U.S. patent application number 11/808867 was filed with the patent office on 2007-12-20 for carotenoids rich paprika cultivars.
This patent application is currently assigned to The State of Israel, Ministry of Agriculture & Rural Development, A.R.O., Volcani Center. Invention is credited to Michal Barzilai, Joseph Kanner, Arieh Levy, Ezra Menagem.
Application Number | 20070294793 11/808867 |
Document ID | / |
Family ID | 33540189 |
Filed Date | 2007-12-20 |
United States Patent
Application |
20070294793 |
Kind Code |
A1 |
Levy; Arieh ; et
al. |
December 20, 2007 |
Carotenoids rich paprika cultivars
Abstract
Paprika cultivars being characterized by fruits having total
carotenoids content higher than 10 mg/g dry fruit weight and a
branching pattern suitable for mechanical harvesting and methods
and systems generating same.
Inventors: |
Levy; Arieh; (Rehovot,
IL) ; Menagem; Ezra; (Rehovot, IL) ; Kanner;
Joseph; (Rechovot, IL) ; Barzilai; Michal;
(Rehovot, IL) |
Correspondence
Address: |
Martin D. Moynihan;PRTSI, Inc.
P.O. Box 16446
Arlington
VA
22215
US
|
Assignee: |
The State of Israel, Ministry of
Agriculture & Rural Development, A.R.O., Volcani Center
Beit-Dagan
IL
|
Family ID: |
33540189 |
Appl. No.: |
11/808867 |
Filed: |
June 13, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11300363 |
Dec 15, 2005 |
|
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11808867 |
Jun 13, 2007 |
|
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10607103 |
Jun 27, 2003 |
7074990 |
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11300363 |
Dec 15, 2005 |
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Current U.S.
Class: |
800/317.1 ;
426/540 |
Current CPC
Class: |
A23L 5/44 20160801; A01H
5/08 20130101 |
Class at
Publication: |
800/317.1 ;
426/540 |
International
Class: |
A01H 5/00 20060101
A01H005/00; A23L 1/27 20060101 A23L001/27 |
Claims
1. A method of producing a carotenoid-rich composition from paprika
fruit, the method comprising: (a) obtaining at least one paprika
fruit from paprika plant Capsicum annuum cv. 1056 or 1057; and (b)
extracting the carotenoid from said paprika plant, thus producing a
carotenoid-rich composition.
2. The method of claim 1, further comprising a step of
freeze-drying said at least one paprika fruit following step (a)
and before step (b).
3. The method of claim 1, further comprising a step of grinding
said at least one paprika fruit following step (a) and before step
(b).
4. The method of claim 2, further comprising a step of grinding
said at least one paprika fruit following said freeze-drying and
before step (b).
5. The method of claim 1, wherein said extracting is acetone
extraction.
6. The method of claim 1, further comprising a step of
chromatographically purifying said carotenoid composition.
7. A carotenoid-rich composition produced by the method of claim 1.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 11/300,363 filed on Dec. 15, 2005, which is a
Divisional of U.S. patent application Ser. No. 10/607,103, filed on
Jun. 27, 2003, now U.S. Pat. No. 7,074,990, issued Jul. 11, 2006.
The contents of the above Applications are incorporated herein by
reference.
FIELD AND BACKGROUND OF THE INVENTION
[0002] The present invention relates to cultivars of paprika rich
in carotenoids and, more particularly, to carotenoids rich
cultivars of Capsicum annuum which produce high fruit yields and
are adapted to mechanical harvest.
[0003] Paprika cultivars are used commercially to produce powders,
oleoresins and concentrates as food colorants. These products are
typically very rich in carotenoids. Carotenoids are isoprenoid
compounds, with an extensive conjugated double bond system. They
are divided into two main classes; carotenes [acyclic (lycopene)
and cyclic (beta.-carotene)], and xanthophylls (e.g., capsanthin)].
In contrast to carotenes, which are pure polyene hydrocarbons,
xanthophylls also contain hydroxy, epoxy and keto groups. The
carotenoids form a group of pigments which have a yellow or red
color and confer the characteristic colors on many food and
cosmetic stuffs. Epidemiological studies have moreover shown that
frequent and regular consumption of carotenoids reduces the risk of
chronic disorders, including cardiovascular disorders, and has a
beneficial effect on cancer prevention. This protective function of
the carotenoids is seen both in their action as antioxidants and,
as in the case of beta-carotene, in their provitamin A activity
[see Journal of the American Dietetic Association, 97: 991-996
(1997)]. In particular, the mixture of these three carotenoids
represents a system with particular antioxidant properties.
