated seeds. It has further been found that the quantity of additives,
particularly inorganic
fertilisers that is required per unit of oilseed plant seed weight is less
than that required using
conventional farming techniques.
It is an object of the present invention to supply improved oilseed plant seed
coatings
comprising organic components for oilseed plant seeds.
It is a further object of the present invention to provide improved oilseed
plant seed coatings
comprising a minimum amount of additives.
These and other objects of the invention will become apparent from the
following description
and examples.
According to the present invention there is provided an oilseed coating
composition in the
form of organic material in particulate form that comprises
i) at least one organic material selected from waxes having a melting point of
?_50 Centigrade; and
ii) at least one additive for enhancing seedling vigour and/or seedling growth
from oilseed
plant seed wherein the at least one additive is selected from one or more
inorganic additives
and/or one or more live biological agents.
The organic materials of use in the invention act as a carrier for desired
additives for placing
on or near to seeds.
For the purposes of the present invention an "oilseed plant seed" is one from
which roots
and shoots are able to grow. Reference to "oilseed plant seed" and "oilseed
plant seeds" is
used interchangeably herein and means seeds, typically viable seeds, to which
compositions
of the invention may be applied. Furthermore, "oilseed plant seed" and
"oilseed plant seeds"
as provided herein means seeds that are capable of germinating to at least
conventional
levels of germination typical of the relevant oilseed plant species under
consideration.
"oilseed plants" for the purposes of the present invention are ones which are
recognised as
such by the skilled addressee. Oilseed plant seeds suitable for coating with
compositions of
the invention include those that may be used for the planting of oilseed
plants include those
selected from members of the Crucifer family (Canola (B. campestris) and
oilseed rape (B.
napus), sunflower, peanut, safflower, sesame, nut oils, carob, coriander,
mustard, grape,
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
3
linseed (flax), dika, hemp, okra, pine, poppy, castor, jojoba and the like.
Generally, the particles of use in seed coating compositions of the invention
possess a
volume mean diameter of a certain size as defined herein. To obtain particles
of organic
materials of a volume mean diameter applicable for use in the invention,
organic materials in
the form of, for example, 1 to 5 kilogram blocks or tablets may be broken up
or kibbled into
small millimetre-sized pieces (such as from 2mm ¨ 8mm approximate diameter in
size, for
example from 4mm to 6mm) in a kibbling machine. The millimetre-sized pieces
can then be
passed through a comminuting means such as a standard mill, e.g. an Apex
Comminuting
mill, and milled or comminuted into particles having an approximate diameter
in the range
from 100pm - 500pm, for example from 250pm - 300pm. The micron-sized
comminuted
particles can then be passed through a micronising apparatus, such as an AFG
micronising
air mill to obtain particles of a desired VMD range, such as from 15pm - 20pm,
that is of use
in the present invention. The skilled addressee will appreciate that such
procedures for
obtaining small particles are well known in the art. Preferably, dry powder
compositions of
the invention comprise composite particles having a volume mean diameter of al
Opm, for
example of 10pm, 11pm, 12pm, 13pm, 14pm, 15pm up to 40pm or any value
thereinbetween. As stated herein, the volume mean diameter of the composite
particles is
typically .?_10pm or ?..12pm and may lie in the range from 10pm to 200pm and
may have a
value that lies anywhere there inbetween, for example from .10pm to 100pm; or
from
?_10pm to 40pm; or from .-10pm to 30pm or any desired volume mean diameter
value in
between. Preferably, dry powder compositions of the invention comprise
particles having a
volume mean diameter of ?.10pm, for example of 10pm, 11pm, 12pm, 13pm, 14pm,
15pm
and the like up to any volume mean diameter of choice, such as up to 200pm or
any volume
mean diameter in between for example 40pm or 30pm. Such compositions are
considered to
be less of a thoracic hazard to humans and are not thought to be allergenic.
The organic material used in the present invention is selected from organic
materials that
can be applied to oilseed plant seeds either as a powder wherein the powder
particles are of
a pre-determined volume mean diameter (VMD) or the powder particles are
applied in liquid
form, such as an oleaginous formulation or as an aqueous formulation. In
liquid formulations,
particles of a pre-determined volume mean diameter are suspended therein in a
suspension
formulation and applied to oilseed plant seeds, which are then dried using
conventional
drying procedures. Preferably, the organic material is applied to oilseed
plant seeds in a dry
powder form. Such organic materials include additives as herein defined and
may include
added further components such as added UV blockers or added antioxidants or
the like. Dry
powders of the present invention may be made up of one or more organic
materials that
have a melting point at or above 50 C and which are of use in the present
invention.
Suitable organic materials of use in the invention include waxes having a
melting point of
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
4
?50 C, more preferably of ?_60 C, and most preferably are made up of hard
waxes having a
melting point of ?-70 C. Suitable organic materials of use in the invention
include waxes
selected from natural, synthetic and mineral waxes. Examples of natural waxes
of use in the
present invention include carnauba wax, beeswax, montan wax, Chinese wax,
shellac wax,
spermaceti wax, myricyl palmitate, cetyl palmitate, candelilla wax, castor
wax, ouricury wax,
sugar cane wax, retamo wax, rice bran wax and the like. In a preferment, the
organic
material is selected from carnauba wax, beeswax, montan wax, Chinese wax,
shellac wax,
spermaceti wax, myricyl palmitate, cetyl palmitate, candelilla wax, castor
wax, ouricury wax,
and rice bran wax; or a mixture of two or more thereof.
