U.S. patent number PP16,474 [Application Number 10/749,906] was granted by the patent office on 2006-04-25 for mint plant named `cim indus`.
This patent grant is currently assigned to Council of Scientific and Industrial Research. Invention is credited to Janak Raj Bahal, Mahendra Pandurang Darokar, Soni Gupta, Suman Preet Singh Khanuja, Birendra Kumar, Raj Kishori Lal, Nirmal Kumar Patra, Togarrati Padma Priya, Ajit Kumar Shasany, Anil Kumar Singh, Virendra Kumar Singh Tomar, Rakesh Kumar Upadhyay.
United States Patent |
PP16,474 |
Khanuja , et al. |
April 25, 2006 |
Mint plant named `Cim Indus`
Abstract
The present invention relates to a new and distinct mint plant
of Mentha piperita `Cim Indus`, selected through screening, field
trial and analysis of monoterpene constituents of the essential oil
of the germplasm, possessing the characters of producing high
amount of menthofuran ranging between 22 to 30% of total oil
content, high amount pulegone ranging between 9.0 to 18% of total
oil content, with essential oil content ranging between 0.32 to
0.40% of the total oil content.
Inventors: |
Khanuja; Suman Preet Singh
(Lucknow, IN), Patra; Nirmal Kumar (Lucknow,
IN), Shasany; Ajit Kumar (Lucknow, IN),
Kumar; Birendra (Lucknow, IN), Gupta; Soni
(Lucknow, IN), Upadhyay; Rakesh Kumar (Lucknow,
IN), Priya; Togarrati Padma (Lucknow, IN),
Singh; Anil Kumar (Lucknow, IN), Darokar; Mahendra
Pandurang (Lucknow, IN), Tomar; Virendra Kumar
Singh (Lucknow, IN), Bahal; Janak Raj (Lucknow,
IN), Lal; Raj Kishori (Lucknow, IN) |
Assignee: |
Council of Scientific and
Industrial Research (New Delhi, IN)
|
Family
ID: |
34711160 |
Appl.
No.: |
10/749,906 |
Filed: |
December 29, 2003 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20050150027 P1 |
Jul 7, 2005 |
|
Current U.S.
Class: |
PLT/259 |
Current CPC
Class: |
A01H
5/12 (20130101) |
Current International
Class: |
A01H
5/00 (20060101) |
Field of
Search: |
;PLT/259 |
Primary Examiner: Grunberg; Anne Marie
Assistant Examiner: Hwu; June
Attorney, Agent or Firm: Merchant & Gould P.C.
Claims
It is claimed:
1. A new and distinct mint plant of Mentha piperita variety `Cim
Indus`, substantially as illustrated and described herein.
Description
FIELD OF THE INVENTION
Genus, Species, and Variety
The present invention relates to a new and distinct mint plant of
the genus Mentha, species piperita, designated the variety `Cim
Indus`.
This variety was selected through screening, field trial and
analysis of monoterpene constituents of the essential oil of the
germplasm. This variety possesses the characters of producing high
amount of menthofuran ranging between 22 to 30% of total oil
content, high amount pulegone ranging between 9.0 to 18% of total
oil content, with essential oil content ranging between 0.32 to
0.40% of the total oil content.
BACKGROUND OF THE INVENTION
Menthofuran (3,6-dimethyl-4,5,6,7-tetrahydrocoumarone) is one of
the major constituent for aroma of the essential oil extracted from
the leaves of Mentha piperita. Because any other compound has not
duplicated the aroma effect, menthofuran is important in the
formulation of certain standardized essential oils, such as
peppermint oil. However, menthofuran is an expensive compound of
limited availability as the plants produce 0 to 6% menthofuran
(U.S. Plant Pat. No. PP11,788). Literatures are available for the
chemical synthesis of menthofuran to substitute the naturally
available menthofuran (U.S. Pat. No. 4,240,969) to reduce cost of
production. Also the acceptability of synthetic menthofuran is a
limiting factor in determining the cost of the essential oil
mixture containing synthetic components in aroma industry.
