eChapters

The term DNA sequencing refers to methods for determining the order of the nucleotide bases, adenine, guanine, thymine and cytosine, in a molecule of DNA. In molecular terms ,the definitive understanding of a DNA molecule comes from determining its nucleotide sequence.The function of a gene can be deduced from its nucleotide sequence,for example by comparing the sequences of genes with known functions.

The first DNA sequences were obtained by academic researchers, using laborious methods based on 2 – dimensional chromatography in the early 1970s.  Following the development of dye-based sequencing methods with automated analysis, DNA sequencing has become easier and orders of magnitude faster.  Knowledge of DNA sequences of genes and other parts of the genome of organisms have become indispensable for basic research studying biological processes, as well as in applied field like diagnostic or forensic research.  The advent of DNA sequencing has significantly accelerated biological research and discovery.

1. Preparation of diluents 

The diluent is prepared one hour before semen collection so that it is stabilized. The diluent is prepared by mixing the ingredients and kept ready in water bath at 30oC. One of the diluents is detailed below

Participatory curriculum reorientation awareness-raising workshop for key stakeholders: In the present study focus group discussion and workshops were conducted to know the effectiveness of present curriculum of extension education with different stakeholders (Students UG, PG and Ph.D, Extension teachers, Field extension workers and farmers). Data were collected by using schedules regarding the important contents of present curriculum and most important for extension workers in the present situation to be included in curriculum

The study entitled “SHG in Techno Economic Empowerment of Tribal Women” was conducted on the following specific objectives.

Empowerment of women in relation to SHG (ii) Profile of SHG

Structure and functions of SHG

SWOT analysis of SHG

Constraints of SHG and suggestion

Case studies

Collection trips were organized in a planned manner so as to cover every locality in all three seasons. Rural and remote areas which were thus far practically unexplored were surveyed more intensively. Frequency of collection trips was twice a week during first year; once a fortnight in third and once a month in the last and forth years. During the field work, observations on the habit and habitat; flower colour; fragrance; fruiting, colour of the bark and blaze in case of trees and shrubs; dominance and sociability of the collected plants were entered into rough field book. In the mean time information regarding medicinal or other uses, if any, were gathered from local inhabitants, vaidyas and haquims. Plants of small size were uprooted carefully and pressed as such. In case of larger plants, care was taken so that specimens should have flowers, fruits, leaves and stems of young and old stages. At least four specimens, for making herbarium sheets, were pressed at spot after tagging; and a tagged bundle of a few plants was taken in a polythene bag for study in laboratory.

Research methodology is a detailed plan of investigation and the blue print of procedure for carrying out the research. In this chapter, discussion on the methodology has been made to understand the concepts, methods and techniques, which are utilized to design the study, collect information, analyzing data and interpreting the findings for revelation of truth and formulation of theories. The entire chapter has been broken up under following sub-heads for easy understanding:

1. Locale of research
2. Sampling design
3. Pilot study
4. Variables and their measurements
5. Methods of data collection
6. Statistical tools used for analysis of data

1. Introduction

The study of systematics and genetic diversity between or within different species, populations, and individuals is a central task for many disciplines of biological sciences. Systematic studies can indicate which genomes in the plant kingdom to search, sample, and study for the answers to questions relating to the evolution of chemical and physical structures and their synthesis or ontogeny as well as questions about the kinship of the individuals. Such studies would help find new and useful genes and provide information about the phylogenetic relationships among crop species and their wild relatives. The application of molecular markers has been the most active area of research in both animal and plant systematics. The analysis of the genetic variability within and among populations of the species is crucial for understanding their future maintenance and developing improvement as well as conservation programs.

Classical phenotypic characters, such as morphological traits, are still extremely useful but they can sometimes be influenced by environmental conditions. Therefore, during the past decade, classical strategies of evaluating genetic variability such as comparative anatomy, morphology, embryology and physiology have been increasingly complemented by molecular techniques. These include the analysis of chemical constituents (e.g., plant secondary metabolites) and, most importantly, the characterization of macromolecules. The development of molecular markers, which are based on polymorphisms found in proteins or DNA, has greatly facilitated research in a variety of disciplines such as taxonomy, phylogeny, ecology, genetics, and plant breeding (Weising et al. 1995). The greater utility of molecular markers derives from following five inherent properties that distinguish them from morphological markers.

