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Open Access
Bioprocessing of Indian gooseberry for the production of lactic acid beverage and pickles
(Punjab Agricultural University, Ludhiana, 2019) Kirandeep Kaur; Sahota, Param Pal
The plant-based fermented beverages have received considerable interest for their rich nutrition, high content of soluble and insoluble fibers and for their fermentability by lactic acid bacteria (LAB). A set of ten allochthonous lactic acid bacterial strains phenotypically and genotypically characterized for their probiotic potential were used as starter culture for the optimization and production of Indian gooseberry-based functional beverage and lacto-pickle. The optimized bioprocess comprised of gooseberry juice blend [gooseberry juice 2: ginger extract 1: guava juice 2], dilution ratio (1:3), condiment concentration (0.7%), pasteurized at 82ºC for 10 seconds and 5.0% (v/v) (106 CFU/mL) active starter culture for fermentation at 37ºC for 24 hr. The microbiological and physicochemical parameters of stored beverage (90 days) were; 1ºBrix, Titrable acidity 0.30-0.89 %, pH 6.0 -2.6, Total Sugars 44.3-35.3 mg/100mL, Reducing sugars 5.4-10.7 mg/100mL and antioxidant activity (85-67%), Total polyphenolic content (42.5-32.6 mg GAE/100mL),Total flavanoids (14.5-28.7 mg/100mL) with LAB capable of maintaining cell counts of 7.39 log CFU/mL. The bioprocess optimized for the Indian gooseberry lacto pickle as 5.0% (v/v) (106 CFU/mL) active starter culture and 5% brine solution for fermentation at 37ºC for 24 hr. The microbiological and physicochemical parameters of stored lacto-pickle (90 days) were; LAB count-6.22-7.27 log CFU/mL, 1ºBrix, Titrable acidity 0.24-0.64 %, pH 7.0-2.8, Total Sugars 20.1-11.6 mg/100g, Reducing sugars 2.60-14.6 mg/100g, viz. nutraceuticals Total antioxidant activity 86.1- 90.2%, Total polyphenolic content 16.6-37.8 mg GAE/100g and Total flavanoids 4.007-55.7 mg/100g. The overall acceptability of beverage and pickle was 7±0.2 on nine point hedonic scale. The bio-interventions: Gooseberry beverage and pickle endowed with organic acids, metabolites, antioxidants, polyphenols and flavonoids showed antimicrobial activity against food borne pathogens Staphylococcus aureus MTCC3906, Listeria monocytogenes MTCC657, Klebsiella pneumoniae MTCC109, Escherichia coli MTCC443, Aeromonas hydrophila MTCC173 as well as arrested the initial phase of Molt-4 cancer cell lines, down regulating the expression of proto-oncogenes and up regulating the tumor suppressor gene exhibiting the antitumorigenic effect.
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Bioprocessing of Turmeric (Curcuma longa Linn.) for the production of lactic acid beverage and pickles
(Punjab Agricultural University, Ludhiana, 2019) Makkar, Avneet Kaur; Sahota, Param Pal
The plant-based fermented beverages have received considerable interest for their rich nutrition, high content of soluble and insoluble fibers and for their fermentability by lactic acid bacteria (LAB). A set of ten allochthonous lactic acid bacterial strains phenotypically and genotypically characterized for their probiotic potential were used as starter culture for the optimization and production of Turmeric-based functional beverage and lacto-pickle. The optimized bioprocess comprised of turmeric juice blend [Turmeric juice 100mL: ginger extract 1.5mL: lemon juice 5mL], dilution ratio (1:3), condiment concentration (1.0%), pasteurized at 52ºC for 15 seconds and 5.0% (w/v) (106 CFU/mL) active starter culture for fermentation at 37ºC for 24 hrs. The microbiological and physico-chemical parameters of stored beverage (90 days) were; 1ºBrix, titrable acidity 0.30-0.80 %, pH 4.8-2.5, total sugars 48.56-13.0 mg/100mL, reducing sugars 11.54-4.70 mg/100mL and antioxidant activity (70.37-68.58%), total polyphenol content (44.20-40.00 mg GAE/100mL), total flavonoids (38.06-48.11 mg/100mL) with LAB capable of maintaining cell counts of 6.99 log CFU/mL. In addition, the beverage prepared from: 2% turmeric powder (var. Punjab Haldi-1) showed a significant difference in total polyphenol (37.12 mg GAE/100mL) and flavonoid content (43.78 mg/100mL). The bioprocess optimized for the Turmeric-lacto pickle as 5% (w/v) (106 CFU/mL) active starter culture and 5% brine solution for fermentation at 37ºC for 24 hrs. The microbiological and physico-chemical parameters of stored lacto-pickle (90 days) were; LAB count 7.05-5.82 log CFU/mL, 1ºBrix, titrable acidity 0.28-1.12 %, pH 4.6-2.2, total sugars 10.21-7.26 mg/100g, reducing sugars 6.54-7.50 mg/100g, total antioxidants 53.04-72.17%, total polyphenolic