Considering the importance of menthofuran in aroma industry under
`New Millennium Indian Technology Initiative (NMITLI) programme`
launched by Council of Scientific and Industrial Research (CSIR),
India, during 2001, a systematic approach was undertaken to
evaluate the existing germplasm of M. piperita at CIMAP and breed
for genetic enhancement towards high menthofuran biosynthesis in
the essential oil. Systematic breeding experiments to allow open
pollination followed by single seed progeny selection by chemotypic
evaluation for enhanced constituent (menthofuran) led to
development of this chemotype the variety `Cim Indus`.
OBJECTS OF THE INVENTION
The main object of the present invention is to develop a new and
distinct mint plant.
Another main object of the present invention is to develop a novel
mint plant through screening, field trial and analysis of
monoterpene constituents of the essential oil of the germplasm.
Yet another object of the present invention is to develop a plant
producing high amount of menthofuran.
Still another object of the present invention is to develop a plant
producing high amount pulegone.
Still another object of the present invention is to develop a plant
producing high herbage.
Still another object of the present invention is to develop a mint
plant showing resistance against major plant disease conditions
like leaf spot, rust, powdery mildew, lepidopteran pest Spilarctia
obliqua.
SUMMARY OF THE INVENTION
The present invention relates to a new and distinct mint plant of
Mentha piperita `Cim Indus`, selected through screening, field
trial and analysis of monoterpene constituents of the essential oil
of the germplasm, possessing the characters of producing high
amount of menthofuran ranging between 22 to 30% of total oil
content, high amount pulegone ranging between 9.0 to 18% of total
oil content, with essential oil content ranging between 0.32 to
0.40% of the total oil content.
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 shows cluster analysis for the chemotype "Cim Indus"
compared to `Kukrail`, `Tushar`, and `Pranjal`.
FIG. 2 shows the RAPD profile of the Mentha piperita genotype
CIMPA/MP20.
FIG. 3 shows a twig of CIMAP/MP20
DETAILED DESCRIPTION OF THE INVENTION
Accordingly, the present invention relates to a new and distinct
mint plant of Mentha piperita `Cim Indus`, selected through
screening, field trial and analysis of monoterpene constituents of
the essential oil of the germplasm, possessing the characters of
producing high amount of menthofuran ranging between 22 to 30% of
total oil content, high amount pulegone ranging between 9.0 to 18%
of total oil content, with essential oil content ranging between
0.32 to 0.40% of the total oil content.
The plant was developed at the CIMAP research farm, Lucknow, India
and asexually propagated through suckers for further planting in
the same farm. By appearance, this new variety is a typical plant
of Mentha piperita. The difference between this new variety and the
parent or other known varieties is in the herbage at menthofucan,
pulegone and other essential components. Thus, this is a new and
distinct mint plant of Mentha piperita variety `Cim Indus`,
selected through screening, field trial and analysis of monoterpene
constituents of the essential oil of the germplasm, possessing the
following combination of characters: a. the said plant produces
high amount of menthofuran ranging between 22 to 30% of total oil
content, b. the said plant produces high amount pulegone ranging
between 9.0 to 18% of total oil content, c. the said plant produces
essential oil content ranging between 0.32 to 0.40% of the total
oil content, d. the said plant produces herbage yield ranging
between 200-220 quintal per hectare, e. the said plant is of height
ranging between 65 to 70 cms, with plant canopy of area ranging
between 78-85 cms, f. the said plant shows resistance against leaf
spot, rust, powdery mildew, lepidopteran pest Spilarctia obliqua,
g. the said plant has Quadrangular, woody stems, of color purplish
green with R.H.S. color code of 59A, the surface texture of the
stem is hairy and rough, h. the said plant has simple, opposite,
and decussate leaves, of with an upper surface of dark green color
with color code of RHS 137A, and a lower surface of green color
with color code of RHS 137C; the surface of the leaf is rough and
hairy with purple violet colour (81 A) on lower veins; i. the said
plant has leaves of chartaceous texture, j. the said plant's leaves
has Glabrous dorsal surface, with hairy on ventral veins, k. the
said plant has leaves of ovate-elliptical shape, with serrate
margins, l. the said plant has leaves with acute-acuminate tip,
obtuse base, and broad size, m. the said plant has leaf with
petiole of length ranging between 0.5 to 0.9 cm, n. the said plant
has leaf with of area about 7.91 cm.sup.2, o. the said plant has
leaf with length ranging between 1.2 to 4.3 cm, p. the said plant
has leaf width ranging between 0.6 to 2.6 cms, q. the said plant
has inflorescence of nature terminal spike, r. the said plant has
flowers of following traits: i. arranged in whorls, ii. smooth
pedicel, iii. green color pedicel with R.H.S. code 137B, iv. calyx
is glabrous, tubular, 5-lobed, margin ciliated, yellow green with
R.H.S. color code of 146C, v. corolla is tubular, 4-lobed, with
subsequeal lobes, and is light purple tending towards white,
whitish purple: 76 D; vi. flowers are whitish purple: 76 D, vii.
