High Yield Research Articles

Synergistic Nanoscale Mn3O4-CoO-Co Heterojunctions for Boosting the Selectivity in Hydrogenation of Nitrostyrenes and Nitroarenes.

Metal-metal oxide interface catalysts are in high demand for advanced catalytic applications due to their multi-component active sites, which facilitate synergistic cooperation where a single component alone cannot effectively promote the desired reaction. Demonstrated herein graphene oxide-supported nanoscale Mn3O4-CoO-Co as highly efficient catalysts for hydrogenation of nitro styrenes/nitro arenes to amino styrenes/arenes under mild reaction conditions (0.5 MPa and 100 °C in 1:1 THF/water). Charge relocalization at the Co-CoO-Mn3O4 heterojunction interfaces, primarily driven by Mn3O4, significantly improves reaction selectivity. Replacing Mn3O4 with MnO or using other supported bimetallic CoMnOx catalysts decreases selectivity, leading to the formation of a mixture of products. The catalyst demonstrated remarkable selectivity in converting nitro groups to amines, even in the presence of highly reactive functional groups such as C=C, O-C=O, C=O, C≡N, chalcones, and halides. It also exhibited high yields, multiple reusability, and a broad substrate scope. This study demonstrates how Mn3O4, in synergy with CoO-Co, fine-tunes selectivity, paving the way for the development of advanced metal-metal oxide interface catalysts to enhance both selectivity and efficiency in organic transformations.

Evaluation of Early Maturing Soya Bean (Glycine max.) Genotypes for Pod Shattering Resistance and High Yielder at Western Tigray, Ethiopia

Pod shattering is occurring after the pods reached maturity which can cause considerable yield loss in soya bean crop. The aims of this study were to evaluate the pod shattering resistance and the performance of the agronomic traits of several soybean genotypes. Field experiment was conducted at Humera district during 2018 cropping season using sixteen soya bean genotypes. Treatments were evaluated with RCBD and replicated three times. Pod shattering, number of branches, seed per pod, pod per plant, hundred seed weight and seed yield was shows a significant (P<0.01) variation among genotypes. While; there was not a significant variation among genotypes in terms of the number of days from planting to flowering and maturity. The mean pod shattering was ranged from scale1.25 to 4. Maximum amount of pod shattering was measured in the genotypes PI423959 (4), PI506764 (3.75) and PI417085 (3.25). However, minimum amount of pod shattering was measured in the genotypes of PI417116 (1.25), Desha (1.75) and PI417132 (2). Highest yield was obtained from the genotypes PI714116 (19.29kg/ha) and PI417132 (16.54kg/ha). However, lowest yield was obtained from the genotypes Williams (check) (6.7kg/ha) followed by PI223963 (8.53 kg/ha) and Nova (check) (9.86 kg/ha).

Asymmetric Synthesis of Benzofuranones with a C3 Quaternary Center via an Addition/Cyclization Cascade Using Noncovalent N-Heterocyclic Carbene Catalysis.

An asymmetric addition/cyclization cascade of amidoesters and iminoquinones is developed using noncovalent N-heterocyclic carbene (NHC) catalysis. The process enables access to various functionalized benzofuranones with an all-carbon quaternary stereocenter with high yields and ee values. The reaction displays a broad substrate scope. Via product modifications, enantioenriched synthesis of biologically relevant spirocyclic lactones and lactams is achieved. Substrate activation via noncovalent interaction with NHC is suggested for the process.

Identification of a Novel Transasparaginase Activity of Bacillus subtilis (bTG) for Sequence-Specific Bioconjugation.

The ability of Bacillus subtilis transglutaminase (bTG) to functionalize BSA has been investigated using peptide mapping experiments. Interestingly, the conjugation was not detected on a glutamine but on an asparagine residue. A sequence determination study was further performed, and a sequence of 10 amino acids for site-specific conjugation was identified. A monobody showing no native reactivity with the bTG enzyme was produced with the identified peptide sequences and successfully conjugated to various types of substrates in very high yields (>90%) with a 1/1/1.5 ratio of protein/amine/enzyme. Direct conjugation to the amino linker of a small interfering RNA (siRNA) was achieved in good yield, and no impact on the siRNA activity was observed following the conjugation. The identified sequences were further engineered in VHH and IgG scaffolds, and successful conjugation could also be observed with both small molecules and siRNA, confirming the potential of bTG for site-specific enzymatic bioconjugation.