content 45.35-57.12 mg GAE/100g and total flavonoids 39.25-45.15 mg/100g. The overall acceptability of beverage and pickle was 8±0.2 on nine point hedonic scale. The bio-interventions; Turmeric beverage and pickle endowed with organic acids, metabolites, antioxidants, polyphenols and flavonoids showed antimicrobial activity against food borne pathogens Staphylococcus aureus MTCC3906, Listeria monocytogenes MTCC657, Klebsiella pneumoniae MTCC109, Escherichia coli MTCC443, Aeromonas hydrophila MTCC173 as well as arrested the initial phase of Molt-4 cancer cell lines, down regulating the expression of proto-oncogenes and up regulating the tumor suppressor gene exhibiting the antitumorigenic effect.
Open Access
Isolation and characterization of Rhizobium spp. for its plant growth promoting potential in Lucerne (Medicago sativa L.)
(Punjab Agricultural University, Ludhiana, 2019) Jotshana Kumari; Khanna, Veena
The present investigation was undertaken to access the effect of rhizobial isolates on plant growth and symbiotic parameters in Lucerne (Medicago sativa L.).Forty rhizobial isolates were identified and characterized culturally, morphologically, biochemically and found belonging to genera Rhizobium. The isolates were functionally characterized on the basis of phosphate solubilization, Indole acetic acid, siderophore, cyanide and gibberellic acid production. Indole acetic acid production ranged from 2.6 -77.7 μg/ml in presence of tryptophan after 3 days of incubation and increased to 2.9-101.7 μg/ml on 5th day.The range of PSI (Phosphate solubilization index) was 13-17 mm with highest solubilizing index shown by LNR-6, LNR-20, LNR-23 and LNR-39 (17mm).Twenty eight of the forty isolates were able to produce siderophore as evidenced on Chrom-azurol-S-agar, maximum catecholate type siderophore produced by LNR-39 (43.4 μg/ml). 57.5% isolates produced HCN out of which 13 % were strong and 39.1% were moderate producers. The gibberellic acid production ranged from 41.2 -96.6 μg/ml with LNR-39 (96.6 μg/ml) being the potent gibberellic acid producer. The study carried out showed that isolates LNR-2,LNR-4, LNR-6, LNR-22, LNR- 32 and LNR-39 exhibited growth and functionality traits which were at par, hence they were selected for field trail. The field experiment revealed enhanced stem and root length,dry weight of nodules, nitrogen content, leghemoglobin and chlorophyll content , green fodder yield under Rhizobium treatment. Thus, the present study revealed that treatment with Rhizobium LNR-39 and treatment LNR-4 were effective for plant growth promotion and enhanced symbiotic effeicency in Lucerne.
Open Access
Biosorption of electronic waste through microbial cultures: a bioremediation technology
(Punjab Agricultural University, Ludhiana, 2019) Preetiman Kaur; Sharma, Shivani
Seven microbial cultures Lentinus edodes, Pleurotus florida, Ganoderma lucidum, Aspergillus niger, Trametes versicolor, Streptomyces spp. and Pseudomonas spp. were evaluated for biosorption and bioleaching from electronic-waste (e-waste) under submerged conditions. Pleurotus florida and Pseudomonas spp. showed the maximum specific activity of laccase enzyme (6.98 U/mg and 5.98 U/mg respectively) on 20th day of incubation in comparison to other microbial cultures. Maximum biosorption and bioleaching of copper was observed in Pleurotus florida (97.26 mg/g and 44.30 mg/l respectively) followed by Pseudomonas spp. (96.20 mg/g and 42.06 mg/l respectively). However, Pseudomonas spp. showed maximum leaching (46.33 mg/l) and absorption (98.00 mg/g) for iron followed by Pleurotus florida (43.13 mg/l and 94.13 mg/g). Presence of large halo orange zones in ewaste treated sample of Pseudomonas spp. confirmed the production of siderophores. The conditions for laccase production using best screened microbial cultures were optimized by Response Surface Methodology using Design Expert 12.0 and regression equations were developed with R2 value ranging between 0.7817 to 0.9257 indicated high degree of fitness. The protein profiling of these optimized cultures by SDS-PAGE revealed the presence of more number of protein bands in e-waste containing samples in comparison to untreated cultures. The topographical studies of samples treated with e-waste through SEM showed the etching of surfaces along with deposition of certain vegetative and spores cells, extracellular material secreted by these microbial cultures and heavy granular pits. Further EDX studies showed the decrease of metal ions from their respective initial concentrations present in untreated samples which confirmed the bioleaching followed by biosorption.