anthers are four in number, exserted, grayed-red with R.H.S. code
of 181A, viii. stigma is bifid. s. the said plant is able to
produce higher herbage, menthofuran and pulegone yield per unit
area as compared to other existing improved varieties, t. the said
plant produce high menthofuran when harvested 75 days after
planting and 115 days after planting. u. the said plant produce
high pulegone when harvested 75 days after planting, v. the said
plant is able to produce higher pulegone and menthofuran due to up
regulation and thus has the potential to isolate regulatory factors
for monoterpene metabolism, and w. the said plant has distinct
molecular profile by random amplified polymorphic DNA (RAPD) using
20 random primers (OPA) distinguishing the plant from the other
existing varieties.
The flowers are arranged in whorls and the inflorescence grow which
vary in length. The flowers include: Pedicel.--1.0 to 1.5 mm in
length, Yellow green (145C). Calyx.--Four sepals, persistent, 1.0
to 2.5 mm in length, Yellow green (145B). Calyx diameter.--1 mm.
Calyx texture.--Rough. Corrolla.--Whitish purple (76D) 2.0 to 4.0
mm in length, composed of 4 petals, differentiated into tube and a
limb. Corolla texture.--Smooth. Androecium.--Anthers -- Four,
ocidimetary, white, remain inside the corolla tube.
Gynoecium.--Stigma. Bifid, bicarpellary syncarpous; Ovary superior,
deeply four-lobed, bilocular; Style gynobasic arising between the
lobes of the ovary. Stigma -- Red-Purple Group 71C. Color of
ovaries -- Yellow-Green Group 151A.
The leaves have predominantly the carvone and menthofuran smell.
This variety produces no fruit and no seeds.
The present invention is related to the development of a novel high
menthofuran and pulegone producing chemotype which can also yield
high amount of pulegone through proper harvest management. The
plant chemotype was obtained through screening of the open
pollinated seed progenies of the variety `Kukrail`. The invention
is further related to the plant producing more herb yield leading
to higher production of essential oil per unit area compared to the
seed parent variety. The selected plant possesses the property of
accumulating more menthofuran and pulegone at specific
developmental stages and hence proper management prior to
processing can yield high amount of these important phytochemicals
for industrial use. This plant is unique and clearly distinct from
all other existing varieties of Mentha piperita. The new plant type
`Cim Indus` can be propagated vegetatively through suckers and
runners for commercial cultivation.
`Kukrail` is a released variety of CIMAP which is maintained along
with the germplasm of CIMAP in the field systemically every year.