Accordingly, carotenoids are highly sought for due to their
nutritive and medicinal values.
[0004] Paprika carotenoids include beta-carotene, zeaxanthin,
lutein, capsanthin, capsorubin and cryptocapsin, which confer
substantial nutritional and medical values. Epidemiological studies
have shown that frequent and regular consumption of carotenoids,
such as paprika carotenoids, reduces risks of chronic disorders,
such as cardiovascular disorders (Kohlmeier L et al. 1995. Am. J.
Clin. Nutr. 62: 137-146) or cancer (Murakoshi et al., 1992.Cancer
Res. 52: 6583-6587; Levy et al.1995. Nutr. Cancer 24: 257-267; and
Tanaka et al., 1994. Carcinogenesis 15: 15-19).
[0005] The total carotenoids content in most known commercial
paprika cultivars varies from 2 to 8 milligram per gram dry fruit
weight (Govindarajan,1986. Crit. Rev. Food Sci. Nutr. 24:245-355;
and Mingez-Mosquera et al. 1992. J. Agric. Food Chem.
40:2384-2388). In comparison, rich carotenoids tomato cultivars
contain less than 0.15 milligram carotenoids per gram dry fruit
weight. These substantial differences in carotenoids content
emphasizes the high value the paprika crop as a preferred source
for carotenoids being used by nutritional, nutraceutical, and
pharmaceutical industries.
[0006] While reducing the present invention to practice, the
present inventors have generated new cultivars of paprika which are
characterized by being highly rich in carotenoids. In addition,
these cultivars produce high fruit yields and are adapted to
mechanical harvest. Thus, the paprika cultivars of this invention
represent a valuable commercial source for nutritive and/or
medicinal carotenoids, as well as to carotenoids used in the dye
industry.
SUMMARY OF THE INVENTION
[0007] According to one aspect of the present invention there is
provided a paprika plant, or its parts, which is being
characterized by fruits having total carotenoids content higher
than 10 mg/g dry fruit weight and a branching pattern suitable for
mechanical harvesting.
[0008] According to another aspect of the present invention there
is provided a seed of the paprika plant of the present
invention.
[0009] According to yet another aspect of the present invention
there is provided a tissue culture of regenerable cells of the
paprika plant of the present invention.
[0010] According to still another aspect of the present invention
there is provided a method of generating a paprika plant having a
high carotenoids content using plant breeding techniques which
employ a paprika plant, or its parts, as a source of plant breeding
material, the method comprising utilizing paprika plants Capsicum
annuum cv. Lehava and Capsicum annuum line 4126 as a source of
breeding material.
[0011] According to an additional aspect of the present invention
there is provided a method of generating a paprika plant using
plant breeding techniques which employ a paprika plant, or its
parts, as a source of plant breeding material, the method
comprising utilizing paprika plant Capsicum annuum cv. 1056, (ATCC
Accession No: PTA-5147) or Capsicum annuum cv. 1057 (ATCC Accession
No: PTA-5148) as a source of breeding material.
[0012] According to still an additional aspect of the present
invention there is provided a system for developing a paprika plant
having a high carotenoids content using plant breeding techniques,
the system comprising paprika plants Capsicum annuum cv. Lehava,
and Capsicum annuum line 4126, or parts of the paprika plants, as a
source of the breeding material.
[0013] According to yet an additional aspect of the present
invention there is provided a system for developing a paprika plant
using plant breeding techniques, the system comprising paprika
plant Capsicum annuum cv. 1056, (ATCC Accession No: PTA-5147) or
Capsicum annuum cv. 1057 (ATCC Accession No: PTA-5148) as a source
of breeding material.
[0014] According to further features in preferred embodiments of
the invention described below, the fruit of the paprika plant of
the present invention is characterized by a beta carotene content
higher than 1.5 mg/g dry fruit weight.
