Fresh Leaf Research Articles

Zinc Oxide Nanoparticles Alleviate Salt Stress in Cotton (Gossypium hirsutum L.) by Adjusting Na+/K+ Ratio and Antioxidative Ability.

Soil salinization poses a threat to the sustainability of agricultural production and has become a global issue. Cotton is an important cash crop and plays an important role in economic development. Salt stress has been harming the yield and quality of many crops, including cotton, for many years. In recent years, soil salinization has been increasing. It is crucial to study the mechanism of cotton salt tolerance and explore diversified materials and methods to alleviate the salt stress of cotton for the development of the cotton industry. Nanoparticles (NPs) are an effective means to alleviate salt stress. In this study, zinc oxide NPs (ZnO NPs) were sprayed on cotton leaves with the aim of investigating the intrinsic mechanism of NPs to alleviate salt stress in cotton. The results show that the foliar spraying of ZnO NPs significantly alleviated the negative effects of salt stress on hydroponic cotton seedlings, including the improvement of above-ground and root dry and fresh weight, leaf area, seedling height, and stem diameter. In addition, ZnO NPs can significantly improve the salt-induced oxidative stress by reducing the levels of MDA, H2O2, and O2- and increasing the activities of major antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Furthermore, RNA-seq showed that the foliar spraying of ZnO NPs could induce the expressions of CNGC, NHX2, AHA3, HAK17, and other genes, and reduce the expression of SKOR, combined with the CBL-CIPK pathway, which alleviated the toxic effect of excessive Na+ and reduced the loss of excessive K+ so that the Na+/K+ ratio was stabilized. In summary, our results indicate that the foliar application of ZnO NPs can alleviate high salt stress in cotton by adjusting the Na+/K+ ratio and regulating antioxidative ability. This provides a new strategy for alleviating the salt stress of cotton and other crops, which is conducive to the development of agriculture.

Effects of Leaf Species and Conditioning State of Fresh Leaves on Colonization by Stream and Pond Macroinvertebrates

Fresh, green leaves are increasing as resources in aquatic ecosystems due to more frequent severe spring and summer storms, but research on allochthonous resources typically uses senescent leaves. We examined macroinvertebrate colonization of green leaves of three native deciduous trees (red maple, red oak, and tulip poplar) over two weeks within both a stream and fishless pond. Leaf colonization varied depending on the taxa of leaves and colonizers, submersion time, and the ecosystem examined. Within the stream, the densities of isopods (Lirceus sp.) and snails (mostly the invasive Potamopyrgus antipodarum) did not vary significantly across leaf species. In contrast, mayflies (Tricorythodes sp.) in the stream colonized tulip poplar in greater numbers than red oak leaves, while higher densities of planarians (order Tricladida) occurred within red oak leaves. The numbers of mayflies and snails decreased significantly by the second week, but the densities of isopods and planaria within stream leaf packs were consistent. In contrast, within the pond, significantly more isopods (Caecidotea communis) were collected after the first than after the second week of submersion and in tulip poplar leaves. Clams (Sphaeriidae) in the pond, on the other hand, were more prevalent in the second week but did not discriminate among leaf species. While the number of leeches (mostly Helobdella stagnalis) was consistent across weeks, significantly fewer leeches resided within tulip poplar leaves than within red oak leaves. Our results suggest that there are no consistent colonization-preference rankings of species of fresh leaves across freshwater benthic macroinvertebrate taxa. Even within a functional feeding group (e.g., the two types of isopods and snails, all detritivore-herbivores), there were differences in colonization patterns. Therefore, increased allochthonous inputs of fresh leaf litter due to severe spring- and summer-time storms are likely to promote the populations of various taxa to different extents.