Every year in the month of October, the twigs are planted in small
sized plots (3 m.times.3 m) for generation of enough planting
material for planting in the main field during the month of
January. Open pollinated seeds are collected from different
genotypes every year and analyzed for monoterpene constituents in
the essential oil. `Cim Indus` also known as CIMAP/MP20 is such a
genotype selected from open pollinated seed progenies of the
variety `Kukrail`. Mentha piperita is propagated vegetatively
through runners. With the NMITLI initiative the runners generated
from the seed plots were planted in 5 m.times.5 m plots during the
month of January 2001, following normal agronomic practices with
the objective to screen genotypes rich in menthofuran in the
essential oil. Replicated samples from each genotypes were taken
from the field by planting multiplied runners in the month of
January 2001, 2002 and 2003 for 3 consecutive years in RBD fashion
and different growth and yield characteristics were recorded (Table
1). For yield trials the replicated plots were prepared by adding
only FYM 1.5 ton per ha. The three-year averages of herb yield,
essential oil yield and the variations in major essential oil
components are detailed below for the genotype (CIMAP/MP20 compared
to the CIMAP released varieties of Mentha piperita `Kukrail`,
`Tushar`, `Pranjal`. `Pranjal` bears the U.S. Pat. No. 14,090.
TABLE-US-00001 TABLE 1 Comparative herbage, oil, menthofuran and
pulegone yield of Mentha piperita genotypes. Genotypes CIMAP/MP20
Kukrail Tushar Pranjal Oil content (%) 0.35 0.40 0.63 0.55 Herrbage
yield (Quintal/ 206.8 123.8 190.5 123.8 hectare) Oil yield (Litre
per 72.41 49.52 119.04 68.10 hectare) Menthofuran content(%) 27.24
8.727 9.648 8.385 Calculated Menthofuran 19.72 4.32 11.484 5.710
yield (Litre per hectare) Pulegone(%) 15.405 3.032 2.355 2.783
Calculated Pulegone 11.155 1.501 2.80 1.89 yield (Litre per
hectare)
If menthofuran is aimed the genotype can yield highest amount of
natural menthofuran than any other variety released and reported
which is the case for pulegone also.
Oil Extraction and GLC Analysis
Oil samples from the field grown plants were extracted by
hydrodistillation using Clevenger's apparatus and weighed to record
the yield. Over ground shoot samples were collected from the whole
plant selected randomly from the middle of the row of each
replicated plots at different days after planting (35. 55, 75, 95,
115 days after planting). Shoots collected from individual
genotypes were bulked for each treatment plot and essential oil was
distilled from all the replicates taking 500 g of bulked shoots
containing leaves. The final analysis of all the essential oil
samples was accomplished on Varian CX-3400 using a 30 m.times.0.25
mm Supelcowax-10 column. The injector and detector temperature were
maintained at 200 and 225.degree. C. respectively, with oven
temperature programmed from 60 to 200.degree. C. at the rate of
7.degree. C. min.sup.-1 increase, with initial and final holds of 2
and 5 minutes respectively. Hydrogen gas was used as carrier at the
rate of 1 ml min.sup.-1 and an aliquot of the sample was injected
with a split ratio of 1:50. Data were processed in the electronic
integrator Varian 4400 and the identification was based on
retention time of authentic samples of 1-menthol (Takasago, Japan)
and retention indices calculations (Jennings W & Shibamoto T
(1980) Qualitative analysis of flavour and fragrance volatile by
capillary GC, Academic Press Inc., New York.).