[0015] According to still further features in the described
preferred embodiments the of the invention described below, the
branching pattern of the paprika plant of the present invention is
characterized by a branching angle not exceeding 40 degrees from
main stem and branching points which occurs at a height of least 30
cm above ground in mature plants.
[0016] According to still further features in the described
preferred embodiments of the invention described below, the paprika
plant of the present invention is further characterized by having
an average height exceeding the average height of a Capsicum annuum
cv. Lehava plant being of a similar age and grown under similar
conditions.
[0017] According to still further features in the described
preferred embodiments of the invention described below, the paprika
plant of the present invention is further characterized by having a
number of fruits per plant exceeding the number of fruits per plant
of a Capsicum annuum cv. Lehava plant being of a similar age and
grown under similar conditions.
[0018] According to still further features in the described
preferred embodiments of the invention described below, the paprika
plant of the present invention is further characterized by having a
dry fruit yield exceeding the dry fruit yield of a Capsicum annuum
cv. Lehava plant being of a similar age and grown under similar
conditions.
[0019] According to still further features in the described
preferred embodiments of the invention described below, the paprika
plant of the present invention is Capsicum annuum cv. 1056,
representative seeds thereof having been deposited under ATCC
Accession No: PTA-5147.
[0020] According to still further features in the described
preferred embodiments of the invention described below, the paprika
plant of the present invention is Capsicum annuum cv. 1057,
representative seeds thereof having been deposited under ATCC
Accession No: PTA-5148.
[0021] According to still further features in the described
preferred embodiments of the invention described below the paprika
plant of the present invention is further characterized at maturity
by at least one trait selected from the group consisting of plant
height exceeding 90 cm, an average fruit length of at least 11 cm,
an average fruit width of at least 2.9 cm, an average fruit dry
weight of at least 3.5 g, an average number of fruits per plant of
at least 11.7 fruits and an average fruit dry weight yield of at
least 0.65 kg per m.sup.2.
[0022] According to still further features in the described
preferred embodiments of the invention described below the paprika
plant of the present invention is further characterized at maturity
by having light brown seeds.
[0023] According to still further features in the described
preferred embodiments of the invention described below the paprika
plant of the present invention is further characterized at maturity
by having light yellow seeds.
[0024] According to still further features in the described
preferred embodiments of the invention described below, the tissue
culture of regenerable cells of the paprika plant of the present
invention is capable of expressing all the morphological and
physiological characteristics of the paprika plant.
[0025] According to still further features in the described
preferred embodiments of the invention described below, the tissue
culture of regenerable cells of the paprika plant of the present
invention is regenerated from cells or protoplasts of a tissue
selected from the group consisting of seeds, leaves, stems,
pollens, roots, root tips, anthers, ovules, petals, flowers,
embryos, fibers and bolls.
[0026] According to still further features in the described
preferred embodiments of the invention described below, the method
of generating a paprika plant having high carotenoids content is
effected by using plant breeding techniques which are selected from
the group consisting of recurrent selection, backcrossing, pedigree
breeding, restriction fragment length polymorphism enhanced
selection, genetic marker enhanced selection, and
transformation.
[0027] The present invention successfully addresses the
shortcomings of the presently known configurations by providing
carotenoids rich paprika cultivars which produce high fruit yields
and are amenable to mechanical harvest and methods and systems
generating same.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0028] The present invention is of paprika (Capsicum annuum)
cultivars which are characterized primarily by exceptionally high
carotenoids content and which are also characterized by high fruit
yields and morphological suitability for mechanical harvesting.
[0029] The principles and operation of the present invention may be
better understood with reference to the accompanying
descriptions.
[0030] Before explaining at least one embodiment of the invention
in detail, it is to be understood that the invention is not limited
in its application to the details set forth in the following
description or exemplified by the Examples. The invention is
capable of other embodiments or of being practiced or carried out
in various ways. Also, it is to be understood that the phraseology
and terminology employed herein is for the purpose of description
and should not be regarded as limiting.
Terminology
[0031] As used herein the term "line" refers to the genetic
complement contained in the plant.
[0032] As used herein the phrase "breeding line" refers to a
homozygous plant line which can be used to breed a specific
cultivar.
[0033] As used herein the phrase "homozygous plant" refers to a
plant in which both sets of chromosomes contain essentially
identical alleles in all locations.