1. Introduction

The objective of algal taxonomic collections is to represent the natural population, flora and diversity in size and form. Therefore, it is important to collect the representative samples seasonally from various habitats. Collection of information about the habitats and other details are also very important. For algal sampling the first thing to consider is how to collect the specimens. The freshwater algae show an ability to tolerate a wide range of environmental condition. Under natural conditions, they usually grow in mixed communities which may include many species and genera. Field study provides information about the site, environment, season and size and shape of the living algal thallus in its natural surroundings. Many algae can be tentatively identified by their appearance, shape and colour of thallus in a particular habitat. Algal population can be sampled in a number of different ways, depending on the nature of the habitat and the location of the algae within the water body. The specimens must be prepared carefully so that important morphological characters are displayed as fully and completely for further researches.

The methodology has been carefully considered in order to comprehend the idea, procedures, and approaches used in the design of the study, information gathering, data analysis, and finding interpretation for the identification of relevant truths and the development of hypotheses. The methodology and a process employed in the study are covered in this chapter.

5.1 Locale of the Research

The rural areas of Darjeeling-Pulbazar CD block, Jorebunglow-SukhiaPokhri CD block in Darjeeling district in West Bengal was selected for the study. The area had been selected for the study because of the availability of appropriate respondents, people as well as the local language.

5.2 Pilot Study

Prior to beginning the actual research, a pilot study was carried out to better understand the regions, its inhabitants, institutions, and programmes activities associated to the migration process in the research area. During the pilot study period, the respondents’ situational and historical data were gathered.

1. Introduction

The word ‘lichen’ has a Greek origin, which was referred to the superficial growth of fungus like organism on the bark of olive trees. Theophrastus, the Father of Botany coined the term ‘lichen’ during 300 BC and introduced this group of plants to the scientific world.

Lichen is a combination of two organisms, an alga and a fungus, living together in symbiotic association. Sometimes instead of an alga a cyanobacterium (blue-green alga) may be present in the lichen thallus. The algal component in the lichen is generally called as ‘photobiont’ as it contains photosynthetic pigment. The fungal part is called as ‘mycobiont’ (myco = fungus). The photobiont and the mycobiont loose their original identity in symbiotic association and the resulting organism is the lichen which behaves as a single organism, both morphologically and physiologically. Hence, the lichen is called as a composite organism. The lichen thallus is made 90% of mycobiont which provides shape, structure and colour to the lichen. In a lichen thallus whatever visible from outside is fungal part, which holds algae inside its body (Fig. 1) and therefore, the lichens are placed within the Kingdom - Fungi. The fungi present in lichens are called as ‘lichenized fungi’. Among the 20,000 lichen species present in the world, 98.9% of them belongs to Ascomycetes group while Bacidiomycetes and Deuteriomycetes groups are represented by only 0.1% and 1%, respectively.

Introduction

Age determination, a fundamental discipline within fisheries biology and ecology, is a critical tool for unraveling the life histories and population dynamics of aquatic species. The ability to accurately estimate the age of individual fish holds immense value in various scientific, management, and conservation contexts.

Seed Production

Systemized crop production is known as seed production. In seed production adequate care is given from the purchase of seeds upto harvest adopting proper seed and crop management techniques. The benefits of seed production are

• Higher income
• Higher quality seed for next sowing

Before carry out any ethnobotanical work, first define the goal of the assignment or study before selecting the approach, which best suits the interests, budget and schedule. Most of the methods used in ethnobotanical studies are time taking and costly because several field trips are required in several seasons for covering different ritual occasions and seasonal farming activities which are continue for a number of years for completion of an assignment. In this way, an assignment may be very long or short as per selection approach of an ethnobotanist. Long-term assignment or projects may be continue for several years and they require sufficient financial input. When sufficient financial add and time does not available to accomplished long term assignments, in that case a Rapid Ethnobotanical Appraisal (REA) can be done.

1. Introduction

Microbial diversity is becoming increasingly essential with the requirement to preserve the integrity, function and long-term sustainability of natural and managed ecosystems. However, our knowledge of the diversity of microbes remains obscure due to lack of appropriate approaches to evaluate the contribution of different components of the soil microbes to ecosystem. This lack of approaches together with the complex and heterogeneous matrix that soil microbes are embedded in, poses great challenges to understand the dynamics of microbial communities in soils (Nautiyal et al. 2010; Chauhan et al. 2013).

