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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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ThesisItem 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 CGenerating 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.ThesisItem 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 ANutmeg (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 succeedThesisItem Open Access Management biodegradable plant tissue culture lab wastes through biomethanogenesisof(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikara, 2008) Abdulla Fayas, T; KAU; Rajendran, P CGenerating 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 crisis. 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, Methcmosarcina 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 formicicum with Methanosarcina barkeri. Physio-chemical characters like C/N ratio of the TC wastes, pH and temperature of medium and Hydraulic retention time was also observed for the various treatments. The C/N ratio of the TC wastes was found to be very low and nowhere near the optimum C/N 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 bio gas generation were found out to be the low C/N ratio of the TC waste and the low pH of the medium. The present study indicated the possibility of bio-gas generation from TC waste through fortification using various supplements like coconut water and coir pith which have higher C/N ratio.ThesisItem Open Access Morphomolecular charecterisation of the variants of piper nigrum L. variety panniyur -1(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2008) Smitha Bhasi; KAU; Swapana AlexThe study entitled “Morphomolecular characterization of variants of Piper nigrum L. variety Panniyur-1” was conducted at the Department of Plant Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram and in the Block V of Panniyur-1 at the Regional Agricultural Research Station (RARS), Ambalavayal during the year 2006-2007 with an objective of characterizing the variants of black pepper variety Panniyur-1 based on morphological traits and RAPD profiles. Black pepper often referred to as the ‘King of spices’ is the most important spice in the world. The first ever hybrid of black pepper, Panniyur-1 (Uthirankotta x Cheriyakaniyakadan) is the most popular pepper variety grown in India and also in Kerala. In black pepper, propagation through cuttings is being practiced for decades for producing true-to type plants. However, contrary to this belief, there are reports for the existence of variability. Variability was reported even at the intraclonal level. The first such report in black pepper was in the local variety Karimunda (Ratnambal et al., 1985). According to Pradeepkumar et al. (1999), there exists intra-clonal variability in yield among the hybrid clone Panniyur-1 at the RARS, Ambalavayal. Such reports deserve serious concern and in depth analysis as pepper is a leading commercial crop of India, important in the domestic as well as international markets. The present study was taken up in this context utilising the progeny of the forty variant plants reported by Pradeepkumar et al. (2003) from the RARS, Ambalavayal. The objective was to assess the extent of variability with respect to morphological traits including yield parameters as well as the molecular analysis of genetic variability. On morphological analysis of the forty plants, considerable variation was observed. The maximum variation was observed in number of berries per spike followed by drying percentage. The analysis of the dendrogram showed that none of the plants were 100 per cent similar at a distance of 1.0. At a distance of 2.0 the clones can be grouped into five clusters. At a distance of 10, the plants can be grouped into two clusters comprising a major group with twenty-nine plants and a minor group with eleven plants. Molecular analysis also revealed variability, accounting for 66.34 per cent polymorphism. In the dendrogram at the similarity index 0.70 the plants grouped into two major clusters indicating thirty per cent dissimilarity. None of the plants were showed 100 per cent similarity. All the forty plants under study formed individual clusters at a similarity index 0.91 except V36 and V37. Ninety percent similarity was observed between the plants V20 and V30. At a similarity index below 0.70 the dendrogram showed a cluster including all the plants except V14. The present findings need further confirmation with more number of primers and other molecular markers like ISSR, AFLP etc. The occurrence of variability among the clones of Panniyur-1 in other major pepper growing tracts