annual reports 2008

Annual Report2008

Central Institute of Medicinal and Aromatic Plants

(CSIR), Lucknow

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The green path to

better health and life

CIMAPPhone : + 91- 522 - 2359623Fax : + 91- 522 - 2342666email : [email protected] website : www.cimap.res.in w

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With compliments from

DirectorCIMAP

© copyright 2008 CIMAP, Lucknow, India

CENTRAL INSTITUTE OF MEDICINAL AND AROMATIC PLANTS(Council of Scientific Industrial Research)P.O. CIMAP, LUCKNOW – 226 015, INDIA

Front Cover : Albizia lebbeck inflorescence and podsBack Cover : Albizia lebbeck treeAlbezzia lebbeck flowers and pods are important source of a synergistic formulation for controlling

lepidopteran insects in combination with Bacillus thuringiensis δ-endotoxin (CIMAP invention).

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CIMAPORGANICS

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A Team CIMAP Effort

1

Central Institute of Medicinal and Aromatic Plants

BIO-VILLAGE MISSION

Artemisia annua

Vetiver

Withania

Patchouli

Artemisia annua was successfully cultivated for the first time in south Indian agroclimatic conditions at CIMAP

Resource Centre, Bangalore and good herb yield (15-20 t/ha) and artemisinin content (0.7-1.0%) were

obtained. Artemisia biovillage programme was successfully extended to state of Tamil Nadu where 7 acre land

at a village near Salem was put under Artemisia cultivation with excellent economic yield and artemisinin

content (0.89%)

Under the Vetiver (Khus) Biovillage programme three varieties of Khus namely CIMAP/ KS-I, Gulabi and Dharini

slips were distributed among 45 selected farmers of Sitapur, Barabanki and Raebareli districts of Uttar Pradesh.

The feedback from 25 farmers for root and oil production indicated that average production of root was in the

range of 15-20 q/ ha with oil recovery of 1.0 -1 .5% on tank basis. Digging of roots of Khus is the main hindrance

and costly operation for its cultivation by the farmers. Field demonstration of root digging by J.C.B. machine

(catterpillar) for large area and modified tractor operated Khus digger for medium area was done successfully

and was found to several fold cheaper than manual digging.

Based on the agrotechnology and seeds provided by the CIMAP Resource Centre, Hyderabad a group of farmers

started cultivation of aswagandha on pilot scale in Guntakal Kottala village in Guntakal region in 2003. During

2005-06, the cultivation was taken up in relatively large area and an association of medicinal plants growers

was formed in Gooty, Aananthapur consisting mainly of small and marginal farmers (range of land holding 2 -12

ha). This has facilitated marketing of the root. Aswagandha is now cultivated by 79 farmers in 26 villages

belonging to 13 revenue mandals in 495.5 ha replacing the traditional groundnut, redgram and Bengalgram in

some villages. The largest area has been in Guntakal Kottala village (CIMAP's model biovillage) and total area

was highest in Uravakonda revenue mandal. During 2006-07, CIMAP's agrotechnology was demonstrated for

cultivation of Withania variety Poshitha. One hundred and seventy six farmers were trained at Guntakal - the

heart of Aswagandha belt in Ananthapur district.

CIMAP Resource Centre, Bangalore launched a bio-village programme in this area in September 2003. Initially

10 farmers were selected and given planting material of a superior variety (Johore) of patchouli acceptable to

the industry and other inputs for cultivation. Training programmes were conducted by CIMAP, RC, Bangalore to

educate the farmers of the region in cultivation of patchouli. Now more than 350 farmers have taken up

cultivation of patchouli covering an area of more than 300 acres. One of the farmers, who was trained by CIMAP

on distillation, has established distillation unit and also entered into a buyback agreement with the farmers of

the area. Over a period of 3 years, the returns from the patchouli cultivation have improved the rural livelihoods

of the farmers in the area through increased income from their lands. Thus an end to end mission for patchouli

has been successfully established at Siddapur in North Kanara District in Karnataka.

Aroma Biovillage

CIM- Hariyali: High essential oil yielding variety of Rosemarinus officinalis

CIM-Shakti: High yielding variety of Satawar (Asparagus racemosus) with high root yield and high saponin

content

Vetiver as annual crop

New dimensions in organic agriculture

Aroma Biovillage jointly sponsored by Department of Science and Technology and Council of Scientific and th

Industrial Research, New Delhi has been launched at CIMAP on 4 August, 2007 by Shri Kapil Sibal, Minister of

Science and Technology and Earth Sciences, Government of India. In this programme about 100 beneficiary

farmers (50 ha) of Sultanpur and Raebareli have been adopted for cultivation of mint in normal cultivated lands

and Basil, Palmarosa, Lemongrass and Khus in salt affected soils.

Rosemarinus officinalis commonly known as 'Rosemary', a member of family Lamiaceae is an aromatic plant

and its essential oil is used in high-grade perfumery, cosmetic and soaps. The variety CIM –Hariyali has 1.03% oil

with a yield potential of nearly 350 litres of oil/ ha accounting for more than Rs. 3.0 lakhs per hectare after three

years of planting.

Variety CIM-Shakti consistently showed high saponin content in all field evaluation yield trials. The average dry

root yield is about 148q/ha and saponin content 21%. The new variety is characteristically having numerous

fingers and profuse seed formation.

Vetiver is grown as a perennial crop and its harvesting is recommended 18-24 months after planting. The long

gestation period of vetiver crop is thought to be one of the reasons for its low popularity among growers. So far,

no agrotechnology on the productivity of vetiver as annual crop is available. Keeping these in view CIMAP has

developed agrotechnology to grow vetiver as annual crop for which a period of 10-12 months has been found to

be optimum for production of economic oil yield with good quality oil. It can suitably be intercropped with some

important crops like Mentha arvensis, M. piperita and Kalmegh which could generate additional revenue.

CIMAP has now been accredited in the areas of crop production, processing and trading of medicinal and

aromatic plants by ECOCERT in accordance with requirements of National Organic Programme (NOP) US,

National Programme for Organic Production (NPOP) India, and the Commission Regulation (EEC) 2092/91 and

Regulation (EC) 1788/2001.

CIMAP has also taken an initiative on Development of Good Agricultural Practices (GAPs) in MAPs. The work has

been initiated with a move towards 'Green Agriculture' for developing practices requiring minimum essential

synthetic agri-chemicals for achieving higher yields, safer products with a clear objective of sustainability. Initial

studies conducted with Ocimum sanctum clearly indicated that chemical fertilizers can be reduced to an extent

of 70% when used along vermicompost. In this context, it has been established that perhaps, integrated

nutrient management is the most viable proposition.

IMPROVED VARIETIES & AGROTECHNOLOGIES

GENETIC IMPROVEMENT & BREEDING EFFORTS

Evaluation of performance of a linalool-rich variant of Ocimum basilicum

Characterization of Ocimum germplasm lines

Screening and evaluation of Claviceps purpurea for improving productivity of ergot and ergot alkaloids

A high yielding genotype of lemongrass (Cymbopogon khasianus) 'NS-1' suitable for marginal lands

Morphological tag developed in low latex line of poppy (Papaver sommiferum) for seed crop

Identification of a high yielding strain CIMAP–SR 3-3 of Stevia (Stevia rebaudiana) for northern Indian plains

Development of a high yielding dark purple pigmented strain STAS-3 of Shyam tulsi (Ocimum sanctum)

Four chemovariants of Ocimum basilicum clones rich in linalool were identified from a population raised from

open-pollinated seeds. The contents of linalool in these variants varied from 49-73%. The variant with highest

content of linalool i.e., 73% was multiplied and its seeds were produced in isolation through intermating of

plants raised from stem cuttings. The resulting two generations were evaluated during two years at four plant

growth stages. Herb yield, oil content and linalool content increased from pre-emergence to seed-set stage.

Further, no deterioration in linalool content was noticed over two generations of evaluation when seeds were

produced through open-pollination in isolation.

The Ocimum germplasm bank at CRC, Hyderabad contains 39 germplasm lines comprising of 4 species i.e. O.

basilicum, O. tenuiflorum, O. gratissimum and O. kilimandscharicum. Studies for morphological, chemical and

molecular characterization were initiated at CRC, Hyderabad during 2007-08 in order to assign descriptive

characters to the genotypes for precise identification. Morpho-agronomical characterization (52 traits) of the

39 Ocimum genotypes has been done.

Field study on yield and quality evaluation of ergot was conducted. Maximum calculated yield was more than

150kg/ha in five accessions while in remaining accessions (except two where it was less than 100kg /ha ),yield

was more than 100kg/ha.This yield performance was only upto mid November sown crops. Beyond that

sclerotial yield was dastically reduced upto12kg/ha in one of the accessions.