In classical terms, diversity is described as the total number of species present, i.e. species richness or species abundance, and the distribution of individuals among those species i.e. species evenness or species equitability. However, the incorporation of these two components into diversity indices has led to much controversy among ecologists (Atlas 1984). The Shannon-Weaver index (Shannon and Weaver 1949) is probably the most widely used index for measuring diversity of a microbial community. But this index, as many others that have been proposed, has its own limitations. A major difficulty is that both richness and evenness play a role in determining the value of the index. Therefore, any diversity index is a single value and thus cannot indicate the total make-up of a community. It is recommended that evenness, richness and diversity values should be calculated in order to obtain an objective assessment of community structure (Atlas 1984). From molecular point of view, diversity often refers to the number of different sequence type present in a habitat e.g. a soil (Ogram 2000). Measuring this type of diversity does not give information about the frequency of individual species, but it can give valuable information about changes in the community structure and species richness in response to changes in the physico-chemical properties of the soil and soil management practices (Nautiyal et al. 2010).

Solid formulations such as dust, wettable powder or granules, the active ingredient is mixed with inert material. The concentration is expressed as:

Active ingredient (%) in the total weight of commercial product.

I. Detection of Castor Husk in Oil Cakes and Oil Seed Meals (Method of ISI, (BIS), IS: 10165-1982)
Principle

The method is based on the fact that castor husk is not bleached under the conditions which cause the bleaching of almost all other materials of vegetable origin likely to be present in an oilcake. The method consists of the treatment of the material with dilute alkali and acid solutions followed by treatment with bleaching powder solution and the isolation of the unbleached castor husk.

Introduction

The main objective in analysing fertilizers is to assess their quality. The analysis examines both their physical and chemical composition. The quality of fertilizers is stated by the manufacturers and, in most countries, it is statutorily notified. Hence, analysis is carried out to determine whether the stated quality meets the statutorily notified standards or not. Together with the statutory notified composition of the fertilizers, the testing methods are specified. In situations where the testing methods are not notified, the prevalent standard methods are followed. Generally, the term fertilizer refers to mineral fertilizers, which are manufactured chemical products of standard composition, while the term manures refers to organic manures, compost, agro-industrial wastes, etc. The compositions of organic fertilizers, unlike mineral fertilizers, are quite variable and, thus, difficult to regulate precisely.

Fertilizer quality is notified in terms of physical and chemical characteristics. The physical parameters include moisture content and particle size. The chemical parameters refer to the amount and form of nutrients, and to various impurities that may be toxic to plants above a critical limit, e.g. biuret in urea. The efficiency of a fertilizer depends on its form of nutrient content. A phosphatic fertilizer may have water-soluble, citrate-soluble, water-insoluble or citrate-insoluble forms of phosphate. A nitrogenous fertilizer may contain ammoniacal, nitrate and amide forms of N in various proportions.

Straw represents a category of low-density feed characterized by a high concentration of structural carbohydrates, specifically cellulose and hemicellulose, while exhibiting low levels of carbohydrates, nitrogen and minerals, along with variable lignin content. Lignin serves as a physical barrier, inhibiting the microbial degradation of these materials. Straw is having low digestibility due to following reasons:
• Reduced voluntary intake due to unappealing taste.
• Low digestibility and nutritional value.
• Nutritional deficiencies, particularly in crude proteins, essential minerals, and vitamins.
• The presence of anti-nutritional components such as lignin, silica, tannins, and oxalates.
The primary aim of treating low-quality roughage includes:
• Enhancing voluntary intake by improving palatability.
• Increasing digestibility to boost the intake of digestible energy.
• Minimizing losses during digestive processes.
• Breaking down cellulose and hemicellulose from lignin and silica to enhance digestibility.
• Expanding the surface area available for microbial action and enzyme activity, thereby improving digestibility.
• Reducing bulkiness through densification.
The processing of low-quality roughages can be achieved through various methods.

Lipids are naturally occurring fat like substances insoluble in water but are soluble in organic solvents like alcohol, ether, and chloroform etc. Some of the phospholipids are even soluble in water. All the lipids do not resemble fats.
Lipids are an important component of living cells. Together with carbohydrates and proteins, lipids are the main constituents of animal cells. Lipids include fatty acids, neutral fats, waxes and steroids (like cortisone). Compound lipids (lipids complexed with another type of chemical compound) comprise the lipoproteins, glycolipids and phospholipids.
Properties of lipids
Lipids are non-polar molecules a family of organic compounds, composed of fats and oils yield high energy and are responsible for different functions within the body. It is stored in the adipose tissue of the body. Lipids are a heterogeneous group of compounds, mainly composed of hydrocarbon chains. Lipids are energy-rich organic molecules, which provide energy for different life processes. Lipids are a class of compounds distinguished by their insolubility in water and solubility in nonpolar solvents. It is important constituent of cell membrane in biological systems, a mechanical barrier that divides a cell from the external environment.
Most of the lipids or fats are demonstrated by using fat soluble dyes. The lipid soluble dyes have greater affinity for fats, lipids, lipoproteins and triglycerides. The fat soluble dyes are:

