Fresh Leaf Research Articles

Application of ATR-FTIR Spectroscopy for Analysis of Salt Stress in Brussels Sprouts.

Salt stress is one of the environmental stresses that significantly reduces crop productivity and quality worldwide. Methods to overcome salt stress include developing salt-resistant crops by inserting various resistance genes or to diagnosing and responding to the effects of salt stress at an early stage. In this study, we investigate the effects of salinity stress on growth, photosynthetic efficiency, and metabolic changes in Brussels sprouts (Brassica oleracea var. gemmifera). Fresh weight and leaf area decreased significantly with increasing NaCl concentration, indicating that salinity stress has a detrimental effect on plant growth. However, chlorophyll fluorescence parameters did not show significant changes, suggesting that photosynthetic efficiency was not significantly affected over 10 days. Fourier transform infrared (FTIR) spectroscopy revealed notable metabolic adjustments, especially in lipids, plastids, proteins, and carbohydrates, indicating biosynthesis of protective compounds such as anthocyanins and proline in response to salinity stress. Pearson correlation analysis confirmed a strong relationship between NaCl concentration and the observed physiological and metabolic changes. The findings highlight the potential of FTIR spectroscopy as a non-destructive tool for early detection of salinity stress and timely intervention to improve crop resilience and yield. This study highlights the widespread application of FTIR spectroscopy in agricultural research to manage abiotic stresses in crops.

Evaluating Seawater and Wood Distillate for Sustainable Hydroponic Cultivation: Implications for Crop Growth and Nutritional Quality

The adoption of innovative cultivation methods, such as hydroponics and aeroponics, is gaining attention due to the unprecedented demand for food that an increasing population is posing on agricultural systems, exacerbating the pressure on already limited arable land. Seeking sustainable and circular economy solutions is imperative, aiming to optimize water consumption and enhance crop yields and quality without resorting to synthetic chemical fertilizers. This study investigated the use of seawater at various concentrations as a base for nutrient solutions, with and without the addition of a natural biostimulant, wood distillate (WD). Four seawater (SW) concentrations (0, 3, 6, and 12%) and two wood distillate concentrations (0 and 0.2%) were applied to assess their impacts on lettuce growth. Findings reveal that seawater at low concentrations (< 6%) serves as an effective water-saving strategy, despite the reduction in the plant ascorbic acid contents. The addition of WD did not inflate growth; in fact, the results obtained are comparable to that of the controls for each concentration of seawater, except at the highest concentration (12% SW), resulting in reduced fresh leaf weights and root areas. Significantly, there was a notable increase in the ascorbic acid contents in all plants grown with WD. Moreover, the WD increased the leaf concentrations in Ca, Mg, P, and K, indicating the higher nutritional value of the crop. This research highlights the potential of combining seawater and WD for sustainable and efficient plant cultivation, suggesting new strategies for exploration across diverse plant species and hydroponic applications.

Formulation and characterization of natural deep eutectic solvents (NADES) for simultaneous phenolics and carotenes extraction from fresh oil palm leaf

In this study, natural deep eutectic solvents (NADES), comprising choline chloride (ChCl) as hydrogen bond acceptor (HBA) with hydrogen bond donor (HBD) including glucose (Glu), fructose (Fru), xylose (Xyl), xylitol (Xyli), and glycerol (Gly), were formulated and characterized for simultaneous extraction of phenolic compounds (polar) and carotenoids (non-polar) from fresh oil palm leaves (OPL). pH, density, polarity, Fourier Transform Infrared Spectroscopy (FT-IR) and rheological analyses were conducted, with relation to the extraction efficiency achieved via homogenizer-assisted heating and stirring extraction approach. Among the NADES formulations tested, ChCl:Xyl (1:1) displayed the highest total phenolic content (TPC) at 20.23825 mg/g OPL, due to its optimal polarity at 49.4654 kcal mol−1 and a viscosity of 23288 mPa.s. The highest total carotenoid content (TCC) was observed in ChCl:Xyli (1:1) at 325.94 µg/g OPL, with a lower polarity at 50.0280 kcal mol−1. Despite having similar viscosity and polarity, ChCl:Fru (1:2) with a pH of 5.82 outperformed ChCl:Glu (1:1) with a pH of 6.72 due to enhanced phenolic compound solubility and stability. This study shed light on the potential of NADES, particularly in overcoming the challenge of high moisture content in OPL and other fresh biomasses, which often acted as a barrier during phytonutrients extraction.

