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Since ancient times, agriculture is an outdoor or open field production of crops. Open field production is climate and weather dependent. In fact, growth and development of crops under a particular set of climate parameters defines geographical location, productivity and production period of different crops. The magnitude of impact of climate and weather on agricultural productivity and quality of produce is appreciated by farmers and the scientific community, including horticulturists. Abiotic and biotic factors govern crop production potential and quality of products. Among the major constraints in production of horticultural crops are temperature (hot or cold), sunlight duration and quality, water deficiencies or excesses, atmospheric moisture (relative humidity), weeds, deficiency of nutrients, heavy winds, carbon dioxide and host of diseases and insect pests. There are ecological optima for obtaining production potential of each of the crops. Deviation from these conditions results in yield losses partially and sometimes totally. However, near optimal climatic conditions could be created by controlling the climate with the help of greenhouse using

The article is the Inaugural address delivered by Dr. R.S. Paroda at Indian Society for Protected Cultivation (ISPC-registered in March 2012), New Delhi on 21 March 2013 in a seminar ‘‘Advances on Protected Cultivation’’ at NASC, Complex, Pusa, New Delhi different protected structures/methods/devices and such cultivation under controlled environmental conditions is termed as as protected cultivation.

The Distributed Information Sub Center at Kerala Agricultural University, Thrissur was set up in May 1995 as the 23^rd center of the BTISNet of DBT, Government of India. The area of specilisation of the Centre is Agricultural Biotech- nology. The Centre passed through several phases involving Internet connectivity through the ordinary C band connectivity, then the Ku band connectivity etc. Now the commu- nication facilities of the Centre have been strengthened by procuring an ISDN Internet connection and a connection through ASIANET. The Center caters to the needs of the students (UG, M Sc & Ph D) and faculty of all departments of the Colleges in the main campus as well as of the other nearby academic and research centers of the University. Over the years, DISC has come to occupy a position in the KAU in the area of Infor- mation Technology , information sources and services based on Internet, CD-ROMs etc. The Centre has procured databases in the field of Biotechnology and developed several databases in the field of Agriculture and related fields. The center regularly conducts training programmes in Bioinformatics. The research activities in the field of Bioinformatics have been enhanced. The Centre has now been upgraded to DIC to facilitate intensive research on Bioinformatics in an effective manner. During the year 2004, the Centre has also started offering a course on Bioinformatics for the M.Sc Biotechnology students of the College of Horticulture.

Biotechnology research in KAU spread over various research stations and cam- puses includes Plant Biotechnology, Animal Biotechnology and Fisheries Biotechnol- ogy. Thrust areas for Plant Biotechnology include developing in vitro protocols for mass multiplication of elite types of fruit plants, ornamentals, medicinal plants, spices, plantation crops and forest species. The other thrust areas include checking clonal stability and genetic fidelity of in vitro derived plants using molecular markers, DNA finger printing and characterisation of germplasm with molecular markers, imparting biotic and abiotic stress tolerance in otherwise accepted crop varieties, genetic trans- formation for imparting/ improving expression of desired traits, in vitro germplasm conservation, enhancement of secondary metabolites production etc.

The crops identified for biotechnology research at KAU include spice crops like black pepper, ginger, turmeric, vanilla, cardamom; plantation crops like cashew, coco- nut and medicinal & aromatic plants such as Kaempferia, Trichopus, Gymnema, Holostemma and Coscinium. For improvement of such crops through molecular breed- ing, it is highly essential to identify/design specific probes/primers, micro-satellites, etc. By combining the research results generated at the Biotechnology Centre and informa- tion retrieved from the Bioinformatics Centre, suitable probes, primers are being de- signed for characterization and improvement of these unique crops through molecular breeding.