TABLE-US-00002 Table 2 Variation in major essential oil components
of Mentha piperita genotype CIMAP/MP20 at different stages of
growth. 35 DAYS 55 DAYS 75 DAYS 95 DAYS 115 DAYS Limonene 0.6405
6.5948 5.6849 5.6283 4.484 Menthone 1.4922 48.55 2.0802 3.8298
1.9770 Menthofuan 1.4922 6.2713 23.96 3.4181 27.246 Menthol 32.5681
34.138 12.7920 28.2289 14.396 Pulegone 9.6714 0.2421 10.4367
16.8836 15.405
TABLE-US-00003 TABLE 3 Comparative monoterpene component profiles
in the essential oil of M. piperata P(20), Kukrail, Tushar,
Pranjal, 115 days after planting. CIMAP/M Components P20 Kukrail
Tushar Pranjal 1 .alpha.-Pinene 0.432 0.465 0.661 0.707 2
.beta.-Pinene 1.004 0.874 1.256 1.472 3 Sabinene 0.857 1.031 0.786
0.881 4 Myrcene 4.897 0.342 0.342 0.341 5 .alpha.-terpinene 0.081
0.244 0.089 0.069 6 Limonene 4.484 2.862 2.656 3.145 7 1,8 Cincole
8.809 4.904 5.112 5.252 8 .gamma.-Terpinene 0.769 0.226 0.244 0.248
9 p-Cymene 0.304 0.309 0.105 0.130 10 3-Octanol 0.133 0.127 0.258
0.322 11 Menthone 1.970 21.292 28.248 28.339 12 Menthofuran 27.246
8.727 9.648 8.385 13 Iso 0.556 3.963 4.410 4.084 menthone 14
Menthyl 2.323 7.857 4.768 3.799 acetate 15 Neo menthol 4.757 3.870
2.897 3.288 16 Caryophyllene 0.661 0.452 0.088 0.056 17 Pulegone
15.405 3.032 2.355 2.783 18 Menthol 14.396 28.840 26.818 26.147 19
Piperitone 1.536 2.331 1.056 1.232 20 Carvone 0.606 0.376 0.585
0.796
The genotype CIMAP/MP20 has a characteristic oil profile which
expresses differentially at different stages of growth. The
menthofuran content was found to be higher at 75 days stage which
decreased during 95 days and again increased during harvesting time
115 days. Corresponding menthol content in the essential oil
content was found to be negatively correlated to the menthofuran
content at corresponding stages of growth.
Pulegone content increased after 75 days and was stabilized after
95 to 115 days (Table 2). The comparative monoterpene profiles for
different components is presented in Table 3 during harvesting time
(115 days stage).
Trichome Analysis of the Genotype CIMAP/MP20
Monoterpenes are known to be cytotoxic to plant tissues, inhibiting
respiration and photosynthesis by drastically affecting the
mitochondria, golgi bodies etc and decreasing cell membrane
permeability (Brown J T, Hegarty P K & Charlwood B V (1987) The
toxicity of monoterpenes to plant cell cultures. Plant Science
48:195-201.). Monoterpenes are either sequestered in the plants in
specialized structures like glandular hairs in Pelargonium (Brown J
T & Charlwood B V (1986) Differentiation and monoterpene
biosynthesis in plant cell cultures. In; Morris P, Scragg A,
Stafford A and Fowler M (eds) Secondary Metabolism in Plant Cell
Cultures. Cambridge University Press, Cambridge, 1986, p.68.),
trichomes in Mentha or stored in the form of non-toxic glycoside
derivatives in vacuoles e.g. Rosa spp.
So the number of trichomes at different developmental stages of the
genotype CIMAP/MP20 and its variation in different leaves (both the
upper and lower surface) situated at different level (0 level: leaf
at the tip, 1 level: next leaf down to 0 level, 2 level: next leaf
down to 1 level, 3 level: next leaf down to 2 level, 4 level: next
leaf down to 3 level,) were characterized and finally all the
trichome at different levels of the leaves were averaged,
calculated per centimeter square leaf area. A peak trichome density
was observed from 75 to 95 days in all the leaves at different
levels except the leaf at the tip. At 0 level the leaves are at
active developmental stage which may be cause for steady rate for
trichome formation (Table 4).
TABLE-US-00004 TABLE 4 Trichome density(Trichomes/cm.sup.2) in the
leaves of the genotype CIMAP/MP20 at different developmental
stages. 35 days 55 days 75 days 95 days 115 days Levels stage stage
stage stage stage 0 1416 2816 5499 5392 5400 1 1223 2592 4373 5112
3106 2 982 1349 2443 2464 2392 3 875 1168 1813 2080 1668 4 610 752
1824 1205 1477
Taxonomic description of the peppermint plant CIMAP/MP20 are as
given below: 1. Genus: Mentha. 2. Species: piperita. 3. Family:
Lamiaceae. 4. Common name: Peppermint. 5. Plant height: 65-70 cm.