The potency of fungal entomopathogens isolated from Spodoptera frugiperda as endophytic plant-growth promoter

Previous studies have identified endophytic fungi as plant growth promoters and insect pest management. In the current study, six fungal isolates of Metarhizium anisopliae and Beauveria bassiana were isolated from larval corpses of fall armyworm (Spodoptera frugiperda) and tested for their ability to colonize leaf tissue through root and leaf applications or foliar-spray and to promote the growth of young maize (Zea mays) plants. The maize growth attributes examined were germination rate, height, dry and fresh weight, and leaf area. The research was conducted in May until June 2023 in Universitas Sriwijaya, Indonesia. The results revealed that M. anisopliae and B. bassiana were isolated from the cadavers of S. frugiperda and confirmed to be endophytic fungi. Root application was more effective to enhance the fresh weight, plant height, and leaf area of maize compared to foliar-spray. The fungi could significantly enhance the fresh weight, plant height, and leaf area of the young maize plants. The best isolate that could enhance the maize growth was the B. bassiana WTTJC260521A isolate. These findings suggest that M. anisopliae and B. bassiana isolated from fall armyworm corpses that had the potential to enhance maize plant growth through root applicaton.

التخليق الحيوي لجسيمات الفضة النانوية باستخدام مستخلصات الشاي الأخضر

اصبح تركيب جسيمات الفضة النانوية بالطرق الخضراء بواسطة النباتات شائعًا بشكل متزايد بسبب ملاءمته للبيئة ، وتوفره ، وفعاليته وكلفته القليلة ، ويمكن التعامل معه بأمان ويمتلك تنوعًا واسعًا من المركبات الفعالة ، مثل مضادات الأكسدة و ومركبات مضادة للميكروبات. في الدراسة الحالية ، تم تصنيع جسيمات الفضة النانوية AgNPs باستخدام المستخلصات المائية والميثانولية لأوراق الشاي الاخضر التي اختزلت نترات الفضة, والتي تم توصيفها باستخدام العديد من التقنيات التحليلية مثل القياس الطيفي المرئي فوق البنفسجي (UV-Vis) ، والتحويل الطيفي بالأشعة تحت الحمراء (FTIR) ، والفحص المجهري الذري (AFM) ، وتشتت الأشعة السينية (XRD) وفحص جهد زيتا. أشارت الملاحظة البصرية إلى أن لون نترات الفضة المائية قد تحول بعد المعاملة بمستخلصات الأوراق وتأكد ذلك بواسطة أطياف UV-Vis. فضلا عن ذلك ، أظهر تحليل AFM أن الجسيمات كانت كروية الشكل ، مفردة أو في مجاميع بمتوسط ​​أحجام 108.3 و 84.76 نانومتر للمستخلصات المائية والميثانولية على التوالي. كذلك, أكدت تقنية XRD الطبيعة البلورية للجسيمات النانوية, وتم تقدير متوسط ​​الحجم وفقًا لمعادلة شيرير اذ بلغت 61.24 و 99.66 نانومتر للمستخلصات المائية والميثانولية (CANPs) على التوالي. بالإضافة إلى قيم زيتا المحتملة كانت-30.31 و -32.33 مللي فولت للمستخلصات المائية والميثانولية CANPs على التوالي.

Synthesis, Characterization, and Photocatalytic Activity of Cu-Doped MgO Nanoparticles on Degradation of Methyl Orange (MO)

The purpose of this study is to synthesize Cu-doped MgO nanoparticles and test the performance of photocatalytic degradation of methyl orange (MO). Mg(NO3)2, CuCl2, NaOH, and fresh Calotropis procera leaf extract were used as precursors. The prepared nanoparticles were characterized by using FT-IR, XRD, SEM, and UV-Vis spectrometer to study the functional group, crystal structure, surface morphology, and absorption edge, respectively. The wide band above 3000 cm−1 from the FT-IR spectrum corresponds to the stretching vibrations of flavonoids and phenolic compounds of Calotropis procera leaf extract. Furthermore, the Mg-O bonding of undoped MgO and Cu-doped MgO NPs is represented by new peaks which appeared at 831 and 835 cm−1, respectively. The crystal size of undoped MgO and Cu-doped MgO nanoparticles is 13.04 nm and 12.08 nm, respectively. The SEM microstructure of pure MgO showed higher agglomeration than the Cu-doped MgO nanoparticles. The degradation efficiency of the Cu-doped MgO NPs was compared with that of the MgO NPs, and the photocatalytic activity of these NPs was evaluated using the photocatalytic degradation rates of MO dye. Cu-doped MgO NPs showed higher degradation efficiency than pure MgO NPs. The insertion of Cu in the MgO structure improved the photocatalytic efficiency of the MgO NPs under optimal conditions. Therefore, Cu-doped MgO exhibits high photocatalytic activity compared with undoped MgO nanoparticles under sunlight irradiation.