[0034] As used herein the term "cultivar" refers to a plant of a
specific line which results from a selective breeding and
maintained by cultivation, and with characteristics that make it
unique, such as morphological characteristics or content of
specific molecules (e.g., carotenoids).
[0035] As used herein the phrase "full maturity" refers to a growth
stage wherein fruits are dark red in color and exhibit loss of
turgor.
[0036] Paprika is an important and valuable field crop. Thus, a
primary goal of paprika breeding is to select and develop plants
that have the traits that result in superior varieties.
[0037] As is illustrated in the Examples section which follows, the
present inventors have generated paprika plant breeding lines which
are characterized by a combination of traits which makes such
plants highly suitable for use as a source of carotenoids for the
food, drug and dye industries.
[0038] Thus, according to one aspect of this invention there is
provided a paprika plant which is characterized by fruit having
total carotenoids content higher than 10 mg/g dry fruit weight
(dfw) and a branching pattern and fruits suitable for mechanical
harvesting. A branching pattern suitable for mechanical harvesting
is typically characterized by branching which starts at least 30 cm
above ground and at an angle not exceeding 40 degrees from the main
stem. Plants exhibiting such branching patterns create compact
non-profuse foliage which is compatible to the limited capacity of
a standard paprika harvester; moreover, the fruits should be easily
detachable from stems.
[0039] As is further illustrated in Example 3 of the Examples
section which follows, analysis of the fruit of the paprika plant
of the present invention, revealed that total carotenoids content
and beta carotene content are substantially higher than that of
standard commercial paprika cultivars. The paprika plants of the
present invention exhibited total carotenoids content that is at
least 60% higher than that of known commercial cultivars. In
addition, the paprika plants of the present invention exhibited
extremely high beta carotene content (higher than 1.5 mg/g dfw)
which translates into at least 200% increase of this specific
carotenoid over commercial paprika cultivars.
[0040] As is mentioned hereinabove and as is illustrated in Example
2 of the Examples section which follows, the paprika plants of the
present invention are also highly amenable to mechanical harvest
further substantiating the commercial applicability of these
cultivars. Accordingly, the mature plants compactly branch at a
height of at least 30 cm above ground and at an angle not exceeding
40 degrees from the main stem. In addition, mature fruits are
easily separated from plants. Therefore, the paprika plants of the
present invention can be harvested mechanically and thus are
suitable for large scale cultivation.
[0041] Thus, the paprika plants of the present invention are highly
suitable as a source of carotenoids. Techniques for extracting and
processing carotenoids from plants are described, for example, in
U.S. Pat. Nos. 2,412,707; 2,917,539; 3,206,316; 4,400,398;
4,680,314; 5,382,714; 5,648,564; 5,310,554; 5,962,756; and
6,380,442; and European Patent Application No. 0 242,148.
[0042] Extracted carotenoids can be utilized for nutraceutical or
pharmaceutical purposes. For example, carotenoids have been used as
a source for vitamin A (retinol) production and have been
implicated for use as cancer preventative agents [for example,
Bollag, 1979. Cancer Chemother. Pharmacol. 3:207-215; Spom et al.,
1981, In: Zedeck et al., (eds), Inhibition of Tumor Induction and
Development, pp.71-100, Plentum Publishing Corp., New York; Bertram
et al., In: M. S. Amot et al., 1982 (eds.), Molecular Interactions
of Nutrition and Cancer, pp. 315-335, Raven Press, New York; and
U.S. Pat. Nos. 5,705,180; 6,200,597; and 6,428,816]. In addition,
various forms of natural carotenoids have been used for at least a
century as food and/or feed additives in human or animal diet
and/or as food and cosmetic natural colorants, such as for example,
colorants for margarine and butter. Recently, color stabilized
carotenoid colorants have been disclosed (U.S. Pat. No.
5,079,016).
[0043] The paprika plant of the present invention is exemplified by
cultivars 1056 and 1057, representative seed thereof having been
deposited under ATCC Accession Nos: PTA-5147 and PTA-5148,
respectively.
