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Open Access
Comparative study on phosphate solubilization potential of bacteria in wheat under agroforestry vs open cultivation systems
(G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-10) Hedi, Anjali; Sahgal, Manvika
To fulfill the rising need for food, chemical fertilizers and pesticides have been used excessively, which has surely resulted in pollution and significant harm to soil fertility. The use of bio fertilizers, an environmentally benign agricultural input that is also more affordable than chemical fertilizers, significantly increases soil fertility over time. Phosphate solubilizing bacteria associated with plant increase the plant available phosphorus in soil thereby promoting the plant growth as well as yield. The current study examined the plant growth promoting potential of two PSB Pseudomonas strains on wheat through a field trial in open cultivation system and agroforestry system. Both the strains were previously isolated from the Dalbergia sissoo forest in the North-western Himalayan region of India and tested for their ability to serve as bio fertilizer using an integrative approach that included field trial with recommended dose of NPK. They were identified as Pseudomonas paralactis L3 (NCBI Gen Bank Accession number MG966341), Culture Accession number NAIMCC-B-03170 and Pseudomonas aeroginosa P2 (NCBI Gen Bank Accession number MG966347), Culture Accession number NAIMCC-B-03171). The field trial was conducted in two experimental setup i.e. open cultivar with Randomized Block Design (RBD) having three replicates per treatment and in all three treatments,P2, L3 individual and in consortium, CNS (L3+P2). At 60DPI and 120DPI in open field cultivation, CNS showed the best result in all agronomic parameters, GY, BY, HI and 1000 grain weight, photosynthetic pigments, microbial count and soil enzymatic activities and were significantly higher than untreated control at p<0.05. Whereas in agroforestry system P2 showed the best result. At 120DPI the highest GY (14.37qha-1), BY(16.27qha-1), HI (88.29) and1000grain yield (36g) was observed in P2 strain in agroforestry system. Whereas in open cultivation system GY (13.27qha-1), BY(15.06qha-1), HI (88.1) and 1000grain yield (33.33g) was shown by CNS. The highest NPKcontent in soil during open field cultivation was observed in CNS (37.30%,0.44% and 1.63%) and in agroforestry system the maximum response was observed in P2 (38.09%, 0.482% and 1.69%) as compared to uninoculated control. In open system the highest protein and carbohydrates content were found in CNS10.82% and 2.603% whereas in agroforestry system P2 showed the highest response 12.52% and 2.93%. To assess the impact of PSB on wheat seed germination the in vivo seed germination assay was performed in which P2 treated seeds showed significantly higher seed shoot length (SL) crawling as compared to untreated wheat seedling. Overall while comparing both the systems the best results were obtained in agroforestry system as compared to open field cultivation.
Open Access
Biodegradation of cypermethrin and profenofos using indigenous bacteria and their subsequent influence on microbial dynamics of rhizospheric soil of wheat and maize
(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-07) Singh, Harshita; Sharma, Anita
Cypermethrin and Profenofos degrading bacterial isolates were recovered from the pesticide contaminated site of agricultural fields of U.S. Nagar, Uttarakhand. Recovered bacterial isolates were enriched to grow in Minimal Salt Medium in the presence of Cypermethrin and profenofos (20 to 100 ppm). Two bacterial isolates (HS2A and HS2B) were selected for the study on the basis of their ability to degrade and utilize Cypermethrin and Profenofos as a source of Carbon and phosphorus. Both the isolates tolerated 100ppm of Cypermethrin and Profenofos. Bacterial isolates were characterized on the basis of morphological, biochemical and molecular tests. HS2A was characterized as Bacillus amyloliquifaciens and HS2B with Staphylococcus gallinarum showed more than 95% s e q u e n c e homology. HS2A and HS2B biodegraded Cypermethrin and Profenofos (100ppm) in Minimal Salt Medium. After 36 h, the pattern of Cypermethrin and Profenofos degradation was 88.9% > 76.31% and 84.62% > 79.03% by HS2A and HS2B respectively. Degradation of both the pesticides in soil slurry was 61.54% > 61.21% and 54.38%, 67.01% > 66.80% and 63.86% by mixture of HS2A+HS2B and HS2A, HS2B respectively after 15 days. HS2A also decolorized toluidine blue in MSM. Pot experiment conducted on maize and wheat using unsterilized soil spiked with Cypermethrin (0.806 ml/100ml water) and Profenofos (0.714 ml/100ml water) showed best performance in consortium treatment. GC-MS analysis of intermediates of biodegraded Cypermethrin and Profenofos with bacterial isolates did not show the presence of toxic metabolites after 36 hours in Minimal Salt Medium. Bacterial isolates degraded Cypermethrin and Profenofos into Benzene, 1,3-bis(1,1-dimethylethyl)-, Dodecane, 4,6-dimethyl-, Dodecane, n-Hexadecanoic acid, Eicosane, 2,4-dimethyl- and 13-Hexyloxacyclotridec-10-en-2-one Hexane, 3,3,4- trimethyl. Soil health and microbial diversity were better in treated soil than control.
