Metabolite profiling of plant growth promoting cyanobacteria-Anabaena laxa and Calothrix elenkinii, using untargeted metabolomics.

The metabolite profiles of two plant growth promoting cyanobacteria-Anabaena laxa and Calothrix elenkinii, which serve as promising biofertilizers, and biocontrol agents were generated to investigate their agriculturally beneficial activities. Preliminary biochemical analyses, in terms of total chlorophyll, total proteins, and IAA were highest at 14 days after inoculation (DAI). In A. laxa 20-25% higher values of reducing sugars, than C. elenkinii at both 14 and 21 DAI were recorded. Carbon and nitrogen assimilating enzyme activities-phosphoenol pyruvate carboxylase (PEPC), carbonic anhydrase (CA), and glutamine synthetase (GS) were highest at 14 DAI, albeit, nitrate reductase (NR) activity was higher by 0.73-0.84-fold at 21 DAI. Untargeted GC-MS (Gas chromatography-Mass spectrometric) analysis of metabolite profiles of 21d-old cyanobacterial cultures and characterization using NIST mass spectral library illustrated that A. laxa recorded highest number of metabolite hits in three chemical classes namely amino acid and peptides, nucleotides, nucleosides and analogues, besides other organic compounds. Based on the pathway analysis of identified metabolites, both A. laxa, and C. elenkinii were enriched in metabolites involved in aminoacyl-tRNA biosynthesis, and amino acid metabolism pathways, particularly lactose and glutamic acid, which are important players in plant-microbe interactions. Correlation-based metabolite network illustrated distinct and significant differences in the metabolic machinery of A. laxa and C. elenkinii, highlighting their novel identity and enrichment in C-N rich metabolites, as factors underlying their plant growth and soil fertility enhancing attributes, which make them valuable as inoculants. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03902-7.

Enhancing physiological metrics, yield, zinc bioavailability, and economic viability of Basmati rice through nano zinc fertilization and summer green manuring in semi-arid South Asian ecosystem.

During the summer and rainy seasons (April-October) of 2020 and 2021, two consecutive field experiments were conducted at the research farm of the ICAR-Indian Agricultural Research Institute, New Delhi, India. In this study, we examined the effects of summer green manuring crops (GM) and a variety of zinc fertilizers (ZnF) on Basmati rice (Oryza sativa L.) growth, physiological development, yield response, zinc nutrition and economic returns. A combination of GM residues and nano zinc fertilization helped significantly enhancing Basmati rice's growth and its physiological development. Following the incorporation of Sesbania aculeata (Sesbania), successive Basmati rice physiological parameters were significantly improved, as well as grain, straw, biological yields, harvest index and economic returns. The highest Zn content of 15.1 mg kg -1 and the lowest of 11.8 mg kg -1 in milled rice grain were recorded in Sesbania green manuring (G2) and control i.e., in the fallow (G1), respectively. Coating onto urea with 0.2% nano zinc oxide (NZnCU) was observed to be more effective than other zinc sources in terms of growth parameters, yield attributes, zinc nutrition, grain and straw yields for succeeding Basmati rice crop; however, the effects were comparable to those of bulk zinc oxide-coated urea (BZnCU) of 1%. The highest Zn content of 15.1 mg kg -1 was recorded with the application of 1% BZnCU and the lowest of 11.96 mg kg -1 with the soil application of 5 kg Zn ha -1 through bulk ZnO in the milled rice grain. Application of 1% BZnCU led to a 26.25% increase in Zn content of milled rice grain compared to soil application of 5 kg Zn ha -1 through bulk ZnO. As a result, the combination of inclusion of Sesbania aculeata (Sesbania) residue and 0.2% NZnCU was identified as the most effective treatment, for Basmati rice growth and physiological development. A combination of nano Zn fertilization in conjunction with the incorporation of green manure can be advocated for better growth, physiological performance, zinc dense grains, and higher profitability of Basmati rice for farmers and consumers.

Elevated CO2 along with inoculation of cyanobacterial biofilm or its partners differentially modulates C-N metabolism and quality of tomato beneficially.