Introduction and Scope

Plant and soil testing enable scientific assessment of the needs of the plant for nutrient elements and of the capacity of the soil to supply them. The nutrient elements enter the plant in ionic form from the soil solution. Ion transport to the root surface may take place through ion diffusion and bulk transport (mass flow). Mass flow is the sweeping along of ions as water moves to the root. It is particularly important for ions that are not absorbed on soil colloids such as nitrates and sulphates. Because of mass flow, a plant deficient in N can extract all the nitrates from the soil. Potassium, phosphate and the micronutrient cations are absorbed on soil colloids with various degrees of affinity and are greatly retarded in movement with the soil water. Diffusion (movement along a concentration gradient) is the main mode of transport from the solid phase to the root surface for these non-mobile ions. There is also a possibility that some of them are absorbed by a direct exchange of an ion, usually hydrogen, between the root surface and ions absorbed on soil colloids. Most ions are sorbed by the root against a concentration gradient and, thus, the process involves the use of metabolic energy. The fact that nutrient uptake is an active process explains some of its peculiarities. Plants not only accumulate nutrients against a concentration gradient, they are also able to select from the nutrients at the root surface according to their requirements (preferential uptake).

The increasing global emphasis on sustainable and environmentally friendly energy alternatives has placed biofuels at the forefront of green energy innovation. Derived from agricultural biomass and organic waste, biofuels represent a promising substitute for conventional fossil fuels, particularly in agrarian economies where such feedstocks are abundantly available. Agricultural biomass includes crop residues such as rice straw, wheat husk, corn stover, sugarcane bagasse, and oilseed cakes. Meanwhile, organic waste comprises food waste, animal manure, municipal solid waste, and industrial organic residues. Converting these materials into biofuels not only reduces landfill burden and environmental pollution but also cuts down greenhouse gas emissions and promotes the development of rural and circular economies. A variety of biofuels—including biogas, bioethanol, biodiesel, biohydrogen, and syngas—can be produced through two major conversion pathways:
• Biochemical conversion, which involves microbial and enzymatic processes such as fermentation and anaerobic digestion.
• Thermochemical conversion, which includes pyrolysis, gasification, and combustion processes to transform biomass into energy-dense fuels.
Types of Liquid Biofuels
The term biofuel commonly refers to liquid fuels produced from recently living plant material, unlike fossil fuels derived from ancient biomass. Among these, ethanol and biodiesel are the most prevalent forms:
• Ethanol: A type of alcohol produced through fermentation, usually from sugar- or starch-based crops like corn, sugarcane, or cassava. Ethanol is used either as a fuel substitute or blended with gasoline to reduce carbon emissions in vehicles.
• Biodiesel: Produced through the transesterification of oils extracted from plants and seeds (such as soybean, sunflower, or palm), biodiesel is a renewable fuel that can be used in standard diesel engines. It offers a biodegradable, less polluting alternative to petroleum-based diesel.

6.1 Introduction

All separation procedures depend mainly on some physical characteristics of the compounds. It is relatively easy to separate compounds that have significantly different physical characteristics by simple techniques such as solvent extraction. If the various compounds are similar to each other in their molecular size, any slight difference in any one of their physical properties is exploited to achieve separation. The separation and purification of plant constituents is mainly carried out using one or other, or a combination of chromatographic techniques: Paper Chromatography (PC), Thin Layer Chromatography (TLC), Column Chromatography (CC), High Performance Liquid Chromatography (HPTLC), Optimum Performance Laminar Chromatography (OPLC), Gas Liquid Chromatography (GLC) and High Performance Liquid Chromatography (HPLC). One further technique, which has fairly wide application in phytochemistry, is electrophoresis.

All separation procedures depend mainly on some physical characteristics of the compounds. It is relatively easy to separate compounds that have significantly different physical characteristics by simple techniques such as solvent extraction. If the various compounds are similar to each other in their molecular size, any slight difference in any one of their physical properties is exploited to achieve separation. 