Open Access
Exploration of zinc solubilizing bacteria and their prolific contribution in zinc biofortification of rice grains
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-12) Upadhayay, Viabhav Kumar; Singh, Ajay Veer
The current study was an endeavor to characterize zinc-solubilizing bacteria, bearing massive plant growth-promoting traits, from the rhizosphere of millets (barnyard millet and finger millet). In addition, the study was conducted to evaluate the effect of potential bacterial strains and their consortia on growth and the biofortification benefits by assessing improved level of zinc assimilation in rice under pot and field conditions. A total of 130 bacteria were isolated and tested for zinc solubility characteristics, of which only 10 bacterial isolates were selected based on their zinc solubilization potential. Finally, 3 bacterial isolates (BMAR64, FMAR105 and BMRR126) were selected based on the quantitative pattern of zinc solubility potential and their compatibility with one another, and were used for the development of bacterial consortia. These bacteria solubilized Zn profoundly determined quantitatively in liquid broth by AAS and SEM-EDX. These zinc-solubilizing bacterial strains were characterized on the basis of morphological, biochemical tests and 16S rDNA sequencing. These isolates BMAR64, FMAR105 and BMRR126 were recognized as Pantoea rodasii, Serratia marcescens and Burkholderia cepacia, respectively, and they were also found to have genes for organic acid production (gcd and gdh). All three bacterial isolates and their consortia were evaluated for their effect on rice plants (variety- Pusa Basmati -1) in a pot trial in which the treatment with bacteria and their consortia with ZnO supplementation established narrative of Zn-biofortification and improvement in the growth of rice plant. The zinc solubilizing bacterial consortia (Consortium 1 and Consortium 2) with ZnO supplementation exhibited the maximum Zn content in grains of rice (variety- Pusa Basmati-1) in both the Terai and Katchar region of Northern part of India. The treatments containing single bacterial inoculants (BMRR126) with 60% RDF and Consortium (BMAR64+FMAR105+BMRR126) with 100% RDF with ZnO amendment exhibited promising level of Zn content in grain of rice plant (variety- Pant Dhan 18) grown in Terai region. All treatments containing zinc-solubilizing bacterial strains increased rice crop productivity in terms of several aspects such as plant height, dry matter accumulation, number of tillers and effective tiller, grain yield, straw yield, biological yield, panicle length, number of grains per ear, and thousand grain weight. The soil parameters (pH, EC, organic carbon, NPK, available Zn, and soil dehydrogenase activity) also positively influenced under all bacterial treatments compared to uninoculated control. The present study gives an impression of the use of zinc solubilizing bacteria and their consortia as effective bioinoculants (supplementation with economic feasible source of zinc, i.e. ZnO) to harness zinc biofortification of rice crops in more ecofriendly way for sustainable agriculture. Our study clearly highlights the need for zinc-solubilizing bacteria to achieve the benefits of zinc biofortification in different regions.
