Note: Descriptions are shown in the official language in which they were submitted.
2 1 99368
WO 96/07319 PCTllil95100484
, Improving the yield of plants
, Technical field
The invention relates to the use of betaine to
improve the yield of plants. The invention relates
especially to the use of betaine to improve the yield
of plants belonging to the genus Sol anum . According to
the invention, the yield can be improved particularly
under stress conditions, i.e. when the conditions are
poor due to e.g. low temperatures, drought, high
salinity or env ro~mental poisons interfering with the
growth. The invention also relates to plants of the
genus Sol anum treated with betaine and to products
obtained from these plants.
Background
The environment and conditions of growth
considerably affect the growth result of plants. Optimum
growth environment and conditions usually result in a
yield that is large in quantity and high in quality.
Under poor growth conditions both the quality and the
quantity naturally deteriorate.
Several different solutions have been developed
to improve the growth conditions and yield of plants.
Selecting the right plant for the right growth location
is self-evident for a person skilled in the art. During
the growing season plants may be protected with
mechanical means by utilizing for example different
gauzes or plastics or by cultivating the plants in
greenhouses. Irrigation and fertilizers are generally
used in order to improve the growth. The physiological
properties of a plant are manipulated by means of
breeding, both with traditional breeding methods and for
example with genetic manipulation. The methods are often
laborious and impractical, their effect is limited (the
I ~ 2199368
WO96/07319 PCT~S/00484
economical size of a greenhouse, the limited protection
provided by gauzes, etc.), and they are also far too
expensive on a global scale. No economically acceptable
chemical solutions for protecting plants from
environmental stresses have been described so far.
Plants can adapt to some extent to stress
conditions. Under these circumstances for example
proline and betaine are accumulated in the regions of
growth of certain plants. The literature of the art
discusses the function and meaning of these accumulated
products. On the one hand it has been proposed that the
products are by-products of stress and thus harmful to
the cells, on the other hand it has been estimated that
they may protect the cells (Wyn Jones, R.G. and Storey,
R.: The Physiology ~nd Biochemistry of Drought
Resistance in Plants, Paleg, L.G. and Aspinall, D.
(Eds.), Academic Press, Sydney, Australia, 1981).
Zhao et al. (in J. Plant Physiol. 140 (1992)
541 - 543) describe the effect of betaine on the cell
membranes of alfalfa. Alfalfa seedlings were sprayed
with 0. 2M glycinebetaine, whereafter the seedlings were
uprooted from the substrate, washed free of soil and
exposed to temperatures from -10~C to -2~C for one hour.
The seedlings were then thawed and planted in moist sand
for one week at which time regrowth was apparent on
those plants that had survived. Glycinebetaine clearly
improved the cold stability of alfalfa. The effect was
particularly apparent at -6~C for the cold treatment.
All controls held at -6~C for one hour died, whereas 67%
of the seedlings treated with glycinebetaine survived.
Itai and Paleg (in Plant Science Letters 25
(1982) 329 - 335) describe the effect of proline and
betaine on the recovery of water-stressed barley and
cucumber. The plants were grown in washed sand, and
polyethylene glycol (PEG, 4000 mol. wt.) was added to
~ WO96/07319 2 1 9 9 3 6 8 PCT~95~00~84
the nutrient solution for four days in order to produce
water stress, whereafter the plants were allowed to
recover for four days before harvesting. Proline and/or
betaine (25 mM, pH 6.2) was sprayed on the leaves of the
plant either on the first or third day of the stress or
;mm~ jately before harvesting. As regards barley, it was
noted that betaine supplied either before or after the
stress had no effect, whereas betaine added in the end
of the stress was effective. Proline had no effect. No
positive effect was apparent for cucumber. On the
contrary, it was found out that both betaine and proline
had a negative effect.
Brief description of the invention
In connection with the present invention it has
now surprisingly been found that the yield of plants,
such as potato and tomato, belonging to the genus
Sol anum can be considerably improved by means of
exogenously applied betaine. Betaine is especially
effective when the plants are subjected to exogenous
stress during the growth.