6. Plant canopy: 80-84 cm. 7. Growth habit: Herbaceous, erect and
branched. 8. Stem: Quadrangular, woody, purplish green (59A). 9.
Leaf: Simple, opposite, decussate. Colour.--Dark green (137A).
Texture.--Chartaceous. Surface.--Glabrous dorsal surface, hairy on
ventral veins. Shape.--Ovate-elliptical. Margin.--Serrate.
Tip.--Acute-acuminate. Base.--Obtuse. Size.--Broad. Petiole
length.--0.5 cm14 0.9 cm. Area.--7.91 cm.sup.2. Length.--1.2 cm-4.3
cm. Width.--0.6 cm-2.6 cm. 10. Leaf: stem ratio: 1.06. 11.
Inflorescence: Terminal spike. 12. Flowers: Arranged in whorls.
Pedicel.--Smooth, green (137B). Calyx.--Glabrous, tubular, 5-lobed,
margin ciliated, yellowgreen (146C). Corolla.--Purple, tubular,
4-lobed, lobes subequeal, white. Anthers.--Four, exserted,
Grayed-red (181A). Stigma.--Bifid.
The colour codes are in accordance with The R.H.S. Colour Chart
published by The Royal Horticultural Society, 80 Vincent Square,
London SW1P 2PE,1995. The genotype CIMAP/MP20 was named and
referred as `Cim Indus` in this specification.
DNA Isolation and PCR Amplification Reactions
DNA was isolated from leaf tissue essentially according to the
protocol described previously (Khanuja S P S, Shasany A K, Darokar
M P & Sushil Kumar (1999) Rapid Isolation of PCR Amplifiable
DNA from the Dry and Fresh Samples of Plants Producing Large
Amounts of Secondary Metabolites and Essential oils by Modified
CTAB Procedure. Plant Molecular Biology Reporter 17: 74.) and
pooled DNA (equal amount from 20 individual plants of a genotype in
a field) constituted the samples for polymerase chain reactions
(PCRs) which were carried out in 25 .mu.l volume.
A reaction tube contained 25 ng of DNA, 0.2 unit of Taq DNA
polymerase, 100 .mu.M each of dNTPs, 1.5 mM MgCl.sub.2 and 5 p mol
of decanucleotide primers. The amplifications were carried out
using the DNA Engine thermal cycler (MJ Research, USA) following
the protocol of Khanuja et al. (Khanuja S P S, Shasany A K,
Srivastava A & Sushil Kumar (2000). Assessment of genetic
relationships in Mentha species. Euphytica 111: 121-125.). The
amplified products were loaded in 1.2% agarose gel containing 0.5
.mu.g ml.sup.-1 of ethidium bromide and photographed by Polaroid
system. Twenty decamer primers procured from Operon Technologies,
USA (OPA) were used to detect polymorphism in the selected
genotype. The similarity matrix obtained after multivariant
analysis using Nei and Li's coefficient (Nei, N. & W. Li,
1979.
Mathematical model for studying genetic variation in terms of
restriction endonucleases. Proc. Natl. Acad. Sci. (USA) 76:
5269-5273.) is shown in Table 5. These similarity coefficients were
used to generate a tree for cluster analysis using UPGMA method
(FIG. 1) which shows the distinctiveness of the genotype
CIMAP/MP20. The program `NTSYS, Numerical Taxonomy System, Version
2.1, 2000` was employed to generate the cluster diagram from the
similarity indices. The genotype CIMAP/MP20 was 54.1%, 50.2% and
51.1% different with the varieties `Kukrail`, `Tushar` and
`Pranjal` respectively establishing the uniqueness of the genotype.