Introduction

Soil is the main source of nutrients for the plants. Soil also provides support for plant growth in various ways. Knowledge about soil health and its maintenance is critical to sustaining crop productivity. The health of soils can be assessed by the quality and stand of the crops grown on them. However, this is a general assessment made by the farmers. A scientific assessment is possible through detailed physical, chemical and biological analysis of the soils. Essential plant nutrients such as N, P, K, Ca, Mg and S are called macronutrients, while Fe, Zn, Cu, Mo, Mn, B and Cl are called micronutrients. It is necessary to assess the capacity of a soil to supply nutrients in order to supply the remaining amounts of needed plant nutrients (total crop requirement - soil supply). Thus, soil testing laboratories are considered nerve centres for nutrient management in crop production systems and plantation forestry.

Soils may have large amounts of nutrient reserves in them. All or a part of these reserves may not be of any use to crops because they may not be in plant-available form. For the purpose of estimation or analysis of plant-available soil nutrients, such methods are to be used that have been tested/verified for the correlation of nutrients extracted and their plant availability. This book describes internationally accepted and widely used methods. Apart from nutrients, soil pH estimation is also critical in the assessment of soil health. Generally, plants prefer soils that are close to either side of neutrality. However, there are acid- loving plants and also plants that can withstand high soil alkalinity. Hence, good yields and economic returns from farming are possible in acid and alkali soils. With proper amendments, still higher yields can be obtained in acid and alkali soils. Soil pH also has a considerable influence on the activity of soil micro-flora and on the availability of soil nutrients to crops. It is also important to estimate physical properties such as soil texture, structure, bulk density, soil colour, water content and soil temperature.

Abstract

Estimates of genetic parameters are needed for the prediction of breeding values and planning of selection strategies for desired genetic advancement. Even animal breeders feel lost when confronted with the huge set of different methods. This paper describes an overview of latest methods for the estimation of genetic parameters in a simplest possible way avoiding complex mathematical expressions.

Introduction

Irrigation water always contains some soluble salts irrespective of its source. The suitability of waters for a specific purpose depends on the types and amounts of dissolved salts. Some of the dissolved salts or other constituents may be useful for crops. However, the quality or suitability of waters for irrigation purposes is assessed in terms of the presence of undesirable constituents, and only in limited situations is irrigation water assessed as a source of plant nutrients. Some of the dissolved ions, such as NO₃, are useful for crops.

Sometimes, effluents and domestic sewage water is utilized for irrigating field crops. For using these water safely in the farming system, assessment of undesirable pollutants and organic and microbial constituents with regard to their suitability is fore-most important. Determination of organic constituents is generally carried out under two categories viz., (i) organic substances that quantify an aggregate amount of organic C and (ii) individual or specific organic substances, such as benzene, DDT, methane, phenol and endosulphan. Important determinations are the chemical oxygen demand (COD), which gives the total organic substances, and the biochemical oxygen demand (BOD), which gives the amount of total biodegradable organic substances in the water sample.

Organic material especially, seeds, grains, leaves, fruits and nuts contain prominently three groups of compounds. They are carbohydrates, proteins and fats/oils, mostly in that order. They also contain 3-5% minerals and smaller quantities of vitamins and other minor constituents. Cereal grains contain more of carbohydrates while oilseeds and nuts contain more oil. Tree nuts and even oilseeds may contain 30-50% protein while cereal grains contain 5-8% protein only. The starch and sugar contents of oilseeds may be less than 10% while the oil content of cereal grains ranges around 5%. The actual content and the composition of these groups of compounds really account for their quality as well as commercial value.

Seed is a basic input to raise the crop plants. Healthy seeds produces the healthy crop. However, the seed carries different pathogens either externally or sometimes internally in the seed to affect the seed emergence and healthy seedling growth. Therefore, the seed treatment with fungicides/bactericides, based on the infection of the pathogen, is highly essential to counter the attack and infection of these pathogens to the germinating seedlings.

Introduction

Diseases are a common occurrence on plants, often having a significant economic impact on yield and quality, thus managing diseases is an essential component of production for most crops. These fungicides are used because of three main reasons:

a.To control a disease during the establishment and development of a crop.

b.To increase productivity of a crop and to reduce blemishes.

c.To improve the storage life and quality of harvested plants and produce.

Application Methods

To protect the crop from fungal pathogens, fungicides are applied in various formulations (as dust, granules, gas and most commonly, liquid). They are applied to:

1.Seed treatment

2.Soil treatment

3.Protection of standing crop

Proper selection of a fungicide and its application at the correct dose and the proper time are highly essential for the management of plant diseases. The basic requirement of an application method is that it delivers the fungicide to the site where the active compound will prevent the fungus damaging the plant. The fungicidal application varies according to the nature of the host part diseased and nature of survival and spread of the pathogen. The method which are commonly adopted in the application of the fungicides are discussed

1. Foliar or vegetative application
2. Soil application

Proper selection of a fungicide and its application at the correct dose and the proper time are highly essential for the management of plant diseases. The basic requirement of an application method is that it delivers the fungicide to the site where the active compound will prevent the fungus damaging the plant. The fungicidal application varies according to the nature of the host part diseased and nature of survival and spread of the pathogen. The method which are commonly adopted in the application of the fungicides are discussed.