Open Access
Calocybe indica strain evaluation for lignocellulolytic enzymes in relation to mushroom yield
(Punjab Agricultural University, Ludhiana, 2019) Karanpreet Singh; Sodhi, Harpreet Singh
Calocybe indica, milky mushroom is a tropical mushroom of Indian origin that grows well at 28-35°C. C. indica strains were evaluated for their mycelial growth, extracellular enzymes producing capability and their yield potential. Different characteristic of nine strains of Calocybe indica Ci-01 to Ci-09 and a Macrocybe were grown on Potato Dextrose (PDA), Complete Yeast Extract (CYM), Mushroom Minimal Media (MMM) and their broth at 30±2°C. After 10 days, maximum radial growth was observed on PDA in Ci-06 (87.6 mm) and Ci-09 (87 mm) strain of C. indica. The maximum biomass was recorded in CYM in Ci-06 (16.1g/l) and Ci-07 (15.5 g/l). The maximum linear growth on wheat straw after 10 days was observed in Ci-07 (63mm) and Ci-09 (66mm). In wheat straw, the maximum biological efficiency estimated from the harvested yield (kg/q dry straw) was observed in the strain Ci-06 (57.3%). Two strains Ci-07 (48.5%) and Ci-09 (45.7%) were found to give yield at par with each other. Maximum number of fruiting bodies were recorded in the strain Ci-06 (1552 no./q dry straw). In culture filtrate endoglucanase enzyme activity range between 1.91-4.42 U/mg, endoxylanase activity ranged between 2.73-4.76 U/mg, laccase activity ranged between 6.94-10.2 U/mg. During spawn run, endoglucanase and endoxylanase enzyme activity ranged between 0.58-1.03 U/mg and 0.74 -2.11 U/mg, laccase activity was ranged between 1.51-4.54 U/mg. During pinhead, endoglucanase and endoxylanase activity ranged between 0.61-1.59 U/mg and 1.57-2.96 U/mg, whereas laccase activity range between 6.53-10.6 U/mg. In fruiting bodies, endoglucanase and endoxylanase activity of the strains ranged between 0.94-1.88 U/mg and 0.93-2.36 U/mg, laccase activity ranged between 5.43-9.21 U/mg. A positive correlation of yield with biomass and endoglucanase has been observed in C. indica strains Ci-03, Ci-06, Ci-07 and Ci-09.
