eChapters

ABSTRACT

The discovery of polymerase chain reaction method of amplification of DNA is an important milestone in molecular genetics and used for multiple purpose such as construction of linkage maps, marker saturation of specific genome regions, analysis of genetic diversity, molecular phylogeny and cultivar identifications (Williams et al; 1990, Demeke, Adams and Chibbar; 1992, Welsh and Maclelland; 1990;, Obara-Okeyo and Kako; 1998, Sawhoon Lin et al; 1998 and Grunger et al; 1998.). 15 species of three morphologically distinct groups of Orchids belonging to the subfamily Epidendroideae and tribe Dendroebieae and Cymbideae were studied using random amplified polymorphic DNA (RAPD) technique. Genetic variability and relation between these 3 genera and 15 species were estimated based on band sharing and cluster analysis. A total of 227 distinct major RAPD bands of which 97% were polymorphic were generated from 15 arbitrary primers. The number of bands generated per primer ranged from 3 (Primer A-10) to 22 (Primer K-10 and N-17) with a mean of 14.8 bands per primer. Out of 227 bands generated 222 bands (97.8%) where polymorphic while only 5 bands were monomorphic. The primer resolving power was found to range between 2.00 (prime A-10) and 13.38 (primer k-10) when all the species were studied were taken into account. The molecular analysis grouped all the species into 5 groups. The polymorphic patterns generated by RAPD profiles showed different degrees of genetic relationship among the species studied and the RAPD marker were found to be an useful tool for detecting genetic variation within the species of three important genera of Orchids.

ABSTRACT

Genetic variability was assessed among twenty five distinct drumstick (Moringa oleifera Lam.) landraces using PCR based random amplified polymorphic DNA (RAPD) markers. Genomic DNA extracted from tender leaves was used for RAPD analysis. Thirty five primers were screened and based on the number of bands three primers were selected. The primers produced 34 bands, of which 58 % (20 bands) were polymorphic. The data were analysed using NTSYS pc 2.1 software. The similarity matrix constructed using Jaccard’s coefficient method was subjected to cluster analysis using UPGMA (Unweighted Pair Group Method with Arithmetic Mean) and dendrogram was generated. The pair wise coefficient values in the genetic similarity matrix among the landraces varied between 0.28 and 1.00. The dendrogram based on the matrix separated the landraces with distinct morphological characters and it was noted that the cluster formation was not fully in agreement with the geographical location. The study showed the use of RAPD markers in detecting genetic variability present among the drumstick landraces studied.

ABSTRACT

There is a wide variability in the contents of the commercially important chemical compounds of Centella asiatica depending on the source / ecotypes of the plants. The glycosides found in Indian plants include brahmoside, brahminoside and minor amounts of asiaticosides, which vary in their relative contents depending on the ecotypes. An RAPD marker assisted study of ten accessions, collected from different geographical locations of Kerala, viz., Parumala (Kottayam District), Kottayam, Nagercoil (Tamil Nadu), Mananthala (Trivandrum District), Vlathankara (Trivandrum District), Ponmudi (High range, Trivandrum District), Angamaly (Ernakulam District), Punalur (Kollam district), Pattambi (Palakkad district) and Calicut, was conducted. DNA amplification of the accessions was studied using forty decamer primers, involving 43 cycles. Based on the number of bands four primers were selected. The least similarity coefficient of 0.71 was shown by the Pattambi accession, which formed a separate cluster with all other accessions. Nagercoil and Ponmudi accessions showed the highest similarity coefficient of 0.88 with each other.

ABSTRACT

In oil palm there are three fruit forms viz. dura, tenera and pisifera. The commercial planting material is tenera produced by crossing dura and pisifera (D X P). Dura has thick shell and less economic part, where as pisifera is an abortic, female sterile type and cannot be reproduced in the normal ways. When tenera is selfed, segregation takes place in the monogenic Mendelian ratio; 1 dura: 2 tenera: 1 pisifera. Since all these palm types are morphologically alike, they can be confirmed only after fruiting, and verifying the internal fruit morphology. Fruiting in pisifera is highly erratic and often prolonged to several years. Proper technique in identifying these palms has been puzzling the breeder all over. Hence, a shell thickness marker is very important for early identification of the oil palm varieties.