Mycorrhizal symbiosis and natural dye waste organic fertilizer: Enhancing growth and yield in indigofera tinctoria

Background: Indigofera tinctoria, commonly known as true indigo, is a plant widely used in the textile industry for its natural indigo dye, which produces a rich blue color for fabrics. Indigofera tinctoria, known for its natural bluish-purple dye, generates 10% dye and 90% waste during extraction, requiring effective waste management. This study aimed to optimize organic fertilizer and mycorrhiza doses from I. tinctoria waste for enhancing soil fertility in arid regions. Methods: The study was conducted from May to December 2020 in Puron Village, Bulu District, Sukoharjo Regency. A factorial Completely Randomized Block Design (CRBD) with 2 factors was employed. The first factor consisted of 5 levels of I. tinctoria organic fertilizer treatments: 0, 100, 200, 300, and 400 g per plant. The second factor included 3 levels of mycorrhiza treatments: 0, 10, and 20 g per plant. Variables observed included mycorrhizal infection on roots, plant growth rate, and yield (fresh leaf and shoot weight of I. tinctoria). Data analysis utilized ANOVA at a 5% significance level followed by Duncan Multiple Range Test (DMRT). Results: The results indicated that organic fertilizer from natural dye waste at a dosage of 200 g per plant increased the percentage of mycorrhizal-infected roots and boosted shoot weight by 63.27% at 8 weeks after planting (WAP). Mycorrhiza at 10 g per plant increased mycorrhizal infection percentage and enhanced shoot weight by 45.98% at 4 WAP. The combination of I. tinctoria extraction waste organic fertilizer at 200 g per plant and mycorrhiza at 10 g per plantshowed interaction, significantly increasing the growth of root nodules of Indigofera tinctoria by 84.04% at 12 WAP. Conclusion: The integration of organic fertilizer derived from indigo dye waste and mycorrhiza presents a promising strategy for enhancing I. tinctoria growth and productivity. Novelty/Originality of this Study: This study is distinctive in its demonstration of the effective use of I. tinctoria extraction waste as an organic fertilizer, aligning with zero-waste principles and contributing to improvements in plant growth and soil fertility. Furthermore, it investigates the synergistic effects of mycorrhizal associations on enhancing nutrient absorption and overall productivity of I. tinctoria, an aspect that has not been thoroughly explored in prior research.

First Report of 'Candidatus Phytoplasma ziziphi'-Related Strains Infecting Peony (Paeonia suffruticosa Andr.) Showing Leaf Yellows Symptoms in China.

Peony (Paeonia suffruticosa Andr.), belonging to family Paeoniaceae, is an important medicinal and ornamental plant. During August of each year from 2016 to 2023, peony plants at Heze city were found to exhibit leaf yellows symptoms. The incidence rate of the symptomatic plant was recorded from 10% to 30% in four peony gardens with about 200 acres. Total DNA was extracted from 0.10 g fresh plant leaf tissues from 24 symptomatic and 8 asymptomatic samples using rapid plant genomic DNA isolation kit (Aidlab Biotechnology, Beijing, China). The extracted DNA was amplified by nested polymerase chain reaction using universal primers R16mF2/R16mR1 followed by R16F2/R16R2 (Lee et al., 1993; Gundersen and Lee, 1996) specific for the 16S rRNA gene and new designed tuf gene specific primers JWB-tuforfF1 (5'-ATGGCTGAAATATTTTCAAGAG-3') and JWB-tuforfR1 (5'-TTATTCTATGATTTTAATAACAG-3') followed by JWB-tuforfF2 (5'-ATGTAAACGTAGGAACTATTGG-3') and JWB-tuforfR2 (5'- TCCGATAGTTCTTCCACCTTCAC-3'). Amplicons of about 1.25 kb and 1.02 kb (16S rRNA gene and tuf gene, respectively) were obtained in 8 symptomatic samples from four peony gardens. However, no amplification was obtained in any of the asymptomatic samples. The representative amplicons of 16S rRNA and tuf genes of three positive samples (Heze-9, -18 and -27) were cloned into a zero background pLB-simple vector (Tiangen Biotechnology, Beijing, China) and sequenced by Taihe Biotechnology, Beijing, China. Sequences obtained in the study were deposited in NCBI GenBank with accession numbers PP504882, PP504883 and PP504884 for the 16S rRNA gene as well as PP530237, PP530238 and PP530239 for the tuf gene. The phytoplasma strain under the study was described as peony yellows (PeY) phytoplasma, PeY-Heze strain. Alignment analysis by DNAMAN software showed that three 16S rRNA gene sequences obtained in the study shared 99.36% to 99.60% sequence identity and three tuf gene sequences obtained in the study were identical. BLAST analysis of the 16S rRNA gene sequences of the PeY-Heze phytoplasma strains showed 99.60%-99.84% sequence identity with 'Candidatus Phytoplasma ziziphi' (GenBank accession: CP025121). And tuf sequences of the strains showed 100% similarity with 'Ca. P. ziziphi' (CP025121). Interestingly, the virtual RFLP patterns derived from three 16Sr RNA gene sequences obtained in the study by iPhyClassifier (Zhao et al., 2009) were different from the reference patterns of all previously established 16Sr groups/subgroups. The most similar are the reference pattern of the 16Sr group VII, subgroup E (AY741531), with a similarity coefficient of 0.72, which is less than 0.85. These phytoplasma strains may represent a new 16Sr group. Phylogenetic analysis based on 16S rRNA genes using MEGA 7.0 by neighbor-joining (NJ) method with 1000 bootstrap value indicated that PeY-Heze strains clustered into one clade with the phytoplasma strains of 'Ca. P. ziziphi' with 68% bootstrap value. Although there are several reports available on 'Ca. P. solani' infecting peony in Shandong Province, China (Gao et al., 2013). To our knowledge, this is the first report of 'Ca. P. ziziphi'-related strains infecting peony in China. The findings in this study will be beneficial to the detection, quarantine, and prevention of peony yellows phytoplasmas in China.

