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Effect of Transplanting Date on Growth, Yield andOil Quality of Tagetes minuta L. in Mid Hill of North -Western HimalayaRakesh Kumar , K. Ramesh a , Vijaylata Pathania a & Bikram Singh aa Natural Plant Products Division, CSIR- Institute of Himalayan Bioresource Technology,(Council of Scientific and Industrial Research) , Palampur (HP) - 176 061 , IndiaPublished online: 12 Mar 2013.

To cite this article: Rakesh Kumar , K. Ramesh , Vijaylata Pathania & Bikram Singh (2012) Effect of Transplanting Date onGrowth, Yield and Oil Quality of Tagetes minuta L. in Mid Hill of North -Western Himalaya, Journal of Essential Oil BearingPlants, 15:3, 405-414, DOI: 10.1080/0972060X.2012.10644068

To link to this article: http://dx.doi.org/10.1080/0972060X.2012.10644068

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Effect of Transplanting Date on Growth, Yield and Oil Qualityof Tagetes minuta L. in Mid Hill of North -Western Himalaya

Rakesh Kumar*, K. Ramesh, Vijaylata Pathania and Bikram SinghNatural Plant Products Division, CSIR- Institute of Himalayan Bioresource Technology,

(Council of Scientific and Industrial Research), Palampur (HP) - 176 061, India

Abstract: A field investigation was conducted at CSIR- Institute of Himalayan Bioresource Technology(Council of Scientific and Industrial Research), Palampur to study the effect of transplanting date on durationof phenophases, aerial biomass partitioning and essential oil productivity in wild marigold (Tagetes minuta).Tagetes crop was transplanted on six dates commencing from April 12 to June 26, 2008 at fortnightly interval.All the phenophases were hastened when the transplanting was delayed. Plant height, leaf number and branches/plant were significantly higher in the crop which was transplanted in the month of April. Dry matter accumulationin different plant parts of tagetes was significantly higher in April 26 transplanted crop. In accordance withthese results, herb yield was significantly higher in the crop which was transplanted on April 26. Delay intransplanting significantly decreased essential oil content. Delay in transplanting also caused variation incomposition of the oil in the stem, leaf and flower. Flowers and leaves accumulated maximum essential oil.The proportion of dihydrotagetones (39.1 %) was higher in leaf oil while the presence of cis- ocimene (36.5%) and ocimenones (41.3 %) in flower oil was recorded.

Key words: Tagetes minuta, date of transplanting, growth, dihydrotagetones, ocimenes andocimenones.

IntroductionTagetes minuta L. commonly known as wild

marigold is an aromatic herb of family Asteraceaenative to Argentina, South America and introducedto several other parts of the world for medicinalpurposes. The plant has rich natural productchemistry and yields volatile essential oils thatare extensively used in the cosmetic, perfumeryand as a flavoring agent in food, beverages andas medicine 1. In addition to excellent medicinalproperties it has strong nematocidal and anti-microbial activity too 2,3. The essential oil has alsobeen reported to possess insect repellentproperties, especially against ants and Aedis

aegyptis fly 4, and has been recognized for hypo-tensive, anti-inflammatory, antimicrobial,antifungal, spasmolytic and anti-inflammatoryproperties 5,6.

Though T. minuta is native to South Americabut found to occur as weed in several countriesof the world 7, including India 8. It has beenreported from South Africa, Australia, Nigeria,India, Uruguay, East Africa (Kenya), Brazil,France, Chile, Bolivia, and the Chaco region ofParaguay 9. In India, this species finds habitat inwestern Himalaya between altitudes of 1000 to2500 m amsl. Himachal Pradesh, Jammu &Kashmir and hills of Uttrakhand are the main

Journal of Essential Oil Bearing PlantsISSN Print: 0972-060X Online: 0976-5026www.jeobp.com

*Corresponding author (Rakesh Kumar)E-mail: < rakeshkumar@ihbt.res.in > © 2012, Har Krishan Bhalla & Sons

Jeobp 15 (3) 2012 pp 405 - 414 405

Received 07 August 2011; accepted in revised form 21 December 2011

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growing regions, where it occurs in natural habitatas weed in crop fields. The essential oilpredominantly contains (Z)-β-ocimene, dihydro-tagetone, (Z)-and (E)- tagetone and (Z)- and (E)-tagetenone, limonene, O-cymene 10,11,12, most ofwhich have interesting aromatic properties 7. Dueto increasing demand of tagetes oil in recent years,there has been an increasing interest in itscultivation.