Diazotrophic cyanobacteria are known to influence nutrient availability in soil, however, their benefits under elevated CO2 environment, particularly on fruit quality attributes, is a less investigated aspect. Laboratory developed cyanobacterium-fungal biofilm (An-Tr), composed of Anabaena torulosa (An) as the matrix with the partner as Trichoderma viride (Tr), along with the individual partners were evaluated under ambient (aCO2-400 ± 50 ppm) and elevated (eCO2-700 ± 50 ppm) conditions, with and without tomato plants. An-Tr inoculation exhibited distinct and significantly higher values for most of the soil microbiological parameters, plant growth attributes and antioxidant/defense enzyme activities measured at 30 and 60 DAI (days after inoculation). Significant enhancement in soil nutrient availability, leaf chlorophyll, with 45-50% increase in the enzyme activities related to carbon and nitrogen assimilation, higher yields and better-quality parameters of tomato, with An-Tr biofilm or An inoculation, were recorded, particularly under eCO2 conditions. The fruits from An-Tr treatments under eCO2 exhibited a higher titrable acidity, along with more ascorbic acid, carotenoids and lycopene content, highlighting the superiority of this inoculant. Multivariate analyses revealed significant (p ≤ 0.05) interactions among cultures, DAI, and CO2 levels, illustrating that cyanobacterial inoculation can be advocated as a strategy to gainfully sequester eCO2. Significant improvement in yield and fruit quality along with 50% N savings, further attest to the promise of cyanobacterial inoculants for tomato crop in the climate change scenario.

Azospirillum-Bacillus associations: synergistic effects on in vitro PGP traits and growth of pearl millet at early seedling stage under limited moisture conditions.

The association of plant beneficial Azospirillum and Bacillus spp. strains expressing different sets of PGP traits may have complementary or supplementary effects on host plants. In the present investigation, A. formosense and Bacillus spp. strains showing diverse PGP traits (IAA production, nitrogenase activity, phosphate, zinc and potassium solubilization, siderophores, antagonism against phytopathogens, osmotic stress tolerance, etc.) were assessed for compatibility by cross-streaking and co-culturing. Under co-culture (Azospirillum + Bacillus), a significant increase in the expression of PGP traits, nitrogenase activity (up to 89%), phosphate solubilization (upto 236%), siderophore production (upto 20%) was observed as compared to individual Azospirillum culture, indicating synergistic effect of co-culture. IAA production was higher in Azospirillum sp. strains as compared to Bacillus spp. strains, when cultured individually; however, when co-cultured, the IAA levels were in the mid-range indicating the contributory effects of compatible strains. The effect of individual Azospirillum and Bacillus strains and their co-inoculation was also assessed on the growth of pearl millet at early stages under moisture-deficit stress imposed using PEG6000 (0, 10, and 20%). Co-inoculation enhanced seed germination (up to 10, 3, and 6% increase under 0, 10, and 20% PEG, respectively, over individual Azospirillum treatment), root traits (increased root hair density and lateral branches), and seedling vigor indices (up to 22, 32, 43% increase in seed vigor index I and 8, 14, and 10% increase in seed vigor index II under 0, 10, 20% PEG, respectively, over individual Azospirillum treatment) under normal as well as moisture-deficit conditions suggesting the role of Bacillus spp. strains in better adaptation of the plants to stress and higher yield potential. The synergistic effect of co-cultured Azospirillum and Bacillus strains on PGP traits indicated metabolic interplay between the two strains which needs to be further understood. The positive effect of co-inoculation on plant growth under moisture-deficit stress indicated the promise of Azospirillum and Bacillus as a synergistic bioformulation for combating nutrient and drought stress in pearl millet, particularly in nutrient-poor dryland agricultural systems. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03503-4.

Differential effects of rhizobacteria from uninfected and infected tomato on Meloidogyne incognita under protected cultivation.

