eChapters

ABSTRACT

The multi-sensor airborne geophysical survey was carried out over Nalgonda district, Andhra Pradesh, during the field season 2002-03, with an objective to locate potential areas for mineral exploration. The area comprises older metamorphic rocks of Peninsular Gneissic Complex of Archaeanage represented by gneisses, granite, amphibolite and banded magnetite quartzite. Leucocratic intrusive granite, extending over a length of about 40 km around Suryapet in NNW-SSE direction exhibits intense fracturing. Two sets of vertical to sub-vertical fractures have been noted in granite. Yellow stains observed in granite near fracture planes reveal the presence of secondary radioactive mineral encrustation. The ground magnetic and spectrometric observations indicated that the uranium mineralization is confined to these fractures in granite. Petrographic, ore-microscopic and scanning electron microscopic studies revealed evidences of metamict alteration surrounding monazite/apatite inclusions within biotite, and magnetite grains in granite. These studies also indicated evidences of discrete grains of uraninite / coffenite / pitchblende (containing Thand Ce). Chemical analysis of the rock samples (by NAA) collected from anomalous zone reveal high uranium concentration of the order of 1200 ppm, marked by high values of total Rare Earth Element concentration (800 ppm).

The leucogranites are characterized by high LREE and a negative Eu anomaly suggesting plagioclase fractionation. An ultramafic enclave near village Kutubsahipur shows highest concentration of both uranium (1240 ppm) and REE (890 ppm) analyzed by BARC and NAA Lab, GSI.

ABSTRACT

Frankia is a gram-positive, microaerophilic, actinomycete that enters into symbiotic associations with a number of woody dicotyledonous plants called actinorhizal plants. Frankia can fix atmospheric nitrogen through such associations. The actinorhizal-Frankia association is beneficial for the environment, since it increases the fertility of soils. Extensive studies on the biochemical and molecular aspects helped in the classification of Frankia. PCR-RFLP studies of the 16S rRNA and the nif regions of Frankia revealed considerable information regarding the diversity of Frankia strains, isolated from different parts of the world. The completion of three whole genomes of Frankia has bolstered the study of the molecular organization of the genome. Comparative genomics study of the three complete genomes revealed that, the strains reflected the biogeographical history of the host plants they infected. Striking similarity in the codon usage patterns highlights the fact that they may have evolved as a unit. In the present review we highlighted the taxonomy of Frankia and the host plants, the Frankia-actinorhizal association, regulation of infectivity, biochemical and molecular characterization, phylogeny and genomics of Frankia and stressed the need for metabolomic study of the Frankia-actinorhizal association.

ABSTRACT

Serious infections caused by bacteria that have become resistant to commonly used antibiotics have become a major global healthcare problem in the 21^st century. Antibiotic resistance, initially a problem of the hospital setting associated with an increased number of hospitals acquired infections usually in critically ill and immuno-suppressed patients, has now extended into the community causing severe infections difficult to diagnose and treat. In hospitals, most common resistant bacteria include methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci and gram-negative rods including Enterobacteriaceae and Pseudomonas aeruginosa. Vancomycin intermediate and resistant S. aureus, represent a new treatment challenge. In the community, penicillin and macrolide-resistant pneumococci developed several decades ago and are now present all over the world. More recently, communityacquired methicillin-resistant S. aureus has become a problem in several countries causing skin infections but also severe diseases. The molecular mechanisms by which bacteria have become resistant to antibiotics are diverse and complex. Bacteria have developed resistance to all different classes of antibiotics discovered to date. The most frequent type of resistance is acquired and transmitted horizontally via the conjugation of a plasmid. In recent times new mechanisms of resistance have resulted in the simultaneous development of resistance to several antibiotic classes creating very dangerous multidrug-resistant (MDR) bacterial strains, some also known as ‘‘superbugs’’. In many cases the use of antibiotics is unnecessary or questionable. The indiscriminate and inappropriate use of antibiotics in outpatient clinics, hospitalized patients and in the food industry is the single largest factor leading to

ABSTRACT

The continuing depletion of the stratospheric ozone layer mainly due to release of man-made atmospheric pollutants such as chlorofluorocarbons (CFCs) have resulted in the increase of solar ultraviolet-B radiation (UV-B; 280-315 nm) reaching to the Earth’s surface. UV-B is a small (less than 1% of total energy) but highly active component of the solar spectrum that can penetrate to biologically significant depths in lakes, ponds, rivers and oceans. It has been demonstrated that UV-B can cause wide ranging effects including mutagenesis, destruction of proteins/enzymes, inhibition of a number of vital metabolic processes and formation of thymine dimers in DNA leading to death of a number of organisms. A number of microbes including strict photoautotrophs always grow in lighted habitats and thereby always sense and face the stress of solar UV-B radiation. Similarly, humans are also exposed to varying levels of UV-B radiation depending upon the altitudes and latitudes of the location. A wide ranging effect of UV-B such as cataracts, skin cancer, immunosupression etc. has been reported in human. Damaging effects of increased UV-B radiation has also been shown in a number of microbes. These effects include inhibition of growth and survival, decrease in photosynthetic pigments, loss of PS II and RuBISCO activity, inactivation of nitrogenase enzyme and inhibitory effects on several other metabolic processes. Among photosynthetic pigments, phycobiliproteins seem to be the primary target of action since phycobilisomes are completely disintegrated after UV-B exposure. In this contribution an attempt has been made to put in perspective the main effects of UV-B radiation on microbes and human health.

ABSTRACT

Heavy metal contaminated soils are biologically inactive and extremely poor in soil fertility status. These metals are highly resistant and cannot be easily degraded by either chemical reactions or amendments. Microorganisms are biological softwares capable of degrading the metals efficiently. Among microorganisms, mycorrhizas are known to circumvent metal toxicities through a series of processes that assist in immobilization in biological systems which converts toxic metals into less or nontoxic forms. There are five mechanisms such as biosorption, solubilization, hyphal transport, detoxification and dilution are involved in promoting tolerance of host plants to heavy metal toxicities. These processes are strategies that can be exploited to phytoremediate the contaminated soils. The phytoremediation strategies encompass tolerance of mycorrhizal fungi, phytostabilization (preventing entry of heavy metals into plants), phytoextraction (luxuriant uptake and transport to shoots) and intercropping or crop rotation with mycorrhizal dependent hyperaccumulators or non-hyperaccumulators. Despite mycorrhizas are natural, the indigenous population is not sufficient enough to induce tolerance of host plants to abiotic and biotic stress conditions. This necessitates adopting recommended agronomic practices to promote mycorrhizal colonization in natural agro-ecosystem or to derive full benefits from phytoremediation strategies. Thus mycorrhizas are important naturally occurring microorganisms capable of withstanding highly contaminated metal sites and offer protection against phytotoxicities caused by heavy metals.

