eChapters

Communication is a key to understand things between two people. Today’s communication is entirely different from pre-historic era and has taken many evolution throughout the years. Before alphabets, signs, symbols, letters and telephone evolved, communication was restricted to interpersonal interaction (i.e.) from one person to another in the form of gestures. It is in other terms called non-verbal communication. Today, communicating between two or more people has become too easy that one can communicate with more than 10 members at a time which is in technical means called conferencing.

The growth of internet has paved way for innumerable means of communication. In other words it can be told as, communication is re-defined by technology. Texts, email, tweets, tele and video conferencing have widely saved much time of people and made them recipients of today’s fast moving world.

Communication beyond history

Ancient nomads used to communicate with the help of drawings and paintings. Chauvat Cave in France is one such example which proves that communication through cave painting had taken place 30,000 years ago Before Christ (BC). Throughout the world many such cave painting monuments has been discovered. One in South Sulawesi, Indonesia and Coliboaia Cave in Romania gives an idea about how communication would have prevailed among people those days.

Later came symbols. Historical evidences prove that, symbols were used by people around 10,000 B.C. Through rock carvings pictograms were developed by ancient people to tell a story to their next generations through logographic images. It is said that, culture among human groups have evolved through rock carvings. Later, ancient cultures developed ideograms. Egyptians had their own style of communication called Hieroglyphs. Chinese created their own characters. Lastly, the alphabets re-defined language and communication was developed around 2000 BC.

A laboratory is essential for research work in science and development. Most of the research institutes have dedicated laboratories for researchers and students to conduct experiments, perform research work, as well as to provide hands-on learning/training to students. There are certain rules and regulations laid down by the laboratory in-charge which need to be followed strictly for safety purposes. Some of the laboratories keep ‘safety manual’ for researchers to read before entering into the laboratory to conduct experiments. Whenever safety is compromised in a laboratory, there is increase in risk of unwanted incidents in the laboratory. Storing food/beverages in the same refrigerator where laboratory chemicals are kept may lead to disastrous effects on health. This chapter aims to provide instructions in brief on the things to be performed (Do’s) before and after entering onto a laboratory as well as the things never to be done (Don’ts) in a laboratory for the safety reasons.

Good Laboratory Practices (Do’s)

• P Wear apron/labcoat while working in the laboratory.
• P Use gloves/respiratory mask/fumehood while handling toxic/volatile/inflammable chemicals/reagents.
• P Use pippet or pippetor for safety in transfer of solutions.
• P Protect eyes, face and naked parts of the body from UV radiations. Use safety goggles or face mask/apron.
• P Keep your eyes away from Liquid Nitrogen and use cryogloves while handling it.
• P Arrange all the necessary materials/solutions before starting the experiment.
• P Periodically fumigate/ozonize culture and inoculation rooms.
• P Autoclave all contaminated materials before discarding.
• P Pre-treat gel or solution containing Ethidium Bromide (EtBr) before discarding into laboratory waste-disposal bin, and use gloves while handling them.
• P Burn all the transgenic plant materials in an incinerator after taking necessary observations.
• P Make entry in the designated log-book and clean the instrument after using it properly.
• P Switch-off the instrument after use or when not in use.
• P Wash hands with soap and remove the labcoat before leaving/going out of the laboratory7.
• P Ensure that switches for fans/lights, and water taps are off while leaving the laboratory.
• P Follow the biosafety guidelines suggested for handling and disposal of the hazardous chemical.

Introduction

Several hazardous chemicals, microbes, and techniques like recombinant DNA technology are used for research and development work in research Institutes and universities. Therefore, the researchers, technical, helping and supporting staffs as well as the laboratory outputs must be kept free from contaminations. Thus, biosafety is necessary to prevent unintentional exposure/ release of hazardous chemicals, microbes/pathogens, and toxins to the laboratory staff and environment. Following the biosafety guidelines and ethical considerations are essential to maintain safety standards for the researchers as well as the environment. This chapter deals with various biosafety measures and ethical issues to be strictly followed while working in a research laboratory. The required safety level needs to be determined as per the chemical/biological materials are used in the laboratory to keep the researcher/environment safe for sustainable research and developmental activities.

Biosafety

The term biosafety is used to describe the procedure and policies needed to ensure environmental and personal safety. Biosafety refers to the containment principles, technologies and practices that are implemented to prevent unintentional exposure to pathogens, toxins, or their accidental release into the environment. A fundamental objective of any biosafety program is the containment of potentially harmful chemical/biological agents as precautionary safety measures.

Why Biosafety?

With the increasing number of countries adopting molecular biology tools and techniques in life science research and development activities, especially in the areas of agriculture and medicine, the biosafety issues are gaining importance to ensure biological safety for the public and the environment. Conducting research in safe and sustainable manner is not only a personal requirement but essential collective efforts to ensure biological safety for a clean and safe environment. This requires rules, regulations, monitoring bodies, and awareness among the workers as well as the public. Therefore, biosafety per se is an integral part of laboratory research and requires awareness among the researchers so that biological safety can be well taken care from the grassroot level.

For a researcher, laboratory is another home where he/she spends much time every day. As we take care of cleanliness, safety, and design of our home, we must take care on these aspects of laboratory too. It is the responsibility of the laboratory personnel to ensure cleanliness/safety of the laboratory. Since every laboratory chemical and equipment have the potential to cause harms if not used with appropriate precautions. Hence, as a laboratory user, it is our responsibility to follow the basic safety/biosafety rules and regulations set up by the Principal Investigator of the laboratory, Institutional Biosafety Committee, as well as by other regulatory authorities, if any. We must be aware of the general/personal protective measures, cleanliness and laboratory waste disposal prior to joining a research laboratory. In this chapter we will briefly discuss about personal safety, chemical safety, cleanliness, waste disposal, etc.

A. Personal Safety

1. Personal Protective Equipment (PPE)

Personal protective equipment (PPE) is special arrangement to protect the worker from specific hazards because of using a hazardous substance. PPE does not reduce or eliminate the hazard, but protects the wearer from exposure to hazardous material. PPE should not be reused or brought outside the laboratory as these might increase the risk of spread hazardous material or contamination. Used PPE must be kept inside the laboratory in a designated waste storage area adjacent to the work area.

Introduction

Proper electrical wiring and earthing are crucial for laboratory safety and functionality. A laboratory houses several installations/equipment that demand stable/continuous power supply. Inappropriate or faulty wiring and earthing may lead to significant risks, including electrical shocks, damage to equipment, and even incidence of fire. Therefore, understanding the requirements of electrical wiring and earthing is essential for designing, construction, and maintenance of laboratory. This chapter aims to provide a basic knowledge of electrical wiring and earthing necessary for a laboratory. Importance of earthing in mitigating electrical hazards and ensuring integrity of electrical installations are also described.

Basics of Electrical Wiring

Basic understanding of electrical wiring is necessary for laboratory safety and efficient electrical functioning. The potential difference between two electrical points (negative and positive) is known as Voltage, which drives the flow of electric current. This is measured as volt (V). The rate of flow of electricity in a circuit is known as Current (I), which is measured as ampere (A). Current can be either direct (DC) or alternating (AC). Opposition/resistance experienced in the flow of current in circuit is known as Resistance (R) and it is measured in ohms (Ω). Higher the resistance means lesser the current flows at a given voltage. The rate at which electrical energy is transferred/utilized in an electric circuit is known as Power (P), which is measured in watt (W). The Power can be calculated as the product of voltage and current (P = V × I).

Introduction

We come across a variety of chemicals every day in a laboratory. While some of them are relatively safer, others are extremely hazardous. Thus, it is very important to know the potential hazards that can be caused by the chemicals present in the laboratory and the safety measures needed to be followed to avoid the hazards. Additionally, we should also be aware of the steps necessary to be taken on such hazardous situations. According to IUPAC, a hazardous chemical mean the chemical which has the potential to cause damage in various ways. In a laboratory, there are chemicals that might be irritant, carcinogen, respiratory fibrogen, systemic poison, asphyxiant, flammable, etc. which have the potential to cause hazard to the workers or environment. Therefore, it is important to use appropriate procedure to reduce the risk and protect the health/safety of laboratory personnel, the public and the environment.

Types of Hazards Associated with Chemicals

Chemical hazards: The hazards caused by chemicals can be broadly classified into three categories.

1. Health hazard: This is also known as toxic effects of chemical. The risk associated with a chemical in laboratory depends on the extent of exposure and the inherent toxicity of the chemical. Many chemical reactions form the product that might be much more toxic than the reacting chemicals. For example, inadvertent mixing of formaldehyde (a common tissue fixative) and hydrogen chloride results in the generation of bis(chloromethyl) ether, a potent human carcinogen. For most of the chemicals, the route of exposure (through skin, eyes, gastrointestinal tract, or respiratory tract) is more important (Figure 17.1) that must be considered during risk assessment.

Introduction

Flowering plants (Angiosperms) are identified by their morphological characteristics like flower, fruit and seeds that help their taxonomic characterization into different species, genera, families and orders. The floral characters which are used for taxonomic characterization of plants include the type of inflorescences, perianth structure, floral symmetry, arrangement of floral leaves, type of androecium, stamens, gynoecium, carpel, ovule, etc. The type of fruit and seed also play important roles in taxonomic classification. In this chapter we will briefly discuss about various morphological features of flowering plants (in botanical terms) that are useful in taxonomy.