Intermingled uninfected and root-knot nematode-infected tomato plants are commonly observed under protected cultivation. To understand the role of rhizobacteria underlying the susceptibility to nematode infectivity in these tomato plants, 36 rhizobacteria (18 from each type) with morphologically distinct colony characteristics were isolated from the rhizosphere of uninfected and root-knot nematode-infected tomato plants. The in vitro nematicidal potential of rhizobacteria from the uninfected rhizosphere was significantly higher than that from the infested rhizosphere. The three most effective antagonists were identified as Microbacterium laevaniformans, Staphylococcus kloosii, Priestia aryabhattai from root-knot-nematode-infected tomato rhizosphere and Staphylococcus sciuri, Bacillus pumilus, and Priestia megaterium from the rhizosphere of uninfected tomato. Volatile organic compounds from these rhizobacteria were characterized. Except for S. kloosi, the soil drenching with other rhizobacteria significantly reduced juvenile penetration (>60%) in tomato roots. Furthermore, the application of a single or consortium of these rhizobacteria affected nematode reproduction in tomato. Four consortia of rhizobacteria (S. sciuri + B. pumilus + P. megaterium), (B. pumilus + P. megaterium), (S. sciuri + B. pumilus), and (S. sciuri + P. megaterium) from uninfested rhizosphere and two consortia (M. laevaniformans + P. aryabhattai), (M. laevaniformans + S. kloosii + P. aryabhattai) from infested rhizosphere (IRh) effectively reduced M. incognita reproduction and considerably enhanced plant growth and yield in tomato. The nematicidal efficacy, however, decreased when S. kloosii was applied in the consortium. These distinctive effects illustrate how the plant susceptibility to nematode infectivity is modulated under natural conditions.

Untargeted GC-MS reveals differential regulation of metabolic pathways in cyanobacterium Anabaena and its biofilms with Trichoderma viride and Providencia sp.

Cyanobacteria and their biofilms are used as biofertilizing options to improve plant growth, soil fertility, and grain quality in various crops, however, the nature of metabolites involved in such interactions is less explored. The present investigation compared the metabolite profiles of cyanobacterial biofilms: Anabaena torulosa- Trichoderma viride (An-Tr) and A. torulosa- Providencia sp. (An-PW5) against the individual culture of A. torulosa (An) using untargeted gas chromatography-mass spectroscopy. Metabolites were identified using the NIST mass spectral library and the relative peak area of cultures analysed, after normalization with an internal standard, ribitol. An-Tr biofilm recorded approximately 66.85% sugars, with increased quantity and numbers of sugars and their conjugates, which included maltose, lactose, and d-mannitol, but decreased amino acids concentrations, attributable to the effect of Tr as partner. Heat map and cluster analysis illustrated that An-Tr biofilm possessed a unique cluster of metabolites. Partial least square-discriminate analysis (PLS-DA) and pathway analyses showed distinct modulation in terms of metabolites and underlying biochemical routes in the biofilms, with both the partners- PW5 and Tr eliciting a marked influence on the metabolite profiles of An, leading to novel cyanobacterial biofilms. In the An-PW5 biofilm, the ratios of sugars, lactose, mannitol, maltose, mannose, and amino acids serine, ornithine, leucine and 5­hydroxy indole acetic acid were significantly higher than An culture. Such metabolites are known to play an important role as chemoattractants, facilitating robust plant -microbe interactions. This represents a first-time study on the metabolite profiles of cyanobacterial biofilms, which provides valuable information related to their significance as inoculants in agriculture.

Palladium encapsulated nanofibres for scavenging ethylene from sapota fruits.

Scavenging ethylene is a useful intervention during the transportation and storage of tropical climacteric fruits like sapota. Sapota (Manilkara achras Mill.) is a delicious tropical fruit with a very high respiration rate and poor shelf life. To prolong its post-harvest shelf life, the use of palladium chloride in electrospun nanomats was evaluated at a concentration varying from 1 to 4% levels. Encapsulation of 1-2% PdCl2 in nanomats increased the ethylene scavenging capacity (ESC) by 47-68%. Although, upon encapsulation, both PdCl2 and potassium permanganate showed significantly the same ethylene scavenging activity, the efficacy of PdCl2 was found better in presence of sapota fruits. The PdCl2 nanomats were brighter (L* > 73) in colour compared to the potassium permanganate mat. The placement of nanomats (2 cm2 × 9 cm2) in corrugated fibre board boxes in which the sapota was packed showed higher quality indices (firmness, TSS, ascorbic acid, and phenolics) along with lower PLW and respiration rate during the 8 days of storage period. Compared to control (8.35%), physiological loss in weight of 4.47% was recorded in fruits stored with ethylene scavenging nanomats. PdCl2 encapsulated PVA nanomats can emerge as a promising option for the retention of quality in fruits during storage and transit.