The invention thus relates to the exogenous use
of betaine to improve the yield of plants belonging to
the genus Sol anum . The invention relates especially to
the use of betaine to improve the yield of plants under
stress conditions.
The invention also relates to plants of the
genus Solanum treated exogenously with betaine, to the
products of the plants, and to their use as such or as
raw material for food industry.
The invention also relates to a method of
improving the yield of plants of the genus Sol anum, in
which method growing plants of the genus Sol anum are
exogenously treated with betaine.
~' .r -~ .'i 21 99368
WO96/07319 PCT~5/00484
The invention further relates to plants of the
genus Sol anum obtained with the method according to the
invention and to the products of these plants.
Betaine is applied to the plant in either one
or several successive treatments. The application may
be performed for example by spraying together with some
other spraying of fertilizers or pesticides, if desired.
Betaine utilized according to the invention is
transported to the plant cells, actively regulates the
osmotic balance of the cells and also participates in
other processes of cell metabolism. A cell treated with
betaine is more vizbl~ ~ven wher. subjected to exogenous
stress factors.
The betaine treatment according to the
invention is economically advantageous, and the yield
increases in an amount that is economically profitable
and significant. The treatment does not produce
significantly more work since it may be performed
together with other sprayings of fertilizers or
pesticides, and it does not require new investments in
machinery, equipment or space. It should also be noted
that betaine is a non-toxic natural product, which has
no detrimental effects on the quality of the yield.
Betaine is also a stable substance that remains in the
plant cells and thereby has a long-standing effect.
Detailed description of the invention
Betaine refers to fully N-methylated amino
acids. Betaines are natural products that have an
important function in the metabolism of both plants and
animals. One of the most common betaines is a glycine
derivative wherein three methyl groups are attached to
the nitrogen atom of the glycine molecule. This betaine
compound is usually called betaine, glycinebetaine or
trimethylglycine, and its structural formula is
presented below:
~ WO 96/0731~ 2 1 9 9 3 6 8 PCT/F~95/00484
CH3
., I
CH3 - N ' - CH2C00-
.' I
CH3
Other betaines are for example alaninebetaine
and prolinebetaine, which has been reported to for
example prevent perosis in chicks. R.G. Wyn Jones and
R. Storey describe betaines in detail in The Physiology
and Bioche~istry of Drought Resistance in Plants (Paleg,
L.G. znd Asp~nall, D. (Eds.), Academic Press, Sydney,
Australia,-1981).
Betaine has a bipolar structure and it contains
several chemically reactive methyl groups which it can
donate in enzyme-catalyzed reactions. Most organisms can
synthesize small amounts of betaine for example for the
methyl function, but they cannot react to stress by
substantially increasing the production and storage of
betaine. Best known organisms accumulating betaine are
plants belonging to the Chenopodiaceae family, for
example sugar beet, and some microbes and marine
invertebrates. The main reason for the betaine
accumulation in these organisms is probably that betaine
acts as an osmolyte and thus protects the cells from the
effects of osmotic stress. One of the main functions of
betaine in these plants and microbes is to increase the
osmotic strength of the cells when the conditions
require this, for example in case of high salinity or
drought, thus preventing water loss. Unlike many salts,
betaine is highly compatible with enzymes, and the
betaine content in cells and cell organelles may
therefore be high without having any detrimental effect
on the metabolism. Betaine has also been found to have
a stabilizing effect on the operation of macromolecules;
WO96/07319 2 1 9 9 3 6 8 PCT~5/00484 ~
it improves the heat resistance and ionic tolerance of
enzymes and cell membranes. Plants of the genus Solanum
do not normally store betaine in their cells.
Betaine can be recovered for example from sugar
beet with chromatographic methods. Betaine is
commercially available for example under the trademark
of BETAFIN, Cultor Oy, Finnsugar Bioproducts. BETAFIN
is crystalline water-free betaine of Finnsugar
Bioproducts. Other betaine products, such as betaine
monohydrate, betaine hydrochloride and raw betaine
liquids, are also commercially available and they can
be used for the purposes of the present invcntisn.