[0044] These cultivars were generated from crossing the commercial
cultivar "Lehava" and the breeding line 4126, as described in
Example 1 of the Examples section that follows. Briefly, the
breeding line 4126 was originated from a spontaneous mutant, which
contained high carotenoids content, but had a low yielding
potential and was unsuitable for mechanical harvesting. The two
lines, 4126 and Lehava, were crossed, self pollinated and selected
over six successive generations. Subsequently, two homozygous lines
were selected and designated as cultivars 1056 and 1057. These new
cultivars were grown in the field to maturity under standard
agronomic practices and evaluated comparatively with the commercial
cultivar (Lehava) for their morphological and chemical
characteristics
[0045] The paprika plants of the present invention also have
additional morphological and physiological characteristics
including a higher height, higher number of fruits per plant and a
higher fruit yield, as compared with the standard commercial
cultivar Lehava.
[0046] The paprika cultivars described herein can be used to
generate additional cultivars which exhibit the characteristics of
the paprika plant of the present invention. Plants resultant from
crossing any of these cultivars with another plant can be utilized
in pedigree breeding, transformation and/or backcrossing to
generate additional cultivars which exhibit the characteristics of
the paprika plants of the present invention and any other desired
traits. Screening techniques employing molecular or biochemical
procedures well known in the art can be used to ensure that the
important commercial characteristics sought after are preserved in
each breeding generation.
[0047] The goal of backcrossing is to alter or substitute a single
trait or characteristic in a recurrent parental line. To accomplish
this, a single gene of the recurrent parental line is substituted
or supplemented with the desired gene from the nonrecurrent line,
while retaining essentially all of the rest of the desired genes,
and therefore the desired physiological and morphological
constitution of the original line. The choice of the particular
nonrecurrent parent will depend on the purpose of the backcross.
One of the major purposes is to add some commercially desirable,
agronomically important trait to the plant. The exact backcrossing
protocol will depend on the characteristic or trait being altered
or added to determine an appropriate testing protocol. Although
backcrossing methods are simplified when the characteristic being
transferred is a dominant allele, a recessive allele may also be
transferred. In this instance, it may be necessary to introduce a
test of the progeny to determine if the desired characteristic has
been successfully transferred. Likewise, transgenes can be
introduced into the plant using any of a variety of established
transformation methods well-known to persons skilled in the art,
such as: Gressel., 1985. Biotechnologically Conferring Herbicide
Resistance in Crops: The Present Realities, In: Molecular Form and
Function of the plant Genome, L van Vloten-Doting, (ed.), Plenum
Press, New York; Huftner, S. L., et al., 1992, Revising Oversight
of Genetically Modified Plants, Bio/Technology; Klee, H., et al.,
1989, Plant Gene Vectors and Genetic Transformation: Plant
Transformation Systems Based on the use of Agrobacterium
tumefaciens, Cell Culture and Somatic Cell Genetics of Plants; and
Koncz, C., et al. 1986, Molecular and General Genetics.
[0048] A comparison of paprika cultivars 1056 and 1057 to their
parental lines (see, Examples section), demonstrates that these
paprika plants exhibit several economically and agronomically
advantageous traits over its respective parental lines. In
particular the combination of high total carotenoids content, in
particular high beta carotene content, and growth pattern suitable
for mechanical harvesting, distinct the paprika plants of the
present invention from prior art commercial cultivars.
[0049] Once established, the paprika plants of the present
invention can be propagated from seeds or alternatively by using
tissue culturing techniques.
[0050] As used herein the phrase "tissue culture" refers to plant
cells or plant parts from which paprika plants can be generated,
including plant protoplasts, plant cali, plant clumps, and plant
cells that are intact in plants, or part of plants, such as seeds,
leaves, stems, pollens, roots, root tips, anthers, ovules, petals,
flowers, embryos, fibers and bolls.
[0051] Techniques of generating plant tissue culture and
regenerating plants from tissue culture are well known in the art.
For example, such techniques are set forth by Vasil., 1984. Cell
Culture and Somatic Cell Genetics of Plants, Vol I, II, III,
Laboratory Procedures and Their Applications, Academic Press, New
York; Green et al., 1987. Plant Tissue and Cell Culture, Academic
Press, New York; Weissbach and Weissbach. 1989. Methods for Plant
Molecular Biology, Academic Press; Gelvin et al., 1990, Plant
Molecular Biology Manual, Kluwer Academic Publishers; Evans et al.,
1983, Handbook of Plant Cell Culture, MacMillian Publishing
Company, New York; and Klee et al., 1987. Ann. Rev. of Plant Phys.