Prospecting the significance of methane-utilizing bacteria in agriculture.

Microorganisms act as both the source and sink of methane, a potent greenhouse gas, thus making a significant contribution to the environment as an important driver of climate change. The rhizosphere and phyllosphere of plants growing in natural (mangroves) and artificial wetlands (flooded agricultural ecosystems) harbor methane-utilizing bacteria that oxidize methane at the source and reduce its net flux. For several decades, microorganisms have been used as biofertilizers to promote plant growth. However, now their role in reducing net methane flux, especially from flooded agricultural ecosystems is gaining momentum globally. Research in this context has mainly focused on taxonomic aspects related to methanotrophy among diverse bacterial genera, and environmental factors that govern methane utilization in natural and artificial wetland ecosystems. In the last few decades, concerted efforts have been made to develop multifunctional microbial inoculants that can oxidize methane and alleviate greenhouse gas emissions, as well as promote plant growth. In this context, combinations of taxonomic groups commonly found in rice paddies and those used as biofertilizers are being explored. This review deals with methanotrophy among diverse bacterial domains, factors influencing methane-utilizing ability, and explores the potential of novel methane-utilizing microbial consortia with plant growth-promoting traits in flooded ecosystems.

Influence of cyanobacterial inoculants, elevated carbon dioxide, and temperature on plant and soil nitrogen in soybean.

Climate change affects nitrogen dynamics in crops and diazotrophic microorganisms with carbon dioxide (CO2 ) sequestering potential such as cyanobacteria can be promising options. The interactions of three cyanobacterial formulations (Anabaena laxa, Calothrix elenkinii and Anabaena torulosa-Bradyrhizobium japonicum biofilm) on plant and soil nitrogen in soybean, were investigated under elevated CO2 and temperature conditions. Soybean plants were grown inside Open Top Chambers under ambient and elevated (550 ± 25 ppm) CO2 concentrations and elevated temperature (+2.5-2.8°C). Interactive effect of elevated CO2 and cyanobacterial inoculation through A. laxa and Anabaena torulosa-B. japonicum biofilm led to improved growth, yield, nodulation, nitrogen fixation, and seed N in soybean crop. Nitrogenase activity in nodules increased in A. laxa and biofilm treatments, with an increase of 55% and 72%, respectively, over no cyanobacterial inoculation treatment. Although high temperature alone reduced soil microbial biomass carbon, dehydrogenase activity, and soil available N, the combined effect of CO2 and temperature were stimulatory; cyanobacterial inoculation further led to an increase under all the conditions. The highest seed N uptake (758 mg plant-1 ) was recorded with cyanobacterial biofilm inoculation under elevated CO2 with control temperature conditions. The positive interactions of elevated CO2 and cyanobacterial inoculation, particularly through A. laxa and A. torulosa-B. japonicum biofilm inoculation highlights their potential in counteracting the negative impact of changing climate along with enhancing plant and soil N in soybean.

Mycorrhizae Helper Bacteria: Unlocking Their Potential as Bioenhancers of Plant-Arbuscular Mycorrhizal Fungal Associations.