ABSTRACT

Plasmids are circular, double-stranded DNA molecules which replicate autonomously from the chromosome of the host organism. Plasmids are found in many bacterial species and often confer upon the host cell phenotypic characteristics such as resistance to antibiotics, resistance to heavy metals, ability to produce particular protiens, conjugative (mating) ability and ability to degrade xenobiotc compounds. One of these degradative plasmids may encode a complete catabolic pathway (such as toluene to pyruvate, formate and acetaldehyde) or partial degradative steps, such as octane to octanol. In this later case, the remainder of the pathway may be encoded by chromosomal genes, by genes of other plasmid present in the host, or by other microorganisms. Many degradative plasmids possess a broad host range, especially among the gram-negative bacterial species.

There are different mechanisms which may be involved in the evolution of aromatic pathways in general and in the process of adaptation of microorganisms to xenobiotic substrates in particular. Gene transfer, the occurrence of plasmids in bacteria in the natural environment is an important pool of genetic information residing on plasmid vehicles may flow among indigenous microorganisms. Since many examples of self-transmissible plasmids that carry genes for degradation of aromatic or other xenobiotic compounds are known, and their role in spreading these genes to other microorganisms is well characterized. Early, studies of TOL, NAH, and SAL plasmids revealed strong homologies among these plasmids in regions both inside and outside the catabolic gene clusters. Plasmids, pJP4, pAC25, and PSS50, catabolic plasmids carrying genes for chloroaromatic degradation, have strongly homologous plasmid backbone that determine replication and transfer function. Furthermore, the chlorobenzoate plasmid p51 was also shown to be homologous to pJP4 outside regions of the catabolic genes. These observations suggest that a few common self-transmissible ancestor replicons (includes origin of replication and replication gene) may have been involved in acquisition and spread of different catabolic genes. DNA

ABSTRACT

An understanding of the metabolism of heavy metals facilitates biotechnological use of these functions just recently, although heavy metals comprise the major part of the elements in the periodic table. Due to their ability to form complex compounds, some heavy metals are essential trace elements. In any case, essential or not, most heavy metals are toxic at higher concentration. Heavy metal pollution from last few decades has led to an increased environmental concern. Heavy metal like cadmium is a non-essential toxic heavy metal and is also being increased in soil during last few years posing a serious threat to environment and human health worldwide. Therefore, remediation strategies of the contaminated sites are great concerns and can be achieved by a number of different physico-chemical and biological means.

ABSTRACT

Industrial and agricultural activities and the disposal of waste products have resulted in the contamination of many terrestrial environments. Industrial effluents often contain considerable amount of potentially toxic compounds such as Cd, As, Pb and Hg. Due to their small size, which provides a large contact area that can interact with the surrounding environment, microorganisms are the first biota showing the impact of toxic compounds. Microorganisms being in intimate contact with the soil environment are considered to be the best indicators of soil pollution. In general, they are very sensitive to low concentrations of contaminants and provide a rapid response to soil perturbation. Rhizobium spp. are ubiquitous gram-negative soil bacteria that have a profound scientific and agronomic significance due to their ability to establish nitrogen-fixing symbiosis with legumes, which is of major importance to the maintenance of soil fertility and to feed livestock and populations in many developing countries. Therefore, the present work aimed to evaluate the impact of heavy metals on Rhizobium populations isolated from different locations contaminated with heavy metals. In order to reach this goal some physicochemical parameters that influence metal bioavailability were determined, as well as their concentrations in soil. Cadmium tolerance of Rhizobium isolates was screened in artificial media and the highest cadmium concentration allowing growth was defined as the maximum resistance level (MRL). The heavy metal tolerance variability found among Rhizobium isolates, suggests that beyond basic metabolic machinery also exist variations that allow isolates to display distinct tolerance responses to Cd stress. To better understand Rhizobium cadmium tolerance, wall-bound and intracellular Cd was determined. Sensitive and moderately tolerant isolates showed low intracellular Cd amounts when compared to extremely tolerant isolates, which were able to withstand intracellular Cd concentrations 40 times higher. Lipopolysaccharides (LPS) bind cations unspecifically, so they constitute the initial defence mechanism to heavy metals since they sequestrate cations and therefore prevent their entry into the cell. LPS quantity varied significantly between isolates. LPS increases were observed in most sensitive, moderately tolerant and tolerant isolates, evidencing an attempt of cells to adjust to cadmium entry. These results may be of great importance for understanding how Rhizobium persists in contaminated soils. They also provide important data to the development of bioremediation processes and to the improvement of soil productivity.

ABSTRACT

The world ocean with a coastline of approximately 3,50,000 km is the largest ecosystem on earth. Marine organisms are the major, sustaining components of ecosystem processes and are responsible for biogeochemical reactions that drive our climate changes. Despite this, many marine organisms are poorly described and little is known of broad spatial and temporal scale trends in their abundance and distribution. With new molecular and analytical techniques we can advance our knowledge of marine biodiversity at the species level to understand how marine biodiversity supports ecosystem structure, dynamics and resilience. We can then interpret environmental, ecological and evolutionary processes controlling and structuring marine ecosystem biodiversity. In biotechnology applications marine microbes represent a potential source for numerous commercially important biomolecules and their bioremediation capabilities are also remarkable. They also play a crucial role in decomposition of organic matter and cycling of nutrients. Microbes also serve as food for some bottom-living organisms. Understanding and preserving biodiversity was one of the most important global challenges for the past 20 years and will continue to be an important scientific issue into the new millennium. Our knowledge of marine microbial diversity has, however, been severely limited by relying on microorganisms that have been cultured which are less than 1%.