The Effect of Microwave-Assisted Extraction Temperature and Material to Solvent Ratio on the Characteristics of Pandan (Pandanus amaryllifolius Roxb.) Leaf Extract

Fresh pandan leaf fragrance has been applied to enhance the aroma of many traditional foods. However, the fresh leaf has a limited shelf life—only 1 day after harvest. Therefore, in liquid or powdered form, pandan extract can be utilized as an alternative aromatic component in food production. The purpose of this study was to determine the effect of extraction temperatures (50, 60, and 70°C) utilizing microwave-assisted extraction (MAE) and the ratio of pandan leaf as material to ethanol as solvent (1:3, 1:7, and 1:11) on the characteristics of pandan extract in liquid form. The study found that the properties of pandan extract treated at MAE 60°C with a material-solvent ratio of 1:7 were the best among all treatments. The pH of this liquid pandan extract was 5.31, the antioxidant activity was 32.86 ppm, and the chlorophyll content was 7.76 mg/L. The powdered pandan extract generated by drying the liquid pandan extract using a spray dryer and adding maltodextrin had properties such as water activity 0.45, solubility 96.09%, solubility rate 0.01 g/s, color L 85.65, a* -6.68, b* 8.51, yield 3.88%, and was detected to contain 0.17% of the aromatic compound 2-acetyl-1-pyrroline. Powdered pandan extract has the potential to be developed as an aromatic food additive. Panelists liked the Lumpang cakes, a traditional cake consisting of rice flour and coconut milk made by adding 1% pandan leaf extract powder.

Chemical constituents, antimicrobial and antioxidant activities of essential oils from fresh and air-dried leaves of <i>Icacina trichantha</i> (OLIV.)

We investigated and compared the constituents, antimicrobial, and antioxidant activities of the essential oils from the fresh and air- dried leaves of Icacina trichantha. The essential oils were extracted by hydrodistillation, antimicrobial activities were tested on Staphylococcus aureus, Salmonella typhi, Escherichia coli, Pleisomonas shigellosis, Bacillus cereus, Proteus vulgaris, Candida albicans, and Candida tropicalis by agar diffusion method, while 2,2- diphenyl-1-picrylhydrazyl (DPPH) was used for the antioxidant activities at 6.25-200 μg/mL of the oils compared with ascorbic acid as standard. The fresh and air-dried leaf oils gave 0.39% (w/w) and 0.33% (w/w) corresponding to 17 compounds representing 99.99%, and 10 compounds representing 100% respectively. Major compounds in the fresh leaf oil were 9-oxabicyclo [6.1.0] nonane (15.52%), Methyl isododecanoate(11.91%), Oleic acid (10.51%), 3-Cyclohexen-1-ol,3-methyl-(8.24%) and Phytol (7.70%), while Linoleic acid- (34.61%), Palmitic acid (20.74%), 9,17-Octadecadienal, (Z)- (12.98%), 9-Octadecenal (Z) (7.96%) and gamma-sitosterol (7.13%) were the major constituents of the air-dried oil. The fresh leaf oil 50 mg/mL, gave the highest zone of inhibition of 12 mm against Candida albicans while the air-dried oil had the highest zone of inhibition of 18 mm with a minimum inhibitory concentration of 0.20 mg/mL against C. tropicalis. The fresh leaf oil had the highest antioxidant activity of 73.85% at 100 μg/mL, while the air-dried and ascorbic acid had 70.25% and 69.23% respectively at 200 μg/mL. The fresh leaf oil comprised mainly oxygenated hydrocarbons (52.51%) while the air-dried oil had fatty acids (58.44%). Both oils were more antifungal than antibacterial and had a comparable antioxidant activity with ascorbic acid.