38:467-486.
[0052] The tissue culture can be generated from cells or
protoplasts of a tissue selected from the group consisting of
seeds, leaves, stems, pollens, roots, root tips, anthers, ovules,
petals, flowers, embryos, fibers and bolls.
[0053] It will be appreciated that the plant lines of the present
invention can also be used in plant breeding along with other
paprika plants in order to generate novel plant lines which exhibit
at least some of the characteristics of the paprika plants of the
present invention.
[0054] For example, Capsicum annuum cv. 1056 or 1057 can be
sexually crossed with other known paprika cultivars and the
resulting progeny screened for plants having desirable
characteristics.
[0055] Thus, the present invention provides novel paprika
cultivars, and seeds and tissue culture for generating same. This
aspect of the present invention further provides a system and
method for developing such paprika plants.
[0056] Additional objects, advantages, and novel features of the
present invention will become apparent to one ordinarily skilled in
the art upon examination of the following examples, which are not
intended to be limiting. Additionally, each of the various
embodiments and aspects of the present invention as delineated
hereinabove and as claimed in the claims section below finds
experimental support in the following examples.
EXAMPLES
[0057] Reference is now made to the following examples, which
together with the above descriptions, illustrate the invention in a
non limiting fashion.
Example 1
Generation of Paprika Cultivars 1056 and 1057
[0058] Cultivars breeding: The paprika cultivars, cv. 1056 and cv.
1057, were derived from a cross between Capsicum annuum cv. Lehava,
which is the leading commercial paprika cultivar in Israel, and a
spontaneous Capsicum annuum mutant having an exceptionally high
carotenoids content and distinctively dark red pods at maturity and
dark seeds. Following three cycles of controlled self-fertilization
of the mutant, a homozygous inbred line was obtained, denoted 4126
(Levy et al. 1995. Carotenoids pigments and beta carotenes in
paprika fruits (Capsicum spp.) with different genotypes. J. Agric.
Food. Chem. 43:362-366]. The 4126 line was characterized by
exceptionally high carotenoids content (16.6 mg/g fruit dry
weight). However, the 4126 line was also characterized by having a
relatively low fruit yield, and a relatively strong attachment of
fruit to the stem, as compared with the commercial cultivar Lehava,
making it unsuitable for mechanical harvest.
[0059] Thus, a breeding program was carried, aimed at combining the
desired traits of high carotenoids content from line 4126 with the
high yield and suitability to mechanical harvest from cultivar
Lehava. Accordingly, the cultivar Lehava and line 4126 were crossed
(F1), then self-fertilized and selected, using a pedigree breeding
method, over the F2-F6 generations. This breeding program resulted
in the development of homozygous lines which retained the desired
traits. Subsequently, two cultivars were selected from these lines
and were designated as Capsicum annuum cv. 1056 and Capsicum annuum
cv. 1057.
Example 2
Morphological Characterization of Cultivars 1056 and 1057
[0060] Field evaluation procedure: The morphological and chemical
characteristics of the novel cultivars 1056 and 1057 were evaluated
comparatively with the standard commercial cultivar Lehava.
[0061] Three randomized field plots, each 7.68 m.sup.2 in size,
were used as replicates. Seeds of the three cultivars were sown in
the spring in three rows, 50 cm apart on 1.92 m wide soil beds.
Following emergence, the seedlings were thinned to a density of 6-7
seedlings per meter. The plots were irrigated with cumulative
amount of 750 m.sup.3 of water and were fertilized with 23 and 7.5
kg of nitrogen and potassium, respectively, per 1000 m.sup.2.
[0062] Immediately following harvest, the plants were measured for
height, fruit size, number and fruit weight. In addition, fruits
were chemically analyzed for their carotenoids content.