An attempt has been taken to screen PCR primers and select the probable primers linked to the shell thickness in the present study. ‘Bulk segregation analysis’ approach was adopted for the study and DNA isolation was carried out from 25 dura and 25 pisifera palms. All the palms were developed from same tenera x tenera crosses. DNA from all the 25 duras and 25 pisiferas were pooled separately. Two pooled DNA samples were subjected to RAPD analysis with 50 random primers using the standard RAPD protocol. The amplified samples were electrophoresed and documented.

Both the pooled samples showed same banding pattern with all the primers, except 10 primers showing differences. Since the genetic configuration of the pooled dura DNA was likely to be same as that of the pisifera pooled DNA, the primers showing difference in banding pattern were the putative primers linked to shell thickness. All the 10 primers would be used for amplification of individual dura and pisifera DNA for confirmation of shell thickness marker.

ABSTRACT

Of the several PCR techniques developed during the last two decades, random amplified polymorphic DNA (RAPD) offers a simple and economical means of genotype characterization. The information obtained through RAPD characterization is extensively used for the identification of genotypes, screening of duplicates, assessing genetic diversity and monitoring the genetic stability of conserved germplasm. The present study is a report on the DNA fingerprinting of P. nigrum and P. longum cultivars. Fourteen land races and three advanced cultivars of P. nigrum and eleven land races and one advanced cultivar of P.longum were used in the present study. Forty decamer primers from Operon Tech were screened using two representative genomic DNA samples of black pepper. Of these, 10 primers that yielded clear and dominant band patterns were selected for the final analysis of the 29 accessions. These generated 119 amplification products. The total number of markers per primer varied with respect to both P. nigrum and P. longum. Cultivar specific single bands were obtained for a few land races and accessions of both the Piper species.

ABSTRACT

Production of cardamom in India has been greatly affected by Cardamom mosaic virus (CdMV). The conventional breeding program to develop resistance is a lengthy process. An alternative approach is to develop transgenic cardamom through introduction and expression of viral sequences in the plant. The coat protein (CP) gene of all the CdMV isolates have been cloned in pGEMT vector and sequenced. Sequence comparison studies of the eight isolates of CdMV in their CP and 3’UTR showed that, Kursupara isolate is highly variable from the rest of the isolates (Jacob et al., 2003). Hence the present study was undertaken to introduce the Kursupara CP gene of CdMV into cardamom plants in order to develop pathogen-derived resistance in cardamom. The CP gene of the Kursupara isolate of the virus was amplified with and without the 3’untranslated region (3’UTR) from the viral clone. These PCR products were cloned into the pXcmkn12 vector and sub cloned in to a plant expression vector (pAHC17) at the BamHI site, under maize ubiquitin promoter. Work is in progress towards the biolistic transformation of cardamom with these constructs.

ABSTRACT

Zingiber officinale is a herbaceous perennial. The absence of seed set in ginger makes conventional breeding methods inapplicable. Genetic modification through biotechnological means is the only way for introducing novel genes into the cultivated types. Transient expression of the gus reporter gene was used in preliminary experiments. Agrobacteruim tumefaciens strains EHA105/p35SGUS Int. effective in expressing b- glucuronidase activity was used for standardizing the optimum conditions for effecting transformation, such as preculture of explants, bacterial dilution, infection time, co-cultivation period etc. In addition, effects of acetosyringone and explant wounding were evaluated using the same strain. Genetic transformation of bud and regenerated plantlets was confirmed by their ability to grow on MS medium containing kanamycin 100 mg l-1 + cefotaxime 300 mg l-1 by histochemical b-glucuronidase assays.

ABSTRACT

Coleus forskohlii Briq. is a threatened medicinal plant , susceptible to fungal disease. The regeneration of this plant has been achieved on MSo medium supplemented with 2 mg/l or 3 mg/l BAP and 0.2 mg/l 2,4-D for callus induction using leaf as explants. Multiple shoot production from the leaf-derived transformed callus of coleus has been standardized using the various concentration of NAA with 2 mg/l BAP. The combination of 2 mg/l BAP and 0.5 mg/l NAA was found to be best for multiple shoot production. Rooting was found to be best on MS medium supplemented with 0.7 mg/l NAA. Coleus forskohlii was transformed with glucanase-chitinase gene along with npt-II gene. The presence of npt-II genes in the leaves of putative transformants were confirmed by PCR analysis. SDS- PAGE analysis and glucanase assay revealed the over expression of glucanase enzyme in the transgenic plants. The transgenic coleus plants survived the infection caused by Fusarium chlamydosporium due to the accumulation of glucanase-chitinase enzyme, where as control plants were susceptible to the disease.