Two superior genotypes of lemongrass were identified for vigour, high herb and oil yield with high citral

content. Genotype NS-1 was found to be highly promising for very fast growth, high herb and oil yield of better

quality.

A recombinant genotype of opium poppy has been developed in near zero latex containing variety Sujata

background to distinguish its plants from other latex producing varieties of poppy. The new recombinant has

the distinguished marker in form of deep fringed leaves.

Two superior genotypes SR- 2-6 and SR 3-3 were identified where one genetic stock, namely SR 3-3 was found

to be highly promising for high fresh and dry leaf yield with high steveoside, rebaudioside with low dulcoside - A

content (%) in the leaf.

Under genetic improvement programme of tulsi (Ocimum sanctum), the diverse genetic stocks of Shyam tulsi

(the purple pigmented tulsi plant) were collected from Uttar Pradesh, Bihar, Madhya Pradesh, Orissa,

Uttarakhand, Gujarat, Maharashtra, Jammu & Kashmir, West Bengal states of India. The strain STAS - 3

consistently showed high oil content in BST and PST.

2 3

Annual Report 2008 : Highlights Central Institute of Medicinal and Aromatic Plants

PROCESS TECHNOLOGIES, VALUE ADDITION AND UPSCALLING

Upscaling of improved processing technology for isolation of hepatoprotective CLIV 92 from Cleome viscosa

Upscaling and improvement of extraction technology for isolation of calliterpenone from Callicarpa

macrophylla

Improved process for the preparation of antimalarial drug artelinate (AL)

Improved process technology for dihydroartemisinin

Aloe vera processing technology

Post harvest storage method for rose-scented geranium (Pelargonium graveolens) herb under hill conditions

of Uttarakhand

A novel processing technology was developed for isolation of Cliv-92 from the seeds of Cleome viscosa. The

technology has now been upscaled to a level of 50 kg / batch size on the pilot plant. The novel processing

technology is based on the use of solid matrix adsorption method followed by partitioning with different

organic solvents.

The process technology developed for the isolation of the molecules has now been upscaled to a level of 30 kg /

batch. Optimisation of the process parameters like extracting solvent, temperature of extraction, and

downstream techniques have been optimized, resulting in many fold increase in the yield of the product. One

of the major improvements in the upscaled technology is the application of membrane filtration technique for

the concentration of the extract and removal of water.

Two improved processes for the preparation of artelinate have been developed. In the first process reduction,

alkylation and hydrolysis steps were carried out in one pot followed by work up and recrytallization to produce

pure AL with 81-88% yield in 10-12 hours. In the improved (second) process, removal of unwanted, undissolved

reaction produced after completion of reduction and alkylation steps improved the hydroysis step and produce

much cleaner product (AL) which upon direct recrystallization, discarding the use of column chromatography,

produced pure AL with 95-98% yield.

An improved process technology and know-how for the conversion of artemisinin to DHA has been developed.

It reduces process steps of distillation of solvents, extraction, recrystallization, thus making the process

economic, greener, less time consuming and having higher overall yield of DHA, 95% w/w, with 97 % purity.

CIMAP has developed an improved and economical technology with better process efficiency and superior

quality for the processing of Aloe vera for aloe juice, filleted leaf sap, Aloe vera gel and Aloe vera spray dried

powder.

Geranium biomass can be stored in polythene bags up to 48 hrs without losing the essential oil. It was observed

that esters (formate, acetate, butyrate and tiglate of citronellol and geraniol), neral and 10-epi--eudesmol

improved and ketones (menthone, isomenthone) and alcohols (linalool, nerol, geraniol) decreased on post

harvest storage. It is also noted that citronellol to geraniol ratio (C: G) improved on storage in the polythene

bags.

Post harvest storage effect on essential content and composition of Salvia sclarea grown in Kumaon region

of western Himalaya

High-performance thin-layer chromatographic method for quantitative analysis in Clerodendrum inerme

BRP-HPLC method for the quantitation of glabridin in “Yashti-madhu” (Glycyrrhiza glabra)

NMR method for small molecule structural studies

Quality-Marker development in Gloriosa superba and Coleus forskohlii grown in South India

Adulterant of commercial 'Guggul' identified

Method for identification and quantification of polyisoprenylated benzophenones of Garcinia cambogia

Storage of clarysage inflorescence under shade for 96 hours prior to distillation significantly reduced the

essential oil recovery (from 0.30 –0.13%). Linalool was found to increase on storage (28.86% to 37.47%), while

linalyl acetate initially increased (from 39.68% to 43.30%) otherwise it decreased on storage.

A sensitive, selective, precise, and robust high-performance thin-layer chromatographic method has been

developed and validated for analysis of two new sterols, 4-methyl-24 ß -ethyl-5-cholesta-14,25-dien-3 ß -ol

and 24 ß -ethylcholesta-5,9(11),22E-trien-3 ß -ol, and a triterpene, betulinic acid, in Clerodendrum inerme

extract.

A reverse phase high performance liquid chromatographic method has been developed for the quantitative

estimation of glabridin in Glycyrrhiza glabra, using C18 column with acetonitrile-water containing 2% AcOH

(70:30) as an eluent.

To avoid unnecessary time delays and at the same time to achieve better sensitivity, DEPT-Q type experiment

was standardized and brought into routine use. A typical DEPT-Q spectrum reveals all four types of carbon

resonances with primary and tertiary on one phase and rest on the other phase.

Extraction of the marker compounds from the herb Coleus forskohlii was done at 2 kg level. The marker

compound, Forskolin, was isolated in pure form (>95% purity by HPLC) from Coleus forskohlii roots by column

chromatography and used as reference standard. Structure confirmation of the compound was done by NMR

spectral studies. HPLC Quality control method was developed and standardized for the analysis of forskolin.

Gum-oleo-resin of guggul is usually adulterated with the allied species C. agallocha, which is medicinally not of

much importance. Both the species and the commercial samples of guggul were pharmacognostically

evaluated and diagnostic characters were identified in the present study to distinguish the authentic sample

from its adulterant.

A sensitive liquid chromatography/electrospray ionization tandem mass spectrometrical (LC/ESI-MS/MS)

method was developed for simultaneous identification and quantification of two polyisoprenylated

benzophenones, isoxanthochymol and camboginol in the extracts of the fruit rinds, stem bark,seed and leaves

of Garcinia indica and in the fruit rinds of Garcinia cambogia.

NEW ANALYTICAL METHODS

4 5

Central Institute of Medicinal and Aromatic Plants Annual Report 2008 : Highlights

plantlet production and herb yield and incidence of black spot disease on Aloe vera.Maximum plant height

attained was 56 cm in Glomus intraradices treated plants followed by 53cm in Bacillus subtilis and B. subtilis +

Glomus aggregatum treatments. Maximum plantlets production (14) was recorded in the treatments of B.

subtilis + Trichoderma harzianum + G. aggregatum followed by 12 in others. Maximum herb yield obtained was

3.65 kg/plant in G. aggregatum treated pots, followed by 3.35 kg/plant in G. aggregatum + B. subtilis and

Glomus intraradices treated pots.

Six applications of mancozeb @ 0.3% as foliar spray were found to be most effective giving 187.8 g fresh

root/plant and 78.8% protection against foliar blight/spot (A. alternata, C. rauvolfia, C. serpentina,

C. gloeosporioides, C. cassiicola, C. lunata, M. phaseolina and R. solani) diseases. Application of two doses of

neemcake @ 50 g/plant at 30 days interval plus two sprays of B. subtilis in September and October along with

three foliar spray of mancozeb at 15 days interval plus two foliar sprays of carbendazim @ 0.15% at 21 days

interval provided, 201.5 g fresh root/plant, and 75.5% protection against foliar blight pathogens.

Nine chemical activators namely, isonicotinamide, 2-chloronicotinic acid, 5-nitrosalicylic acid, 4-chlorosalcylic

acid, DL-2 aminobutyric acid, 2-aminobutyric acid, O-acetylsalicylic acid, 4-aminosalicylic acid and salicylic acid

were used as soil drench on three week old seedlings which were planted in root-knot nematode infested soil.

Maximum reduction in root-knot severity and nematode population occurred with 4-chlorosalycylic acid, O-

acetylsalicylic acid and 2-chloronicotinic acid. The result envisages that these chemical activators may be used

in integrated management system for protection against root-knot nematodes.