Response to various agronomic practices onessential oil content and quality of T. minuta havebeen studied earlier 13,14,15,16, but no efforts havebeen made to study the effect of time of plantingon its herb yield, oil content, partitioning of oilinto different plant parts and quality of oil. Yieldand quality of essential oil bearing plants areinfluenced by environmental and managementfactors 17. Time of planting (a non monetary input)plays a significant role in improving the yield ofmany crops and governs the crop phenologicaldevelopment and total biomass production alongwith efficient conversion of biomass in toeconomic yield 18. Therefore, an attempt has beenmade to study the essential oil composition indifferent parts of T. minuta and essential oilproduction, as affected by different dates of (trans)planting.

ExperimentalExperimental siteThe study was conducted during 2008 at

Institute of Himalayan Bioresource Technology(CSIR), Palampur (32°06’05’’ N Latitude;76°34’10’’ E Longitude) India at about 1325 mamsl. The site experiences a mean annual tempera-ture of 18° C. Rainy season accounts for 65 % ofthe total rainfall exceeding 2500 mm and isassociated with low sunshine hours. The climateis classified as sub - humid. The soil is clay loamin texture, acidic in reaction (pH 5.5), having loworganic carbon (0.75 %) and low available N(181.7 kg/ha), medium available P (22.4 kg/ha)and high available K (256.2 kg/ha).

Good quality tagetes seeds were sown at definedintervals in the nursery to suit the planting dates.Six dates of transplanting viz., April 12 (T1), April26 (T2), May 11 (T3), May 26 (T4), June 10 (T5)and June 25 (T6) corresponding to 15, 17, 19, 21,23 and 26 meteorological standard weeks were

tested in a randomized complete block design.Sixty days old seedlings of tagetes weretransplanted in a well prepared field. Threereplicates were maintained. Well decomposedfarm yard manure (FYM) @ 30 t/ha wasthoroughly mixed in the soil before transplanting.A row to row spacing of 30 cm and plant to plantspacing of 30 cm was adopted in a plot of 3.3 mX 2.4 m. Periodical observations were taken oninitiation of budding, flowering, 50 % flowering,essential oil content and its composition in plantparts. Whenever a treatment plot had attained 50% flowering, the crop was harvested and data ondifferent yield parameters was calculated. Theobservations on dry matter accumulation inleaves, stem, roots and flower were recorded atharvest by uprooting five plants from outer rowon both sides. The uprooted plant were separatedinto different parts and dried in oven at 70°C tillconstant weight. Five plants were randomlyselected in the net plot area (2.85 m X 1.95 m) forrecording data on yield attributed viz., plantheight, number of branches, leaf number fromeach plot at harvest. Fifty three plants from netplot area were harvested at 50 % flowering stageand the final yield was calculated on plot basisand was converted to q/ha.

Essential oil extractionFresh aerial parts viz. leaf, stem and flowers

were harvested from net plot area from eachtreatment of T. minuta (500 g each) were subjectedto hydrodistillation (in quadruplicate) for 4 h inan all-glass Clevenger-type apparatus (hydrodistillation). Oil content was reported at v/w basis.The oil productivity (kg/ha) was calculated bymultiplying the oil content with the biomass yieldof the respective treatments and a factor of 0.9(assuming 0.9 the specific gravity of T. minutaoil). The oil was decanted and dried overanhydrous sodium sulphate (Merck). It was storedin a dark glass bottle at 4°C prior to GC-MSanalysis.

Gas chromatography analysisGas chromatographic analysis was carried out

on a Perkin –Elmer 8500 gas chromatograph withFID detector and a DB-5 capillary column (30mx 0.25 mm; film thickness 0.25 Μm). The

Rakesh Kumar et al. / Jeobp 15 (3) 2012 405 - 414 406

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operating conditions were as follows: carrier gashelium with a flow rate of 2 ml/min, the oventemperature was programmed as follows: 60°C(4min), and then 60-220°C at 4°C/min., injectorand detector temperatures were set at 240°C.