ABSTRACT

Endophytes are microorganisms that live inside the host plant tissues without causing symptoms of disease. There has been a crescent interest in the prospecting of these microorganisms as sources of novel and bioactive natural products. In fact, a significant number of interesting natural products have been reported in the last years. In this chapter some aspects regarding the endophytes are detailed: the relationship with their host plants, isolation, preservation, taxonomy, cultivation, frequently isolated strains, tropical endophytes, novel structures isolated and also their biological activities.

Key words: natural product, endophytic fungi, plant fungus interaction, secondary metabolites.

ABSTRACT

Soil borne fungal pathogen causing root diseases of vegetables are difficult to control by chemical methods. The existing biocontrol bacteria are not very effective in control of soil borne fungal pathogens. As such very less number of bacterial biopesticide is available commercially to control the fungal pathogens. Increasing concerns in recent years about human health and environmental problems due to chemical pesticides, couple with increasing pest resistant to chemical, have led to a reappraisal of our approach to control pest and disease in crops. The trends now are towards a management concept in which the best combination of tools is used to manage plant diseases below economic thresholds with minimal risk to human health and environment. The modern management concept includes extensive use of biocontrol agents which are harmless to non-target organisms. Development of effective technology for control of pest and disease by using microbial resources of a region to enhance the quality of ecosystem is a potential alternative approach to chemical control of plant diseases. In such strategy, search for microorganisms and biological system is an important prerequisite prior to development and practical exploitation of biocontrol. Rhizosphere microorganisms are shown to be perspective candidate for biocontrol agent as they provide the front line of defense for plant roots against the attack by various plant pathogens causing the most diseases of vegetables and crops. Chemical control of soil-borne fungal pathogen had been the subject of growing concern because of their adverse effect on the environment and emergence of chemical pesticide resistant to plant pathogen, moreover, soil borne pathogens are very difficult to control. Crop rotation, breeding for resistant plant varieties and application of pesticides are insufficient to control root diseases of important plants because they survive as sclerotia and has got wide host range.

ABSTRACT

Plant growth promoting (PGP) bacteria play an immense role in promoting crop growth mainly by direct mechanisms such as N_2-fixation, phosphate solubilization and phytohormone production and indirectly may help to suppress pathogens or contribute to enhance absorption of nutrients leading to increase yields. The N₂fixers are grouped based on their niche adaptation as endophytic nodular symbionts (Rhizobium spp.), free living (Azotobacter spp.), associative symbionts (Azospirillum spp.) and endophytic non-nodular symbionts (Azoarcus and Gluconacetobacter diazotrophicus). In the recent past, phosphate solubilizing and phytohormone synthesizing bacterial communities also gained momentum.

ABSTRACT

Methylotrophy is a phenomenon by which microorganisms derive energy, and in many cases, cell carbon from reduced molecules that have no C–C bond (also called C1compounds). This gains significance because of its unique mode of nutrition, the ability to utilize more than one metabolic pathway and its vast distribution among various groups of bacteria and yeasts. It is one of the earliest modes of energy metabolism present in microorganisms, through which microorganisms are able to utilize a wide range of compounds, which are considered unsuitable for most forms of life. In the present chapter we focus on various aspects of methylotrophy, and the application of methylotrophic bacteria in a spectrum of biological activities and commercial possibilities.

ABSTRACT

Biotransformation is an essential step in the elimination of drugs and reduction of their toxicity. The use of fungi in biotransformation reactions can provide information about the metabolism of new drugs and facilitate the attainment of necessary amount of metabolites for toxicological and pharmacological evaluation. Biotransformation studies require the right choice of the microorganisms and also of the culture conditions (pH, substrate, oxygen, salts, temperature, etc). The optimization of these procedures is essential to achieve good results. The fundamental aspects of drug biotransformation by fungi are discussed here as well as some applications showing that many fungi are used in biotransformation studies, mainly Cunninghamella species. Several species mimic the mammalian metabolism producing the same metabolites observed in humans.

ABSTRACT

A number of secondary metabolites are produced by cyanobacteria involving the non-ribosomal peptide biosynthetic pathway. Some of these compounds have potential biotechnological applications, whereas others (the cyanotoxins) are undesired due to their toxicity to animals. The occurrence of toxic cyanobacterial blooms in freshwater bodies that are used as source for human consumption, recreation and irrigation are frequent nowadays due to increasing artificial eutrophication of these environments. It is important to monitor toxin-producing strains in order to prevent the adverse effects caused by their toxins to human and animal health. Thus, the development of rapid and sensitive methods for the detection of cyanotoxins is important as they could be useful to monitor water supplies and contribute for watershed monitoring strategies. Current methods include cell counting, microscopic identification and toxin analysis by biochemical and analytical techniques. However, in recent years, molecular biology techniques, such as polymerase chain reaction (PCR), have been used to amplify specific regions within biosynthetic genes for rapid and sensitive identification of toxic cyanobacteria. The aim of this chapter is to provide an update of cyanotoxin biosynthesis and discuss the advances and limitations of molecular biology regarding its application as indicative of cyanotoxin production.

ABSTRACT

Bioinformatics is a critical tool in designing the molecular biology experiments. The xeroxing of specific part of genome needs correct primer designing. Primer designing has central role in molecular biology right from cloning, sequencing, gene expression profiling by real time PCR, genotyping by RFLP, SNP analysis by real time PCR and auto sequencer. In this chapter we are describing how to proceed for experimental designing right from data mining from GENEBANK to designing of primers for various common molecular genetic analysis work. The write up also describes in silico evaluation of primers, various off line commercial and non-commercial software, on line free software and trouble shootings in primer designing, The different strategy described in context of primer designing would be of immense use of researchers.