[0063] Results: Plants of both cultivars 1056 and 1057 were taller
than those of the Lehava cultivar (by 44.0% and 25.3%,
respectively) and had a compact non-profuse branching
indistinguishable from the Lehava cultivar, and therefore were
found to be highly suitable for mechanical harvesting. Cultivar
1056 had light brown seeds, while cultivars 1057 and Lehava had
light yellow and ivory color, respectively. Further
characterization (summarized in Table 1 below), showed that the
average fruit length, width and dry weight values of cultivars 1056
and 1057 were similar or slightly lower than that of the Lehava
cultivar. On the other hand, the number of fruits per plant of
cultivars 1056 and 1057 was higher than that of the Lehava
cultivar, resulting in a substantially higher fruit dry yields per
plant (by 31.0 and 43.1%, respectively). TABLE-US-00001 TABLE 1
Morphological characteristics of the plant and fruit of cultivars
1056 and 1057 in comparison with the commercial cultivar Lehava
Plant Fruit dry No. height Fruit (cm) Ratio Fruit dry yield Fruits
Cultivar (cm) length width length/width weight(g) kg/m.sup.2 per
plant Lehava 78.2 (3.0)* 13.4 (1.0) 3.8 (0.2) 3.5 (0.1) 4.9 (0.6)
0.58 (0.17) 11.4 (2.1) cv. 1056 112.6 (6.5) 12.0 (1.0) 3.4 (0.2)
3.5 (0.4) 3.8 (0.3) 0.76 (0.11) 15.6 (3.3) cv. 1057 98.0 (7.3) 13.3
(1.0) 3.1 (0.2) 4.3 (0.4) 3.9 (0.4) 0.83 (0.10) 14.3 (2.6) *Values
are means of three replicates, in parenthesis, standard errors of
the means.
Example 3
Chemical Characterization of Cultivars 1056 and 1057
[0064] Separation and Quantification of Carotenoids from Paprika
varieties:
[0065] Paprika fruits of the cultivars described herein were
harvested at full ripe maturation, i.e., when fruits turned red and
exhibited loss of turgor. The fruit pericarp was dehydrated by
freeze-drying and ground by a coffee grinder to a fine powder. The
powder was stored in closed bottles at -20.degree. C. until used
for chemical extraction and analysis.
[0066] The content of total carotenoids was determined by
dissolving a 25 mg sample of paprika powder in 25 ml of acetone for
18 hours at room temperature in the dark. The extract solution was
then analyzed for the carotenoids pigment-color intensity of at 474
nm by a spectrophotometer (HP 8452A). The total carotenoids density
was then estimated based on capsanthin extension coefficient of
1%=1905 A.
[0067] The content of separated carotenoids was analyzed using the
procedure described by Y. Ittah et al., 1993. (Hydrolysis study of
carotenoids pigments of paprika by HPLC/photodiode array detection.
J. Agric. Food. Chem. 41: 889-901). Briefly, paprika powder (30 mg)
was suspended in a 2% BHT (butylated hydroxy toluene) solution in
7.5 ml of absolute ethanol, followed by adding 1.25 ml of a 60% KOH
aqueous solution. The suspension was then stirred under nitrogen at
37.degree. C. for 30 min and immediately thereafter chilled on ice
for 10 min. Water (5 ml) was then added to the extract followed by
repeated additions of 5 mg hexane aliquots, until no color could be
observed in the extract. The combined hexane extract was then dried
over anhydrous sodium sulfate and evaporated under nitrogen and the
dried extract was added to 1 ml of acetone (HPLC grade), passed
through a 0.2 .mu.m filter and injected into a high-performance
liquid chromatography (HPLC) column (LiChrospher.RTM. 100 reversed
phase C18 column, 250/4 mm; particle size--5 .mu.m) connected to, a
photodiode array detector (SPD-M10Avp) and a software system which
controlled all the equipment and carried out data integration and
processing (CLASS-VP, Shimadzu). HPLC conditions were as follows: a
flow rate of 1 mL/min, an injection volume of 10 .mu.l; elution was
effected using the solvent gradient of Table 2 (below), detection
was at 474 nm. TABLE-US-00002 TABLE 2 Elution gradient % water %
acetone Time, min 25 75 10 25 75 5 5 95 5 5 95 7 0 100 5 25 75 5
(M. Isabel M'ingiez-Mosquera and D'amaso Horero-M'endez. Separation
and quantification of the Carotenoid Pigments in Red Peppers,
Paprika, and Oleoresin by Reversed-Phase HPLC. J. Agri. Food. Chem.
1993, 41, 1616-20.)