Gas chromatography - Mass spectrometryGC-MS analysis was carried out by GC-MS

(QP2010 Shimadzu, Tokyo, Japan) equipped withAOC 20i Auto sampler and DB-5 capillarycolumn (SGE International, Ringwood, Australia)of 30 m length, 0.25 mm i.d. and 0.25 μm filmthickness. Temperature was programmed from 70to 220°C at 4°C/min, held isothermally at 70°Cand 220°C for 4 and 5 min, respectively. Massspectrometer source temperature, 200°C interfacetemperature, 220 °C; injector temperature, 220°C.Sample injection volume 2 μL (dilution: 5 μL oilin 2 ml dichloromethane, HPLC grade); split ratio1:50 and mass scan 50-600 amu. Helium was usedas a carrier gas with 1.1 ml/min flow rate.

The identification of the components was madeby the comparison of their retention indices withrespect to n-paraffin (C8 – C31) internal standards.The mass spectra and retention indices (RI) werecompared with those of NIST library 19.

Statistical analysisThe data recorded on various parameters were

subjected to statistical analysis. The experimentwas laid out in a randomized complete blockdesign with three replications. The data wasanalyzed by software SYSTAT-12 (SystatSoftware Inc.,Chicago, Illinois, USA).

Results and discussionGrowth and developmentData presented in Table 1 revealed that with

delay in transplanting, phenophases viz., days tobud initiation, 50 % budding, flower initiation and50 % flowering were found to reduce. The croptransplanted on April 12 (T1) initiated buddingon 139 days after transplanting (DAT), whereas,only 72 days for those transplanted on June 25(T6). It may be due to shortening of the vegetativephase and period between budding and floweringis curtailed due to high temperature, which mighthave shortened the phenophase duration of latetransplanted crop. Due to late transplanting, thecrop was exposed to shorter day lengths (Fig. 1),tagetes being a short day plant, flowering occurearly as the day length decreases. Highertemperature coupled with shorter photoperiod

Fig. 1. Weekly meteorological data during the growing season at Palampur, (HP) India, 2008.

Rakesh Kumar et al. / Jeobp 15 (3) 2012 405 - 414 407

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Tabl

e 1.

Eff

ect o

f tra

nspl

antin

g da

te o

n gr

owth

and

dev

elop

men

t of T

. m

inut

a

Trea

tmen

tD

ays t

aken

toPl

ant h

eigh

tN

umbe

r of

bra

nche

s/L

eaf n

umbe

r /

at h

arve

st (c

m)

plan

t at

har

vest

plan

t at h

arve

stbu

d50

%Fl

ower

50 %

initi

atio

nbu

ddin

g in

itiat

ion

flow

erin

g

Dat

e of

Tra

nspl

antin

gA

pril

1213

9.3

146.

715

7.7

165.

722

5.5

7.3

615.

5A

pril

2612

7.3

132.

714

3.6

152.

324

6.7

7.3

826.

0M

ay 1

111

4.0

117.

712

8.7

138.

723

7.9

6.2

559.

5M

ay 2

699

.310

2.3

113.

312

3.0

201.

04.

226

3.3

June

10

85.0

88.3

99.3

109.

314

0.7

1.7

110.

2Ju

ne 2

571

.775

.385

.09.

711

5.7

1.3

65.6

CD

(5%

)1.

71.

11.

20.

978

.53.

012

4.0

resulted in lesser days to different phenologicalstages in late transplanted crop as compared toearlier. Due to different dates of transplanting, thecrop is exposed to different environmentalconditions with respect to temperature andphotoperiod. Consequent upon the prevailingconditions at different phases of growth the cropsexhibit considerable variation in the durationbetween different phases of growth.