Reconstructing fresh green leaf spectra in the SWIR-2 region (2001–2500 nm) collected in a humid environment by referring to publicly available green leaf spectral databases

Leaf spectra (reflectance and transmittance) are commonly measured using a portable spectroradiometer and an integrating sphere or contact probe with an artificial light source. However, spectral data may be obscured due to water vapor and low signal-to-noise ratios, especially in the shortwave infrared-2 region (SWIR-2, 2001–2500 nm). Therefore, we proposed a spectral reconstruction approach to retrieve noise-free SWIR-2 fresh green leaf spectra by referring to the previously published quality-controlled fresh green leaf spectral reference databases. We processed 896 pairs of fresh tea (Camellia sinensis var. sinensis) leaf reflectance/transmittance from Alishan in central Taiwan. We selected a subset (500–1900 nm) of the spectra in the visible, near-infrared, and SWIR-1 regions (VNS-1) that were relatively insensitive to atmospheric conditions. We matched those spectra with publicly available reference green leaf spectral databases, and selected the one that was most similar to each Alishan VNS-1 spectrum. We then used multivariable linear regression, linear parameter multiplication and spectral reversion to reconstruct SWIR-2 spectra. Finally, we used another set of green leaf spectral databases to assess the performance of the proposed method. The performance of the reconstruction approach was satisfactory, with mean (± standard deviation) root-mean-square errors (RMSEs) of 0.0041 ± 0.0019 (reflectance) and 0.0054 ± 0.0027 (transmittance) for each spectrum and RMSEs of 0.0058 ± 0.0027 (reflectance) and 0.0055 ± 0.0043 (transmittance) for each SWIR-2 band. The proposed approach successfully modeled SWIR-2, which could be further improved with the availability of a more comprehensive set of green leaf reference spectral databases.

Recent advances in the genetic parameters and prospects of cultivation of the climate-smart herb senna (Senna alexandrina Mill.): A significant industrial medicinal crop

The bulk of senna plant components are used to make herbal medications to treat infections and other ailments. Except for grading, more than 80% of dried leaves are exported from India. The current study examined the genetic variability, correlations, and character contributions of 25 senna accessions. The amplitude of the GCV was larger than that of the PCV, showing that the selected factors interact genetically. X1 had substantial and positive connections with X4, X5, X6, X8, X11, and X13, while X2 had significant and positive links with X7, X10, X11, X12, X13, and X14. The characteristic X11 made the largest direct contribution to the X14 attribute. In X1 and X13, the indirect effect was greater. Fresh and dry leaf weight had high heritability and medium GA over the mean, respectively, whereas pod weight had high heritability and GA over the mean, at 93.62 and 94.93%. The mean, heritability, and GA over the mean of genotypes Gen 1, 2, 3, 4, and 25 were selected for high yield in the most economically significant traits. As a result, the senna genotypes mentioned above may be effectively exploited.

Investigation of morpho-physiolgical traits and gene expression in barley under nitrogen deficiency

Nitrogen (N) is an essential element for plant growth, and its deficiency influences plants at several physiological and gene expression levels. Barley (Hordeum vulgare) is one of the most important food grains from the Poaceae family and one of the most important staple food crops. However, the seed yield is limited by a number of stresses, the most important of which is the insufficient use of N. Thus, there is a need to develop N-use effective cultivars. In this study, comparative physiological and molecular analyses were performed using leaf and root tissues from 10 locally grown barley cultivars. The expression levels of nitrate transporters, HvNRT2 genes, were analyzed in the leaf and root tissues of N-deficient (ND) treatments of barley cultivars after 7 and 14 days following ND treatment as compared to the normal condition. Based on the correlation between the traits, root length (RL) had a positive and highly significant correlation with fresh leaf weight (FLW) and ascorbate peroxidase (APX) concentration in roots, indicating a direct root and leaf relationship with the plant development under ND. From the physiological aspects, ND enhanced carotenoids, chlorophylls a/b (Chla/b), total chlorophyll (TCH), leaf antioxidant enzymes such as ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT), and root antioxidant enzymes (APX and POD) in the Sahra cultivar. The expression levels of HvNRT2.1, HvNRT2.2, and HvNRT2.4 genes were up-regulated under ND conditions. For the morphological traits, ND maintained root dry weight among the cultivars, except for Sahra. Among the studied cultivars, Sahra responded well to ND stress, making it a suitable candidate for barely improvement programs. These findings may help to better understand the mechanism of ND tolerance and thus lead to the development of cultivars with improved nitrogen use efficiency (NUE) in barley.