Synthetic waxes of use in the present invention include suitable waxes
selected from paraffin
wax, microcrystalline wax, Polyethylene waxes, Fischer-Tropsch waxes,
substituted amide
waxes, polymerized a-olefins and the like.
Mineral waxes of use in the invention include montan wax (e.g. Lumax0 Bayer)
ceresin wax,
ozocerite, peat wax and the like.
Waxes of use in the invention typically display a high enthalpy of lattice
energy during melt.
Preferably the organic material is carnauba wax which may be applied in liquid
form,
typically in the form of a suspension, or powder form as discrete particles.
Preferably, the
organic material is applied in dry powder form to oilseed plant seeds.
Generally, the particles
of use in the invention possess a volume mean diameter of a1Opm, such as
.?_12pm such as
in the range of from ?_10pm to 200pm, for example from ?.10pm to 100pm; or
from _10pm to
40pm; or from ?..10pm to 30pm or any desired volume mean diameter value in
between. The
skilled addressee will appreciate that the actual VMD of particles of use in
the invention that
are used on oilseed plant seed will be appropriate to the size of the seeds to
which the
particles are to be applied. Furthermore, the skilled addressee will also
appreciate that
where the VMD is defined as being ?.10pm or ..12pm the size of the particles
will be
governed by the size of the seed to which it is applied and such a range
should be construed
as being commensurate therewith. Thus, the size range of particles of use in
the invention is
not open-ended in respect of an upper size limit but only insofar as such a
limit is applicable
to oilseed plant seed to which particles of the invention may be expected to
adhere as a
coating. The limit in the sizing of the particles of use in seed coatings of
the invention will be
apparent to the skilled addressee for oilseed plant seed.
The one or more additives for enhancing seedling vigour and/or seedling growth
from
oilseed plant seeds may be selected from one or more inorganic or chemical
additives
and/or one or more live biological agents.
Suitable inorganic agents include commercially available NPK fertilisers that
may be added
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
oilseed plant seed coatings of the invention. These may be added in the form
of dry powders
of soluble ions that include the so-called primary macronutrients such as
nitrogen,
phosphorus, and potassium; the so-called secondary macronutrients such as
calcium,
sulphur, and magnesium; and the so-called "micronutrients" (trace minerals
such as boron,
chlorine, manganese, iron, zinc, copper, molybdenum, and selenium).
"Macronutrients" are
taken up in relatively large quantities and are present in plant tissue in
quantities from about
0.2% - 4% on a dry weight basis. "Micronutrients" are taken up in smaller
quantities and are
present in plant tissue in quantities measured in parts per million (ppm),
ranging from about
5 ¨ 200 ppm, or less than 0.02% dry weight.
Additives may be selected from bioregulators commonly applied in the art such
as
brassinosteroids, cytokinines e.g. kinetin or zeatin, the auxins e.g.
indolylacetic acid or
indolylacetyl aspartate, the flavonoids and isoflavanoids e.g. formononetin or
diosmetin, the
phytoaixins e.g. glyceolline, phytoalexin-inducing oligosaccharides such as
pectin, chitin,
chitosan, polygalacturonic acid and oligogalacturonic acid, compounds such as
the
gibellerins produced by rhizobial symbionts and endophytic microorganisms such
as
acetobacter diazotrophicus and herbaspitillum seropedicae and the like.
Species of mycorrhizal fungus are also capable of augmenting levels of
available nutrients in
the soil with further organic and inorganic nutrients that are assimilable by
a crop plant.
Suitable species of mycorrhizal fungus include those that are capable of
colonising a host
plant's roots, either intracellularly as in arbuscular mycorrhizal fungi
(AMF), or extracellularly
as in ericoid mycorrhizal (EM) fungi.
Examples of AMF mycorrhizae of potential use in the invention include those
from the
Glomus, Gigaspora, Acaulospora and Sclerocystis. Suitable species include
Glomus
fasciculatum, G. intraradices, G. claroideum; G. intra, G. clarum, G.
brasilianum, G.
deserticola, G. monosporus, G. mosseae, G.tortuosum, G, sinuosum, Gigaspora
margarita,
Gigaspora gigantean and Acaulospora longular.
Ericoid mycorrhizas (EM) are known to have saprotrophic capabilities and these
are thought
to enable plants to receive nutrients from not-yet-decomposed materials via
the
decomposing actions of their ericoid partners. A suitable genus of EM of
potential use in the
invention is Pezizella.
Species of bacteria and fungi of potential use in the invention include those
that are capable
of acting on an inorganic and/or organic substrate to release phosphorus-
containing
compounds in soluble form from such substrates. Species of bacteria include
those from
Alcaligenes, Acinetobacter, Azospirillum, Bacillus, Enterobacter, Erwinia,
Flavobacterium,
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
6
Paenibacillus, Pseudomonas, Rhizobium, Burkholderia, and Serratia. Examples of
species
of the Bacillus genus are Bacillus megaterium, Bacillus coagulans, species of
the
Azospirillum genus such as Azospirillum brasilense, species of the Pseudomonas
genus,
such as Pseudomonas aeruginosa, Pseudomonas aurantiaca, Pseudomonas putida,
Pseudomonas pseudoalcaligenes, Pseudomonas fluorescens, Pseudomonas poae, and
Pseudomonas trivialis, species of the Rhizobium genus such as Bradyrhizobium
and
Rhizobium leguminosarum, and species of the Paenibacillus genus (formerly
considered as
Bacillus genus) such as Paenibacillus lautus. Commonly used Rhizobium
inoculants may be
sourced from such companies as Becker Underwood and EMD Crop Bioscience.