Eurochevalierines A-I, Sesquiterpene Alkaloid Hybrids with Anti-Triple Negative Breast Cancer Activity from Penicillium sp. HZ-5.

Nine new sesquiterpene alkaloids, eurochevalierines A-I (1-9), were separated from the rice cultures of the endophytic fungus Penicillium sp. HZ-5 originated from the fresh leaf of Hypericum wilsonii N. Robson. The structures' illumination was conducted by single-crystal X-ray diffraction, extensive spectroscopic analysis, alkaline hydrolysis reaction, and Snatzke's method. Importantly, the antitumor activities screen of these isolates indicated that 1 could suppress triple negative breast cancer (TNBC) cell proliferation and induce apoptosis, with an IC50 value of 5.4 μM, which is comparable to the positive control docetaxel (DXT). Flow cytometry experiments mentioned that compound 1 significantly reduced mitochondrial membrane potential (MMP) of TNBC cells. In addition, 1 could activate caspase-3 and elevated the levels of reactive oxygen species (ROS) and expressions of suppressive cytokines and chemokines. Further Western blot analysis showed that 1 could selectively induce mitochondria-dependent apoptosis in TNBC cells via the BAX/BCL-2 pathway. Remarkably, these finding provide a new natural product skeleton for the treatment of TNBC.

Fresh Moringa Stenopetala leaves consumption and its determinants among pregnant women in southern Ethiopia.

A woman's health and nutritional status has significant impact on her pregnancy situation. However, many pregnant women are undernourished. Moringa stenopetala is a plant consumed worldwide in various forms, and its consumption showed a reduction in the incidence of malnutrition. Although Moringa stenopetala is rich in essential macro- and micronutrients, there is little evidence on the proportion and determinants of fresh Moringa stenopetala leaf intake among pregnant women. The objective of this study was to fill this gap in the littérature and provide a baseline evidence for further research or intervention by investigation the proportion and determinants of fresh Moringa stenopetala leaf intake among pregnant women in the Gamo zone, south Ethiopian region. A community-based cross-sectional study was conducted among 623 randomly selected pregnant women using a pre-tested and structured questionnaire via a face-to-face interview. The consumption pattern was assessed based on a self-reported dietary history over the last 30 days before data collection. Multivariable logistic regression model was fitted using STATA version 14. An adjusted odds ratio with a 95% confidence interval was reported to show an association between the dependent and independent variables with level of statistical significance at a p-value of <0.05. The proportion of fresh Moringa stenopetala leaves intake among pregnant women was 49.60% (95% CI: 45.67, 53.52%). The déterminants of fresh moringa leaf intake were being below 24 years old (AOR: 2.92; 95% CI: 1.51, 5.63), rural résidence (AOR: 1.97; 95% CI: 1.10, 3.50), antenatal care attendance (AOR: 2.08; 95% CI: 1.03, 4.21), history of contraceptive use (AOR: 1.88; 95% CI: 1.03, 3.55), and having a good knowledge about the importance of moringa Stenopetala (AOR: 9.76; 95% CI: 5.30, 17.95). The study showed that almost half of the pregnant women consumed fresh Moringa stenopetala leaves. Women's age, place of residence, prenatal care, history of contraceptive use, and knowledge of the benefits of Moringa Stenopetala were positively associated with the consumption of fresh Moringa Stenopetala leaves. Therefore, health authorities and stakeholders involved in maternal and child health need to target older women, and urban residents and promote the benefits of consumption by strengthening uptake of maternal health services and raising awareness about Moringa Stenopetela. Future studies involving large scale and longitudinal designs evidence are required to further validate the findings so that this nutritious diet can be promoted widely among pregnant women in the study area and Ethiopia at large.