Nomenclature of Plant

Nomenclature of plant is the formal and scientific naming of plants. It is related to but distinct from taxonomy for grouping and classifying plants and provide botanical name.

The scientific/botanical nomenclature started with Species Plantarum by Carolus Linnaeus in 1753. Botanical nomenclature is governed by the International Code of Nomenclature for algae, fungi, and plants (ICN), which replaces the International Code of Botanical Nomenclature (ICBN).

ICBN and ICN

International Code of Botanical Nomenclature, (ICBN) was adopted by the VIIth International Botanical Congress held in Stockholm, July1950. While the International Code of Nomenclature for algae, fungi, and plants (ICN) was adopted by the XVIIIth International Botanical Congress held in Melbourne, Australia in July 2011.

Introduction

Test of pollen viability (extent of pollen viability) is important in pollination experiments to ensure fertility/seed setting. It is also important in monitoring state of pollens during storage in ecological/ taxonomic studies as well as during research on pollen biochemistry, genetics, stigma interactions, and incompatibility studies. Pollen viability refers to the ability of pollen to accomplish post-pollination events to achieve fertilization. Pollen viability test has gained importance particularly in plant breeding and hybrid seed production experiments/ practices. In this chapter, you will be acquainted with the basics of pollen viability test. However, for more professional knowledge on the topic readers are suggested to refer specialized books.

Pollen Viability Test

It is important to know the extent of viability of pollens used in pollination experiment, in finding fertility of a plant of interest, to monitor state of pollen on storage, in ecological/taxonomic studies, as well as in research on biochemistry, genetics, stigma interaction, incompatibility assay, etc. of pollen. Several pollen viability tests have been developed time-to-time. It is recommended to initially use different tests for a given pollen type to find out the most appropriate method that provides reliable result on pollen viability. It is important to note that every procedure requires standardization of protocol according to the sample used. Following different tests are used for assessing pollen viability.

1. In-vitro Pollen Germination Test

It is a simple, speedy, and quantitative test for assessing pollen viability. In this method, a collection/sample of pollen is germinated in vitro and observed under microscope. The percentage of germinating pollens (producing pollen tube) after appropriate treatment and a given period of time is determined. The percentage provides an index of viability of the pollen sample.

Introduction

Seed is a living entity (mature ovule) consisting of a tiny plant in embryonic stage along with the stored food protected by an outer layer known as seed coat. Seed is an important biological input in agriculture, which is economically important for successful crop husbandry. Cotyledons serve as the source of nutrients (proteins, carbohydrates, and oils) that not only nurture the plant at the early developmental (seedling) stages of plant but also the vegetarian (heterotroph) organisms. Being one of the important inputs in agriculture, viability of seed needs to be assessed to know the germination percentage of seeds used to raise a crop. Since viability of seeds may deteriorate on storage depending on the storage conditions. Therefore, often it becomes important to test viability/germinability and vigour of seeds before being used for raising a crop or conducting an experiment. In this chapter, we briefly discuss about the different methods which can be used for testing the viability of seeds.

Seed Viability

Seed viability can be defined as the capability of the seed to germinate and produce a normal seedling within a specified period. In other words, seed viability indicates how many seeds in a lot are alive and could develop into plants under appropriate field conditions. It is very important that most of the stored seeds are capable of germinating when sown in a field. They must possess higher viability at the beginning of storage and maintain it over the storage. A seed with high initial viability survives longer in storage; however, the viability deteriorates slowly at first and then rapidly as the storage prolongs. It is important to know the duration of storage after which a considerable decline in viability occurs under given storage conditions so that to take necessary action to revive the seeds. Excessive deterioration in vianility of seeds may lead to loss of the seed/material/ germplasm.

Introduction

Tissue culture refers to the technique of growing cells, tissues, or organs under controlled, artificial conditions. This allows propagation of tissues from a small sample of cells/tissues under sterile conditions. Plant tissue culture is widely used for various purposes, including the production of disease-free plants, rapid multiplication of plants with desirable traits, genetic modification, conservation of rare or endangered species, etc. Tissue culture is a valuable technique because it enables consistent and large-scale production of plants, ensures genetic uniformity, and allows manipulation of plant biology at cellular level. In this chapter, we will discuss about the major types of tissue culture techniques and their advantages, disadvantages, as well as applications in brief.

Different Types of Tissue Culture

Callus culture involves induction and growing of unorganized, undifferentiated cell masses from an explant on a nutrient/culture medium. The part of plant used to induce callus is called explant which can be tissue, organ or part of a plant.

Organ culture is used to maintain or grow an entire organ or a part of organ in vitro. It preserves the structure and function of the organ for studies or transplantation.

Embryo culture involves isolating and growing embryos from seeds or ovules in a nutrient medium. It is used to bypass seed dormancy or rescue embryos from hybrid crosses that may not develop naturally.

Cell suspension culture involves growing dispersed cells or small cell clusters in a liquid medium under continuous shaking. This is used for large-scale production of cells, secondary metabolites, and studying cell biology..

Introduction

Different types of culture media are used for culturing microbes and plant tissues. Preparing as well as sterilizing them properly following appropriate method with safety for the desired purpose are important parts of research work. In this chapter, we will discuss briefly about different types of media, their composition, preparation and sterilization in laboratory for microbial or plant tissue culture.

Culture Medium

Ingredients mixed to prepare solid, liquid or semi-solid substance to support the growth of microorganism or plant tissue in vitro (outside the body) is called culture medium. Frequently, it becomes important to grow microorganism or plant/animal tissue in vitro for any or all of the following purposes:

• To study characteristic features or properties of microorganisms.

• To identify the causal organism(s) of infection in a clinical sample, so that appropriate remedy can be determined.

• To culture/multiply microorganism or plant/animal tissues in vitro for research or commercial purposes.

• To prepare/produce biological molecules, vaccines, antigens, proteins, metabolites, etc. in vitro for research or industrial purposes.

Introduction

Use of acid/base is common in laboratories for experiments like titration and proper safety in their handling laboratory is necessary. In this chapter we will briefly discuss about the safety measures necessary to be adopted while using acids/bases in laboratory for any experiment.

Handling of Strong Acid/Base

There are four major points that need to be taken care of while working with strong acids or bases that might be quite harmful if any accident happens.

1. Protect Yourself

Wear a lab-coat and acid-resistant gloves: Always use lab-coat/apron and make it sure that the sleeves properly cover your arms. More importantly, use gloves made of acid-resistant materials and also wear shoes to protect your feet while wearing long-pants/jeans. Female with long hair, must tie her hair properly (with rubber band) to avoid any disturbance caused by appearance in between.

Wear safety goggles: It is important to wear safety goggles to protect your eyes using larger and fitting goggles that cover front as well as sides of your eyes. Always use properly fitting and adjustable goggles because smaller/ loose goggles may not protect the areas around your eyes against accidental acid spills and it may fall down from your face, if it is too loose.

Locate emergency wash area: There must be an emergency washing area nearby the work place or laboratory containing lab spill management kits. All the workers in the laboratory must be aware of using the kits and emergency plans.

Neutralize acid: Don’t panic even if an acid or chemical spill happens. Stay calm and act as quickly as possible. In case an acid spills, neutralize it by using sodium bicarbonate so that they can be cleaned and disposed safely. If acid comes in contact of skin, wash it off under running tap water but do not use/ apply sodium bicarbonate on skin.

Introduction

Measurement of pH of a solution and preparation of different types/strength of buffers are common practices in a laboratory, particularly in the Biochemistry and Molecular Biology laboratories. Therefore, in this chapter you will be acquainted with the procedures of measuring pH of a solution as well as preparation of some of the commonly used buffers in a laboratory.

Potential or Power of Hydrogen Ion (pH)

Measurement of pH of a solution is measuring the molar concentration of hydrogen ion [H+] in the solution, which is an indication of the acidity or basicity of the solution. The symbol pH stands for “power of hydrogen” and numerical value for pH is just the negative logarithm of the molar concentration of hydrogen ions [H+]. The ionic product of water (Kw ) is the basis for the pH scale. The total hydrogen ion concentration from all the sources and it can be experimentally measured.

The pH scale designates the concentration of H+ (and thus of OH−) in an aqueous solution, which ranges between 1.0 M H+ and 1.0 M OH−. The term pH is defined by the expression

Thus, the ionic product of water makes it possible to calculate the concentration of H+, with given concentration of OH−, and vice versa. The value of 7 for pH of a neutral solution is not an arbitrarily chosen figure; it is derived from the absolute value of the ionic product of water. A solution having pH greater than 7 is basic, and the concentration of OH− is greater than that of H+. A solution having a pH less than 7 is acidic.

Introduction

For agricultural applications as well as for research work, often we need to prepare solutions of different agrochemicals of varying strength, as per the requirements. Agrochemicals are used to help crop plants grow, produce, and protect them from biotic/abiotic stresses. Many of the agrochemicals (insecticides, herbicides, fungicides, rodenticides, molluscicides, nematicides, etc.) are used to protect the crop from insect pests and diseases. However, some of them (fertilizers and soil conditioners) are used to augment crop yield. While in the previous chapter (Chapter 8) we discussed about preparation of laboratory solutions, in this chapter we will briefly discuss about the preparation of different agrochemicals.