The experiment conducted on integrated pest-management in mints indicated that initial adult population of

white fly was found building up from March and was maximum in the month of May. Among genotypes

evaluated for this resistance potential to whitefly population, significantly low number of flies observed in

variety - Kushal. Out of four samplings in field from March to May, the Begomovirus infection was also found

minimum in variety - Kushal as compared to others. The maximum infestation of Begomovirus was found in

cultivar Shivalik. The genotypes; MAS-35, MAS-216 and cultivar Shivalik were found moderately resistant to

gall forming root - knot nematode (Meloidogyne incognita). However, none of the varieties/genotypes was

found to be highly resistant or susceptible with respect to outbreak of M. incognita.

Highest root biomass (15.20 q/ha) and oil yield (40.10 kg/ha) was recorded in vetiver when harvested at age of

10-12 month followed by 9 month aged crop. The vetiver crop as annual crop successfully be grown under the

sub-tropical climate of Uttar Pradesh for better economic returns.

Two distinct leaf blight diseases of kalmegh have been investigated. One type is caused by Colletotrichum sp.

and another by Rhizoctonia sp. Rhizoctonia was found more destructive than the Colletotrichum sp. However,

both caused premature defoliation of leaves. In Senna the disease was found to be caused by Colletotrichum

Integrated management of foliar diseases of sarpagandha in field

Chemical activators: A novel and sustainable approach for management of Meloidogyne incognita in

Chamomilla recutita

Integrated pest management in mints

Effect of crop age on root and oil yield of vetiver

Leaf blight disease of Kalmegh (Andrographis paniculata), Senna (Cassia senna) and Aloe vera

Separation and quantification of lignans in Phyllanthus species by a simple chiral densitometric method

Influence of sources and doses of N and K on herbage, oil yield and nutrient uptake of patchouli in semi-arid

tropical climate

Influence of integrated nutrient management on yields, oil quality and soil fertility of rosemary

Qualitative performance of Java citronella (Cymbopogon winterianus) cultivars in Uttarakhand hills

Influence of different VAM fungi on the root biomass yield and reserpine content of Sarpgandha (Rauvolfia

serpentina) in unsterile soil

Effect of date of planting on the yield and disease incidence of Aloe vera

Influence of bioinoculants on the growth, yield and black spot disease of Aloe vera

A sensitive, selective, and robust high-performance TLC (HPTLC) method using chiral TLC plates for qualitative

and quantitative analysis of phyllanthin, hypophyllanthin, niranthin, and nirtetralin, the active lignans of

Phyllanthus species, was developed and validated.

Field experiments were conducted at CIMAP RC, Bangalore to study the influence of sources and doses of N and

K on herbage, oil yield, nutrient uptake, nitrogen use efficiency (NUE) and oil quality of patchouli and soil fertility -1 -1status. The results revealed that application of 200 kg N ha and 41.5 kg K ha produced significantly higher

patchouli herbage and oil yields compared with control.

Results revealed that all the treatment combinations significantly influenced the herb and essential oil yield over

control. Herb yield increased by 39-85% over control with application of different combinations of inorganic

fertilizers and organic manure (vermicompost). The corresponding increase with full nutrient supply through

the organic manure and full nutrient supply through inorganic fertilizers was 39 and 85%, respectively. Similarly,

oil yield increased by 37-84% over control with different combinations.

Hydrodistilled essential oils of Java citronella (Cymbopogon winterianus) cultivars grown in Uttarakhand hills

were analysed during summer, rainy and winter seasons. Cultivars Manjusha, Mandakini, Jalpallavi and Bio-13

recorded highest oil in winter (1.5 –1.82%), while cultivar Medini (1.1%) in summer season. In cultivar Manjusha,

Mandakini and Bio-13, citronellal content was higher in summer followed by rainy season, while cultivar

Jalpallavi recorded maximum citronellal content during rainy season. Cultivar Medini was geraniol rich, and

geraniol was found to be higher during summer (45.74%) followed by winter (43.71%).

The sole treatment of G. mosseae, G. fasciculatum, G. margarita and G. aggrigatum increased the fresh root

biomass yield of sarpgandha over untreated control by 203, 157, 135 and 103%, respectively.

Maximum herb yield/ plant (3.5kg) of Aloe vera was recorded in March to May planting with 15-17 months

duration. Yield was recorded to be more than 3kg/plant in 12-14 month- old plants. Thus, these plants can be

very safely cultivated as annual without any significant effect on herb yield.

Four selected bioinoculants namely, Bacillus subtilis, Glomus aggregatum, G. intraradices and Trichoderma

harzianum were used alone and in different combinations to evaluate thier influence on plant height, number of

INTEGRATED NUTRIENT AND PEST MANAGEMENT

6 7


grow in presence of 25 ppm of Cr in soil, became stunted and died with higher accumulation of Cr in root (989

ppm) and leaves (415ppm) as compared to that grown in normal soil taken as control.

The herb and oil yield of M. piperita significantly increased with the application of heavy metals to soils up to

certain level. The relatively high concentration of Cr, Cd, Pb and Ni in root tissues than in the shoot tissues

indicate that the M. piperita restricted the translocation of heavy metal from root to shoot. These results

indicate that the M. piperita could be grown on moderately heavy metal contaminated soils.

The study indicated that vermicompost made up of distillation waste of aromatic grasses besides other

beneficial effect on soil physical, chemical and biological properties, also helps in regulating the availability of

trace elements in soil.

Active constituents curcumin, demethoxycurcumin and bisdemethoxycurcumin were isolated from the

rhizomes of C. longa. The chloroform extract showed significant antimycobacterial activity. On isolating the

pure molecules based on activity guided fractionation, demethoxycurcumin was found to be the active

constituent. Some of its derivatives were also synthesized for structure and activity relationship studies.

Antimicrobial and drug resistance modifying activity of the essential oil from Rosmarinus officinalis was

evaluated against human pathogenic bacterial strains using disc diffusion and micro broth dilution assay. The

oil was found to be more active against the gram positive human pathogenic bacteria compared to gram

negative bacteria used in this study. The present finding suggests that characterization and isolation of the

active compound(s) from the rosemary oil may be useful in the treatment of bacterial including drug resistant

infections.

As a part of efforts to discover plant based biologically active molecules/compounds through bioprospection,

362 extracts and essential oils from MAPs were evaluated for their antimicrobial activity. Out of 311 extracts,

146 were found active against bacteria, 82 against fungi and 37 against both (bacteria and fungi). Out of 51

essential oils, 37 were found active against bacteria, 50 against fungi and 33 against both.

Effect of seasonal variation on biological activity of O. biennis root extract was studied against Staphylococcus

aureus and S. epidermidis. The roots were collected between April 2006 to July 2006 at 15 days interval and

extracts prepared were subjected to activity evaluation both qualitatively and quantitatively. The maximum

activity was observed in the samples collected in the month of June - July.

Effects of heavy metals (Chromium, Cadmium, Lead and Nickel) on the growth and yield of Mentha piperita

Vermicompost as a carrier as well as organic chelate for regulating Fe and Zn availability in soil and growth

and yield of geranium (Pelargonium graveolens)

Demethoxycurcumin: An antimycobacterial principle from turmeric (Curcuma longa)

Rosemary oil useful against drug resistant bacteria

Bioprospecting for antimicrobials from plant sources

Effect of seasonal variation on antibacterial activity of Oenothera biennis roots

BIO ACTIVITY AND DRUG MOLECULE PROSPECTION

sp.(new species), which leads to premature defoliation of the leaves. This is one of the serious diseases of Aloe

vera, caused by Colletotrichum sp.; the disease greatly reduced the yield of gel. The disease have been

successfully managed by three application of Tilt (Propiconazole) @ 0.1% at 15 days interval.

The germination percentage in T. minuta accessions ranged from 17.3% to 28.6% depending upon their

genotypic differences. Similar trend in result was recorded for germination energy, the parameter to measure ththe speed of germination. The germination period was mostly 10 day in all the accessions.

The results of an experiment on intercropping of Mentha arvensis and vetiver indicated that 80 kg oil of M. 2

arvensis was obtained in one harvest from an area of 6000 m . The digging of vetiver root was done in the month

of the Jan 2007. Approximately 12 .0 kg oil of vetiver was obtained from the same area. Thus both the crops

planted in the month of March and Mentha arvensis harvested in the month of May produced 135 kg oil /ha,

and 20 kg oil /ha of vetiver. This intercropping of M. arvensis with vetiver, generates additional revenue and

helps in better utilization of resources and additional profits to farmers.

With a view to identify MAPs suitable for understorey cropping with Phal van (fruit orchard), four experiments

have been established: under artificial shade, under Anola orchard, under Eucalyptus citrodora and Bananna

plantation and under natural shade of mango orchard at Central Institute for Sub-tropical Horticulture,

Lucknow. The experiments indicate that many crops like Kalmegh, Asparagus Patchouli etc. are moderate to

highly shade tolerant.

