The Increased aspartate levels in transgenic cotton (Gossypium hirsutum L.) lead to improved tolerance against whitefly (Bemisia tabaci, Gennadius).

The whitefly, a polyphagous insect pest feeding on nearly 1328 plant species, is a major threat to global cotton production and incurs up to 50% yield losses in cotton production in Pakistan. We investigated whether increased aspartate in phloem sap imparts whitefly toxicity and protects cotton plants from intense damage. The enzymatic step for aspartate production is carried through aspartate aminotransferase (AAT). In this study, we constitutively overexpressed the Oryza sativa cytoplasmic AAT (OsAAT2) under the CaMV35S promoter in Gossypium hirsutum cv. CIM-482. Real-time PCR analysis of the AAT transcripts revealed a 2.85- to 31.7-fold increase in mRNA levels between the different cotton lines. A substantial increase in the free-amino acid content of the major N-assimilation and transport amino acids (aspartate, glutamate, asparagine, and glutamine) was seen in the phloem sap of the transgenic cotton lines. The bioassay revealed that the two transgenic cotton lines with the highest free aspartate content in the phloem sap exhibited 97 and 94% mortality in the adult whitefly population and a 98 and 96% decline in subsequent nymph populations, respectively. There was also a significant change in the physiological behaviour of the transgenic cotton lines, with an increased net assimilation (A), gaseous exchange (Gs) and rate of transpiration (E). Improved morphological characteristics like plant height, total number of bolls and fiber yield were recorded in transgenic cotton lines. The AAT gene shows promise in mitigating whitefly infestations and enhancing the overall health and yield of cotton plants.

Discovery of new molecular hybrid derivatives with coumarin scaffold bearing pyrazole/oxadiazole moieties: Molecular docking, POM analyses, in silico pharmacokinetics and in vitro antimicrobial evaluation with identification of potent antitumor pharmacophore sites.

This synthetic organic methodology involves the creation of novel coumarin-based hybrids of series (1-4) with pyrazole ring and (5-8) with oxadiazole moiety. The targeted compounds were tested for In vitro Antimicrobial efficacy against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans pathogenic microbes using disc diffusion and broth microdilution with ciprofloxacin and fluconazole as reference standards. Density functional theory (DFT) studies were used to study atomic structure and reactivity, including absolute electronegativity (χ), electrophilicity (ω), electron acceptor (ω+), donor capabilities (ω-), electron affinity (EA), energy gap (ΔE), global hardness (η), global softness (S), and ionisation potential (IP) and FMOs, NBOs, MEP, and Mulliken Charge analysis. The POM tests found three integrated pharmacophore sites with antibacterial, antiviral, and anticancer activities. Molecular docking studies are also used to determine the S. aureus nucleoside diphosphate kinase receptor's affinity and mode of action for the synthesized drugs. In silico analysis of thermodynamic and therapeutic effectiveness properties, including Lipinski's 'rule of five', Veber's rule, and ADME properties, predicted toxicity-free, non-carcinogenic, and risk-free oral administration of the synthesized complexes.

Developing a simple and efficient modeling solution for predicting key phenological stages of table grapes in a non-traditional viticulture zone in south Asia.

Phenological shifts are one of the most visible signs of climatic variability and change in the biosphere. However, modeling plant phenological responses has always been a key challenge due to climatic variability and plant adaptation. Grapevine is a phenologically sensitive crop and, thus, its developmental stages are affected by the increase in temperature. The goal of this study was to develop a temperature-based grapevine phenology model (GPM) for predicting key developmental stages for different table grape cultivars for a non-traditional viticulture zone in south Asia. Experiments were conducted in two vineyards at two locations (Chakwal and Islamabad) in the Pothawar region of Pakistan during the 2019 and 2020 growing seasons for four cultivars including Perlette, King's Ruby, Sugraone and NARC Black. Detailed phenological observations were obtained starting in January until harvest of the grapes. The Mitscherlich monomolecular equation was used to develop the phenology model for table grapes. There was a strong non-linear correlation between the Eichhorn and Lorenz phenological (ELP) scale and growing degree days (GDD) for all cultivars with coefficient of determinations (R2) ranging from 0.90 to 0.94. The results for model development indicated that GPM was able to predict phenological stages with high skill scores, i.e., a root mean square (RMSE) of 2.14 to 2.78 and mean absolute error (MAE) of 1.86 to 2.26 days. The prediction variability of the model for the onset timings of phenological stages was up to 3 days. The results also reveal that the phenology model based on GDD approach provides an efficient planning tool for viticulture industry in different grape growing regions. The proposed methodology, being a simpler one, can be easily applied to other regions and cultivars as a predictor for grapevine phenology.

