eChapters

1. Biosecurity

Biosecurity is strategic and integrated approach encompasses policy frameworks to analyze and manage risks for food safety, animal life and health and plant life and health including environment.

9.1. Plant quarantine

Legal restriction of movement of plant materials between countries and between states within the country to prevent or limit introduction and spread of pests and diseases in areas where they do not exist.

Plant quarantine regulatory measures are operative through the Destructive Insects & Pests Act, 1914 (Act 2 of 1914) in the country.

ABSTRACT

Young buds of ginger (Zingiber officinale Rosc.) were isolated and cultured on MS media with BA 3 mg l^-1 induced shoot organogenesis. Young buds from aseptically grown cultures were evaluated for the effectiveness on embryogenic callus production on different concentration of auxin in combination with cytokinin. Among the plant growth hormones tested a combination of 2, 4-D 1 mg l^-1 and BA 0.5 mg l^-1 was most effective in inducing and maintaining embryogenic cultures. The somatic embryo germinated on half strength MS medium supplemented with 3 per cent w/v sucrose and 3 mg l^-1 BA.

ABSTRACT

Protoplast isolation in orchids is considered to be a valuable tool for introduction of desirable traits through somatic cell fusion. The present work on the protoplast in Dendrobium var.sonia is undertaken with a view to standardize protocol for isolation and regeneration. Tender leaf mesophyll tissues of one-month-old Dendrobium var.sonia were used as explant for protoplast isolation. The leaf tissues were treated with enzyme mixture of cellulase 1per cent (Onozuka -R from Trichoderma viride) and macerase 0.25per cent (from Aspergillus niger) for 16 hours. Protoplast from the Dendrobium var. sonia was successfully isolated and regeneration of shoots and roots were obtained in MS medium containing BA 5mg l^-1+NAA 1mg l^-1.The plantlets were hardened in a span of five months.

ABSTRACT

Wetlands play an important role in provisioning of numerous goods and services for mankind. However, the type and the quantum of ecological services differs amongst wetlands. In the present study, we documented the various plant resources used by the riparian communities of two well-recognized wetlands, Dhir beel and Diplai beel, located in Brahmaputra Valley of Assam. The findings of study indicate that the riparian communities are dependent on wetlands for various purposes. The riparian communities uses 48 plant species as vegetables, fruits, fodder, fuel wood, medicine, roofing materials, craft making materials, bio-fertilizer and fish poisoning agent. Some of the plant species such as Enhydra fluctuans, Lasia spinosa, Colocasia esculenta, Diplazium esculentum, Ipomoea aquatica and Monocharia hastata aresold in the village market as local vegetables. The wetlands generate income for the riparian communities and save cashof many households settled plant near the wetland. Nevertheless, this study also revealed that some near the wetland resources are declining due to population pressure, encroachment near the wetlands, deforestation and over-exploitation of the wetland habitats for creation of many fishery ponds within the wetland and cultivation practices on wetland edges. The socio-economic condition, unemployment, easy accessibility and lack of alternativesources are plausibly themajor factors, whichare leading to the decline and depletion of the wetland resources. Therefore, appropriate management strategies are required for the conservation and the sustainable utilization of these plant resources.

The productivity of a crop is result of genetic make-up (heredity) and environment (climate) and their interaction. The components of crop environment or climate or microclimate are light, temperature, air compositions and the nature growing medium or soil. In open fields, only manipulation of nature of the soil or root medium by tillage, irrigation and fertilizer application is possible. Under greenhouse structure control of any one or more of the components of the micro climate is manipulated.

Light

The visible light of the solar radiation is a source of energy for plants. Light energy, carbon dioxide (Co2) and water all enter in to the process of photosynthesis through which carbohydrates are formed. The production of carbohydrates from carbon dioxide and water in the presence of chlorophyll, using light energy is responsible for plant growth and reproduction. The rate of photosynthesis is governed by available fertilizer elements, water, carbon dioxide, light and temperature. The photosynthesis reaction can be represented as follows

For plant analysis to be meaningful, collection of particular plant part(tissue) at the right stage of growth is very important. A research worker in soil fertility is generally interested in finding out the nutrient uptake by crop at different stages. The stage at which nutrient uptake is to be studied will vary with different crops. One may also be interested in collecting a plant sample when some abnormality is observed in the plants and a nutrient deficiency or toxicity is suspected. In such cases no time should be lost in collecting the sample otherwise secondary reactions that often take place in the mineral content of plants will conceal the real cause of deficiency or toxicity. In such cases only the affected plants are to be sampled as compared to general sampling technique described below.