The herb and oil yield of Japanese mint significantly increased with the application of all the heavy metals to

soils to a certain level. The increase in the yield was 20.9, 35.2, and 34.2 % in herb and 27.5, 13.7, and 6.8 % in oil

with the application of 15, 30 and 45 mg Cr / kg soil, respectively. The menthol content in the essential oil was

not significantly affected by the application of heavy metals to the soil.

An experiment was conducted to study the effect of urease, amylase, dehydrogenase, acid phosphatase and

alkaline phosphatase on geranium, rosemary and Artemisia in rhizosphere. The different distances from the

mesh surface (0-15, 15-30, 30-45 cm) were analyzed for activities of different enzymes involved in C, N, P and S

cycling. The enzyme activities showed a clear gradient up to 0-15 cm depth for the root surface, while no such

gradient were observed in the unplanted soil. Enzyme activities were higher in the geranium and rosemary

rhizosphere, whereas in Artemisia rhizosphere the activity was found to be low.

The results of the experiment to assess the influence of chromium on growth and yield of kalmegh and the

ability of organic matter to mitigate the toxic influence of chromium has revealed that Kalmegh was unable to

Comparative germination behaviour of marigold (Tagetes spp)

Intercropping of Mentha arvensis with vetiver for higher returns

Exploring companion cropping of MAPs in agroforestry and fruit orchards by shade tolerance evaluation

Japanese mint as potent crop for the effective utilization of heavy metal polluted soils

Variability of soil enzyme activities in the rhizosphere of geranium, rosemary, Artemisia and unplanted crop

Organic matter as amendment to mitigate the toxic influence of chromium on kalmegh

PHYTOREMEDIATION AND SOIL QUALITY MANAGEMENT

8 9


Antimicrobial potential of Glycyrrhiza glabra roots

Synergism between extract of Phyllanthus amarus and silymarin for liver protection

Isolation of bioactive phytomolecules from Bidens pilosa

Bioactive phytochemical leads as mosquitocidal agents

Gallic acid based steroidal chalcones as anticancer agents

Anticancer and hepatoprotective agent from Withania somnifera and biological evaluation of its ester

derivatives

Cytotoxic agents from Terminalia arjuna

Antimycobacterial activity of Glycyrrhiza glabra was found at 500 µg/mL concentration. Bioactivity guided

phytochemical analysis identified glabridin as potentially active against both Mycobacterium tuberculosis H Ra 37

and H Rv strains at 29.16 µg/mL concentration. It also exhibited antimicrobial activity against both Gram-37

positive and Gram-negative bacteria. These results indicate potential use of licorice as antitubercular agent.

In search of the effective and standardized hepatoprotective combination therapy, silymarin and standardized

extract of Phyllanthus amarus were evaluated against CCl induced hepatotoxicity in rats. The combination of 4

silymarin and Phyllanthus amarus exhibited synergistic effect for hepatoprotection and silymarin with

ethanolic extract of P. amarus showed better activity due to the higher concentration of phyllanthin in ethanolic

extract in comparison to aqueous extract of P. amarus as estimated by HPLC.

The phytochemical investigation of hexane extract of Bidens pilosa leaf gave a bright orange compound ß-

carotene, phytyl alkanoate, friedelin, stigmasterol, 1-phenyl heptatriyne and a carotenoid derivative.

Compound 4'-O-â-D-(4”,6”-diacetyl)-glucopyranoside showed significant antioxidant activity and was found to

be non toxic to erythrocytes in in vitro bioassays.

Essential oil of blue stem grass, Bothriocola bladhii with sirodecenone and camphene as the major constituents

was observed to exhibit bioactivities against mosquito species, Aedes aegypti. Essential oil was evaluated for

ovicidal, larvicidal, growth disruption and adulticidal activities. Out of various bioactivities evaluated, the oil

was found effective only as larvicidal.

Estradiol based steroidal chalcones were synthesized. Using MTT assay, the anticancer activity of all these

synthesized chalcones was evaluated against various human cancer cell lines. Several steroidal chalcones

showed higher levels of cytotoxicity.

Detailed chemical investigation of Withania somnifera roots resulted in the isolation and identification of a

cytotoxic and hepatoprotective agent, palmitic acid, which was converted to eight semi-synthetic ester

derivatives. 2,9 t-Butyl palmitate and amyl palmitate were 4-6 times more active than against adherent and

suspension colon cancer cell lines. Interestingly, palmitic acid and its ester derivatives also showed

hepatoprotective activity which is being reported for the first time.

Four novel cytotoxic agents arjunic acid, arjungenin, arjunetin and arjunoglucoside-I were isolated from the

bark of T. arjuna. Out of the four compounds, arjunic acid was significantly active against the human oral (KB),

ovarian (PA 1) and liver (HepG-2 & WRL-68) cancer cell lines. Further, the most active compound arjunic acid

was converted into seven semi-synthetic ester derivatives 5 - 11. 2-O-Palmitoyl arjunic acid showed two times

more activity, while 2, 3-di-O-acetyl-, 2-O-p-anisoyl-, 2, 3-di-O-benzoyl- and 2, 3-di-O-p-nitrobenzoyl arjunic

acid showed 1.7 - 2.3 times less activity than the cytotoxic drug vinblastine against the liver cancer cell lines

HepG-2 and WRL-68, respectively.

The methanol extract of the fruit rinds of Garcinia indica showed potent cytotoxic activity against three human

cancer cell lines- colon (COLO-320-DM), breast (MCF-7) and liver (WRL-68) as determined by the MTT assay.

Through in-vitro bioassay, 160 plant extracts from CIMAP repository were evaluated against human; 4 plant

extracts showed the cell line dependent cytotoxicity in terms of IC-50 and IC-90 values against all the human

cancer cell line used. Further of interest, these extracts did not exhibit activity against the mice hepatocytes up

to concentration of 100 µg/ml.

Under Control and Supervision of Experimentation Animals (CPCSEA) all institutions carrying animal

experimentation have to constitute Institutional Animal Ethics Committee (IAEC) and animal experiments have

to be approved by the CPCSEA. The protocols have been categorized into infecting disease, metabolic

disorders, and pharmacological and toxicity studies. We have tested a total 259 smaples out of which 73 were

found positive.

O. vulgare collected from the high altitude of Himachal Pradesh (Kothi-2500m) was found to be a new

chemotype rich in thymol (62%), p-cymene (9%) and 3-octanone (8%). Highest thymol content of 49% has so

far been reported in the oil obtained from plants of Mediterranean region .

The essential oil obtained from the inflorescence of Artemisia capillaris was analysed by GC and GC-MS. Forty

two compounds representing 93.82% of the oil were identified. The main constituents were capillene,

terpinene, eugenol, limonene, p-cymene, β-myrcene and -pinene.

Volatile fractions from flowers of four geranium chemotypes viz., Bourbon, CIM-Pawan, Kelkar and Algerian

were analyzed by capillary GC. Thirty compounds comprising 91.67%of the Bourbon, 29 compounds

representing 93.07% of the CIM-Pawan, 23 compounds representing 85.37% of the Kelkar and 26 compounds

accounting for 87.48% of Algerian flower oils have been characterized. Bourbon and CIM-Pawan were found to

be rich in citronellol (25.64% and 28.44%), geraniol (25.21% and 23.85%) and citronellyl formate (10.11% and

11.18%). Cultivar Kelkar had citronellol (62.48%) and isomenthone (12.67%) as principle components.

However, Algerian type was found to be rich in citronellol (29.92%), citronellyl formate (23.22 and %) and

geraniol (7.21%).

Cytotoxic activity of Gracinia indica

Cancer cell line dependent cytotoxic activity of essential oil as compared to pure compound

Pre-clinical study at “Jeevanika” (Animal house)

A new chemotype of Origanum vulgare identified

Essential oil constituents of Artemisia capillaris from Uttarakhand hills

Essential oil composition of flowers of geranium (Pelargonium graveolens) chemotypes from Uttarakhand

hill

PHYTOCHEMICAL PROSPECTION

β

10 11


Cloning of norcoclaurine synthase (ncs) gene involved in isoquinoline alkaloid biosynthesis in poppy

(Papaver somniferum)

An alpha-1 antitrypsin genetic variant identified in human liver biopsy sample

Polymorphism study in different chemotypes of ashwagandha (Withania somnifera) through AFLP

Semi-quantitative RT-PCR analysis of leaf-specific gene dat in Catharanthus roseus cv. “Dhawal”

Cloning of genes involved in isoquinoline alkaloid biosynthesis in poppy

Novel cytochrome P450 monooxygenase genes (cyp) from Artemisia annua

Semi-quantitative RT-PCR analysis of seasonal variation in transcript abundance of leaf specific/prominent

genes dat and sgd in Catharanthus roseus cv. “Dhawal”

The gene for this early pathway step in morphinan alkaloid biosynthesis has been isolated, cloned and

sequence-characterized from Papaver somniferum. This will prove beneficial for further metabolic engineering

of the morphinan alkaloid biosynthetic pathway in heterologous systems.