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The Cry2A proteins are promising candidates for managing the development of Cry1Ac resistance in insects. The Cry2A class of proteins is different from that of Cry1A, and experiments have shown that insects resistant to Cry1Ac are not cross-resistant to Cry2A proteins. However, the Cry2A proteins are less toxic than Cry1Ac to the polyphagous pest Helicoverpa armigera. Structure-based protein engineering of Cry toxins may pave the way for directed evolution of variants with broad spectrum of insect species specificity, enhanced potency, and stability. In the present study homology modeling of Cry2Ac was done using DeepView software. The template for Cry2Ac protein sequence (accession number X57252) was selected using ExPdb template search. The raw sequence of the Cry2Ac was superimposed over the Cry2Aa PDB template (1I5P). The superimposed model was submitted to the SWISS Model server. After receiving the Cry2Ac model, energy minimization was done using Gromos96. The threading energy and force field energy were calculated. The homology model of Cry2Ac made by us along with the Cry2Ac model from the SWISS-MODEL Repository was used for comparative analysis of receptor binding regions of Cry2Aa. Results showed variation between the two highly homologous Cry2A proteins in their receptor binding regions which may account for differences in spectrum of insecticidal activity.

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Astudy was undertaken at Department of Soil Science & Agrl. Chemistry, College of Horticulture, Vellanikkara to standardize different amelioration techniques to prepare enriched coirpith compost. Twelve treatments were tried using different combinations of coirpith, Pleurotus spp., Schizophyllus spp., urea, rock phosphate, cowdung slurry, mushroom waste, goat manure, glyricidia and water hyacinth. The results of the various enrichment techniques revealed that the temperature in the compost in general, increased to a peak value of 47oC on the 22nd day of composting and later decreased to a stabilized value. The observed temperature of 40-50 °C found during the thermophilic stage is essential for the rapid degradation of lignocellulose, as the thermophilic microfungi and actinomycetes involved in this process thrive at that temperature. Though the pH show a hike at the initial stage of composting, it decreased and stabilized at the later stage of composting which is conducive for growth and proliferation of microbes. The treatment coirpith with Pleurotus spp., glyricidia +20% cowdung slurry and goat manure was found to be ideal for compost preparation.

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Biological lipids are chemically diverse group of compounds. Lipids serve two major biological roles namely a structural one as component of membrane, and storage function, particularly in certain seed tissues. Fats and oils are the principal stored form of energy in many organism, and phospholipids and sterols makeup about half of the mass of biological membranes.

An experiment was conducted to study phospholipids composition qualitatively as well as quantitatively in the roots of mung bean seedlings. Among the four solvents used for phospholipids isolation, solvent II (methanol) and solvent III (chloroform: methanol: NH₃ salt) were proved to be the major phospholipids eluting solvents. The results indicated that Phosphatidyl Choline (PC), Phosphatidyl Inositol (PI), Phosphatidyl Serine (PS), Phosphatidyl Ethanolamine (PE) and Di Phospho Glyceride (DPG) were present in significant amount in the roots of ten days old mung bean seedlings. These phospholipids were also identified by their Rf values using thin layer chromatography. Among all the phospholipids isolated, PI showed the minimum Rf value, whereas PS showed the maximum Rf value. On the basis of column chromatography, four fractions of phospholipids were isolated. The highest amount of phospholipids was found in fraction II and the lowest in fraction I. Fraction II and fraction III contained the maximum phospholipids. In the fraction II PC showed the highest content (μg g-1) and one unidentified phospholipids showed the lowest content (μg g-1), whereas in fraction III, PI showed the maximum content (μg g-1) and PS showed the minimum content (μg g-1).

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The potato is one of the most important food crops both in developed as well as in developing countries. Developing countries today produce 37 per cent of world’s output of potatoes. Potato is an important food crop in India. Both area under potato cultivation and production has increased manifolds during past decades. The efforts are being made to improve potato quality and its post harvest life as well.

An experiment was conducted to isolate mitochondria from potato tubers and succinate dehydrogenase (SDH) assay and protein profiling were performed in the tissue. Three varieties namely, Kufri Bahar, Kufri Lauvkar and Kufri Jyoti were taken into consideration for the present investigation. Kufri Bahar showed the minimum values for SDH activity and Kufri Jyoti the maximum SDH activity. On the basis of SDH activity, the mitochondria in Kufri Jyoti were seemed to be more viable, functional and coupled as compared to Kufri Bahar and Kufri Lauvkar. This finding also suggested that the respiratory degradation of starch in Kufri Jyoti was higher as compared to Kufri Bahar and Kufri Lauvkar. Thus these result showed that the shelf life of Kufri Jyoti was less as compared to Kufri Bahar and Kufri Lauvkar. Protein profiling performed by SDS-PAGE was found to be similar in all the potato tubers.