CSM-CROPGRO model to simulate safflower phenological development and yield.

Crop simulation models are valuable tools for decision making regarding evaluation and crop improvement under different field conditions. CSM-CROPGRO model integrates genotype, environment and crop management portfolios to simulate growth, development and yield. Modeling the safflower response to varied climate regimes are needed to strengthen its productivity dynamics. The main objective of the study was to evaluate the performance of DSSAT-CSM-CROPGRO-Safflower (Version 4.8.2) under diverse climatic conditions. The model was calibrated using the field observations for phenology, biomass and safflower grain yield (SGY) of the year 2016-17. Estimation of genetic coefficients was performed using GLUE (Genetic Likelihood Uncertainty Estimation) program. Simulated results for days to flowering, maturity, biomass at flowering and maturity and SGY were predicted reasonably with good statistical indices. Model evaluation results elucidate phenological events with low root mean square error (6.32 and 6.52) and high d-index (0.95 and 0.96) for days to flowering and maturity respectively for all genotypes and climate conditions. Fair prediction of safflower biomass at flowering and maturity showed low RMSE (887.3 and 564.3 kg ha-1) and high d-index (0.67 and 0.93) for the studied genotypes across the environments. RMSE for validated safflower grain yield (101.8 kg ha-1) and d-index (0.95) depicted that model outperformed for all genotypes and growing conditions. Longer appropriate growing conditions at NARC-Islamabad took optimal duration to assimilate photosynthetic products lead to higher grain yield. Safflower resilience to different environments showed that it can be used as an alternate crop for different agroecological regions. Furthermore, CROPGRO-Safflower model can be used as tool to further evaluate inclusion of safflower in the existing cropping systems of studied regions.

Kappa-carrageenan and xylooligosaccharide effect on water mobility and structural changes in silver carp proteins during frozen storage.

BACKGROUND: The cryoprotective effect of xylooligosaccharide (XO) and kappa-carrageenan (KC) mixture on silver carp proteins in fluctuated frozen storage from 4 to -18 °C was analyzed. Positive control as a conventional cryoprotectant mixture of sucrose (4%) and sorbitol (4%), KC (3%) and XO/KC (3%) treatments were incorporated in silver carp surimi and myofibrillar proteins to analyze the water mobility and its influence on structural attributes. RESULTS: The temperature fluctuation significantly increased the structural alteration in samples with no treatments due to oxidative changes, protein denaturation and recrystallization. Meanwhile, the mixture of XO and KC (XO/KC 3%) significantly reduced the tertiary and secondary structural alterations by preventing the oxidative changes in α-helix and tryptophan (Trp) residues. Moreover, XO/KC (3%) inhibited water mobility, hindering the T22 relaxation time, as compared to the samples added with KC (3%) and the positive control. Interestingly, the XO/KC (3%) mixture significantly reduced the formation of extracellular spaces and recrystallization by restricting the partial dehydration of muscles and extracellular solution concentration. CONCLUSION: From the current results, it can be concluded that the XO/KC mixture could be efficient in protecting aquatic food proteins during fluctuating frozen storage by preventing the exposure of Trp residues and α-helix contents. Moreover, XO/KC restricted the water mobility by establishing a bond and making water unavailable for crystallization and recrystallization. Therefore, XO/KC could be used as an effective mixture to prevent fluctuated and frozen storage changes in aquatic foods. © 2024 Society of Chemical Industry.