The study was carried out as an initial step for characterizing the AAT gene in an Indian sample with the ultimate

aim of developing a heterologous alpha-1 antitrypsin production system. The AAT gene was isolated and cloned

from a liver biopsy sample through RT-PCR and the full-length gene was sequenced.

AFLP analysis of 20 different W. somnifera germplasm accessions representing various chemotypes (for

withanolide content), present in the National Gene Bank of Medicinal and Aromatic Plants (NGBMAP) at

CIMAP, was carried out. A total of eight primer pairs (EcoRI/MseI) were used for the analysis, which yielded a

total of 4821 bands of which 4701 were polymorphic and 336 were unique thereby indicating 95.13 %

polymorphism among the chemotypes. This study will help in classifying the Withania germplasm for targeted

genomics studies in future.

Semi-quantitative RT-PCR analysis was carried out to measure the transcript abundance of leaf-specific gene

dat in the aerial and underground parts of the C. roseus (cv “Dhawal”) plant at three different plant

developmental stages (6 days, 6 weeks and 6 months). This gene is significant since its protein product catalyzes

the last step of vindoline biosynthesis, which then commits the metabolic flux into bisindole alkaloid

biosynthesis. Transcript abundance of the dat gene was found to decrease with the plant age.

Four new genes have been cloned from opium poppy cv “Sampada”. These genes are S-adenosyl-L-

methionine:norcoclaurine 6-O-methyltransferase (6omt) and S-adenosyl-L-methionine:3'-hydroxy-N-

methylcoclaurine 4'-O-methyltransferase (4'omt) in the pre-reticuline pathway and salutaridinol 7-O-

acetyltransferase (salAT) codeinone reductase (cor) in the post-reticuline pathway.

A bioinformatics-based approach has been used to hunt for novel cyp genes from Artemisia annua, whereby

the information from the conserved domains of cytochrome P450 genes have been utilized for cyp gene

amplification. Using primers based on these conserved domains, partial sequences of 22 novel cyp genes were

obtained, of which, 2 (cyp73 and cyp83) have been isolated full length using 5'-RACE.

The dat transcript levels in the mature leaf of 6 months old-field grown plant were found to be higher in winter

(December) than in summer (June) season. The sgd transcript levels in the mature leaf of 6 months old-field

grown plant did not vary much with the seasonal change but were found to be slightly higher in summer (June)

than in winter (December) season.

Variation in the chemical composition and yield of essential oil of rose scented geranium (Pelargonium

species) by foliar application of metallic salts

New thaiocarbamates from Moringa oleifera

Composition of North Indian pyrethrum (Chrysanthemum cinerariaefolium) flower oil

Effect of altitude on the essential oil constituents of a new variety of Artemisia (A. roxburghiana var.

purpurascens)

Heterologous expression of menthol biosynthetic pathway gene in bacteria

Cloning of cyp71AV1 gene from Artemisia annua (variety “CIM Arogya”)

The concentration of linalool, geraniol and 10-epi- gamma eudesmol in the essential oil of geranium

significantly increased by the application of metallic salt, whereas the concentration of isomenthone,

citronellol and citronellyl formate was not significantly affected as compared with control plants. The oil yield

of geranium significantly increased by the foliar application of metallic salts as compared with control plants.

The increase in the oil yield was 41.9, 23.4, 29.6, 29.6, 41.9 and 41.9 % over control by the application of sodium

chloride, calcium chloride, iron sulphate, zinc sulphate, copper sulphate and nickel sulphate, respectively.

From the pods of Moringa two new thiocarbamates, O-n-butyl 4-[(á-L-rhamnopyranosyloxy) benzyl]

thiocarbamate (E) and O-ethyl-4-[(á-L-rhamnopyranosyloxy)-3hydroxybenzyl] thiocarbamate (E) along with

niazirin and 5,5-dimethyloxazlidine-2-thione were isolated.

Twenty two compounds representing 85.27% of the total oil were identified. Trans-chrysanthemumic acid

(19.61%), decanoic acid (7.63%), nerolidol (9.63%), spathulenol (4.58%), guaiol (5.93%), â-eudesmol (9.81%)

and á-eudesmol (7.07%) were the major compounds. The oil of Indian species is uniquely different from the oil

obtained from pyrethrum of Kenya.

In-vitro testing of dried ethanolic extracts of the plant showed promising insulin secretagogue activity. Plants

collected from different altitudes (850m, 1218m and 2205m) of Garhwal Himalayas showed considerable

variation in yield and quality of oil. The oil yield was lowest (0.2%) in the plants collected from higher altitudes

and it was rich in borneol (21.2%) followed by linalyl acetate (7.4%) and á-humulene (6.7%). While, the plants

collected from the lower altitudes yielded higher percentage of oils (0.8-0.85%) which were dominated by â-

caryophylene (16.3-18.4%) followed by eugenol (16.2%)or á-thujone (12.0%).

The menthol biosynthetic pathway gene pulegone reductase (pr) cloned and sequenced earlier at CIMAP

(submitted to NCBI under Accession Number EF426467) from Mentha arvensis was cloned in pET100 vector

and expressed in bacterial E. coli strain BL21 (DE3) background.

The gene 1467bp for cyp 71AV1, which catalyzes the three step oxidation of amorpha-4,11-diene to artemisinic

acid in the artemisinin biosynthetic pathway of A. annua has been isolated, cloned and sequence-

characterized.

PLANT GENOMICS & MOLECULAR BIOLOGY

12 13


14in shoot cultures was corroborated with radiolabel incorporation studies using [2- C] acetate as precursor.

Production of withaferin A was also found in the in vitro shoot cultures.

Method for hairy root induction was developed in Solanum nigrum. In vitro raised plant's leaf and stem were

used as explant source for the Agrobacterium mediated genetic transformation of Solanum nigrum. Infected

plant tissues after 2-3 weeks of incubation showed the hairy root induction.

CIMAP has developed an Agrobacterium rhizogenes-mediated hairy root line (CR-N2) of C. roseus (Var.Nirmal)

that exhibits unique capability to regenerate transgenic plants via adventive shoot bud organogenesis on a

modified medium.

The multiple shoot cultures of H. perforatum entered the exponential phase of in vitro growth between 15-25 th

days of the culture cycle (GI= 2379.29) followed by a gradual incremental biomass gain up-to 40 day (3173.69) th th

and a second rapid growth phase between 40 to 45 day of incubation (GI= 4328.07). Hypericine production thin these shoots occurred after 25 day of growth and reached the peak between 35-45 days of the culture cycle

that coincides with the onset of stationary phase of growth after the two exponential growth peaks.

Using young offset as explants, a rapid in vitro micro-cloning method has been developed to produce disease-

free stocks of Aloe vera (Var. CIM-Sheetal). A multiplication rate of one:eight was achieved within two weeks

when explants were cultured on a cytokinin containing medium.

Tissue culture studies in Chlorophytum borivilianum were undertaken for standardizing multiplication

protocols employing somatic embryogenesis aiming towards its high frequency rapid in vitro multiplication.

Growth kinetic studies were carried out in R. serpentina to find out a co-relation between growth period of the

root clones and higher biomass as well as secondary metabolite production. On the basis of behavior of the five

root clones it may be inferred that, alkaloid production in transformed roots in R. serpentina is associated with

growth and reserpine content was found to be highest at the end of active growth period i.e. 10 weeks of

culture duration.

Agrobacterium rhizogenes ( mediated hairy root cultures have been induced in leaf explants of

Rauvolfia serpentina at M

Establishment of hairy root cultures in Solanum nigrum

Plantlet regeneration from hairy roots of Catharanthus roseus

Biogenesis of hypericine in multiple shoots cultures of Hypericum perforatum as a function of culture age

and precursor feeding

Micro-cloning of Aloe vera

Induction and development of somatic embryos of Chlorophytum borivilianum in liquid culture medium

Growth kinetics studies in Agrobacterium rhizogenes mediated genetically transformed roots in Rauvolfia

serpentina

Establishment and chemical profiling of hairy root culture of Rauvolfia serpentina

strain A )4

81.25% relative transformation frequency. ost of the emerging roots of the present

study tend to form calli in the half-strength, hormone-free, liquid basal MS medium during further sub-

culturing.