Diastereoselective Stereo-Divergent Synthesis of Spiroindolines via Ligand-Controlled Silver(I)-Catalyzed Asymmetric [3 + 2] cycloadditions

A series of spirocyclic pyrrolidine derivatives with quaternary stereocenters was constructed via asymmetric [3 + 2] cycloaddition reactions between imino esters and 2-oxindole. By ingenious use of ligands on the silver catalyst allows stereoselectivity to either endo-isomers (ee up to 98 %, yields up to 96 %) with (R, Rp)-Ph-Phosferrox or exo-isomers (ee up to 99 %, yields up to 87 %) with (R)-BINAP. This work is notable for its high yields, broad substrate adaptability and excellent enantioselectivity.

Dearomative Alkylation-Based Two-Step cis-Diastereoselective Synthesis of Indoline-2,3-Fused Chromans and Tetrahydropyrans.

Herein, we describe a two-step, cis-diastereoselective synthesis of indoline-2,3-fused chromans from 3-substituted indoles. The method proceeds without intermediacy of ortho-quinone methides and leverages the dual function of TBS-protected 2-hydroxybenzyl iodides both as highly reactive alkylating agents in a t-BuONa/Et3B-promoted dearomative alkylation step and as a source of masked phenoxide nucleophiles in a subsequent TBAF-induced one-pot deprotection-cyclization step of the resulting indolenines. Importantly, this two-step protocol can also be extended to access indoline-2,3-fused tetrahydropyrans. These syntheses of indoline-2,3-fused chromans and tetrahydropyrans proceed with operational convenience, use easily accessible substrates and reagents, and feature broad substrate scope, high yields and complete diastereoselectivity. Furthermore, the synthesized products have the potential to undergo late-stage functionalization.

Integrating environmental remediation with biodiesel production from toxic non-edible oil seeds (Croton bonplandianus) using a sustainable phyto-nano catalyst

In the current situation of the environmental uprising toxicology, rising global temperature, and energy-depleting urges to explore and discover more renewable and greener ecological-benefiting energy resources. Biobased renewable fuels generated by using waste products can help in waste management, climate change mitigation, and a low-carbon future. The main objective of this research is to produce environment-friendly and cost-effective biofuel. The potentiality of the novel, toxic, waste, and inedible feedstock Croton bonplandianus was evaluated for biodiesel synthesis through transesterification utilizing a Phyto-nano catalyst of potassium oxide prepared by Croton bonplandianus floral stalk's aqueous extract focusing on waste management. Phyto-nano catalyst characterization was done through innovative tools such as Scanning Electron Microscopy (SEM), Energy Dispersive Spectroscopy (EDS), Zeta Potential (ZP), X-Ray Diffraction (XRD), Fourier Transformed Infrared spectroscopy (FTIR), and Diffuse Reflectance Spectroscopy (DRS). The characterization results revealed that the potassium oxide phyto-nanocatalyst possesses an average nanoparticle size of 44.5 nm. This size is optimal for enhanced catalytic activity, indicating significant potential for efficient catalysis. The highest yield (94 %) of biodiesel was secured at optimized reaction conditions of catalyst quantity (0.50 wt%), reaction time (180 min), methanol: oil ratio (9:1), and reaction thermal point (70 °C). Transformation of triglycerides to methyl esters was confirmed by GC/MS, NMR, and FTIR techniques. A total of 21 methyl esters were observed in Croton bonplandianus biodiesel confirmed via GC/MS results. Evaluation of fuel properties was done and matched with international fuel standards. The conclusive remarks for the conducted research are that Croton bonplandianus has a high potential for biodiesel production by applying Phyto-nanocatalysts of potassium oxide while dealing with hazardous environmental conditions and waste management. Phyto nanocatalyst of potassium oxide can be reused and gives the same yield after several cycles of reusability, this reusability of heterogenous Phyto nanocatalyst can reduce to total cost of biodiesel production and can contribute towards circular economy.

Enhanced Fluorescence in Green ZnSeTe Quantum Dots Using Gradient Layer Technique

AbstractA novel method is presented for synthesizing green ZnSeTe quantum dots (QDs) that demonstrate high photoluminescence quantum yield (PLQY) and a narrow full width at half maximum (FWHM). In this synthesis, a gradient layer is created between the ZnSeTe core and the ZnSe inner shell by utilizing the different reactivities of precursors. This innovative approach aims to reduce lattice mismatch and minimize interfacial defects, thereby enhancing the fluorescence properties of ZnSeTe QDs. Using the gradient layer, green QDs are achieved with a FWHM of 36 nm and a PLQY of 90%, surpassing previous results. Detailed structural analyses confirmed a reduction in stacking faults and strain in the QDs with a gradient layer, leading to enhanced fluorescence properties. The QDs with a gradient layer are successfully integrated into the emissive layer of a quantum dot light–emitting diode device, demonstrating superior performance compared to QDs without the gradient layer.

Activating Two-Photon Silica Nanoamplifier-Based CHA and FRET for Accurate Ratiometric Bioimaging of Intracellular MicroRNA.

In situ visualization of microRNA (miRNA) in cancer cells and diseased tissues is essential for advancing our comprehension of the onset and progression of associated diseases. Two-photon (TP) imaging, as an imaging technology with high spatiotemporal resolution, deep tissue penetration, and accurate target quantification, has distinctive advantages over single-photon imaging and has attracted increasing attention. Extensive research has been conducted on two-photon dye-doped silica nanoparticles, which exhibit a large two-photon absorption (TPA) cross-section, high fluorescence quantum yield, and excellent biocompatibility. However, the low abundance of RNA in tumor cells leads to insufficient signal output. Based on functional nucleic acid, a catalyzed hairpin self-assembly (CHA) signal amplification strategy, which has simplicity, robustness, and nonenzymatic characteristics, can achieve the amplification of DNA or RNA signals. Here, a two-photon silica nanoamplifier (TP-SNA) utilizing TP dye-doped silica nanoparticles (SiNPs) and functional nucleic acid was constructed, employing triggering catalyzed hairpin self-assembly and fluorescence resonance energy transfer (FRET) for highly sensitive detection and precise TP imaging of endogenous miRNAs in tumor cells and tissues at varying depths. The TP-SNA demonstrated the capability to detect miR-203 with a detection limit of 33 pM. The maximum two-photon tissue penetration depth of the two-photon nanoamplifier was 210 μm. The two-photon nanoamplifier developed in this study makes full use of the advantages of accurate TP ratiometric bioimaging and the CHA signal amplification strategy, which shows good application value for future transformation into clinical diagnosis.

Design and synthesis of surface-decorated zinc-doped carbon quantum dots as fluorescent probes for tartrazine detection in real food samples exploiting the inner filter effect mechanism

We developed a rapid and sensitive method for detecting tartrazine (Tar), a common food colorant, using zinc-doped carbon quantum dots (Zn-doped CQDs). Synthesized from biowaste pigeon pod shells and zinc acetate, the Zn-doped CQDs exhibited a high quantum yield (47.63%) and were characterized by photo-stability, non-toxicity, and water solubility. The inner filter effect (IFE) allows selective attenuation of Zn-doped CQD fluorescence by Tar, enabling precise detection. The approach obtains a low detection limit of 20.35 ng/mL and a quantification limit of 61.69 ng/mL within a linear range of 0–100 ng/mL under optimal conditions when employing the Box-Behnken statistical design. The approach demonstrates excellent reproducibility, successfully detecting Tar in real food samples with recovery rates ranging from 89% to 102.2%. This high accuracy confirms the sensor's reliability and effectiveness in practical applications. This cost-effective and rapid technique presents a promising solution for monitoring Tar levels in food products.

In-situ electrochemical reconstruction and modulation of adsorbed hydrogen coverage in cobalt/ruthenium-based catalyst boost electroreduction of nitrate to ammonia

The electroreduction of nitrate offers a promising, sustainable, and decentralized route to generate valuable ammonia. However, a key challenge in the nitrate reduction reaction is the energy efficiency of the reaction, which requires both a high ammonia yield rate and a high Faradaic efficiency of ammonia at a low working potential (≥−0.2 V versus reversible hydrogen electrode). We propose a bimetallic Co–B/Ru12 electrocatalyst which utilizes complementary effects of Co–B and Ru to modulate the quantity of adsorbed hydrogen and to favor the specific hydrogenation for initiating nitrate reduction reaction at a low overpotential. This effect enables the catalyst to achieve a Faradaic efficiency for ammonia of 90.4 ± 9.2% and a remarkable half-cell energy efficiency of 40.9 ± 4% at 0 V versus reversible hydrogen electrode. The in-situ electrochemical reconstruction of the catalyst contributes to boosting the ammonia yield rate to a high level of 15.0 ± 0.7 mg h−1 cm−2 at −0.2 V versus reversible hydrogen electrode. More importantly, by employing single-entity electrochemistry coupled with identical location transmission electron microscopy, we gain systematic insights into the correlation between the increase in the catalyst’s active sites and its structural transformations during the nitrate reduction reaction.

AMMI and GGE Biplot Analysis for Selection of Some High Yielding Terminal Heat Stress Tolerant Wheat (Triticum aestivum) Genotypes in Bangladesh

AMMI and GGE Biplot Analysis for Selection of Some High Yielding Terminal Heat Stress Tolerant Wheat (Triticum aestivum) Genotypes in Bangladesh

Transcriptomic and metabolomic analysis of poplar response to feeding by Hyphantria cunea

Populus cathayana × canadansis ‘Xinlin 1’ (‘P.‘xin lin 1’) with the characteristics of rapid growth and high yield, is frequently attacked by herbivorous insects. However, little is known about how it defenses against Hyphantria cunea (H. cunea) at molecular and biochemical levels. Differences in the transcriptome and metabolome were analyzed after ‘P. ‘xin lin 1’ leaves were fed to H. cunea for 0h, 2h, 4h, 8h, 16h and 24h. In the five comparison groups including 2h vs. CK, 4h vs. CK, 8h vs. CK, 16h vs. CK, and 24h vs. CK, a total of 8925 genes and 842 metabolites were differentially expressed. A total of 825 transcription factors (TFs) were identified, which encoded 56 TF families. The results showed that the top four families with the highest number of TFs were AP2/ERF, MYB, C2C2, bHLH. Analyses of leaves which were fed to H. cunea showed that the differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) were significantly enriched in plant hormone signal transduction pathway, MAPK signaling pathway, flavonoid, flavone and flavonol and anthocyanin biosynthesis pathway. Additionally, there were a number of genes significantly up-regulated in MAPK signaling pathway. Some compounds involved in plant hormone signal transduction and flavonoid/flavone and flavonol/ anthocyanin pathways such as jasmonic acid (JA), jasmonoyl-L-Isoleucine (JA-Ile), kaempferol and cyanidin-3-O-glucoside were induced in infested ‘P.‘xin lin 1’. This study provides a new understanding for exploring the dynamic response mechanism of poplar to the infestation of H. cunea.

B-174 Quantification of vitamin C in serum as a biomarker using the Laser Diode Thermal Desorption technique “LDTD”/Mass Spectrometry (Luxon)

Abstract Background Ascorbic acid (AA) or vitamin C, is a crucial substance that is involved in numerous physiological processes in living beings, including collagen production and antioxidant activity. When ascorbic acid is exposed to moisture, air, heat, light and oxygen, it oxidizes and turns into dehydroascorbic acid (DHAA). Our objective is to describe the sample preparation procedure and total quantification of AA and DHAA in human serum sample based on LDTD-MS/MS, and demonstrate its efficiency in quantification, combined with analysis selectivity and rapidity. Methods The Luxon is a sample introduction and ionization source that used a fiber-coupled laser diode to thermally desorb neutral molecules and charge them using a corona needle so they can be analyzed by mass spectrometry. Due to the instability of ascorbic acid, plasma samples were enriched and stabilized with an acidic solution. Sample preparation utilized a phosphate-containing buffer to minimize interferents and help volatize the analyte, followed by a precipitation step using acetonitrile to further clean the extract. Stable isotope-labeled internal standard (13C6 ascorbic acid) and homemade calibration curve and water quality controls were used for quantification. The analysis was performed using a Sciex 6500+ Triple Quad in APCI (Atmospheric Pressure Chemical Ionization) negative mode. Calibration curves ranging from 1 to 100 μg/mL and QCs were prepared in ascorbic acid-depleted serum (kept at room temperature/exposed to light for 4 days). The peak area against the internal standard (IS) ratio was used to normalize the signal. The calibration curves were plotted using the peak area ratio and the nominal concentration of standards. The following parameters were assessed for quantitative analysis: linearity, precision, accuracy, carryover and parallelism. A linear regression model with 1/X2 weighting was used to obtain the calibration curve equation and determine the correlation coefficient (r2). Parallelism was assessed by comparing the correlation coefficient of calibration curves prepared in water to six curves prepared with biological matrixes. Intra and inter assay precision and accuracy were determined by measuring QC samples ranging from low to high concentration in five replicates for 3 consecutive days. Results Calibration curves (n=3) showed excellent linearity. The calibration curve resulted in a linear response throughout the concentration range, the correlation coefficient was 0.99. For each quality control concentrations CVs were <10% complying with desirable specifications for imprecision and accuracy. Intra-assay CVs were 3.0-8.89% and inter-assay CVs were 7.64-9.79% for precision which reflects the agreement between repeated measurements. The results for accuracy showed similar results, intra-assay CVs were 3.83-9.20% and inter-assay CVs were 5.0-9.7%. Conclusions The new LDTD-MS/MS method described here has been validated as an effective technique to reliably and rapidly quantify ascorbic acid. Simple sample preparation, rapid acid stabilization combined with high yield allow analysis without degradation responses.