also needs to be investigated in detail.ThesisItem 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 CGenerating 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.ThesisItem Open Access Agrobacterium tumefaciens mediated transfer of exogenous hydroxy methyl glutaryl CoA (HMG CoA) reductase gene to Centella asiatica L(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2008) Lekshmi, R S; KAU; Soni, K BA study on “Agrobacterium tumefaciens mediated transfer of exogenous hydroxyl methyl glutaryl CoA (HMG CoA) reductase gene to Centella asiatica L.” was conducted at the Department of Plant Biotechnology, College of Agriculture, Vellayani during 2006-2008. Centella asiatica is an important medicinal plant, which is used in many ayurvedic formulations. It contains a blend of compounds including triterpenes (asiatic acid, madecassic acid and asiaticoside), which are responsible for its medicinal properties. Among the various secondary metabolites in Centella asiatica, asiaticoside possesses remarkable pharmaceutical value due to its anti-inflammatory, antitumour, neuroprotective, skin care and toning effects. Since the asiaticoside content in the Indian ecotypes is less, the industrial demands are met by importing this plant from African countries. By improving asiaticoside content, utilization of Indian ecotypes could be improved and thereby the cost of medical preparations could be reduced. Metabolic engineering is becoming a popular approach for the modification of medicinal plants for altering the metabolite content. Medicinal plants with quantitatively and qualitatively improved pharmacological properties have been produced by metabolic pathway engineering. The present study was undertaken with an objective to enhance the asiaticoside content by introducing exogenous hmgr gene to Centella asiatica. This gene is responsible for coding hydroxy methyl glutaryl CoA reductase enzyme which acts at the upstream of the triterpene biosynthetic pathway and produce mevalonic acid. Callus was induced from leaf and node explants of Centella and MS medium supplemented with Kn 4 mg l-1 and NAA 2 mg l-1 was found to be the best for callus induction. Callus initiation was faster (23 days) from node compared to leaf (25 days), with 100 and 92.85 per cent induction respectively. Of the various media tried for regeneration, the highest regeneration (0.052%) was obtained on MS medium supplemented with Kn 4mg l-1 and NAA 2 mg l-1. Agrobacterium tumefaciens strain EHA105 harbouring the plasmid pBE2113 containing nptII and hmgr gene was used for transformation. The sensitivity of Agrobacterium strains and Centella callus to different concentrations of kanamycin was evaluated. The lethal doses of kanamycin to Agrobacterium and Centella callus were 350 and 125 mg l-1, respectively. The effective dose of cefotaxime for the elimination of bacteria was 50 mg l-1 and the lethal dose of cefotaxime to callus was 100 mg l-1. Genetic transformation was achieved by co-cultivating callus with bacterial suspension (OD600 of 0.1) containing 100 μM acetosyringone. An infection time of 20 min and co-cultivation period of four days were given. The transformed tissues were selected on MS medium containing 200, 250 and 300 mg l-1 of kanamycin. The survival of the tissues on these media after three weeks was 18.75, 19.35 and 13.63 per cent, respectively. Transformation was confirmed by PCR analysis with npt II gene specific primer. All the three samples gave appreciable quantity of the product of size 700 bp which was comparable to positive control. In the present study regeneration of the transformed tissues was not obtained in the media standardized. Even though an attempt was made to analyse asiaticoside content in the transformed callus using thin layer chromatography (TLC), there was no detectable quantity of asiaticoside. This could be due to the undifferentiated nature of the callus. As the accumulation of asiaticoside is mainly in the leaves, a better protocol for regeneration needs to be developed so that further studies on triterpenoid analysis could be carried out.ThesisItem Open Access Polymerase chain reaction based detection of banana bunchy top virus(BBTV) and banana streak virus(BSV)(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2008) Ponsubrya, R K; KAU; Swapna, AlexBanana is an important fruit crop of Kerala. It is affected by various diseases including viral diseases, of which Banana Bunchy Top Virus (BBTV) and Banana Streak Virus (BSV) assume economic importance. Detection of these viruses in early stage warrants great significance. The present study entitled “Polymerase Chain Reaction based detection of Banana