Studies on drug resistance development and mechanism in Staphylococcus aureus through molecular

genetics approach

Role of efflux pump modulator in altering the activity of ATPase in Mycobacterium smegmatis

Bioconversion of a monoterpene into food flavoring agent vanillin

Standardization of cerebral malaria model in Mus musculus using markers of chemokine pathway

Modulation of inflammatory cytokines by Ocimum gratissimum under in-vitro conditions

Toxicity profile of Ocimum sanctum leaf at acute and sub-acute level in rat

Withanolide-A biosynthesis in in vitro shoot and root cultures of ashwagandha

In the present study, the sensitive Staphylococcus aureus (SA) and different drug-resistant mutants of SA

including MRSA were tested for their cross resistance to different antibiotics representing various structural and

functional groups. The ATPase activity of drug-resistant mutants were found to be decreased compared to

sensitive strain indicating less transport of drug thereby helps in acquiring resistance.

In presence of ciprofloxacin there was significant decrease in ATPase activity of drug-resistant mutant, when

compared with control suggesting effluxing of the drug in multi drug resistant mutant MDR-Q. However, in

presence of ciprofloxacin and plant molecule citral together, ATPase activity was increased.

The present work was done with the objective of the conversion of abundantly available phytomolecules

eugenol into vanillin a compound of industrial importance using microorganisms Aspergillus flavus, A. niger and

Pseudomonas aeruginosa. These microbes were found to be capable of converting eugenol to industrially

important cost-effective products, vanillin (used as flavouring agent).

In an effort to bioprospect medicinal plants for cerebral malaria a model was standardized for evaluating leads

against malaria using Plasmodium bergheii (Chloroquine sensitive) and P. yoelii nigeriensis (chloroquine

resistant) strains where in as a conventional parameter, parasitaemia and mean survival time (MST) are is being

monitored.

It was observed that O. gratissimum @ 1 and 10 mg/ml, exhibited the anti-inflammatory activity through

inhibition of pro-inflammatory cytokines (IL-1â and IL-6).No significant change were observed in IL-4

expression.

The in-vivo acute and sub-acute toxicity study of Ocimum sanctum (Aqueous extract) was conducted as per the

approved protocol by Institute's Animal Ethical Committee and confirmed to national guidelines on the care and

use of laboratory animals with an intention to study the safety profile of the plant (leaf) which is one of the most

commonly used medicinal plants in Ayurvedic system of medicine. It may be concluded that chemically

characterized aqueous extract of Ocimum sanctum leaf is well tolerated by Charles Foster rat at acute and sub

acute level.

Multiple shoot cultures of the experimental lines of Withania somnifera plants established using nodal segment

as ex-plant under specific hormonal combinations of benzyl adenine and kinetin resulted in significantly higher

level of biogeneration of withanolide A in the in vitro shoots. The enhanced de novo biogenesis of withanolide A

PLANT TISSUE CULTURE TECHNOLOGY

14 15


inhibitors involved and post-translational modification to its cloned commentary. Through user friendly GUI

based database one can find out all the relevant information about menthol and biosynthetic pathways.

DbAlbioPath is an alkaloid biosynthetic pathway database of medicinal and aromatic plants (MAPs). The

present database has graphical user interface (GUI). Through this user friendly GUI based database one can

find out relevant information about alkaloids and their pathways. The database contains information of the

alkaloids metabolic pathways and related genes. It comprises of chemical structures with name, formulae,

physical properties, SMILES generation, industrial application and map viewer of medicinal plants.

In this study, an in silico analysis was performed in which SSRs were mined from ESTs corresponding to alkaloid,

phenylpropanoid and terpenoid biosynthesis reported in a large number of important plants species .The

abundance and distribution of various type of SSRs were also analysed. Comparative genomic study was

performed for finding the corresponding genes to these SSR- ESTs related to alkaloids, phenylpropanoid and

terpenoid biosynthesis, using Arabidopsis thaliana as a model. Results revealed that the SSRs are distributed

throughout the complete datasets of EST, almost 941 trinucleotide, 213 dinucleotide, 157 tetranucleotide and

179 pentanucleotide reported in, Coptis japonica, Oryza sativa, Papaver somniferum, Catharanthus roseus,

Phaseolus vulgaris, Capsicum annuum, Mentha piperita, Euphorbia tirucalli. Primers associated with the SSR-

EST were also designed.

Cytochrome p450 protein sequences were analyzed to find a conserved sequence in several medicinal plants

(Mentha spicata, Ocimum basilicum, Stevia rebaudiana), and to find patterns involved in their active role. The

pattern analysis reflects motifs, which were found to be associated with the function of heme-binding activity

of Cytochrome p450 proteins, and were conserved in distant families of plant. The conserved motif sequences

were found to be involved in binding pocket formation, which validate their role in functionality of Cytochrome

p450.

Information pertaining to the different aspects of terpenes viz; classification of terpenes (monoterpenes,

diterpenes), physical properties of classified terpenes (for monoterpenes- linalol, nerol), categorization of

plants, chemical substances (toxaphene, geniposide), functional genes involved (cypla, ESR2) can be accessed

from this database.

Alkaloid biosynthetic pathway database (DbAlbioPath)

Bioinformatics based analysis of SSR's in ESTs corresponding to alkaloid, terpenoid and phenylpropanoid

biosynthesis

Pattern analysis in CYP450 proteins of some medicinal plants

Terpene bio-synthetic pathway database (Db-Terpene)

Chemical profiling of callus cultures of Taxus wallichiana

Rapid and efficient organogenesis in Mentha piperita

In silico insight into phytoremediation: Identification of metal binding motifs in medicinal plants

In silico identification of genes expressed in macrophage stage of Mycobacterium tuberculosis H37Rv strain

Menthol biosynthetic pathway database (MBPdb)

Five C-14 oxygenated taxoids were characterized from the analysis of the callus cultures of the Taxus

wallichiana which include the expression of

The

confirmation of the structures of the identified taxoids was made through their accurate molecular formula

determinations.

three previously identified taxoids, namely—yunnanxane (1),

2á,5á,10â,14â-tetraacetoxy-4(20),11-taxadiene (2) and 2á,5á,10â-triacetoxy-14â-(2-methyl)-

butyryloxy-4(20),11-taxadiene (3) and two newly identified taxoids, i.e.2á,5á,10â-triacetoxy-14â-

propionyloxy-4(20), 11-taxadiene(4) & 2á, 5á, 10â-triacetoxy-14â-isobutyryloxy-4(20),11-taxadiene (5).

A rapid and efficient method was developed to initiate multiple shoots from internodal and leaf explants.

Excised tissues (leaf blade and internode) were transferred in half strength MS medium supplemented with BAP

and TDZ. Multiple shoots were initiated within three-four weeks. Internodes / leaf blades with regenerated

multiple shoots were transferred to fresh half strength MS medium without supplements for elongation and

rooting.

An in silico analysis of metal binding proteins was performed for identifying putative metal binding proteins for

Cd, As, Hg, Co, Zn, Hg, As, Mo, Mn, Mg and Ni ions in medicinal plants. Related metal binding protein sequences

based on gene name were retrieved using UniProtKB/Swiss-Prot database. The conserved/consensus patterns

was obtained through MSA performed by ClustalX. Motifs were designed and validated through

UniProtKB/Swiss-Prot and UniProtKB/TrEMBL database search which yielded true positives showing the high

specificity of each motif.

Promoters with known transcriptional start sites for Mycobacterium tuberculosis genes in whole genome

(4,411,529 bp) was predicted, compiled and analyzed. Different nucleotide bases were detected as

transcription start site in different promoter sequences. In total of 1970 promoter sequences, adenine residue

was observed to be base for transcription initiation in 347 sequences, guanine in 608 sequences, thymine in 411

sequences and cytosine in 604 sequences. Thus the present study showed guanine was most frequently acted

as start site for the transcription. All the promoters were aligned to determine the consensus elements (-35

hexamer and -10 hexamer).

MBPdb database provides information of various enzymes participating in the pathway under different heads

e.g. EC number, pathway involved (related to KEGG database), reaction type, natural substrate, co-factors,

BIOINFORMATICS/ IN SILICO BIOLOGY

16 17


also known as plastid-pathway was very well conceived in all the above experiments conducted on

monoterpenes whereas the cytosol pathway oriented from MVA was found in the experiments conducted on

compounds of sesquiterpenoidal nature.

