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ThesisItem Restricted Development of lactic acid starter culture based fermented fruit and vegetable juices(Punjab Agricultural University, Ludhiana, 2021) Modi, Ritika; Sahota, ParampalThe lactic acid fermented fruit and vegetable beverages have been developed through controlled fermentation using ten allochthonous high lactic acid producing (0.612-1.35%), phenotypically and genotypically characterized, homo-lactic- Lactic Acid Bacterial (LAB) strains as starter culture consortium. Value-added secondary metabolite enriched Turmeric, Amla, Black carrots (Kanji), and Black pearl grapes-based functional beverages were developed with improved nutritive value (15-35%) antioxidants, (21-63%) polyphenols, (20-42%) flavonoids and (24%) carotenoids; microbial quality (1010 LAB CFU/ml) and acceptability (8±0.2) with shelf life of 90 days. The optimized bioprocess parameters using Box-Behnken Design in Response Surface Methodology with 5% (v/v) (107 CFU/ml) active starter culture for Turmeric beverage -[turmeric 2%w/v), lemon juice (5%v/v), ginger juice (1.5%v/v); dilution ratio (1:3 with sterilised water); salt (1%w/v)]; Amla beverage -[amla juice: guava juice: ginger juice (1:1:1.5 %v/v); dilution ratio (1:3); salt (0.6%w/v)]; Kanji beverage -[black carrots juice (100ml); salt and rye (1.5%w/v); dilution ratio (1:3)]; Grapes beverage -[grapes juice (100ml), lemon juice (8% v/v); dilution ratio (1:1.5); salt (1.2%w/v)], pasteurized at 82ºC for 10-15 sec and fermentation at 37ºC for 28 hrs. A generic HACCP plan determining critical control points on the line was recommended as a food safety tool during the preparation of beverages. Unstructured kinetic model so developed depicts maximum LAB growth at 8th hour and highest ∆pH and Vmax on the 28th hour and 8th hour, respectively. The increase in lactic acid production (0.35, 0.55, 0.96, 0.63 %TA), reduction in pH (4.51, 4.96, 3.76, 2.64) with enhanced functional aspects based on total polyphenols (52.30, 45.58, 41.85, 52.29 gallic acid equivalents mg/100ml) and flavonoids (44.20, 31.13, 43.91, 46.96 quercitin equivalents mg/100ml) with significantly stronger scavenging activities for the 2,2diphenyl-1- picrylhydrazyl (DPPH) radical (74.25, 86.36, 86.91, 69.70%) and ferric reducing power (87.9, 94.4, 108.66, 100.3 μM FeSO4 equivalents) was observed for Turmeric, Amla, Kanji and Grapes fermented beverages, respectively. These bio-interventions showed antimicrobial activity against food borne pathogens Staphylococcus aureus MTCC3906, Listeria monocytogenes MTCC657, Klebsiella pneumonia MTCC109, Escherichia coli MTCC443, Aeromonas hydrophila MTCC173, as well arrested the initial phase of MOLT-4 and CaCo2 cancer cells lines, down regulating the expression of proto-oncogenes and up regulating the tumor suppressor gene exhibiting the antitumorigenic effect. Further, hepatoprotective and hypoglycemic effect studies showed administration of functional lactic acid fermented turmeric and amla beverages (10 ml/kg body weight) for 6 weeks, significantly reverse or reduce the physiological, metabolic damage, and histological alterations equivalent to the hepatoprotective drug Liv52 in alcohol-induced liver damaged and hypoglycemic drug glibenclamide in streptozotocin-induced diabetic Wistar rats, respectively. Further, lyophilised freeze-dried turmeric and amla fermented powders were developed which can be consumed as ready-to-use fermented beverages by reconstitution @2% and @3.2% (with sterilized water) with retention of all properties of freshly prepared beverage prepared.ThesisItem Open Access Evalution of Plant Growth Promoting Rhizobacteria As potemtial Biofertilizer For Broccoli ( Brassica Oleracea L.)(Punjab Agricultural University, Ludhiana, 2022) Sharma, Kavia; Jupinder KaurThe present study was carried out with the aim to develop a plant growth-promoting rhizobacterial inoculant for improving the growth and yield of broccoli. A total of 24 bacteria were isolated from rhizosphere of broccoli collected from the fields of Punjab Agricultural University, Ludhiana and village Sangrur, Mansa, Barnala and Patiala. Biochemical characterization of these rhizobacterial isolates showed that most of these were oxidase, urease, indole, MR and catalase positive and were involved in starch hydrolysis and gelatin liquefaction whereas negative for citrate and VP test. These isolates were screened for multifarious plant growth-promoting traits viz. solubilization of phosphate and zinc, production of IAA, siderophore, ammonia, HCN and growth on nitrogen free medium. Out of 24 isolates, 17 isolates showed solubilization of phosphate on modified Pikovskaya agar with solubilization index ranging from 0.5-4.5 cm and 53.63-178.6 μg/ml in liquid medium. Eleven isolates showed solubilization of zinc oxide (solubilization index: 0.4-1.9 cm), five isolates showed solubilization of zinc carbonate (solubilization index: 0.3-1.2 cm) and zinc phosphate (solubilization index: 0.1-2.5 cm). Siderophore production was exhibited by fourteen isolates (0.3-1.4 cm). All the isolates were positive for auxin (without tryptophan: 2.04- 13.90 μg/ml; with tryptophan: 10.05-32.04 μg/ml) and ammonia production as well as ammonia excretion (1.803.65 μg/ml). The luxuriant growth on different nitrogen free medium was observed for all the isolates and higher HCN production was observed in three bacterial isolates. Out of these 24 isolates, two potential bacterial isolates RP4 and RP5 (which were named as CBF 2) were selected on the basis of PGP traits for evaluation of their potential to promote growth and yield of broccoli under pot as well as under field conditions. In green house experiment, these isolates when used synergistically showed a significant improvement in soil enzymatic activities (viz. dehydrogenase: 26.13 µg TPF formed h-1 g-1 of soil, alkaline phosphatase: 14.02 µg PNP formed h-1 g -1 of soil and urease: 370.80 µg urea hydrolysed formed h-1g -1 of soil) and plant growth parameters (plant height: 45.7 cm, root length: 6.3 cm, stem diameter: 3.9 cm, fresh weight of shoot :167.85 g, dry weight of shoot:35.70 g, number of leaves per plant: 26, days taken to visible curd:56 days, head compactness: 1.36 cm, number of heads/plant: 4 heads, mean head weight: 87 g and floret stalk length: 12.8 cm) over the control as well as other bioinoculants. The field evaluation of these bacterial isolates as consortium along with FYM and inorganic fertilizers was carried out using Palam samridhi variety of broccoli. The combined application of bacterial strains (RP4 and RP5) and FYM resulted in significantly higher soil microbial population and enzyme activities. Significantly higher available nitrogen (137.30 kg ha-1), available phosphorus (34.21 kg ha-1) and available potassium (170.15 kg ha-1) were also observed in treatment having FYM and bacterial inoculants (N75FYM+N25 + CBF 2). Significantly higher plant nutrient uptake of nitrogen (98.87 kg ha-1), phosphorus (24.84 kg ha-1), potassium (182.27 kg ha-1) was observed in (N75FYM+N25+ CBF 2) and higher uptake of micronutrients viz. zinc (75.5 mg/kg), iron (172.16 mg/kg) and manganese content (71.11 mg/kg) were found to be significantly higher in treatment (N100FYM+ CBF 2) having synergistic application of organic and bio-fertilizers. The various growth attributes of broccoli viz. plant height: 42.1 cm, root length: 11 cm, stem diameter: 4.9 cm, fresh weight of shoot: 214.20 g, dry weight of shoot:38.18 g, number of leaves per plant: 29, days taken to visible curd:44 days, head compactness: 1.11 cm, head yield: 153.15 q/acre, number of heads/plant: 6 heads, mean head weight: 88 g and floret stalk length: 12.8 cm was observed to be statistically higher in treatment T10 with combined application of FYM and bacterial isolates (N100FYM+ CBF 2). The study has led to the selection of a plant growth-promoting bacterial consortium as a potential bacterial inoculant for improving the growth, yield and nutrient content in broccoli.ThesisItem Restricted Bioprospecting production of fermented type II low fodmap (fermentable oligosaccharides, disaccharides, monosacchrides and polyols) sourdough(Punjab Agricultural University, 2023) Khanna, Rahul; Arora, RichaFermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAPs) in the food are short chain carbohydrates which are poorly absorbed and easily fermented in the large intestine by colon micro-organisms, thereby, producing acid and gas. These FODMAPs adversely affect human health leading to irritable bowel syndrome (IBS). In this context, sourdough technology can be exploited to reduce the FODMAP levels in food and alleviate the symptoms of IBS. In the present study, three yeast cultures, viz. Saccharomyces cerevisiae, Clavispora lusitaniae and isolate YB1 were compared for dough raising capacity, where yeast isolate YB1 was screened for maximum dough raising capacity (9.20 %). The screened isolate was characterised as Meyerozyma guillermondii by 18SrRNA gene sequencing and analysed by BLAST. Furthermore, the fermentation parameters for maximum dough raising capacity were optimised as inoculum size 13.6 % (2.4 x cfu/ml) and fermentation time 25.9 h using response surface methodology. After optimisation, fermentation of all- purpose flour using YB1 isolate