Exploring the natural efficacy of spirulina powder for combating obesity, diabetes, and inflammation.

BACKGROUND: An increasing incidence of metabolic disorders emphasizes the need to explore natural treatments. Spirulina, a microalga with a rich nutrient profile, offers a promising solution for obesity, diabetes, and inflammation. This study provides a meticulous analysis of spirulina powder, evaluating its physicochemical attributes and technofunctional properties through the use of advanced analytical techniques. RESULTS: Spirulina powder demonstrated strong flowability, substantial water and oil absorption capacity, and moderate foaming characteristics. The ethanolic extract of spirulina was found to be a repository of phenolic (6.93 mg GAE/g) and flavonoid (7.17 mg QE/g) compounds, manifesting considerable antioxidant activity with a 58.49 g kg-1 inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals. The extract also exhibited pronounced inhibitory effects on lipase and amylase enzymes, with inhibition percentages of 72.05 g kg-1 and 70.28 g kg-1, respectively, and displayed a glucose retention capacity of 1.28 mg dL-1 (68.52 g kg-1) in a dialysis membrane assay. These results suggest its efficacy in modulating obesity and glycemic control. The powder also showed a potent anti-inflammatory response by mitigating protein denaturation. CONCLUSION: Spirulina powder is a potent natural agent with multiple health benefits, meriting its incorporation into functional foods. It could be suitable for application in the food industry, offering a natural strategy to combat metabolic diseases. This research adds to the scientific literature on spirulina, paving the way for future research into its utilization. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Clinical Outcomes in a Randomized Controlled Trial Comparing Point-of-Care With Standard Human Immunodeficiency Virus (HIV) Viral Load Monitoring in Nigeria.

BACKGROUND: Point-of-care (POC) viral load (VL) tests provide results within hours, enabling same-day treatment interventions. We assessed treatment outcomes with POC vs standard-of-care (SOC) VL monitoring. METHODS: We implemented a randomized controlled trial at an urban and rural hospital in Nigeria. Participants initiating antiretroviral therapy (ART) were randomized 1:1 for monitoring via the POC Cepheid Xpert or SOC Roche COBAS (v2.0) HIV-1 VL assays. Viral suppression (VS) and retention in care at 12 months were compared via intention-to-treat (ITT) and per-protocol (PP) analyses. Post-trial surveys for POC patients and healthcare workers (HCWs) evaluated acceptability. RESULTS: During April 2018-October 2019, 268 SOC and 273 POC patients enrolled in the trial. Viral suppression at <1000 copies/mL at 12 months was 59.3% (162/273) for POC and 52.2% (140/268) for SOC (P = .096) in ITT analysis and 77.1% (158/205) for POC and 65.9% (137/208) for SOC (P = .012) in PP analysis. Retention was not significantly different in ITT analysis but was 85.9% for POC and 76.9% for SOC (P = .02) in PP analysis. The increased VS in the POC arm was attributable to improved retention and documentation of VL results. POC monitoring was preferred over SOC by 90.2% (147/163) of patients and 100% (15/15) of HCWs thought it facilitated patient care. CONCLUSIONS: POC VL monitoring did not improve 12-month VS among those with results but did improve retention and VS documentation and was preferred by most patients and HCWs. Further research can inform best POC implementation conditions and approaches to optimize patient care. CLINICAL TRIALS REGISTRATION: NCT03533868.

Timeliness of Point-of-Care Viral Load Results Improves Human Immunodeficiency Virus Monitoring in Nigeria.