Agrochemical/agrichemical is a generalized term used for the chemicals applied in agriculture. In general, it also includes the feed-supplements used in f isheries and animal husbandry.

Agrochemicals

Fertilizers

Fertilizers are used to augment growth of crop plants and to mitigate nutrient deficiency in the soil. Fertilisers can be organic (containing natural substances, prepared through natural processes) and inorganic fertilisers (synthetic fertilizers, manufactured artificially using chemical processes using natural deposits, by chemical alterations).

Soil Conditioners

Soil conditioners include manure, livestock and leaf compost, peat that help making the soil in good condition. They are laid on the top soil and then mixed properly. They not only increase nutrient contents in the soil but also improve aeration and water holding capacity.

Introduction

In a research laboratory, we frequently need weigh chemicals and prepare different solutions of varying strength, as per the requirements. Accuracy in weighing and preparation of solution, with respect to weight/volume and strength, determines the success of experiment. In turn, the accuracy depends on weighing of solid chemical and/or measuring the volume of liquid chemical/making up the volume. In this Chapter, weighing of chemicals using appropriate weighing balance, preparation of solutions of different strength, their dilution, and precautions while preparing will be discuss in brief.

Weighing of Chemical

The primary method to determine the amount of chemicals to be used is weighing the chemical by using a balance.

In general, there are two types of balances to weigh chemicals in a laboratory:

1. Standard laboratory balance

2. Analytical balance

Standard Laboratory Balance

Generally, a standard laboratory balance is used for weighing a chemical when an error of ±0.02 g is acceptable. It is also used for pre-weighing, where approximate mass needs to be determined. Precise mass of the chemical is then determined using an analytical balance. Following are the steps in using a weighing balance in a laboratory.

Introduction

To accomplish experimentations in laboratories, sometimes we need microscope to test/validate our ideas/findings. A microscope is needed to examine objects which are very small which cannot be visualised by naked eyes. However, microscopic study requires some expertise in operating the microscope, particularly the sophisticated microscopes, as well as to prepare (peeling, sections, cutting, fixation, staining, dehydration, mounting, etc.) the sample, so that it can be appropriately examined for the anticipated details.

According to the scientific requirements different types of microscopes are used in the laboratories for research purpose. In this chapter, our aim is to acquaint you with the basics of using and handling of microscopes, particularly a compound microscope that is commonly used by the graduate and post graduate students for basic research.

Microscopy is often designated as the scientific investigation that is employed to review and analyse minute structures and objects using a microscope. The term “microscope” has been derived from ancient Greek words “micros” meaning “small” and “skopeîn” meaning “to look at or see,”.

Microscope

A microscope is a laboratory instrument used to examine objects (biological specimens) which are too small to be seen by the naked eye. The microscope must accomplish three tasks:

• Magnification- to provide a magnified image of the specimen.

• Resolution- to separate the details in the image.

• Contrast- to render the details clearly visible to the eyes.

Although we will tnroduce to you the different types of microscopes used for different speciments, our main focus is to discuss about compound microscope, which is commonly used in the laboratories. Compound microscope was developed around the beginning of 17th century.

Since mouth pipetting of liquid reagents in a laboratory is not advisable, different tools are needed for safe, easier, and precise dispensing of liquid reagents. This chapter is devoted to make you aware of the tools like vacupette and micropipette which can be used for the handling of liquid reagents in a safe and precise way.

Vacupette

Glass pipettes have been traditionally used by sucking liquid solvent/solution directly using mouth. For safety reasons, mouth pipetting is no longer permitted in a laboratory. Hence, to fill a pipette vacupette is used by placing it on the top of glass or plastic pipette. It acts as a device to create vacuum for filling reagent in the pipette as well as for releasing the reagent with mechanical regulation on the flow out of the liquid. Vacupette can be of different types.

1. Rubber Bulb Vacupette

Rubber bulb vacupette is often used to transfer (pump up and discharge) liquid using a pipette when a larger quantity of chemicals is to be transferred. Small/tiny rubber bulb is attached to glass/plastic pipette for drawing little amount of reagents. Small vacupette is used to precisely discharge smaller volume/ drops.

Handling of Vacupette

• Insert the top of a pipette in the bottom of the rubber bulb vacupette (Figure 6.1).

• Remove air from the pipette by squeezing valve “A” at the top of the vacupette while simultaneously squeezing the rubber bulb. The amount of air released depends on the size of the rubber bulb.

Different basic as well as advanced laboratory equipment need to be used in a laboratory while conducting experiments. Many of them like pH meter, laminar air-flow hood, viscometer, thermometer, magnetic stirrer, microwave ovens, incubators, sand-bath, water-bath, oil-bath, etc. are the basic and simple equipment which require to be used routinely while conducting experiments in a laboratory. However, basic knowledge of their operation, handling procedure, when to be used, how to be used, and more importantly what not to be done with it, etc. are essential for keeping them in serviceable condition as well as for the accuracy/precision in their operation. In this chapter, we will discuss about some of the routinely used laboratory equipment along, while some other sophisticated equipment will be discussed in other chapters.

pH Meter

A pH meter measures exactly the H+ concentration of a solution; thus indicates acidity or alkalinity of a solution with greater precision. Actually, it measures the difference in electrical potential between a pH electrode and a reference electrode, and display the results converted into the corresponding pH value. Electrode is the key part (rod-like structure, usually made of glass) of a pH meter, having a bulb at the bottom containing a sensor. The pH meters range from simple and inexpensive pen-like devices (portable/pocket pH meter) to complex and expensive laboratory instruments with computer interface (Figure 5.1).

Frequent calibration of a pH meter using the solutions of known pH (standard solution of pH 4.01, 7.00, 10.01, Figure 5.2), generally before each use, ensures the accuracy of the instrument. For very precise work the pH meter should be calibrated before each measurement. For normal use calibration should be performed at the beginning of each day. This is necessary because the glass electrode does not give a reproducible measurement over a longer period of time.

A variety of glassware like burettes, pipettes, measuring cylinders, flasks, separating funnel etc. are used for experimental purposes in a laboratory. All of these need to be used and handled correctly, otherwise the chances of experimental error and damage to the glassware increase significantly. Being made of glass, they need special care while washing and using in experiments because even a minor mistake in handling of glassware may result in breakage of the apparatus. With this precautionary note, let us discuss about some of the specific glassware used in the laboratories.

Burette

Burette is a long graduated glass tube with a stopcock at one end for delivering a known/desired volume of a liquid, especially in titration experiments. It contains a stopcock at the lower end and a tapered capillary tube as the stopcock’s outlet. The flow rate of liquid from the tub stopcock’s outlet can be controlled with the help of stopcock valve. Burette is used for measuring the volume of a solution/chemical added in a quantitative (titrimetric) estimation experiment.

Procedure of using a Burette

The burette reading must be noted before and after delivering the chemical/ solution from the burette. The difference in these two readings is the volume of the chemical/solution released out of the burette. The solution should be released slowly/drop-by-drop to avoid any experimental error/mistake. If the solution/chemical is allowed to pour out too fast, the wall of the burette will not drain properly and some liquid may remain sticking to the surface of the walls. Moreover, excessive amount of chemical may be released/dropped during the experiment, particularly during the titration experiment wherein we need to know the exact amount of acid/base needed to get neutralization point. Thus, any minor mistake may lead to get a faulty reading. The measuring capacity of a burette usually used in a laboratory is 50 mL.

Introduction

A variety of glassware, plasticware, and other apparatus are used for experimental purposes in a laboratory. They need to be washed, cleaned, dried, and sometimes sterilized before they can be used in the next set of experiments. Glassware needs special care of handling during washing, drying, sterilization, etc. because even a minor mistake in handling of glassware may result in its cracking/breakage and it may become unserviceable. All of these may result in a delay in experimentation if spare piece of the glassware is not available or it may affect serviceability of the associated apparatus.

A large number of glassware such as beaker, flask, measuring cylinder, burette, test tube, etc. are regularly used in a laboratory, which requires careful handling during the different stages of experiment as well as after completion of the experiment while preparing them for another set of experimentation.

Glass being a complex silicate, its properties depends on the type of silicate anions in the structure and the cations formed. Thermal properties of glass changes by the addition of boron oxide (B2 O3 ); hence, borosilicate glass is extensively used in the laboratories. With this brief background information, let us discuss about the necessity, procedures, and precautions during washing, cleaning, drying and sterilization of laboratory-wares. Figure 3.1 shows various advantages of using borosilicate glass in a laboratory.

Good laboratory practices require clean glassware, because an experiment may give erroneous result if dirty glassware is used at any step of the experiment. Hence, the glassware must be physically and chemically clean; and in many cases, it must be microbiologically clean or sterile. The commonly used glassware in a laboratory include test tube, measuring cylinder, pipettes, beakers, flasks, microscopic slides, etc.

Introduction

While experimenting with chemicals, sometimes we need to dry the chemical/ solution (solid, liquid or gas) using a process like evaporation or distillation, mainly to remove water or different solvents. Drying is the process of evaporation or distillation to remove water/solvent from a solution, solid or even from gas. Generally, a drying agent (usually an anhydrous inorganic salt) is used for this process, which reacts with the water (present in the chemical/ solvent to be dried) to form a hydrate. For example, anhydrous MgSO4 is one of the drying agents. The drying agents are recognized by their capacity (the rate at which they absorb water) and their intensity (the amount of water left behind in the solvent at equilibrium). No chemical reaction or change in phase occurs during the drying process.