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Apot culture experiment was conducted on tomato with cadmium and lead to identify the selective retention sites of the metals. For this a completely randomized design with five treatments and four replications were laid out at the College of Horticulture, Vellanikkara. The different levels of treatment ranged from 0.5 to 2.0 mg kg -1 soil. Cadmium and lead were directly supplied as a water-soluble source through cadmium chloride and lead nitrate respectively. In tomato the growth and fruiting was permitted only at lower levels of cadmium application. At 1.5 mg kg-1 soil, fruiting failed. While at 2.0 mg kg-1 soil, the very establishment of plant was totally affected. At higher levels of application, cadmium preferred to get accumulated in roots while on the contrary at lower levels, cadmium uptake increased in shoots than in roots with the least content of the metal in fruits. However, in the case of lead, all levels of application in soil permitted growth and fruiting. With increase in rate of lead application, significant selective retention was observed in roots (1.48-6.69 ppm), shoot (0.29-1.62 ppm) and fruits (0.39-1.45 ppm). On comparison of retention sites of lead in tomato, it could be seen that fruits maintained the maximum lead uptake (637.2-2933.0 mg pot-1). Hence, the consumption of such tomato fruits contaminated with heavy metals may result in kidney damage, lack of memory, dissolution of phosphorus in the bones etc.

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Detection of disease tolerance with enhanced defense mechanism is a practical approach in crop variety improvement programme. Toxins being responsible for pathogenicity and virulence of the fungi, regenerating plants from toxin treated calli is an effective and easy approach. The response of tomato calli to the crude toxins of the foliar pathogens viz., Alternaria solani and Septoria lycopersici was studied and the changes in phenol metabolism due to toxin treatment were analysed. Crude toxins of A. solani and S. lycopersici extracted under optimum conditions were serially diluted and tomato calli were treated with different dilutions of the toxins to find out the optimum dilution at which minimum survival percentage was obtained, based on a 1-4 score. Minimum score of 1 was obtained at 1:10 dilution for A. solani and at 1:20 for S. lycopersici.

The calli of resistant (LE-625) and susceptible (CO-3) tomato varieties were treated with desired dilutions of the toxins of A. solani and S. lycopersici. Total phenolics and ortho- dihydroxy (OD) phenolics were estimated at 24 h interval to study the extent of necrosis on resistant and susceptible calli due to toxin treatment. The total and OD-phenol contents of calli of susceptible cultivar (CO-3) progressively decreased with increase in time interval consequent to the treatment of toxins of A. solani and S. lycopersici. In the resistant calli of LE-625, an initial reduction of total phenol and OD-phenol content was observed following the application of the toxins of A. solani and S. lycopersici at 24 hours after treatment. Later with lapse of time, the total phenol content and OD-phenol contents progressively increased reaching a maximum at 120 h after treatment. The results clearly indicated the enhancement of biochemical defense in the resistant calli and a fall in defense mechanism in the susceptible calli due to toxin treatment. Such techniques are useful in the preliminary screening of resistant genotypes in crops like tomato.

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In Kerala, apart from cultivated sesame species (Sesamum indicum L.) three wild species were identified viz. Sesamum malabaricum, Sesamum mulayanum and Sesamum radiatum. An investigation was carried out to determine the intensity of arbuscular mycorrhizal fungal colonization in cultivated and wild species of sesame. The percentage of arbuscular mycorrhizal fungal infection in roots of different sesame species were found to be variable. The highest percentage of root colonization of arbuscular mycorrhial fungi was found in wild species S. malabaricum, which promotes tolerance to the plant under drought stress conditions.

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Atotal of 45 Gram positive, spore forming and rod shaped bacteria identified as Bacillus species were isolated from the rhizosphere of Chickpea plants. They were studied for their interaction with root borne fungal pathogens was studied using spot test method. Isolates CBS 29, CBS 113, CBS 123, CBS 132, CBS 153 and CBS 155 inhibited the growth of four or more fungi with inhibition zones varied from 1-5 mm. Two isolates CBS 153 and CBS 155 produced siderophores on CAS supplemented agar medium. Further inoculation of these Bacilli on seeds of Chickpea cultivar C235 and H8618 showed inhibitory, stunting or stimulatory effects on plumule and radicle growth depending upon cultivar, bacterium used and stage of seedling growth. Studies on co-inoculation of these isolates with Mesorhizobium and their plant growth promoting activity are being carried out. These co-inoculation studies are important areas of exploitation to improve crop productivity.

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Black pepper (Piper nigrum L.), known as the ‘King of Spices’ is an important export- oriented spice crop, valued for its medicinal and condimental properties. A perfect protocol for the rapid in vitro multiplication of this plant is available. However, systemic bacterial contamination is the single major constraint in the large scale in vitro production of plantlets. Based on morphological and biochemical characters, this bacterium was identified as Beijerinckia indica It grows profusely on nitrogen-free Jensen’s agar and solubilizes CaCO in the medium by acid production. It is resistant to a range of antibiotics and produces indole acetic acid. The bacterium is slow growing and has pH optima of 5.5 and temperature optima of 27oC. It has both endophytic as well as epiphytic association with the plant, the colonization being more in the stem and petiole. A single plasmid of 4.0 kb size was isolated from the bacterium and characterized. Competent cells of E. coli DH5a, transformed with this plasmid survived on Jensen’s nitrogen free agar, indicating its ability to fix atmospheric nitrogen. We stipulate that the nif genes are located on the plasmid of the bacterium. Since endophytic bacteria are known to stimulate plant growth through antagonistic activity against pathogens, production of growth promoting substances and inducing systemic resistance in several other agricultural crop species, the symbiotic association between Bejerinckia indica and black pepper could be further exploited through integrated nutrient management for improved growth and yield of this spice crop.

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Trichoderma spp. are important group of fungi which are mycoparasitic on several plant pathogenic fungi. Trichoderma spp. are, therefore, used as commercial biofungicides. Genetic manipulation through induced mutagenesis and molecular cloning are two important tools in improving the biocontrol potential of Trichoderma spp. In the present study, attempts were made to induce genetic variability in Trichoderma virens using UV- induced mutagenesis. In the process, a simple protocol was developed for obtaining high frequency mutations in Trichoderma virens. Using a UV-crosslinker, four stable mutants of T. virens have been developed, which are distinctly different from the wild type in appearance and in antagonism against Rhizoctonia solani and Sclerotium rolfsii. One of the mutants, designated as TI is very slow in growth, but sporulates densely, and retained its mycoparasitic potential against R. solani. Though all the mutants were able to antagonize R. solani, all of them were overgrown by the test pathogen S. rolfsii. These mutants could be used to identify genes involved in growth and conidiation in T. virens.

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Soil samples from pepper growing tracts of six districts of Kerala viz., Ernakulam, Idukki, Kollam, Kottayam, Thrissur and Pathanamthitta were assessed for total microbial population. Thirty two fungi, 17 bacteria and three actinomycetes were isolated and studied for their antagonistic potential against the foot rot pathogen, Phytophthora capsici. All the 32 fungal isolates were found antagonistic to the pathogen and majority of them belonged to the genera Trichoderma, Aspergillus, Penicillium, Fusarium, Pythium, Cunninghamella and Rhizopus. Among these maximum inhibition was recorded by Trichoderma spp. which included T. pseudokoningii, T. longibrachiatum, T. polysporum and T. harzianum. The laboratory studies were followed by field trials laid out in farmers’ fields (newly planted pepper gardens) in six districts of Kerala. Effect of biocontrol agents in wilt incidence revealed that the two-time treatment of combined application of Trichoderma local isolate and potassium phosphate was superior in checking the foot rot incidence. The treatment, Trichoderma local isolate + Bordeaux mixture applied twice came in the second position. Data on biometric observations revealed a significant increase in the number of laterals in plots treated with Trichoderma local antagonist and potassium phosphate as compared to plots treated with Bordeaux mixture and copper oxychloride. Results of quality analysis of berries revealed that oleoresin content ranged from 6.98 - 11.84 per cent and piperine content from 3.61 - 5.92 per cent and the quality remain unchanged due to various treatments. The study has provided an ‘eco and health friendly’ biological control for the management of Phytophthora foot rot, the most devastating disease of pepper.

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An in vitro evaluation was carried out to find out the efficiency of bacterial and fungal antagonists in checking the growth of pathogens of Kacholam. Out of the two methods tried to find out the effectiveness of Pseudomonas fluorescens against the two isolates of Ralstonia solanacearum, cross streaking produced lysis at the juncture and point inoculation produced clear inhibition zones. Among the two fungal antagonists tested Trichoderma viride was more efficient than Aspergillus niger because of its faster growth over the pathogen. However, both the antagonists showed complete overgrowth and dense sporulation on the area where the pathogen was streaked. To find out the efficacy of Pseudomonas fluorescens against leaf spot pathogens like Colletotrichum gloeosporioides and C. capsici, streaking on both sides yielded more than 50 per cent reduction in growth of both the fungal pathogens. The fungal antagonist T. viride showed a faster growth over pathogens than A. niger eventhough both recorded an antagonism index of 1500. Dense sporulation was seen over the pathogens in case of T. viride. The ability of fungal antagonists to produce volatiles and their effect on fungal pathogens were also assessed. It was noticed that the volatiles produced by T. viride was more effective than that of A. niger. Even though the extent of inhibition was less compared to direct action, this could also be one of the mechanisms of action of both the antagonists. The study revealed that all the three antagonists were effective against both bacterial and fungal pathogens and they could be utilized under field conditions.

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The antagonistic organisms are known to bring out action on the pathogen by various mechanisms. Based on in vitro screening of epiphytic microbes from cocoa pod surface against Phytophthora palmivora, the incitant of pod rot of cocoa, a promising isolate of Trichoderma viride and two isolates of Pseudomonas fluorescens were selected. The mechanisms of antagonistic action of these selected epiphytic antagonists against P. palmivora were studied in comparison with the standard cultures of T. harzianum and P. fluorescens. The selected epiphytic fungal antagonist T. viride and standard culture of T. harzianum were found to be efficient parasites of P. palmivora. The hyphae of the pathogen were seen tightly held by coiling with hyphae of the fungal antagonists. The hyphae of the antagonists penetrated the host hyphae at several points and grew to its inner cavity. The fungal antagonists also caused disintegration of the pathogen hyphae, besides free intermingling, which resulted in malformation of host hyphae. The ability of the two epiphytic P. fluorescens isolates and standard culture of P. fluorescens in producing HCN and siderophores was assessed. It was found that these isolates produced HCN and siderophores in appreciable levels thereby suppressing the P. palmivora. Though, more than one mechanism may operate in suppressing the pathogen, the relative importance of a particular mechanism may vary with different conditions, which needed to be further investigated.

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An In vitro study was conducted to evaluate the fungicidal efficiency of four selected fungal antagonists against leaf spot pathogens of ivy gourd by dual culture method. The antagonists evaluated were Trichoderma viride, Trichoderma harzianum, Aspergillus niger and Chaetomium globosum. The major leaf spot pathogens of ivy gourd used for the study were Cercospora cocciniae, Colletotrichum gloeosporioides and Alternaria alternata, causing severe infection on the foliage. Among the four antagonists tested, T. viride and T. harzianum were found to be the most effective recording the highest AI value of 1500 against all the three pathogens followed by A. niger. C. globosum was least inhibitory on C. gloeosporioides and A. alternata, but was effective against C. cocciniae. A complete overgrowth of the antagonists on all these pathogens were observed in the case of T. viride, T. harzianum and A.niger. C. globosum showed aversion to the hyphal growth of C. gloeosporioides and A. alternata, whereas in C. cocciniae, a homogenous growth was observed.

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Interaction of cationic dye acridine orange with lipopolysaccharide (LPS) isolated from Rhizobium strains specific (IC59) and non-specific (TAL1000) to chickpea (Cicer arietinum.L L.) has been investigated by spectrophotometric measurements. The strain specific (IC59) lipopolysaccharide induced decrease in absorbance of the α-band of acridine orange at 490 nm and finally upon further addition of LPS /Dye ratio in the range of 30~60 giving rise to α-polymeric metachromatic band at 660 nm. In contrast the LPS from non-specific strain (TAL1000) induces a decrease in absorbance at the α-band till LPS / Dye ratios are 0~50. This suggest that the anionic changes of strain specific LPS is different from non-specific strain enabling formation of large acridine orange aggregate on the surface of specific strain.

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Phytophthora pod rot (PPR) incited by Phytophthora palmivora is the most serious disease of cocoa, inflicting heavy crop losses. Since, antagonistic microbes are useful in ecofriendly management of plant diseases, the present study was undertaken to harness the potential of antagonistic epiphytic microflora isolated from cocoa pods for the management of PPR of cocoa. A quantitative estimation of the epiphytic microflora from pods collected from different locations of cocoa gardens of Thrissur district revealed more abundance of bacteria followed by fungi, actinomycetes and fluorescent pseudomonads. The in vitro antagonistic effect of 16 isolates of fungi, 22 isolates of bacteria, five isolates of actinomycetes and two isolates of fluorescent pseudomonads from pod surface against P. palmivora was carried out in comparison with standard cultures of Trichoderma harzianum and Pseudomonas fluorescens. The study revealed that all the epiphytic fungal and bacterial isolates including standard cultures of T. harzianum and P. fluorescens were antagonistic to P. palmivora in varying degrees. Based on the in vitro screening seven promising epiphytic fungal isolates and five bacterial isolates along with standard cultures of T. harzianum and P. fluorescens were evaluated for their efficiency in reducing the infection by P. palmivora on detached pods. The pathogen was inoculated on the pods one hour after application of antagonists. Observations taken 10 days after inoculation of the pathogen revealed that one isolate of T. viride and two isolates of P. fluorescens obtained from pod surface exerted more than 40 per cent efficiency over control in reducing the infection. The standard cultures of fungal and bacterial antagonists recorded 33-35 per cent efficiency. Hence, these efficient epiphytic and standard cultures could be utilized for the management of PPR of cocoa.

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To study the antagonistic potential of rhizobacteria against the soil borne pathogen Rhizoctonia solani, soil bacteria were isolated from the rice fields of Agricultural Research Station, Mannuthy and screened against R. solani. Out of the 22 rhizosphere bacteria, 10 were found to be antagonistic to R. solani. Seven belonged to the genera Bacillus, Pseudomonas, Clostridium and Sporosarcina and the rest three being unidentified. The bacterial isolates viz., 2B, 4B, 19B (Bacillus sp.), 28B and 10B (Sporosarcina sp.) showed increase in inhibition on extending the period of incubation. Maximum inhibition was expressed by 2B on the third day of inoculation followed by 4B, 10B (Sporosarcina sp.), 7B (Bacillus sp.) and Pseudomonas fluorescens. The bacterial isolate 4B and 7B (Bacillus sp.) and 10B (Sporosarcina sp.) completely prevented the sclerotial production of R. solani whereas 1B (Clostridium sp.), 2B, 19B (Bacillus sp.) and P. fluorescens delayed the sclerotial production. Pot culture studies also confirmed the superiority of these rhizobacteria in controlling the infection by R. solani. The potentiality of these rhizobacterial antagonists can be exploited for the management of soil borne pathogens like R. solani.

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Banana is one of the most accepted and versatile fruit crops across the globe and in Kerala, it is the most important economically profitable fruit crop grown, mainly by small and marginal farmers, providing a vital source of income. However, the productivity of this crop is very much reduced due to various diseases of which rhizome rot disease is an important one. In recent years, this disease has become a serious problem to the commercial cultivation of banana in southern states of India like Kerala, Karnataka, and Tamil Nadu.In Kerala the disease incidence is found to be nearly 30% and it will soon become cent percent if unnoticed since the disease is more seen in banana cultivated in paddy fields, and the area under the cultivation of banana in paddy fields in Kerala is rapidly increasing. The disease is caused by a gram negative bacterium, Erwinia carotovora. Developing an integrated management package involving biocontrol methods is the only remedy for managing the disease within the financial limits of the marginal farmers. With this intension a pot culture experiment was conducted to study the inhibitory effect of different biocontrol agents and botanicals in the incidence of the disease after testing their efficacy under in vitro cond itions. The results revealed that the biocont rol agent Pseudomonas flourescens (commercial preparation @ 7g /plant) was found to effective in managing the disease. The co- efficient of infection was found to be13.9% as against the control pot where it was 56.24%. With regard to the growth parameters also, the treatment with this biocontrol agent gave superior results. Thus it was proved that Pseudomonas flourescens (commercial preparation @ 7g /plant) could be successfully utilized in the integrated disease management making it cost effective.