BACKGROUND: Human immunodeficiency virus (HIV) viral load (VL) monitoring is critical for antiretroviral therapy (ART) management. Point-of-care (POC) VL testing has been reported to be feasible and preferred over standard-of-care (SOC) testing in many low- and middle-income country settings where rapid results could improve patient outcomes. METHODS: The timeliness of receipt of VL results was evaluated in an open-label, randomized, controlled trial among patients newly initiating ART. Clinical outcomes with POC VL monitoring using Cepheid Xpert vs SOC VL at Jos University Teaching Hospital and Comprehensive Health Centre Zamko in Nigeria were assessed. We determined time between specimen collection and recording of VL in patient charts, receipt of results, and ART switch for those who met virologic failure criteria. RESULTS: Between April 2018 and October 2019, we screened 696 ART-naive individuals; 273 were randomized to POC and 268 to SOC HIV-1 VL testing. Participants in the POC arm received VL results significantly faster than those in the SOC arm (0.1 median days, interquartile range [IQR], 0.1-0.2 vs 143.1 days, IQR, 56.0-177.1, respectively; P < .0001). Participants in the POC arm with confirmed virologic failure vs those in the SOC arm were switched more rapidly to a second-line regimen (0 median days, IQR, 0-28 vs 66 days, IQR, 63-123, respectively; P = .03). CONCLUSIONS: POC VL testing resulted in significant improvement in the timeliness of VL result receipt by patients and use for effective HIV clinical management. In patients experiencing VL failure, POC monitoring enabled prompt switching to second-line ART regimens. CLINICAL TRIALS REGISTRATION: NCT03533868.

Sulfonylpyrazole- and pyrazole-directed ortho-selective C-H functionalization/alkenylation and desulfenylative olefination of aryl(sulfonyl)pyrazoles.

ortho-Selective C-H alkenylation of arenes was achieved using sulfonylpyrazoles and pyrazoles as directing groups, favored by a combination of a Pd(OAc)2 catalyst, Boc-Sar-OH and silver acetate. A wide variety of mono-alkenylated products of aryl-sulfonylpyrazoles and pyrazoles were synthesized with complete site-selectivity under mild reaction conditions. This transformation tolerated several electron-withdrawing and electron-donating groups on the aryl ring and the yields ranged from 52% to 70%, producing highly decorated/valuable alkenylated sulfonylpyrazole and pyrazole derivatives. Amazingly, switching of the oxidant, with the use of AgBF4 in place of AgOAc, offered cinnamic acid derivatives through de-sulfonylation followed by alkenylation at the same position with good yields in the case of aryl-sulfonylpyrazoles. These kinds of molecules have great biological importance and target predictions indicate that they may serve as potential antifungal and anti-tumor agents.

Photosystems and antioxidative system of rye, wheat and triticale under Pb stress.

Lead (Pb2+) pollution in the soil sub-ecosystem has been a continuously growing problem due to economic development and ever-increasing anthropogenic activities across the world. In this study, the photosynthetic performance and antioxidant capacity of Triticeae cereals (rye, wheat and triticale) were compared to assess the activities of antioxidants, the degree of oxidative damage, photochemical efficiency and the levels of photosynthetic proteins under Pb stress (0.5 mM, 1 mM and 2 mM Pb (NO3)2). Compared with triticale, Pb treatments imposed severe oxidative damage in rye and wheat. In addition, the highest activity of major antioxidant enzymes (SOD, POD, CAT, and GPX) was also found to be elevated. Triticale accumulated the highest Pb contents in roots. The concentration of mineral ions (Mg, Ca, and K) was also high in its leaves, compared with rye and wheat. Consistently, triticale showed higher photosynthetic activity under Pb stress. Immunoblotting of proteins revealed that rye and wheat have significantly lower levels of D1 (photosystem II subunit A, PsbA) and D2 (photosystem II subunit D, PsbD) proteins, while no obvious decrease was noticed in triticale. The amount of light-harvesting complex II b6 (Lhcb6; CP24) and light-harvesting complex II b5 (Lhcb5; CP26) was significantly increased in rye and wheat. However, the increase in PsbS (photosystem II subunit S) protein only occurred in wheat and triticale exposed to Pb treatment. Taken together, these findings demonstrate that triticale shows higher antioxidant capacity and photosynthetic efficiency than wheat and rye under Pb stress, suggesting that triticale has high tolerance to Pb and could be used as a heavy metal-tolerant plant.