Bunchy top virus (BBTV) and Banana streak virus (BSV)’’ was taken up in this context. It was carried out at the Department of Plant Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram during October 2006 to July 2008 with the objective of developing a protocol for the detection of these two DNA viruses of banana using polymerase chain reaction (PCR) technique. DNA from infected samples of BBTV and BSV as well as healthy plants was isolated using CTAB and modified CTAB methods. In terms of quality and quantity of DNA isolated, both the methods were good. The isolation of the genomic DNA using the CTAB and modified CTAB method was also standardized. Polymerase Chain Reaction was carried out using designed as well as reported primers. DNA sequence used for the primer designing was taken from National Centre for Biotechnology Information. The primers were designed for the coat protein gene of the two viruses viz., BBTV and BSV. The primer designing was done manually using certain fixed parameters such as primer length, GC content, melting temperature etc. The length of the designed primers ranged from 16nt to 24nt and Tm from 51.7oC to 62.9oC. Designed primers viz., Af and Ar in the case of BBTV and Bf and Br in the case of BSV were analysed using the oligocalc-oligonucleotide calculation program and the similarity search was done using the BLASTN. The result of the BLASTN showed that the primer sequences were similar not only to banana but also to other organism like Arabidopsis thaliana, Drosophila melanogaster etc. The reported primers were obtained from Su et al. (2003) in the case of BBTV and Braithwaite et al. (1995) in the case of BSV. The amplification gave a band of about 500bp in the case of BBTV with the primer Af and Ar and 1000bp in the case of BSV with the primers Bf and Br and 1Kb in the case of the reported primers of BBTV. There was no amplification in the BSV samples using reported primer as well as using IC-PCR technique. The present study made it possible to detect the presence or absence of the viruses through PCR. More number of samples and primers can be used in the future to detect the virus. This technique can also be extended to other viruses also.ThesisItem Open Access Molecular diversity of pseudomonas fluorescens antagonistic to ralstonia solanacearum(College of Horticulture, Vellanikkara, 2008) Sindhu, C Cheeran; KAU; Girija, DThe study on “Molecular diversity of Pseudomonas fluorescens antagonistic to Ralstonia solanacearum” was conducted at the Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, during the period from August 2006 to May 2008. Pseudomonas fluorescens is an efficient bacterial antagonist effective against various soil-borne plant pathogens. The present investigation was carried out to assess the diversity of P. fluorescens isolates from the Western Ghats of Kerala and their antagonistic activity against Ralstonia solanacearum. Fourteen isolates of fluorescent psuedomonads tentatively identified as Pseudomonas fluorescens were isolated from different areas coming under the Western Ghats of Kerala. The isolates were maintained in sterile water, as stabs and also as glycerol stocks. Diversity of these isolates were studied by various morphological, physiological and biochemical tests. The antagonistic activity of these isolates was tested against Ralstonia solanacearum, which causes bacterial wilt in tomato. Five potential isolates which showed good antagonism under in vitro conditions, were selected and their efficacy was tested using tomato plants under pot culture condition. One of the native isolates Pf-3 collected from Malappuram tract was found to be effective in controlling wilt incidence. Molecular diversity was analyzed using Random Amplified Polymorphic DNA (RAPD) and rep-PCR. RAPD analysis was carried out using 10 different primers. All the primers except OPN 4 showed 100 per cent polymorphism. For OPN 4, a unique monomorphic band was amplified in all the isolates. The genetic similarity matrix gave the highest value of 0.92 between pf-1 and Pf-2 isolate followed by 0.90 between Pf-1 and Pf-3. In rep-PCR genomic fingerprinting, DNA primers complementary to highly conserved repetitive DNA sequences, present in the genomes of bacteria are used for amplification. Four different rep primers were used in the present analysis. The similarity coefficient was 1.0 for Pf-1 and Pf-2 followed by 0.96 for Pf-3. High degree of diversity was observed among the isolates at molecular level as detected by RAPD and rep-PCR. Even though, the three isolates Pf-1, Pf-2 and Pf-3 (collected from Malappuram) were grouped in a single cluster in all the dendrograms, slight