The dynamic interactions of plants and arbuscular mycorrhizal fungi (AMF) that facilitate the efficient uptake of minerals from soil and provide protection from various environmental stresses (biotic and abiotic) are now also attributed to a third component of the symbiosis. These are the less investigated mycorrhizae helper bacteria (MHB), which constitute a dense, active bacterial community, tightly associated with AMF, and involved in the development and functioning of AMF. Although AMF spores are known to host several bacteria in their spore walls and cytoplasm, their role in promoting the ecological fitness and establishment of AMF symbiosis by influencing spore germination, mycelial growth, root colonization, metabolic diversity, and biocontrol of soil borne diseases is now being deciphered. MHB also promote the functioning of arbuscular mycorrhizal symbiosis by triggering various plant growth factors, leading to better availability of nutrients in the soil and uptake by plants. In order to develop strategies to promote mycorrhization by AMF, and particularly to stimulate the ability to utilize phosphorus from the soil, there is a need to decipher crucial metabolic signalling pathways of MHB and elucidate their functional significance as mycorrhiza helper bacteria. MHB, also referred to as AMF bioenhancers, also improve agronomic efficiency and formulations using AMF along with enriched population of MHB are a promising option. This review covers the aspects related to the specificity and mechanisms of action of MHB, which positively impact the formation and functioning of AMF in mycorrhizal symbiosis, and the need to advocate MHB as AMF bioenhancers towards their inclusion in integrated nutrient management practices in sustainable agriculture.

Cyanobacterial inoculation as resource conserving options for improving the soil nutrient availability and growth of maize genotypes.

Harnessing the benefits of plant-microbe interactions towards better nutrient mobilization and plant growth is an important challenge for agriculturists globally. In our investigation, the focus was towards analyzing the soil-plant-environment interactions of cyanobacteria-based formulations (Anabaena-Nostoc consortium, BF1-4 and Anabaena-Trichoderma biofilm, An-Tr) as inoculants for ten maize genotypes (V1-V10). Field experimentation using seeds treated with the formulations illustrated a significant increase of 1.3- to 3.8-fold in C-N mobilizing enzyme activities in plants, along with more than five- to six-fold higher values of nitrogen fixation in rhizosphere soil samples. An increase of 22-30% in soil available nitrogen was also observed at flag leaf stage, and 13-16% higher values were also recorded in terms of cob yield of V6 with An-Tr biofilm inoculation. Savings of 30 kg N ha-1 season-1 was indicative of the reduced environmental pollution, due to the use of microbial options. The use of cyanobacterial formulations also enhanced the economic, environmental and energy use efficiency. This was reflected as 37-41% reduced costs lowered GHG emission by 58-68 CO2 equivalents and input energy requirement by 3651-4296 MJ, over the uninoculated control, on hectare basis. This investigation highlights the superior performance of these formulations, not only in terms of efficient C-N mobilization in maize, but also making maize cultivation a more profitable enterprise. Such interactions can be explored as resource-conserving options, for future evaluation across ecologies and locations, particularly in the global climate change scenario.

Nutrient removal by rice-wheat cropping system as influenced by crop establishment techniques and fertilization options in conjunction with microbial inoculation.

Nutrient uptake by the rice-wheat cropping system (RWCS) is an important indicator of soil fertility and plant nutrient status. The hypothesis of this investigation was that the rate and sources of nutrient application can differentially influence nutrient removal and soil nutrient status in different crop establishment techniques (CETs). Cropping system yield was on par in all the CETs evaluated, however, there were significant changes in soil nutrient availability and microbiological aspects. The system nitrogen (N), phosphorus (P), potassium (K) and zinc (Zn) uptake in aerobic rice system followed by zero tillage wheat (ARS-ZTW) was 15.7-17.6 kg ha-1, 0.7-0.9 kg ha-1, 7-9.8 kg ha-1 and 13.5-23.1 g ha-1 and higher than other CETs. The formulations of Anabaena sp. (CR1) + Providencia sp. (PR3) consortium (MC1) and Anabaena-Pseudomonas biofilm (MC2) recorded significantly higher values of soil chlorophyll and microbial biomass carbon and positively affected cropping system nutrient uptake and soil nutrient balance, illustrating the beneficial effect of microbial inoculation through increased supply of biologically fixed N and solubilised P. Zinc fertilization (5 kg Zn ha-1 through ZnSO4·7H2O as soil application) increased soil DTPA-extractable Zn by 4.025-4.836 g ha-1, with enhancement to the tune of 20-24% after two cropping cycles of RWCS. Our investigation recommends the need for change in the present CETs to ARS-ZTW, along with the use of microbial inoculation as a means of significantly enhancing cropping system nutrient uptake and soil nutrient status improvement.

