clarysage

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Production potential, nitrogen use efficiency and economics of clarysage
(Salvia sclarea L.) varieties as influenced by nitrogen levels under different
locations
M. Yaseen*, Man Singh, Dasha Ram and Kambod Singh
Central Institute of Medicinal and Aromatic Plants, Lucknow-226015, India
A B S T R A C T
Clarysage (Salvia sclarea L.) is known for its high value essential oil, widely used in perfumery
as a source of fragrance with refreshing and long lasting note. Ten treatment combinations
consisting of two clarysage varieties (Local and CIM-Chandni) and five nitrogen levels (0, 40,
80, 120 and 160 kg N ha-1) were evaluated in factorial randomized block design with three
replications at three locations; Lucknow (Sub-tropical plain), Pantnagar (Foot hills of
Himalayas) and Purara (Western Himalayas). Crop matured 9 and 27 days earlier at Lucknow as
compared to Pantnagar and Purara, respectively. Flower spike and oil yields were highest at
Pantnagar as compared to other locations. In spite of lowest yields at Purara plant synthesized
significantly more oil due to lower temperature, longer life span and higher spikes: leaf + stem
ratio. Linalyl acetate content in oil was highest at Lucknow followed by Pantnagar and Purara.
Variety CIM-Chandni had significantly higher flower spikes, spikes: leaf + stem ratio, oil
content and produced higher oil of better quality. Optimum doses of nitrogen were worked out to
be 107.6 and 116.5 kg ha-1 at Lucknow, 117.2 and 105.2 kg ha-1 at Pantnagar, 159.8 and 132.6 kg
ha-1 for local and CIM-Chandni, respectively.
Keywords: Clarysage (Salvia sclarea), CIM- Chandni, Nitrogen use efficiency, Linalyl acetate,
Linalool, Sclareol
______________________________________________________________________
•Corresponding author. Tel.: +91 9450453745; fax: +91 522 2342666
E-mail address: yaseencimap@yahoo.com, m.yaseen@cimap.res.in (M.Yaseen)
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1. Introduction
Clarysage (Salvia sclarea L., family- Lamiaceae), a plant of southern European and central
Asian origin (Simon, et al., 1984) is commercially cultivated in Russia, Bulgaria, France and
Morocco (Vasilev, 1930) with an annual world production of 1500 tons essential oil
(Anonymous 2005). In India only a few kg oil is being produced in Kashmir valley and internal
requirement of the oil is met through import. The plant is known for its high value essential oil,
widely used in perfumery industries as a source of fragrance with refreshing and long lasting
note. Besides, the oil has also got several medicinal properties and used in stress, tension,
depression, insomnia, sore throat, indigestion, menstruations, hysteria and wild colic etc.
(Grieve, 1974). It was introduced at CSIR- Central Institute of Medicinal and Aromatic Plants
(CIMAP), research farm Kashmir (India) during 1978 from Bulgaria (Husain, 1980) and oil
produced was of international standards (Tajuddin et al., 1982). In spite of favourable climatic
conditions for its growth in Kashmir, the crop could not be commercialized there because of
traditional variety which was unsynchronized in flowering and late maturing. Realizing the uses
and demand of its high value essential oil, efforts were made for its introduction in north Indian
plains and was successfully introduced and domesticated at CSIR-CIMAP, Lucknow as an
annual winter crop (Yaseen et al., 2006). However, it could not be fitted in traditional cropping
system due to late maturing nature.
Thus, an improvement programme in clarysage was started with the aim to develop a
cultivar having synchronized flowering behaviour, short duration, more stable in yield with
wider adaptation and to fit into traditional cropping system. In this series a variety CIM-Chandni
was developed (Yaseen et at., 2005). Prior to commercialization of newly developed variety
there is need to test its potential and stability in different north Indian climatic conditions in
relation to yield and quality of essential oil as the climatic conditions affect the duration of
phenophases, yield and quality of essential oil (Ilieva, 1989). Keeping above in view, field
experiments were conducted to work out the optimum dose of nitrogen for two varieties of
clarysage under three north Indian ecological conditions.