1.Soil treatment

2.Seed treatment

3.Foliar or vegetative application

The term ‘Ecology’ deals with the study of organisms from all sources and their interaction with surrounding environments. The aim of ecology is, therefore, to understand and also to explain the relationships of all organisms to their environment. In this respect, the duty of the ecologist is to study a particular environment, estimate the population of each species, recognize the communities, and determine their activities and their interactions with both related species, the rest of the community, and their environment. Macroecological studies always initiate by defining the species composition and then proceed to draw certain conclusions on the role of these species in the ecosystem. On the other hand microbial, especially bacterial classical taxonomy (based on isolation techniques and determination of morphology and in vitro biochemical characteristics of isolates) for determination of each species of aquatic bacteria is almost impossible, except in a few cases. Consequently, the microbial ecologist has to depend upon certain methods that can measure the biomass or activity of a very large conglomerate of coexisting microbial taxa in an aquatic environment. Probably, this has caused the very slow development of microbial ecology concepts and application of a general ecological theory to the microbial ecology.

1. Detection means the existence of whatever do you want detecting but diagnosis is that the analysis in depth of existing thing.
2. Objectively look for, listen to the credible complains, symptoms that is sometimes not visible at all.
3. It is art or skill of identifying some abnormalities, i.e. which is below or higher than the normal permissible range/threshold.

A. Evaluation of the efficacy and resistance of anthelmintics against Helminths Anthelmintic resistance According to WAAVP (World Association for the Advancement of Veterinary Parasitology) anthelmintic resistance is a failure to reduce feacal nematode egg count by at least 95%. Technically accurate definition is that resistance is a genetically determine decline in the efficacy of an anthelmintic against a population of parasite that is susceptible to the drug. The history of parasite resistance to anthelmintic starts with the first report on phenothiazine resistance in 1957. Haemonchus contortus was the first nematode to develop resistance against different anthelmintic. The World Association for the Advancement of Veterinary Parasitology has recommended two tests for detecting anthelmintic resistance in ruminants, horses and pigs. These are in vivo test (FECRT) and an in vitro test (the egg hatch test). Detection Technique

Objectives

1. To know the methods of essential oil extraction of spices and condiments.
2. To know the various chemicals present in the essential oil of different spices and condiments.

Essential oil: Essential oils are the liquids that are isolated from plants when introduced to solvents. They are liquefied versions of the plants.

Extraction methods of essential oil

1. Steam Distillation
2. Solvent Extraction
   1. CO₂ Extraction
   2. Maceration
   3. Enfleurage
3. Water Distillation
4. Water and Steam Distillation

Detection and identification of the causes and diagnosis of the diseases rapidly and precisely are the basic requirements for developing effective crop disease management systems suitable for different ecosystems. Incorrect diagnosis leads to failure to contain the disease spread and consequent additional expenditure to the growers. Hence, it is essential that proper diagnostic procedures are followed to achieve the goal of reducing both quantitative and qualitative losses of agricultural and horticultural produce. Morphological, biological, biochemical, physiological, immunological and genomic characteristics of microbial plant pathogens have been carefully considered to develop rapid, sensitive and reliable methods of detection and identification of cause(s) of newly observed disease (s) in the locations concerned. Various methods of establishing the nature of microbial plant pathogens are discussed in detail in earlier publications

2.1  Write True/False

Q.1    From theoretical point of view, the most general method of estimation is the “Maximum Likelihood Estimation Method.”
Q.2    The method of “maximum Likelihood Estimation” was initially formulated by the statistician named as “C.F. Gauss.”

4.1 Introduction

Extraction is a process whereby the desired constituents of a plant are removed using a solvent. The primary ways for extraction of organic molecules of interest to biologists and medical investigators involve breaking open the cells. Cell rupturing is carried out in a variety of ways. The method used depends on the type of tissue used. For plant cells grown in cell suspension culture or for plant callus tissue, a French press or a sonicator can be used to break open the cell. Plant cells grown in culture can also be ruptured with a glass tissue homogenizer. Silicified or highly lignified plant tissues within the organs such as roots, stems, leaves, fruits and seeds are usually frozen and pulverized using liquid nitrogen in a mortar and a pestle (Kaufman et al., 1999). Softer tissues can be ground in a small volume of buffer in a mortar, washed white sand and a pestle to rupture the cells.