Priming maize seeds with cyanobacteria enhances seed vigour and plant growth in elite maize inbreds.

Biofilm formation of a nitrogen-fixing cyanobacterium Anabaena torulosa with a beneficial fungus Trichoderma viride (An-Tr) was examined under laboratory conditions. A gradual enhancement in growth over A. torulosa alone was recorded in the biofilm, with 15-20% higher values in nitrogen fixation, IAA and exopolysaccharide production illustrating the synergism among the partners in the biofilm. To investigate the role of such biofilms in priming seed attributes, mesocosm studies using primed seeds of two maize inbred lines (V6, V7) were undertaken. Beneficial effects of biofilm (An-Tr) were recorded, as compared to uninoculated treatment and cyanobacterial consortium (Anabaena-Nostoc; BF 1-4) at both stages (7 and 21 DAS, days after sowing) with a significant increase of more than 20% in seedling attributes, along with 5-15% increment in seed enzyme activities. More than three- to fivefold higher values in nitrogen fixation and C-N mobilizing enzyme activities, and significant increases in leaf chlorophyll, proteins and PEP carboxylase activity were observed with V7-An-Tr biofilm. Cyanobacterial inoculation brought about distinct changes in the soil phospholipid fatty acid profiles (PLFA); particularly, significant changes in those representing eukaryotes and anaerobic bacteria. Principal component analyses illustrated the significant role of dehydrogenase activity and microbial biomass carbon and distinct elicited effects on soil microbial communities, as evidenced by the PLFA. This investigation highlighted the promise of cyanobacteria as valuable priming options to improve mobilization of nutrients at seed stage, modulating the abundance and activities of various soil microbial communities, thereby, enhanced plant growth and vigour of maize plants.

Synergistic effects of silver nanoparticles augmented Calothrix elenkinii for enhanced biocontrol efficacy against Alternaria blight challenged tomato plants.

The biocontrol efficacy of a cyanobacterium Calothrix elenkinii (Ce), silver nanoparticles (AgNPs) and their augmented complex (AgNPs-Ce) was evaluated. Foliar application of AgNPs-Ce reduced the disease severity by 47-58%, along with significant increases of 44-45%, 40-46% and 23-33% in leaf chlorophyll, carotenoid content, and polyphenol oxidase activity in the A. alternata infected tomato plants. A significant reduction in the pathogen load was recorded, both by plate counts and microscopic observations in the AgNPs, Ce and AgNPs-Ce treatments, while AgNPs-Ce also effectively reduced ergosterol content by 63-79%. Amplification using PCR-ITS primers revealed very faint bands or none in the AgNPs-Ce treated leaves, illustrating the inhibition of fungal growth. Significantly higher yield was recorded in the pathogen challenged plants receiving AgNPs-Ce, AgNPs, and Ce treatments. Higher expression of elicited antioxidant enzymes, along with enhanced plant growth attributes and lowered fungal load highlight the biocontrol potential of AgNPs-Ce treatment in A. alternata infected plants. This synergistic association can be explored as a promising biocontrol option against A. alternata challenged tomato plants under various agroclimatic conditions.

Statistical optimization of process parameters for improvement of phycobiliproteins (PBPs) yield using ultrasound-assisted extraction and its kinetic study.

The present study is mainly concerned for the development of an optimal ultrasound-assisted extraction (UAE) condition for phycobiliproteins (PBPs) from Oscillatoria sp. (BTA 170) using Taguchi methodology. Four process parameters viz. solid to liquid ratio, duty cycle, electrical acoustic intensity, and pH, for UAE were optimized using Taguchi methodology for enhanced PBPs extraction. The ratio of signal to noise (S/N) was used to compute the optimized condition required to attain a higher yield of PBPs, the average performance of individual parameter and corresponding interactive effects. The statistically significant parameters with their contribution were assessed using Analysis of variance (ANOVA). Results showed that duty cycle contributed the maximum influence (30.81%) on phycocyanin (PC) extraction followed by a solid liquid ratio (28.62%), pH (22.46%) and electrical acoustic intensity (18.10%). The highest contribution on the extraction of phycoerythrin (PE) was found from pH (33.16%), followed by duty cycle (31.57%), solid to liquid ratio (22.83%) and electrical acoustic intensity (12.45%). For extraction of allophycocyanin (APC), the duty cycle, solid to liquid ratio, pH and electrical acoustic intensity contributed 29.47, 29.07, 29.03, and 12.43% respectively. Results obtained from Taguchi methodology indicated that enhanced PC (94.10%), PE (95.20%) and APC (90.54%) can be achieved with solid-liquid ratio (0.2 g/ml), electrical acoustic intensity (16.99 w/cm2), duty cycle (75%), and pH 7 than the yield of PBPs obtained under unoptimized condition. In the present study, higher yield of PC (38.99%), PE (20.84%), and APC (11.93%) were attained with UAE compared to yield obtained from homogenized Oscillatoria sp. BTA 170 using 0.05 M phosphate buffer. Batch extraction data of PBPs under UAE was fitted well with the second order model. The values of second-order rate constant (k) were computed as 6.66 × 10-4, 64.09 × 10-4 and 1.49 × 10-4 L/mg/min for extraction of PC, PE and APC respectively. The PBPs exhibited significant antioxidant property and hydrogen peroxide scavenging activity, which were increased with the enhancement of PBPs concentration.

Improving Water and Nutrient Use Efficiency in Rice by Changing Crop Establishment Methods, Application of Microbial Inoculations, and Zn Fertilization.

A field experiment was conducted during the wet seasons of 2013 and 2014 to evaluate the effect of three rice establishment methods: viz. puddled transplanted rice (PTR), system of rice intensification (SRI) and aerobic rice system (ARS), two cyanobacteria based inoculants, viz. Anabaena sp (CR1) + Providencia sp (PR3) consortium and Anabaena-Pseudomonas (An-Ps) biofilm formulation, and zinc (Zn) fertilization on the rice yield, water productivity, and nutrient use efficiency. The yields of rough, brown and milled rice were highest in SRI, which was on par with PTR and both methods proved significantly superior to ARS in both years. The total water productivity of rough and brown rice in the first year was significantly higher in SRI. The SRI method saved 21.9% and 27.4% irrigation water over PTR, and savings in ARS were 37.4% and 50.8% in first and second year respectively, over PTR. The use of An-Ps biofilm formulation along with 75% RDN improved the agronomic use efficiency of both nitrogenous and phosphatic fertilizers applied. On the basis of the present study, it can be concluded that SRI improved rice yields and water productivity; while involvement of An-Ps biofilm formulation can be recommended for improved nutrient use efficiency.

Transcriptome profiling provides insights into regulatory factors involved in Trichoderma viride-Azotobacter chroococcum biofilm formation.

Azotobacter chroococcum (Az) and Trichoderma viride (Tv) represent agriculturally important and beneficial plant growth promoting options which contribute towards nutrient management and biocontrol, respectively. When Az and Tv are co-cultured, they form a biofilm, which has proved promising as an inoculant in several crops; however, the basic aspects related to regulation of biofilm formation were not investigated. Therefore, whole transcriptome sequencing (Illumina NextSeq500) and gene expression analyses were undertaken, related to biofilm formation vis a vis Tv and Az growing individually. Significant changes in the transcriptome profiles of biofilm were recorded and validated through qPCR analyses. In-depth evaluation also identified several genes (phoA, phoB, glgP, alg8, sipW, purB, pssA, fadD) specifically involved in biofilm formation in Az, Tv and Tv-Az. Genes coding for RNA-dependent RNA polymerase, ABC transporters, translation elongation factor EF-1, molecular chaperones and double homeobox 4 were either up-regulated or down-regulated during biofilm formation. To our knowledge, this is the first report on the modulation of gene expression in an agriculturally beneficial association, as a biofilm. Our results provide insights into the regulatory factors involved during biofilm formation, which can help to improve the beneficial effects and develop more effective and promising plant- microbe associations.

Trichoderma-Azotobacter biofilm inoculation improves soil nutrient availability and plant growth in wheat and cotton.

Microbial biofilms are gaining importance in agriculture, due to their multifaceted agronomic benefits and resilience to environmental fluctuations. This study focuses on comparing the influence of single inoculation-Azotobacter chroococcum (Az) or Trichoderma viride (Tv) and their biofilm (Tv-Az), on soil and plant metabolic activities in wheat and cotton grown under Phytotron conditions. Tv-Az proved superior to all the other treatments in terms of better colonisation, plant growth attributes and 10-40% enhanced availability of macronutrients and micronutrients in the soil, over control. Confocal and scanning electron microscopy showed that the cells attached to the root tips initially, followed by their proliferation along the surface of the roots. Soil polysaccharides, proteins and dehydrogenase activity showed several fold enhancement in Tv-Az biofilm inoculated samples. Time course studies revealed that the population of Az and Tv in the rhizoplane and rhizosphere was significantly higher with a 0.14-0.31 log colony-forming unit (CFU) increase in the biofilm-inoculated treatment in both crops. Enhancement in soil biological activities was facilitated by the improved colonisation of the biofilm, due to the synergistic association between Tv and Az. This demonstrates the utility of Tv-Az biofilm as a multifunctional plant growth promoting and soil fertility enhancing option in agriculture.

Interplay of phosphorus doses, cyanobacterial inoculation, and elevated carbon dioxide on yield and phosphorus dynamics in cowpea.

Phosphorus (P) demand is likely to increase especially in legumes to harness greater benefits of nitrogen fixation under elevated CO2 condition. In the following study, seed yield and seed P uptake in cowpea increased by 26.8% and 20.9%, respectively, under elevated CO2 level. With an increase in phosphorus dose up to 12 mg kg-1, seed yield enhanced from 2.6 to 5.4 g plant-1. P application and cyanobacterial inoculation increased the microbial activity of soil, leading to increased availability of P. Under elevated CO2 condition, microbial activity, measured as dehydrogenase, acid phosphatase, and alkaline phosphatase activities showed stimulation. Soil available P also increased under elevated CO2 condition and was stimulated by both P application and cyanobacterial inoculation. Higher P uptake in elevated CO2 condition led to lower values of inorganic P in soil. Stepwise regression analysis showed that aboveground P uptake, soil available P, and alkaline phosphatase activity of soil influenced the yield while available P, and organic and inorganic P influenced the aboveground P uptake of the crop. This study revealed that under elevated CO2 condition, P application and cyanobacterial inoculation facilitated P uptake and yield, mediated through enhanced availability of nutrients, in cowpea crop.

Microbial inoculants as plant growth stimulating and soil nutrient availability enhancing options for cucumber under protected cultivation.

Protected cultivation of vegetables is often hampered by declining nutrient availability in soil due to year-around farming, which in turn, leads to poor quality and yields, causing serious concern. Our study aimed towards evaluating the potential of novel biofilm formulations-Anabaena or Trichoderma as matrices with Azotobacter sp. as Anabaena-Azotobacter (An-Az) and Trichoderma-Azotobacter (Tr-Az) or together as Anabaena-Trichoderma (An-Tr), on the growth, physiological activities, yield, and changes in the profiles of soil microbial communities in two cultivars (cv. DAPC-6 and cv. Kian) of cucumber (Cucumis sativus). Photosynthetic pigments, evaluated as an index of growth showed two-threefold increase, while elicited activity of defense and antioxidant enzymes was stimulated; this facilitated significant improvement in the plants belonging to the inoculated treatments. Microbial biomass carbon and polysaccharides in soil enhanced by two-threefolds in treatments receiving microbial formulations. Available N in soil increased by 50-90% in An-Az and An-Tr biofilm inoculated treatments, while the availability of P and organic C content of soil improved by 40-60%, over control. PCR-DGGE profiles generated revealed signification modulation of cyanobacterial communities and cultivar-specific differences. Significant enhancement in leaf chlorophyll pigments, soil microbiological parameters and nutrient bio-availabilities along with positive correlation among the analysed parameters, and distinct profiles generated by PCR-DGGE analyses illustrated the promise of these novel inoculants for cucumber.