The effects of global warming and/or climate change are of concern both for the environment and human life. Evidence of observed climate change includes the instrumental temperature record, rising sea levels, and decreased snow cover in the Northern Hemisphere. According to the IntergovernmentalPanel for Climate Change (IPCC) Fourth Assessment Report, “most of the observed increase in global average temperatures since the mid-20th century is very likely due to the observed increase in [human greenhousegas] concentrations. It is predicted that futureclimate changes will include further globalwarming (i.e., an upward trendin global mean temperature), sea level rise, and a probable increase in the frequency of some extreme weather events. Ecosystems are seen as being particularly vulnerable to climate change. Human systems are seen as being variable in their capacity toadapt to future climate change. To reduce the risk of large changes in future climate, many countries have implemented policies designed to reducetheir emissions of greenhouse gas.

Phytochemicals are chemical compounds that occur naturally in plants (phyto means “plant” in Greek). Some are responsible for color and other organoleptic properties, such as the deep purple of blueberries and the smell of garlic. The term is generally used to refer to those chemicals that may have biological significance, for example antioxidants, but are not established as essential nutrients. Scientists estimate that there may be as many as 10,000 different phytochemicals having the potential to affect diseases such as cancer, stroke or metabolic syndrome. The potential of the phytochemicals have large scale pharmacological and biological activities such as antioxidant constituents (hydrolysable tannins, phenolic acid and flavonoids) of the plant materials for the care of health and protection from coronary heart diseases, cancer, anti-carcinogenic and anti-mutagenic effects. Varieties of herbaceous vegetables are protective against various diseases, particularly cardiovascular diseases. These herbaceous plants and species are harmless sources for obtaining natural antioxidants. Antioxidant constituents can delay or inhibit the oxidation of lipids and other compounds by inhibiting the propagation of oxidation chain reaction (Hussain et al., 2011)

Primarily, antioxidant effect is due to phenolic compounds such as phenolic acid, flavonoids and phenolic diterpenes and their mode of action for antioxidant compounds is due to its redox reaction properties which can absorb and neutralize free radicals by quenching singlet and triplet oxygen. Vitamins are organic substances necessary for metabolism. Human diet does not always contain the required amount of vitamins for the normal growth and maintenance of the body function and as such cannot produce enough quantity for their body metabolism, so it can be obtained from fruits, vegetables and foods. Deficiency of vitamins can cause serious human health diseases and sometimes, very small concentrations are required for maintenance of good human health.

Equipment

1. Adjustable pipette.
2. 20-ml glass beaker.
3. Atomic Absorption Spectrophotometer.

In chemistry, and especially in biochemistry, a fatty acid is a carboxylic acid with a long aliphatic tail (chain), which is either saturated or unsaturated. Most naturally occurring fatty acids have a chain of an even number of carbon atoms, from 4 to 28. Fatty acids are usually derived from triglycerides or phospholipids. When they are not attached to other molecules, they are known as “free” fatty acids. Fatty acids are important sources of fuel because, when metabolized, they yield large quantities of ATP. Many cell types can use either glucose or fatty acids for this purpose.

Nitrogen is a nutrient and occurs in many forms including ammonia, organic, nitrate and nitriteeach of which may be tested for in a variety of ways. Under normal circumstances, the nitrite form of nitrogen will not be present in large quantities due to its rapid oxidation orconversion to nitrate. The presence of large concentrationsof ammonia in a stream or lake can create a large oxygen demand. This demand is caused by the conversion of ammonia to nitrate. High concentrations of nitrate in waste water treatment plant effluent can cause algae to grow in large quantities.

Current status

For many years, the protein contentof foods has been determined on the basis of total nitrogen content, while the Kjeldahl (or similar) method has beenalmost universally applied to determine nitrogen content (AOAC, 2000).Nitrogen content is then multiplied by a factor to arrive at protein content. This approach is based on two assumptions: thatdietary carbohydratesand fats do not contain nitrogen, and thatnearly allof the nitrogen in the dietis present as amino acids in proteins. On the basis of early determinations, the average nitrogen (N) content of proteins was found to be about 16 per cent, which led to use of the calculation Nx6.25(1/0.16 = 6.25) to convertnitrogen contentinto protein content.

Carbohydrates are widely prevalent in the plant kingdom, comprising the mono, di-, oligo-, and polysaccharides. The common monosaccharides are glucose, fructose, galactose, ribose etc. The disaccharides, i.e., the combination of two monosaccharides include sucrose, lactose and maltose. Starch and cellulose are polysaccharides consisting of many monosaccharide residues. Cellulose is the most abundant organic compound on this planet since it forms part of the cell wall in plants.

Aldehydes (–CHO) and ketones (= CO) are active groups in carbohydrates. Carbohydrates contain many hydroxyl groups as well. The number of hydroxyl groups varies with the number of carbon atoms. Monosaccharides contain the free aldehyde or ketone group. Some disaccharides have the free aldehyde group (maltose) and some do not have the free ones (sucrose). The polysaccharides, starch and cellulose, are polymers of monosaccharides linked through the active groups. The chemical properties of saccharides vary depending upon the number of hydroxyl groups and the presence or absence of –CHO/= CO groups. These variations are the basis in the development of colour reactions to identify the saccharides. Some simple tests used to identify the presence/absence of certain saccharides are listed below:

Plants are exposed to various stresses such as temperature, water, oxygen, salts, nutrition etc, during its growth under dry land condition. Various simple tests have been developed by plant scientist for evaluation of different cultivars against these stresses. These tests are simple to perform in the laboratory with fewer instruments that are commonly available in most of the laboratories in developing countries. These tests have significance in plant water relations and cultivars reliably screened for various stresses. The crops upon which these tests have been performed here are used by the proponents of the tests. These tests are not advised to be performed on crops other than those used by the authors. It probably needs standardization for other crops. The following test has been explained for the benefit of the readers.

