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Era of nanotechnology has played a significant role in various aspects of our life. These are minute particles having vast roles. Numerous techniques have been employed for its synthesis. Previously chemical and physical approaches were exploited for their synthesis but nowadays researchers are leaning on biological entities for their creation. And this terminology is green chemistry which does not harm the environment. Mycogenesis plays a terrific role in producing nanoparticles as they contain various enzymes and proteins playing a role in reducing the metal nanoparticles. Metal nanoparticles such as silver nanoparticles act as an efficient biocontrol agent. They were explored to control different types of pests, pathogens, microbes, etc. Different mechanisms were used for controlling the pathogens. They are effective due to broad-spectrum efficiency and ruin the cells by binding with phosphorus and sulphur present in the structure of protein and DNA. They are toxic to a wide range of microorganisms. This chapter focuses on the synthesis of silver nanoparticles using different fungal agents and the processes involved. Further, how the prepared particles have prospective application in the control of living organisms, description of all the pathogens against which silver nanoparticles were effective has been provided. This review will comprehensively provide some knowledge regarding the biocontrol application of myconanotechnology.

Biorefinery produces a spectrum of marketable products (food, feed, materials and chemicals) and energy (fuels, power and/or heat). The term “biorefinery” appeared in 1990 in response to various industry trends. Four conversion-processing techniques depending on the feedstock and the desired output are applied in a biorefinery. These techniques include biochemical (e.g., fermentation and enzymatic conversion), thermochemical, chemical and mechanical processes (e.g., fractionation, pressing and size reduction). The use of these nanoparticles as heterogeneous catalysts in the biorefinery has a great potential to produce biofuel. Although microalgal biorefineries have great potential in the field of biofuel production, certain technical and economic challenges still needed to be addressed. The conversion of extracted microalgal lipid, oils to the fatty acid methyl ester or diesel-range alkanes is the final step in biodiesel production in a microalgal biorefinery. A combination of biorefineries processes with the nanotechnologies is a significant technical advancement in the economics of biorefineries in terms of yield and product range.

The chitosan nanotechnology has been applied for immobilization of enzymes and in turn for biofuel production. This chapter discusses various chitosan nanomaterials and their production technologies as well as their role in biofuel production. Nanoparticles can enhance the strength of chitosan. Their small size and impact on quantum make them better alternatives for the preparation of nanomaterials. The major application of nanomaterials is to provide stability and enhanced activity of catalyst along with better reusability. Chitosan nanoparticles can be fabricated using cross-linking methods. Chitosan-coated magnetic nanoparticles were cross-linked to xylanase and cellulase and were characterized by a x-ray diffraction method. The major limitation of using enzymes for biofuel production is the high cost involved. The biofuel production process has been affected by shear stress that has, in turn, created a negative impact on immobilized lipases. The biofuel industry has employed the application of nanotechnology and nanomaterial for developing cost-effective and process efficient technology to improve biofuel production.

Wide sustainability and reusability of biomacromolecules such as carbohydrates and proteins-based biopolymers pave the way for providing maximal importance in the field of generating biopolymeric nanoparticles. As compared to synthetic nanomaterials, carbohydrate and protein based biopolymeric nanomaterials offer unique advantages that include antibacterial, biocompatible, immunogenicity, and biodegradable properties. Additionally, they have the significant property of more size distribution. Carbohydrate nanoparticles are primarily derived from the polysaccharide biopolymers such as alginate and chitosan; and protein nanoparticles are made from the diverse peptide biopolymers such as albumin, keratin, sericin, fibroin, gelatin and collagen. Advanced methods such as emulsification, desolvation, electrohydrodynamic atomization and coacervation are employed for the controlled fabrication of green biomacromolecules based nanoparticles. Suitability of biopolymeric nanoparticles in plethora of biotechnological applications are quite feasible with the advent of advanced technologies such as dynamic light scattering, scanning electron microscopy, transmission electron microscopy, atomic force microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and UV visible spectroscopy etc. Applications of such biomacromolecules nanoparticles are highly prevalent in agriculture, food, and biomedical industries. Thus, contributions of biopolymeric nanoparticles derived from carbohydrates and proteins biomacromolecules and their recent trends of patents granted in the biotechnological applications are critically discussed along with a promising future scope.