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2. Materials and methods
2.1. Experimental site
Field experiments were conducted for two consecutive years (2007-08 and 2008-09) at three
north Indian locations [L1- CIMAP, Lucknow- North Indian Plain: located at 26°5 N latitude,ꞌ
80°59 E longitude and at an altitude of 123 m above mean sea level (amsl); L2- CIMAP,ꞌ
Pantnagar- foot hills of Himalayas: located at 29°3 N latitude, 79ꞌ°31 E longitude and at an altitudeꞌ
of 241 m amsl; L3-CIMAP, Purara- Western Himalayas: located at 29°8 N latitude, 79ꞌ°8 Eꞌ
longitude and at an altitude of 1250 m amsl]. The experimental site of L1 is classified as semi-arid
sub-tropical zone with severe hot summer and fairly cool winters. In this region monsoon normally
sets from last week of June and continues till end of September with an average annual rainfall of
1000 mm. About 80% of the monsoon rains are received during July to September. Winter also
experiences some rains due to cyclonic disturbances in Arabic sea. Mean maximum and minimum
temperature fluctuated from 24.5 to 44.5ºC and 6.9 to 27.5 ºC, respectively. The temperature was
lowest during mid December to end of January and an increasing trend in mean temperature was
noticed from first week of February and reached to highest in mid May and it declines only after
the onset of rains. The soil of the experimental site was sandy-loam (Entisol) in texture having pH
7.6 and low in organic carbon (OC) 0.3% and available nitrogen (N) 140 kg ha-1, medium in
available phosphorus (P) 12.5 kg ha-1 and potassium (K) 240 kg ha-1.
Site L2 had sub-tropical humid climate with hot summers and cool winters and receives 1535
cm annual precipitation. About 75% of the monsoon rains are received during July to September.
Winter also experiences some rains. Mean maximum and minimum temperature fluctuated from
22 to 42.5 ºC and 3 to 23 ºC, respectively. The soil of the experimental site was clay-loam
(Molisol), in texture having pH 7.2 and OC 0.9 %. The status of available N, P and K were 225,
24 and 280 kg/ha-1, respectively.
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Site L3 is classified as temperate with mild summer and cool winters. In this region monsoon
normally sets from second fort night of June and continues till end of September with an average
annual rainfall of 1200 mm. Winter also experiences frost and snowfall. Mean maximum and
minimum temperature fluctuated from 19 to 34.2ºC and 1 to 17.8º C, respectively. The soil of
experimental plot was sandy – loam (Ustochrept) with pH 6.2 and OC 0.36 %. The status of
available N, P and K were 160.3, 7.5 and 130 kg ha-1, respectively.
2.2. Treatments and experimental design
Ten treatment combinations consisting of two varieties (Local and CIM-Chandni) of
clarysage and five nitrogen levels (0, 40, 80, 120 and 160 kg N ha-1) were evaluated in factorial
randomized block design with three replications at all the three locations (Lucknow, Pantnagar
and Purara).
2.3. Raising of crop
Raised nursery beds were prepared and FYM @ 2 kg m-2 was properly mixed in the soil and
seeds were sown in the end of September using 20 g seeds m-2. To ensure proper germination
and growth light irrigations were applied through sprinkler. After primary tillage operations in
main field uniform basal dose of 60 kg P2O5 and 40 kg K2O ha-1 were applied through single
super phosphate and murate of potash, respectively before transplanting. Forty days old healthy
seedlings of clarysage were transplanted in the field plots at 45 x 30 cm spacing in second week
of November 2007 and 2008. Nitrogen in the form of urea was top dressed in three equal splits
at 15, 60 and 120 days after transplanting (DAT) as per treatments. Plots were irrigated
immediately after transplanting for proper establishment of the crop. In addition to above five
irrigations each of 50 mm in depth were applied at pre-branching, branching, bud-initiation, 50%
and full blooming stages. Two manual weeding were done at 30 and 60 days after transplanting.
2.4. Plant sampling and biometric observations
For recording observations on plant height, number of branches and flower spikes per plant
and flower spikes: leaf + stem ratio, five plants were randomly selected from each plot at the
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time of harvesting (second week of April at L1, last week of April at L2 and second week of
May at L3). For the estimation of essential oil content 200 g fresh flower spikes from each plot
were hydro-distilled using Clevenger type apparatus. The oil yield was calculated by multiplying
the flower spike yield with oil content and 0.9 (approximate specific gravity of oil).