Hydroponics is a subset of hydroculture and is a method of growing plants using mineralnutrient solutions, in water, without soil. Terrestrial plants may be grown withtheir roots in the mineralnutrient solution only or in an inert medium, suchas perlite, gravel,mineral wool, expanded claypebbles or coconut husk

The techniques on hydroponics are widely used in plant sciences to elucidate several problemson nutrition. Primarily, this has been not only used to evaluate deficiency symptoms of an essential element but also used to assess waters stress effects in crop plants. There has been a considerable controversy whether such responses paralled with field responses. Though these techniques have their limitations, it is certain that it has its relevance in identifying certain genotypes relatively tolerant to some stresses such as water, O₂ and nutrition. Without entering to such debatable issue, it can be said thatwherenecessity demands, such techniques would proveuseful in demonstration purposes and evaluation of macronutrient deficiencies in tomato and maize. Thegrowing of plants in nutrient solution is called as water culture or hydroponics. These experiments in nutrient solutions are carried out to study the mineral nutrient relation of plants. By elements required for plant growth, specific deficiency symptoms can be produced in water culture experiments. Thesuccess of these experiments of producing deficiency symptoms depends upon the use of homogenous biological material and purity of culture solution in respect of different plant nutrients. Hydroponics is a subset of soilless culture. Many types of soilless culture do not use the mineral nutrient solutions required for hydroponics.

Plants that are not traditionally grown in a climate would be possible to grow usinga controlled environment systemlike hydroponics. NASA has also looked to utilize hydroponics in the space program. Ray Wheeler, plant physiologist at Kennedy Space Center’s Space Life Science Lab, believes thathydroponics will create advances within space travel. He terms this as a bioregenerative life support system.

Growth and Development are the most fundamental and conspicuous characteristics of all living organisms. According to dictionary, growth is the advancement towards maturity and development is a gradual increase in size. The plant physiological definition of growth is ‘an irreversible increase in mass, weight or volume of a living organism, organ or cell.

Tissue culture refers to the collection of techniques used to maintain or grow of cells, tissues and organs under controlled environment. The German botanist G. Haberlandt (1854-1945) originated the concept of cell culture in which he described his pioneering experiments with isolated plants cells and developed a concept of totipotency (potential of a cell to reproduce entire plant and all of its characteristics). He is considered as father of tissue culture.

Much of the success on modern agriculture dependson the availability of high quality seeds with good genetic potential and proven performance in germination, emergence, and vigorous Vegetative growth.

Seed: Seed is a fertilized mature ovule containing an embryonic axis (embryo), stored food material (endosperm) and a protective covering (seed coat or testa).

Seed structures: Living embryo, the most important part of seed, consists of two structures (i) embryonic axis and ( ii ) cotyledon(s). Theembryonic axis is composed of three parts namely

(i)radical (embryonic root), the hypocotyls (point of attachment of cotyledons) and (iii) plumule (theshootapex with the first true leaves). Thethree parts of embryonic axis are easy to identify in dicots, but in monocots (especially in the family Gramineae) their identification is difficult. In monocots, there is only one cotyledon, which is reduced and modifiedto form the scutellum. The basalsheath of cotyledon is elongated to take the shape of coleoptile,while in somecases (e.g., maize), the hypocotyls is modified to form mesocotyl. The base of hypocotyls sheathing the radical is termed as coleorhiza.

Seed development: Thereare manyvariations in the pattern of seeddevelopment when the entire plant kingdom is considered. However, the general processof This includes seed development is almost similar.

Growth and differentiation are regulated by a group of organic compounds known as growth regulators. These growth regulators regulate growth not only at the organ level but also in the plant as a whole. They can be broadly defined as organic compounds synthesized in one part of a plant transported to another where at very low concentration they cause a physiological response.

There are five major groups of growth regulators, namely, the Auxins, Gibberellins, Cytokinins, Ethylene andAbscisic acid. Endogenously synthesized growth regulators are often referred to as plant growth hormones. However, a number of synthetic compounds eliciting similar response as that of the growth hormones have been identified and extensively used. These synthetic hormones effectively substitute the endogenous hormones to understand the basic mechanisms of hormones and also for commercial use.

Photosynthesis is an important physiological process associated with plant growth and productivity. During the process of photosynthesis, the solar energy is trapped and converted into chemical potential which is later used for the reduction of CO₂ into carbohydrates. The powerful reductant, NADPH that is needed for carbon reduction is generated by the light driven reactions of the chloroplast lamellar system. Photosynthesis can be summarized as follows.

During photochemical reactions of photosynthesis, takings place in the chloroplast lamellas system, electrons flow from water to NADP resulting in generation of a powerful reductant NADPH. Simultaneously, water is oxidised and O₂ oxygen is released. This reaction is known as Hills reaction. The rate of production of NADPH and ATP in the photochemical reaction is directly associated with the oxidation of water the sole electron donor. So the photochemical efficiency can be measured by measuring the rate of oxygen evolution.

Since oxygen evolution is a good reflection of the photochemical activity of chloroplast, determination of the rate of oxygen evolution can be conveniently adopted to estimate the photochemical activities. Normally oxygen evolution as a measure of photochemical reactions, is measured either in isolated chloroplasts or in the excised leaf discs using an oxygen electrode. An oxygen electrode is a special form of chemical electrode cell in which the current generated is proportional to the activity of oxygen present. This equipment is widely used for determining the amount of oxygen liberated during the photochemical reaction either in isolated chloroplasts or in leaf bits. Though oxygen electrode is the most frequently used technique to determine the rate of oxygen evolution, there are a few indirect methods to quantify the amount of oxygen evolved.

In photosynthesising tissues or in isolated chloroplasts, the rate of oxygen liberation can be determined by using appropriate artificial electron acceptors and donors (artificial electron donor and acceptors are compounds which can substitute the physiological electron transport carriers and helps in understanding the electron transport mechanisms and ATP synthesis). Oxygen is a powerful oxidizing agent which means it readily oxidizes another compound and itself gets reduced to water. This property can be effectively used in qualitative and quantitative determination of oxygen evolution in green leaves exposed to light. The oxygen that is liberated during the photochemical reactions can be determined by using an appropriate organic substance which can be readily oxidized by oxygen.

Transpiration is the loss of water through areal parts of the plants particularly leaves in the form of water vapour and 95% of the transpiration is through stomata and the rest from cuticle and the lensi cells.

Transpiration can be measured in several ways. The most commonly used technique as the whole plant level is the gravimetric technique where in the amount of water loss through transpiration is measured of monitoring the amount of water added by measuring the loss in weight. Another method is the direct measurement using equipments specially designed for this purpose, which steadily include porometers. Transpiration is also being measured using the portable photosynthesis systems. In the laboratory transpiration rate can be measured using a very simple technique as follows,

Stomata are pores formed between two specialized epidermal cells, the guard cells, which are found on the surface of aerial parts of higher plants (leaves). They control the gas exchange between the leaf and the ambient air. Their main functions is to facilitate the entry of CO² into the leaf for photosynthesis and loss of water vapour for evaporative cooling of the leaves.