A further live biological inoculant that is useful for oilseed plant seed
coating is Trichoderma,
a fungus that is capable of making available, and in the adsorption of,
mineral nutrients from
the soil such as by solubilising insoluble phosphorus and zinc in the soil.
Other capabilities of
the fungus include the decomposition of organic matter thereby releasing
calcium,
potassium, and nitrogen available for plant use. By such capabilities certain
Trichoderma
species can be used to contribute to a balanced fertilisation of oilseed
plants in the field and
thereby the requirement for adding large amounts of artificial fertilisers may
be reduced by
as much as 50% depending on crop type. Trichoderma strains are known in the
art, for
example, useful strains are known from the University of the Philippines Los
Banos (UPLB),
Institute of Biological Sciences.
Examples of conventional additives for increasing fertiliser efficiency, plant
productivity,
growth, and nutrient accumulation may be sourced from such commercial sources
as
Incotec Inc., Germains, Bayer CropScience, and Becker Underwood. Suitable
additives may
be selected from commercially available products such as Auxigrow(R) (Auxein
Corp.,
Lansing, Mich., USA) and Amisorb(R) (Donlar Corp., Chicago) or the so-called
phytochelates described by A. M. Kinnersley in Plant Growth Regul. (1993),
12(3), 207-18,
which are thought to influence the availability to the plant of minimal
amounts of certain
metals such as Zn, Fe, Cu and the like for optimal growth and productivity.
Examples of the
latter include polymers of L-lactic acid, L-lactoyllactic acid and water-
soluble polyaspartates.
Other additives that may be applied to oilseed plant seed coatings of the
invention include
the kinds of adjuvant that are found in conventional commercial agrochemical
formulations.
Suitable additives for inclusion into oilseed plant seed coatings of the
invention may be
selected from those described by Chester L. Foy, Pestic. Sci.(1993) 38, pp.65-
76; and in EP
0357559. Seed coating compositions of the invention may further include
conventional
additives such as agents having wetting, dispersing and de-foaming modes of
action.
Suitable surface-active compounds are non-ionic, cationic and/or anionic
surfactants having
good emulsifying, dispersing and wetting properties. Such adjuvants for crop
protection
formulations are obtainable from fine chemicals producers [e.g. by Clariant AG
(Muttenz,
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
7
Switzerland)] and include (fatty)alcohol alkylphenol ethoxylates,
polyarylphenol ethoxylates,
dispersing phosphates, taurides and/or alcohol monosuccinates. The term
"surfactants" also
comprises mixtures of two or more surfactants and natural or synthetic
phospholipids of the
cephatin and lecithin series, e.g. phosphatidyl-ethanolamine,
phosphatidylserine,
phosphatidylglycerol, lysolecithin sugar esters. A typical de-foaming agent is
Fluowet
PL80B(R) (Clariant AG) and typical antifreeze compounds are glycols and
polyethylene
glycols. Further ingredients may include solid or liquid substances ordinarily
employed in
formulation technology, e.g. natural or regenerated minerals, tackifiers,
thickeners or
binders. Other suitable additives are emulgating protein hydrolysates, e.g. as
described in
EP 0297426 (Bayer AG). Dyes often used in seed treatment compositions include
water-
insoluble or water-soluble dyes. Examples of dyes that may be added to
compositions of the
invention include Colanyl Red(R) (Clariant AG, Muttenz), Rhodamin B, white
pigment
(titanium dioxide) or Luconyl(R) (BASF AG). Altogether additives may be used
to ensure that
the formulation disperses well, does not settle or freeze and differentiates
the seeds from
untreated seeds. Other special additives which are known to enhance seedling
vigour in
particular in combination with certain pesticides, e.g. fungicides in
combination with
3',4',5',6'-tetrachloro-2,4,5,7-tetraiodo-fluorescein (EP0297426), may be
applied to the seeds
in a combined amount that is effective, preferably synergistically effective,
to increase
seedling vigour and plant growth.
Additionally, the organic particles of use in compositions of the invention
may contain other
further components such as additives selected from UV blockers such as beta-
carotene or p-
amino benzoic acid, colouring agents such as optical brighteners and
commercially available
colouring agents such as food colouring agents, plasticisers such as glycerine
or soy oil,
antimicrobials such as potassium sorbate, nitrates, nitrites, propylene oxide
and the like,
antioxidants such as vitamin E, butylated hydroxyl anisole (BHA), butylated
hydroxytoluene
(BHT), and other antioxidants that may be present, or mixtures thereof. The
skilled
addressee will appreciate that the selection of such commonly included
additives will be
made depending on end purpose, and perceived need.
Seed compositions of the invention may be applied to oilseed plant seed at a
rate of
application from 0.1 g to 500 g, preferably from 1g to 100g, most preferably
from 5g to 50g of
the active ingredient (a.i.) per 100kg of seed.
Liquid formulations of the invention may be formulated as an aqueous
formulation or as an
oleaginous formulation, depending on design. Aqueous formulations may include
surfactants
selected from commercially available surfactants such as Tween 20, Silwet L77,
Tween 80,
Torpedo II, Newmans T80, Fortune, Guard, Rhino, Biopower, and the like.
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
8
Oleaginous formulations, that is to say oil-based formulations, may contain
any oil suitable
for use in the present invention which may be selected from petroleum oils,
such as paraffin
oil, and vegetable oils such as rapeseed oil, soybean oil, sunflower oil, palm
oil and the like.
Oil formulations of use in the invention contain organic particles of the
invention and as
described herein and these in turn may be admixed with flow agents such as
hydrophilic
precipitated silicas, for example Sipernat 383 DS, Sipernat 320, EXP 4350, and
Sipernat D-
17 and the like. Such free-flowing agents may be dispersed in oils, for
example, for anti-
foaming purposes.
The skilled addressee will appreciate that where an aqueous or an oil
formulation may be
used, the liquid element should be removed from the coated oilseed plant seeds
after
coating is achieved, for example by drying off using conventional drying
processes.
Coatings of organic materials of use in the present invention also serve to
protect
immediately planted oilseed plant seeds from soil borne pathogens, that is to
say, pathogens
that are able to colonise the seed cuticle and/or pathogens that populate the
soil and which
are capable of acting on oilseed plant seeds. Such soil borne pathogens are
typically
bacteria and/or fungi. Examples of soil borne bacterial and fungal pathogens
that attack
oilseed plants include Rhizoctonia spp. (active against e.g. rapeseed, canola;
R. solani
active against B. napus and B. campestris), Peronospora spp. such as P.
parasitica (active
against B. napus and B. campestris) Pythium spp, (active against e.g. canola
and oilseed
rape, sunflower), Fusarium spp. (active against sunflower) such as F.
oxysporum (active
against e.g. canola and oilseed rape), Phytophthora spp. (active against e.g.
canola and
oilseed rape e.g. P. megasperma), Vedic'Ilium spp. (active against sunflower)
such as V.
Iongisporum (active against Brassica spp. such as B. napus and B. campestris),
Sclerotium
spp. (active against e.g. canola), Agrobacterium tumefaciens (active against
sunflower and
Brassica spp. e.g. canola), Phoma spp.(active against sunflower) such as Phoma
lingam
(active against Brassica spp.), Pseudomonas spp. (active against sunflower;
and canola e.g.
P. syringae pv maculicola), Altemaria spp. (active against canola, sunflower),
and the like.
According to a further aspect of the invention there is provided use of an
organic material in
the form of particles wherein the organic material is selected from at least
one wax having a
melting point of ?.50 Centigrade in the manufacture of a coating composition
for oil seeds.
The organic materials are selected from one or more organic materials having a
melting
point of .50 Centigrade, more preferably of ?..60 C and most preferably are
made up of hard
waxes having a melting point of .70 C. Suitable organic materials include
those as
described herein, such as carnauba wax, beeswax, montan wax, Chinese wax,
shellac wax,
spermaceti wax, candelilla wax, castor wax, ouricury wax, and rice bran wax or
a mixture of
two or more thereof, and preferably, the seed coating that is used includes
carnauba wax.
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
9
Preferably, in this aspect of the invention, the organic particles have a mean
volume
diameter selected from ?.10pm to 200pm, as herein described and as appropriate
for the
oilseed plant seeds to which the particles are to be applied. Naturally, the
skilled addressee
will appreciate that the size of the organic particles to be applied to
oilseed plant seeds will
depend on the size of the seed, and the type or form of such oilseed plant
seeds that are
contemplated for coating.
In a third aspect of the invention there is provided a method of manufacturing
an oilseed
coating composition as herein described that comprises
1) selecting solid organic material wherein the solid organic material is wax
having a melting
point of _50 C;
2) machining said organic material into particles of a volume mean diameter of
?_10pm; and
3) adding one or more additives for enhancing seedling vigour and/or seedling
growth
selected from one or more inorganic additives and/or one or more live
biological agents.
The organic material in this aspect of the invention may be selected from
organic materials
such as from organic waxes having a melting point of ?_50 C, more preferably
of ?..60 C, and
most preferably are made up of hard waxes having a melting point of _70 C.
Suitable waxes
for use in this aspect of the invention include carnauba wax, beeswax, montan
wax, Chinese
wax, shellac wax, spermaceti wax, candelilla wax, castor wax, ouricury wax,
and rice bran
wax or a mixture of two or more thereof. Preferably, the selected organic
material includes a
substantial proportion of carnauba wax up to 100%, for example 1%, 10%, 20%,
30%, 40%,
50%, 60%, 70%, 80%, 90% or more or any proportion therein between, the rest
being made
up of is at least one other organic material as herein defined. Preferably,
the selected
organic material is solely carnauba wax which may contain further additives as
herein
described, and further components such as UV blockers, antioxidants such as
vitamin E and
the like.
In a further aspect of the invention, there is provided an oilseed plant seed
coating
composition produced by the method as described herein.
In a further aspect of the invention there is provided a coating composition
as described
herein for use on oilseed plant seeds.
In a further aspect of the invention there is provided a method of coating
oilseeds with a
coating composition that comprises at least one organic material as herein
defined wherein
the organic material is selected from waxes having a melting point of 50
Centigrade, the
method comprising
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
i) obtaining organic material as a population of particles of a pre-determined
VMD; and
ii) applying the said population of particles to oilseeds.
The skilled addressee will appreciate that environmental factors that may
affect oilseed plant
seed viability includes such factors as extremes of heat, loss of moisture and
the presence
of pathogens such as bacteria and/or fungi. The skilled addressee will also
appreciate that
the pre-determined VMD will be appropriate to the size of the oilseed plant
seeds, to which
the coating is to be applied.
In a variant of this aspect of the invention there is provided a method of
coating oilseeds with
a coating composition that comprises an organic material wherein the organic
material is
selected from waxes having a melting point of 50 Centigrade, the method
comprising
i) obtaining said material;
ii) heating the organic material so as to form a liquid phase or a gaseous
phase;
iii) cooling the liquid phase or gaseous phase of ii) to below the melting
point of the
organic material, forming a solid;
iv) adding one or more additives to the solid formed in iii);
v) machining the solid organic material of iii) into particles of a pre-
determined VMD;
and
vi) applying the particles of v) to oilseeds.
In a second variant of the above aspect of the invention there is provided a
method of
coating oilseeds with a coating composition of the invention that comprises an
organic
material wherein the organic material is selected from waxes having a melting
point of
.50 Centigrade, the method comprising
i) obtaining said organic material;
ii) heating the organic material of i) so as to form a liquid phase or a
gaseous phase;
iii) adding one or more additives to the liquid phase or gaseous phase of
ii);
iv) cooling the liquid phase or gaseous phase of iii) to below the melting
point of the
organic material, forming a solid;
v) machining the solid organic material of iv) into particles of a pre-
determined VMD as
herein defined; and
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
11
vi) applying the particles of v) to oilseeds.
The oilseeds are typically selected from oilseed plants as herein defined. The
organic
material of use in the invention may comprise one or more organic materials
selected from
organic materials as herein defined. Preferably, the organic material is
carnauba wax. Where
two or more organic materials of use in the invention are employed as the
organic material in
for example, an oilseed coating composition of the invention they may be
heated together so
as to form a liquid phase or a gaseous phase during which phases the organic
material may
be mixed, if required. Once the organic materials are mixed they may be cooled
to below the
melting point of the organic material possessing the lowest melting point in
the liquid phase
(where a gas phase is employed, this will be cooled to a liquid phase),
forming a solid which
may then be machined, such as by comminution, into particles of a pre-
determined VMD as
herein defined using conventional procedures. As described above, one or more
additives
may be added to the organic materials at points indicated above. It will be
appreciated that
the person skilled in the art will understand at what point or points in the
described
processes additives may be added to the organic material, depending on the
additive
material to be added to the organic material forming particles of use in the
invention.
Once the organic material is in the form of particles of a known VMD, the
particles may be
applied to oilseed plant seeds using conventional means, such as by tumbling.
The treatment composition is applied to oilseed plant seeds, in dry
particulate form or liquid
form as hereinbefore described, and preferably in dry particulate form. The
organic material
in the above aspect and variant aspect of the invention may be selected from
organic
materials selected from organic waxes having a melting point of ?_50 C, more
preferably of
?_60 C, and most preferably are made up of hard waxes having a melting point
of _70 C.
Suitable waxes for use in the invention include those waxes as herein before
described and
include carnauba wax, beeswax, montan wax, Chinese wax, shellac wax,
spermaceti wax,
candelilla wax, castor wax, ouricury wax, and rice bran wax or a mixture of
two or more
thereof. Preferably, the selected organic material includes a substantial
proportion of
carnauba wax up to 100%, for example 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%,
80%,
90% or more or any proportion therein between, the rest being made up of at
least one other
organic material as herein defined. Preferably, the selected organic material
is solely
carnauba wax which may contain further added components as herein defined,
such as UV
blockers, antioxidants such as vitamin E and the like.
Generally, the particles of use in the above aspect of the invention and the
variant aspect of
the invention possess a volume mean diameter as herein before described.
There now follow examples that illustrate the invention. It is to be
understood that the
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
12
examples are not to be construed as limiting the invention in any way.
Example: Growth and Vigour in B. napus.
Brassica napus seed provided by Herbiseeds (Twyford, UK)
Combination of Carnauba wax particles and Inoculant
Rock Phosphate
Rock Phosphate (Garden Direct,UK) with a 30% P205 content is crushed using a
pestle and
mortar and then passed through a 32 micron mesh sieve.
Carnauba Wax Sizing Method
Steps in Air Milling in Boyes Micronisation Process (for carnauba wax
particles with a VMD
of approx. 75pm)
,
1. 2kg camauba wax blocks are first kibbled into approximately 4 to 6mm pieces
in a KT
Handling Ltd Model 04 kibbler (serial no. 729/C) following the manufacturer's
instructions.
2. The kibbled pieces are then passed through an Apex Construction Ltd Model
314.2
Comminuting Mill (serial no. A21306) and reduced further in size to a range of
250 to
300um.
3. The comminuted particles are then passed through a Hosokawa Micron Ltd
Alpine
100AFG jet mill (serial no. 168092) following the manufacturer's instructions,
setting the mill
at a speed of 2,500rpm with a positive system pressure of 0.03bar.
4. The grinding air is to be kept to 6 bar, the system rinsing air flow and
Classifying Wheel
gap rinsing air are both to be set at a minimum of 0.5 bar and no more than
0.75bar, the
cleaning air filter is to register a delta of no more than 5bar to achieve a
final particle size
with a VMD of approx. 75um.
Rock phosphate is combined with Carnauba wax particles (VMD 75pm) at a ratio
of 1:3
(Rock Phosphate:Carnauba wax particles). A homogeneous mix is attained through
tumbling
seed and camauba wax formulation in a cylinder, adapted to produce lateral
mixing/tumbling
through the inclusion of angled interior vanes, placed on a Wheaton roller for
5 minutes.
Dry Powder Formulation of Mycorrhizae International Culture Collection of VA
Mycorrhizal Fungi (INVAM).
Mycorrhizae concentration is measured by diluting 1gm in 11 of water, before
further diluting
CA 02841043 2014-01-06
WO 2012/143684 PCT/GB2012/000366
13
by taking 1 ml of the suspension and making it up to 1000m1. A 20p1 sample is
then added to
an Improved Neubauer Counting Slide and a count made of 4 large squares
(0.1mrnA3) in
both of the grids. The mean for each square is calculated and the mean of the
two grids
used to produce a measurement of spores per 100n1 of water. The dilution
factor is then
applied to produce an approximation of the number of spores per gram.
Carnauba wax particles are produced following the above descrobed procedure
with the
exception that the milling speed was set at 8000rpm to obtain wax particles
having a VMD of
of 16pm. The particles are combined with Mycorrhizae at a ratio of 1:3
(Mycorrhizae:Carnauba wax particles) in a 50mItube using a Stuart roller mixer
set at 25rpm
for 5 minutes. This can then be used to calculate the quantity of
spore/Carnauba wax
particles powder mix required for the seed coating based on a standard of
1x104 spores
gram-1 of seed.
A homogeneous mix of is attained through tumbling seed and carnauba wax
formulation in a
cylinder, adapted to produce lateral mixing/tumbling through the inclusion of
angled interior
vanes, placed on a Wheaton roller for 5 minutes.
=
I Chitosan
Chitosan (>75% Deacetylated chitin, Poly(D-glucosamine)) (Sigma Aldrich,UK) is
crushed
using a pestle and mortar and then passed through a 32 micron mesh sieve.
Chitosan is combined with Carnauba wax particles obtained using the protocol
described
above with the exception that the milling speed was set at 2,500 rpm to obtain
particles
having a VMD of approx. 75pm at a ratio of 1:19 (Chitosan:Carnauba wax
particles). A
homogeneous mix of is attained through tumbling seed and carnauba wax
formulation in a
cylinder, adapted to produce lateral mixing/tumbling through the inclusion of
angled interior
vanes, placed on a Wheaton roller for 5 minutes.
Treatments:
1. Carnauba wax particles and Mycorrhizae
2. Carnauba wax particles and Rock Phosphate
3. Carnauba wax particles and Chitosan
4. Mycorrhizae control
5. Rock Phosphate control
6. Chitosan control
7. Carnauba wax particle control (vehicle control)
8. Untreated Control
Seeds are planted in two 84 well plug trays using moist seed potting compost
(John Innes
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
14
No.2). The trays are placed in a Vitopod propagator (Greenhouse Sensations,
UK) at 20 C.
Moisture content (Brannan Soil Moisture Meter, Fisher Scientific, UK) and pH
levels
(Brannan Soil pH meter, Fisher Scientific, UK) are checked to ensure that the
conditions are
consistent across the tray. The order of the treatments is randomised (by row
units) to
reduce any unforeseen bias.
At the true leaf stage the plants are carefully transplanted from the plugs to
7cm square pots
filled with a sterilised top soil. The macro-nutrient (nitrates, phosphates
and potassium)
content of the top soil is measured using a La Motte Model STH-4 soil testing
kit and
recorded. Six replicates for each treatment (48 plants) are randomly assigned
to each of
three propagators, and further randomised within the propagator (total = 144
plants). The
propagators are set at 15 C, 20 C and 25 C. Light is provided on a 16:8
Light:Dark cycle
using a twin bulb T5 lighting array suspended 150mm above the propagator
(Lightwave T5,
48w, 3300 lumens). T5 tubes (6500 Kelvin) deliver the bright blue/white light
required by the
plant for growth without emitting much heat which may scorch tender seedlings
Moisture content and pH levels are checked to ensure that the conditions are
consistent
across the propagator by measuring six random plants along a pathway
(alternating between
a W and Z). This is repeated for each propagator.
Plants are watered as required based on conditions to maintain consistent soil
moisture
content of 18% throughout all plants.
The lids of the propagators are removed at such time as required due to the
plant height.
After 21 days the plants are removed from the propagators and the following
measurements
recorded:
Root weight (fresh)
Shoot weight (fresh)
% Mycorrhyzal root colonisation (by microscopic examination)
Plant tissue is measured for macro-nutrient content using the instructions
provided with a La
Motte Model PT-3R Plant Tissue Test kit.
Analysis
The percentage data (root colonisation data) were arcsine transformed. The
influence of the
factors and their interactions are tested with a two-way analysis of variance.
Where the
ANOVA reveals significant effects by the factors, the differences between
treatments are
separated using a post hoc least significant difference (LSD), multiple
comparison test (p 5
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
0.05).
The influence of the factors and their interactions are tested with a 2-way
ANOVA. The
analysis was done for each temperature separately and with temperature as a
factor. For the
ANOVA with temperature as a factor, treatments were used as a sub-plot factor.
Fisher's
Least Significance Differences were calculated at the 5% significance level to
compare
treatment means. Shapiro-Wilks's test was performed to test for non-normality
The above procedures are followed to apply: rock phosphate, chitosan and
Glomus sp. on
seeds of canola (Brassica campestris).
The above procedures are followed to apply: rock phosphate, chitosan and
Glomus sp. on
seeds of Sunflower (Helianthus annuus).
Delivery of Macronutrients using Carnauba wax particles as a seed coating on
Oilseed
Rape
IAim: to assess the potential for formulating essential macronutrients into
carnauba wax
particles and using this as a seed coating to provide the germinating seed and
seedling with
supplementary nutrients to aid in early stage growth.
Macronutrients selected:
Phosphorus (P)
= Phosphorus (P) is an essential part of the process of photosynthesis.
= Involved in the formation of all oils, sugars, starches, etc.
= Helps with the transformation of solar energy into chemical energy;
proper plant
maturation; withstanding stress.
= Effects rapid growth.
= Encourages blooming and root growth.
Potassium (K)
= Potassium is absorbed by plants in larger amounts than any other mineral
element
except nitrogen and, in some cases, calcium.
= Helps in the building of protein, photosynthesis, fruit quality and
reduction of
diseases.
= Potassium is supplied to plants by soil minerals, organic materials, and
fertilizer.
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
16
Both Potassium and Phosphorus can be found in soluble form in Monobasic
Potassium
Phosphate or MKP(KH2PO4), a soluble salt commonly used as a fertiliser and
plant growth
supplement.
Formulation Method
Carnauba is heated on a hotplate at 100 C to a molten state. Monopotassium
phosphate
(MKP) is dissolved in deionised water to the required concentration. The MKP
solution is
slowly added to the molten wax under stirring at 1500rpm. Stirring continues
for 5 minutes
before the water/wax emulsion is poured onto a metal sheet to cool. The
resulting solid wax
including micro-droplets of MKP is then micronized in an air mill following
the procedure
described above with the exception that the milling speed was set at 12,500rpm
in order to
obtain particles having a VMD or approx. 10.3pm.
Experimental Method
Wax particles containing 10% MKP are added to 10g of oilseed rape seed, cv.
Sesame (LS
Plant Breeding, UK), at loadings of 0.1% and 1% by mass. Seed is well mixed to
ensure a
homogenous distribution across the seed. A third batch of seed is combined
with
unformulated carnauba wax particles as a control.
seeds for each treatment are sown in 20 cell modular seed trays with an
individual cell
size: Length 37mm x Width 37mm x Depth 65mm, with each tray representing a
single
sample. Each treatment is replicated four times.
The pots are filled with a sieved, heat-sterilised seed mix (Levingtons F1
Seed and Modular
Compost ¨ Low Nutrient)) to level with the top of the cell. Low Conductivity:
250-280 pS,
Standard pH: 5.3-5.7, Mg/litre added: N ¨ 100, P ¨ 200, K ¨ 200.
They are then lightly tamped and 30m1 of deionised water added to each cell
through a
course filter. A single seed is then placed on the surface of the soil and
covered with a thin
layer of vermiculite to a depth of 2-3x the diameter of the seed, as per
supplier
recommendation. The trays are placed within plastic gravel trays (two per
tray) which are
lined with capillary matting to aid watering.
The gravel trays are then placed in a thermostatically controlled plant growth
chamber
(Fitotron SGC120, Weiss Gallenkamp, Loughborough, UK). Temperature cycling is
set at
C/10 C on a 16/8hr schedule. Lighting at 150 pmol m-2 s-1 on a 16/8hr
photoperiod is
introduced at first emergence.
Plants are watered daily from the bottom in order to maintain a compost
moisture level of
approximately 40% in the cells
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
17
After 10 days the plants are removed from the individual cells and the compost
mix
separated from the root structure. Plants from each 10 cell tray are combined
and separated
into shoots, made up of the first true leaves and growing tip, and roots.
PLANT TISSUE ANALYSIS: (conducted to the following method by NRM Laboratories
(Bracknell, UK)
Total Phosphorus (P), Potassium (K), Magnesium (Mg), Calcium (Ca), Sodium
(Na),
Manganese (Mn), Copper (Cu), Iron (Fe), Zinc (Zn), Boron (B) determination
using
Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) method
EQUIPMENT
1. ICP Emission Spectrograph
2. Autosampler
3. Digital Dilutor
REAGENTS
1. 3N Hydrochloric Acid: Dilute 250 ml. of concentrated hydrochloric acid to 1
litre using
deionized water and mix well.
2. Nitric Acid: HNO3
STANDARDS
1. Stock Solutions: Use 1000 PPM certified, NIST traceable, plasma grade
standards
for the 16 listed elements.
2. Instrument Calibration Standards:
a. WAT 1 - deionized water
b. WAT 2 - Mn, Fe, Al, B, Cu, Zn, Na, Pb, Cd, Ni, Cr, Mo - Pipet 10 ml. of
stock
solution of each element into a 1 litre volumetric flask. Add 5 ml. of nitric
acid.
Dilute to volume with deionized water and mix well.
c. PLN2 - P, K, Ca, Mg - Pipet the designated ml. of stock solution into a 1
litre
volumetric flask. Add 5 ml. of nitric acid. Dilute to volume with deionized
water
and mix well.
CA 02841043 2014-01-06
WO 2012/143684 PCT/GB2012/000366
18
Final
Stock Solution Concentration Instrument
Element ml. forEJ:_-). Readout %
_
P 10 10 1.00
K 50 50 5.00
Ca 20 20 2.00
Mg 10 10 1.00
3. Instrument Calibration Verification Standards:
a. A second set of calibration standards obtained from a different
manufacturer.
SAMPLE PREPARATION
Samples are dried and ground to pass through a 1mm screen.
The elements in the residue remaining after the destruction of the organic
matter by ashing
at 550 C are dissolved in hydrochloric acid:
1. Dry Ash
a. Weigh 1g sample into a 10 ml. glazed, high-form porcelain crucible.
b. Ash in a muffle furnace for 4 hours at 500 C.
c. Let cool and add 5 ml. of 3N HCI.
d. Place on a hot plate and boil gently for 5 minutes.
e. Let cool and transfer to a 100 ml. volumetric flask. Dilute to volume with
deionized water and mix well. Use this solution for the analysis of Mn, Fe,
Al,
B, Cu, Zn, Na, Pb, Cd, Ni, Cr and Mo.
f. Dilute the solution obtained in le. one to ten with deionized water using a
digital dilutor. Use this solution for the analysis of P, K, Ca and Mg.
ICP PROCEDURE
1. Set up and operate the ICP Emission Spectrograph in accordance with
manufacturer's specifications.
2. Mn, Fe, B, Cu, Zn, Na, Ni analysis.
a. Choose PLANT from the method menu.
b. Calibrate the instrument using WAT1 and WAT2 instrument calibration
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
19
standards.
c. Analyze the sample digests obtained in le. of the sample preparation
section.
3. P, K, Ca, Mg analysis.
a. Choose PLANTDIL from the method menu.
b. Calibrate the instrument using WAT1 and PLN2 instrument calibration
standards.
c. Analyze the digests obtained in 1f. of the sample preparation section.
QUALITY CONTROL
1. Following calibration, analyze one high instrument calibration standard,
one
instrument calibration verification standard and one quality control sample.
a. Instrument Calibration Standard: Values must be within 3% of the known
value for K and Mo. All other elements must be within 2% of the known value.
b. Instrument Calibration Verification Standard: Values must be within 10% of
the certified values.
c. Quality Control Sample: Values for all elements must be within limits
established by the Extension chemist.
2. Analyze a high instrument calibration standard after each tenth sample and
at the
end of the set of samples.
a. Values must be within 8% of the known values.
b. If any of the values are greater than 8% from the known values, recalibrate
the instrument and begin sample analysis from the last "good" instrument
calibration standard.
3. Prepare one duplicate sample for each 10 samples. If the set contains less
than 10
samples, prepare one duplicate per set.
a. Results on the duplicate sample should agree within 20% of the average
value of the two samples.
REFERENCES
1. Isaac, R.A. and W.C. Johnson, 1985, Elemental Analysis of Plant Tissue by
Plasma
Emission Spectroscopy: Collaborative Study. JAOAC. 68(3), pp 499- 505.
2. AOAC Official Method 985.01, in Official Methods of Analysis of AOAC
International, 16th
edition, Volume I Chapter 3, p. 4.
3. AOAC Official Method 968.08 D(a), in Official Methods of Analysis of AOAC
International,
16th edition, Volume I Chapter 4, p. 23.
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
Phosphate Solubilisation using Beneficial Microbes
Several bacterial species are able to impart a beneficial effect upon plant
growth. Mostly
they are associated with the plant rhizosphere, so they are called as
rhizobacteria. This
group of bacteria has been termed plant growth promoting rhizobacteria, and
among them
are strains from genera such as Alcaligenes, Acinetobacter, Arthrobacter,
Azospirillum,
Bacillus, Burkholderia, Enterobacter, Erwinia, Flavobacterium, Paenibacillus,
Pseudomonas,
Rhizobium, and Serratia.
The production of organic acids by phosphate solubilizing bacteria has been
well
documented and identified as the main mechanism for phosphate solubilisation.
Gluconic
acid seems to be the most frequent agent of phosphate solubilisation
(Pseudomonas sp.),
and 2-ketogluconic acid is also identified in strains with phosphate
solubilizing ability
(Rhizobium sp.).
Saprophytic fungi are also known to solubilise both organic and inorganic
phosphates.
Several genus, including Trichoderma, Penicillium, and Gliocladium have
exhibited potential
as biofertilisers. Morales et al (2011) demonstrated that Penicillium albidum
was able to
solubilise 64mg of organic/inorganic phosphate per gram of fungi.
Experiment to assess the potential for delivery of Phosphate Solubilising
Organisms
as a seed costing using Carnauba wax particles
Using a dry spore powder of a phosphate solubilising organism, such as
Penicillium bilaii.
Spores are combined with carnauba wax particles with a VMD of approximately
10pm at a
ratio of 1:3. The powders are agitated to create a homogenous mix and applied
to sterilised
Oilseed rape seed at a loading of 0.1% (by mass). Additional batches of seed
are treated
with spores only (0.1%), Entostat only (0.1%) and untreated seed.
Phosphate Solubilising Activity Screening
Plate screening using Pikovskays' medium (see below) is used to demonstrate
phosphate
solubilising activity of the treated seed. 9cm petri dishes are divided into
quadrants and a
seed is placed in the centre of each quadrant. Plates are incubated at 20 C
for 4 days.
Active phosphate solubilising agents produce clear zones around the seed as
they solubilise
the insoluble mineral phosphates within the media. The radius of the clear
zones is
measured and compared to the mean results achieved for each treatment.
Differences are
CA 02841043 2014-01-06
WO 2012/143684
PCT/GB2012/000366
21
analysed using one-way ANOVA and Tukey Post-Hoc diagnostic test where
significance is
found.
Phosphate uptake by plant
Seeds are treated as described above.
Sure to Grow PET grow cubes (25x25x38mm) are soaked in deionised water
containing 1%
Ca3(PO4)2 in suspension until saturated. Cubes are placed in free draining
plant trays on a
level surface to prevent nutrient run-off and migration whilst taking care to
avoid pooling of
water at the root zone. 10 cubes are used per tray and the mean of these
represents one
replicate. Each treatment is replicated 8 times.
A single Oilseed rape seed is placed in the cross-cut X in the top of each
cube. Seed trays
are then covered to maintain a humid environment and regularly top watered
with the 1%
Ca3(PO4)2 suspension to maintain a moist cube. Trays are incubated at 20 C and
10 C on a
16/8hr thermal cycle. On germination the cover is removed and the seedling
exposed to
lighting on a 16/8hr photoperiod.
After 15 days the plants are removed from the grow cube and nutrient content
of the plant
tissue is analysed using the ICP method described above.
Differences in the Phosphate content between treatments are assessed
statistically using
one-way ANOVA.
Pikovskays' Medium
Components Quantities (g r)
Glucose 10
Ca3(PO4)2 5
(NH4)2SO4 0.5
NaCI 0.2
MgSO4.7H20 0.1
KCI 0.2
Yeast Extract 0.5
MnSO4.H20 0.002
FeSO4.7H20 0.002
pH 7.0