2.5. Chemical profile of essential oil
Gas chromatography (GC) analysis of essential oil samples were carried out on Nucon gas
chromatography model 5765 equipped with a flame ionization detector (FID), BP-20 (30 m x
0.25 mm x 0.25μm film thickness). The temperature was programmed from 70-230 ºC at
4°C/min with an initial and final hold tine of 2 min. The split ratio was 1:3. The injector and
detector temperature were 200 and 230 °C on BP-20; 220 and 300°C on Pe-5 column,
respectively. Gas chromatography- mass spectrometry (GC-MS) was carried out on a Perkin
Elmer Auto System XL GC and turbo Mass Spectrometry fitted with a fused silica capillary
column, PE-5 (50 m x 0.32 mm, film thickness 0.25 μm). The column temperature was
programmed at 100-280°C at 3°C/ min using helium as a carrier gas at a constant pressure of 10
psi. The injector and detector temperature were 220 and 300°C. MS conditions were EI mode 70
eV, ion source temperature 250 °C.
2.6. Statistical analysis
The data recorded at all the locations were combined and analyzed statistically using the
techniques described by Panse and Sukhatme (1985). Critical difference (CD/LSD) values at 5%
level of probability were calculated for comparing the treatment means. For the estimation of
optimum dose of N (kg ha-1) and optimum essential oil yield (kg ha-1) for both the varieties,
quadratic equations were fitted. For the calculation of optimum dose of nitrogen and optimum oil
yield, net return per rupee investment on nitrogen, the price of clarysage oil and nitrogen were
taken as Rs. 5000 and Rs. 12.50 kg-1, respectively.
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3. Results and Discussion
3.1. Flower Bud initiation and maturity studies
Clarysage planted at Purara took significantly more time in flower bud initiation and maturity
followed by Pantnagar and Lucknow (Table 1). Crop maturity was delayed at Purara due to
lower temperature prevailing during entire cropping season. Above results are in conformity to
those reported by Ilieva (1989).
Across different locations and nitrogen levels, variety CIM-Chandni flowered 8 days and
matured 10 days earlier as compared to local (Table 1). Similar observations were recorded by
Yaseen et at. (2005).
Both flowering and maturity were significantly delayed by increasing the nitrogen levels up to
80 kg ha-1, further increase in nitrogen level though delayed flowering and maturity but the
differences were not significant (Table 1). Delay in flowering by application of nitrogen was due
to increase in vegetative growth of plants.
3.2. Growth and yield attributes
3.2.1. Location
Clarysage plants had significantly maximum height and number of floral spikes plant-1 at
Pantnagar being lowest at Purara (Table 2). However, number of branches plant-1 and flower
spikes: stem + leaf ratios recorded at Purara were significantly higher than Lucknow but
remained at par with Pantnagar. Plant synthesized significantly more essential oil (0.186 %) at
Purara as compared to 0.139% at Pantnagar and 0.127% at Lucknow. This was due to lower
temperature throughout the cropping season at Purara.
3.2.2. Varieties
The data presented in table 2 indicated that variety CIM-Chandni attained significantly higher
plant height, more number of branches (6.8) and spikes (232) per plant; higher flower spikes:
leaf + stem ratio (0.57) and essential oil content (0.164 %). as compared to the local. The
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respective values for local variety were 5.6, 176, 0.45 and 0.137%, respectively. The results are
in conformity with earlier findings (Yaseen et al., 2005).
3.2. 3. Nitrogen Levels
The data presented in table 2 revealed that application of nitrogen significantly influenced
growth and yield attributing characters of clarysage. Plant height, number of branches and flower
spikes per plant increased significantly with increase in nitrogen levels up to 120 kg ha-1. Further
increase in nitrogen level beyond 120 kg ha-1 though increased the growth and yield attributes
but the differences were not significant. Flower spikes: leaf +stem ratio increased only up to 80
kg N ha-1 which declined significantly at160 kg N ha-1. Essential oil content in flower spikes
increased with increase in nitrogen level up to 80 kg ha-1 beyond which it declined significantly.
3.3. Flower spike and essential oil yields
3.3.1. Main effect
Data presented in table 3 revealed that variety CIM-Chandni produced significantly higher
fresh flower spikes (15.81t ha-1) and essential oil (22.5 kg ha-1) as compared to 13.12 t ha-1 flower
spike and 15.3 kg ha-1 essential oil in local variety due to more number of branches and spikes
per plant, higher flower spikes: leaf + stem ratio and essential oil content in spikes. The results
are in conformity with earlier findings (Yaseen et al., 2005).
Flower spike yield of both the varieties increased significantly with increase in nitrogen level
up to 120 kg ha-1 beyond which there was no change.
3.3.2. Interaction effects
Interaction between location x variety was not significant whereas the interactions between
variety x nitrogen levels for oil yield only and location x nitrogen levels for both flower spike
and oil yields were found to be significant.