These primers were also used to develop a unique RAPD profile of
the chemotype `Cim Indus` (FIG. 2). The band similarities were not
derived from a photograph but were calculated directly from the
RAPD profiles from the agarose gels based on which the cluster
diagram was made.
TABLE-US-00005 TABLE 5 Similarity between the genotypes compared
through RAPD profile analysis. Kukrail Tushar Pranjal CIMAP/MP20
Kukrail 1.000 Tushar 0.960 1.000 Pranjal 0.950 0.960 1.000
CIMAP/MP20 0.459 0498 0.489 1.000
FIG. 2 illustrates the RAPD profile of the Mentha piperita genotype
CIMPA/MP20. In this Figure Lane 1 represents the .lamda. Hind III
marker and lanes 2 to 21 represent, respectively, the profile for
each of OPA 01 through OPA 20. The identities of the primers
employed in OPA 01 through OPA 20 are provided in the accompanying
sequence listing as SEQ ID NO:1 through SEQ ID NO:20,
respectively.
Uniformity and Stability
Like any other Mentha piperita genotype this genotype is also
planted vegetatively through runners and suckers. No variation of
any kind was observed in this genotype for the last 3 years of
trial maintaining the quality of oil and phenotype. The RAPD
analysis of random plant samples in different years of trial also
did not show any variation in profiles for this genotype indicating
the stability of this genotype.
Disease and Pest Resistance
The incidence of lepidopteran pest Spilarctia obliqua and fungus
mint rust (Puccinia sps), leaf spot and mildew were not detected in
the field continuously for 3 years in the genotype CIMAP/MP20.
Metabolic Regulation of Menthofuran Biosynthesis
The genotype CIMAP/MP20 was rich in pulegone and menthofuran. In
the biosynthetic pathway Geranyl pyrophosphate is converted to
limonene which in turn is transformed into isopiperitenol, followed
by pulegone. Pulegone is converted to menthone followed by menthol.
Menthol and Menthone are the main constituents of the essential oil
of Mentha piperita. In one branch of the pathway pulegone is
converted to menthofuran. In this genotype the monoterpene pulegone
is biosynthesized at an accelerated rate and the reaction favour
more towards the menthofuran synthesis than menthol. In the initial
stage (up to 55 days stage) of growth of this genotype the reaction
favours for the production of menthol at a reduced rate from
pulegone with less accumulation of pulegone and menthfuran. But at
later stage as the plant matures the reaction favours accumulation
of more menthofuran and pulegone and instead the biosynthesis of
menthol decreases. This indicate the role of regulatory proteins in
the monoterpene metabolism and the importance of this genotype for
the isolation of such protein for future modification of metabolic
pathway modification.
SEQUENCE LISTINGS
1
20110DNAArtificialOPA primer 1caggcccttc 10210DNAArtificialOPA
primer 2tgccgagctg 10310DNAArtificialOPA primer 3agtcagccac
10410DNAArtificialOPA primer 4aatcgggctg 10510DNAArtificialOPA
primer 5aggggtcttg 10610DNAArtificialOPA primer 6ggtccctgac
10710DNAArtificialOPA primer 7gaaacgggtg 10810DNAArtificialOPA
primer 8gtgacgtagg 10910DNAArtificialOPA primer 9gggtaacgcc
101010DNAArtificialOPA primer 10gtgatcgcag 101110DNAArtificialOPA
primer 11caatcgccgt 101210DNAArtificialOPA primer 12tcggcgatag
101310DNAArtificialOPA primer 13cagcacccac 101410DNAArtificialOPA
primer 14tctgtgctgg 101510DNAArtificialOPA primer 15ttccgaaccc
101610DNAArtificialOPA primer 16agccagcgaa 101710DNAArtificialOPA
primer 17gaccgcttgt 101810DNAArtificialOPA primer 18aggtgaccgt
101910DNAArtificialOPA primer 19caaacgtcgg 102010DNAArtificialOPA
primer 20gttgcgatcc 10
* * * * *