When the cells have been ruptured, the extraction is performed using the appropriate techniques. Water soluble compounds and proteins are extracted in buffers or water. Organically soluble compounds are extracted with organic solvents. Boiling ethanol is a good all-purpose solvent for preliminary extraction (Harborne, 1998). The success of the extraction with alcohol is directly related to the extent chlorophyll (when isolating substances from green tissue) is removed into the solvent. The most common chemical procedure for obtaining organic constituents from dried plant tissues is to continuously extract powdered material in a Soxhlet apparatus with a range of solvents, starting with petroleum ether and chloroform (to separate lipids and terpenoids) and then using alcohol and ethyl acetate (for more polar compounds). This method is useful when working on the gram scale. However, one rarely achieves complete separation of constituents and the same compounds may be recovered (in varying proportions) in several fractions. The extract obtained is clarified by filtration and is then concentrated in vacuo in a rotary evaporator. To prevent fungal growth, a trace of toluene may be added to the concentrated extracts and they should be stored in refrigerator. There are short cuts in extraction procedures which one learns with practice. The concentrated extract may deposit crystals on standing.

Extraction is a process whereby the desired constituents of a plant are removed using a solvent. The primary ways for extraction of organic molecules of interest to biologists and medical investigators involve breaking open the cells. Cell rupturing is carried out in a variety of ways. The method used depends on the type of tissue used. For plant cells grown in cell suspension culture or for plant callus tissue, a French press or a sonicator can be used to break open the cell.

Methods of fertilizer application

Two major methods are used to apply fertilizers  
I) Application of fertilizers in solid form 
II) Application of fertilizers in liquid forms

In order to get maximum benefit from manures and fertilizers, they should not only be applied in proper time and in right manner but any other aspects should also be given careful consideration. Different soils react differently with fertilizer application. Similarly, the N, P, K requirements of different crops are different and even for a single a crop the nutrient requirements are not the same at different stages of growth. The aspects that require consideration in fertilizer application are listed below:

Objective

1. To know the different types of fertilizer application methods applied in spices and condiments.

Methods of fertilizer application: The different methods of fertilizer application are as follows:

a) Broadcasting

1. It refers to spreading fertilizers uniformly all over the field.
2. Suitable for crops with dense stand, the plant roots permeate the whole volume of the soil, large doses of fertilizers are applied and insoluble phosphatic fertilizers such as rock phosphate are used.

Broadcasting of fertilizers is of two types

1. Broadcasting at sowing or planting (Basal application): The main objectives of broadcasting the fertilizers at sowing time are to uniformly distribute the fertilizer over the entire field and to mix it with soil.

Developments in recombinant DNA technology have made it possible to transfer gene from any source across species barrier. In fact, transfer and expression of foreign genes in plant cells is now a routine procedure. Of the several methods tried so far, the system based on Ti plasmid of Aprobacterium tumefaciens appears to be most successful for transformation of plant cells.

Dry Methods
1. Decortication and dehulling: The process of decortication for groundnuts and cottonseeds enhances the nutritional profile of the resulting cakes by eliminating the outer layers, which are notably high in crude fiber. Similarly, dehulling or dehusking of pulses and grains also contributes to an improved nutritional value.
2. Cracking and dry rolling: This technique involves the fragmentation of the kernel into smaller particles through the application of pressure using dry rollers. This method increases the surface area of the grains, thereby enhancing their digestibility.
3. Crimping: In this method, the traditional rolling of grains is substituted with the use of corrugated rollers, a process referred to as crimping.
4. Crumbles: The term “crumble” refers to the process of grinding pellets into a granular form, which is particularly beneficial for chick feed.
5. Popping or puffing: This technique involves the dry heating of grains, which must contain a moisture content of 10-14%. The final moisture content in the popped grains is approximately 3%. During this process, grains are subjected to temperatures ranging from 370 to 425 degrees Celsius for 15 to 20 seconds, leading to the rupture of the endosperm due to the rapid expansion of steam. This results in an increase in grain size and enhances both palatability and digestibility.
6. Micronizing: This process employs infrared heat energy on grains, resulting in the breakdown of glucosidic bonds between starch molecules.
7. Roasting: In roasting, grains are exposed to direct flames at a temperature of 148.9 degrees celsius. This method not only expands the grains but also inactivates protease inhibitors in whole soybeans, thereby improving their nutritional quality.