Chromatography affinity resin with photosynthetically-sourced protein A ligand

Green, photosynthesizing plants can be proficiently used as cost-effective, single-use, fully biodegradable bioreactors for environmentally-friendly production of a variety of valuable recombinant proteins. Being near-infinitely scalable and most energy-efficient in generating biomass, plants represent profoundly valid alternatives to conventionally used stationary fermenters. To validate this, we produced a plastome-engineered tobacco bioreactor line expressing a recombinant variant of the protein A from Staphylococcus aureus, an affinity ligand widely useful in antibody purification processes, reaching accumulation levels up to ~ 250 mg per 1 kg of fresh leaf biomass. Chromatography resin manufactured from photosynthetically-sourced recombinant protein A ligand conjugated to agarose beads demonstrated the innate pH-driven ability to bind and elute IgG-type antibodies and allowed one-step efficient purification of functional monoclonal antibodies from the supernatants of the producing hybridomas. The results of this study emphasize the versatility of plant-based recombinant protein production and illustrate its vast potential in reducing the cost of diverse biotechnological applications, particularly the downstream processing and purification of monoclonal antibodies.

Effects of Neonicotinoid Seed Treatments on Cotton Seedling Physiology, Nutrition, and Growth

Plant growth and physiology can be affected by environmental and chemical factors that have the potential to influence yields. Among the factors that influence plant growth, neonicotinoid seed treatments have shown significant effects on plant growth, particularly in cotton. The dual benefits seen from neonicotinoids on plant growth and insect control show promise in improving cotton yields but little is known about how different seed treatments affect seedling physiology. A greenhouse experiment was undertaken to investigate how three neonicotinoid seed treatments (clothianidin, thiamethoxam, and imidacloprid) affect the physiology and growth of cotton seedlings in controlled environmental conditions. A randomized complete block design was used to examine seed treatments and an untreated control. Cotton seeds were treated, grown, and evaluated for physiological changes until the fifth true leaf-stage and measurements were taken at each of these stages. Data were collected on plant height, shoot fresh weight, leaf area, root length, and root biomass. In addition, chlorophyll pigments and nutrient analysis were performed on cotton seedlings. The seedlings of imidacloprid treated seeds had greater height, shoot fresh mass, leaf area, and relative growth rate by the fifth true leaf stage compared to other treated plants; however, clothianidin showed comparative performance at earlier stages in plant development that equilibrated over time. While all neonicotinoid seed treatments showed positive effects, imidacloprid showed the most potential as a bioactivator on plant growth.

IMPACTS OF WATER STRESS ON Urochloa brizantha FORAGE CULTIVARS

Water deficit during the dry season is one of the main factors affecting the productivity and quality of pastures. The present work aims at evaluating the effects of water deficit on the morphological and productive characteristics of Urochloa brizantha cultivars grown in Quartzarenic Neosol. The experiment was conducted in a greenhouse under a completely randomized design with a 4×2 factorial arrangement, with four cultivars (Marandu, MG4, MG13 Braúna and MG5 Vitória), two water availability conditions (50% or 100% of field capacity) and six repetitions. Structural, morphogenic and productive variables were evaluated 15, 25 and 35 days after the beginning of treatment application. For all productive characteristics evaluated during 35 days of water stress, it was observed that a 50% reduction in water availability significantly (P<0.05) reduced total fresh and dry mass production and leaf production of Marandu, MG13 Braúna and MG5 Vitória. Except for MG13 Braúna, the leaf/stem ratio of these cultivars was impaired. Regardless of water availability, when comparing forages during 35 days, lower cumulative leaf length and higher fresh and dry mass production of stems were observed in the Marandu cultivar. Regarding the response of productive characteristics, the MG4 cultivar showed the best results under water deficit when grown in Quartzarenic Neosol.

Does local soil factor drive functional leaf trait variation? A test on Neilingding Island, South China.

Leaf traits were affected by soil factors and displayed varietal differences in forest. However, few examples have been reported on the Island ecosystems. We comprehensively investigated 9 leaf traits (leaf length, leaf width, leaf area, SLA, leaf fresh weight, leaf C content, leaf N content, leaf K content, leaf C:N ratio) of 54 main subtropical woody species and soil parameters (soil pH, total C content, total N content, total K content, available N content, available P content, available K content and soil moisture) in Neilingding Island, Shenzhen, southern China. Intra-and interspecific variation of leaf traits were measured and their correlations with soil parameters were explored. The interspecific variations of leaf C:N ratio, leaf N content and leaf fresh weight were higher than their intraspecific variations. The intraspecific variation of leaf K content was larger than that of interspecific one, accounting for 80.69% of the total variance. Positive correlations were found among intraspecific coefficients of variations in leaf morphological traits. The correlation analysis between the variation of intraspecific traits and the variation of soil parameters showed that changes in soil factors affected leaf morphology and stoichiometry. The interaction between soil moisture and soil available P content was the key factor on intraspecific variations of leaf traits including leaf area, leaf fresh weight, leaf C and leaf K content. We concluded that leaf traits of plants in the island were tightly related to soil parameters. Soil parameters, especially soil moisture and available P content, affected plant leaf morphology and stoichiometry at the local scale.

Effect of Bacillus pumilus Strains on Heavy Metal Accumulation in Lettuce Grown on Contaminated Soil

Increasing number of heavy metal on land needs to be addressed through sustainable ways and various species of Bacillus can be used to mitigate heavy metals. The research work entitled “Effect of Bacillus pumilus strains on heavy metal accumulation in lettuce grown on contaminated soil” focuses on functional role of Bacillus pumilus strains inoculated with lettuce seed in mitigating heavy metal present in chromite mining soil. This experiment was conducted at ornamental horticulture nursery, the University of Agriculture Peshawar. In this experiment, factor A was three Bacillus pumilus strains (sequence C-2PMW-8, C-1 SSK-8 and C-1 PWK-7) while soil used for this experiment was collected from Prang Ghar mining site and lettuce seeds were grown in three levels of chromite mining soil (2.27, 4.65 and 7.14 %).The experimental design used during this research was randomized complete block design with two factors and was replicated thrice. The collected data related to effect of chromite mining soil and Bacillus pumilus strains indicated that these both factors have significant influence on growth parameters. For mining soil minimum days to germinate (14 days) was noted in lettuce grown on garden soil inoculated with sequence. Maximum germination percentage noted was (100%) for C-1 SSK-8 grown on garden soil, dry weight of lettuce leaf (5.0 g) for lettuce inoculated with C-1 SSK-8 and C-1 PWK-7 strains, survival percentage (77.9%) for C-1 SSK-8 treated lettuce for sequence C-2 PMW-8. Maximum germination percentage noted was (88%) for garden soil, fresh leaf weight (17.3g), dry leaf weight (6.5g), number of leaf per plant (16), leaf area (49.0 cm2) were noted in garden soil with no chromite mining soil. Results related to heavy metals accumulation showed that minimum chromium was (2.3 mg kg-1) in lettuce and (1.9 mg kg-1) in soil for all three sequences. It can be concluded that chromite mining soil significantly reduced the growth and survival of lettuce, but when lettuce was inoculated with Bacillus.pumilus strains it enhances the growth and survival. Similarly, minimum heavy metal accumulation in plant and soil, regardless of type of B. pumilus used, all three sequences has same mitigating effect on heavy metal in both soil and lettuce. All the three Bacillus pumilus strains ensured reduction in heavy metals content (Cr) in lettuce, below the maximum permissible limits of WHO/FAO 2011.

Citrus limon Wastes from Part of the Eastern Cape Province in South Africa: Medicinal, Sustainable Agricultural, and Bio-Resource Potential.

The fruits of Citrus limon are often purchased for their vitamin C-rich juice, while the fruit peel and the tree leaves are discarded as wastes. This study obtained the chemical profiles of the essential oils (EOs) of C. limon wastes (the peel and leaves), evaluated their medicinal value as antioxidants, their potential for sustainable use in agriculture as an insecticide for post-harvest preservation of grains, and their potential as a bioresource in livestock feed formulations. The EOs were isolated from C. limon leaves and peel using a hydro-distillation method on a Clevenger apparatus. The oil constituents were identified using the gas chromatography-mass spectrometry (GC-MS) hyphenated technique. The oils were evaluated for their in vitro antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing antioxidant power methods. An insecticidal study was conducted using contact toxicity, fumigation, and repellence bioassay methods against Sitophilus zeamais (maize weevils). Finally, the predicted income from using lemon peel as an alternative or substitute ingredient for maize in livestock feed formulations was obtained through a conventional simulation method. Chemically, limonene was found to be present in all the EOs analyzed (12-52%), while α-pinene was only found in the fresh leaf and peel oils (13.3% and 10.6%). Caryophyllene oxide was identified as the major component of the dried leaf oil (17.7%). At 20 µg m, the dry peel oil exhibited the highest inhibitory activity (52.41 ± 0.26%) against the DPPH radical, which was comparable to L-ascorbic acid (a standard antioxidant) at 54.25 ± 3.55%. The insecticidal study revealed that the dry peel oil is a better insect repellent (73.33 ± 6.95% at 10 µL) and fumigant (LC50 = 0.17 µL g-1 after 48 h) natural agent compared to the peel oil. Conversely, the dry peel oil showed a better contact activity (LC50 = 1.69 µL g-1) against the maize weevils compared to the dry leaf oil. The simulation study showed the cost of using dry lemon peel as an alternative to maize in livestock feed formulation to be ZAR 2.8 billion, compared against the higher cost of feed formulation with maize, which currently stands at ZAR 24.9 billion. This study has shown that C. limon wastes (the peel and leaves) contain EOs with unique chemical profiles, valuable medicinal properties as free radical scavengers, and considerable insecticidal properties for agricultural use in post-harvest grain preservation, presenting a cost-effective and promising bioresource for livestock feed production.

Efficient Expression of Functionally Active Aflibercept with Designed N-glycans.

Aflibercept is a therapeutic recombinant fusion protein comprising extracellular domains of human vascular endothelial growth factor receptors (VEGFRs) and IgG1-Fc. It is a highly glycosylated protein with five N-glycosylation sites that might impact it structurally and/or functionally. Aflibercept is produced in mammalian cells and exhibits large glycan heterogeneity, which hampers glycan-associated investigations. Here, we report the expression of aflibercept in a plant-based system with targeted N-glycosylation profiles. Nicotiana benthamiana-based glycoengineering resulted in the production of aflibercept variants carrying designed carbohydrates, namely, N-glycans with terminal GlcNAc and sialic acid residues, herein referred to as AFLIGnGn and AFLISia, respectively. Both variants were transiently expressed in unusually high amounts (2 g/kg fresh leaf material) in leaves and properly assembled to dimers. Mass spectrometric site-specific glycosylation analyses of purified aflibercept showed the presence of two to four glycoforms in a consistent manner. We also demonstrate incomplete occupancy of some glycosites. Both AFLIGnGn and AFLISia displayed similar binding potency to VEGF165, with a tendency of lower binding to variants with increased sialylation. Collectively, we show the expression of functionally active aflibercept in significant amounts with controlled glycosylation. The results provide the basis for further studies in order to generate optimized products in the best-case scenario.

Phenotypic and yield responses of common bean (Phaseolus vulgaris l.) varieties to different soil moisture levels