According to the present invention, betaine is
used exogenously in order to improve the yield of plants
belonging to the genus Solanum. Betaine is used
especially when plants are grown under stress
conditions, i.e. when the plants are subjected to
periodic or continuous exogenous stress. Such exogenous
stress factors include for example drought, humidity,
low or high temperatures, high salinity, herbicides,
environmental poisons, etc. Treating plants subjected
to stress conditions exogenously with betaine for
example improves the adaptation of the plants to the
conditions and maintains their growth potential longer,
thereby improving the yield-producing capacity of the
plants. Betaine is also a stable substance that remains
in the plant cells. The positive effect of betaine is
thereby long-standing and diminishes only gradually due
to dilution caused by the growth.
Betaine is applied to plants in either one or
several successive treatments. The amount used varies
depending on the plant variety and the phase of growth.
For example in the case of potato, about O.l to 20 kg
of betaine can be used per hectare. A useful amount is
thus for example about l0 kg of betaine per hectare,
. . - ' 21 99368
WO96107319 PCT~S/00484
which corresponds to about 0.01% of the potato biomass.
. A preferable amount is about 2 to 8 kg of betaine per
hectare. For tomato, about O.l to 30 kg of betaine per
, hectare can be used. A preferable amount is about l to
6 kg/ha, particularly about 2.5 kg/ha. The amounts given
here are only suggestive; the scope of the present
invention thus contains all amounts that work in the
manner described herein.
Any method suitable for the purpose can be
utilized for the application of betaine. Betaine can
easily be spread for example through spraying. Such
spraying can be perfcrm2d tcgether with some other
spraying of fertilizers or pesticides, if desired.
According to the invention, an aqueous solution of
betaine is preferably used.
The time of the treatment according to the
invention may vary, and a suitable time is determined
preferably separately for each plant. If betaine is
applied in a single treatment, the treatment is usually
performed at an early stage of growth, for example on
plants of about 5 to 20 cm. If betaine is applied in two
successive treatments, the second spraying is performed
preferably in the beginning of flowering or when stress
can be forecasted on the basis of the weather.
The betaine treatment according to the
invention considerably improves the yield of plants
belonging to the genus Solanum, for example the amount
and quality of the yield. The treatment according to the
invention is economically advantageous and the increase
in the yield is economically profitable and significant.
t For example the amount of a potato yield has been
increased by more than 30~, and for tomato the amount
of yield has been as much as doubled with a suitable
application rate of betaine. It must also be noted that
a cell treated with betaine or proline remains viable
- ~ 21 q9368
WO 96/07319 PCT/FI9~/00184
even when subjected to exogenous stress factors, such
as low temperatures, drought, high salinity, or the
like.
The invention will be described in greater
detail by means of the following examples. These
examples are only provided to illustrate the invention,
and they should not be considered to limit the scope of
the invention in any way.
Example 1
The effect of betaine on potato yield was
determined under field conditions in two different
locations and utilizing four different betaine
concentrations: 0 (control), 1.25, 5.0 and 10 kg of
betaine per hectare. For the purpose of dosage, an
aqueous solution of betaine was prepared, the solution
containing 2 ml/l of non-ionic wetter, Plus-50 (Ciba
Geigy), in addition to the desired betaine content.
Betaine solution was added in an amount of 640 l/ha at
75% ground cover, and a second application was made
during the tuber growing stage. The potato cultivar was
Russet Burbank. The places of growth varied for climate,
in one (1) the climate was warmer and drier than in the
other (2) where frost occurred during the growing
season. After the harvesting the tubers were graded into
unmarketable (small, green and odd-shaped tubers) and
marketable ones, and the weight and number of tubers in
the categories were determined. The specific gravity of
the tubers was determined with the weight in air-weight
in water method. Statistical analysis of the results was
performed by means of variance analysis utilizing
Genstat statistical package.
In location (l), tuber yield per plant
increased from a control value 1.96 kg to 2.42 kg when
betaine was used in an amount of 2.5 kg/ha. This was an
WO 96/07319 2 1 9 9 3 6 8 PCT/}i~95/00484
increase of 23.5~ over the control, i.e. about 17 t/ha.