Temperature variability during the growing season affects the quality attributes of table grapes in Pothwar-insight from a new emerging viticulture region in South Asia.

Rising air temperature due to climate change has posed a mammoth challenge to global viticulture and key berry quality traits are compromised. Exploring the effects of seasonal temperature variability on berry ripening and quality attributes in different viticulture regions may help in sustainable viticulture industry. The present research was designed to explore the effect of temperature variables on key quality attributes of table grape cultivars in Pothwar region of Pakistan. Key berry quality traits such as total soluble solids (TSS), titratable acidity (TA), maturity indices (MI), ascorbic acid, sugars, total polyphenol contents (TPC) and total anthocyanin contents (TAC) were unlocked for four important table grape cultivars under varying environmental conditions at Chakwal and Islamabad districts for two consecutive vintages of 2019 and 2020. The district Chakwal has up to 0.92 °C, 1.35 °C, 1.12°C and 0.81°C higher Tmin, Tmax, Tmean and diurnal temperature variation (DTV) respectively, compared to Islamabad particularly for the 2019 vintage. The results of the present study revealed that the warmer site (Chakwal) has significantly (P ≤0.05) higher juice pH, TSS (°brix) and maturity indices (MI) particularly for the relatively hotter vintage of 2019. Interestingly, MI was 33% higher for the relatively warmer vintage of 2019 compared to 2020 with relatively lower acidity (up to 38%). Moreover, higher titratable acidity (11.2%), ascorbic acid (28.5%), polyphenols (20.3%) and anthocyanins (10.6%) were noticed for the colder Islamabad compared to Chakwal. Although elevated temperature for warmer location and vintage favoured berry ripening, however key biochemical attributes such as titratable acidity, ascorbic acid, polyphenols and anthocyanins were negatively affected. The findings of the present research provide useful insight into the impact of growing season temperature on key berry attributes and may help devise adaptation strategies to improve berry quality.

Seasonal variability in the effect of temperature on key phenological stages of four table grapes cultivars.

Progressive warming of the grape growing regions has reduced the land capability for sustainable grapevine production and the geographical distribution of grapes. Bud burst, blooming, berry set, veraison, and harvest are the key phenological stages of grapevine, and are crucial for managing vineyard activities. The objective of this study was to evaluate the effect of seasonal temperature variability on the timing of key phenological stages of table grape cultivars in a new emerging viticulture region, i.e., the Pothwar region of Pakistan. Phenological stages of four table grape cultivars were recorded during two consecutive growing seasons at two locations. All phenological stages were attained earlier for the relatively warmer location, i.e., Chakwal. Similarly, the length of the growing season from bud burst to harvest was 15 to 21 days longer for the 2020 growing season than for the 2019 growing season, which corresponds to the inter-annual temperature variability. Moreover, the grapevine cultivars showed a distinct response for each growth phase; cv. Perlette matured earlier while cv. NARC Black was the last to ripen. Despite the large variability in the length of the active growing period from bud burst to harvest, accumulated growing degree days (GDD) varied only in a narrow range, i.e., 1510-1557 for cv. Perlette, 1641-1683 for cv. King's Ruby, 1744-1770 for cv. Sugraone, and 1869-1906 for cv. NARC Black. This implies that seasonal temperature variability using GDD is a better indicator for the phenology of table grape cultivars compared to regular time. It is clear from the results from this study that the variation in phenological responses of table grape cultivars due to temperature differences necessitates genotype and site-specific vineyard management.

Simulation of winter wheat response to variable sowing dates and densities in a high-yielding environment.