8.1 Introduction

After isolation and purification of the compound, it has to be identified. It is necessary to find out the class of the compound first and then to find out the specific compound within the class. The class of compound is usually determined from the colour tests, solubility, Rf properties and UV spectral characteristics of the compound. Further identification within the class depends on measuring physical properties and spectral measurements and then comparing these data with those in the literature. The physical properties include melting point (for solids), boiling point (for liquids), optical rotation (for optically active compounds) and Rf. The spectral measurements include Ultra Violet (UV) , Infra Red (IR) , Mass Spectrum (MS) and Nuclear Magnetic Resonance (NMR). Data on X – ray crystallography of the compound are also useful for identification of the compound.

On completion of the analysis of the spectra obtained, the pieces that have been identified are listed. Then the pieces, the MW and / or molecular formula are checked and the pieces are refined to fit. Finally, the pieces are assembled paying particular attention to ¹H-NMR chemical shifts and coupling patterns. The answer should be checked with the ¹H-NMR data very carefully and it is probably the most critical test to pass. The ¹H-NMR spectrum is potentially the most useful for assembling the structure of the compound.

A known compound can usually be identified on the basis of the above mentioned physical and spectral characteristics. For final confirmation, direct comparison with authentic compound (if available) should be carried out Authentic compounds can be obtained from commercial companies. In the absence of authentic compound, the data from literature can be carefully compared for the identification of the isolated compound. If a new compound is present, all the above data are sufficient enough to identify it. However, data on chemical degradation or laboratory synthesis of the compound are necessary to confirm the identification of the new compound.

After isolation and purification of the compound, it has to be identified. It is necessary to find out the class of the compound first and then to find out the specific compound within the class. The class of compound is usually determined from the colour tests, solubility, Rf properties and UV spectral characteristics of the compound. Further identification within the class depends on measuring physical properties and spectral measurements and then comparing these data with those in the literature.

13.1 Incubation
The silkworm eggs are to be incubated properly to avoid irregular hatching. An ideal incubation is to maintain a temperature of 25oC and 75-80% humidity continuously. In most of the places incubators are not available. In such cases, the eggs are kept spread in wooden trays provided with paraffin paper for base and the cover and foam pads kept wet all around inside the trays to maintain 70-80% humidity. In each chawki tray of 90 cm × 120 cm about 4 sheets of eggs can be kept for incubation. The room temperature is maintained at 25°C. On 9th day the egg sheets are rearranged. About 25 dfls (disease free layings) are kept in a chawki tray of 90 cm × 120 cm to enable brushing and proper spacing for young worms later. The egg sheets are covered with a sheet of black paper or paraffin paper. On the due date of hatching and when a few larvae have started hatching, black paper and the paraffin paper are removed and the eggs are exposed to dim light for about an hour. This facilitates the uniform hatching of larvae.
13.1.1 Methods of Incubation
There are two methods, i.e., constant temperature incubation and raised temperature incubation

The deliberation on the methodology has been made to understand the concept, methods and techniques which have been executed for delineating the study, collection of information, analysis of data, and interpretation of the findings for the revelation of hard evidences and formulation of theories. This chapter deals with the method and procedure used in the study consisting of eight main parts

A. Locale of research

B. Pilot study

C. Sampling Design

D. Item analysis and experts rating for devising scoring technique on dependent variables

E. Empirical measurement of the variables.

This chapter deals with an in-depth description of the methods and procedures employed in conducting the research in an organized manner. The research methodology gives the study’s work plan. The research objectives and theoretical base offer an initial framework for identifying the appropriate methodology to answer the research questions.

The procedures followed were described under the following headings.

5.1 Research design

5.2 Locale of the study

5.3 Content analysis of websites

5.4 Statistical tools used for the analysis of data.

5.5 Preparation of report

8.1 Introduction

After knowing when to irrigate and how much to irrigate, next important thing is how to irrigate the crop field. Methods of irrigation play a very crucial role in deciding how to irrigate the crop fields since all the crops do not require same methods of irrigation. Depending on the sources of water available for irrigation, type of crop to be irrigated, quantity and quality of water to be applied and other economic considerations, methods of irrigation will vary. Irrigation water may be applied to the crops by flooding it on the field surfaces as surface irrigation, by applying it beneath the soil surface as subirrigation, by spraying it under pressure as sprinkler or overhead irrigation or by applying it in drops around the root zone of the crop as trickle irrigation. The commonly used methods of irrigation are described in schematic diagram as follows: