Further accelerating biologics development from DNA to IND: the journey from COVID-19 to non-COVID-19 programs.

The Coronavirus Disease (COVID-19) pandemic has spurred adoption of revolutionary initiatives by regulatory agencies and pharmaceutical industry worldwide to deliver therapeutic COVID-19 antibodies to patients at unprecedented speed. Among these, timeline of chemistry, manufacturing and control (CMC), which involves process development and manufacturing activities critical for the assurance of product quality and consistency before first-in-human clinical trials, was greatly reduced from typically 12-15 months (using clonal materials) to approximately 3 months (using non-clonal materials) in multiple cases. In this perspective, we briefly review the acceleration approaches published for therapeutic COVID-19 antibodies and subsequently discuss the applicability of these approaches to achieve investigational new drug (IND) timelines of ≤10 months in over 60 COVID-19 and non-COVID-19 programs performed at WuXi Biologics. We are of the view that, with demonstrated product quality and consistency, innovative approaches used for COVID-19 can be widely applied in all disease areas for greater speed to clinic.

Growth and protein response of rice plant with plant growth-promoting rhizobacteria inoculations under salt stress conditions.

Soil salinity has been one of the significant barriers to improving rice production and quality. According to reports, Bacillus spp. can be utilized to boost plant development in saline soil, although the molecular mechanisms behind the interaction of microbes towards salt stress are not fully known. Variations in rice plant protein expression in response to salt stress and plant growth-promoting rhizobacteria (PGPR) inoculations were investigated using a proteomic method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Findings revealed that 54 salt-responsive proteins were identified by mass spectrometry analysis (LC-MS/MS) with the Bacillus spp. interaction, and the proteins were functionally classified as gene ontology. The initial study showed that all proteins were labeled by mass spectrometry analysis (LC-MS/MS) with Bacillus spp. interaction; the proteins were functionally classified into six groups. Approximately 18 identified proteins (up-regulated, 13; down-regulated, 5) were involved in the photosynthetic process. An increase in the expression of eight up-regulated and two down-regulated proteins in protein synthesis known as chaperones, such as the 60 kDa chaperonin, the 70 kDa heat shock protein BIP, and calreticulin, was involved in rice plant stress tolerance. Several proteins involved in protein metabolism and signaling pathways also experienced significant changes in their expression. The results revealed that phytohormones regulated the manifestation of various chaperones and protein abundance and that protein synthesis played a significant role in regulating salt stress. This study also described how chaperones regulate rice salt stress, their different subcellular localizations, and the activity of chaperones.

Early anticoagulation in patients with stroke and atrial fibrillation is associated with fewer ischaemic lesions at 1 month: the ATTUNE study.

BACKGROUND: The optimal time to commence anticoagulation in patients with atrial fibrillation (AF) after ischaemic stroke or transient ischaemic attack (TIA) is unclear, with guidelines differing in recommendations. A limitation of previous studies is the focus on clinically overt stroke, rather than radiologically obvious diffusion-weighted imaging ischaemic lesions. We aimed to quantify silent ischaemic lesions and haemorrhages on MRI at 1 month in patients commenced on early (<4 days) vs late (≥4 days) anticoagulation. We hypothesised that there would be fewer ischaemic lesions and more haemorrhages in the early anticoagulant group at 1-month MRI. METHODS: A prospective multicentre, observational cohort study was performed at 11 Australian stroke centres. Clinical and MRI data were collected at baseline and follow-up, with blinded imaging assessment performed by two authors. Timing of commencement of anticoagulation was at the discretion of the treating stroke physician. RESULTS: We recruited 276 patients of whom 208 met the eligibility criteria. The average age was 74.2 years (SD±10.63), and 79 (38%) patients were female. Median National Institute of Health Stroke Scale score was 5 (IQR 1-12). Median baseline ischaemic lesion volume was 5 mL (IQR 2-17). There were a greater number of new ischaemic lesions on follow-up MRI in patients commenced on anticoagulation ≥4 days after index event (17% vs 8%, p=0.04), but no difference in haemorrhage rates (22% vs 32%, p=0.10). Baseline ischaemic lesion volume of ≤5 mL was less likely to have a new haemorrhage at 1 month (p=0.02). There was no difference in haemorrhage rates in patients with an initial ischaemic lesion volume of >5 mL, regardless of anticoagulation timing. CONCLUSION: Commencing anticoagulation <4 days after stroke or TIA is associated with fewer ischaemic lesions at 1 month in AF patients. There is no increased rate of haemorrhage with early anticoagulation. These results suggest that early anticoagulation after mild-to-moderate acute ischaemic stroke associated with AF might be safe, but randomised controlled studies are needed to inform clinical practice.

Survival of beneficial microbes in liquid bioformulation and optimization of different carrier materials using RSM technique.

Soil salinity in rice cultivation areas is considered a severely limiting factor that adversely affects the quantity and quality of rice production in wetlands. Recently, the alternative use of salt-tolerant plant growth-promoting rhizobacteria (PGPR) inhabiting extreme saline conditions has gained remarkable attention and had positive effects on soil and crops. Therefore, a study has been initiated to develop a liquid biofertilizer formulation from locally isolated multi-strain salt-tolerant PGPR strains such as Bacillus tequilensis and Bacillus aryabhattai, using glycerol (5 mM), trehalose (10 mM), and polyvinylpyrrolidone (PVP) at 1% as additives to prolong the shelf-life of the bacteria. After 3 months of incubation, the bacterial population in the trehalose-supplemented mixed strain was highest at 9.73×107 CFU/mL, followed by UPMRE6 and UPMRB9 at 9.40×107 CFU/mL and 8.50×107 CFU/mL respectively. The results showed that the optimal trehalose concentration successfully prolonged the shelf-life of bacteria with minimal cell loss. Validation of quadratic optimization by response surface methodology revealed that the cell density of the mixed strain was 4.278×107 log CFU/mL after 24 h. The precision ratio was 99.7% higher than the predicted value in the minimized medium formulation: 0.267 g/mL trehalose, 1% glycerol, at 120 rpm agitation using the data analysis tools of Design Expert software. The population study confirmed the better and longer survival of salt-tolerant PGPR fortified with 10 mM trehalose, which was considered the best liquid biofertilizer formulation. Moreover, the optimized trehalose-glycerol liquid formulation can be used commercially as it is cost-effective.

Rapidly accelerated development of neutralizing COVID-19 antibodies by reducing cell line and CMC development timelines.

Since the Coronavirus Disease 2019 (COVID-19) outbreak, unconventional cell line development (CLD) strategies have been taken to enable development of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-neutralizing antibodies at expedited speed. We previously reported a novel chemistry, manufacturing, and control (CMC) workflow and demonstrated a much-shortened timeline of 3-6 months from DNA to investigational new drug (IND) application. Hereafter, we have incorporated this CMC strategy for many SARS-CoV-2-neutralizing antibody programs at WuXi Biologics. In this paper, we summarize the accelerated development of a total of seven antibody programs, some of which have received emergency use authorization  approval in less than 2 years. Stable pools generated under good manufacturing practice (GMP) conditions consistently exhibited similar productivity and product quality at different scales and batches, enabling rapid initiation of phase I clinical trials. Clones with comparable product quality as parental pools were subsequently screened and selected for late-stage development and manufacturing. Moreover, a preliminary stability study plan was devised to greatly reduce the time required for final clone determination and next-generation sequencing-based viral testing was implemented to support rapid conditional release of the master cell bank for GMP production. The successful execution of these COVID-19 programs relies on our robust, fit for purpose, and continuously improving CLD platform. The speed achieved for pandemic-related biologics development may innovate typical biologics development timelines and become a new standard in the industry.

Okra Growth, Yield and Rhizosphere Microbiome Responses to the Encapsulated Bioinoculant Application under Reduced Fertilization Regime.

There is limited evidence that Enterobacter hormaechei can improve plant physiology and yield through soil phosphate (P) and potassium (K) amelioration. This study unraveled the effect of different soil inoculation methods i.e., free-cell and encapsulated (alginate bead containing sugar-protein hydrolysate and molasses) E. hormaechei 40a with different rates of PK-fertilization on okra P and K uptake, and soil rhizosphere bacterial community. The results revealed that 3HB (half-dose PK-fertilizer + encapsulated strain 40a) had the highest soil available P (SAP) and K (SAK), as well as P and K uptake for all plant organs, followed by 3F (full-dose PK-fertilizer), 3HI (half-dose PK-fertilizer + free-cell strain 40a), and 3H (half-dose PK-fertilizer), and improved yield by up to 75.6%. Both inoculated and full-dose fertilizer treatments produced larger pods (>15 cm) compared to 3H. We discovered increased bacterial richness and diversity in both 3HB and 3HI samples compared to uninoculated treatments. Both 3HB and 3F treatments were positively correlated with the increasing abundance of Acidobacteriales, Burkholderia caballeronia paraburkholderia, Gemmataceae, and Sphingomonas along with the SAP and SAK. The plant-beneficial effect of one-time 3HB treatment on okra growth and yield was comparable to biweekly inoculation in 3HI, suggesting a new cost-effective farming approach in precision agriculture.

Gamma rays induced genetic variability in tomato (Solanum lycopersicum L.) germplasm.

The present study assessed the effectiveness of gamma radiation in inducing favorable genetic variability in tomato (Solanum lycopersicum L.). An experiment was conducted in a randomized complete block design to produce M1 generation. Significant differences were observed among the genotypes as well as between the treatments at individual plant level based on observed traits (seed germination percentage, seedling survival, plant height, number of flower clusters plant-1, number of flowers and fruits plant-1). All observed characters in the mutagenized population were adversely affected with increasing radiation dose. Results identified 450 Gy as the most damaging radiation dose followed by 300 Gy and 150 Gy. Moreover, 300 Gy treatment was identified as lethal dose (LD50) as it caused a 50% germination inhibition in almost all the evaluated genotypes. The 150 Gy treatment showed the least damaging impact and induced maximum genetic variability in almost all the genotypes under study. Character association studies were also conducted which could be utilized in the selection of desirable mutants. Correlation studies revealed an altered association among the observed parameters from positive to negative direction in 300 Gy and 450 Gy treatments as compared to control. These deviations in correlation coefficients proved that mutagenesis can break the linkage among specific loci. Furthermore, path coefficient analysis identified the growth attributes with an effective direct and indirect contribution in yield.

Sugar-Assisted Cryopreservation of Stem Cell-Laden Gellan Gum-Collagen Interpenetrating Network Hydrogels.

Tissue engineering involves the transplantation of stem cell-laden hydrogels as synthetic constructs to replace damaged tissues. However, their time-consuming fabrication procedures are hurdles to widespread application in clinics. Fortunately, similar to cell banking, synthetic tissues could be cryopreserved for subsequent central distribution. Here, we report the use of trehalose and gellan gum as biomacromolecules to form a cryopreservable yet directly implantable hydrogel system for adipose-derived stem cell (ADSC) delivery. Through a modified cell encapsulation method and a preincubation step, adequate cryoprotection was afforded at 0.75 M trehalose to the encapsulated ADSCs. At this concentration, trehalose demonstrated lower propensity to induce apoptosis than 10% DMSO, the current gold standard cryoprotectant. Moreover, when cultured along with trehalose after thawing, the encapsulated ADSCs retained their stem cell-like phenotype and osteogenic differentiation capacity. Taken together, this study demonstrates the feasibility of an "off-the-shelf" biomacromolecule-based synthetic tissue to be applied in widespread tissue engineering applications.

The impact of newly synthesized sulfonamides on soil microbial population and respiration in rhizospheric soil of wheat (Triticum aestivum L.).

Antibiotics released into agricultural fields through the manure of grazing animals could exert harmful impacts on soil microbes and plants. Antibiotics exert high impacts on environment than other pharmaceuticals due to their higher biological activity. However, little is known about their impacts on plants, despite indications that antibiotics exert negative effects on soil microorganisms, which ultimately harm the plants. It has been demonstrated that beneficial microorganisms promote plant growth and development under various stresses. This study evaluated the toxicity of four newly derived sulfonamides (SAs), i.e., 2-(phenylsulfonyl) hydrazine carbothioamide (TSBS-1), N, 2-bis phenyl hydrazine carbothioamide (TSBS-2), aminocarbonyl benzene sulfonamide (UBS-1), and N, N'-carbonyl dibenzene sulfonamide (UBS-2) on bacterial growth and soil microbial respiration. Each SA was tested at four different concentrations (i.e., 2.25, 2.5, 3, 4 mg/ml) against five rhizospheric bacterial strains, including AC (Actinobacteria sp.), RS-3a (Bacillus sp.), RS-7a (Bacillus subtilis), RS-4a (Enterobacter sp.), and RS-5a (Enterobacter sp.). Antimicrobial activity was checked by disc diffusion method, which showed that inhibition zone increased with increasing concentration of SAs. The UBS-1 resulted in the highest inhibition zone (11.47 ± 0.90 mm) against RS-4a with the highest concentration (4 mg/ml). Except TSBS-1, all sulfonamide derivatives reduced CO2 respiration rates in soil. Soil respiration values significantly increased till 6th day; however, exposure of sulfonamide derivatives suppressed microbial respiration after 6th day. On the 20th day, poor respiration activity was noted at 0.23, 0.2, and 0.4 (CO2 mg/g dry soil) for TSBS-1, UBS-1, and UBS-2, respectively. Our results demonstrate that sulfonamides, even in small concentrations, significantly affect soil microbial population and respiration. Soil microbial respiration changes mediated by sulfonamides were dependent on length of exposure and concentration. It is recommended that antibiotics should be carefully watched and their impact on plant growth should be tested in the future studies.

Thin layer drying kinetics and quality dynamics of persimmon (Diospyros kaki) treated with preservatives and solar dried under different temperatures.

Poor postharvest handling, microbial infestation, and high respiration rate are some the factors are responsible for poor storage life of perishable commodities. Therefore, effective preservation of these commodities is needed to lower the damages and extend shelf life. Preservation is regarded as the action taken to maintain desired properties of a perishable commodity as long as possible. Persimmon (Diospyros kaki) is perishable fruit with high nutritive value; however, has very short shelf-life. Therefore, effective preservation and drying is needed to extend its storage life. Drying temperature and preservatives significantly influence the quality of perishable vegetables and fruits during drying. The current study investigated the effect of different temperatures and preservatives on drying kinetics and organoleptic quality attributes of persimmon. Persimmon fruits were treated with preservatives (25% honey, 25% aloe vera, 2% sodium benzoate, 1% potassium metabisulfite, and 2% citric acid solutions) under different drying temperatures (40, 45, and 50°C). All observed parameters were significantly affected by individual effects of temperatures and preservatives, except ash contents. Similarly, interactive effects were significant for all parameters except total soluble sugars, ash contents, and vitamin C. Generally, fruits treated with citric acid and dried under 50°C had 8.2% moisture loss hour-1, 14.9 drying hours, 0.030 g H2O g-1 hr-1, 1.23° Brix of total soluble solids, 6.71 pH, 1.35% acidity, and 6.3 mg vitamin C. These values were better than the rest of the preservatives and drying temperatures used in the study. Therefore, treating fruits with citric acid and drying at 50°C was found a promising technique to extend storage life of persimmon fruits. It is recommended that persimmon fruits dried at 50°C and preserved in citric acid can be used for longer storage period.

Synchronization of Boron application methods and rates is environmentally friendly approach to improve quality attributes of Mangifera indica L. On sustainable basis.

Micronutrient deficiency in the soil is one of the major causes of mango fruit and yield's poor quality. Besides, the consumption of such a diet also causes a deficiency of micronutrients in humans. Boron deficiency adversely affects the flowering and pollen tube formation, thus decreasing mango yield and quality attributes. Soil and foliar application of B are considered a productive method to alleviate boron deficiency. A field experiment was conducted to explore the Boron most suitable method and application rate in mango under the current climatic scenario. There were nine treatments applied in three replications. The results showed that application of T8 = RD + Borax (75 g plant -1 as a basal application) + H3 BO3 (0.8% as a foliar spray) and T9 = RD + Borax (150 g plant -1 as a basal application) + H3 BO3 (0.8% as a foliar spray) significantly enhanced the nitrogen, potassium, proteins, ash, fats, fiber, and total soluble solids in mango as compared to the control. A significant decrease in sodium, total phenolics contents, antioxidant activity, and acidity as citric acid also validated the effective functioning of T8 = RD + Borax (75 g plant -1 as a basal application) + H3 BO3 (0.8% as a foliar spray) and T9 = RD + Borax (150 g plant -1 as a basal application) + H3 BO3 (0.8% as a foliar spray) as compared to control. In conclusion, T8 = RD + Borax (75 g plant -1 as a basal application) + H3 BO3 (0.8% as a foliar spray) and T9 = RD + Borax (150 g plant -1 as a basal application) + H3 BO3 (0.8% as a foliar spray) is a potent strategy to improve the quality attributes of mango under the changing climatic situation.

Field screening of diverse wheat germplasm for determining their adaptability to semi-arid climatic conditions.

Wheat (Triticum aestivum L.) is an important staple food crop for one third of global population and important crop for securing future food security. Rapid changes in the climate on global scale could be a threat for future food security. This situation urges plant breeders to explore the genetic potential of existing wheat germplasm. This study screened forty diverse wheat genotypes for their yield under two different agroclimatic conditions, i.e., Layyah and Dera Ghazi Khan, Pakistan. Data relating to plant height, peduncle length, flag leaf area, spike length, number of spikelets, number of grains per spike, thousand grain weight, chlorophyll content and grain yield were recorded. The tested wheat genotypes significantly differed for grain yield and related traits. Grain yield was positively correlated with plant height, spike length, spike number, flag leaf length, number of grains per spike, and 1000-grain weight. Biplot obtained from the cluster analysis by Euclidean method grouped the studied genotypes in 3 different groups. The genotypes exhibited 10.77% variability within quadrants, whereas 72.36% variability was recorded between quadrants according to clustering. Dendrogram grouped the tested genotypes into two main clusters. The main cluster "I" comprised of 2 genotypes, i.e., 'Seher-2006' and 'AS-2002'. The cluster "II" contained 38 genotypes based on Euclidian values. Genotypes within same cluster had smaller D2 values compared to those belonging to other clusters. The genetic relationships of genotypes provide useful information for breeding programs. Overall, the results revealed that genotypes 'Line 9733', 'Bhakar-2002', 'Line A9' and 'SYN-46' had better yield and yield stability under climatic conditions of southern Punjab. Therefore, these genotypes could be recommended for general cultivation in the study region.

The impact of PEG-induced drought stress on seed germination and seedling growth of different bread wheat (Triticum aestivum L.) genotypes.

Wheat is an important crop, used as staple food in numerous countries around the world. However, wheat productivity is low in the developing world due to several biotic and abiotic stresses, particularly drought stress. Non-availability of drought-tolerant wheat genotypes at different growth stages is the major constraint in improving wheat productivity in the developing world. Therefore, screening/developing drought-tolerant genotypes at different growth stages could improve the productivity of wheat. This study assessed seed germination and seedling growth of eight wheat genotypes under polyethylene glycol (PEG)-induced stress. Two PEG-induced osmotic potentials (i.e., -0.6 and -1.2 MPa) were included in the study along with control (0 MPa). Wheat genotypes included in the study were 'KLR-16', 'B6', 'J10', '716', 'A12', 'Seher', 'KTDH-16', and 'J4'. Data relating to seed germination percentage, root and shoot length, fresh and dry weight of roots and shoot, root/shoot length ratio and chlorophyll content were recorded. The studied parameters were significantly altered by individual and interactive effects of genotypes and PEG-induced osmotic potentials. Seed germination and growth parameters were reduced by osmotic potentials; however, huge differences were noted among genotypes. A reduction of 32.83 to 53.50% was recorded in seed germination, 24.611 to 47.75% in root length, 37.83 to 53.72% in shoot length, and 53.35 to 65.16% in root fresh weight. The genotypes, 'J4', 'KLR-16' and 'KTDH-16', particularly 'J4' better tolerated increasing osmotic potentials compared to the rest of the genotypes included in the study. Principal component analysis segregated these genotypes from the rest of the genotypes included in the study indicated that these can be used in the future studies to improve the drought tolerance of wheat crop. The genotype 'J4' can be used as a breeding material to develop drought resistant wheat genotypes.

Evaluation of seedling age and nutrient sources on phenology, yield and agrometeorological indices for sweet corn (Zea mays saccharata L.).

The field experiment was conducted during Kharif season of 2020 at Agronomy farm of Faculty of Agriculture, Wadura, SKUAST-K to study the influence of age of seedling and sources of nutrients on phenology, yield and agrometeorological indices for sweet corn. The experiment included two factors viz. age of seedlings (12, 22 and 32 days old seedling) and sources of nutrients (control, RDF, 50 percent RDF + FYM @ 12 t ha-1, 50 percent RDF + vermi-compost @ 4 t ha-1 and 50 percent RDF + poultry manure @ 2 t ha-1) tested in RCBD with three replications. Transplanting 12 days old seedlings required maximum number of days to attain different phenological stages, thereby accumulated maximum heat units followed by 22 days old seedlings. While as transplanting 22 days old seedling recorded significantly highest HUE, HTUE, PTUE and HyTUE and consequently resulted in the highest green cob and biological yield compared to other ages of seedlings. Among various sources of nutrients, application of 50 per cent RDF + poultry manure @ 2 t ha-1 took maximum number of days to attain various phenophases thereby accumulated maximum heat units and registered highest HUE, HTUE, PTUE and HyTUE followed by application of 100 per cent RDF.

Assessment of the bioaccumulation pattern of Pb, Cd, Cr and Hg in edible fishes of East kolkata Wetlands, India.

Waste water fed pisciculture is nowadays a common feature in aquaculture belts across the globe. East Kolkata Wetlands (EKW) a nature's wonder where waste water fed natural aquaculture beltis is active for more than 70 years now and is efficiently operating as a natural waste management system. The peri urban wetland is also a site of international importance and is listed in Ramsar. Field and lab-based investigations were carried out using three commonly edible carp variety of fishes such as Rohu (Labeorohita), Catla (Catlacatla) and Nile Tilapia (Oreochromisniloticus) collected from ponds (bheries) of the wetland located on the eastern fringes of Kolkata, India. The lab-based analysis revealed the presence of toxic metals such as Cr, Pb, Cd and Hg in the samples with the seasonal order of accumulation being monsoon > post-monsoon > winter > pre-monsoon in the successive years of 2016, 2017 and 2018. Bio-accumulation of toxic heavy metals in fishes follows the order Tilapia > Rohu > Catla where as the bioaccumulation pattern of toxic metals shows the trend Pb > Cd > Cr > Hg across all the seasons and years. The ambient media was also investigated to understand in detail the bioaccumulation pattern at different trophic levels of the ecosystem. Water and sediments were analyzed to evaluate the contamination of toxic heavy metals from point and non-point sources. Current study shows the observed bioaccumulation pattern of the toxic heavy metals in one of the fragile ecosystems that raises an important question of environmental safety in the food we intake on daily basis.

Sustainable phosphorous management in two different soil series of Pakistan by evaluating dynamics of phosphatic fertilizer source.

Phosphorous (P) plays the prominent role to promote the plants storage functions and structural roles, as it is recognized as a vital component of ADP, ATP, Cell wall as well as a part of DNA. Soils acts as the sink to supply P to plants because soil pH and its physical condition are the main factor which regulate the solubility and availability P element. Phosphorus is not deficient in Pakistani soils but its availability to plants is the serious matter of concern. A pot experiment was conducted to evaluate P dynamics in two different soil series of Pakistan (Bahawalpur and Lyallpur) using Maize as test crop. The treatments applied were T0: Control (without any fertilizer), T1: Recommended DAP @648 mg pot-1, T2: Half dose DAP @324 mg pot-1, T3: Recommended rate of TSP @900 mg pot-1, T4: Half dose TSP @450 mg pot-1. Soil analysis showed that Bahawalpur soil has sandy clay loam texture with 33% clay and Lyallpur series has sandy loam texture with 15.5% clay; furthermore, these soil contain 4.6 and 2.12% CaCO3 respectively. Results showed an increase in P concentration in roots (23 mg kg-1) with the application of half dose of TSP in Lyallpur series and lowest in Bahawalpur series (14.6 mg kg-1) at recommended dose of DAP. Concentration of P in shoots responded the same; increase at half dose of TSP (16.7 mg kg-1) and lowest at full dose of DAP in Bahawalpur series as (15.58 mg kg-1). Adsorbed P (17 mg kg-1) was recorded highest in Bahawalpur soil with more clay amount in pot with DAP application but lower in Lyallpur soil series (14 mg kg-1) with the application of applied TSP. The PUE was recorded highest in Lyallpur series with the application of half dose of TSP and it was 61% more than control and was Highest in Bahawalpur series was with the application of recommended dose of DAP is 72% more than control treatment. On estimation; results showed that applied sources made an increase in P availability than control, but TSP gave better P uptake than DAP unless of rates applied. Soil of Lyallpur series showed better uptake of P and response to applied fertilizers than Bahawalpur series which showed more adsorption of P by high clay and CaCO3 amount. Conclusively, the study suggested that soil series play a crucial role in choosing fertilizer source for field application.

Bioprospecting microwave-alkaline hydrolysate cocktail of defatted soybean meal and jackfruit peel biomass as carrier additive of molasses-alginate-bead biofertilizer.

The extraction of soluble hydrolysate protein and sugar from a biomass cocktail of defatted soybean meal (DSM) and jackfruit peel (JP) was examined using microwave-alkaline hydrolysis by varying the NaOH concentrations (0.04-0.11 M) and residence times (2-11 min). Based on the central composite design, the optimized parameters were achieved at 0.084 M NaOH concentration (100 mL), for 8.7 min at 300 W microwave power level to obtain the highest protein (5.31 mg/mL) and sugar concentrations (8.07 mg/mL) with > 75% recovery. Both raw and detoxified hydrolysate (using activated carbon) were correspondingly biocompatible with Enterobacter hormaechei strain 40a (P > 0.05) resulting in maximal cell counts of > 10 log CFU/mL. The optimized hydrolysate was prepared as an additive in molasses-alginate bead encapsulation of strain 40a. Further evaluation on phosphate and potassium solubilization performance of the encapsulated strain 40a exhibited comparable results with those of free cell counterpart (P > 0.05). The DSM-JP hydrolysate cocktail holds potential as a carrier additive of encapsulated-cell bead biofertilizers in order to sustain bacterial cell quality and consequently improve crop growth and productivity.

Aerobic Methanotrophy and Co-occurrence Networks of a Tropical Rainforest and Oil Palm Plantations in Malaysia.

Oil palm (OP) plantations are gradually replacing tropical rainforest in Malaysia, one of the largest palm oil producers globally. Conversion of lands to OP plantations has been associated with compositional shifts of the microbial community, with consequences on the greenhouse gas (GHG) emissions. While the impact of the change in land use has recently been investigated for microorganisms involved in N2O emission, the response of the aerobic methanotrophs to OP agriculture remains to be determined. Here, we monitored the bacterial community composition, focusing on the aerobic methanotrophs, in OP agricultural soils since 2012, 2006, and 1993, as well as in a tropical rainforest, in 2019 and 2020. High-affinity methane uptake was confirmed, showing significantly lower rates in the OP plantations than in the tropical rainforest, but values increased with continuous OP agriculture. The bacterial, including the methanotrophic community composition, was modified with ongoing OP agriculture. The methanotrophic community composition was predominantly composed of unclassified methanotrophs, with the canonical (Methylocystis) and putative methanotrophs thought to catalyze high-affinity methane oxidation present at higher relative abundance in the oldest OP plantation. Results suggest that the methanotrophic community was relatively more stable within each site, exhibiting less temporal variations than the total bacterial community. Uncharacteristically, a 16S rRNA gene-based co-occurrence network analysis revealed a more complex and connected community in the OP agricultural soil, which may influence the resilience of the bacterial community to disturbances. Overall, we provide a first insight into the ecology and role of the aerobic methanotrophs as a methane sink in OP agricultural soils.

The impact of different plant extracts on population suppression of Helicoverpa armigera (Hub.) and tomato (Lycopersicon esculentum Mill) yield under field conditions.

Helicoverpa armigera (Hub.) is a destructive pest of the tomato (Lycopersicon esculentum Mill) crop in Pakistan. Although insecticides are the primary management strategy used to control H. armigera, most of them are not effective due to considerable toxic residual effects on the fruits. Nonetheless, H. armigera is rapidly evolving resistance against the available pesticides for its management. This situation calls upon the need of alternative management options against the pest. Different plant extracts have been suggested as a viable, environment-friendly option for plant protection with minimal side effects. Furthermore, the plant extracts could also manage the insect species evolving resistance against pesticides. This study evaluated the efficacy of different plant extracts (i.e., Neem seed, turmeric, garlic and marsh pepper) against H. armigera. Furthermore, the impact of the plant extracts on growth and yield of tomato crop was also tested under field conditions. The results revealed that all plant extracts resulted in higher mortality of H. armigera compared to control. Similarly, the highest plant height was observed for the plants treated with the plant extracts compared to untreated plants. Moreover, the highest tomato yield was observed in plants treated with plant extracts, especially with neem seed (21.013 kg/plot) followed by pepper extract (19.25 kg/plot), and garlic extract 18.4 kg/plot) compared to the untreated plants (8.9 kg/plot). It is concluded that plant extracts can be used as eco-friendly approaches for improving tomato yield and resistance management of H. armigera.

Seed germination ecology of Conyza sumatrensis populations stemming from different habitats and implications for management.

Conyza sumatrensis (Retz.) E. H. Walker is an obnoxious weed, emerging as an invasive species globally. Seed germination biology of four populations of the species stemming from arid, semi-arid, temperate, and humid regions was determined in this study. Seed germination was recorded under six different environmental cues (i.e., light/dark periods, constant and alternating day and night temperatures, pH, salinity, and osmotic potential levels) in separate experiment for each cue. Populations were main factor, whereas levels of each environmental cue were considered as sub-factor. The impact of seed burial depths on seedling emergence was inferred in a greenhouse pot experiment. Seed germination was recorded daily and four germination indices, i.e., seed germination percentage, mean germination time, time to reach 50% germination, and mean daily germination were computed. Tested populations and levels of different environmental cues had significant impact on various seed germination indices. Overall, seeds stemming from arid and semi-arid regions had higher seed germination potential under stressful and benign environmental conditions compared to temperate and humid populations. Seed of all populations required a definite light period for germination and 12 hours alternating light and dark period resulted in the highest seed germination. Seed germination of all populations occurred under 5-30°C constant and all tested alternate day and night temperatures. However, the highest seed germination was recorded under 20°C. Seeds of arid and semi-arid populations exhibited higher germination under increased temperature, salinity and osmotic potential levels indicating that maternal environment strongly affected germination traits of the tested populations. The highest seed germination of the tested populations was noted under neutral pH, while higher and lower pH than neutral had negative impact on seed germination. Arid and semi-arid populations exhibited higher seed germination under increased pH compared to temperate and humid populations. Seed burial depth had a significant effect on the seedling emergence of all tested populations. An initial increase was noted in seedling emergence percentage with increasing soil depth. However, a steep decline was recorded after 2 cm seed burial depth. These results indicate that maternal environment strongly mediates germination traits of different populations. Lower emergence from >4 cm seed burial depth warrants that deep burial of seeds and subsequent zero or minimum soil disturbance could aid the management of the species in agricultural habitats. However, management strategies should be developed for other habitats to halt the spread of the species.