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Pretreatment of paddy straw for cultivation of Vplvariella (Bull.ex Fr.) Singer
(Punjab Agricultural University, Ludhiana, 2018) Swarthi K.; Sodhi, Harpreet Singh
Two strains of Volvariella volvacea, VV3 and VV6 were evaluated for their mycelia growth rate and yield potential on pretreated paddy straw as substrate under Punjab conditions. Mycelial growth studies on Nutrient agar (NA) and Potato Dextrose agar (PDA) inoculated with Bacillus sp. and Delftia sp. separately revealed that bacteria had no antagonistic effect on the growth of V. volvacea. The mycelia growth rate of V. volvacea on PDA inoculated with bacterium was statistically at par with control and covered plate in 5 days whereas on NA mycelia growth covered the plate in 8 days. The effect of pretreatment of paddy straw on its chemical composition was studied with 6 different pretreatments: urea 1% (w/w), urea 0.5% (w/w), urea 0.5% (w/w)+Delftia sp. 0.5% (v/w), urea 0.5% (w/w) +Bacillus 0.5% (v/w), Delftia sp. 0.5%, Bacillus sp. 0.5% (v/w) and water 0.5% (v/w) as control. The pretreated paddy straw was analysed for proximate principles at a periodic interval of 5 days upto 20 days. Neutral Detergent Fiber (NDF), Acid Detergent Fiber (ADF), Cellulose, hemicelluloses, Acid Detergent Lignin (ADL), Ash content and crude protein content were analysed. A significant decline in NDF value from 75.7% to 46 % and 34.4% was observed when urea 0.5% + Bacillus 0.5% + Delftia 0.5% were used as pretreatment of paddy straw respectively.The ADF value was significantly reduced to 23% and 33% when urea 0.5% +Delftia 0.5% and urea 0.5% + Bacillus 0.5% were used to treat paddy straw. A significant reduction in cellulose content was observed from 40% to 25% to 20% with urea 0.5%+ Delftia 0.5% and urea 0.5% + Bacillus 0.5% respectively. The hemicelluloses content recorded maximum reduction to 11.45% from 23.4% with urea 0.5% + Delftia 0.5%. There was a rapid decline in ADL with urea 0.5% + Delftia 0.5% and urea 0.5% + Bacillus 0.5% from 12.3% to 4.3% and 7% respectively. The ash and crude protein content increased with all the pretreatments. Studies on mycelia growth rate on pretreated paddy straw revealed that the addition of bacteria along with urea had improved the growth rate of both the strains of V. volvacea indicating the significant role of bacterial pretreatment in the mycelia growth of V.volvacea. Cultivation of V.volvacea strains on pretreated paddy straw had shown the maximum yield was obtained from pretreated paddy straw from VV6 (14.84 kg/ q dry straw) followed to strain VV3 (13.82 kg/q dry straw) on pretreated paddy straw. The yield from untreated (control) paddy straw was 10.5 kg/q dry straw from VV3, 12.5 kg/q dry straw from VV6. Present study revealed that pretreatment of paddy straw with urea 0.5%+Delftia sp. 0.5% gave better yield than untreated paddy straw.
Open Access
Cellulose degrading bacteria, Decomposition, Green manure, Maize, Microbial dynamics, Nutrient mineralization, Yield
(Punjab Agricultural University, Ludhiana, 2019) Charnjeet Kaur; Khanna, Veena
Thirty four rhizobacterial isolates were isolated from mungbean rhizosphere soil collected from different locations of the Punjab. The isolates were evaluated for their PGP traits at 35°Cand 40°C but, it was found that rhizobacteria were more acclimatized to 35°C as compared to 40°C. All the isolates showed IAA production at 35°C (0.13- 56.44μg/ml) and (0.22-36.80μg/ml) at 40°C. The isolates were effective P and Zn solubilizers with P-solubilisation index ranging from 6.9-15.5 at 35°C and from 5-13.8 at 40°C. Zn solubilisation index ranging from 3.8-24.0 at 35°C and 4.6-15.3 at 40°C, most potent isolates being G-2, G-3, G-33 and G-34. Isolates G-2, G-3, G-33 and G-34 were HCN and ammonia producers as well. The biocidal impact of the rhizobacterial isolates against F.oxysporum and R.solani was studied and eight isolates showed antagonistic activity against F. oxysporum and R. solani. Isolates G-2, G-3, G-33 and G-34 proved to be strong antagonistics with 27.0-37.5% inhibition of the phytopathogens. The field experiment conducted revealed that co-inoculation of Rhizobium and G-33 proved to be the most beneficial, resulting in the improvement of symbiotic traits viz nodule number per plant (66.6), nodule dry weight (22.2 mg/plant), root (0.29g/plant) and shoot (11.84g/plant) dry weight and the yield (1134 kg/ha) of mung bean (ML2056) as compared to Rhizobium treatment alone (1093 kg/ha).