The guard cells are formed by the transverse division of stomatal mother cells that differ morphologically and biochemically from other epidermal cells. The middle lamella disintegrates and the two walls separate leaving an opening between them the stomatal pore. The walls bordering the pore become considerably thickened. They are generally cutinised. This radial thickening of the inner wall plays an important role in stomatal movement. Stomata control 95 per cent or more of CO² and water vapour exchange between the leaf and atmosphere.

Location and occurrence of stomata: Stomata are found in all higher plants down to the evolutionary level of the phylum Bryophyta. Stomata are not found in liverworts. In the leaves of most herbaceous plants the stomata are found in both upper (Adaxial) and the lower (Abaxial) surfaces. Such leaves are known as Amphistomatus. Usually in tree species, stomata are found only on the lower surface of the leaves. Such leaves are called Hypostomatus. Aquatic plants with floating leaves such as water lilies have stomata only on the upper surface and such leaves are called Epistomatus. Most of submerged aquatic plants do not posses stomata and diffusion of gases occurs between water and plant tissues across a thin cuticle. In many cases subsidiary cells are present adjacent to the guard cells. These are specialized epidermal cells which are usually different in size and shape from other epidermal cells. They are usually smaller than the epidermal cells and have more dense cytoplasmic contents and a greater frequency of cell organelles. In grasses and some other species, subsidiary cells are intimately involved in stomatal mechanism.

Diffusion of gases through stomata is governed by the two important parameters associated with the stomata, viz., the stomatal frequency and the extent of opening of the stomata. Stomatal frequency is defined as the number of stomata per unit leaf area (no.mm ^-2 ). Apart from the stomatal frequency, yet another parameter that reflects the stomatal distribution is the stomatal Index. It is the ratio of the number of stomata per unit leaf area to the total number of epidermal cells and is generally expressed in percentage.

Objective: To estimate qualitatively various chlorophyll fractions

Materials Required: Leaf material, acetone, petroleum ether, distilled water, methyl alcohol, potassium hydroxide, diethyl ether, separating funnels, stand beakers and conical flasks.

Principle: In biological systems the molecule absorbing radiant energy in the visible range is referred as a pigment. To the human eye these pigments are coloured: chlorophyll is green, carotenoids are red or yellow, xanthophylls is yellow and phytochrone is blue. These pigments are complex molecules. Chlorophylls are the most important pigments active in photosynthesis. At least nine different types of chlorophylls are known – chlorophyll a, b, c, d, e bacteriochlorophylls a and b and chlorobium chlorophylls 650 and 660. Chl a and chl b are best known pigments. In addition to chlorophyll, carotenoids, xanthophylls etc. Are also present as accessory pigments. The chlorophylls are directly involved in photosynthetic electron transport the other pigments serve as accessory pigments and they also serve the purpose of light harvesting.

There are different organic molecules are soluble in different organic solutions, however, their solubility is differential in different chemicals. Based on the nature the chlorophyll pigments can be separated and identified.

A metabolic cell contains about 75-95% water by weight. It is important for plants for proper growth and development. Any significant reduction in availability will affects metabolic activity and thus growth parameters which determine yield. The uptake of water by the plant (roots) from the soil is an osmotic phenomenon. The water potential represents the free energy per moles of water. It can be reduced by:

1. Decreasing temperature
2. Adding soluble pressure or
3. Decreasing pressure or
4. Providing additional absorptive surface

Basic LabTechniques - This laboratory is intended to:

(1) Provide an introduction to basic techniques used in biology labs

(2) provide some experience with the equipment employed in future labs

2.1 Microscopy

The compound light microscope is a veryimportant tool in the study of biology. Typical compound light microscopes allow specimens to be viewed at 400 to 1000 times their actual size (total magnification is determined by multiplying the objective lens magnification by the ocularlens magnification). Thelight microscopeis made up of a system of lenses aligned in such a way thatallows magnification, as well as improved resolution. The resolution obtainable is also dependent on contrastin the specimen. Contrast can be adjusted with the iris diaphragm, located below the condenser lens,and can also be enhanced by the addition of stains to the specimen.

Scientific drawings of specimens observed under the light microscope are often used to provide a record of what was viewed. Proper scientific drawings should be neatly drawn,with smooth lines, in pencil (they should never be in colour). More than one cellshould be drawn to illustrate the cellularassociation to the observer(e.g.found singly or as part of a tissue). Labels should be added using straight horizontal lines to the right side of the drawing, and care should be taken with singular and plural forms of structural terminology. To demonstrate differences in density of specific structures, use stippling rather than shading. A figure caption thatindicates the specimen viewed, how viewed, and the to right side of the drawing, and care should be taken with singular and plural forms of structural terminology. To demonstratedifferences in densityof specific structures, use stippling rather than shading. Afigure caption thatindicatesthe specimen viewed, how viewed, and the magnification of the drawing is always necessary.

Crop physiology is the study of the ways in which plant physiological processes are integrated to cause whole plant responses in communities. The subject matter of crop physiology includes the ways in which the knowledge of plant physiology is applied for better management of crops.