For research projects each plot receiving differential fertilizer or some other treatment is to be sampled separately. For general advisory purposes one sample would be good enough for the entire field (not exceeding 2 hectares). One should, however, take care that the sample comes from a field or a part there of having a uniform stand. If a field is patchy and differences in stand and plant growth are obvious, each such area should be sampled separately.

Introduction

Plant analysis can be defined as the quantitative determination of the concentration of an element or extractable fraction of an element in a sample from a particular part or portion of a crop. From the nutritional standpoint, plant analysis is based on the principle that the concentration of a nutrient within the plant is an integral value of all the factors that have interacted to it. Plant analysis is used as an index of available nutrient element supply. Plant growth or yield are compared with the elemental concentrations contained in the dry matter of the entire plant or plant structures such as leaves, petioles, fruit or grain sampled at different times during their development. Plant analysis gives the overall picture of the nutrient levels within the plant at the time the nutrient was taken. The use of plant analysis is based on the principle that the nutrient level present is as a result of all factors affecting the growth of the plant.

Plant analysis involves the determination of nutrient concentration in diagnostic plant part(s) sampled at recommended growth stage(s) of the crop. In a way plant analysis complements soil analysis. There are reliable sampling criteria and procedures for most of the world’s commercial crops.

Importance

Plant analysis assesses nutrient uptake while soil testing predicts nutrient availability. The two tests are complementary to each other as crop management tools. Plant analysis will detect unseen hidden hunger and confirm visual deficiency symptoms. Toxic levels may also be detected. If it is done early, plant analysis will allow a corrective fertilizer application in the same season.

Summary

Plant cell culture systems produce myriads of valuable compounds which cannot be synthesized by microbial cells or chemical synthesis. Many of the drugs sold today are simple synthetic modifications or copies of the naturally obtained substances. The burgeoning demand for secondary metabolites in recent years as therapeutic agent has resulted in a great interest, in secondary metabolism, and particularly in the possibility to alter the production of bioactive compounds using cell culture technology. Various strategies such as cell line selection, use of precursors and elicitors, mass culture in bioreactors and others have proved effective in enhancing the production of these metabolites so as to meet the market demand but space for improvement still exists. In order to stretch the boundary, recent advances, new directions and opportunities in plant cell based processes are being examined in this section.

Summary

Plant tissue culture is the technique of growing plant cells, tissues or organs in isolated from the mother plant on artificial media. It has opened up unprecedented opportunities in many areas of basic and applied biological research. This includes techniques and methods used to research into many botanical disciplines and has several practical objectives of plant tissue culture and micropropagation for sustainable development. Indeed, the landmark publication by Gottlieb Haberlandt who is arguably referred to as the “Father of Tissue Culture”, is often cited as the origin and emergence of plant tissue culture and its subsequent applications. Success of biotechnological approaches is dependent on regeneration of intact plants following genetic modification, generally by micropropagation. Potential uses of plant tissue culture in biotechnology to further our understanding of plant physiology, how plants function, resolution and expression of use full characters form in vitro to in vivo condition. Agro bacterium-mediated genetic transformation along with other modes of transformation such as microprojectile bombardment has been accomplished mainly through plant tissue culture.

Introduction

Among the various propagation techniques, plant propagation through tissue culture has gained immense importance over the years and for a few crops it has become the major method of propagation. Plant tissue culture can be defined as the ability to grow and maintain the plant organs viz., embryo, shoot, root, anther, flower and their respective tissue in a culture media. While micropropagation can be defined as the development of a new plant using plant tissue culture techniques. These plants are grown under in-vitro conditions, i.e., culturing the plant in a vessel (e.g., test tube) under a controlled condition. Since, the whole process is a lab-based work, there are chances of contamination, thus, the growing condition needs to be aseptic (growing without microbial contamination).

Media Preparation

Before mixing medium, determine the quantity of each medium and the amount of plant growth regulator needed for each experiment. Make slightly more medium than is required. A media check off list is helpful in making medium to keep track of ingredients.

Remove media stock solutions and plant growth regulator stock solutions to be used from the refrigerator. Make up fresh stock solutions if adequate quantities are not available or if solutions have precipitated.

Plant growth regulators stock solutions are prepared biweekly and stored at 4º C in the dark. Cytokinins (thidiazuron and BA) are dissolved in approximately KOH prior the addition of deionized water. Gibberellins and auxins (2,4-D, IBA and NAA) are dissolved in 100% ethanol prior the addition of water.

Fill the volumetric flasks upto ½ to ¾ with deionized water. Add proper a of stock solution and plant growth regulators and mark each item on the check off list. Add 30 g/l sucrose, stir until dissolved. Make up the volume of the flask with deionized water and later por into a larger beaker.

Introduction

One of the biggest problems emerging for the survival of the people in the future is food security. To feed an anticipated nine billion people by 2050, the globe would need to produce roughly twice as much food as it does today. In order to properly accomplish this aim, food production must be elevated sustainably on currently available arable land while addressing the problems brought on by climate change. Over the past five decades, crop yields have steadily increased due to crop breeding programs and better management practices. The rate of yield improvement has, however, reached a plateau (Grassini et al. 2013). The noncompounding yearly yield growth of 0.9% for wheat at the present time, world output rises by 38%, is quite small, and is unable to fulfill expected demand levels by 2050. Therefore, the most beneficial course of action is to investigate more effective, higher-yielding crops (Ray et al. 2013).

Historically, the genetic improvement of wheat has been accomplished by sexual hybridization between related species, giving rise to a large number of cultivars with excellent agronomic performance and high yields. The main methodology for improving cereal crops is still traditional plant breeding, sometimes in combination with traditional cytogenetic methods (Salina et al. 2015). Cereal crops immediately became the top targets for genetic modification due to the worldwide importance of cereal grains in human diets. Over the past 10 years, wheat genetic technology for processing has advanced quickly.

Plant tissue culture is the cultivation of plant cells, tissues, or organs on specially formulated nutrients media under the right environmental conditions. A whole plant can be regenerated from a single cell. The history of plant tissue culture has been long and started with Haberlandt in 1902 when he proposed the culture of single plant cells, his experimentation never resulted in success. White (1943) became successful in culturing plant cells for the first time, however, Morel (1960) at the Institut National Recherche Agronomique in Versailles near Paris was first to demonstrate meristem culture technology. He successfully introduced tissue culture technology to the orchid industry and produced disease-free plants of grape. By the 1970s, plant tissue culture had become a formidable technology. Plant tissue culture technique has been around for more than 30 years and considered as an important straightforward technology for the production of disease-free high quality planting material and the rapid production of many uniform plants. Its application requires a sterile workplace, nursery, greenhouse and trained manpower.

Plant tissue culturing techniques have been especially important in the agricultural community over the past many years. During this time, plant tissue culture has effectively moved from the confines of small laboratories and has taken its place among broad scale techniques employed by agricultural industry.

Tissue culture is a technique used to culture cells, tissues, organs or whole plant under controlled environmental conditions that in vitro condition is known as plant tissue culture. Plant tissue culture exploits the totipotency nature of plant cells, ability to develop in to complete plantlet.

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
The process of culturing plant cells, tissues, organs and protoplast on artificial media under controlled and aseptic circumstances is known as ‘Plant Tissue Culture’. It is predicated on the idea of “Cellular totipotency”, which describes a cell’s capacity to develop into a whole plant in the right cultural environment. This characteristic of cells has broad applications in the manipulation of plant cells to enable fast plant multiplication and the in-vitro regeneration of whole plants following the genetic transformation. Around a century ago, the idea of tissue culture was first put forth, to grow entire plants in vitro from somatic cells (Haberlandt, 1902). The tissue culture system has advanced with the identification of different concentration, ratios of auxin and cytokinin, which are critical for the regeneration of adventitious roots and shoots (Skoog and Miller, 1957). Appropriate explant, or plant organ removed, including cotyledons, leaves, roots, shoot apices, nodal segments, hypocotyls, anthers, embryos, and seeds. The explants are thoroughly cleaned with sterile double-distilled water, surface sterilized with a disinfectant such as sodium hypochlorite or mercuric chloride and aseptically cultured in an artificial medium in test tubes/ flasks, jars, and Petri dishes. Several factors influence a plant’s capacity to regenerate, such as the presence of a plant growth regulator (Gerdakaneh et al., 2020), the explant type (Dhar and Joshi, 2005) and the basic medium’s composition (Sundararajan et al., 2017).
The most widely used media include major elements, carbohydrates, growth regulators (auxins, cytokinins etc.), microelements, vitamins, amino acids and several media compositions have been created specifically for plant tissue culture (Atkinson et al. 2012). While 2,4-dichloro phenoxy acetic acid (2,4-D) at concentrations of 0.5-4.0 mg/L typically induces callus, or a homogeneous mass of undifferentiated cells, auxins at concentrations ranging from 0.1 to 5.0 mg/L, favour rooting by cell elongation. Similarly, cytokinin’s induce fast cell division and the growth of shoot buds/shoots. Agaragar, agarose, and GelriteTM are examples of chemically inert, powdered gelling agents added to the culture medium to solidify it before it is autoclaved. After the medium has been added to the culture containers, they are autoclaved. Explants in the culture vessels are injected under aseptic circumstances in a laminar air-flow cabinet equipped with high-efficiency particulate air (HEPA) filters (pore size 0.2–0.3 mm). Explants that are placed in the right growth medium under the right circumstances dedifferentiate, or mature cells that have increased DNA/ RNA and protein production return to their meristematic state. Regrowth in the agar-gelled medium results in an amorphous mass of cells known as ‘callus’. Suspension culture, a method of cultivating cells in a liquid medium, produces a suspension of individual cells. Every three to four weeks, the developing tissue must be sub-cultured onto a new medium because an increase in cells or calli causes the medium to run out. In the end, the organogenesis pathway is utilized to create ordered structures like roots, branches, flower buds, etc. from these cells and tissues. The synthesis, preservation, and application of

Meristematic Tissues

These are composed of cells that are in state of division or retain the power of dividing. These cells are essentially alike and isodiametric. They may be spherical, oval or polygonal and their walls thin and homogeneous; the protoplasm in them abundant and active with large nuclei, and the vacuoles small or absent.

Classification of Meristems

Meristems are classified in different ways on the basis of certain factors.

A) According to their origin and development

1. Promeristem or primordial meristem

2. Primary meristem and

3. Secondary meristem.

Key Points

• It might be necessary to administer oxygen or perform artificial respiration. Giving tranquilizers is not a good idea.

• Precaution: It's crucial to always follow the label's instructions on how to use pesticides since they provide information on how to do so. Warnings against using on unapproved species or in testing conditions must also be included on labels. A maximum of care should be made to prevent drift or drainage to neighboring fields, pastures, ponds, streams, or other properties outside the treatment area. In no case should amounts larger than those specifically recommended be utilized.

Introduction

It has been determined that a number of plant species are poisonous to animals. These plants not only posed a health risk to humans and animals, but they also significantly reduced cattle productivity. Many plants contain chemicals that can be dangerous or even lethal to animals. The degree of toxicity varies based on the animal species, the quantity of plant material ingested, and the particular poisonous compounds in question. In general, there are two categories of dangerous plants: those that are known to be toxic and contain toxic compounds, and those that are normally non-toxic but can turn hazardous in certain circumstances. Here are some typical plant toxicities, along with information on managing toxicity and clinical signs.

5.1 Plant Breeder’s Rights (PBR) and Protection of Plant Varieties and Farmers’ Rights (PPV&FR) Act
5.1.1 Introduction
In the context of biotechnology and agricultural innovation, Plant Breeder’s Rights (PBRs) offer exclusive intellectual property protection to individuals or organizations that breed, develop, or discover new plant varieties. These rights ensure economic incentives for plant breeders while promoting innovation in seed development and agricultural productivity. In India, the unique and inclusive Protection of Plant Varieties and Farmers’ Rights Act (PPV&FR), 2001, balances breeders’ rights with recognition of farmers’ contributions, making it one of the most progressive legislations in the world.
5.1.2 Concept of Plant Breeder’s Rights (PBR)
Plant Breeder’s Rights refer to the exclusive rights granted to plant breeders to produce, sell, market, distribute, import or export their registered plant varieties. These rights are sui generis, meaning they are a special form of IP distinct from patents, developed to cater specifically to plant varieties.
Key Objectives of PBR
• Encourage investment in plant breeding and biotechnology.
• Ensure access to improved crop varieties.
• Prevent unauthorized commercial exploitation of novel varieties.
• Facilitate biodiversity conservation through formal registration systems.