Open Access
Evaluation of salt tolerant bacterial consortia on growth of green gram (Vigna radiata) under saline and non-saline conditions
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-12) Negi, Riya; Singh, Ajay Veer
Salinity is one of the most crucial problems for sustainable agriculture which is severely affecting crop growth and decreasing the food production. On the other side, the world's growing population necessitates increased food production. So, there is a need of hours to increase agricultural production from salt affected soils by adopting cost-effective and environment friendly approaches. Use of salt tolerant plant growth promoting rhizobacteria (PGPR) could be a promising option to enhance the production of crops in salt affected soils and reduce the negative impacts of salt stress in soil. In this regard, the present study was conducted with 7 Plant growth promoting bacteria, which were further rescreened for salt tolerance capacity at different NaCl concentrations. These Plant growth promoting bacteria were capable to tolerate 15-20% of NaCl concentrations. Further, On the basis of biocompatibility assay, 3 Consortia i.e. Consortium1 (SRK14 and MRCII6), Consortium C2 (RRCI5 and SRP9II1) and Consortium C3 (CP17 and CP20) were prepared. All the bacterial isolates and consortia were screened for different plant growth promoting traits such as indole-3- acetic acid (IAA), Siderophore production, K solubilization, P solubilization, Zinc solubilization, Ammonia production, N2 fixation and exopolysaccharide production. Based on these results, bacterial consortia showed better PGPs traits as compare to single bacterial isolates. Therefore further selected for towel paper assay and pot experiment on green gram (Vigna radiata var. PM9) under saline and non-saline conditions. In paper towel assay, a significant enhancement of root length, shoot length, standard germination, first count, plant fresh weight, dry weight, seedling vigour index I, seedling vigour index II, relative growth index, germination value, mean daily germination and peak value were recorded in plants inoculated with bacterial consortia under both saline and non saline conditions over uninoculated plants. Similarly, in pot trial assay, at elevated concentrations of NaCl, the adverse effects on plant height, no. of nodules, chlorophyll content, carotenoid content, no. of pods per plant, no. of seeds per pod, 100 grains weight along with yield attributing characteristics were recorded in uninoculated (only NaCl) plants. However, in similar conditions inoculation of bacterial consortia showed positive adaption and improve growth performance of green gram compared to uninoculated. Hence, the results suggested that plant inoculated with Consortium 1(SRK14 and MRCII6) under non saline conditions and under saline conditions Consortium C3 (CP17 and CP20) showed better results over uninoculated control. Therefore, Consortium C1 (SRK14 and MRCII6) under non saline conditions is a potential consortium for promoting plant growth attributes in green gram and Consortium C3 (CP17 and CP20) under saline conditions is a potential salt tolerant consortium to mitigate the deleterious effects of salinity in green gram.
Open Access
Isolation of cellulose degrading microorganisms from biogas slurry and their application in rice straw degradation
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-10) Deorari, Pinki; Sahgal, Manvika
Rice is one of the major food grain crops in the world. Besides grain, it also produces large amount of waste in the form of rice straw. Present study explores the biogas slurry microorganisms in the biodegradation of rice straw. Efforts were also made to identify potential cellulase producing microbial cultures from biogas slurry that play an important role in the hydrolysis of lignocellulosic biomass. 45 bacterial and fungal cultures were isolated from biogas slurry. These isolates were screened for cellulase and xylanase enzyme activities both qualitatively and quantitatively. The potential fungal isolate SSF4 exhibiting maximum FPase (0.315 U/mL) activity on CMC minimal salt medium, fungal isolate SSF1 showing highest β-glucosidase (0.552 U/mL) activity on CMC minimal salt medium and SSF1 showed maximum xylanase (0.510 U/mL) activity on wheat bran minimal salt medium were selected for biodegradation of rice straw. Based on the morphological characterization and microscopic observation of selected strain revealed that SSF1 culture was identified as Aspergillus genus and SSF4 culture as Trichderma genus. The consortium of these two strains with biogas slurry was effective in biodegradation of rice straw. The rice straw was first subjected to alkaline pretreatment for removal of lignin content from rice straw. For biological treatment, the alkaline pretreated rice straw was treated with selected fungal isolates and biogas slurry. The pretreated and biologically treated rice straw was monitored through FTIR analysis to reveal the chemical changes in the lignocellulosic biomass of rice straw. The future research should be focus on the study this consortium at the biochemical level and optimization would be needed for higher production of enzyme to improve biodegradation of rice straw.
Open Access
Bioremediation of polluted Kalyani river water using immobilized microbial consortia
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-10) Rawal, Himanshu; Tewari, Lakshmi
The pollution of rivers and other water bodies has become one of major concern for the entire world especially for the developing nations like India. The effluents of various industries, loaded with synthetic dyes in large amount, are great threat for the ecosystem and their native flora and fauna. Therefore, the present investigation was carried out for the bioremediation of several toxic industrial azo dyes (Congo red, Direct brown and Diazo black) and wastewater from Kalyani river, Rudrapur, Uttarakhand, using eighteen microbial cultures (fungi, microalgae and cyanobacteria). All the microbial cultures were initially screened for their in-vitro dye decolorization efficiencies. Then, the selected isolates were further evaluated spectrophotometrically for the quantification of dye decolorization for all three azo dyes. Five fungal cultures, (WRTP2, WRTP3, WRTP4, BS1 and WRFP1) were able to decolorize all the three azo dyes in 1-4d. The microbial consortia were developed using the potential microbial monocultures (fungi, microalgae and cyanobacteria) to increase the extent of dye removal for the three azo dyes (Congo red, Direct brown and Diazo black). The consortia were more efficient and showed complete decolorization of the liquid cultures in lesser time (within 48h). The effect of dye concentration and pH of the medium were also found to have significant impact and the developed microbial consortia (WRTP3+Syn) and (BS1+ Syn) were able to grow and decolorize the azo dyes (Congo red and Direct brown) upto a conc. of 1200 mgL-1. The immobilized co-cultures (on stainless steel sponge and cotton plugs) showed rapid decolorization of dye direct brown and were able to decolorize 100 mgL-1 and 200 mgL-1 of dye in 14h and 20h respectively. The extent of dye decolorization during microbial decolorization process was monitored by UV-visible spectroscopy, FT-IR and SEM analysis. The Kalyani river water was analysed for its physio-chemical parameters. Reduction in the values of BOD, COD, electrical conductivity, pH, colour and turbidity in the microbially treated wastewater sample was also recorded.
Open Access
Bioconversion of pretreated paddy biomass into sugar using combined chemical and microbial approach
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-10) Solanki, Anjali; Tewari, Lakshmi
Rice is the major staple food of our county and most part of the world.. Rice waste generation and their open field burning is major problem, resulting in nutrient losses as well as major air quality issues that threaten human health and safety. Therefore the present investigation was carried for the conversion of these paddy waste into valuable fermentable sugar. For this study 17 fungal and 5 bacterial microbial cultures were isolated from Crop research centre, Pantnagar, Uttarakhand and some cultures were collected from Department of Microbiology. Then, the selected isolates were further screened qualitatively and quantitavely for their extracellular enzyme activities i.e. cellulase, laccase, silicase with the aim to select cultures of best enzyme activities for removal of lignin and silica content from rice straw biomass and their conversion into fermentable sugar. In this study, various pretreatments such as chemical and biological were given to remove silica and lignin content from rice straw. Microbial culture RSFP3 was found to give maximum FPase enzymatic activity (0.108U/ml) after 7th day of incubation period which was isolated from rice stem. Fungal culture RSFP1 give maximum silicase enzyme activity (276.62mg/L) after 10th day of incubation period. Fungal cultures BS6 reported to give maximum laccase enzyme activity (115.40U/ml) after 12th day of incubation period. On the basis of given results microbial cultures RSFP1 and BS6 were selected for further study to remove silica and lignin content from rice straw. Consortium of fungal isolate (RSFP1+BS6) remove the lignin content from 14.65% to 9.3% and % silicon solubilisation by using these consortium is 92.79mg/L. After the pretreatments treated biomass was further used for conversion into fermentable sugar by using fungal culture PPCF.