resulted in reduction of total sugars (84.1 %), reducing sugars (79.8 %), glucose (60.2 %), fructose (88.8 %), lactose (39.3 %), sucrose (98 %) and raffinose (30.5 %). Apart, from sugar reduction, the fermented sourdough was found to have enhanced total antioxidants by 42.3 % and decrease in concentration of anti- nutritional compound i.e. phytic acid (16 %) after 25 h. Colour change by the fermentation process was found to be non- significant. Rheological characteristics of the sourdough, viscosity and shear strain decreased by 60 and 50 % respectively, thereby, increasing the porosity of the product. A total of 34 volatile compounds were detected using GC-MS with major contribution of butanoic acid, 2,3- butanediol tetracontane, n-hexadecanoic acid, octadecanoic acid which enhances aroma and shelf life of the fermented food. The present study demonstrates the potential of yeast isolate YB1 for production of low FODMAP sourdough fermented products using all-purpose flour as a promising alternative food for IBS patients.ThesisItem Embargo Evaluation of microalgal-bacterial consortia for waste water treatment(Punjab Agricultural University, 2023) Poonam; Sharma, Shiwani GuleriaThis study investigated the bioremediation abilities of microalgae and bacteria for the treatment of domestic waste water. Two microalgal viz. PL1 and PL2 and eight bacterial strains viz. B1(L1), B2(L1), B3(L1), B4(L1), B1(L2) B2(L2), B3(L2) and B4(L2) were isolated form indigenous population i.e L1 & L2. Both the microalgae isolates had a long exponential phase and continued to remain in this phase till the end of experimental growth period i.e. 30 days. Log phase of all bacterial isolates were observed at 60 hours of incubation except B1(L1) (48 hours). After screening, two microalgae i.e. PL1 & PL2 and four bacterial isolates were selected for efficient wastewater treatment. B1(L1), B3(L1), B2(L2) and B4(L2) were observed to be efficient in wastewater treatment than other bacteria under study. Coexistence of each strain with other was seen without any trace of growth inhibition which represented their compatibility. From the total of 29 tested consortia, most potential consortium was selected based on the treatment efficacy after 15 days of treatment. Developed consortium no. 29 i.e. PL1+PL2+B4(L2) was selected for domestic wastewater treatment as it showed significant percent reduction among all combinations i.e. BOD (82.9%), COD (77.77%), EC (53.8%), TDS (74.3%),Total nitrogen (71.05%) and Total Phosphorus (69.25%). Higher percent reduction in BOD (88.90%), COD (82.35%) was recorded with selected consortia 29 when treatment provided with source of aeration in 30 days of treatment. SEM observations revealed that several bacterial cells were attached to microalgal cells over 10 days of incubation and it led to the conclusion that microalgae PL1 & PL2 and bacteria B4(L2) can live together and efficiently treat wastewater. Molecular identification of members of consortia 29 was performed. It was revealed after 18S rRNA sequencing PL1 was identified as Chlorella variabilis, PL2 was tentatively identified as Desmodesmus and after 16S rRNA sequencing B4(L2) was identified as Paracoccus sp.. Sequence of PL1 and B4(L2) was submitted to NCBI GenBank with accession no. OR122520.1 and OR467496, respectively. Thus, present study concluded that microalgae and bacteria have more potential to treat wastewater when they were applied in form of consortia. The selected developed consortia no. 29 showed highest percent reduction in all physiochemical parameters with source of aeration in 30 days of wastewater treatment. Nevertheless, the assessment of polluted water composition should be completed prior to the designing and application of microalgae or cyanobacteria based remediation technology, since the pollutant removal efficiency is found to be essentially species dependent.ThesisItem Restricted Effect of nano-scale boron fertilizer on foliar and rhizosphere soil microflora, tuber yield and quality in potato (Solanum tuberosum L.)(Punjab Agricultural University, Ludhiana, 2022) Dhiman, DikshaThe present work aimed to study the effect of foliar application of the synthesized boron nanoparticles and bulk precursors on soil and foliar microflora and on yield and quality of potato tubers. Three different types of boron nanoparticles were synthesized i.e., calcium borate, boron nitride and boron tri-oxide nanoparticles. On UV- Vis absorption spectroscopy analysis, the synthesized boron nanoparticles exhibited characteristic absorption peaks at 214 nm (Ca B NPs), 200 nm (BN NPs) and 200-205 nm (B2O3 NPs). The FTIR analysis of these nanoparticles depicted the presence of the borate network and boron-oxygen functional groups. The microbial cell toxicity of the synthesized nanoparticles was evaluated through poison food agar and broth supplementation in vitro studies against plant beneficial soil bacteria and no zones of inhibition were observed. Further, on evaluation of the antioxidant properties of the nano-boron particles significant variations were among the three different types of nanoparticles. In broth supplementation study, the viable cell count varied significantly for these different types of nanoparticles. The total protein content also exhibited a significant effect for BN NPs and B2O3 NPs except for Ca B NPs. The acid production as determined by TLC analysis showed enhanced formation of indole acetic acid in Azotobacter (Rf value: 0.65). A field experiment was performed with the CaB NPs to ascertain effect of foliar application on cracking and tuber yield in two potato accessions. The nano boron fertilization exhibited non-significant effect on all the soil parameters. The soil dehydrogenase activity showed significant variations however both the varieties, treatment and different concentrations were at par with each other for the urease activity. A significant effect on the shoot fresh weight (variety specific). The variety and various boron source treatments at different concentrations exhibited a non-significant effect on all the photosynthetic parameters except for the anthocyanin content. Non-significant difference was recorded for the yield and yield attributing parameters such as tuber number, diameter and fresh weight. Further, nonsignificant results were obtained for all the tuber quality parameters. Higher N and B content of the shoot was recorded at 50 mg L-1 and higher N and B content in the tuber was recorded at 100 mg L-1 nano boron. The results reflect the equivalence for bulk and nano-sources of boron with a numerical increase in the observed traits by application of nano-boron particles.ThesisItem Restricted Effect of slow release urea nano fertilizer on soil microflora and yield of direct seeded rice (Oryza sativa L.)(Punjab Agricultural University, 2023) Yashika; Kalia, AnuThe present work aimed to study the effect of a slow release urea nanofertilizer on soil microflora and yield of direct seeded rice (Oryza sativa L.). A soil column study indicated slow-release pattern of the urea from the nano-urea product varied for the soil samples having three different textures (low, medium and heavy).The leachate analysis revealed release of ammonical and nitrate nitrogen from the synthesized product. Further, the effect of the nanourea fertilizer on soil microflora study identified that the viable cell counts for a variety of soil microbes were not negatively affected on application of different doses of nanourea fertilizer. The soil urease, protease, and dehydrogenase enzyme activities exhibited significant increase in nano urea (100% RDN) treatment in light textured soil. A field experiment was performed to observe the effect of nano urea fertilizer on the growth and yield of direct seeded rice. An increase in the plant height, shoot and root fresh weight and dry weight was recorded for different doses of Nitrogen applied through nano urea and conventional urea N-fertilizer level for plants sampled after every 30 days after sowing till three months of the growth. The maximum plant growth was recorded for treatments which received 100% of the recommended N-fertilizer. Likewise, the yield attributing traits particularly the number of filled grains and thousand grain weight increased on application of 100% Recommended dose of N through nano urea. The soil chemical parameters were recorded to be non-significantly altered for plant and grain nutrient content (NPK content) at nano-urea fertilizer (100% RDN). Non-significant effect was recorded for the soil microbial viable cell count on different doses of nano-urea fertilizer. These results indicate the use of nano-urea fertilizer as a possible alternative for conventional urea fertilizer.ThesisItem Restricted Evaluation of mushroom growth promoting microbial isolates for enhancement of yield potential in Calocybe indica(Punjab Agricultural University, 2023) Navneet Kaur; Gagandeep KaurMushroom growth promotion by means of biological agents is an emerging aspect in mushroom cultivation. The present study was aimed to search for a novel bacterium potentially able to enhance the growth and yield of Calocybe indica, also known as milky mushroom. Soil samples from 6 sites supporting natural growth of macro-fungi were collected and screened for isolates. Out