Crop multi-model ensembles (MME) have proven to be effective in increasing the accuracy of simulations in modelling experiments. However, the ability of MME to capture crop responses to changes in sowing dates and densities has not yet been investigated. These management interventions are some of the main levers for adapting cropping systems to climate change. Here, we explore the performance of a MME of 29 wheat crop models to predict the effect of changing sowing dates and rates on yield and yield components, on two sites located in a high-yielding environment in New Zealand. The experiment was conducted for 6 years and provided 50 combinations of sowing date, sowing density and growing season. We show that the MME simulates seasonal growth of wheat well under standard sowing conditions, but fails under early sowing and high sowing rates. The comparison between observed and simulated in-season fraction of intercepted photosynthetically active radiation (FIPAR) for early sown wheat shows that the MME does not capture the decrease of crop above ground biomass during winter months due to senescence. Models need to better account for tiller competition for light, nutrients, and water during vegetative growth, and early tiller senescence and tiller mortality, which are exacerbated by early sowing, high sowing densities, and warmer winter temperatures.

Genetic characterization of flowering and phytochrome genes in peanut (Arachis hypogaea L.) for early maturity.

BACKGROUND: Peanut (Arachis hypogaea L.) production and cropping pattern is highly influenced by the climatic factors including temperature and rain pattern fluctuations. It is one of the most important cash crop in the rain fed areas of Pakistan and its production, under changing climatic conditions, that can be improved by developing short duration varieties. The present study was based on the molecular characterization of the maturity associated gene families in the peanut under two light conditions. METHODS AND RESULTS: Genomic analysis based on the in silico study of important gene families for early maturity associated attributes like flowering time, their pattern, duration and photoperiodism was done for a comprehensive mapping of maturity related genes. Phytochromes genes Phy A, Phy B and Phy E and flowering genes FT2a, Ft5a and COL2 were selected for in silico characterization for protein based analysis including Multiple Sequence Alignment (MSA), and Neighbor Joining (NJ) tree. MSA and NJ trees of the peanut with Arabidopsis thaliana and Glycine max showed a clear picture of the phylogenetic relationship on the basis of selected gene proteins. Expression profile of phytochrome and flowering genes revealed that photoperiod conditions i.e. short and long days, have great influence on the Phy A, Phy B and Phy E, Ft2a, FT5a and COL2 gene expression pattern. In current study, the relative expression of all studied genes was found higher in short day light condition at flower initiation stage of the plants than in the long light day condition with exception of COL2 gene protein. CONCLUSIONS: The molecular characterization based on the in silico study of the particular genes and qPCR based gene expression profiling of the selected genes provided an evidence of the role of these genes and their comparative analysis under two photoperiodic conditions.

Optimizing sowing window, cultivar choice, and plant density to boost maize yield under RCP8.5 climate scenario of CMIP5.

The impacts of climate change and possible adaptations to food security are a global concern and need greater focus in arid and semi-arid regions. It includes scenario of Coupled Model Intercomparison Phase 5 (CMIP-RCP8.5). For this purpose, two DSSAT maize models (CSM-CERES and CSM-IXIM) were calibrated and tested with two different maize cultivars namely Single Cross 10 (SC10) and Three Way Cross 324 (TW24) using a dataset of three growing seasons in Nile Delta. SC10 is a long-growing cultivar that is resistant to abiotic stresses, whereas TW24 is short and sensitive to such harsh conditions. The calibrated models were then employed to predict maize yield in baseline (1981-2010) and under future time slices (2030s, 2050s, and 2080s) using three Global Climate Models (GCMs) under CMIP5-RCP8.5 scenario. In addition, the use of various adaptation options as shifting planting date, increasing sowing density, and genotypes was included in crop models. Simulation analysis showed that, averaged over three GCMs and two crop models, the yield of late maturity cultivar (SC10) decreased by 4.1, 17.2, and 55.9% for the three time slices of 2030s, 2050s, and 2080s, respectively, compared to baseline yield (1981-2010). Such reduction increased with early maturity cultivar (TW24), recording 12.4, 40.6, and 71.3% for near (2030s), mid (2050s), and late century (2080s) respectively relative to baseline yield. The most suitable adaptation options included choosing a stress-resistant genotype, changing the planting date to plus or minus 30 days from baseline planting date, and raising the sowing density to 9 m-2 plants. These insights could minimize the potential reduction of climate change-induced yields by 39% by late century.