Artificial polyploids have been developed in a seed forming clone of vetiver. The polyploids thus developed

evince not only high gametic and seed sterilty but are also accompanied by enhanced biological potential for

the characters associated with increased cell size and physico-physiological efficiency. This has far reaching

implications in utilizing a given ideotype for efficient application of vetiver ecotechnology for its multifarious

environmental and industrial applications.

Analysis of alkaloid content showed that treatment duration of 12 hrs of Cr at 10 & 20mM and combination of

Cr- (10 & 20mM), Cd- (10 & 20 mM), and Mn- (10 & 20) with SA- (1 & 5mM) reduced alkaloid content possibly by

influencing biosynthetic steps. Treatments particularly Cd at (10 & 20 mM), Mn at (10 & 20mM) and YE at (1 & 5

g/L) and combination of Mn at 20 mM and 1 mM SA and YE at 5 g/L with SA at 5 mM showed enhancement in

total alkaloid content possibly by influencing biosynthetic pathway. These treatments can thus enhance

accumulation of total alkaloid in Catharanthus.

Results revealed that 0.00 Fe mg/l have the nil vinblastine and vincristine percentage with 0.93 % and 0.89 % of

total alkaloids in Dhawal-Ga treated and etheral treated plants. Vinblastine and vincristine are higher in Dhawal

etheral treted one then the Ga. Furthermore, the vincristine is totally absent in all the alkaloid of the 0.00 to

11.2 cultured plant of Catharanthus.

The virus was efficiently sap transmitted from naturally infected periwinkle plants to healthy plants and to a

variety of plant species mainly from Chenopodiaceae, Cucurbitaceae and Solanaceae. Total RNA was extracted

using Qiagen kit. The isolated RNA was used for cDNA synthesis using reverse transcriptase for PCR

amplification. The PCR reaction (25ìl) using primers specific to CMV coat protein (F:5'-

GCATTCTAGATGGACAAATCTGAATC-3'/ R:5'GCATGGTACCTCAAACTGGGAGCAC-3') was performed in an

automated thermal cycler. A part of the PCR amplification product was electrophoresed in 1.2% agarose gel in

1X TAE that exhibited an amplified DNA fragment of expected size (~650 bp) only from virus infected samples,

not from healthy ones.

About 179 accessions have been added to seed gene bank raising the present tally to accession of 2465 species.

In vitro bank has been enriched by 17 accessions and now comprises of 265 accessions. The DNA bank now

comprises of 1345 accessions with the addition of 23 accessions.

Towards development of non-invasive vetiver – Vetiveria zizaniodes

Elicitation studies in post-harvested leaves of Catharanthus roseus

Effect of gibberellin and ethereal and Fe nutrition in Catharanthus roseus tissue concentration and chemical

composition

Natural infection on periwinkle (Catharanthus roseus) by cucumber mosaic virus

GENE BANK STATUS/UPDATES

BASIC SCIENCE

Effect of zinc and paclobutrazol on growth and pyrethrin quantity and quality of pyrethrum

Effect of catechol treatment on post-harvested alkaloid content in leaves of Catharanthus

Changes in enzymes activities, proline, protein and IAA contents in relation to flower development in Rosa

damascena

Withanolide metabolism in ashwagandha (Withania somnifera)

Standardization of germination parameters in medicinal and aromatic plants (MAPs)

Mevalonate and non-mevalonate pathways for isoprenoid compounds in MAPs

Application of zinc enhanced pyrethric acid ester (Cinerin-2, pyrethrin-2 and jasmolin-2). Cinerin-2 -1

concentration increased significantly, and the increase was 53% and 40% over control at 5 and 10 kg ha of Zn

respectively, while pyrethrin-2 increased by 24% and 31%, respectively. Similarly, an increase of 10% and 42% in -1jasmolin-2 concentration was obtained by 5 and 10 kg ha zinc application, respectively.

The effects of catechol as elicitor for enhancement of alkaloid accumulation in post-harvested leaves of

Catharanthus roseus were investigated. The effect of catechol was dependent of dose and duration.

Treatments of catechol at 1.0mM at 12h resulted in elevation in content of total leaf alkaloid content.

Peroxidase, polyphenol oxidase, succinate dehydrogenase, amylase, catalase, proline content, protein content

and IAA content were studied at different stages. The higest peroxidase activity was recorded during green bud

stage and coloured bud stage coinciding with early flower development.

A high resolution DAD-ELSD-HPLC method of analysis of complex mixture of withanolides from leaves and roots

of Withania somnifera has been developed and validated through application of various chromatogram quality

parameters like peak resolution factor (R ), asymmetry factor (A ) and tailing factor (T ), RSD (%) of retention s s f

time and peak area response, LOQ, LOD, precision, recovery etc.

A study was conducted to elucidate the germination parameters: germination percent (%), germination energy

(%), germination period (days) and mean daily germination (%) in six major medicinal/aromatic plants: Tagetes

minuta, T.erecta, T. patula, Papaver somniferum, Plantago ovata and Vetiveria zizanioides. The seed weight

(test weight of 1000 seeds) in the six crops ranged from 0.447gm (in certain strain of T. minuta) to 1.784gm (in T.

erecta). It was interesting to record that in each of the six crops increase in test weight (seed size) almost linearly

enhanced the three seed germination parameters (germination percent, germination energy and mean daily

germination). The results led to the conclusion that seed size has greater role in controlling the germination

parameters in MAPs.

Studies were conducted to follow the biosynthetic pathway of different isoprenoids like menthol, menthone

and menthyl acetate (Mentha arvensis), artemisinin and artemisinic acid (Artemisia annua), patchouli alcohols

and hydrocarbons (Pogostemon cablin), citronellol, citronellal and geraniol (Cymbopogon winterianus), Citral-

trans and citral-cis (Cymbopogon flexuosus) etc. The most recent findings of non-mevalonate (DXP) pathway

18 19


PARTICIPATION IN AWARENESS EXHIBITIONS

TECHNOLOGY TRANSFER/ CONSULTANCY AGREEMENTS/CDA/ MoUS SIGNED

CIMAP participated in eleven exhibitions for showcasing CIMAP technologies, products, cultivars, seeds etc.

The visitors appreciated the R&D efforts of CIMAP in providing technologies to the farmers at the rural level and

to the entrepreneurs and industries at semi-urban and urban areas. Visitors took keen interest in CIMAP

products and purchased CIMAP publications, products, seeds and propagules. The visitors were supplied with

all information on agrotechnologies, processing, value addition and market potential of MAPs and were

encouraged to visit CIMAP and its Resource Centres and participate in the training programmes

conducted by CIMAP.

•Technology transfer on cultivation and distillation of patchouli (Pogostemon cablin Benth) to M/s Chennai

Bioflora Pvt. Ltd., Chennai.

•Technology transfer for production of Jasmine concrete and absolute to M/s. Sandana Florals, Golden

Jubilee Biotech Park for Woman society, Kanchipuram District (Tamil Nadu).

•Licensing of know how and design of improved field distillation unit and CIM-Asvika to M/s Swaraj Herbal

Plants Pvt. Ltd., Barabanki, Uttar Pradesh.

•Confidentiality agreement between CIMAP and EMAMI for pre-licensing evaluation of CIMAP products/

formulations: Myconil, HAloe Skin, Toothache lotion, Tooth Powder, Mosnobite, Mosaway, CIM-Paushak,

MosRep and, Herby Soft.

•Pre-licensing evaluation agreement with JK Agri-Genetics for complete technology package for novel plant

growth promoter “Calliterpenone” from Callicarpa macrophylla and Albizia lebbeck extract effective

against Bt-Cofton.

•Agreement with Biogentek BG (I) Pvt. Ltd. for exploring the possibilities of collaboration in the area of hairy

root culture/cell culture.

•Confidentiality agreement between CIMAP and Laila Nutraceuticals, Vijayawada for pre-licensing

evaluation of nutraceutical formulations and other leads of CIMAP.

•Confidentiality agreement signed with TVC SKY Shop, Mumbai for pre-licensing evaluation of CIMAP

herbal formulations.

•Memorandum of Understanding (MoU) for joint research project between CIMAP and University of

Mysore.

Technology licensing

Pre – licensing agreements

Collaborations

•Memorandum of Understanding (MoU) for joint research project between CIMAP and University of

Lucknow for studies on insect bioassay of plant extracts.

•Memorandum of Understanding (MoU) for joint research project with Ocimum Biosolutions, Hyderabad

on development of cost effective Microarray chip with periwinkle as a model.

•Agreement with Avitech Animal Health Pvt. Ltd., New Delhi for exploring the possibilities of collaboration

in the area of developing hepato-protective and immunostimulant product/feed for poultry.

•Designing, fabrication and development of plant/ herb distillation and processing units/ plants/

accessories for MAPs clients on demand at Narayan Mahavidyalaya, Goreakothi, Siwan (Bihar).

•Preparation of feasibility report for M/s SV Farms (P) Ltd., Hyderabad.

•Designing, fabrication and installation of two state- of- art distillation units for distillation of citronella and

lemongrass crops cultivated by the tribal farmers and supported by Integrated Tribal Development Agency

(ITDA), Govt. of Andhra Pradesh.

•Designing, fabrication and installation/commissioning of 1.0 ton capacity improved field distillation unit

on turn key basis to Mahamaya College of Agricultural Engineering and Technology, Akbarpur, Ambedkar

Nagar.

•Design/ Drawings for CIMAP improved FDU to Mr Vijayan, Vishakhapatnam.

•Survey of ash dyke area and preparation of a pre-feasibility report for NTPC, Unchahar, Raibareli.

•Survey of land and preparation of a pre-feasibility report for Vaid Organics, Hardoi.

•Crop consultancy in patchouli to Caps Ayur Krishi, Bangalore.

•Four CIMAP scientists visited foreign research establishments/institutions.

•Eighteen CIMAP scientists bagged reputed national awards/recognitions. CIMAP scientists were also

nominated to different academies, elected fellows and members of the high level expert/advisory

committees.

•Five Ph.D. degrees were awarded to the staff and students of CIMAP.

•More than 112 persons were trained at CIMAP, which included foreign scholars, students, industry

representatives and State and Central Government officials.

•First JNU-CIMAP Ph.D. Programme started.

nd •100 day CIMAP Certificate Course on Medicinal and Aromatic Plants (100 d CCC) held during 22 February thto 4 Junue, 2007.

CNP projects undertaken

HUMAN RESOURCE DEVELOPMENT

20 21


•CIMAP Summer Training (CST-2007) on Techniques and Tools of Biotechnology and Bioinformatics for th th

Graduate and Master Students held during 11 June-10 July, 2007.

•CIMAP Summer School (CSS-2007) on "Molecular Techniques in Bioprospection and Biodiversity th

Analysis" held during June 15th - July 30 , 2007.

•CIMAP Training School on Advance Instrumentation and Analytical Techniques for Natural Products (AIAT-rd th

2007) held during 3 -16 September, 2007.

th th•CSIR Programme for Youth for Leadership in Science (CPYLS) held during 15 -16 November, 2007.

•CIMAP Winter School (CWS-2007) on "Recent Techniques in Structural and Functional Genomics" st thheld during 1 -10 December, 2007.

•CIMAP Winter Training (CWT-2008) on “Techniques and Tools of Biotechnology and Bioinformatics for th thGraduate and Master Students” held during 20 January-20 February, 2008.

•A Unique "Know each other" meet of Scientists-Students and Teachers during CPYLS at CIMAP during 19th-

20th February, 2008.

th•National Technology Day (11 May 2007): BEE – MAPs 2007: Emphasis on creation of skilled human

resource: Farmers–industry–scientist converge at CIMAP; entrepreneurial training imparted to women

•'CIMAP Technology Fest-2007' Novel Community-Public-Private-Partnership model to show the way for th thrural prosperity (5 – 11 August, 2007) launched by Vice President, CSIR (Honorable Minister of Science

and Technology, Government of India, Shri Kapil Sibal). Aroma Biovillage Project jointly sponsored by DST

and CSIR, New Delhi was also launched.

th•CIMAP Day Function: MAPs show the way to adopt Integrative agriculture mode (Iam) (11 August, 2007)

•fgUnh lIrkg ¼14 flrEcj] 2007½ % lheSi esa fgUnh fnol lekjksg & tu&tu rd igWqpk;s fgUnh esa foKku

•lh-,l-vkbZ-vkj- LFkkiuk fnol % ̂gfjr [ksrh ls izkS|ksfxdh iFk&LoLFk lalkj dh vksj* ¼26 flrEcj] 2007½

th•MINT-2007: Interface on Mint (9 October, 2007) launched by Minister of State for commerce Sh. Jairam

Ramesh, Government of India, Jointly organized by CIMAP, Lucknow and Spice Board, Cochin

•jk"Vªh; oSKkfud laxks"Bh % ̂ ikni foKku }kjk LoLFk vkSj le`) lekt ds fy;s gfj;kyh dh [kkst* ¼29 uoEcj] 2007½% jktHkk"kk ds c<+rs dne

•fle&mRlo fdlku esyk 2008 ¼31 tuojh] 2008½

•National Science Day : Convergence of Science and Academia at CIMAP (28th February, 2008).

th •CIMAP Annual Day (26 March, 2008) : Exhibitions, field visit and lecture by eminent scientist.

IMPORTANT EVENTS

22 23


MAJOR FACILITIES

PERFORMANCE INDICATORS

•Industrial Technology Center

•National Seed Quality Testing Facility at CIMAP

•National Quality Referral Laboratory for MAPs and Herbal Products

•Installation of Inductively Coupled Plasma: A Facility for Precise Estimation of Trace and Heavy Metals

•Digital Photos and Art Lab

•Botanical Reference Substances (BRS)

CIMAP organized Thirty two entrepreneurship training programmes in different parts of the country through

which persons were provided technical knowledge and guidance for cultivation, processing, marketing etc. for

quality production of medicinal and aromatic plants.

One MAP journal (one volume; four numbers), six farm bulletins, two popular magazines (Hindi & English),

two information folders, two souvenirs, one-proceeding, one compilation of Institute Seminars (Gyanankur),

Annual Report Highlights, Annual Report,one Planner and two issues of Newsletter were published this year.

Two plant varieties were released and two new agrotechnologies were developed.

Thirteen sponsored projects completed during this financial year, thirty nine sponsored projects were

progressing according to plan and Six R&D projects and seven consultancy projects were taken up. CIMAP has

been working as nodal laboratory and participating laboratory in CSIR's network projects for mission mode

research.

A total of seventeen patents (twelve foreign) were granted to the Institute this year.

100 original research articles, 19 book chapters and 48 popular articles were published by S&T staff of CIMAP.

Forty three Seminar presentations and invited talks were made by the scientists of CIMAP.

Entrepreneurship Training Programmes

In House Publications

Plant varieties & Agrotechnologies

Sponsored Projects

Patents

Publications

In House Seminars

External Cash Flow

CIMAP earned an EBR of Rs 351.868 lakh, vis-à-vis CSIR grant of lab budget of Rs 1937.867 lakh and network

programme budget of Rs. 112.124 lakh.

•First Nina Saxena Excellence in Technology Award – 2007 to team CIMAP consisting of Dr.(s) SPS Khanuja,

Shilpi Paul, Ajit K. Shasany, Anil K. Gupta, Mahendra P. Darokar, M.M. Gupta, RK Verma, Govind Ram,

Anirudha Kumar, RK Lal, RP Bansal, Anil K. Singh, R.S. Bhakuni, Sudeep Tandon for development of

complete technology package for the variety of Artemisia annua by IIT, Kharakpur in their Foundation Day thProgramme on 18 August, 2007.

•Team CIMAP comprising of Vandana Srivastava, AS Negi, JK Kumar, MM Gupta, SPS Khanuja won the most

cited paper 2003-2006 award by Bioorganic & Medicinal Chemistry journal for their paper entitled, “Plant-

based anticancer molecules: A Chemical and biological profile of some important leads” published in

Bioorganic and Medicinal Chemistry, Volume 13, Issue 21(2005), pages 5892-5908.

•Team CIMAP comprising of Dr.(s) MM Gupta, GD Bagchi, Karuna Shankar, J Singh, SPS Khanuja received a

letter of appreciation from Dr. Nityanand, Chairman of Indian Pharmacopeia Commission, Govt. of India for

significant scientific contribution to IP-2007 by Team CIMAP on 11 August 2007 at CDHPC meeting of IPC at

Mumbai.

•CIMAP recognized as Designated National Repository (DNR) by National Biodiversity Authority (NBA).

•Dr. RS Sangwan elected Fellow of the National Academy of Sciences, Allahabad.

•Dr. DD Patra elected Fellow of the National Academy of Agricultural Sciences, New Delhi.

•Dr. DD Patra Awarded ISCA Platinum Jubilee Award, 2008 conferred by Indian Science Congress

Association, Kolkata

•Dr Dwijendra Singh elected fellow of the Royal Entomological Society, London.

AWARDS & RECOGNITIONS

24

Annual Report 2008 : Highlights

annual reports 2008

Documents