Type of Solvents

Apart from water (a universal solvent), there are three families of organic solvents used in laboratories.

1) Oxygenated solvents: Alcohols, Ketones, Glycol ethers, Glycol esters, etc.

2) Hydrocarbon solvents: Aliphatic and Aromatic hydrocarbons.

3) Halogenated solvents: Chlorinated hydrocarbons.

Introduction

For proper and efficient experiment to be conducted, experimenters need so many chemicals to deal with. This includes different acids, base, salt, different kind of indicators and so on. Some of these chemicals are safe for health, but some of them are dangerous to health when come in close proximity to our body while some of the chemicals cause health issues on inhaling. These may cause improper functioning of some of our physiological activities. Acids and bases are very dangerous for our tissues when come in our contact. These may cause serious respiratory problem in addition to injuries on coming to our contact. Some chemicals are exploratory yet they may cause even death of the person working with them in laboratory. Therefore, mere use of the chemicals in laboratory is not only the objective of the experimenter. The person working in laboratory should also be concerned about the proper use and handing of the chemical while working with it. Thus, every researcher of an institution who has to work with chemicals in a laboratory should be made aware about the chemicals to be used in the laboratory and the consequences if any mishandling of a laboratory chemical occurs. Not only these, suitable precautionary measures should also be known to the researcher along with ensuring the availability of the required safety equipment/installations. Hence, a proper understanding of the ways of handling of chemicals in a laboratory is of prime importance for the personals like you who are working or have to work in laboratory.

Safe Handling of Chemicals in Laboratory

Safe use/handling of chemicals in laboratory include minimized exposure to chemicals, maintaining safety data sheets, proper labelling, storage, segregation, and transport of chemicals for personal hygiene.

As the present study draws to a close, it opens up a myriad of possibilities for future research and exploration in the field of the present study. The insights gained and the findings presented in this thesis may provide a foundation for building upon and extending the existing body of knowledge. By identifying key areas that require additional attention and offering suggestions for future studies, this section of the chapter aims to inspire and guide researchers in their pursuit of enhancing our understanding and driving progress in this dynamic and evolving field.

The present study leaves behind the following domains which may be researched out in future:

1) The present research can be replicated with varied geo-spatial situations viz., in different locations and settings, with different respondents, with spatio-temporal expansion and methodological innovation.

Despite the meticulous methodology and extensive analysis employed throughout this research project, it is crucial to acknowledge and address the limitations that arose during the study. By recognizing these limitations, we not only ensure a transparent evaluation of the research process but also shed light on potential areas for future investigations to refine and expand upon the findings.

In this section of the chapter, the limitations encountered in this thesis are highlighted as follows:

7.1 Summary

Climate change is a complex global phenomenon that poses significant challenges for agriculture including uncertainty about future weather patterns, soil conditions, and other environmental factors. Farmers are facing the difficult task of interpreting and responding to these changes in order to maintain their livelihoods and ensure food security for growing populations. In this context, mental modelling has emerged as a useful approach for understanding how farmers interpret and respond to climate change. Mental models are cognitive representations of how individuals understand and make decisions about complex systems, and can be used to identify key drivers of behaviour and decision-making.

The present study has explored the social, ecological, and economic factors that influence farmers’ mental models on perception of climate change, level of uncertainty and trend of agricultural change in the selected study area. It argues that farmers’ mental models are shaped by a range of factors, including their own experiences and observations, social networks and interactions, access to information and resources, and broader economic and political contexts. The present study has also highlighted the role of uncertainty in shaping farmers’ mental models and decision-making. Uncertainty about the future impacts of climate change can create significant challenges for farmers, as they must make decisions based on incomplete information and uncertain predictions.

The current research work was carried out in the new alluvial zone which is one of the agro-ecological regions in the state of West Bengal. To conduct the study, three villages located in the Haringhata block of Nadia district were chosen randomly. A total of 90 farmers were selected with 30 respondents each from the villages of Kastadanga, JalkarBhomra and Subuddipur. Initially a list of contact farmers was collected from the Kastadanga SKUS and few contact farmers were selected randomly from the list. Afterwards Snowball sampling method was followed to select respondents.

This chapter focuses on presenting empirical findings from data analysis to provide sufficient interpretation so to draw reliable conclusions. To enhance and complement the textual contents of the study, tables and models are frequently utilized. Additionally, the discussion serves to contextualize the study’s findings and establish connections with the research work.

The pattern of distribution of the variable, Education (x2 ) depicts that the maximum education of the respondents is 5 and the minimum is 2. The mean value of education of the respondents is 4.09 with standard deviation of 0.82. The Coefficient of Variation is 19.96 per cent which indicates the distribution of variable is highly consistent.

the maximum functional education of the respondents is 15 and the minimum is 2. The mean value of functional education of the respondents is 8.27 with standard deviation of 3.39. The distribution of the variable is fairly consistent with a CV value of 41.06 per cent.

The pattern of distribution of the variable, Size of family (x4 ) depicts that the maximum size of family of the respondents is 9 and the minimum is 3. The mean value of size of family of the respondents is 5.62 with standard deviation of 1.48. The Coefficient of Variation is 26.34 per cent which indicates the distribution of variable is fairly consistent.

The pattern of distribution of the variable, Cultivated Land Holding per capita (x5 ) depicts that the maximum cultivated land holding of the respondents is 1.92 and the minimum is 0.25. The mean value of cultivated land holding of the respondents is 0.77 with standard deviation of 0.30. The distribution of the variable is fairly consistent with a CV value of 38.83 per cent.

the maximum no. of land fragment of the respondents is 2 and the minimum is 1. The mean value of no. of land fragment of the respondents is 1.24 with standard deviation of 0.43. The Coefficient of Variation is 34.73 per cent which indicates the distribution of variable is fairly consistent.

The pattern of distribution of the variable, Avg. size of land fragment (x7 ) depicts that the maximum avg. size of land fragment of the respondents is 1.25 and the minimum is 0.25. The mean value of avg. size of land fragment of the respondents is 0.64 with standard deviation of 0.19. The distribution of the variable is fairly consistent with a CV value of 30.18 per cent.

The research methodology constitutes a comprehensive and well-structured plan of action for conducting the investigation and serves as a blueprint for the research process. Its purpose is to gain an understanding of the underlying concepts, methods, and techniques employed to define the study, gather relevant information, analyse data, and interpret the findings, thereby revealing concrete evidence and formulating theories.

In this chapter, an in-depth exploration of the research methodology is undertaken 
to comprehend the concepts, methods, and techniques employed for designing the 
study, gathering data, conducting data analysis, and interpreting the results. To 
facilitate clarity and ease of comprehension, the chapter has been divided into the 
following sub-headings:
 1. Research Design
 2. Locale of research
 3. Pilot study
 4. Sampling Design
 5. Variables and their Empirical measurement
 6. Methods of Data collection
 7. Preparation of Interview Schedule
 8. Item analysis and Expert’s rating for devising scoring technique for variables
 9. Pre-testing of Interview Schedule
 10. Data Processing and Analysis for FCM
 11. Statistical Tools used for Data Analysis
 12. Software used for Data Analysis 5.1. Research Design
A research design provides a structured approach for researchers to answer their research questions in an unbiased, precise, and cost-effective way. It establishes 
a framework for testing relationships between variables and ensures that the 
research is going on right direction. There are various types of research designs available, and the current study uses the descriptive and diagnostic research design. 
Descriptive research design is used to collect accurate and relevant information 
about the current state of a phenomenon, and to draw valid and generalized 
conclusions wherever possible. 

The research setting provides specific details about the location where a study is conducted. This study area typically encompasses a particular geographical region, such as a state, district, block, or gram panchayat, chosen according to the research requirements. The researcher must have a thorough understanding of the locality, including its communication methods, in order to effectively access all parts of the region for data collection.

In social science research, a clear comprehension of the characteristics, attitudes, and behaviours of the individuals within the study area is essential. Without this understanding, accurately conceptualizing, perceiving, and interpreting data becomes challenging. To obtain a comprehensive understanding of the implications and behavioural intricacies of the local people representing a broader community in the referenced study area, researchers must have a solid grasp of these factors. Therefore, this chapter critically analyses the socio-demographic background of the residents in a rural setting.

The term “research setting” denotes the context in which research inputs and elements are absorbed, interacted with, and mutually contribute to the system’s overall performance.

The research setting is crucial because it shapes and influences the interactions of various factors and components. Therefore, conducting a study on how farmers perceive persuasive issues and other topics requires a local context that takes into account the unique natural environment, demographics, crop ecology, institutional framework, and socio-cultural factors. For example, when writing a paper about the social behaviour of chimpanzees, it is important to include details about the research setting. This may include information such as where the chimpanzees were studied (in the wild or in captivity), the number of chimpanzees that were observed, whether they were part of the same social group, the geographic location, the time of the study, the season/weather conditions, the availability of resources such as food, water, and shelter, and any environmental factors that were present. There are two types of research settings: macro and micro. The macro research setting encompasses the entire state, while the micro research setting focuses on specific districts, blocks, or village peripheries. The aim of this approach is to understand the study environment from broader perspectives using the state as a reference frame and the district and block profiles as units of in-depth analysis.

The Review of literature is a systematic, explicit and reproducible method of identifying, evaluating and synthesizing the existing body of a completed and a recorded work produced by researchers, scholars and practitioners. The Review of literature is also a resource paper which includes the current knowledge including substantive findings as well as the theoretical and the methodological contributions to a particular topic. These are available mostly in academic and scholastic journals.

Review of literature provide the basics for research in nearly every academic f ield. This chapter is an extensive survey of all past studies relevant to the field of investigation which gives knowledge of what others have found out in the related field of study. In any scientific investigation or research, a comprehensive presentation of the review of literature is a statutory requirement.

The aim of review of literature is to highlight what has been done so far in the field of interest and how our findings are related to earlier research.

Its main functions are to develop a better understanding of the problem to be investigated, delineate a new area of study and avoid unnecessary duplication, decide on the tools and techniques to be adopted including developing some new ones and also to relate the present study with the previous ones by finding out the areas of agreement and disagreement.

The review of literature may

(a) Indicate the following by providing referential inputs – Approaches, Methods, Variables used, Statistical procedure, Expected results.

(b) Provide theoretical background to the study.

(c) Help to justify how the findings are related to the body of knowledge in the f ield of research.

(d) Establish a link between what we have proposed to study and what has already been found. It helps us to refine the research methodology.

In the context of a thesis, theoretical orientation refers to the conceptual or theoretical framework or perspective that informs and shapes the overall approach and analysis of the research (Natalie, 2022). It involves selecting and adopting a specific set of theories, concepts, or models that provide a foundation for understanding and interpreting the research topic. The theoretical orientation helps to guide the methodology, and provide a coherent framework for organizing and analysing the data.

Theoretical orientation in a thesis serves several purposes. It helps to establish a theoretical basis for the study, providing a context for the research and connecting it to existing knowledge and theories in the field. It also helps to guide the selection of appropriate research methods and data analysis techniques. The theoretical orientation shapes the overall structure and content of the thesis, ensuring that the study is grounded in relevant theories and concepts and contributes to the existing body of knowledge. By adopting a specific theoretical orientation, the researcher can provide a clear and systematic framework for understanding and interpreting the findings of the study. This orientation also helps to establish credibility and validity for the research, as it demonstrates the researcher’s awareness of and engagement with the existing theoretical perspectives in the field.

This chapter addresses crucial terminology and concepts essential for effectively communicating the ideas of the current study. It lays the groundwork for empirical investigation and aids in the selection of predictor variables. Additionally, this section involves a scholarly and theoretical exercise that influences the operational definition and content of the research. It combines conceptual inputs, axiomatic inputs, logical discussions, and the pursuit of knowledge to construct a theory that aligns with the research objectives and designs.

Climate change is a complex and dynamic phenomenon that has significant impacts on agriculture, especially in terms of uncertainty and variability. Agricultural dynamics refer to the various factors that affect agricultural production, including climate, soil, water availability, pests, and diseases, as well as market forces and government policies. Climate change has significant impacts on agricultural dynamics. Variations in temperature, patterns of rainfall, and occurrences of extreme weather can influence crop yields and livestock production in both beneficial and detrimental ways (UCAR, 2021). The effects of climate change on farming can differ based on the geographical location and the specific agricultural practices employed (Wikipedia Contributors, 2019). For example, in some areas, increased rainfall may lead to increased crop yields, while in other areas, droughts may lead to crop failures and loss of income for farmers. Additionally, rising temperatures can cause reduced crop yields and decreased quality of some crops.

Uncertainty is a term used to describe a situation where there is a lack of information, knowledge, or predictability about a particular outcome or event. In agricultural dynamics, uncertainty can refer to a range of factors that can affect the production, distribution, and consumption of agricultural products. One of the most significant sources of uncertainty in agriculture is the weather. The productivity of crops can be significantly influenced by weather conditions, including rainfall and temperature and farmers need to effectively adjust their agricultural practices to adapt to these changing weather patterns (Guntukulaand Goyari, 2020). For example, droughts or floods can reduce crop yields, while frost or hail can damage crops(Lamaoui et al., 2018).

The social, ecological and economic interpretation of climate change by farmers involves understanding the interactions between the natural environment, social systems, and economic factors (Spash, Clive L, 2011). Farmers need to consider these interactions when making decisions about their farming practices. For example, changes in climate can affect the availability of water, soil fertility, and crop yields, which can in turn affect social and economic factors such as food security and income. Farmers need to take into account the wider consequences of their agricultural practices on society and the environment which includes considering the impacts on biodiversity, ecosystems, and human well-being (Warsaw et al., 2021).

Spices are those plants, the products of which imparts strong flavour and aroma to the food. This is attributed to the presence of compounds such as volatile oils present in the plant parts. India is renowned for wide variety of spices and is designated as land of spices. Spices are indispensable ingredient in almost all Indian dishes and cuisines foreign invaders invaded India for spices in ancient times. India is figured as the largest producer, consumer and exporter of spices and spice products in the world. Turmeric and Ginger are the major rhizomatous spices of vegetable origin widely cultivating and using in India. They are known for its antimicrobial and antioxidant properties, hence using in medicines for the treatment of various ailments. Apart from these rhizomatous spices, India is also contributing a major share in the production of seed spices. Around twenty spices among sixty-three spices growing in our country are classified as seed spices. Major seed spices of India are coriander, cumin, fennel, fenugreek, dill, ajowain, celery, anise, nigella, caraway etc. These seed spices are extensively cultivated in the arid to semi-arid region of India as Rabi crop.

It is estimated that, spice production in our country increased from 67.64 lakh tonnes in 2014-15 to 106.79 lakh tonnes in 2020-21 with an annual growth rate 7.9%. There is a corresponding increase in the area of cultivation also. An increase in area from 32.24 lakh hectare to 45.28 lakh hectare was observed. Among the major spices, cumin (14.8%), garlic (14.7%), ginger (7.5%), fennel (6.8%), coriander (6.2%), fenugreek (5.8%), red chilli (4.21%) and turmeric (1.3 %), showed significant growth rate in production. The export of spices contributes around 41% of the total export earnings from all horticulture crops in the country and it ranks fourth among agricultural commodities, after marine products, non-basmati rice and basmati rice.

Exotic vegetables are those vegetables that are introduced or not common in India. Its area under cultivation is comparatively less and most of the growing area surrounds the targeted markets especially five star hotels and restaurants. Now a days, there is an increasing trend in cultivation of these high value vegetables because of increased demand and good profit. Farmers could sell these vegetables at a at a higher price when compared to traditional vegetables. They are mostly cultivated in net houses or poly houses. Some of the exotic vegetables popular in India are asparagus, celery, broccoli, lettuce, parsley etc.

Asparagus (Asparagus officinalis)

Asparagus is a dioecious monocot which produces tender succulent above ground stems known as spears from fleshy underground crown. These tender spears are used as vegetable, for cooking purpose and for the preparation of soups. The white crystalline substance, asparagine present in the juice of young shoots have diuretic properties and is used in cardiac dropsy and chronic gut. Asparagus is mainly cultivating in countries like USA, Germany, Spain, France etc. while in India, it is cultivating only in a negligible area. Asparagus is grouped under cool season vegetables. But newly emerging spears are susceptible to frost though a wide range of environmental conditions can be tolerated by mature asparagus. Asparagus is native to temperate Europe and Asia and has been cultivating for 200 years.

Morphology

Asparagus is perennial in nature and produce erect branched stem and grows upto a height of 1 to 3 m. The crop yield continuously for 10-15 years from the initial year of spear production. Asparagus roots are tuberous and the leaves are narrow and needle like. Male and female flowers are produced in different plants.

Leafy vegetables are rich source of vitamins like A, C and minerals like calcium and iron. Most of the leafy vegetables are quick growing types and shows good response to manures and fertilizers. Their easiness in cultivation and adaptation to diverse agroclimatic conditions makes it suitable for various cropping systems. Some of them are annual in nature while some others are perennial. Leaves as well as tender stems are uses as vegetable in the cooked form. They are reported to have antinutritional factors, oxalates and nitrates compared to other vegetables. The annual leafy vegetables are widely cultivating in our country and among them, amaranth, palak, spinach and fenugreek are the major ones. These crops are short duration in nature, completes their entire life cycle by 80-90 days and produce good yield and brings monetary returns to farmers in short time.

Amaranth (Amaranth sp.)

It is herbaceous fast growing annual leafy vegetable grown for leaves as well as grains. Vegetable amaranth is reported to be equal or superior in taste to that of spinach. Leaves and succulent stem are good source of vitamin A, vitamin C, calcium and iron and are used as vegetable. High oxalate and nitrate content are reported in some species of amaranth.

The major vegetable species of amaranth cultivating in India is A. ticolor. It is suggested to be originated in South or South east Asia particularly in India. Another vegetable type, A. dubius shows diversity in Central America, Indonesia, Asia and Africa. A. lividus is the popular vegetable type in South and Central Europe. Other leafy species are A. viridis, A. tristis and A. spinosus. A. cruentus, A. hypochondriacus and A. caudatus are cultivating for grain purpose. Taxonomists suggested two sections in Amaranthus namely blitopsis which include leafy types and amaranthus which constitutes grain types.

Potato, the third most important food crop in the world after rice and wheat is becoming the main source of food and income in developing countries. India stands second in production and area of potato in world after China leaving behind the Russian Federation. In India, potato is an important component in cereal and vegetable-based cropping systems for its ability to produce high dry matter/ha in short duration and high yield compared to cereals. It is grown as mixed crop, intercrop, relay or sequential crop along with other vegetables, cereals and cash crops.

Potato is originated in South America in the high Andean hills and towards the end of 16th century it was first introduced into Europe by Spanish conquerors. In the early 17th century, it was brought to India by British missionaries or Portuguese traders. Thus, potato developed as a temperate crop and was later distributed throughout the world. Cultivated potato consists of two sub species, Solanum tuberosm ssp. andigena and S. tuberosum ssp tuberosum. Subspecies andigena is the primitive form which is adapted to shorter day length and high altitudes of mountains (above 2000). The presently cultivating tuberosum subspecies is adapted to long or short day condition of tropical regions and lower altitudes (500-200m above MSL) and is believed to be evolved from andigena types through selection.

Morphology and Floral Biology

Potato is herbaceous in nature with erect to prostrate stem. Stem is variably coloured from green, purple or mottled green or purple. Leaves are pinnate which ends in single terminal leaflet. Stolon arise from the portion of the plant below the ground. The stolon bears rudimentary leaves and tip is hook shaped. The sub apical portion at the tip of stolon enlarges to produce tubers. Hence, potato tubers are the modification of underground stem with shortened axis and poorly developed eyes. The leaf scar with a subtended lateral bud having underdeveloped internodes is referred to as the eye of the tuber. Potato is a long day plant. Moderate humidity and long day condition favours flowering. Hence flowering and fruit set is observed in hilly tracts. Inflorescence is produced terminally and is usually cymose. Flowers are bisexual, actinomorphic, and hypogynous with 5 calyx, 5 corolla and 5 stamens. The colour of corolla varies from white, pink, lilac, blue and purple. Pollen shedding is through the pores at the tip of the anthers. Mature fruit is a berry. Self-incompatibility is observed in diploid species and tetraploids are self-compatible.

Root crops are cool season vegetables growing for its fleshy root developed from the tap root and hypocotyl region. They are grown during winter in tropical and subtropical condition, throughout the year in hills and spring-summer in high altitude hills. The major root crops growing in our country include carrot (Umbelliferae), radish (Brassicaceae), beet root (Chenopodiaceae) and turnip (Brassicaceae). Two distinct types viz., asiatic or tropical types and European types present in these root crops except beet root. In beet root, European or biennial types only exist. The tropical types are annuals that does not require chilling treatment for flowering. Therefore, seed production of these types is taken up in plains but European types are biennials and they require vernalization treatment after vegetative phase for flowering. Hence, the seed production of biennial types is possible in hills. The seed productions of Asiatic varieties of root crops are done in Punjab, Haryana, U.P. and parts of Rajasthan and Gujarat. Whereas, the seed production of temperate varieties is done in Saproon valley, Kullu valley, Kalpa valley of H.P., Kashmir valley of J&K and selected location of Northeast region specially Sikkim and Arunachal Pradesh. The seed production of beet root is being done in high altitude hills of J&K. All the crops belonging to this group are similar in the cultural practices though they come under different family. All root crops prefer well drained loose and friable soil. Ridge or bed planting is mainly following in all root crops. They are direct sown crops and does not respond to transplanting. They are highly cross pollinated.

The two methods of seed production following in root crops are seed-to-seed method and root-to-seed method. The first method is followed for the production of truthfully labelled seed or certified seed as the seed yield will be more, but quality remains inferior to the second method. The breeder seed as well as foundation seeds require utmost quality hence root-to-seed is preferred for its production. In this method, the matured roots are harvested and the roots are inspected for its trueness. Only true-to-type roots are selected for replanting in the field for seed production and undesirable ones are discarded. The selection and rouging in seed crop are done on the basis of foliage characters and external and internal root characters (colour, shape, size, flesh colour, core size, pungency, indistinct rings etc.). The roots after selection are prepared for replanting by cutting one-third top (shoot) portion and one-fourth to one-half root cut. the prepared stecklings are replanted to obtain good yield of quality seeds. In case of turnip and beet, after selection, the tap root and tops of the roots are trimmed without injuring the crown portion and is replanted.

Bulb Crops

Bulb crops are cool season vegetables that are grown during the winter months in plains. They are mainly growing for its fleshy leaf bases or fleshy axillary buds or blanched stem and leaves. Major bulb crops that are widely cultivating in our country comprises of onion, garlic and leek. They possess a characteristic odour due to the presence of volatile oil, allyl propyl disulphide (onion) and diallyl disulphide (garlic). They are either seed propagated as in onion and leek or through cloves as in garlic or through bulblets in multiplier onion. Seed propagated bulb crops are annual for bulb production and biennial for seed production. Seed production of bulb crops can be undertaken in plains especially in the central part of the country and southern plains whereas, northern plains hold the problems of Stemphylium and purple blotch during the seed crop and subsequent lower seed yield makes northern plains unfit for seed production. But these areas can be utilized for seed production by following the delayed planting of seed crop and avoiding the climate favouring the diseases.

Onion (Allium cepa)

Onion is a popular vegetable grown for its pungent bulbs. Bulbs are consumed as raw vegetable, as cooked vegetable or in preserved forms. Onion is reported to be originated in Central Asia and Mediterranean region is considered as the secondary centre of origin. It is adapted to a wide range of climate and it is being cultivated in India in three seasons viz., kharif, late kharif and rabi. India is having the largest area under onion cultivation in the world. It is also contributing substantially towards the export of the value-added products from our country. White or yellow onions are commonly using for producing dehydrated products whereas, the red ones are more preferred for direct consumption.

Cole crops refers to vegetable crops originated from a common ancestor, colewort or wild cliff cabbage (B. oleraceae var oleraceae or Brassica sylvestris L.) and they belong to family Brassicaceae. They prefer low temperature for growth and development hence mainly grown as temperate vegetables in temperate and subtropical climatic zones. This group of vegetable includes cabbage, cauliflower, knol khol, brussels sprout, sprouting broccoli, kale, Chinese cabbage etc. They are Mediterranean vegetables since its centre of origin is considered to be Mediterranean region. Later on, it spread into different parts of the temperate and subtropical countries. In India, cultivation of cole crops was started in the hills where temperate climate prevails and in states like Uttar Pradesh, Orissa, Bihar, Assam, West Bengal, Maharashtra and Karnataka as cool season crop. But with the introduction of tropical types which are more tolerant to high temperature cultivation, cole crops are taking new pace in the cropping system of nontraditional areas. Cabbage and cauliflower became the most widely grown cole crops though cabbage, knol khol and curly kale were the first cultivated types and cauliflower came in the mid-way followed by brussels sprout and sprouting broccoli. The genetic designation of different vegetables under cole crops were described by several workers of which Linnaeus (1753) designated all the cole crops under one species, Brassica oleraceae and in 1969, Neuwhof classified them into different forms as B oleracea var capitata L. (cabbage); B oleracea var botrytis L. (cauliflower); B oleracea var gemmifera DC. (Brussels sprout); B oleracea var italica L. (sprouting broccoli); B oleracea var gongyloides L. (knol khol) and B oleracea var acephala DC. (kale).

Cole crops are usually biennial herbs with tap root system and erect stem. They are highly cross-pollinated and temperature sensitive for the production of head and curd. They are classified as early, mid and late varieties based on the temperature requirement and duration. Among this, early varieties are found to be high temperature tolerant and late varieties are low temperature tolerant. The flowering in cole crops requires a chilling temperature of 4-70C for about 6 8 weeks which transforms the juvenile phase into reproductive stage with exceptions of some early varieties of cauliflower and sprouting broccoli. Upon receiving the chilling temperature, the central stalk will develop into reproductive shoot or multiple reproductive shoots will be formed from the curd. The earlier is seen in cabbage, brussels sprout and kale whereas, the latter in cauliflower and sprouting broccoli. The seed production of cole crops belonging to temperate groups are mostly undertaken in temperate hills especially in the hills of Uttaranchal and Himachal Pradesh whereas, seeds of tropical types are produced in plains and lower hills of India.

Leguminous vegetables include wide range of vegetables which can grow in climatic conditions extending from warm to cool weather. They are rich in protein, essential amino acids and fibre hence, recognized as an important component in ensuring nutritional security. They are growing for its pods for vegetable purpose, seeds as pulse or for dual purpose. They are excellent green manures that fix atmospheric nitrogen into soil and cultivation of leguminous vegetable crops is recommended as an integral component in sustainable soil fertility management. Leguminous vegetables include tropical types namely cowpea, hyacinth bean, winged bean, jack bean, sword bean while, cool season legumes include French bean, garden peas, lima bean and broad bean. They are mostly self-pollinated owing to its cleistogamous flowers. Pole or bush types are available in these crops and all are seed propagated. Among the different leguminous crops, French bean, garden peas and cowpea are under commercial cultivation while other leguminous vegetables are region specific in cultivation.

Cowpea (Vigna unguiculata)

Cowpea is a warm season legume grown both as vegetable crop as well as pulse crop. Tender pods are used as vegetable and dry beans are used as pulse. Cowpea is mainly growing for its seeds as pulse than as vegetable crop. The vegetable types bear stringless pods of more than 30 cm long, fleshy pericarp and higher monosaccharide to polysaccharide ratio. Cowpea is believed to be originated and domesticated in West or Central Africa especially Nigeria where diversity was noticed abundant. The cultivated cowpea, Vigna unguiculata is divided into different cultivar groups viz., pulse type as Vigna unguiculata var. unguiculata, Vigna unguiculata var. cylindrica or var. biflora as dual purpose type for fodder and dry seeds and vegetable type as Vigna unguiculata var. sesquipedalis (yardlong bean/string bean/asparagus bean). The cultivar groups, biflora and sesquipedalis are suggested to be originated from ssp. unguiculata in India and South East Asia respectively.

Okra is a tropical vegetable cultivating for its tender fruits. It is one of the major vegetable crops in India which can be grown in summer as well as rainy season. Fruits are rich in vitamin A, vitamin C and minerals like calcium, phosphorus, iodine, iron, and potassium. Dry fruit skins and fibers are valued for manufacture of paper and card boards. The processed product, dehydrated okra plays a considerable role in export earnings. Dried seeds of okra found to have 20-24 percent protein and 13-22% edible oil hence, used for edible oil production. Mucilaginous stem is used for clarification of cane juice for preparing jaggery. Roasted ground seed are used as coffee substitute.

Cultivated okra is believed to be originated in tropical and subtropical Africa and phytogeographic studies indicated that it arose in Abyssinian region (Ethiopia). However, existence of large number of related species with wide variability and dominant characters suggested India as the secondary centre of origin. Some researchers suggested Hindustani centre of origin mainly India, Pakistan, & Burma.

Morphology and Pollination

Okra is an erect robust herbaceous annual with green or green with reddish tinged stem. Leaves are alternate, 3-7 lobbed, palmate, hirsute and serrate. Flowers are solitary, bisexual, axillary, peduncle is 2 cm long, epicalyx is up to 10 and bracteoles narrow and hairy. Calyx is fused with corolla hence falls with corolla after anthesis. Petals are 5, separate, yellow coloured with crimson spot on claw. Staminal column consists of numerous stamens (80-120) which are united to the base of the petals. Ovary is superior, stigma is small, 5-8 fid and deep red. Fruit is pyramidal and oblong beaked capsule, longitudinally furrowed, hirsute or glabrous, dehisce longitudinally when ripe. Seeds are dark green to dark brown, round and tuberculate.

Cucurbits account for the largest group of tropical vegetables, consisting of cucumber, muskmelon, bitter gourd, bottle gourd, ridge gourd, sponge gourd, snake gourd, pointed gourd, bitter gourd, ivy gourd, wax or ash gourd, chow chow or chayote, pumpkin, summer squash, winter squash, watermelon etc. They all belongs to the family Cucurbitaceae. They are herbaceous in nature and are characterized by tendril-bearing vines and adaptation to warm climate of the tropics and subtropics. Among the different genera of cucurbit vegetables, three genera viz., Cucumis (cucumbers, melons), Cucurbita (pumpkins, squash), and Citrullus (watermelons), are economically important in the world while the others hold regional importance. Most of the cucurbits are grown for its immature fruits and are consumed as salads or cooked vegetable. While, muskmelon and watermelon are used fresh as dessert fruits. Pumpkin and squashes are used in confectionaries besides as cooked vegetables. Mature ash gourd fruits are used in sweet candy preparation.

Cucurbits are low in calories, minerals, or vitamins. However, bitter gourd is rich source of vitamin C, the squash, pointed gourd, and muskmelon contain good amounts of carotene, kakrol is high in protein, and pointed gourd is rich in calcium. Bitterness in some species of cucurbits was attributed by the varying degree of bitter principles, cucurbitacin. Bitter gourd is known for its hypoglycemic activity due to phytoceutical, cheratin in the fruit, which lowers blood sugar level.

Solanaceae is a plant family that comprising of 2300 species and 85 genera of which, nearly one half of the species belongs to the genus Solanum. The genus Solanum includes both tuber bearing and tuber nonbearing forms. Among the tuber nonbearing forms, the major fruit vegetables growing throughout the world are tomato, brinjal and chilli. They prefer warm humid tropical climate for growth and development. Hence, the agroclimatic regions where this type of climate prevails is suitable for seed production of these vegetables. In addition to this, hybrid vigour has been widely commercially exploited in these crops because of easiness in emasculation and pollination and availability of large number of hybrid seeds per fruit.

Tomato (Solanum esculentum L.)

Tomato is the most important warm season vegetable growing throughout the world. The fruit is valued as salad vegetable and for cooking purpose. It is one of the vegetables from which, a wide variety of processed products are made. The crop is grown throughout the year in one part or other part of the country and it has become one of the indispensable fruit vegetables in our diet. It is rich in vitamin A, vitamin C, niacin, and minerals like calcium, phosphorus etc and it is rich in antioxidant, lycopene which is the carotenoid responsible for the red colour of tomato. Tomato is of recent origin, and it is the native of Peru in South America from where it spread to the entire part of the world.

It involves all the steps starting from the extraction till sowing that makes the seed a high quality product for distribution among the farmers for a profitable crop. It reduces the spoilage during storage, increase the shelf life of the seeds, guarantees good germination and field stand thereby reduces the risk of losing the crop at early stages.

Seed Extraction

The seed extraction methods vary with the stage at which the matured fruits are harvested for seed extraction. Some of the vegetable crops are harvested at well ripe stage while some others are harvested at mature dry stage. Proper extraction should be selected taking into consideration the easiness of seed extraction from the fleshy pulp, relatively more seed recovery and less damage to the quality of seeds. There are different methods of seed extraction includes manual method, fermentation method, chemical method and mechanical method.

Manual Method

The manual method of seed extraction varies depending on the harvested fruit type. There are two types of seed extraction in vegetable crops viz., dry extraction and wet extraction.

1. Dry extraction Method

Dry extraction method is following in vegetables where mature dry fruits or mature dried pods are harvested for seed extraction. The harvested dried fruits are kept in sun for 2-3 days. The dried fruits are split open and the seeds are extracted by beating with pliable bamboo stick or by beating against a hard surface. Heavy beating should be avoided to reduce mechanical injury to the seed. Excessive mechanical injury may result in lowering of seed viability. The extracted seeds are dried in the sun between 8.00-11.00 am and 2.00-5.00 pm to reduce the moisture content to a safe level. This is followed in crops like chilli, okra, sponge gourd, ridge gourd, french bean, lablab bean, cow pea, cluster bean etc.

The horticulture sector of our country is following an increased growth in area and production by 9% and 14 % respectively for the past six years (2015-21). It is reported that the growth rate of horticultural crop production accounts for 2.7% per year. Fruits and vegetables are the major sectors contributing to total horticulture production. As per the report of second advance estimates published by Horticulture statistics division, Ministry of Agriculture and Farmers Welfare during 2020-21, our country recorded the highest horticulture production during 2020-21 with an annual production of 329. 86 million tonnes from an area of 27.23 million hectares and the vegetable sector shared around 196.26 million tonnes production from 10.80 million hectare area. India is the largest producer of okra and ginger and second largest producer of potato, onion, cauliflower, brinjal, cabbage etc. Vegetables also contribute to the export market and onion, mixed vegetables, potatoes, tomato and green chilli are the main vegetables exporting from our country. It is estimated an export of worth Rs. 4969.73 crores from vegetables during the year 2020-21.

Importance of Vegetable Seed Production

Though we are leading in many of the vegetables in area and production, our national average in productivity of the major vegetables is far less compared to the world average. The country s estimated vegetable production by the year 2050 is 375 million tonnes from 15 million hectare with an average productivity of 25 t/ha. Low productivity is mainly because the farmers are still relying on old varieties which are less yielding and susceptible to pest and disease. Unavailability of the quality seeds of newly released varieties is one of the main reasons for this production gap. A substantial increase in yield and quality of vegetable crops can be achieved by adopting suitable agronomic practices, plant protection measures and use of quality seed. Now a days, there is an increased demand for good quality seeds of open pollinated varieties as well as hybrid seeds through which an increased vegetable production can be achieved for meeting the requirement of increasing population. So far, very limited number of vegetable hybrids have been developed and released by public sectors and majority of the existing promising hybrids in vegetables are from private sectors.

Technically, seed is the sexually produced matured ovule consisting of an intact embryo, endosperm and or cotyledon with protective covering (seed coat). A true seed consist of an embryonic plant, a store of food which consists of cotyledons and endosperm and a protective seed coat. Seed can also be considered as any material that is used for planting or regeneration purpose. Rather than seed, propagating materials like healthy seedlings, tuber, bulbs, rhizome, roots, cuttings, setts, slips, all types of grafts and vegetatively propagating materials used for production purpose is also referred to as seed. Thus, seeds are of two types namely sexual or botanical seed and asexual seed. Both can be used for raising the crops thus forms the agricultural seeds.

Based on the moisture content present, seeds are of two types, orthodox seeds and recalcitrant seeds. Orthodox seeds are the botanical seeds which contain moisture content less than 20%. Such seeds do not germinate even if the moisture content is more than 20% unless artificially forced to germinate. Recalcitrant seeds contain moisture content more than 20% and such seeds will not germinate if moisture content is less than 20%. These seeds cannot be stored for long and for long storage, special structures or conditions are required. Mostly perennial plant species produce this type of seeds.