Breeding Maize Hybrids with Improved Drought Tolerance Using Genetic Transformation

Drought is considered the main agricultural menace, limiting the successful realization of land potential, and thereby reducing crop productivity worldwide. Therefore, breeding maize hybrids with improved drought tolerance via genetic manipulation is necessary. Herein, the multiple bud clumps of elite inbred maize lines, DH4866, Qi319, Y478 and DH9938, widely used in China, were transformed with the Escherichia coli betA gene encoding choline dehydrogenase (EC 1.1.99.1), a key enzyme in the biosynthesis of glycine betaine from choline, using Agrobacterium to generate betA transgenic lines. After 3–4 consecutive generations of self-pollination in these transgenic plants, progenies with a uniform appearance, excellent drought tolerance, and useful agricultural traits were obtained. We evaluated the drought tolerance of T4 progenies derived from these transgenic plants in the field under reduced irrigation. We found that a few lines exhibited much higher drought tolerance than the non-transformed control plants. Transgenic plants accumulated higher levels of glycine betaine and were relatively more tolerant to drought stress than the controls at both the germination and early seedling stages. The grain yield of the transgenic plants was significantly higher than that of the control plants after drought treatment. Drought-tolerant inbred lines were mated and crossed to create hybrids, and the drought tolerance of these transgenic hybrids was found to be enhanced under field conditions compared with those of the non-transgenic (control) plants and two other commercial hybrids in China. High yield and drought tolerance were achieved concurrently. These transgenic inbred lines and hybrids were useful in marginal and submarginal lands in semiarid and arid regions. The betA transgene can improve the viability of crops grown in soils with sufficient or insufficient water.

Effect of Pinching on Growth, Yield, and Quality of African Marigold (Tagetes erecta L.) cv. Pusa Narangi Gainda

Marigold is a very common and a hardy flower crop cultivated throughout the world. Pinching is an important intercultural practice commonly followed in cultivation of marigold however, this needs to be standardized for getting higher yields in commercial cultivation. Hence, the present investigation was conducted at Rajasthan Agricultural Research Institute, Durgapura, Jaipur, Division of Horticulture, during 2020-21 and 2021-22 to standardize the commercial pinching practice for getting higher productivity. Three levels of pinching (no pinching, single at 20 days and double at 20 and 40 days after transplanting) were used in the course of the research arranged using a factorial randomized block layout with three replications applied to African marigold variety Pusa Narangi Gainda. The results revealed that treatment P0 (No pinching) was discovered to be the finest in increment in height of plant, early flower bud initiation flower opening and treatment P2 (Double pinching) in boosting spread of plant, primary branches, length of leaf, leaf width, leaf area, plant fresh weight, plant dry weight, total biomass, root- shoot ratio, 50% flowering, flower diameter, stalk and ray floret length, fresh and dry flower weight, mass and number of flower per plant, flower yield additionally improved quality characteristics like chlorophyll and xanthophyll content in marigold. Hence, it might be concluded that application of double pinching can be helpful and recommended in improving yield as well as quality characteristics in commercial cultivation of marigold.

B-185 A Comparison Between Two Oral Sample Collection Devices on Preserving High-Quality Genomic DNA

Abstract Background High-quality genomic testing significantly enhances the knowledge on gene-disease association, and greatly benefits future biomedical research and precision medicine. However, widely used saliva sample collection presents challenges with high-quality and quantity of human DNA and high bacterial DNA ratio. In this study, we validated the performance of two commercially available oral collection devices for their ability to preserve high-quality genomic DNA long-term at room temperature. Methods Fifty-one buccal samples in total from twelve donors were collected using two kinds of commercially available oral sample collection devices (iSWAB®-DNA (Device 1) and OCD-100 (Device 2)). All of the collected samples were stored at ambient temperature for up to 1 year. Collected samples were extracted with QIAamp Blood Mini Kit (Qiagen) at baseline, 30 days, 60 days, and 1 year. All DNA yields were measured using a Nanodrop One Microvolume UV-Vis Spectrophotometer; TapeStation System was used for determining the integrity of DNA. The bacterial DNA ratio was estimated by quantitative PCR with primers targeting the 16S rRNA gene known to be present in prokaryotes but not eukaryotes. Results Device 1 had higher yield (Device 1: 18.59 ± 8.98; Device 2: 6.44 ± 4.62), lower bacterial DNA ratio (Device 1: 2.06 ± 1.93; Device 2: 3.79 ± 4.58), and higher overall DNA integrity (Device 1: 6.1 ± 0.31; Device 2: 5.42 ± 0.96) compared to Device 2. Device 1 demonstrated longer post-collection sample stability at ambient temperature compared with Device 2 regarding DNA yield, bacterial DNA ratio and DNA integrity. No significant changes in DNA yield, integrity, and bacterial DNA ratio were observed after one year of storage in Device 1. No significant changes in DNA yield, integrity, and bacterial DNA ratio were observed after 60 days of storage in Device 2. However, Device 2 showed a significant increase in DNA yield, bacterial DNA ratio, and DNA integrity comparing baseline to 1 year. Conclusions Device 1 preserves higher quantity and quality of DNA from oral samples at ambient temperature for up to 1 year. These advantages significantly enhance efficiency and cost-effectiveness in downstream applications such as genotyping and sequencing, making it a valuable tool in genomics research with a commendable success rate.

Evaluation of Agronomic Performance and Resistance to Cercospora (Cercospora sojina) Leaf Spot of Selected Soya bean Genotypes in Western Tigray, Ethiopia

Aim of the current study was to evaluate soya bean genotypes for yield and yield traits and cercospora (Cercosporasojina) leaf spot disease under field condition. Field experiment was conducted at Humera district during 2018 cropping season using 16soya bean genotypes. Treatments were evaluated with RCBD and replicated three times. Number of days from planting to days of flowering and days of maturity, number of pod per plant, pod shattering, seed yield and cercospora leaf spot disease were showed a significant (P<0.01) variation among genotypes. All varieties were moderately resistance to pod shattering except PN13-1 (3.1) was moderately susceptible and Parc-3, Parc-5 and PN13-10were resistance. Highest yields were recorded from genotypes PN13-2 at 20.32q/ha and PN13-9 at 19.77q/ha. On the other hand, the lowest yields were recorded from genotypes PN13-3 at 7.49q/ha and PN13-1 at 9.95q/ha. Genotypes PN13-2 and PN13-7 have been identified as displaying resistance to Cercospora leaf spot. Highest disease severity recorded in genotypes Awassa 04 and PN13-1. Further research is needed in future to cover a wide range of environments and genotypes to screen high yielder and cercospora leaf spot disease resistance.

Yields, nutrient-use efficiencies and production economics of wheat (Triticum aestivum) as influenced by foliar nano urea

The nano fertilizers are becoming popular in crop production sector due to their higher nutrient use efficiency and also enhance the productivity and quality while requiring less input and being environmentally safe. In order to assess the effect of foliar application of nano urea on growth and yield performances of wheat (Triticum aestivum L.) and to evaluate nitrogen-use efficiency under integrated use of nano urea, this study was undertaken during the rabi seasons of 2021–22 and 2022–23 at the Uttar Banga Krishi Viswavidyalaya, Pundibari, West Bengal. The experiment was laid-out in a randomized block design (RBD) having 13 treatments consisting of foliar application of water, nano urea (once at tillering and twice at tillering and jointing) and 5% urea in combination with each of 50,75 and 100% of recommended nitrogen doses (RDN) along with a control (without N), and each treatment was replicated thrice. It was observed that foliar application of nano urea (0.4%) twice at tillering and jointing stages with 100% (RDN 5,759 kg/ha) was statistically at par with 100% RDN + 5% foliar spray of urea twice at tillering and jointing stage (5,985 kg/ha) in terms of grain yield, the latter being the highest yielder. Foliar application of either urea or nano urea by curtailing 25-50% RDN did not brought about superior growth and yield performances. Under each level of RDN (100%, 75% and 50%), foliar application of 5% urea was a bit superior to nano urea, though the difference was not statistically significant. However, nano urea reflected higher nitrogen-use efficiency under all levels RDN; partial factor productivity of nitrogen (PFPN) and agronomic nitrogen-use efficiency (ANUE) values be- ing the highest under 50% RDN along with two spraying of nano urea (56.76 and 24.50 kg grain/kg N). The treat- ment comprising 100% RDN+ 5% spray of urea at tillering and jointing stages resulted in the highest net return (69608 `/ha) and B: C ratio (2.58), closely followed by the treatment comprising nano urea application twice (gross return, net return and B: C ratio of 109,426 `/ha, 63,748 `/ha and 2.40, respectively) with 100% RDN, being statis- tically at par with each other. It can be concluded from this two-year trial that only foliar application of nano urea by curtailing soil nitrogen application to 25-50% resulted in reduced yield performances of wheat; nano urea had pro- motive effects on yields when it was applied along with 100% RDN.

B-184 Purifying High-Quality Genomic DNA From Frozen Blood Samples Stored up to 1.5 Years at -20°C

Abstract Background Maintaining DNA integrity in blood samples, from transport to storage to processing, is crucial to the success of downstream applications, especially regarding diagnostic applications. Additionally, due to the invasive nature of collecting blood samples, minimizing resampling to avoid excess patient pain and cost of collection and transportation are important to consider. Having the option to store and maintain portions of blood samples in the freezer enables future retesting without recollection. Since -20°C freezers generally are more affordable, require less energy, and have smaller footprints than -80°C freezers, demonstrating frozen blood stability at -20°C temperatures could decrease sampling burdens for labs with facility constraints. Methods Whole blood was collected into 9mL vacuum tubes (HemaSure-OMXP) from healthy donors. Aliquots of whole blood and buffy coat samples were stored at -20°C and thawed for genomic DNA extractions (QIAamp DNA Blood Mini Kit, Qiagen) at a range of different timepoints; frozen whole blood samples were processed at 0, 49, 141, and 148 days and frozen buffy coat samples were processed at 7, 27, 36, 185, 246, 373, 394, 584, and 604 days. Purified gDNA was quantified with Qubit (Broad Range dsDNA Assay, ThermoFisher) and qualified with Nanodrop One Microvolume UV-Vis Spectrophotometer (ThermoFisher) and TapeStation (Genomic DNA ScreenTape, Agilent Technologies). Results No significant decrease was observed for DNA yield and quality between fresh and frozen whole blood samples. Average DNA yields from 200 μL whole blood were 4.5 ± 2.2 μg and 5.3 ± 1.2 μg for fresh and frozen blood respectively. Average DNA integrity (DIN) was 6.9 ± 0.4 and 7.1 ± 0.3 for fresh and frozen blood respectively. Donor differences in blood samples can help explain the variance in DNA yields reported. No significant decrease was observed for DNA yield and quality in buffy coat samples frozen for longer at -20°C. In total, 84 whole blood and 25 buffy coat samples were processed in this study. Conclusions Although buffy coat samples tended to have higher yields than whole blood, due to the higher amount of white blood cells present, both sample types provided sufficient amounts of gDNA for downstream applications. Freezing blood samples for future testing can be effective at reducing the need for resampling without sacrificing the nucleic acid yield or quality of fresh samples. Additionally, as technological advancements are made for equipment and assays, frozen blood samples can be retested and compared to previous results for improved diagnostic decisions, done without requiring recollection from labs and patients.