BackgroundMorphological plasticity is one of the capacities of plants to modify their morphological appearance in response to external stimuli. A plant’s morphology and physiology are constantly tuned to its variable surroundings by complex interactions between environmental stimuli and internal signals.In most of plant species,, such phenotypic and physiological expression varies among different varieties based on their levels of particular environmental stress conditions. However, the morphological and yield responses of common bean varieties to different environmental conditions are not well known. The purpose of the study was to evaluate morphological and yield response of common bean to soil moisture stress and to investigate the morphological mechanism by which common bean varieties tolerate fluctuations in moisture stress.MethodsA pot experiment was carried out to investigate the effects of different moisture levels on the phenotypic and yield responses of common bean varieties. A factorial combination of five common bean varieties (Hirna, kufanzik, Awash-1, Ado, and Chercher) and three moisture levels (control, waterlogging stress, and moisture deficit stress) was used in three replications. Moisture stress treatments were started 20 days after planting, at the trifoliate growth stage. To evaluate the response of each variety, morphological and yield data were collected at week intervals.Main resultsThe results indicated that moisture levels and varieties had a significant influence on all growth parameters. Crop phenology was significantly influenced by the interaction effect of moisture level and variety. Exposing Hirna variety to moisture stress led to extended flowering and pod setting by 23 and 24 days, respectively, compared to the other treatments. The results showed that the phenotypic responses to moisture deficit and waterlogging stress varied between varieties. Waterlogging stress had a stronger reduction effect on the fresh weight, dry weight and leaf area of common bean varieties than moisture deficit and the control. Pods per plant, seeds per plant, grain yield per plant, and harvest index were significantly influenced by the varieties, moisture stress levels and their interaction. Except for Chercher and Hirna. However, varieties Ado, kufanzik and Awasha-1 did not show significant differences on the time of flower initiation due to moisture level. Biomass and growth in leaf fresh weight, leaf dry weight, leaf area, leaf number and plant height were significantly influenced by moisture level. When moisture deficit and waterlogging stress occurred, Ado and Awash-1 were more responsive to moisture stress than Hirna, Chercher, and Kufanzik.ConclusionHence, Hirna and Kufanzik varieties were found to be tolerant because they produced higher yields than the Chercher, Awash-1, and Ado varieties.

Adding value to banana farming: Antibody production in post-harvest leaves

Banana, a globally popular fruit, is widely cultivated in tropical and sub-tropical regions. After fruit harvest, remaining banana plant materials are low-value byproducts, mostly composted or used as fibre or for food packaging. As an aim to potentially increase farmer income, this study explored underutilised banana biomass as a novel plant tissue for production of a high-value product. Protein scFvTG130 used in this study, is an anti-toxoplasma single chain variable fragment antibody that can be used in diagnostics and neutralising the Toxoplasma gondii pathogen. Using detached banana leaves, we investigated the factors influencing the efficacy of a transient expression system using reporter genes and recombinant protein, scFvTG130. Transient expression was optimal at 2 days after detached banana leaves were vacuum infiltrated at 0.08 MPa vacuum pressure for a duration of 3 min with 0.01% (v/v) Tween20 using Agrobacterium strain GV3101 harbouring disarmed virus-based vector pIR-GFPscFvTG130. The highest concentration of anti-toxoplasma scFvTG130 antibody obtained using detached banana leaves was 22.8 µg/g fresh leaf tissue. This first study using detached banana leaf tissue for the transient expression of a recombinant protein, successfully demonstrated anti-toxoplasma scFvTG130 antibody expression, supporting the potential application for other related proteins using an underutilised detached banana leaf tissue.

Analysis of tea quality of large-leaf black tea with different harvesting tenderness based on metabolomics

The harvesting tenderness of tea fresh leaves has a significant effect on the flavor and aroma quality of black tea (BT), which is one of the important standards for BT grading. In this study, sensory evaluation, electronic tongue technique and metabolomics were performed to evaluate the flavor and aroma qualities of three different tenderness of large-leaf BTs that were manufactured from the fresh leaves of single-bud, one-bud and one-bud two-leaf teas, respectively. The results of the electronic tongue and sensory evaluation indicated that the increase in the tenderness of the fresh leaf material enhanced the sweetness, freshness and fruity flavor of the tea and reduced the bitterness of the tea. A total of 519 differential flavor components and 84 differential aroma components were identified by metabolomics. Among them, elevated levels of maltitol, cyclic leucine and l-methionine and l-aspartic acid led to elevated sweetness and freshness in more tender tea leaves. Reduced levels of 4-aminosalicylic acid, tyrosol, uracil, inosine, benzamide, and l-tyrosine led to reduced bitterness in BT. Moreover, the content of volatiles such as (Z)-2-Decenal, Phenylacetic acid propyl ester, Acetophenone, and 2-phenylethyl ester were increased, which contributed to the fruity aroma in the high-tenderness tea. This study provides a theoretical basis for grading the flavor quality of BT with different tenderness.