[0068] Following HPLC separation, the densities of specific
carotenoids in sample extracts were estimated using the integrated
area of the compound peaks and calculated from the total
carotenoids. The ASTA value of each sample was determined by
multiplying the optical density value of I mg paprika powder in 1
ml acetone solution by 164.
[0069] As summarized in Table 3 below, the content of total
carotenoids of cultivars 1056 and 1057 was 134 and 124.4 mg/10 g
dry fruit weight (dfw, respectively). Compared with total
carotenoids content of 77.6 mg/10 g dfw characterizing the Lehava
cultivar, the cultivars of the present invention exhibited a 72.7%
(1056) and 60.3% (1057) increase in total carotenoids content.
Furthermore, the beta-carotene content of cultivars 1056 and 1057,
was 17.6 and 18.4 mg/10 g dfw, respectively, compared with just 5.5
mg/10 g dfw found in the Lehava cultivar, which translates to a
220% and 234.5% increase, respectively. The total carotenoids
content and the beta carotene content of cultivars 1056 and 1057
are among the highest reported in paprika fruits (Govindarajan,
1986. Crit. Rev. Food Sci. Nutr. 24:245-355; and Mingez-Mosquera et
al. 1992. J. Agric. Food Chem. 40:2384-2388). TABLE-US-00003 TABLE
3 Chemical characteristics of the fruits of cultivars 1056 and 1057
in comparison with the commercial cultivar Lehava. Total
carotenoids ASTA Beta-carotene Capsanthin Vitamin E Cultivar mg/10
g.d.w. Units mg/10 g.d.w. mg/10 g.d.w. mg/10 g.d.w. Lehava 77.6
(1.8) 233.1 (13.4) 5.5 (0.1) 28.0 (1.6) 3.4 (0.1) cv. 1056 134.0
(9.2) 402.5 (28.3) 17.6 (1.6) 41.7 (2.0) 3.9 (0.1) cv. 1057 124.4
(3.8) 373.6 (11.4) 18.4 (0.4) 36.5 (1.3) 3.0 (0.1) * d.w. = dry
weight ** values are means of three replication, in parenthesis the
standard errors of the means.
[0070] Thus, paprika cultivars 1056 and 1057 exhibited
substantially higher contents of total carotenoids and of beta
carotene, as compared to existing commercial cultivars (Almela et
al., 1991. J. Agric. Food Chem. 39:1606-1609; Levi et al., 1995. J.
Agric. Food Chem. 43:362-366). In addition these two new cultivars
were found to be high yielding and suitable for mechanical
harvesting. Therefore, cultivars 1056 and 1057 are superior to
existing commercial paprika cultivars, particularly as a source of
nutritional and/or medicinal carotenoids.
[0071] It is appreciated that certain features of the invention,
which are, for clarity, described in the context of separate
embodiments, may also be provided in combination in a single
embodiment. Conversely, various features of the invention, which
are, for brevity, described in the context of a single embodiment,
may also be provided separately or in any suitable
subcombination.
[0072] Although the invention has been described in conjunction
with specific embodiments thereof, it is evident that many
alternatives, modifications and variations will be apparent to
those skilled in the art. Accordingly, it is intended to embrace
all such alternatives, modifications and variations that fall
within the spirit and broad scope of the appended claims. All
publications, patents and patent applications mentioned in this
specification are herein incorporated in their entirety by
reference into the specification, to the same extent as if each
individual publication, patent or patent application was
specifically and individually indicated to be incorporated herein
by reference. In addition, citation or identification of any
reference in this application shall not be construed as an
admission that such reference is available as prior art to the
present invention.
Seed Deposit
[0073] Propagating material of the paprika plant cultivars of the
present invention is maintained by American Type Culture Collection
(Manassas, Va. 20110) since Apr. 22, 2003 under the following
depository numbers: PTA-5147 and PTA-5148. Access to this deposit
will be available during the pendency of this application to
persons determined by the Commissioner of Patents and Trademarks to
be entitled thereto under 37 CFR 1.14 and 35 USC 122. Upon
allowance of any claims in this application, all restrictions on
the availability to the public of the variety will be irrevocably
removed by affording access to a deposit of at least 2500 seeds of
germplasm of these varieties.
* * * * *