Open Access
Functional and molecular characterization of rhizobacterial antagonists against foliar blight pathogens of wheat, Bipolaris sorokiniana and Alternaria triticina
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-02) Jaggi, Vandana; Sahgal, Manvika
Foliar blight pathogens affect the wheat production and growth worldwide. The application of chemical fertilizers to control the disease has harmful effects to human health and environment. Fortunately, rhizobacteria can be used as an alternative strategy for the sustainable disease management. In present study, a total of forty five rhizobacteria isolated from rhizosphere soil of wheat cultivated in five agroclimatic regions across four India states were tested for in vitro antagonistic activity against two foliar blight pathogens; Bipolaris sorokiniana and Alternaria triticina causing wheat spot blotch and leaf blight respectively. In addition, all 45 rhizobacteria were tested for production of hydrolytic enzymes (chitinase, protease, cellulase, amylase, and lipase) and PGP traits (solubilization of phosphate & zinc, production of ammonia and siderophore). Six isolates scoring (Ʃ=>12) on bonitur scale were selected for further studies. The isolates P10, UP11, MH13, MP17, MH12, and MP14 were subsequently identified as Bacillus methylotrophicus (MN099430.1), Bacillus subtilis (MN099431.1), Bacillus sp. (MN099432.1), Streptomyces sp. (MN099435.1), Lysinibacillus sp. (MN099433.1), and Staphylococcus epidermidis (MN099434.1) respectively. In vivo pot trial revealed that seed bacterization plus booster dose of bacterial treatments resulted in lowest disease incidence (DI %) than foliar spray 24 hrs after pathogen infection against both the foliar blight pathogens. Reduction in spot blotch incidence in wheat plants treated with the isolates P10 (T2) UP11 (T3) was significant with48.66% and 50.33 % DI respectively. Likewise, percrent reduction in the incidence of leaf blight by P10 and UP11 was 49.26% and 50.33% respectively. The pot experiments also revealed that P10 and UP11 considerably increased plant biometric parameters. Therefore, B. methylotrophicus strain P10 and B. subtilis strain UP11 were equally effective to control both the foliar blight pathogens individually as well as in a complex. Antifungal metabolites produced by P10 and UP11 were analyzed in ethyl acetate extract by GC-MS and 29 and 30 compounds were identified respectively. A compound, isobutylhexahydropyrrolo [1, 2-a] pyrazine-1, 4-dione was dominant and abundant in both the strains. In silico studies revealed a strong interaction (ΔG-6.76) between the dominant compound and, ceratoplatinin, a phytotoxic protein of fungus B. sorokiniana. Hence, we suggest that strain P10 and UP11 has the potential to be used in sustainable agriculture.
Open Access
Bioconversion of photooxidatively delignified woody biomass of Salix alba into bioethanol using C6/C5 co- sugar fermenting yeast Pichia kudriavzevii BGY1
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-03) Bisht, Neha; Tewari, Lakshmi
Presently bioethanol is considered as the most sustainable and eco- friendly fuel in contrast to non- renewable fuels. The present study was carried out for identification and characterization of potential ethanogenic yeast. Native yeast cultures were isolated from diverse fruit sources. Based on qualitative and quantitative screening for ethanol production two ethanogenic yeast were selected and subjected to UV and gamma mutagenesis with aim to enhance stress tolerance limit and sugar fermenting ability. Based on phenotypic and 5.8 S ITS rRNA gene sequencing, the isolate BGY1 and its mutant BGY1- γ m were identified as Pichia kudriavzevii BGY1 and Pichia kudriavzevii BGY1- γ m. The woody biomass of Salix alba was first subjected to chemical, biological, chemi- biological and photonanocatalyst mediated pretreatments for removing lignin and improving saccharification efficiency. Combined photonanocatalyst induced and cellulase enzyme aided alkaline oxidative pretreatment decreased the lignin content from 18.7 to 4.0 %. The pretreated substrate was then saccharified into fermentable sugars using cellulase enzyme from the fungal isolate (Aspergillus terrus PPCF). Optimization of saccharification resulted in 57.2 % of saccharification efficiency in 24 h. Then, the potential microbial strains in mono/co-cultures were screened for their alcohol producing efficacies using saccharified Salix alba wood biomass. Bio-conversion of saccharified wood employing potential strains into bio-ethanol was carried out in batch cultures through SHF and SSF process. Finally the co- culture of P. Kudriavzevii BGY1 and P. Kudriavzevii BGY1-γ m were selected for alcoholic fermentation. Alcoholic fermentation of saccharified Salix alba wood powder using co- culture of yeast P. kudriavzevii BGY1 and P. kudriavzevii BGY1- γ m at 32º C under SSF batch fermentation process was found superior over batch fermentation under SHF process. Under optimized conditions maximum ethanol yield 13.0 % was achieved with saccharified Salix alba wood.

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