Modern consumers ' interest in natural compounds instead of synthetic bioactive compounds leads to the use of bioactive compounds from natural sources. Microalgae are one of the most important source of many novel nutraceutical and pharmaceutical compounds. About 15,000 algal novel compounds have been identified so far. These bioactive compounds belong to carotenoids, fatty acids, enzymes, peptides, toxins, and sterols, etc. Major algal carotenoids include astaxanthin, β-cryptoxanthin, canthaxanthin lutein and zeaxanthin. Their production depends upon many variables including, nutrient supplements, light, temperature, pH and so forth. Researchers are ceaselessly institutionalizing the algal cultivation conditions to expand the yield and productivity of these high esteemed mixes so that these can be available at a lower cost to the shoppers. In this chapter, an attempt has been made to summarize the various studies conducted to enhance the yield and productivity of two high valued important algal compounds including astaxanthin and lutein under various growth conditions with their algal sources and chemistry.

Flavonoids play a vital role in the biotechnology industry due to many health benefits. The health benefits have been conferred due to unique properties of flavonoids bio-actives. Such unique properties play a vital role in the free radical scavenging, anti-tumour, anti-atherosclerotic, and anti-diabetic etc. Flavonoids are the diverse bioactive compounds that can be classified into various classes such as flavone, flavonol, flavanol, flavanone, isoflavone, anthocyanidin, and chalcones. Flavonoids can be extracted from a diverse range of sources ranging from microorganisms to higher plants. However, extraction of flavonoids from the native source is a tedious process. Hence, various strategies for the flavonoid extraction has been employed. This chapter discusses the physical and chemical methods employed for flavonoid extraction. Various applications of the flavonoids in the biotechnology sector has been elaborately discussed.

Food processing is a series of mechanical or chemical operations in which raw foodstuffs are made suitable for consumption, cooking, or storage. In addition to adding value to the food, it also produces a lot of waste product. According to the Food and Agriculture Organization, one-third of the food is wasted during processing. The food waste produced during processing also leads to various social, economic and environmental problems. With the growing population and increase in demand for processed food, food waste is becoming a critical challenge in the food processing industry. Food wastes are full of many bioactive compounds such as carbohydrates (starch, cellulose, and hemicelluloses), lignin, proteins, lipids, organic acids, smaller inorganic parts minerals, phenolic compounds antioxidants, and vitamins. These potential bioactive compounds can be used in the food, pharmaceutical, cosmetics, and textile industries. Therefore, in the present chapter, an attempt has been made to summarize the various studies conducted to recover the bioactive compounds from food wastes and their utilization as nutraceuticals as well as pharma products. In many countries, the food waste can be processed using a process called valorization which is essentially the process of converting waste materials into more useful products including chemicals, materials, and fuels such as conversion of orange peel into vanillic acid which has diverse application in food and cosmetic industry. Overall, the recovery and utilization of these compounds from food wastes will lead to achieving the goal of sustainability in food production and consumption.

Natural food colorants with functional properties are of increasing interest. Prior papers indicate the chemical suitability of sorghum leaf 3-deoxyanthocyanidins as natural food colorants. Via mutagenesis-assisted breeding, a sorghum variety that greatly overaccumulates 3-deoxyanthocyanidins of leaf tissue, named REDforGREEN (RG), has been isolated and characterized. Interestingly, RG not only caused increased 3-deoxyanthocyanidins but also caused increased tannins, chlorogenic acid, and total phenolics in the leaf tissue. Chemical composition of pigments was established through high-performance liquid chromatography (HPLC) that identified luteolinidin (LUT) and apigeninidin (APG) as the main 3-deoxyanthocianidin species. Specifically, 3-deoxyanthocianidin levels were 1768 μ gg �1 LUT and 421 μ gg �1 APG in RG leaves compared with trace amounts in wild type, representing 1000-fold greater levels in the mutant leaves. Thus, RG represents a useful sorghum mutagenesis In Sorghum bicolor (L.) Moench (sorghum), the most common anthocyanidin types are the 3-deoxyanthocyanidins, primarily luteolinidin (LUT) and apigeninidin (AP), and their glycosylated derivatives, 7 known as 3-deoxyanthocyanins. The 3-deoxyanthocyanins are rare in nature and are of increased interest as natural food colorants due to their better stability to food-processing conditions than the more common anthocya- nins found in most food plants. 7,8 Despite their potential as natural food ingredients, commercially viable sources of the 3- deoxyanthocyanins have not been identified. High concen- trations of the 3-deoxyanthocyanins are reported in black sorghum bran; 9 however, the bran tissue is a limited component of the total plant biomass. In addition, the 3- deoxyanthocyanin pigments are relatively difficult to extract from bran tissue due to their strong association with cell wall material. Recent findings demonstrate that the more abundant plant tissues, leaves and sheaths, of sorghum may be more

This review highlights discoveries made using phage display that impact the use of agricultural products. The contribution phage display made to our fundamental understanding of how various protective molecules serve to safeguard plants and seeds from herbivores and microbes is discussed. The utility of phage display for directed evolution of enzymes with enhanced capacities to degrade the complex polymers of the cell wall into molecules useful for biofuel production is surveyed. Food allergies are often directed against components of seeds; this review emphasizes how phage display has been employed to determine the seed component(s) contributing most to the allergenic reaction and how it has played a central role in novel approaches to mitigate patient response. Finally, an overview of the use of phage display in identifying the mature seed proteome protection and repair mechanisms is provided. The identification of specific classes of proteins preferentially bound by such protection and repair proteins leads to hypotheses concerning the importance of safeguarding the translational apparatus from damage during seed quiescence and environmental perturbations during germination. These examples, it is hoped, will spur the use of phage display in future plant science examining protein-ligand interactions.

This study reports an appreciable yield of podophyllotoxin (PDT) in P. hexandrum plants grown ex situ under polyhouse conditions of a temperate locale. The PDT content of below-ground parts was affected by both plant age and growth period. However, only the effect of plant age on PDT content was significant. Thus, the highest amounts of PDT were recorded in the below-ground parts of 2-year-old plants harvested during the late-growth period (LGP). High total soluble sugars in the below-ground parts during the early growth period (EGP) and the highest nitrate and nitrate reductase in the leaves of 2-year-old plants during the peak-growth period (PGP) indicated higher mobilization and assimilation of starch and nitrate. Probably the surplus carbon and nitrogen gained during the PGP were diverted from aerial parts to below-ground parts during the LGP and in turn contributed to the synthesis of higher amounts of PDT. This study shows that commercial cultivation of P. hexandrum is possible under ex situ temperate conditions.

In silico analysis of amino acid sequences of some aromatic and aliphatic microbial nitrilases for certain physiochemical properties and specificity to aromatic or aliphatic nitriles has been done. The multiple sequence alignment analysis of amino acid sequences has shown clear differences between aromatic and aliphatic nitrilases in terms of position specific presence of conserved amino acid. Statistical analysis of most of the physiochemical parameters did not show any clear distinction between the two nitrilases. In aromatic nitrilases the conserved amino acid residues besides active site domain triad (Glu, Lys, Cys) were His-129, Asn-168 and Arg-174 and these were replaced by Arg-129, His -168 and Lys-174 in aliphatic nitrilases. The physiochemical properties of these two groups of nitrilases also differed e.g. as compared to aliphatic nitrilases, aromatic nitrilases have lesser number of amino acid residues, lower molecular mass, higher pI values, higher content of Ala and Cys residues.

As an adaptive mechanism, hypocotyl dormancy delays emergence of functional leaf until favorable season of growth in Podophyllum hexandrum, an endangered medicinal plant of the western Himalayas. However, upon exposure of the freshly germinated seedlings to favorable temperature (25 degrees C), functional leaves emerged within 20 days. Therefore, we examined regulation mechanisms of growth and development of this alpine plant by temperature under laboratory conditions. The seedlings were exposed to (1) 25 degrees C (temperature prevailing at the time of maximum vegetative growth), (2) 4 degrees C (mean temperature at the onset of winter in its natural habitat), and (3) 10 degrees C (an intermediate temperature). Slackened growth at 4 degrees C was followed by senescence of aerial parts and quiescence of roots and predetermined leaf primordia. Rapid development of leaf primordia at 25 degrees C was associated with increased starch hydrolysis. This was evident from higher alpha-amylase activity and reducing sugars. These parameters decreased on sudden exposure to 4 degrees C. In contrast, the roots (perennating organs) showed a slight increase (1.36-fold) in alpha-amylase activity. Growth and development in seedlings growing at 10 degrees C (temperature less adverse than 4 degrees C) were comparatively faster. The content of reducing sugars and alpha-amylase activity were also higher in all the seedling parts at 10 degrees C as compared to 4 degrees C. This indicated larger requirements for sugar by the seedlings at 10 degrees C. Irrespective of temperature, maximum changes in nitrate and nitrate reductase occurred during the initial 10 days, i.e., when the readily available form of sugars (reducing sugar) was highest. This indicated that a temperature-dependent availability of carbon, but not temperature itself, was an important regulator of uptake and reduction of nitrogen.

This study shows an effective but simple method of conserving characterized populations and elite clones through vegetative propagation and genetic diversity through seeds in Podophyllum hexandrum (family Berberidaceae). Seed dormancy has been considered to be a major constraint in these seeds and most of the earlier reports recommended dormancy-breaking pretreatments such as chilling, gibberellic acid (GA(3)), etc. However, seeds of the 14 accessions that we tested exhibited no dormancy and hence did not require any pretreatments. Besides accession, collection of seeds with high moisture content could be one of the reasons for lack of dormancy. Thus, we propose germination of seeds (while they still retained moisture) in sand at 25 degrees C for high and reproducible results within a shorter period of time compared with earlier reports. Hypocotyl dormancy is known to considerably delay plant establishment and hence en masse propagation by preventing the emergence of functional leaves for up to 11-12 months. Manual removal of cotyledonary leaves, being labor and time intensive, is not a feasible method for large-scale seedling establishment. However, in this study, we showed that GA(3) at 200 ppm can alleviate hypocotyl dormancy besides reducing the time taken for true or functional leaf emergence. Treatment of cotyledonary leaves of 1 week-old-seedlings with 200 ppm GA(3 )resulted in true or functional leaf emergence within 7 days, and the resultant plants were also more vigorous than the ones obtained from manual removal of cotyledonary leaves. The study helped advance the establishment of seedlings by one growing season (almost 1 year).

The endangered Himalayan plant Podophyllum hexandrum syn. P. emodi is inherently slow growing. The plants exhibit delayed emergence of functional leaves or hypocotyl dormancy. However, on GA3 treatment the functional leaves were found to emerge at a favorable temperature of 25°C in a higher percentage of seedlings and in a shorter time. Functional leaves emerged even at 10°C, a temperature when hypocotyl dormancy generally prevails. A considerable increase in the biochemical parameters related to carbon and nitrogen metabolism [starch, sugars and soluble nitrates, α-amylase and nitrate reductase (NR) activity], respiration and total dehydrogenase activity in all the seedling parts also indicated an enhancement of metabolic processes as influenced by GA3, for further growth and development. Specific leaf area of the green cotyledonary leaves increased at 25°C, probably to meet the carbon and nitrogen requirements for new structure formation. Higher activity of enzymes involved in carbon and nitrogen metabolism, i.e., NR and α-amylase especially at Hbn, the region of leaf meristematic activity, was further indicative of higher metabolism for earlier initiation of rapid vegetative growth. Initiation of reserve accumulation was also observed at 25°C.

CELLULOSE SYNTHASE5 (CESA5) synthesizes cellulose necessary for seed mucilage adherence to seed coat epidermal cells of Arabidopsis (Arabidopsis thaliana). The involvement of additional CESA proteins in this process and details concerning the manner in which cellulose is deposited in the mucilage pocket are unknown. Here, we show that both CESA3 and CESA10 are highly expressed in this cell type at the time of mucilage synthesis and localize to the plasma membrane adjacent to the mucilage pocket. The isoxaben resistant1-1 and isoxaben resistant1-2 mutants affecting CESA3 show defects consistent with altered mucilage cellulose biosynthesis. CESA3 can interact with CESA5 in vitro, and green fluorescent protein-tagged CESA5, CESA3, and CESA10 proteins move in a linear, unidirectional fashion around the cytoplasmic column of the cell, parallel with the surface of the seed, in a pattern similar to that of cortical microtubules. Consistent with this movement, cytological evidence suggests that the mucilage is coiled around the columella and unwinds during mucilage extrusion to form a linear ray. Mutations in CESA5 and CESA3 affect the speed of mucilage extrusion and mucilage adherence. These findings imply that cellulose fibrils are synthesized in an ordered helical array around the columella, providing a distinct structure to the mucilage that is important for both mucilage extrusion and adherence.

The endangered alpine plant Aconitum heterophyllum was conserved under ex situ conditions (greenhouse and open) of Palampur by overcoming seed dormancy and plant establishment through hot water treatment (40–60 °C for 30–120 s) of seeds. High seed germination (88 and 76%) was obtained when seeds were treated with hot water at 45 and 50 °C for 90 s. Correlation studies also revealed that treatment enhanced vegetative growth and reproductive yield of the plants. The total alkaloid content of the roots and rhizomes of the plants growing under ex situ conditions was lower than the ones collected from the natural habitat in the 1-year-old plants. However, in the 2-year-old plants, it was almost at par with the ones collected from nature. The life cycle of the plants growing under ex situ conditions also did not vary largely from the plants growing under in situ conditions. Therefore, the present study indicates the successful adaptation of A. heterophyllum plants in conditions other than their natural habitat and hence its potential for sustainable commercialization.

Relationship of leaf anatomy with photosynthetic acclimation of Valeriana jatamansi was studied under full irradiance [FI, 1 600 μmol(PPFD) m−2 s−1] and net-shade [NS, 650 μmol(PPFD) m−2 s−1]. FI plants had thicker leaves with higher respiration rate (RD), nitrogen content per unit leaf area, chlorophyll a/b ratio, high leaf mass per leaf area unit (LMA), and surface area of mesophyll cell (Smes) and chloroplasts (Sc) facing intercellular space than NS plants. The difference between leaf thickness of FI and NS leaves was about 28 % but difference in photon-saturated rate of photosynthesis per unit leaf area (PNmax) was 50 %. This indicates that PNmax can increase to a larger extent than the leaf thickness with increasing irradiance in V. jatamansi. Anatomical studies showed that the mesophyll cells of FI plants had no open spaces along the mesophyll cell walls (higher Sc), but in NS plants wide open spaces along the mesophyll cell wall (lower Sc) were found. Positive correlation between Sc and PNmax explained the higher PNmax in FI plants. Increase in mesophyll thickness increased the availability of space along the mesophyll cell wall for chloroplasts (increased Sc) and hence PNmax was higher in FI plants. Thus this Himalayan species can acclimate to full sunlight by altering leaf anatomy and therefore may be cultivated in open fields.

Using recombinant phage as a scaffold to present various protein portions encoded by a directionally cloned cDNA library to immobilized bait molecules is an efficient means to discover interactions. The technique has largely been used to discover protein-protein interactions but the bait molecule to be challenged need not be restricted to proteins. The protocol presented here has been optimized to allow a modest number of baits to be screened in replicates to maximize the identification of independent clones presenting the same protein. This permits greater confidence that interacting proteins identified are legitimate interactors of the bait molecule. Monitoring the phage titer after each affinity selection round provides information on how the affinity selection is progressing as well as on the efficacy of negative controls. One means of titering the phage, and how and what to prepare in advance to allow this process to progress as efficiently as possible, is presented. Attributes of amplicons retrieved following isolation of independent plaque are highlighted that can be used to ascertain how well the affinity selection has progressed. Trouble shooting techniques to minimize false positives or to bypass persistently recovered phage are explained. Means of reducing viral contamination flare up are discussed.

A group of intrinsically disordered, hydrophilic proteins—Late Embryogenesis Abundant (LEA) proteins—has been linked to survival in plants and animals in periods of stress, putatively through safeguarding enzymatic function and prevention of aggregation in times of dehydration/heat. Yet despite decades of effort, the molecular-level mechanisms defining this protective function remain unknown. A recent effort to understand LEA functionality began with the unique application of phage display, wherein phage display and biopanning over recombinant Seed Maturation Protein homologs fromArabidopsis thalianaandGlycine maxwere used to retrieve client proteins at two different temperatures, with one intended to represent heat stress. From this previous study, we identified 21 client proteins for which clones were recovered, sometimes repeatedly. Here, we use sequence analysis and homology modeling of the client proteins to ascertain common sequence and structural properties that may contribute to binding affinity with the protective LEA protein. Our methods uncover what appears to be a predilection for protein-nucleic acid interactions among LEA client proteins, which is suggestive of subcellular residence. The results from this initial computational study will guide future efforts to uncover the protein protective mechanisms during heat stress, potentially leading to phage-display-directed evolution of synthetic LEA molecules.

Morpho-anatomical leaf traits and photosynthetic activity of two alpine herbs, Podophyllum hexandrum (shade-tolerant) and Rheum emodi (light-requiring), were studied under field (PAR>2 000 µmol m−2 s−1) and greenhouse (PAR 500 µmol m−2 s−1) conditions. Mesophyll thickness, surface area of mesophyll cells facing intercellular spaces (Smes), surface area of chloroplasts facing intercellular spaces (Sc), intercellular spaces of mesophyll cells (porosity), photon-saturated rate of photosynthesis per unit leaf area (P Nmax), and ribulose-1,5-bisphosphate carboxylase/oxygenase activity decreased in the greenhouse with respect to the field and the decreases were significantly higher in R. emodi than in P. hexandrum. P. hexandrum had lower intercellular CO2 concentration than R. emodi under both irradiances. The differences in acclimation of the two alpine herbs to low irradiance were due to their highly unlikely changes in leaf morphology, anatomy, and P Nmax which indicated that the difference in radiant energy requirement related to leaf acclimation had greater impact under low than high irradiance.

Background Improving saccharification efficiency in bioenergy crop species remains an important challenge. Here, we report the characterization of a Sorghum (Sorghum bicolor L.) mutant, named REDforGREEN (RG), as a bioenergy feedstock. Results It was found that RG displayed increased accumulation of lignin in leaves and depletion in the stems, antithetic to the trend observed in wild type. Consistent with these measurements, the RG leaf tissue displayed reduced saccharification efficiency whereas the stem saccharification efficiency increased relative to wild type. Reduced lignin was linked to improved saccharification in RG stems, but a chemical shift to greater S:G ratios in RG stem lignin was also observed. Similarities in cellulose content and structure by XRD-analysis support the correlation between increased saccharification properties and reduced lignin instead of changes in the cellulose composition and/or structure. Conclusion Antithetic lignin accumulation was observed in the RG mutant leaf-and stem-tissue, which resulted in greater saccharification efficiency in the RG stem and differential thermochemical product yield in high lignin leaves. Thus, the red leaf coloration of the RG mutant represents a potential marker for improved conversion of stem cellulose to fermentable sugars in the C4 grass Sorghum.