Open Access
Effect of tillage systems and carbon sequestration on soil micro flora in wheat rhizosphere
(Punjab Agricultural University, Ludhiana, 2019) Tabasum, Shaik; Khanna, Veena
The present work was aimed to evaluate the impact of different tillage systems on soil micro flora and carbon sequestration for two consecutive seasons in rhizosphere of wheat crop. To achieve this objective, rhizospheric soil samples were drawn from different tillage treatments at various crop growth stages from long-term experiment (since 2016) in organically grown basmati rice-wheat cropping system, conducted at Students Research Farm, Department of Agronomy, Punjab Agricultural University, Ludhiana. A total of seven treatments, out of which six were organic and one control i.e., application of recommended dose of agrochemicals were analysed. Organic treatments comprised of different tillage systems, deep tillage (DT), zero tillage (ZT) and conventional tillage (CT) added organic amendment (paddy straw mulch). Different soil physico-chemical parameters soil pH, electrical conductivity, available phosphorous and potassium were significantly increased in conjugation of zero tillage with organic amendments. Microbial and different enzyme activities did not differ much. Carbon sequestration parameters, soil organic carbon and basal soil respiration, increased with incorporation of organic amendments, especially in zero tillage compared to conventional and deep tillage. Root parameters such as root length, root diameter and root biomass, were not influenced by tillage types. However, ZT observations were higher than DT and CT.
Open Access
Integrated nutrient management in forage cowpea with application of liquid microbial inoculants
(Punjab Agricultural University, Ludhiana, 2019) Ramya S.; Pandove, Gulab
The present investigation was carried out to evaluate the role of liquid microbial inoculants in integrated nutrient management of forage cowpea (Vigna unguiculata). For the formulation of liquid microbial inoculants, Bacterial isolate 1 (CP-AK-SPS-4) and Bradyrhizobium sp. (NAIMCC-B-00260) were obtained from Electron Microscopy and Nanoscience Laboratory, Department of Soil Science, Punjab Agricultural University, Ludhiana and Bacterial isolate 2 (Ak-PSB-3-WHITE) was obtained from Department of Microbiology, Punjab Agricultural University, Ludhiana. Bacterial isolate 1 (CP-AK-SPS-4) and Bacterial isolate 2 (Ak-PSB-3-WHITE) were identified as Burkholderia seminalis and Burkholderia sp by sequencing of 16S rRNA gene. Further all the three cultures were evaluated for plant growth promoting features. The quantity of IAA, phosphate solubilization and gibberellic acid produced by Burkholderia seminalis was 7.05 ± 0.89 μg/ml, 137.57 ± 4.66 μg/ml and 160.73 ± 9.49 μg/ml respectively, by Burkholderia sp. was 5.37 ± 0.54 μg/ml, 130.28 ± 6.63 μg/ml and 129.94 ± 6.49 μg/ml respectively and by Bradyrhizobium sp. was 6.45 ± 0.54 μg/ml, 134.48 ± 2.78 μg/ml and 118.96 ± 3.51 μg/ml respectively. Siderophore production was exhibited only by Burkholderia seminalis (134.8 ± 4.80 μg/ml catechol type and 145.4 ± 5.09 μg/ml hydroxamate type). Liquid microbial inoculants of Burkholderia seminalis, Burkholderia sp. and Bradyrhizobium sp. were prepared using trehalose (5mM) in basal medium and showed maximum viable count upto 180 days of storage as compared to charcoal carrier based formulations at room and refrigerated temperature. The field experiment was conducted at Punjab Agricultural University, Regional Research Station, Bathinda and Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana during Kharif 2018 respectively. A total of eleven treatment combinations of liquid microbial inoculant (Burkholderia seminalis, Burkholderia sp. and Bradyrhizobium sp.) with 75% and 100% RDF were framed in randomized block design and replicated thrice. It was observed that treatment T10 (75%RDF + Burkholderia sp. + Burkholderia seminalis) significantly (p<0.05) improved vine length, stem girth, leaf length, leaf breadth, number of branches per plant, number of leaves per plant, leaf: stem ratio, green fodder yield, dry matter yield, acid detergent fiber (ADF), neutral detergent fiber (NDF), crude protein content, in vitro dry matter digestability (IVDMD), total ash content, total chlorophyll content, total sugars content and total phenolic content. Moreover, percentage increase in green fodder yield (q/ha) with T10 treatment over T1 (control) was 12.86%. Scanning electron microscopy provided insight of root colonization of forage cowpea by liquid microbial inoculants under gnotobiotic conditions, maximum root surface colonization was observed by liquid microbial inoculants of Burkholderia seminalis followed by Burkholderia sp. and Bradyrhizobium sp. which further strengthened the results of field experiments. Therefore, it can be concluded that liquid microbial inoculants could play a predominant role in integrated nutrient management of forage cowpea for enhanced productivity and quality.

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