differences were seen in the gel profiles indicating that they were not genetically identical. The results indicate that diversity among bacterial isolates within a single species, not detected by routine tests like cultural, morphological, physiological and biochemical characterization can be assessed by using molecular tools like RAPD and rep-PCR.ThesisItem Open Access Characterization of proteins and trehalose as heat shock regulators in insect system(College of Horticulture, Vellanikkara, 2008) Swetaleena, Tripathy; KAU; Mani, ChellappanThe study entitled “Characterization of proteins and trehalose as heat shock regulators in insect system” was undertaken in the laboratory of Centre for Plant Biotechnology and Molecular Biology and Department of Entomology, College of Horticulture, Kerala Agricultural University in Vellanikkara to study the underlying mechanism of the red flour beetle, Tribolium castaneum Herbst. (Tenebrioniidae: Coleoptera) to resist stress. Tribolium castaneum had been seen as a serious pest of flour mills and food processing industries causing huge loss every year. The use of elevated temperatures had long been recognized as an effective strategy for managing the dreaded stored product insect. The major objectives of the study were to find out the thermal death stage of the red flour beetle when exposed to a particular temperature and time period at which the insect was unable to combat stress further and to characterize the metabolites responsible for the survival of this insect during anhydrobiosis. The mass culturing of the insect was done at a temperature of 28°C with 70 per cent relative humidity. The population built up was faster on the wheat flour (30.97±0.20) followed by the combination of both wheat flour and semolina (30.91±0.15) while it was less in semolina alone (30.29±0.13). At the same time, the per cent contamination and per cent mortality were significantly more on wheat flour (41.3±3.53 and 19.58±0.93) when the relative humidity was higher (88%). The per cent contamination and per cent mortality were positively correlated with the temperature and relative humidity. The insect completed its life cycle within 34-38 days in the present study. Different stages of insects (neonates, V instar grub, pupae and the adult beetles) were subjected to temperatures ranging from 35°C to 60°C at an incremental increase of 5°C for a period of 1 h, 2 h and 4 h respectively. The insects were not killed even at 4h exposure to the above temperatures. However, the mortality of each stage of the insects was increased with the exposure time and the incremental temperature increase. A higher mortality per cent was observed at 50°C for 8 h (98.7±0.33) and 55°C for 6 h (90.7±0.67) respectively. Complete mortality of the insect was observed at 55°C and 60°C when the insect stages were exposed to the temperature for 7 h and 5 h respectively. Subjecting the insects to stress showed accumulation of a heat shock protein (hsp) along with an insect sugar called trehalose. It was observed that the heat shock protein and trehalose content in insects increased significantly with the stress. When the insects were exposed to 50°C, the corresponding protein content was in the order of neonate (36.81 mg/g) < pupa (38.16 mg/g) < adult (39.23 mg/g) < V instar grub (40.81 mg/g). Thus, the decrease in the total protein content was observed to be more for the neonate at 50°C when compared to the other stages. Similarly, when the insects were subjected to heat stress at 35°C, the trehalose content in the neonate sample was (14.97 mg/g) and it was significantly higher in V instar grub (16.49 mg/g), pupa (17.64 mg/g) and the adult (18.48 mg/g). But there was significant decrease in the trehalose content of the insect after the recovery of the stress period. The molecular weight (70KDa) of the ‘Trib hsp’ was determined from the SDS-PAGE analysis and its identity was further confirmed by the N-terminal sequencing. It showed 80 per cent homology with the heat shock protein. Further theoretical analysis of the protein sequence showed that the protein was stable and it was composed of four conserved domains. The data generated so far could be useful in developing a heat control strategy to manage this insect. Further studies with respect to characterization of anti heat shock protein could also be carried out in order to see its effect on a heat shock cognitive. The characterization of the stress regulators would help to manage the public health pest’s viz., mosquito. The genes coding for stress regulatory proteins could also be isolated, cloned and used for development of genetically modified insects.