Genetic potential of a plant and its interaction with environmental factors decides its growth and development by influencing or modifying certain internal processes. Plant physiology studies about these internal processes and their functional aspects. It helps to understand various biological processes of the plants like photosynthesis, respiration, transpiration, translocation, nutrient uptake, plant growth regulation through hormones and such other processes which have profound impact on crop yield. Many aspects of Agriculture can benefit from more intensive research in plant physiology to provide practical solutions in agriculture. Understanding the physiological aspects of seed germination, seedling growth, crop establishment, vegetative development, flowering, fruit and seed setting and crop maturity provides a reasonable scientific base for effective monitoring and beneficial manipulation of these phenomenon’s. Studying this phenomenon with a view to develop better crop management practices forms the subject matter of crop physiology. Later on several research works were carried out to understand the processes like translocation of food materials, their partitioning towards economic yield, storage mechanisms, physiology of flowering, effect of stressful environmental factors on crop growth and development, role of plant growth regulators in increasing the crop productivity etc. All these areas have enriched the knowledge of physiological processes and their role in deciding the crop yield. Crop physiology covers different topics such as seed germination and proper establishment of seedling depends upon various internal and external factors. Knowledge in seed physiology helps in understanding of different physiological and morphological changes that occur during germination. Any deviation in these processes causes seed dormancy. The dormant condition of the seed, bars immediate use of harvested seed for next crop which is important in intensive agriculture. By understanding the causes and effects of this problem, crop physiologists have come up with different methods of breaking the seed dormancy. Nutriophysiology is yet another important area to understand crop physiology. For the healthy growth of a crop, around 16 essential elements are required. Knowledge of nutriophysiology has helped in identification of essential nutrients, ion uptake mechanisms, their deficiency symptoms and corrective measures. It also helps to check the toxicity symptoms of various nutrients.

INTRODUCTION

The Ginkgo tree (Ginkgo biloba) is the only living representative of the order Ginkgoales, a group of gymnosperms of the family Ginkgoaceae. Ginkgo tree can live for as long as 1000 years and has been present for over 250 million years (“Living Fossil”). The name Ginkgo is believed to have come from the Chinese word sankyo or yin-kuo, meaning “hill apricot” or “silver apricot”. The species name “biloba,” meaning two lobes, refers to the unique twolobed, fanlike leaves. Ginkgo has been used as both food and medicine for thousands of years. It is the only tree to have survived atomic blast in Hiroshima (Singh et al., 2008).Seeds of Ginkgo were recorded in the early Chinese records on herbals as a medicine.

G. biloba extract (GBE), is the extracts of Ginkgo leaves, and is available as filmcoated tablets, liquid formulations or injectable in markets of Europe and America.

Standardized extract of the leaves of the G. biloba, labeled EGb761, is one of the popular herbal supplements. GBE has been used in traditional Chinese medicine for centuries for treating circulatory disorders, asthma, tinnitus, vertigo, and cognitive problems. Currently, GBE is among the most commonly taken phytomedicines globally, and is approved in Germany for treatment of cerebral insufficiency (memory loss associated with Alzheimer, vascular or multiinfarct dementia) and conditions such as, tinnitus (ringing in the ears), vertigo and intermittent claudication that results in poor circulation (Mohanta, 2012).

INTRODUCTION

Crataegus species, or hawthorn, are shrubs or small trees that have been widely and globally used for treating cardiac problems (arrhythmia, angina, hypertension and gastrointestinal ailments. Fruits of the plant are also consumed as food and nutritional supplement. Crataegus species has several pharmacological properties attributable primarily to presence of flavonoids and anthocyanins. The major flavonoids reported are hyperoside, vitexin, and glycosylated derivatives of these compounds. Crataegus has been listed as the plant for future in terms of its utility and significance. Though the species can be easily grown, its commercial cultivation is not yet popular. This presentation highlights the medicinal significance of the species for its greater utilization.

INTRODUCTION

In India, medicinal plants have traditionally occupied an important place in the socio-cultural, spiritual and health related aspects of peoples’ life. As much as 34% (ca. 6000) of total plant wealth have some known medicinal value (Ved, 2008). The genus Ephedra is one such important group of medicinal plants, from which the drug Ephedrine and a range of medicinally active compounds such as ephedrine, pseudoephedrine, norephedrine, norpseudoephedrine, methyl ephedrine, methyl pseudoephedrine, alkaloids, phenols, terpenoids are obtained. Owing to its medicinal properties, genus Ephedra is in high demand by pharmaceutical industries. It is currently facing heavy extraction pressure from its restricted habitat in the wild, and utterly lacks effective proper conservation and management efforts. For long-term conservation management of this genus, a better and comprehensive understanding of the species remains a top priority, for which detailed studies on its biology, habitat ecology, and agro-technology are required.

INTRODUCTION

Tribulus terrestris L. (Gokhru) is an important industrial herb, belonging to family Zygophyllaceae which includes 285 species representing around two dozen genera. Members of this family are distributed in tropical and warm climates. Most species are shrubs, but some grow as small trees or herbs. T. terrestris is a mat forming plant that grows extensively in warm and dry areas all over the world. In China, India, Greece and other parts of the world, the species finds wide application in folk medicine, in polyherbal formulations and as food supplement for physical rejuvenation, to enhance muscle strength, sexual potency, etc. Amidst many positive claims, clinical trials are recommended to evaluate its reported toxicity in animals.

INTRODUCTION

Himalayan region is known for the diversity of its cultural heritage and climate. In Indian part, this mountain range includes states of Uttarakhand, Himachal Pradesh and Jammu & Kashmir. The climatic variability of the region,from sub-tropical to extreme dry temperate, offers great attraction to naturalists, all across the globe. The forest formations here are the natural habitats of a large number of flora and fauna. The flora found in these forests varies with the altitudinal gradient and is quite distinct. In the sub-tropical Himalayas, for example,the predominant vegetation is of deciduous type consisting of trees, shrubs, scrubs, climbers and xerophytes. Man-made Chir pine forests can also be witnessed. At middle elevation, the climate is sub-temperate and supports a mix of broad leaves and pines forests.In the high reaches including Trans Himalayas, climate is typically wet or dry temperate. In the wet temperate climate, evergreen forests are the predominant vegetation, whereas, the dry temperate areas do not support any significant vegetation.This variation in climate and altitude has a great impact on the biodiversity found in the region, and represents about seven percent of the country’s total biodiversity.

Among this diversity, many valuable and rare medicinal and aromatic plants grow wild in the region. The herbs which are traded from this region are of immense value for the preparation of Ayurvedic drugs in the form of monoherbal preparations or as multi-compound herbal formulations. Charak Samhita mentions that the herbs of the Himalayan forests excel in efficacy.