In vitro evaluation of novel (nanoparticle) oral delivery systems allow selection of gut immunomodulatory formulations.

Oral delivery is the most convenient way to vaccinate cultured fish, however it is still problematic, primarily due to a lack of a commercially valid vaccine vehicle to protect the antigen against gastric degradation and ensure its uptake from the intestine. With the goal of advancing the potential to vaccinate orally, this study evaluates a novel silicon nanoparticle-based vehicle (VacSaf carrier). Aeromonas salmonicida antigens were formulated with the VacSaf carrier using different preparation methods to generate dry powder and liquid formulations. Twelve formulations were first subjected to an in vitro evaluation where the A. salmonicida bacterin conjugated to VacSaf carriers were found superior at inducing pro-inflammatory cytokine expression in primary leucocyte cultures and the macrophage/monocyte cell line RTS-11 compared with A. salmonicida bacterin alone. This was especially apparent after exposure to acid conditions to mimic stomach processing. One formulation (FD1) was taken forward to oral delivery using two doses and two administration schedules (5 days vs 10 days, the latter 5 days on, 5 days off, 5 days on), and the transcript changes of immune genes in the intestine (pyloric caeca, midgut and hindgut) and spleen were evaluated by qPCR and serum IgM was measured by ELISA. The VacSaf carrier alone was shown to be safe for use in vivo, in that no side-effects were seen, but it did induce expression of some cytokines, and may have value as an oral adjuvant candidate. The FD1 bacterin formulation was effective at inducing a range of cytokines associated with innate and adaptive immunity, mainly in the pyloric caeca, compared to A. salmonicida bacterin alone (which had almost no effect), and confirms the immune competence of this gut region following appropriate oral vaccination. These results reveal that in vitro screening of formulations for oral delivery has value and can be used to assess the most promising formulations to test further.

Development of broad-spectrum and sustainable resistance in cotton against major insects through the combination of Bt and plant lectin genes.

KEY MESSAGE: Second generation Bt insecticidal toxin in comibination with Allium sativum leaf agglutinin gene has been successfully expressed in cotton to develop sustainable resistance against major chewing and sucking insects. The first evidence of using the Second-generation Bt gene in combination with Allium sativum plant lectin to develop sustainable resistance against chewing and sucking insects has been successfully addressed in the current study. Excessive use of Bt δ-endotoxins in the field is delimiting its insecticidal potential. Second-generation Bt Vip3Aa could be the possible alternative because it does not share midgut receptor sites with any known cry proteins. Insecticidal potential of plant lectins against whitefly remains to be evaluated. In this study, codon-optimized synthetic Bt Vip3Aa gene under CaMV35S promoter and Allium sativum leaf agglutinin gene under phloem-specific promoter were transformed in a local cotton variety. Initial screening of putative transgenic cotton plants was done through amplification, histochemical staining and immunostrip assay. The mRNA expression of Vip3Aa gene was increased to be ninefold in transgenic cotton line L6P3 than non-transgenic control while ASAL expression was found to be fivefold higher in transgenic line L34P2 as compared to non-transgenic control. The maximum Vip3Aa concentration was observed in transgenic line L6P3. Two copy numbers in homozygous form at chromosome number 9 and one copy number in hemizygous form at chromosome number 10 was observed in transgenic line L6P3 through fluorescent in situ hybridization. Significant variation was observed in transgenic cotton lines for morphological characteristics, whereas physiological parameters of plants and fiber characteristics (as assessed by scanning electron microscopic) remained comparable in transgenic and non-transgenic cotton lines. Leaf-detach bioassay showed that all the transgenic lines were significantly resistant to Helicoverpa armigera showing mortality rates between 78% and 100%. Similarly, up to 95% mortality of whiteflies was observed in transgenic cotton lines when compared with non-transgenic control lines.

Recycling of sugar crop disposal to boost the adaptation of canola (Brassica napus L.) to abiotic stress through different climate zones.