of the total 79 isolates, 5 isolates were selected for further analysis on the basis of mushroom growth promoting traits. During the in vitro radial and linear growth rate evaluation through co-cultivation of isolates with C. indica mycelium, isolate B28 was observed to enhance the mycelial growth (7.10±0.10 mm/day) as well as the linear growth rate (4.94±0.14 mm/day) compared to control (6.47 and 4.30, respectively). Isolate B28 was used at various concentrations during the cultivation of C. indica. It has been observed that the number of days of spawn run were reduced in case of supplementation at the rate of 5 ml/bag (20±1). Percentage increase biological efficiency (20.63%) of the crop was also observed in case of 5ml supplementation of isolate in comparison to control. At higher concentrations (25 ml/bag), biological efficiency was found to be decreased by even as much as 86.78%. It can be concluded that the use of isolated bacteria (B28) at an optimal concentration (3ml and 5 ml/bag of 108 CFU/ml culture) in both the substrate and casing layer can decrease the number of days of spawn run, number of days for pinning, and can increase the number of fruiting bodies, and yield of the crop, hence resulting in increase in biological efficiency.ThesisItem Restricted Evaluation of plant growth promoting rhizobacteria to develop biofertilizer for pumpkin (Cucurbita pepo L.)(Punjab Agricultural University, 2023) Manmeet Kaur; Jupinder KaurThe present study was carried out with the aim to assess the potential of plant growth-promoting rhizobacterial inoculant for improving the growth and yield of pumpkin. A total of 56 rhizobacteria were isolated from the rhizospheric regions of pumpkin collected from the fields of Punjab Agricultural University, Ludhiana and village Sangrur, Ahmedgarh, Jalandhar and Muktsar. Primary screening was done on the phosphate solubilizing ability of these isolates. All the 56 isolated rhizobacteria were assessed for their potential to solubilize inorganic phosphorous. Out of these 56 isolates, only 20 isolates showed solubilisation of phosphate on modified Pikovskaya agar with solubilization index ranging from 0.5-4.2 and 50.15 to 180.42 µg/ml in liquid medium. Biochemical characterization of these rhizobacterial isolates showed that most of these were positive for oxidase, catalase, urease, MR, VP, starch hydrolysis, gelatine liquefaction and indole synthesis test whereas negative for citrate utilization test. These isolates were screened for other multifarious plant growth-promoting traits viz. solubilization of zinc; production of IAA, siderophore, ammonia, HCN and growth on nitrogen free medium. Nine isolates showed solubilization of zinc oxide (solubilization index: 0.1-2.5), ten isolates showed solubilization of zinc phosphate (solubilization index: 0.1-3.0) and seven isolates showed solubilization of zinc carbonate (solubilization index: 0.1-1.5). Siderophore production was exhibited by twelve isolates (0.5-2.0 mm). All the isolates were positive for auxin (without tryptophan: 1.45-12.66 μg/ml; with tryptophan: 8.92-32.67 μg/ml) and ammonia production as well as ammonia excretion (1.015 to 7.237 μg/ml). The luxuriant growth on different nitrogen free medium was observed for all the isolates. Out of these 20 isolates, two potential bacterial isolates RA4 and RM2 (which were named as CBF 2) were selected on the basis of PGP traits for evaluation of their potential to promote growth and yield of pumpkin under pot as well as under field conditions. In green house experiment, these isolates when used synergistically showed a significant improvement in soil enzymatic activities (viz. dehydrogenase: 24.4 µg TPF formed h-1 g-1 of soil, alkaline phosphatase: 14.37 µg pNP formed h-1 g-1 of soil and urease: 373.56 µg urea hydrolysed formed h-1g of soil-1) and plant growth parameters (number of leaves per plant: 30, number of flowers: 24, available nitrogen: 218.44 kg/ha, available phosphorous: 51.55 kg/ha and available potassium: 144.38 kg/ha) over the control as well as other bioinoculants. The field evaluation of these bacterial isolates as consortium along with FYM and inorganic fertilizers was carried out using pumpkin as host. The combined application of bacterial strains (RA4 and RM2) and FYM resulted in significantly higher soil microbial population and enzyme activities. Significantly higher available nitrogen (252.27 kg ha-1), available phosphorus (57.10 kg ha-1) and available potassium (147.44 kg ha-1) were observed in treatment having FYM and bacterial inoculants (N75 FYM+N25 + CBF 2). The various growth attributes of pumpkin viz. number of fruits per plant: 5.33, fruit diameter: 14.57 cm, fruit length: 12.43 cm, fruit weight: 2.17 kg, number of seeds per fruit: 322.67, 1000 seed weight: 42.61 g, fruit yield: 33.60 tonn/ha, seed yield: 7.42 q/ha, flesh colour of fruit: yellow, thickness of flesh: 2.20 cm, total sugars: 10.65 g/100 g, zinc uptake by fruit: 0.31 ppm, seed oil yield: 20.01 q/ha and seed oil percentage: 28.27 %, phosphorous uptake by seeds: 41.64 ppm, zinc uptake by seeds: 0.37 ppm was observed to be statistically higher in treatment T10 with combined application of FYM and bacterial isolates (N100FYM+ CBF 2). Based on 16S rRNA gene analysis, the bacterial isolate RA4 was identified as Psuedomonas putida and RM2 as Pseudomonas moraviensis. The study has led to the selection of a plant growth-promoting bacterial consortium as a potential bacterial inoculant for improving the growth, yield and nutrient content in pumpkin.ThesisItem Embargo Characterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.)(Punjab Agricultural University, 2023) Manmeet Kaur; Sharma, ShivaniPleurotus florida and Calocybe indica are the edible mushrooms that have been widely accepted due to their ability to grow on a variety of substrates and possess the potential to produce oxidative and hydrolytic enzymes. The present work involved the study and characterization of oxidative/hydrolytic enzymes of P. florida and C. indica for their potential applications in alcohol fermentation, dye decolorization, biobleaching and mushroom production. The oxidative and hydrolytic enzymes extracted from P. florida and C. indica were estimated at different mycelial growth stages for enzyme production. The intracellular and extracellular enzyme activity of P. florida and C. indica increased with time and maximum activity of lignocellulolytic enzymes was found to be on 14th day of incubation. In both mushrooms, ligninolytic enzyme activity increased during substrate colonisation but quickly decreased during fruiting body development. On the other hand, P. florida and C. indica showed relatively modest hydrolase activity during substrate colonisation. The activity of hydrolytic enzymes increased dramatically during primordial formation and peaked at the fruiting body stage. The purification of laccase, lignin peroxidase, manganese peroxidase and endoxylanase from the fruiting body of P. florida resulted in maximum purification fold of 21.49, 17.73, 16.81 and 12.78 with yield of 24.98%, 20.60%, 19.53% and 14.86%, respectively. However, the purification of laccase, lignin peroxidase, manganese peroxidase and endoxylanase from the fruiting body of C. indica resulted in maximum yield of 21.36%, 18.92%, 17.37% and 16.62% with purification fold of 25.6, 25.12, 20.88 and 19.97, respectively. The SDS-PAGE of the purified enzyme laccase and lignin peroxidase isolated from P. florida showed a single prominent band at 66 kDa and 55 kDa respectively. The SDS-PAGE of the purified enzyme laccase and lignin peroxidase isolated from C. indica showed a single prominent band at 64 kDa and 47 kDa, respectively. FTIR spectra of the purified enzymes indicated a secondary structure that reflected the amide I and amide II bands, respectively. The pretreatment of paddy straw and wheat straw with P. florida, C. indica and their ligninolytic enzyme resulted in decrease in lignin and hemicellulose content, respectively during the incubation period of 30 days. However, the decrease in cellulose content occurred during the pretreatment of substrates with fungus while relative increase in cellulose content during the treatment with ligninolytic enzymes was observed over the incubation period of 30 days. Under optimized conditions, saccharification of biologically pretreated paddy and wheat straw resulted in release of 0.415 and 0.389 g/gds reducing sugars, respectively. The fermentation of biologically pretreated and commercial cellulase saccharified paddy and wheat straw hydrolysate resulted in 0.129 and 0.119 g/g ethanol, respectively. The crude enzyme extract of P. florida and C. indica were able to degrade RBBR (25.74 %, 22.06%) and Amido Black (19.76%, 17.58%) dyes maximally after 96 hours of incubation at 30°C and pH 7.0. The paddy and wheat straw was treated with a ligninolytic crude enzyme, which stimulated faster mushroom growth and fructification. The present study thus revealed that expression of biosynthetic potential of P. florida and C. indica is highly dependent on the method of fungi cultivation. These ligninolytic enzymes showed a unique profile in terms of versatility, greenness, pollutant removal and efficiency in lignin degradation for the exploitation and valorization of agro-wastes.