Plant height was significantly higher in the cropwhich was transplanted on April 26 as comparedto June 10 and June 25 but remained statisticallyat par with remaining dates of transplanting.Number of branches/plant was significantlyhigher in the crop which was transplanted on April12 as compared to other but remained at par withApril 26 and May 11. The crop which wastransplanted on June 10 and June 25 remained atpar with each other but recorded significantlylower number of branches than other(s). Leafnumber/plant at harvest were significantly higherin the crop which was transplanted on April 26followed by April 12, May 11, May 26, June 10and June 25, respectively. April 12 and May 11transplanted crop recorded significantly similarleaf number per plant. The sink (storage organs)capacity of a crop is determined by its vegetativegrowth. Vigorous growth is associated with highersink capacity of a crop. Plant height, number ofbranches and leaf number per plant at harvest werehighest on April 26 and decreased with delay intransplanting. The crop transplanted lateaccumulated lesser degree days and resulted inpoor growth.

Dry matter accumulationThe accumulation of carbon and crop biomass

ultimately depends on the interception of solarradiation by leaf canopy and on activephotosynthesis by the individual leaves. Drymatter accumulation in different plant parts oftagetes viz., roots, leaves, stem, flowers and totalbiomass (aboveground) was significantlyinfluenced by transplanting dates. Perusal of Table2 reveals that root weight (fresh and dry) per plantwas significantly highest in April 26 followed byMay 10, April 12, May 26, June 10 and June 25,respectively. The June 11 transplanted croprecorded significantly lower root weight as

Rakesh Kumar et al. / Jeobp 15 (3) 2012 405 - 414 408

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compared to all other treatments. Similar trendwas observed for dry matter accumulation inleaves, stem, flower and total abovegroundbiomass (fresh and dry) per plant. Fresh leafweight accumulated by April 12 and April 26transplanted crop was significantly similar. FromTable 2 it is evident that the crop transplanted inthe month of June, accumulated significantlylower dry matter in different plant parts due tolesser number of days available for cropdevelopment due to shortening of phenologicalstages (Table 1).

YieldA cursory glance of table 3 revealed that tagetes

crop transplanted on April 26 recordedsignificantly higher leaf and herb yield than otherdates of transplanting but remained at par withthe crop transplanted on May 11. Delay in

Table 2. Effect of transplanting date on dry matteraccumulation in different plant parts of T. minuta

Treatments Leaf weight Stem weight Root weight Flower weight Total weight(g/plant) (g/plant) (g/plant) (g/plant) (g/plant)

Fresh Dry Fresh Dry Fresh Dry Fresh Dry Fresh Dry

Date of TransplantingApril 12 67.7 19.9 319.5 120.2 52.2 18.4 67.0 13.9 454.3 154.0April 26 109.6 36.5 662.9 220.5 162.3 64.1 105.4 44.1 877.9 301.1May 11 92.9 26.5 360.4 138.1 93.4 32.6 98.8 27.6 552.1 192.2May 26 44.6 14.9 185.5 58.7 31.0 10.1 46.8 12.7 276.9 86.3June 10 25.0 7.8 78.4 30.1 12.1 4.4 21.3 7.3 124.6 45.1June 25 16.8 3.4 40.5 13.2 6.8 2.6 8.9 3.2 66.2 19.7CD (5%) 23.3 7.4 122.3 59.7 23.2 9.6 23.9 6.5 155.8 67.0

transplanting beyond May 11 significantlyreduced leaf and herb yields. This may beattributed to the fact that lesser dry matter wasaccumulated in different plant parts of latetransplanted crop beyond May 11. Likewise stemand flower yield was significantly highest in April26 transplanted crop than other dates oftransplanting. Crop transplanted on April 26 andMay 11 behaved statistically similar. The croptransplanted in the month of June recordedsignificantly lower leaf stem, flower and herbyield. Leaf weight (fresh and dry) of single plantwas highest in April 26 transplanting whichresulted in higher leaf, stem, flower root and totalbiomass (Table 3). Crop transplanted on April 26recorded significantly higher herb yield (635.7 q/ha). Likewise Omidbaigi et al. 20 recorded highestherb yield of (700.0 q/ha) and essential oil (1.16%) from the plots of T. minuta which were

Table 3. Effect of transplanting date on yield of T. minuta

Treatments Fresh leaf Fresh Stem Fresh Flower Total Biomassweight (q/ha) weight (q/ha) weight (q/ha) (q/ha)

Date of TransplantingApril 12 46.5 227.3 35.5 299.3April 26 74.4 450.8 110.4 635.6May 11 63.1 245.1 63.5 371.7May 26 30.3 126.2 21.1 177.6June 10 17.0 53.3 8.2 78.5June 25 11.4 27.2 4.6 43.3CD (5%) 16.1 87.5 15.8 107.7

Rakesh Kumar et al. / Jeobp 15 (3) 2012 405 - 414 409

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supplied with 200 kg N/ha. Ramesh and Singh 21

while studying the planting date effect on growth,development and essential oil productivity of wildmarigold planted from 30th July to 9th Septemberat weekly interval in mid hills of Indian westernHimalaya in T. minuta reported that planting dateshad profound influence on growth anddevelopment.

Oil contentDifferent dates of transplanting affected oil

content in different plant parts viz., leaf, stem andflower on fresh weight basis (Table 4). Oil contentwas higher in flowers followed by leaf and stemin all dates of transplanting. Oil content in flowerswas higher in the crop which was transplanted onMay 11. However, oil content in leaves washighest in April 12 transplanted crop (Fig 2).Delay in transplanting decrease the total oilcontent in leaves up to May 11 but thereafter therewas slight increase and then it became constantin June transplanted crop. Total oil contentdecrease with delay in transplanting up to May26 after that it became almost constant. Climaticor planting conditions also play an important rolefor higher content in the oil. Total oil yield washighest in early transplanted crop (Table 4).Decrease in oil yield with delay in transplantingmay be due to decline in mean temperature (Fig.

3). Concentration of stem oil was highest in earlydates of transplanting whereas, concentration ofleaf and flower oil was higher in early datessubsequently decline and again increased (Table4). Flowers and leaves contained the maximumessential oil as compared to stem. These resultswere in conformity to those of Singh et al., 22 undersub-tropical plains of north India.

Oil qualityThe composition of the essential oil and

amounts of its components in T. minuta variedwith dates of transplanting (Table 4). Analysis ofleaf, stem and flower oil of T. minuta indicatedthe presence of major chemical constituents 7, 23,

11. Dihydrotagetones, tagetone, ocimenes, andocimenone were the major chemical constituentsof oil. Flower oil was a rich source of ocimenone.Maximum concentration of ocimenone was inflower oil (36.5 %) followed by stem (22.4 %)and leaf oil (11.3 %) in April transplanted crop(Table 5). However, concentration of dihydro-tagetones and tagetones was highest in leaves andocimenone was lowest as compared to flowerswhich may be due the fact that as the crop passesfrom vegetative to reproductive stage the contentof dihydrotagetone and ocimenone changes as theplant mature. Similar results were reported by Raoet al. 24 and Chamorro et al. 25. In leaf oil,

Fig. 2. Effect of transplanting dates on oil concentration in different plant parts of T. minuta

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Fig. 3. Effect of transplanting dates on oil yield (kg/ha) of T. minuta

Table 4. Effect of date of transplanting on oil content (%)in different plant parts of T. minuta

Leaf oil (% ) Stem oil (%) Flower oil (%) Total oil (%)

Date of TransplantingApril 12 0.639 0.204 0.741 1.584April 26 0.383 0.183 0.628 1.194May 11 0.266 0.057 0.759 1.082May 26 0.390 0.035 0.678 1.103June 10 0.321 0.047 0.414 0.782June 25 0.338 0.060 0.520 0.918SEm ± 0.058 0.033 0.060 0.123

maximum concentration of dihydrotagetones(39.1 %) was observed in May 26, while in stemoil it was 36.0 % during June 10 transplantedcrop. Tagetones were highest in leaf oil (36.3 %)as compared to stem and flower oil and theirconcentration in different plant parts was highestin the crop, which was transplanted in the monthof June. Ocimenones were highest in flower oilduring all the transplanting dates as compared toleaf and stem oil. Maximum concentration ofOcimenones in flower oil (41.3 %) was observedin the crop which was transplanted during May11 whereas in leaves (20.4 %) it was highest inApril 12 transplanted crops.

The variation in oil composition of T. minutatransplanted at different time may be due todifference in harvesting stages of crop (Table

5). Kokkini 26 reported that season of collectingmay strongly affect the essential oil yield ofthe plants and the concentration of its maincomponents.

Moghaddam et al. 27 while studying thechemical composition of T. minuta oil at variousharvest times reported that budding was moresuitable time for obtaining higher oil yield butfor higher percentage of some special componentssuch as (Z)- β- ocimene, dihydro-tagetones, (Z)-ocimenone, (E)-ocimenone and (E)-tagetones fullbloom is suitable time for harvest. Verma et al. 28

observed variations in the essential oil content andcomposition of Majorana hortensis due tovariation in enzyme levels and their pool sizes inresponse to changing weather conditions duringdifferent months.

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Tabl

e 5.

Eff

ect o

f dat

es o

f tra

nspl

antin

g on

oil

com

posi

tion

of d

iffer

ent p

lant

par

ts o

f T.

min

uta

Oil

Com

poun

ds (%

)ci

s-O

cim

ene

dihy

drot

aget

one

cis-

tage

tone

tran

s-ta

geto

neTa

geto

nes

cis-

ocim

enon

etr

ans-

ocim

enon

eO

cim

enon

es

RI

MS

956

1100

1118

-12

5912

74-

Dat

es o

ftra

nspl

antin

gLe

af o

ilA

pril

1211

.53

24.1

91.

7229

.61

31.3

32.

3118

.06

20.3

7A

pril

269.

7534

.68

1.65

28.8

230

.47

2.18

11.0

413

.22

May

11

10.3

327

.89

1.75

31.8

533

.62.

5812

.73

15.3

1M

ay 2

67.

0639

.11

1.87

28.0

829

.95

1.43

9.61

11.0

4Ju

ne 1

05.

6632

.30

1.8

34.4

936

.29

2.05

12.8

714

.92

June

25

8.68

30.5

92.

7930

.06

32.8

53.

0616

.64

19.7

0St

em o

ilA

pril

1220

.31

20.1

81.

9418

.01

19.9

56.

4614

.16

20.6

2A

pril

269.

7528

.20

1.8

23.8

525

.65

3.19

12.1

215

.31

May

11

10.6

323

.01

027

.39

27.3

94.

2312

.98

17.2

1M

ay 2

617

.33

19.9

92.

1620

.27

22.4

32.

5711

.43

14.0

0Ju

ne 1

05.

0636

.01

1.74

33.1

534

.89

2.27

11.0

913

.36

June

25

25.6

118

.80

3.08

18.7

221

.84.

0217

.13

21.1

5Fl

ower

oil

Apr

il 12

27.5

62.

240.

638.

779.

405.

8535

.99

41.8

4A

pril

2636

.52

1.9

0.68

6.44

7.12

6.62

28.4

835

.10

May

11

33.2

63.

930.

557.

688.

235.

4135

.88

41.2

9M

ay 2

633

.64

3.19

1.89

10.9

712

.86

4.16

31.0

935

.25

June

10

21.0

55.

561.

2314

.94

16.1

77.

0029

.55

36.5

5Ju

ne 2

525

.56

4.15

1.64

11.9

313

.57

7.80

30.8

738

.67

RI =

rete

ntio

n in

dice

s;M

S =

iden

tific

atio

n ba

sed

on c

ompa

rison

of m

ass s

pect

ra

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ConclusionHigher oil content was supported with higher

leaf and flower biomass when the crop wastransplanted in the second fortnight of April. Thestudy clearly indicated that transplanting time hadprofound influence on growth and developmentof T. minuta as reflected from the significantvariation on growth phase duration, aerial biomasspartitioning, herb and essential oil. The essentialoil obtained from different harvest-ing times dueto difference in transplanting dates and variationin phenological stages of wild marigold showedconsiderable variation in content and composition.Based on the essential oil content, the crop shouldbe transplanted on second fortnight of April.However, the composition of T. minuta oil was

rich in different constituents when the crop wastransplanted in the first fortnight of May. Finally,considering these results it is concluded that thewild marigold should be transplanted betweensecond fortnights of April to first fortnight of Mayunder the climatic conditions of north westernHimalaya.

AcknowledgementsThe authors are grateful to the Director, IHBT,

Palampur for providing necessary facility duringthe course of study. Financial assistance fromDST, New Delhi for undertaking this study ishighly acknowledged. This is IHBT PublicationNo. 1014.

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