We need to produce higher foods even under declining natural resources to feed the projected population of 9 billion by 2050 and to sustain food security and nutrition. Abiotic stress has adversely affected canola crop and oil quality especially in sandy soils. To combat this stress, adaptation at the farm level using new and cost-effective amendments are required. Field trials were conducted in two different climatic zones to determine the efficacy of cane molasses, bagasse ash, sugar beet factory lime, and their compost mixtures to improve soil quality and heat stress-adapting canola. The results showed a significant improvement in bulk density, hydraulic conductivity, organic matter content, and available macronutrients of sandy soil and subsequent canola growth, yield, quality and water productivity due to the application of the tested soil amendments, particularly those mixed with compost. Despite the estimated reduction of yield by 18.5% due to heat stress, application of sugar beet lime and compost mixture not only compensated for this reduction but also increased the seed yield by 27.0%. These findings highlight the value of recycling compost-based sugar crop disposal as a cost-effective technology to boost crop tolerance to abiotic stress, ensuring sustainable agriculture and food security in arid environments.

Low levels of HIV-1 drug resistance mutations in patients who achieved viral re-suppression without regimen switch: a retrospective study.

BACKGROUND: We identified a HIV-positive cohort in virologic failure (VF) who re-suppressed without drug switch. We characterized their drug resistance mutations (DRM) and adherence profiles to learn how to better manage HIV drug resistance. A retrospective cohort study utilizing clinical data and stored samples. Patients received ART at three Nigerian treatment centres. Plasma samples stored when they were in VF were genotyped. RESULT: Of 126 patients with samples available, 57 were successfully genotyped. From ART initiation, the proportion of patients with adherence ≥90% increased steadily from 54% at first high viral load (VL) to 67% at confirmed VF, and 81% at time of re-suppressed VL. Sixteen (28%) patients had at least one DRM. Forty-six (81%) patients had full susceptibility to the three drugs in their first-line (1 L) regimen. Thirteen (23%) were resistant to at least one antiretroviral drug but three were resistant to drugs not used in Nigeria. Ten patients had resistance to their 1 L drug(s) and six were fully susceptible to the three drugs in the recommended second-line regimen. CONCLUSION: This cohort had little drug resistance mutations. We conclude that if adherence is not assured, patients could exhibit virologic failure without having developed mutations associated with drug resistance.

Identification and validation of superior housekeeping gene(s) for qRT-PCR data normalization in Agave sisalana (a CAM-plant) under abiotic stresses.

The adaptive mechanisms in Agave species enable them to survive and exhibit remarkable tolerance to abiotic stresses. Quantitative real-time PCR is a highly reliable approach for validation of targeted differential gene expression. However, stable housekeeping gene(s) is prerequisite for accurate normalization of expression data by qRT-PCR. Till date, no systematic validation study for candidate housekeeping gene identification or evaluation has been carried-out in Agave species. A total of 17 candidate housekeeping genes were identified from the de novo assembled transcriptomic data of A. sisalana and rigorously analyzed for expression stability assessment under drought, heat, cold and NaCl stress. Different statistical algorithms like geNorm, BestKeeper, NormFinder, and RefFinder on expression data determined the superior housekeeping gene(s) for accurate normalization of the gene of interest (GOI). The comprehensive evaluation revealed the ß-Tub 4, WIN-1 and CYC-A as the most stable, while EEF1α, GAPDH, and UBE2 were ranked as the least stable genes in pooled samples. Pairwise combination by geNorm showed that up to two housekeeping genes would be adequate to normalize the GOI expression data precisely. Validation of identified most and least stable housekeeping genes was carried-out by normalizing the expression data of AsHSP20 under abiotic stress conditions. Copy number of AsHSP20 gene supports the reliability of the genes used for normalization. This is the first report on the screening and validation of the housekeeping genes under abiotic stress condition in A. sisalana that would assist to understand the stress tolerance mechanisms by novel gene identification and accurate validation.