Abstract

Dactylorhiza hatagirea (D.Don) Soo, a plant species native to Indian Himalayas (Badola and Aitken, 2003; Ved et al., 2003), is one of 110 orchids known for their medicinal properties (Jain 1991; Lal et al., 2004). Known earlier as Orchis latifolia auct. non Linn., D. hatagirea has traditionally been used in indigenous system of medicine in India as ‘Hatajari’ or ‘Salam Panja’. The plant has therapeutic, ornamental and ecological significance (Gupta et al., 1998; Vij et al., 1992; Kala, 2000a). Due to over extraction from nature for medicinal use, the species has become threatened and categorized as critically endangered species (Ved and Tandon, 1998; Kala, 2000b; Ved et al., 2003; Bhattarai et al., 2014). This contribution provides insight into the botanical background, distribution, medicinal importance, threat status, cultivation and commercial aspects, and strategies for its conservation and sustainable utilization.

Introduction

Picrorhiza kurroa Royle ex Benth is an important alpine medicinal plant of the Himalayan region. The root and rhizome of P. kurroa are source of the crude drug, which is characteristically bitter in taste and is linked to the medicinal properties. In Indian system of medicine, the species popularly known as Kutki, is an important constituent of several drugs items. This plant also finds a place in Chinese system of medicine where it is known as Hun-hung lien. Chemically, compounds of interest in Picrorhiza are iridoid glycosides: picroside-I and picroside-II. Material for crude drug has traditionally been extracted from the wild. Natural stands have sharply dwindled due to over extraction and habitat degradation. The species was listed as “endangered” in 1997 by International Union for Conservation of Nature (IUCN). Commercial cultivation is not popular, but is urgently recommended.

Introduction

Species of Polygonatum, traditional known for their therapeutic properties for treating different ailments,are fairly well represented in temperate Northern Hemisphere in Asia, America and Europe and find use. In India, P. cirrhifolium, and P. verticillatum are the two important constituting species of Ashtavarga, but have attained a threatened status due to overexploitation. Of the 60 odd known species of the genus, less than 10 have been explored phyto-chemically. Their status varies species wise, at regional and global level.

INTRODUCTION

The term ‘ginseng’ is applied to a range of plants from family Araliaceae involving genus Panax. Out of the many species available in nature, Himalayas are also home to a commercially lesser known variety called as Panax psuedoginseng Wall (Nayar and Shastry 1990). In the interior temperate areas of the eastern Himalayas, the species is distributed across Sikkim, Bhutan, Nepal, and extends into China. Keeping in view its high commercial international demand, CSIR-Institute of Himalayan Bioresource Technology (CSIR-IHBT) Palampur has recently taken an initiative to introduce and cultivate Chinese ginseng (Panax ginseng) in the Lahaul valley of Himachal Pradesh, in India. This chapter addresses the different biological, chemical, medicinal and cultivation aspects of both the indigenous Himalayan variety (P. psuedoginseng) and the introduced Chinese species (P. ginseng) in particular.

An overwhelming proportion of plant material used for compounding herbal medicines and nutraceuticals is currently collected from the wild. With traditional health care systems gaining increasing popularity, the enormous amount of plant material extracted from the limited plant populations available in natural stands is a cause of justifiable alarm. The unscrupulous plunder of plant populations growing in the wild is pushing increasing number of important species to the endangered category and many to the verge of extinction. Also, unethical practices like adulteration and storage of medicinal plants under unacceptable conditions are rampant. All this is happening, when the consumer is becoming increasingly quality conscious and is demanding traceability of source material used for formulations and adherence to good practices for the entire chain of manufacturing process for herbal drugs and plant-based nutraceuticals. Clearly, solution to this problem lies in replacing the ‘collection from the wild’ paradigm to ‘production by cultivation’. An essential prerequisite for meeting this objective is a well thought out research agenda which not only tackles productivity enhancement goals for the targeted plant species but also addresses the validity of the entrenched notion among healers who use the traditional medical systems, that only material collected from the wild produces the desired results. The reason advanced in support of this argument is that phytomedicines are derived from plant populations growing in the wild under organic conditions and under their natural rhizosphere and rhizoplane. The associated soil-microbial complexes, and a variety of endophytic microorganisms, may be the initiators or modifiers of the production and expression of medicinal properties.

Building and honing up skills for managing any business is an important endeavor. Regardless of whether one is running a small business or a large company individual with good management skills are always essential. The role and structure of the farm business in modern agriculture is changing.

12.1 Supply Chain Management

Supply chain management encompasses the planning and management of all activities involved in sourcing and procurement, conversion and all logistics management activities. Importantly, it also includes coordination and collaboration with channel partners, which can be suppliers, intermediaries, third party service providers, and customers. In essence, supply chain management integrates supply and demand management within and across companies.

11.1 Time Management

One of the greatest assets business can have is an employee who uses his or her time to its fullest potential. Time is something that we can lose and never get back. People are always wishing they had more hours available in the day. The solution to this common problem is easier than one might realize. With a few minor adjustments in how to utilize time, people can change how to manage time. Proper time management can change life on a fundamental basis for the better.

Time management is not an instinct and instead, must be learned. . Using time wisely allows a person to complete tasks in an allotted time period, take on additional work, or spend more time with family. There are many things in an office environment that prevent effective time use.

Interruptions are time-wasters, especially since items one may consider urgent are not necessarily important. There is no way to pre plan the time and duration of an interruption, which may cause a loss of train-of-thought, difficulty concentrating, or complete memory lapse. Although it is difficult to avoid interruptions, there are methods for limiting their duration and impact the schedule.

Time is not like other resources which can be stored and used in the future, Time is a very special resource that you cannot be stored or saved for later use. Everyone has the exact same amount of time each day. Time not well used cannot be retrieved. Most people, feel like they have too much to do and not enough time. They blame lack of time for their poor finances, stress, bad relationships, and for not exercising their body.

Wise time management can help people find the time for what one desire, and for what they need to do. People need time to get what they want in life. If they wait for extra time to appear, they might lose the game of life. Through right time management, they can “create” the time they need, and not just wait for it to come. By planning time wisely, they will have more time to do more things.

10.1 Leading Self and Other

Leading up

In order to develop the ability to be a good Leader, people must learn to lead up with their leader, lead across with colleagues, and lead down with followers. Each of these skills draws on different principles. Leading up is the process of influencing a leader. The principles of leading up greatly increase the chance for success.