The results are shown in Table 1.
Table 1
Effect of betaine on potato yield
betaine increase in yield
(kg/ha) (~ of the control)
0 100
1.25 112
2.50 123.5
5.00 117 5
10.00 112.5
In location (2), the results deviated to some
extent from the results obtained in location (1); an
increase of more than 10~ in the amount of yield over
the control was obtained only at the application rates
of 5 and 10 kg/ha. The best result was obtained with the
application rate of 10 kg/ha, the yield thus increasing
12.6% over the control, i.e. 7.9 t/ha. With the betaine
application rate of 10 kg/ha, a clear increase was also
detected in the number of marketable tubers per plant.
No significant differences were found in the specific
gravity of tubers. The values varied between 1.084 and
1.082.
A clear increase in the yield was apparent in
both locations in response to exogenous betaine
application. However, the yield increase was clearly
different in the two locations. The differences may
result from two different factors. On the one hand, the
stress was different in the locations due to differences
in the climate. On the other hand, in location (1) the
potato tubers were harvested within one week of the
second betaine application, and the second betaine
WO96/07319 ~ 2 1 9 9 3 6 8 PCT~5/00484 ~J
application may not have had any influence on the yield.
In location (2) betaine was added during the tuber
development stage, and the harvesting was performed at
maturity about 6 weeks after the application. The
results may thus indicate that it is inadvisable to
perform the second betaine application within a short
span of time.
Example 2
The effect of betaine on potato yield under
water stress was determined under field conditions
according to the following test arrangement:
1). Norma; irrigation (irrigation every 7 days) (WW)
2). Water stress (irrigation every 15 days) (SS)
3). Water stress (irrigation every 15 days)
+betaine (SB)
Betaine was added during the flowering of the
plants six weeks after planting. The concentration was
0.2M betaine aqueous solution. The solution was sprayed
at the rate of moisturizing but not soaking the plants
(about 20 ml/plant, i.e. 0.47 g/plant). The potato
cultivar was Alpha. The potato was grown in plots of 4.0
x 2.8 m, and the crop was harvested from plots of about
3.0 x 2.1 m. The cultivation proceeded according to
normal practice, i.e. fertilizers, insecticides and
other pesticides were added, the potatoes were earthed
up, etc. The growing time was normal for the location
used. The potatoes were harvested llO days after
planting. The results are shown in Table 2.
.' 'I '- ';'' . ~t 21 99368
- WO96107319 PCT ~ s~ao48
Table 2
Effect of betaine on potato yield under dry
conditions
Treatment
Parallel tests(WW) (SS) (SB)
1 16.6~ 11.16 10.84
11 19.70 11.94 13.70
111 20.66 7.02 14.20
Average 19.01 10.04 12.91
Water stress thus clearly decreased the yield
of tubers. On the other hand, the betaine treatment
increased the number of tubers considerably under stress
conditions. Betaine application performed in a single
dose increased the yield of tubers of water-stressed
potatoes by 30~.
Example 3
The effect of betaine on tomato yield under
water stress was determined under field conditions
according to the following test arrangement:
1). Normal irrigation (irrigation every 7 days) (WW)
2). Normal irrigation (irrigation every 7 days)
+ betaine (WB)
3). Water stress (irrigation every 15 days) (SS)
4). Water stress (irrigation every 15 days)
+ betaine (SB)
Betaine was added during the flowering of the
plants six weeks after planting. The concentration was
0.2M betaine aqueous solution. The solution was sprayed
at a rate of moisturizing but not soaking the plants
(about 20 ml/p-lant, i.e. about 0.47 g/plant). The amount
of water added was not essential for the plant's need
WO96/07319 ~ 2 1 9 9 3 6 8 PCT~S/00~8~ ~
for water. The cultivation proceeded according to normal
practice, i.e. fertilizers, insecticides and other
pesticides were added, etc. The growing time was normal
for the location used. The results of the tomato
cultivation were determined always for ten plants, and
the ripe crop was harvested manually during five weeks,
beginning eleven weeks after planting. The results are
shown in Table 3.
Table 3
Effect of betaine on tomato yield under
different conditions
Treatment
Parallel (WW) (WB) (SS) (SB)
tests
Average 379 782 492 1,221
Water stress did not produce considerable
changes in the tomato yield, on the contrary the
stressed plants provided a yield that was 30% larger
than that of the normally irrigated plants. On the other
hand, the betaine treatment increased the tomato yield
considerably; the fruit production of stressed plants
more than doubled, and the production of plants growing
under normal conditions more or less doubled.
Example 4
The effect of betaine, applied at various
stages of growth, on the development and yield of
tomatoes was determined as follows. Tomatoes, cv
Pacesetter, were planted in single rows 1.5 metres
apart. Each row was on a bed ca. 90 cm wide by ca. 20
cm high. The rows were flood irrigated at the fre~uency
of 7 to 14 days during November, 7 to lO days during
~ WO96/07319 2 1 9 9 3 6 8 PCT~5/00184
December and 5 to 9 days from the start of January
until 10 days before harvest. The soil was grey clay
loam, pH 5.7.
The trial was laid out as a randomized
complete block experiment with 6 blocks, each
containiny l replicate of each treatment. Nineteen
untreated control plots were strategically located so
that each treated plot was no more than 2 plots removed
from an untreated control.
The trial occupied an area 10 rows wide (15
metres) by 88 metres long. This was divided into 11
banks of lO plots, ezch 8 metres of row. ~efore the
treatments -were allocated, each plot was classified
according to continuity of the row of plants. Plots
with a gap larger than 0.5 metres in length were
discarded. The remaining 91 plots were classified into
plots with a continuous plant row and plots with l or
2 gaps. There were sufficient plots in the first
category for 4 blocks and sufficient in the other two
categories for 1 block of each.
The betaine glycine used was food grade
material, Lot No. 64093334 (21/10/94) and it was
supplied by Tall Bennet (Rural) P/L.
The betaine was weighed into 50.0 g and 25.0
g lots and stored in press-capped 250 ml PET
containers. Within 1 hour before use, sufficient of
these for the application were dissolved in measured
volumes of clean water to produce a 200 g/l solution.
These solutions were prepared in 250, 500 or 1000 ml
glass-calibrated cylinders and transferred to a 1 litre
screw-capped glass bottle. Within 1 hour before use,
25.0 ml of Plus-50 Surfactant (Ciba Geigy) was made up
to 250.0 ml with clean water and transferred to a 250
ml screw-capped glass bottle.
WO96/07319 ~ f 2 1 9 9 3 6 8 PCT~5/00484 ~J
14
Depending on the spray volume and swath, 1000,
1500 ml or 2000 ml of spray was prepared for
applications 1, 2 and 3 respectively. A 1000 ml glass
measuring cylinder with 25 ml graduations was used at
all applications. The required volume of the 200 g/l
betaine solution was placed in the cylinder and it was
then approximately half filled with water. The required
volume of the 10% Spray Plus-50 solution was then added
and the cylinder was filled to 1000 ml with water. This
mixture was then transferred to a clean PET sprayer
bottle with the aid of a funnel. At application 2 a
further 500 ml of water was added and at applicaticn 3
a further 1000 ml. These volumes were measured using the
original 1000 ml cylinder and transferred to the sprayer
bottle using the original funnel. The spray was
thoroughly mixed by vigorously shaking the PET sprayer
bottle. The preparation and application of each spray
was accomplished in less than 15 minutes.
Application 1 was made on day 0, at very early
flowering. At this stage, the crop was healthy and no
stress or pests were evident. Application 2 was made on
day 20 at a mid to late flowering stage of the crop,
that still was healthy with no considerable signs of
stress. Application 3 was made on day 41, the crop being
at a very late flowering stage and low levels of Big
Bud., Helicoverpa spp., Tetranychus urticae and
Nightshade being observed.
Four doses of betaine were applied in this
trial, 1.0, 2.5, 5.0 and 10.0 kg/ha.
At application 1 they were designated treatments A, B,
C and D respectively.
At application 2 they were designated treatments E, F,
G and H respectively.
At application 3 they were designated treatments I, J,
K and L respectively.
~ Wo96107319 2 1 9 9 3 6 8 PCT ~ S/0~484
Devrinol (napropamide 500 g~kg WP) was applied
pre-plant at 6.7 kg/ha for weed control. Throughout the
trial a routine programme of fungicides (mancozeb,
J. cupric hydroxide and sulphur) and insecticides
~dimethoate and esfenvalerate) was applied. The ripening
agent Etherel (400 g/l ethepon) was applied at ca l l/ha
on day 75 of this trial. The crop was well supplied with
nutrients and it was irrigated frequently. There was
never any indication that the crop was subject to more
than low levels of stress.
Immediately prior to the first application, all
plots were examined and classified acoo~~di~1g to the
continuity of the row in order to block the trial.
Additionally, to determine the plant population and crop
stage, the plants and plants with flowers were counted
in the central 6 metre length of row in each of 12
plots. These plots were selected at random from those
that had a continuous plant row.
About l hour after application, the height of
the crop at each of lO locations per plot was measured,
i.e. at points about 0.8 m apart, starting about 0.8 in
from one end. These measurements were repeated 48 and
61 days after the first application (day 48 and day 61).
On day 47 the buds, flowers and fruit on each
of lO plants per plot were counted. The plants selected
were at about the same points used f3r height
measurements. It was intended to repeat this assessment
at day 60. However, the plants could not be separated
without damaging their neighbours, since they were now
intertwined. Hence the buds, flowers and fruit in two
40 cm lengths of row were counted. Based on the average
plant population of 12 per metre of row this was about
r the equivalent of lO plants. Each 40 cm length of row
was centred on a point l.5 metres in from the relevant
end of each plot.
21 99368
WO96/07319 PCT~5/00484
16
On day 60 the proportion of red and green fruit
was estimated by counting them in 2 partings of row per
plot.
Over days 82 and 83 the red and green fruit in
a continuous 2 metre length of plant row per plot were
counted and weighed.
In 14 plots there were large Nightshade and
Wireweed plants. In these instances two 1 metre lengths
of row free of weeds and in the centre of ca 2 metres
of continuous row were selected.
The 2 or 1 metre lengths of row were measured
and the boundaries wçre cut through to the soil surface
with a cane knife. With the 2 metre lengths, cut fruit
were discarded. With the 1 metre lengths, the cut fruit
from the end nearest an end of the plot were discarded.
The cut fruit from the other end were included in the
sample and counted as half fruit.
On day 82, before the harvest assessment
started, the proportion of foliage damaged by
Tetranychus urticae was estimated in lO quadrats per
plot. Each quadrat was 0.5 metres of row and in most
plots these were placed edge to edge along the central
5 metres of row. Deviations were necessary where there
were gaps in the row or large weeds.
The betaine treatment led to a significant
increase in the height of the tomato plants between Day
O and Day 48, due to growth of the plants, and a
significant decrease between Day 48 and Day 61. The
decrease was doubtless due to the increasing weight of
the fruit the plants were bearing.
The betaine treatments had no significant
effect on the numbers of buds, flowers or fruit at Day
47 or on the numbers of buds and flowers at Day 61.
However, at Day 61 the numbers of fruit varied
significantly between treatments as a result of a
~- WO96/07319 2 1 9 9 3 6 8 PCT~5/00~8~
response to the dose but not to the time of betaine
application.
Betaine at 1.0, 2.5 and 10.0 kg/ha
significantly increased the yield of red fruit in terms
of weight and this appeared to be due to an increase in
both numbers and weight per fruit. The yield of the
betaine at 5.0 kg/ha was not significantly different
from the untreated but was lower than the yield of the
other betaine treatments.
The response of the green fruit at Day 82 to
the dose of betaine was similar to the response of red
fruit. Rut, whereas 1.0 kg betaine/ha produced the
greatest increase in the yield of red fruit, 2.5 kg
betaine/ha produced the greatest increase in the yield
of green fruit. Further, the proportional increase in
the yield of green fruit, 109~, was significantly
greater than the proportional increase in the yield of
red fruit, 13~. The large difference between the effect
of betaine on the yield of red and green tomatoes
indicated there is a potential to obtain greater
increases in the yield of red tomatoes. This is
considered most probable in crops that are more stressed
than was the crop in this trial.
The results are shown in Table 4.
WO 96107319 ''~ ; 2 1 9 9 3 6 8 PCT/FI95100484
18
v ~ a~ o N C~
.~ 'I~
1-- ~ ,~ O '~0 N N 0~ ~ eo CO
.:: Z
U~ ~,
N ~ ~ ~n ~ N ~ i~ N 0 r~) 1
~.
C~ ._ ~ D) N ~r~ N a~ ~' ~ N N N ~' -- t-- --
~ ~ ,~ ~ ~ N _ N O O r~l _ N t~l r.7 ~O
~ E
U~
1~ 0 0 N ~ O r~
._ V ~ ~:t t~J N ~i M l~J ~ N N N N N
O ~ j3 U~ O ~D 'O N o o~ Ul oN
O E
0 U~
N ~ ~ N U~ O O 0 0 ~ ~ ~ ~ ~
~ C~ ~I N O _ ~ ~t _ ~ ~ N ~ Ir~ _
O ~0 ~ ~
E ~ o o o o O O O o _ _ _ _
O
,r ~ t ~n 1,~ Q UJ ~ C~ ~ Y J X
E~
2 1 99368
WO96/07319 PCT~Sl00484
19
Under the conditions used, i.e. low levels of
, stress, the effect of betaine on yield hence varied si-
gnificantly with dose but not with time of application.
Betaine at 1.0, 2.5 and 10.~ kg/ha increased the yield
of red tomatoes, while 5.0 kg/ha reduced the yield. The
lowest dose gave the greatest increase in yield, 13~.
However, 2.5 kg/ha gave the greatest increase in the
yield of green fruit, lO9~. Totally, the yield was of
the order of 80 tonnes per hectare, about 60% more than
a well grown commercial crop of this variety is expected
to yield.
Betaine applied at mid and late flowering sig-
nificantly reduced the proportion of foliage exhibiting
symptoms of damage by Tetranychus urticae at harvest.
The reductions were ll~ and 27%, respectively and the
trend suggested that higher levels of performance are
probable. This therapeutic property of betaine is likely
to be useful in IPM programmes.
Example 5
This experiment ~X~m; ned whether betaine can
be used to protect plants from damage caused by
herbicides. The experiment was conducted under field
conditions, and metribuzin and cyanazine (Bladex) were
used as herbicides and added at a late stage of growing.
Five different concentrations of betaine were used: 0
(control), 2, 4, 8 and 12 kg of betaine per hectare. For
the purpose of dosage, an aqueous solution of betaine
was prepared, and in addition to the desired betaine
content the solution contained 1 ml/l of non-ionic
wetter, Plus-50 (Ciba Geigy). Betaine solution was added
in an amount of 640 l/ha at 25% ground cover. The potato
cultivar was Russet Burbank. The location situated at
an altitude of 140 m and was periodically plagued by
high temperatures and drought. The crop was harvested
manually, and the tubers were graded into unmarketable
WO96/07319 ~1 qq 3 6 8 PCT~5/00484
~small, green and diseased tubers) and marketable ones,
and the weight and number of tubers in the categories
were determined.
In this experiment too, betaine increased the
number of tubers. The smallest betaine application
rates, 2 to 4 kg/ha, had no significant effect on the
yield and the number of tubers. With the highest betaine
concentrations the yield and the number of tubers were
significantly increased. The number of tubers per
hectare increased the most with the betaine
concentration of 8 kg/ha, the increase being 21% over
the control. The results are shown in Table 5.
Table 5
Effect of betaine on the yield of herbicide-
treated potato
Betaine number of tubers
(kg/ha) 3
per hectare x 10 % of the control
0 170 100
2 160 94
4 176 103
8 206 121
12 181 106