Seed is the primary and most critical input in any crop production programme. A quality seed alone contributes around 15-20% towards total production. The efficacy of all other inputs to increase production will be in vain if the quality of the seed is poor. Hence, use of quality seed is crucial as it appreciably increase the productivity of the crop without depending much on other inputs. This adds to the old saying in Manu Smriti good seed in good field yield abundantly .A quality seed can be defined as genetically and physically pure seeds with high germination percentage, free from seed borne pathogens and safe moisture content. This guarantees good germination, vigorous growth, and good establishment of crop in the field.

History of Seed Industry in India

Farmers dependent mostly on traditional indigenous varieties earlier and its seed requirements were met through their own farm saved seeds or through local sources. But the quality of these seed stocks was not assured, and this resulted in drastic reduction in production as well as productivity of the crops. As a result of various crop improvement programmes, improved varieties possessing high yield potential along with additional characters of biotic and abiotic resistance came into the scenario. But the availability of the seeds of these improved varieties among the farmers remained as a problem and it limited the cultivation of these varieties. This led to the development of a well-planned systemized seed production system. Government of India had framed and brought out different legislations for regulating systemized seed production. These laws are framed to make available quality seeds and planting materials to a common farmer and equip him to approach authority for justice. Seed legislations were initiated in the country by establishment of National Seed Corporation (NSC) during 1963 under Ministry of Agriculture and the seed industry was restructured through the nationals seed projects phase I (1977-78), Phase II (1978-79) and Phase III (1990-1991). These initiatives strengthened the seed industry through formulation of seed regulations, establishment of state seed corporations and seed certification agencies thereby assured the supply of quality seeds to farmers

7.1 Introduction

Artificial intelligence (AI) is the imitation of human intellect in machines designed to emulate human activities and cognitive functions. These machines are designed to accomplish tasks that normally require human intellect, such as learning, problem solving, perception, and decision-making (Zhang et al., 2022). Horticultural crops, including fruits, vegetables, nuts, and ornamental plants, are of paramount importance in global agriculture, as they contribute significantly to food security, economic prosperity, and environmental sustainability. Nevertheless, the cultivation of these crops poses numerous challenges for growers, such as fluctuating market demand, insufficient resources, unpredictable climate conditions, and the constant threat of pests and diseases (Nowak & Grant, 2018). In recent years, the agricultural sector has experienced a significant shift owing to advancements in Artificial Intelligence (AI) technologies (Liakos et al., 2018). AI has emerged as a powerful tool for the cultivation of horticultural crops, providing solutions for boosting productivity, sustainability, and efficiency (Shrivastava & Kumar, 2021). By utilizing AI, horticulturists can overcome the traditional challenges associated with crop management, resource allocation, and environmental sustainability, paving the way for a more resilient and productive agricultural sector (Garg et al., 2019). The integration of AI into horticultural crop production encompasses a diverse range of applications, including precision agriculture, crop monitoring and management, predictive analytics, automated harvesting, and genetic improvement (Golhani et al., 2018). These applications leverage machine learning algorithms, remote sensing technologies, and data analytics to analyze vast amounts of agricultural data ranging from soil composition and weather patterns to crop health and market trends (Araus& Cairns, 2014). Through intelligent data-driven insights, AI enables farmers to make informed decisions in real-time, optimize resource utilization, minimize waste, and maximize yields (Kamilaris et al., 2017). This technological convergence offers promising prospects for food production, enabling stakeholders to explore new avenues for growth, resilience, and sustainability in horticultural crop cultivation (Mohanty et al., 2016).

6.1 Introduction

Perennial crops are crucial for maintaining food security, despite making up only 4.2–4.7% of all agricultural acreage (Kreitzman et al., 2020). The value of fruit and nut production reached a value of USD 823 billion globally in 2020, with the top five fruit crops being apples (87 Mt), oranges (78 Mt), grapes (77 Mt), mangoes (55 Mt), and apricots (41 Mt) (Statista, 2020). The tree nut business is growing quickly, although making up a smaller share of the world’s agricultural output. Global tree nut output is expected to reach 4.2 Mt in 2019–2020, with almonds, cashews, walnuts, hazelnuts, and pistachios accounting for 31%, 17%, 21%, 12%, and 14% of the nut market share, respectively (Serna, 2020).

Tree crops require a longer investment period before producing a commercial yield; sweet cherry takes three years to attain a commercial yield, while pistachio takes ten (Rezaei et al., 2019). High-density monocultures of cultivars chosen for their potential yield, fruit quality, and disease resistance are part of optimal tree crop production techniques. These monocultures are maintained under intensive, high-input farming systems (Simon et al., 2017). However, inadequate pollination can seriously impair the quantity and quality of these agricultural products, prompting for meticulous dedication to pollination research and management (Bosch et al., 2021; Forbes et al., 2019).

Effective pollination, and on the contrary, situations in which pollination is restricted or deficient, depend on both internal and external factors (Figure 7.1, Table 7.1). The period of stigma receptivity, the lifespan of the ovule, and the quality and availability of compatible, suitable pollen are all considered as intrinsic factors (Howlett et al., 2015). Agronomic methods pertaining to plant nutrition, orchard layout, pest and disease control, pollinator activity, pollen quality, and weather patterns and their effects on flowering synchrony are examples of extrinsic factors(Toledo-Hernandez et al., 2017). Hatfield and Prueger assert that pollination is one of the phenological stages that is most sensitive to temperature, and as such, it has a significant impact on productivity (Hatfield and Prueger, 2015). Climate has been shown to be a key factor in determining pollination success and production in crops including peaches, apples, and almonds.

5.1 Introduction

The application of chemical pesticides is crucial in horticultural production, but it is also one of the most hazardous agricultural operations. Overapplication can lead to significant environmental concerns. Spraying is vital for reducing harvest losses and improving productivity. According to Cho et al.,(2012), 30-35% of production damages can be prevented by reducing harmful insects and diseases through spraying. Plant protection is essential for ensuring yield quantity and quality. Effective pesticide use is crucial, as excessive application can lead to pests developing immunity, destruction of beneficial insects and natural pest enemies, and increased harmful insect populations. Excessive pesticide use also disrupts pollination, preventing fruit formation. The method of pesticide application is key to targeting the correct quantity and quality, reducing environmental contamination, and increasing sustainability (Abbas et al., 2020). Site-specific application rates in precision horticulture can decrease pesticide use. Only 30-40% of pesticides typically reach their targets, with most lost to the environment, contaminating workers and causing pollution. Protecting plants with chemicals aims to prevent infections and ensure healthy growth.

Spray drift can harm non-target plants and contaminate watercourses, posing a severe and costly problem for horticultural farmers using air-assisted variable sprayers (Seol et al., 2022). Spray drift refers to airborne spray droplets moving beyond the intended application area from aerial or ground-based methods. Studies have found that a significant portion of pesticides is lost to the air and ground, with up to 30-50% of the applied pesticides being lost through various means (Abbas et al., 2020). Modern trends emphasize the importance of sensors and precision agriculture for effective pesticide use. Understanding tree canopy physical features is crucial for effective spray application. This study focuses on recognizing and geometrically classifying canopies for adequate pesticide protection. Variable rate spraying targets only the canopy, enhancing pesticide application efficiency (Rathnayake et al., 2022). Electronic methods and innovative equipment reduce operational and environmental costs. Sensors tracking canopy characteristics and spacing ensure optimal use of inputs, promoting sustainability.

4.1 Introduction

Pests and plant diseases pose serious risks to global agriculture (Strange and Scott, 2005; Oerke, 2006). Plant diseases and pests are the natural disasters that can impede a plant’s normal growth throughout the plant’s whole life cycle, from seed formation to seedling growth and may result in plant death (Liu and Wang, 2021). Diseases reduce yields in a number of important crops by 21–30% globally (Savary et al., 2019), whereas insect infestations reduce crop output by 30–33% annually (Kumar et al., 2019). For the management, accurate disease diagnosis and pest identification are essential. Artificial intelligence (AI) techniques can be a crucial addition to enable the monitoring of plant diseases and pests at a coarse scale, as traditional field scouting of these pests and diseases is labor-intensive, subject to bias, and typically shows low efficiency (Mahlein, 2016).

Agriculture is undergoing a revolution because of artificial intelligence (AI), which is replacing outdated techniques with more effective ones (Talaviya et al., 2020). AI intervention in agriculture is helping farmers increase productivity and reduce adverse environmental effects (Sujawat, 2021). The application of AI techniques has created new opportunities for scientific research as well as new instruments and ways for handling and analyzing vast volumes of data. An important benefit of AI is its capacity to spot patterns in data that are not immediately obvious or available to humans. This allows for previously unattainable new insights and discoveries (Angermueller et al., 2016; Eraslan et al., 2019). AI is currently changing how we work and live, yet its influence on pathology and entomology research is still underappreciated. AI’s cost-effectiveness, precision, quickness of performance, and adaptability are its core concepts in agriculture (Eli-Chukwu, 2019). In agriculture, artificial intelligence not only helps farmers use their abilities, but it also guides them to obtain better quality and larger yields with fewer resources (Khandelwal and Chavhan, 2019). AI-based technology addresses the issues that agricultural sector faces and helps to increase efficiency in all areas. These challenges include crop harvesting, irrigation, soil content sensitivity, crop monitoring, weeding, harvesting, and establishing.