ABSTRACT

Fruit rot and die back caused by Colletotrichum capsici is the destructive disease in chillies. The potential of Pseudomonas fluorescens for the management of fruit rot of chillies was evaluated under greenhouse and field conditions. Among the various strains of P. fluorescens tested in vitro, the strain Pf1 significantly inhibtied the mycelial growth of C. capsici. Combined application of talc-based powder formulation of P. fluorescens to seed (10 g/kg seed) and foliage (1 kg/ha) effectively controlled the fruit rot disease. Early and high induction of various defense compounds were observed due to P. fluorescens treatment.

ABSTRACT

Chickpea is an important pulse crop of India and most suited to the use of rhizobia for its nitrogen requirement. Mesorhizobium ciceri which interact with the crop are poorly understood. Attempt has been made to select M. ciceri with several beneficial properties. Sixty four isolates of M. ciceri collected from the nodules of several chickpea varieties and mutant lines were screened for their growth properties on nitrogen free medium. Isolates MC59 and MC 18-7 showed normal growth and had ARA values at half level to that of Azotobacter. PCR amplification results suggested the presence of ACC deaminase gene sequence both in Azotobacter and M. ciceri. Invariably all isolates showed the presence of one plasmid in the range of 35 kb to 40 kb size.

ABSTRACT

Bacterial wilt caused by Ralstonia solanacearum is the major constraint for cultivation of tomato in Kerala. Biocontrol of plant pathogens is becoming an important component of integrated disease management. Among the various antagonists tested against R. solanacearum of tomato, fungi were more effective than bacterial and actinomycetes organisms. Among them, Trichoderma virens, T. viride (Ozhalapathy and Vellanikkara) and T. pseudokoningii were the effective ones, recording the maximum antagonism index (AI) value of 6000 under in vitro condition. They inhibited the pathogen completely by the mechanism of both lysis and overgrowth. In pot culture study, T. viride and T. pseudokoningii and commercial Pseudomonas fluorescens gave maximum protection against wilt incidence. Under field condition, T. viride (Ozh), T. pseudokoningii and P. aeruginosa showed the lowest wilt incidence of 50.0, 58.3 and 58.3 respectively against cent per cent incidence in the control plot.

ABSTRACT

Anursery experiment was conducted to study the effect of solarization, selected native antagonists viz., T. viride and T. longibrachiatum and the standard culture of T. harzianum for assesseing the growth enhancement and management of Phytophthora rot in black pepper nursery. The experiment was laid out at College of Horticulture with 20 different treatments and replicated thrice. The sprouting percentage of cuttings in different treatments was worked out at different intervals, which showed significant difference. In general, it was observed that the maximum sprouting percentage was noticed in treatments where solarized potting mixture and biocontrol agents were used. The population of soil microflora viz., fungi, bacteria and actinomycetes in different treatments were also estimated. The study indicated a fluctuation in the population of soil microbes in various treatments at different periods of observation. The observations on height and number of leaves of pepper cuttings in different treatments were recorded. It was noticed that cuttings raised in solarized potting mixture incorporated with native antagonists, especially T. viride, exerted a significant effect in increasing the height of cuttings. With regard to total number of leaves, it was also observed that addition of native antagonists in solarized potting mixture had a positive effect in increasing the leaf production. Thus, the effect of solarization enhanced the growth parameters in crop plants by making available the required nutrients. The increased growth response of plants by soil application of Trichoderma spp. may be due to the production of growth promoting substances by the antagonists. Thus, the nursery experiment revealed that, in general, soil solarization and incorporation of antagonists had a favourable effect in promoting the growth of pepper plants besides checking the Phytophthora rot in black pepper nursery.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

ABSTRACT

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.

INTRODUCTION

It can be added that physiological stress as well as the physical offences contribute as primary factor for disease occurrence and fish mortality. Many parasitic diseases due to fluctuation in water quality leads to deplition of stock, and some times total stock will go eg. yellow grub disease.

EFFECT OF TEMPERATURE ON PARASITES

Heterophyid Trematode

Cyprinodon variegatus, an estuarine fish, is heavily parasitized in the bulbus arteriosus by the heterophyid trematode Ascocotyle pachycystis. Parasitized fish ventricles were hypertrophied due to mechanical blockage of normal blood flow by metacercariae in the bulbus Swimming performance generally was reduced by the presence of parasites, which had their most significant negative effect at cold temperatures and low dissolved oxygen level ( Felicia C. Coleman , 1993)

Soil consistency is a term used to designate the manifestation of different physical forces acting within the soil at various moisture levels. The consistency limits, also known as ‘Atterberg Limits’ (Atterberg, 1911; 12) are defined by the water content of the soil to create specified degree of consistency. These limits are – (i) Liquid limit or upper plastic limit, (ii) Plastic limit or lower plastic limit and (iii) Sticky limit.

7.1. Determination of Liquid Limit or Upper Plastic Limit

Equipments and Materials

Liquid limit device (Casagrande, 1932) and accessories, spatula, dish, oven, moisture box, balance, desiccator

8.1. Law of Mass Action

Gulberg and Waage, the two Norwegian chemists in the year 1864 stated the law of mass action in this form: ‘The velocity of any chemical reaction at any instant at a given temperature is proportional to the active mass of each of the reactants at that instant present in the system, the active mass in a homogeneous system is defined as the number of gram molecule of the substance present per unit volume i.e., the molar volume’.

Mathematical Formulation of the Law of Mass Action

Let us take a simple case of a reversible reaction at constant temperature represented by the equation:

19.1. Plant Available Fraction of Nickel, Cadmium, Lead and Chromium

The DTPA method was originally developed to extract Mn, Cu, Fe and Zn from slightly acidic to alkaline soils (Lindsay and Norvell, 1978), but in some cases it can be used for other metals also such as Nickel (Ni), Cadmium (Cd), Lead (Pb) and Chromium (Cr). Among them Ni is considered as an essential micronutrient. The DTPA is used to extract soil metals by forming chelates with free metals in solution, which lowers their ionic activities so that additional quantities of metal are released from the soil until equilibrium is attained. Triehanolamine (TEA) is used to buffer the extracting solution at pH 7.3 in an attempt to optimize the extraction of various metals. Calcium chloride is included in the extracting solution to minimize dissolution of CaCO₃.

9.0. Introduction

Enzymes are organic biocatalysts which initiate and control biological reactions. A catalyst is an agent affecting the velocity of a chemical reaction without appearing among the final products of the reaction. Unlike general catalysts, enzymes are specific in nature and function at a temperature of approximately 37^o C and pH close to neutrality. All enzymes are proteins but all proteins are not enzymes. These are distributed among the cell and tissues of plants and animals. Use of malted barley in brewing, yeast in the production of alcoholic beverages and wrapping of meat in the bruised leaves of papaya tree for tenderizing meat are techniques that have been used for centuries. In bread and cheese making also, enzymes have been exploited from generation to generation. Thus, enzymes have been used without knowing the nature of enzyme as catalysts and reactions involved. However, it is only recently that nature and action of enzyme is known and its importance in food industry is realized.

Sprayer is a machine to apply fluids in the form of droplets. Sprayer is used for the following purpose.

• Application of herbicides to remove weeds.
• Application of fungicides to minimize fungus diseases.
• Application of insecticides to control insect pests.
• Application of micro nutrients on the plants.

The factors affecting sustainability of agriculture under small farm production system in coastal ecosystem of Odisha was studied to ascertain the status of ecological factors and suggest means and ways to improve the situation. The response of the sample farmers and the findings of the study together led to suggest the following measures for up-gradation of ecosystem of the coastal area leading to sustainability of agriculture. Like other states, Odisha is also facing with problems of sustainability of agriculture. Our sample districts namely Puri, Khurda, Ganjam and Balasore which are situated in coastal eco-system of the state has different problems concerning to natural resources like soil, water, forest and animal wealth, etc.

The suggestions to improve the sustainability status of agriculture have been viewed from two important angles. One is adoption of technological measures to preserve natural resources and other is the policy and extension approach to educate people about the necessity of taking such measures for sustainability of agriculture and improve upon.

The study entitled “sustainability of small farms in coastal ecosystem” was conceived out of consideration of continuous degradation of natural resources- base consisting of soil, water, forest, maintain and animal wealth of the state. With broad objective of investigation into the status of sustainability of agriculture in the state, the specific analysis like socio-economic and farming related situations, perception about sustainability of agriculture in terms of ecological soundness, economic viability, social accessibility, humaneness and adaptability, development of sustainable agriculture index, adoption of sustainable measures, extension components and suggestions and strategy to maintain sustainability were taken into consideration. The organic farming and agri-business opportunities for sustainable agriculture were also studied.

The study was conducted in the state of Odisha covering the districts of Puri, Khurda, Ganjam and Balasore. Altogether four blocks each of one district and eight villages two from each block were selected for the purpose of the study. As much as 200 respondents who were within definition of small and marginal farmers were selected for interview. The respondents randomly so selected were personally interviewed by the author through an interview schedule developed on the basis of objectives stated earlier. The secured data were processed statistically to reveal the relevant findings as per the need of the study. From the relevant findings of the study the conclusions have been summarized below.

• The term Spice and Condiments applies to natural plant or vegetable products or mixtures in whole or ground form, which are used for imparting flavour, aroma and piquancy to the food items.

• Under the act of Parliament, a total of 52 spices are brought under the purview of Spices Board.

• However 109 spices are notified in the ISO list.

• India is the largest producer, consumer and exporter of spices and spice products in the world and produces more than 50 spices.

• The spices production in India is of the order of 5.74 million MT from an area of about 3.07 million ha.

• Chilli is the major spice crop occupying about 25.8 percent of area under cultivation and contributing about 22.7 percent of total spices production in the country.

• Garlic accounts for 21.9% of production and 8.1% of area Turmeric accounts for 16.9% of production and 6.3% of area.

• Rajasthan occupies major area under spices owing to seed spices cultivation in the state followed by Gujarat, Andhra Pradesh, Madhya Pradesh and Karnataka.

• Andhra Pradesh, which is the foremost state producing chilli and turmeric in the country, ranks first in terms of production.

The term plantation crops refer to those crops which are cultivated on an extensive scale in contiguous area, owned and managed by an individual or a company.

There are two groups of plantation crops namely coconut, arecanut, oil palm, cocoa, cashewnut dealt by Ministry of Agriculture and tea, coffee, rubber dealt by the Ministry of Commerce.

These are high value commercial crops of greater economic importance and play a vital role in improving Indian economy, especially in view of their export potential, employment generation and poverty alleviation particularly in rural sector.

Landscaping is both science and art and is an aspect of the environmental impact of a hotel that needs to be addressed. Understanding and blending the elements of both nature and construction is effective landscaping. Whether Landscaping is extensive or small consumes a large amount of water, fertilizers and herbicides and generates solid waste.

Ecogreen’s Greenscaping Operation provides cost-efficient and environmentally friendly solutions for landscaping. Greenscaping operation deals with two aspects:

1. Solid Waste Management, and

2. Water Conservation

Trees are major capital assets in cities Just as streets, sidewalks, public buildings and recreational facilities are a part of a community’s infrastructure, so are publicly owned trees. Trees — and, collectively, the urban forest — are important assets that require care and maintenance the same as other public property. Trees are on the job 24 hours every day working for all of us to improve our environment and quality of life. Urban forest provides many environmental benefits to our community. Aside from the obvious aesthetic benefits, trees within our urban forest improve our air, protect our water, save energy, and improve economic sustainability.

In other words, since most trees have been planted much of the tree cover is in urban areas as opposed to “natural lands”. Therefore, estimated pollutant uptake rates are higher for residential compared to natural or unmanaged lands. Possible management implications of these estimates are that air pollutant uptake benefits from tree planting may be optimized by planting in areas where air pollutant concentrations are elevated and where relatively high planting densities can be achieved thereby enhancing the health of urban dwellers.

Soil testing kit is developed to determine the available nutrient status of soil by rapid test methods. With this system of soil testing, a farmer can test his own soil as often and, in as many places as he feels necessary, and after gauging the need of his soil, he can apply the fertilizers as required. The present mode of soil testing in laboratories usually takes few days and sometime weeks and the farmers may have to travel long distances to submit the soil sample.

The stability of yield is an important characteristic to be considered when judging the value of a cropping systems when compared to others cropping systems. Here, we focus on multiple cropping and how this agroecological practice can be used to increase the sustainability of crop production. Strip, mixed and relay intercropping can be used to increase crop yields through resource partitioning and facilitation. In addition to achieving greater productivity, diversifying cropping systems through the use of strategic cropping systems will increase yield stability, reduce weeds, pests, disease and improve soil health. Several multiple cropping systems are already implemented in industrialized agricultural landscapes, including mixed intercropping with perennial grasses and legumes as forage and relay intercropping with winter wheat and red clover. Because multiple cropping systems will provide numerous benefits, researchers should be clear about their objectives and use appropriate methods so as not to draw spurious conclusions when studying multiple cropping systems. In order to advance the practice, experiments that test the effects of multiple cropping systems should use standardized methodology and researchers should report a set of common criteria to facilitate cross-study comparisons. Multiple cropping systems with two or more crops appears to be less common with annuals than perennials, which is likely due to differences in the mechanisms responsible for complementarity. One area where intercropping with annuals in industrialized agricultural landscapes has advanced is with cover crops, where private, public and governmental organizations have harmonized efforts to increase the adoption of cover crop mixtures.

Introduction

Anatomy of the immune system in birds differs from that in mammals. Immune cells are present in primary lymphoid organs (PLO) and secondary lymphoid organs (SLO). In birds, thymus and bursa of Fabricius (cloacal bursa) are primary lymphoid organs. Secondary lymphoid organs include the spleen, bone marrow, Harderian gland, aggregates of lymphoid tissue in various organs; i.e. in the gut called gut-associated lymphoid tissue (GALT); in bronchi called bronchial- associated lymphoid tissue (BALT). SLO contains aggregates of lymphocytes and antigen presenting cells. Birds do not have lymph nodes.

In PLO, the lymphoid cells are processed for differentiation and maturation. T lymphocytes differentiate and mature in the thymus, whereas B lymphocytes mature in bursa of Fabricius. Functional lymphocytes move from PLO to SLO. Like mammals, birds defend the body against pathogens through innate immunity (non-adaptive immunity) and adaptive or acquired immunity. Under innate immunity system, physical barriers present in birds play their important role preventing the entry of pathogens into the body. Physical barriers are feathers, skin, aggregates of lymphoid cells in various organs and ciliary defence in the respiratory system. Phagocytosis is first defence for pathogens entering the body, evading the physical barriers. Phagocytic cells include mainly heterophils and macrophages. Natural killer cells (NK cells), thrombocytes and complement also help in innate immunity.

Adaptive immunity is a particular defence which develops when the innate immunity is inadequate to impart protection. Adaptive immunity is antigen specific. T cell, B cell and macrophages play a major role in adaptive immunity. Adaptive immunity is brought about by antibodies (humoral immunity) or also by cells (cell-mediated).

Chlamydial organisms are spherical 0.2-1.5 um in diameter. They can be stained and seen in the light microscope, but they grow only in living cells. Chlamydial organisms infect mammals and birds causing various disease syndromes. Chlamydiacae family has one genus; Chlamydia. C. trachomatis, C. suis and C. muridarum. They cause disease in man, swine and mice. C. psittaci, C. pneumonia, C. felis, C. Caviae, and C. abortus. cause disease in man, birds and animals. Species can be differentiated on their growth characters inside the host cell.

ND is a viral disease of poultry, wild and cage birds characterized by marked variation in morbidity, mortality, signs and lesions. Reported in 235 species of birds.

A disease of primary importance and concern worldwide which causes continuous massive economic loss. It is a reportable disease in many countries of the world.

Occurrence

Species: All birds susceptible. Temporary conjunctivitis may develop in people who come in close contact with NDV for the first time.

IB is an acute, highly contagious, viral disease of chickens characterized by respiratory signs (gasping, sneezing, coughing, and nasal discharge), the severe renal disease associated with neurotropic strains, and a marked decrease in egg production. It is a serious cause of sub-optimal egg production and poor egg quality. This disease is worldwide in ditribution.

Hosts

Chickens only. Recently IBV has been detected in species other than chickens. The disease may be produced only in chickens, IBV may multiply in other species of birds and be a source of infection.

Etiology

IB is caused by the avian coronavirus; numerous strains of infectious bronchitis virus (IBV) exist because of high genetic diversity and cross protection between strains is not reliable.

IBD is an acute, contagious, viral disease of young chickens characterized by diarrhoea, vent pecking, trembling, incoordination, inflammation followed by atrophy of the bursa of Fabricius (BF) and by a variable degree of immunosuppression.

In the USA the disease was reported in Gumboro, Delaware in 1962, hence the name. The occurrence is worldwide.

LL is a retrovirus-causes, neoplastic disease of semi-mature or mature chickens characterized by a gradual onset in a flock, persistent low mortality and neoplasia of, the bursa of Fabricius (BF) with metastasis to many other internal organs especially the liver, spleen and kidney.

Vitamin A

Source of Vitamin

Berseem, legumes, alfalfa or other grass meal, fish oils, yellow corn, palm oil, commercial Vitamin A concentrate

6.1 Kinetics of Rumen Digestion

The rate and extent of digestion of dry matter or organic matter in the rumen is regarded as the important parameter to predict voluntary feed intake (Khazaal et al., 1993; Blummel,1994). Voluntary feed intake is responsible for more than 50 per cent of the variations in animal response to the diet (Van Soest, 1994) and with low quality unsupplemented roughages, this can be as much as 93% (Blumme1,1994). Therefore, any parameter of feed evaluation that would indicate the voluntary feed intake would be a useful parameter to judge the quality of feedstuff. The rate of digestion of any feed component can be measured by determining extent of digestion at intervals of time. Therefore, the techniques for determining extent of digestion, such as the rumen in vitro techniques (Tilley and Terry, 1963; Goering and Van Soest, 1970) or the in situ Dacron bag technique have been adopted to derive rate of degradation of dry matter of any particular component of the feedstuff. However, unlike in determining the extent of digestion, wherein the incubation is turned off at one particular time (normally 48 hours), measurement of rate of digestion, requires termination of incubation at several points between the start of incubation and until at least 90% of the potentially digestible dry matter is digested (72 or 96 hours). This generates a large number of samples for analysis.

In gas production technique, since gas produced from digestion of substrate is accumulated in the incubations, measurement of gas at different times of incubation would reflect the extent of digestion at different times of incubation. Therefore, unlike with other in vitro / in situ techniques wherein, digestion is determined from the gravimetric disappearance of feed at different times of incubation, gas production technique offers the possibility of using cumulative gas production at different times of incubation to measure the rate of digestion. Thus, a single incubation of feedstuffs is adequate to measure the rate of gas production.

Chromatography is one of the most effective technique for accurate, fast and convenient separation and identification of the components of a mixture. The method was developed by Russian botanist Mikhail Tswett (1906), described by Karl Runge (1855). Martin and Synage (1944) received Noble Prize for developing methodology of partition Chromatography.

Partition chromatography

Partition chromatogaphy is a system comprising of two physically distinguished components - a mobile component or phase and a stationary component or phase. A mixture of substances which are to be separated is placed on the system and separation takes place on the basis of difference in distribution of molecular species in two phases. Relative movement is a result of driving force and retarding effects.

Stationary phase is the sorbent, which may be liquid or paper or gel. If it is liquid then it is held stationary by adsorbing on a solid support or matrix. The mobile phase may be solvent or developer or a gas. Since the two phases are immiscible, the partitioning is defined as the distribution of solute in two immisicible phases which are in contact with one another. When a column is filled with the sorbent and solvent the two phases form a large number of theoretical plates. Each plate contains both the phases. Solutes in solution (mixture of substances to be separated) is layered on the top of the column and when it enters to the column it distributes in the two phases. The ratio of the concentration of a solute in two phases is called its partition coefficient. The resolution of solutes improves as the length of stationary phase increases.

Cotton (Gossypium) seed meal (CSM), the by-product of cotton seed oil industry, is widely used as an animal feed. However, its utilization as an animal feed is limited due to the presence of gossypol, a polyphenolie binaphthyl aldyde, which is toxic to non-ruminants such as poultry and swine and young ruminants such as calves and lambs. The gossypol (free gossypol, FG) present primarily in the pigment glands of cotton seed, is mostly converted to bound gossypol due to condensation of aldehyde group with free amino groups of proteins to form a schiff base. Formation of schiff base though results in the detoxification of gossypol to some extent, it also lowers the nutritive value by reducing the availability of lysine, the limiting amino acid of cotton seed. Thus, bound gossypol, consequently total gossypol (TG) (free plus bound) content is an important factor of cotton seed meal.