Kerala Agricultural University, Thrissur

The history of agricultural education in Kerala can be traced back to the year 1896 when a scheme was evolved in the erstwhile Travancore State to train a few young men in scientific agriculture at the Demonstration Farm, Karamana, Thiruvananthapuram, presently, the Cropping Systems Research Centre under Kerala Agricultural University. Agriculture was introduced as an optional subject in the middle school classes in the State in 1922 when an Agricultural Middle School was started at Aluva, Ernakulam District. The popularity and usefulness of this school led to the starting of similar institutions at Kottarakkara and Konni in 1928 and 1931 respectively. Agriculture was later introduced as an optional subject for Intermediate Course in 1953. In 1955, the erstwhile Government of Travancore-Cochin started the Agricultural College and Research Institute at Vellayani, Thiruvananthapuram and the College of Veterinary and Animal Sciences at Mannuthy, Thrissur for imparting higher education in agricultural and veterinary sciences, respectively. These institutions were brought under the direct administrative control of the Department of Agriculture and the Department of Animal Husbandry, respectively. With the formation of Kerala State in 1956, these two colleges were affiliated to the University of Kerala. The post-graduate programmes leading to M.Sc. (Ag), M.V.Sc. and Ph.D. degrees were started in 1961, 1962 and 1965 respectively. On the recommendation of the Second National Education Commission (1964-66) headed by Dr. D.S. Kothari, the then Chairman of the University Grants Commission, one Agricultural University in each State was established. The State Agricultural Universities (SAUs) were established in India as an integral part of the National Agricultural Research System to give the much needed impetus to Agriculture Education and Research in the Country. As a result the Kerala Agricultural University (KAU) was established on 24th February 1971 by virtue of the Act 33 of 1971 and started functioning on 1st February 1972. The Kerala Agricultural University is the 15th in the series of the SAUs. In accordance with the provisions of KAU Act of 1971, the Agricultural College and Research Institute at Vellayani, and the College of Veterinary and Animal Sciences, Mannuthy, were brought under the Kerala Agricultural University. In addition, twenty one agricultural and animal husbandry research stations were also transferred to the KAU for taking up research and extension programmes on various crops, animals, birds, etc. During 2011, Kerala Agricultural University was trifurcated into Kerala Veterinary and Animal Sciences University (KVASU), Kerala University of Fisheries and Ocean Studies (KUFOS) and Kerala Agricultural University (KAU). Now the University has seven colleges (four Agriculture, one Agricultural Engineering, one Forestry, one Co-operation Banking & Management), six RARSs, seven KVKs, 15 Research Stations and 16 Research and Extension Units under the faculties of Agriculture, Agricultural Engineering and Forestry. In addition, one Academy on Climate Change Adaptation and one Institute of Agricultural Technology offering M.Sc. (Integrated) Climate Change Adaptation and Diploma in Agricultural Sciences respectively are also functioning in Kerala Agricultural University.

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Open Access
Forecasting volatility of the indian stock market
(Department of Rural Banking and Finance Management, College of Co-operation, Banking and Management, Vellanikkara, 2009) Khadilkar Guruprasad, Hari; KAU; Molly, Joseph
The present study on ‘forecasting volatility of the Indian stock market’ was conducted with the main objectives of examining the volatility behaviour of the Indian stock market, to forecast the sector- wise volatility of the Indian stock market and to identify the most efficient volatility forecasting model among the different models used. For the study the biggest stock market in India in terms of total turnover and volume of transactions, ie, National Stock Exchange was selected. For analyzing the volatility behaviour of the Indian stock market as a whole, S&P CNX Nifty index was taken. Five companies representing five different sectors were selected for forecasting sector – wise volatility. The study used secondary data on daily close prices of individual stocks from November 1994 to October 2008, and for Nifty, daily close values, from November 1995 to October 2008 from the website of National Stock Exchange, www.nseindia.com. The study revealed presence of strong volatility in the Indian stock market. The histogram drawn for the volatility of all samples showed that the distribution of volatility was not normal. There was positive skewness and all the distribution of volatility was leptokurtic. This proved the presence of high peak values (squared returns) in the sample data, exposing the evidence of volatility clustering and the possibility for prediction of future volatility. While analysing sector -wise volatility, the diversified sector represented by Reliance Industries Limited showed the highest volatility compared to that of Nifty and the other sectors. In other words, Reliance is the most volatile stock among the samples selected for the study. Reliance and Infosys had good predictability of volatility in the stock market. The best identified model for forecasting the volatility of stock markets is the EWMA. Then comes AR (1) followed by MA (3), RWM and HMM. Random walk model was found suitable for the prediction of volatility of two sectors - IT (Infosys) and engineering heavy (BHEL) only. But the MAPE values of these were high. Historic mean model could not predict the volatility in the stock market with precision, for the index as well as for any of the five companies. Out of three, six, nine and twelve monthly moving averages taken for predicting the volatility three months moving average was found most suitable for all the samples. Prediction of volatility using the most efficient model of EWMA identified indicated decreasing trend of volatility for the next six months, except for Infosys. The confidence limits for the Nifty and the stocks of five companies based on volatility for the sample period found that for Infosys the distribution of volatilities for the out of sample period are coming within the prefixed UCL and LCL and it ensures that the volatility is under control and predictable with high degree of precision. The ever increasing market segments, advancement of technology, widening market reach and multi dimensions of stock market provide ample scope for further research in this area to the advantage of the investors and other market participants.
Open Access
Effect of seed treatments and shade on seedling growth dynamics of pongamia pinnata (Linn.) pierre in the nursery
(Department of Forest Management and Utilisation, College of Forestry, Vellanikkara, 2009) Gururaj Sankeshwar, B; KAU; Gopakumar, S
The present study entitled “Effect of seed treatments and shade on seedling growth dynamics of Pongamia pinnata (Linn.) Pierre in the nursery” was carried out in College of Forestry, Kerala Agricultural University, Vellanikkara, Thrissur during the period of 2007-2009. In the first phase, seeds were subjected to six pre-sowing treatments. GA3 treated seeds recorded highest Germination percentage, Final Mean Daily Germination (FMDG), Germination Value (GV) and Peak Value (PV). The effect of seed treatments were observed on various biometric parameters like height, collar diameter, number of leaves and primary branch. GA3 treated seeds recorded better height and collar diameter during the initial stage of seedling growth. At the end of study cold water seed treatment was found to be effective in enhancing seedling growth. However, none of the pretreatments was observed to cast a significant effect on biomass production and root growth parameters in Pongamia pinnata seedlings. In the second phase, to evaluate seedling growth under different shade conditions, seedlings were placed under 25 per cent shade, 50 per cent shade, 75 per cent shade and full sunlight situations. Shade treatments did not significantly influence height and collar diameter growth of Pongamia pinnata seedlings in the nursery. The maximum leaf production and primary branch production was observed in seedlings kept under full sunlight and 50 per cent shade respectively. Under seventy five per cent shade seedlings produce maximum shoot and root dry weight. The shoot: root ratio and total biomass was also maximum for seedlings kept under 75 per cent shade. Root length and spread were maximum under 50 per cent shade. However, shade did not significantly effect secondary root production. Shade levels also did not influence Pongamia pinnata seedlings with respect to relative growth rate and leaf area. However, Specific leaf area and Leaf area ratio was higher under 25 per cent shade. Highest Specific leaf weight was observed for seedlings under full sunlight. Shade had no significant effect on leaf weight ratio of seedlings. Maximum stomatal number was recorded under 25 per cent shade. In the nursery stage shade is casting an overriding influence with regards to various growth parameters of Pongamia pinnata.
Open Access
Performance analysis of clove bean ipomoea muricata (L) jacq. genotypes
(Department of Olericulture, College of Horticulture, Vellanikkara, 2008) Malsawmkimi; KAU; Salikutty, Joseph
An experiment was carried out in the Department of Olericulture, College of Horticulture, Kerala Agricultural University, Vellanikkara during August 2007 - January 2008 to analyze the performance of different accessions of clove bean Ipomoea muricata (L.) Jacq. The major objectives of the study were to genetically catalogue the germplasm and to assess the genetic variability, divergence, heritability, genetic gain and correlation of different traits with yield. Twenty five accessions collected from different parts of Kerala were grown in randomized block design with three replications. Cataloguing of the germplasm evidenced significant differences for the characters like vine length, days to first flower production, days to first harvest, length and weight of pedicel, length of fruit, yield per plant, duration of the crop, number of harvests and contents of vitamin C, calcium and crude fibre. There was also much variability in the germplasm with regard to colour shades of vine, flower and fruit. The accession IM-14 was found to be the highest yielder (4.24kg/plant) coupled with maximum length (3.675cm), girth (4.49cm) and weight of pedicel (93.27g) and length (2.8cm), girth (6.61cm) and weight of fruits (3.27g). Maximum vine length was observed in IM-7 (414.48cm). The accession IM-15 was the second best yielder (4.13 kg/plant) with superior yield contributing characters. Among quality attributes, protein content was found to be highest in IM-21, phosphorus in IM-2 and crude fibre in IM-10 and IM-3. The iron content ranged from 0.13mg/100g to 0.19mg/100g, vitamin C ranged from 24mg/100g to 48 mg/100g and calcium content ranged from 230.67mg/100g to 209.79mg/100g. Highest genotypic and phenotypic coefficients of variation were observed for weight of pedicel, yield per plant and crude fibre content. High heritability coupled with genetic gain was observed for yield per plant. The 25 accessions were grouped into six clusters and no parallelism between geographical distribution and genetic diversity was observed. Strong association was revealed between yield and length, girth and weight of pedicel and weight of fruit. Highest positive direct effect on yield was contributed by weight of individual fruits. A selection model was also formulated using characters like length, girth and weight of pedicel, length and girth of fruit, days to first flower production, vine length, protein, phosphorus, iron, ascorbic acid and crude fibre. Based on selection index accession IM-14 was identified as the best performer followed by the accessions IM-15, IM-12 and IM-11
Open Access
Population dynamics and management of erythrina gall wasp quardrastichus erythrinae kim.
(Department of Agricultural Entomology, College of Agriculture, Vellayani, 2008) Beena, M P; KAU; Thomas Biju, Mathew
. Studies were conducted to understand the population dynamics and management of Erythrina Gall Wasp (EGW)Quadrastichus erythrinae. An abstract of the work done and the results are given below. Population dynamics of Q.erythrinae was studied by observing weekly emergence of male and female insects from leaflets and petioles collected from field during the period from January 2006 to June 2007. The results showed that population was high in the months of July 2006, January 2007, April 2007 and June 2007.The present study indicated that the emergence od adult EGW was the highest from petioles. The increase in weight due to EGW infestation and gall production was the highest in the left lateral leaflets. Weekly mean population of EGW emerged during the period of study showed that the total number of male EGW (1096.36) was higher than the female insects emerged (763.13). Weekly mean population of the male and female emerged from the leaflets and petioles was correlated with weather parameters viz. maximum and minimum temperature. Maximum and minimum relative humidity and rainfall prevailed during the time of sampling, previous one week, previous 14 days,week 21 days before and week 28 days before. The results showed significant and positive correlation between population of male EGW and the minimum temperature, while no correlation existed between female population and other parameters studied. Comparison of biometric observation of uninfested and infested plant parts indicated that the weight of the leaflets and petioles increased significantly due to an increase in biomass. Due galling by EGW, the girth of the petioles was shortened to about one third of uninfested petioles. The emergence of EGW was maximum from petioled scored in the damage category ‘9’. Among the four groups of Erythrina stricta Roxb..stricta trees studied, based on the height of the trees, no group turned out to withstand the damage by EGW. Thiomethoxam, imidacloprid, carbaryl and acephate proved superior among the chemical tested insecticides tried against female and male EGW in the laboratory . Neem oil 2 per cent and neem oil-garlic 2 per cent proved superior among the botanical insecticides tried for protecting the newly formed side shoots. Among the systemic /translaminar insecticides applied on the galled plant parts, triazophos 0.05 per cent and imidacloprid .002 per cent were significantly superior in controlling the EGW emergence. In the field experiment conducted to evaluate insecticide-fungicide combination for EGW infestation and protection of the new side shoots, carbaryl 0.1 per cent and combination of imidacloprid 0.002 per cent and carbendazim 0.2 per cent effectively protected the side shoots up to 8 weeks after spraying. Residue analysis of dry pepper berries collected from pepper vines trailed, on the insecticide sprayed crythrina standards showed that residues at the time of harvest from trees sprayed with acephate 0.05 per cent dimethoate 0.05 per cent were below detectable limit. Where as the residues in pepper collected from lamda cyhalothrin 0.05 per cent and triazophos 0.05 percent treated trees was 1.38 ppm and 1.16ppm respectively. An overall analysis and interpretation of the data on population in relation to phenology to E.stricta foliar growth showed that there existed a six monthly cycle of damage by gall wasp resulting in complete defoliation of the trees by about six month followed by the next cycle of regrowth and re infestation .
Open Access
Reproductive biology of macrobrachium canarae ( tiwari, 1958 ) (decapoda, palaemonidae)
(Department of Fishery Biology, College of Fisheries, Panangad, 2008) Sreedevi, K H; KAU; Jayachandran, K V
The aim of this project is to introduce the freshwater prawn Macrobrachium canarae (Tiwari, 1958) as a candidate species in a community aquarium. The orange red shining spot on the second cheliped where the movable finger joins with palm makes it appealing in the aesthetic sense. This species is found to breed in captivity also. For the effective management of the prawn in the aquarium, a thorough knowledge on the various aspects of its reproductive biology is a prerequisite. The various aspects of reproductive biology dealt with are 1. Taxonomy 2. Sexual dimorphism 3. Breeding dress 4. Maturity stages and ovarian development 5. Fecundity 6. Moulting Since the species do not exhibit sexual dimorphism in size, it is difficult to identify male and female. The difference in colour of antennular flagellum between sexes becomes useful especially when they are bred in captivity. The data on maturity stages and also the time of ovarian development finds its use especially in the aquarium rearing. The fecundity studies gives an idea of the number of offsprings which could be produced from a single brood .
Open Access
Ovararian maturation, breeding and early embryonic development of an indigenous ornamental cyprinid of the western ghats - chela fasciata silas
(Department of Fishery Biology, College of Fisheries, Panangad, 2008) Indira, Divipala; KAU; Anna Mercy, T V
Chela fasciata is an indigenous ornamental cyprinid endemic to the Western Ghats of Kerala. It inhabits the riffle zones of River Bharathapuzha at Thootha. A total of 144 fishes (94 females and 50 males) ranging in size from 26.0 to 82.0 mm were collected from the wild and used for the study of reproductive biology. The gonads were quantified into six maturity stages based on external morphology as immature virgin, maturing virgin, early ripening, late ripening, ripe and partially spent. Studies on oocyte distribution of the six stages of maturity were done to understand the maturation of ova in the ovary. The ovaries of Chela fasciata showed asynchronous oocyte development. The oocyte development was classified into nine different oogenic stages namely, chromatin nucleolus stage, early perinucleolus stage, late perinucleolus stage, yolk vesicle stage, primary yolk stage, secondary yolk stage, tertiary yolk stage, migratory nucleus stage and mature oocyte). The first mature females appeared in the length group of 40 – 45 mm and males in the group of 25 – 30 mm. The size at first maturity for female was found to be at 45.75 mm TL (45 – 50 mm) and for males at 36.25 mm TL (35 – 40 mm). All the females were mature by 60.00 mm TL and the males by 45.00 mm TL. Based on the ova diameter frequency study of the ripe ovary, Chela fasciata was found to be a multiple spawner, with a protracted spawning season, the individuals spawning intermittently. Absolute fecundity of the fishes ranged from 2669 to 4437 in fishes of size range 49.5 mm to 82 mm TL. The number and size of eggs were found to be directly proportional to the size and age of the fish. Fecundity showed a positive linear relationship (5% level of significance) with the length and weight of both the fish and ovary. The embryonic development studies showed that the cleavage stage of the egg lasted for about 2 hrs 10 min., post fertilization. The inside egg embryo stage had lasted for 21 hrs, post fertilization. The eggs hatched at the end of 21 hrs, and the embryos emerged tail first. Pigmented eye was seen in one-day-old free embryo. Free embryo stage lasted for three days post fertilization. By the end of third day, the mouth developed, the yolk sac was empty and the exogenous feeding started, the free embryo entering the larval period. A complete understanding of basic reproductive biology of this fish will definitely help in the commercial production under captivity for the domestic and export market of this indigenous ornamental fish.
Open Access
Management of biodegradable plant tissue culture lab wastes through biomethanogenesis
(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2008) Abdulla Fayas, T; KAU; Rajendran, P C
Generating renewable source of energy from tissue culture laboratory waste by the process of biomethanogenesis is the focal theme of present investigation. Unlike developed countries, the developing countries are hesitant to establish more number of biotechnology/ tissue culture laboratories due to financial constraints. Easy and regular availability of biogas from TC wastes will be a boon to establish self-sustainable TC laboratory in view of present energy The biogas experimental units required for the study was designed and various treatments were employed for the biodegradation of tissue culture waste, using the methanogenic bacteria Methanobacterium ruminatium, Methanobacterium formicicum; Methanosarcina barkeri, Bactereoides ruminicola, Selenomonas ruminatium, Eubacterium tortuosum and Clostridium butyricum. Treatment involving TC waste and cow dung was also conducted for biomethanation in the present study. Quantity of gas production and its combustibility was noticed for various treatments. In bacterial treatments the quantity of gas generation was highest for Clostridium butyricum. Only treatments involving cow dung produced combustible gas. Molecular characterization, of methanogenic bacterial cultures was also done for finding the genetic similarity between them. RAPD followed by scoring . of the bands by UPGA routine showed maximum similarity between bacterial cultures of Methanobacterium ruminatium and Methanobacterium jormicicum with Methanosarcina barkeri. Physio-chemical characters like CIN ratio of the TC wastes, pH and temperature of medium and Hydraulic retention time was also observed for the various treatments. The CIN ratio of the TC wastes was found to be very low and nowhere near the optimum CIN ratio of 20-30 required for gas production. Other parameters like pH of the treatments and Hydraulic retention time was also. • noticed. The pH of the treatments involving bacterial cultures was very low, considering the normal pH of 6.8 to 7.5 required in biogas generation. The main constraints in the biogas generation were found out to be the low CIN ratio of the TC waste and the low pH of the medium. The present study indicated the possibility ofbio-gas generation from TC waste through fortification using various supplements like coconut water and coir pith which have higher CIN ratio.
Open Access
In vitro shoot regeneration and micrografting in nutmeg (Myristice fragrans houtt.)
(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2008) Liffey Zachariah, Antony; KAU; Valsala, P A
Nutmeg (Myristica fragrans Houtt.), is dioecious and dimorphic in branching habit with erect growing orthotropic and horizontally growing plagiotropic shoots. The long gestation period and dioecious nature of the crop causes difficulty in the production of quality planting materials of known sex. Vegetative propagation, budding and grafting with orthotropic scion material produces erect growing tree with upright tree architecture. In vegetative propagation, scarcity of orthotropic scion material is a limiting factor in large scale production of planting materials. So the programme “In vitro shoot regeneration and micrografting in nutmeg (Myristica fragrans Houtt.)” was taken up. The objectives of the study were: (1) To identify the culture conditions for multiple shoot production from orthotrops of gynoecious plants of nutmeg through enhanced release of axillary buds and shoot tip culture and (2) To standardize micrografting technique with in vitro and in vivo shoots as scion. The work was done at CPBMB, College of Horticulture, Vellanikkara. SH medium (Schenk and Hildebrandt, 1972) was found to be the best basal medium for in vitro culture establishment of nodal segments of nutmeg compared to ½ MS (Murashige and Skoog, 1962) and WPM (Lloyd and Mc Cown, 1980). Surface sterilization of nodal segments by soaking in (0.1%) carbendazim for 10 minutes followed by (0.1%) HgCl2 treatment for six minutes and sterile water wash, recorded 33% survival of cultures. The best explant for culture initiation was nodal segments. The best season for culture establishment was summer months (April-May) compared to rainy season (June- July). Loss of cultures was due to fungal contamination and necrosis of tissues. The media combination SH + Thidiazuron (TDZ) 0.03mg l-1 + Activated Charcoal (A.C.) 0.5% recorded bud expansion in 50% of the cultures within a period of nine days. Nodal segments are superior to shoot tips in culture establishment. Culture condition for culture establishment was 26 ± 20C at a light intensity of 1000 lux. The carbon source; 2% Sucrose + 1% glucose or 5% sucrose supported bud elongation and leaf expansion Refinement of culture establishment media was attempted with organic supplements; Coconut water (5, 10, 15 and 20% v/v), Casein hydrolysate (10, 25 and 50mg l-1) and Brassinolide (0.05, 0.1 and 0.2mg l-1) with nodal segments from juvenile seedlings, regenerants from coppiced trees and mature trees. In explants from juvenile seedlings and mature trees, 5 to 15%coconut water supported culture establishment. In juvenile explants, shoot elongation was also observed at 10% coconut water. Casein hydrolysate supported bud expansion in juvenile and mature tree explants at 10 to 50 mg l-1. Bud elongation and leaf expansion was observed at 25 mg l-1 concentration. Brassinolide (0.2 mg l-1) supported bud expansion in juvenile explants. The suggested media for explants from juvenile as well as coppiced trees for culture establishment of nodal segments of nutmeg is SH + 0.03 mg l-1 TDZ + 25 mg l-1 Casein hydrolysate + 2% sucrose + 1% glucose + 0.5% A.C. Casein hydrolysate concentration for explants from mature trees could be 50 mg l-1. In vitro seed germination was observed in mature seeds in presterilized bottles with water soaked cotton/ little water. Somatic embryos were formed at the cut portion of six month old seeds in the medium of ½ MS + 2% Sucrose + A.C. 0.5%. Proliferation of callus and somatic embryos was observed with the medium B5 + 0.1 mg l-1 Kin + 0.01 mg l-1 NAA + 0.01 mg l-1 GA3 + 10.0 mg l-1 Casein hydrolysate + . 0.5% A.C. The response was obtained two and a half months after inoculation. Three days old in vivo germinated seedlings did not established under in vitro condition even though surface sterilization treatment with 0.1% Emissan for thirty minutes followed by 0.1% HgCl2 for six minutes was given. Feasibility of grafting in juvenile plants was studied with epicotyl grafting and got 80% success. Grafting was done on twenty days old seedling with scion material from different seedling. In vitro epicotyl micrografting was done with in vitro raised scion and root stocks. It was also done on in vivo germinated seedlings after surface sterilization. Scion shoot of 2.5 cm length was grafted on twenty days old root stock. Graft was cultured in liquid nutrient medium and survived for two weeks. Later fungal contamination destroyed the cultures. Grafting with in vitro shoots on in vivo raised root stocks did not succeed
Open Access
Exploitation of invitro cultures of Indian Madder(Rubia cordifolia.Linn) for anticancerous compounds
(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2009) Labade, Dinesh Sitaram; KAU; Asha Sankar, M
The present investigation on “Exploitation of in vitro cultures of Indian Madder (Rubia cordifolia L.) for anticancerous compounds” was carried out at the Plant Tissue Culture Laboratory of the Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara and Amala Cancer Research Centre, Thrissur during the period 2006-2008. The study was undertaken with the objective to standardize the in vitro techniques for initiation and proliferation of static and suspension cultures of Rubia cordifolia and to screen the in vitro cultures for synthesis of naphthoquinone and quantify it. It was also envisaged to enhance the level of product synthesis in in vitro cultures and to assess the anticancerous activity of in vitro and in vivo extracts in terms of cytotoxicity, antioxidant and prooxidant activities in vitro. Leaf, nodal and root derived callus cultures of Rubia cordifolia were established in vitro. Explants were pre treated with the fungicide, Bavistin 2.5 per cent for 15 minutes. Surface sterilization with mercuric chloride (HgCl2) at 0.1 per cent for 1 min and 30 sec was effective for yielding healthy, contamination free cultures from nodal segments and leaves, respectively. MS medium at full strength, supplemented with NAA at 2 mg l-1 along with BA at 0.5 mg l-1 was observed ideal for initiation and proliferation of calli. The auxin synergist phloroglucinol, when supplemented to the medium, did not not yield encouraging results, with respect to callusing in the experimental species. Root derived cultures were inferior with respect to callus initiation and proliferation, registering low values for all the parameters studied. Incubating in vitro cultures under illuminated condition at 26 ± 2 C was superior to dark incubation, with respect to callus initiation and proliferation. Chloroform – methanol at 8.5 :1.5 ratio was indentified as the appropriate solvent system for detection of naphthoquinone on thin layer chromatograms in the test extracts of the experimental species, with alcoholic KOH (10 per cent) as the spray reagent. Ms medium at full strength, fortified with NAA and BA at 2.0 mg l-1 and 0.5 mg l-1 respectively, which recorded maximum naphthoquinone synthesis, was standardized as the production medium. Enhancing concentration of sucrose to 5 per cent in the production medium, did not elicit a positive response on naphthoquinone production in vitro. Reducing nitrate concentration of the production medium, to half and one fourth the original concentration, resulted in enhanced in vitro synthesis of the target compound. Supplementing the production medium with yeast extract (1 per cent and 2 per cent) as well as precursor feeding with phenyl alanine and tyrosine each at levels of 50 mg l-1, 100 mg l-1 and 150 mg l-1 exerted a favourable influence on synthesis of naphthoquonines, in vitro. Incubation in dark resulted in marginal increase in in vitro production of naphthoquinones. Incorporation of autoclaved mycelia of Pythium aphanidermeatum at levels of 2.0 per cent and 5.0 per cent resulted in enhanced in vitro production of naphthoquinone. The abiotic elicitor, salicylic acid at concentration of 10 μM and 100 μM resulted in maximum synthesis of naphthoquinones in in vitro root cultures (8.76 units g -1 calli) of Rubia cordifolia. Immobilization of test calli with sodium alginate – calcium chloride complex as well as subjecting the in vitro cultures to stress conditions, as imposed by sorbitol failed to bring about an enhancement in the in vitro production of naphthoquinones. None of the explants employed in the study induced hairy roots, when co- cultured with the Agrobacterium rhizogenes strains, MTCC 2364 and MTCC 532. Based on cell count, subculturing intervals of leafs, nodal and root derived suspension were fixed as 24, 27 and 27 days respectively with the respective packed cell volume as 0.93 per cent, 0.83 per cent and 0.80 per cent. Naphthoquinone was detected, in ex vitro and in vitro test extracts at all levels of maturity tested. Both ex vitro and in vitro root extracts exihibited maximum cytotoxicity, as revealed by the percentage of cell death on DLA and EAC cell lines as well as their IC50 values. As compared to whole plant extract, in vitro systems of the experimental species exhibited least antioxidant action. Extent of pro-oxidant activity was higher in in vitro root extract of the experimental species.

Kerala Agricultural University, Thrissur

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