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3.3.2.1. Varieties and nitrogen levels
Interaction between clarysage varieties and nitrogen levels in respect of essential oil yield was
significant (Table 3). Variety CIM-Chandni produced significantly higher oil yield than local
cultivar at all the levels of nitrogen. Essential oil yield in CIM-Chandni increased significantly
with increase in nitrogen level up to 120 kg ha-1, whereas this increase in local cultivar was
found to be significant only up to 80 kg N ha-1. The increase in essential oil yield was mainly
because of higher values of flower spikes: leaf + stem ratio and oil content in spikes.
3.3.2.2. Locations and nitrogen levels
Interaction between Locations and nitrogen levels in respect of flower spike and essential oil
yields were significant (Table 4). Flower spike yield at Lucknow increased significantly with
increase in the nitrogen up to the highest level (160 kg ha-1) whereas the response of nitrogen at
Pantnagar and Purara was recorded only up to 120 kg ha-1.
Essential oil yield at Lucknow increased with increase in nitrogen level up to 120 kg ha-1 but
the difference between 80 and 120 kg N ha-1 were found to be non significant. At Pantnagar oil
yield increased significantly with increase in nitrogen level up to 80 kg ha-1, whereas at Purara
the increase was recorded up to 120 kg N ha-1. Similar trends were observed by Singh et al.
(2009).
3.4. Chemical profile of essential oil
Out of 43 constituents showed in GC/MS only 23 could be identified which accounted for
97% of the essential oil, the rest of the components produced at trace level could not generate
proper MS. Here, we are discussing only three major constituents: linalyl acetate, linalool and
sclareol and depicted in fig 1.
Clarysage grown at Lucknow contained higher linalyl acetate in essential oil of both the
varieties followed by Pantnagar and Purara and reverse trend was noted for linalool and sclareol
contents. Similar results were reported by Yadav et al. (2010).
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Essential oil of CIM – Chandni had higher linalyl acetate at all the locations (Lucknow-52.5,
Pantnagar-50.2, Purara- 45.2%) as compared to local variety (Lucknow - 48.9, Pantnagar- 47.3,
Purara -37.5%) and the reverse was true for linalool and sclareol contents.
Linalyl acetate content in the essential oil increased and linalool content decreased with the
increase in nitrogen levels up to 80 kg ha-1 and beyond which reverse trend was noticed in both
the varieties at all the locations. Sclareol content in oil was recorded to be highest at 80 kg N ha -1
in CIM-Chandni while in local variety it was increased up to 160 kg N ha-1 level.
3.5. Response of varieties to nitrogen at different locations
Quadratic equations (Fig.2) were fitted for response of nitrogen to oil yield for both the
varieties at all the locations. From the equations, optimum doses of nitrogen were worked out to
be 107.6 and 116.5 kg ha-1 at Lucknow, 117.2 and 105.2 kg ha-1 at Pantnagar and 159.8 and
132.6 kg ha-1 at Purara for local and CIM-Chandni, respectively. Essential oil yield at optimum
doses of nitrogen were 16.4 and 28.4 kg ha-1 at Lucknow, 21.1 and 30.3 kg ha-1 at Pantnagar and
17.6 and 21.1 kg ha-1 at Purara for local and CIM-Chandni, respectively.
3.6 Nitrogen use efficiency and economics
Nitrogen use efficiency measured in terms of increase in essential oil yield with each kg increase
in nitrogen at optimum dose was highest (122.8 g) at Pantnagar followed by 113.7g at Lucknow
and lowest 80.9 g at Purara in variety. CIM-Chandni, whereas in local variety the values were
57.8, 72.2 and 63.7g, respectively. Similar trends were noted for increase in net return and net
return per rupee investment on nitrogen (Table 5).
4. Conclusion
Based on our results, it is concluded that
i) Variety CIM-Chandni was found to be superior at all the locations.
ii) Clarysage grown at Pantnagar produced maximum flower spike and essential oil.
iii) Optimum doses of nitrogen were worked out to be 107.6 and 116.5 kg ha-1 at Lucknow, 117.2
and 105.2 kg ha-1 at Pantnagar, 159.8 and 132.6 kg ha-1 for local and CIM-Chandni,
respectively.
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Acknowledgement
The authors are grateful to the Director, Central Institute of Medicinal and Aromatic Plants,
Lucknow for providing facilities.
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