Investigating the crucial role of salicylic acid and harvest time on the yield and essential oil composition in sage (Salvia officinalis L.)

This research was carried out to determine the effects of different harvest time (HT) and salicylic acid application (SAA) on agronomic and quality characteristics of sage. The experiment was carried out in a randomized complete block design in split plots with HTs (pre-flowering and 50% flowering periods) in the main plots and SAAs in the subplots (Control, 1, 2, and 4 mM). Plant height, fresh herb yield, dry herb yield, fresh leaf yield, dry leaf yield, leaf ratio, essential oil (EO) content, EO yield, and EO composition were examined. In total or mean results, the HT was statistically significant on plant height, fresh and dry herb yield, leaf ratio in both years. The SAA was statistically significant on fresh and dry herb yield and leaf ratio only in the second year. Among the 24 components identified, alpha thujone was the main component. This component was followed by camphor, 13-epimanool, veridiflorol, 1.8-cineol. According to the principal component analysis (PCA), in 2021, 1 mM SAAs applied at the 50% flowering period in the first cutting and at the pre-flowering period in the second cutting showed different properties regarding EO composition. In 2022, 4 mM SAAs applied at the 50% flowering period in the first and second cuttings showed different properties regarding EO composition. The study found that delaying the HT led to significant increases in plant height, fresh and dry herb yields. Additionally, the SAA increased the herb yield and decreased the leaf ratio, particularly in the second year.

Sole-source Lighting of Lettuce that Increased Yield Had No Negative Effect on Postharvest Longevity

The effects of sole-source lighting on the growth and yield of hydroponically grown lettuce have been extensively studied, but research of postharvest performance is limited. We grew frill-leaf lettuce (Lactuca sativa) ‘Green Incised’ and ‘Hydroponic Green Sweet Crisp’ hydroponically in an indoor vertical research farm under daily light integrals (DLIs) of 12 or 18 mol⋅m−2⋅d−1 and the following three ratios of blue (B; 400–499 nm) and red (R; 600–699 nm) light from light-emitting diode fixtures: B5:R95, B20:R80, and B35:R65. We postulated that biomass accumulation would increase with the DLI and decrease with the B light fraction, and that postharvest longevity would increase with the DLI and the B light fraction. As expected, shoot fresh weight, leaf length and width, leaf number, and relative chlorophyll content (SPAD; ‘Green Incised’ only) decreased as the proportion of B light increased from 5% to 35%. Decreasing the DLI from 18 to 12 mol⋅m−2⋅d−1 reduced the shoot fresh weight and leaf number of both cultivars. Leaves of ‘Green Incised’ were up to 27% wider under B5:R95 and 60% longer under B5:R95 at 12 mol⋅m−2⋅d−1 than those under treatments with a higher DLI or more B light. The shoot fresh weight of ‘Hydroponic Green Sweet Crisp’ was greatest when grown under B5:R95 at 18 mol⋅m−2⋅d−1 and decreased as B light increased or DLI decreased. At the time of harvest, leaves of each cultivar and treatment were placed in clamshells and stored at 7 °C in darkness and evaluated for decay. ‘Green Incised’ that grew under B35:R65 and a DLI of 18 mol⋅m−2⋅d−1 had the shortest storage life, with 9.5 d and 11.4 d for replications 1 and 2, respectively, which were ∼2.5 to 4.0 d and 1.4 to 3.6 d earlier, respectively, than the storage life of lettuce grown under other treatments. In contrast, ‘Hydroponic Green Sweet Crisp’ was not influenced by light quality or DLI and had a storage life of 12.6 to 13.3 d and 13.5 to 14.3 d for replications 1 and 2, respectively. Therefore, a B light fraction between 5% and 20% and a DLI of 18 mol⋅m−2⋅d−1 produced high-yielding frill-leaf lettuce with a relatively long storage life.

Simulation and analysis of movement trajectories of fresh tea leaf particles based on CFD‐EDEM coupling

AbstractTo study the air‐suction separation mechanism of fresh tea leaves, this article adopts the coupling simulation method of discrete elements and computational fluid dynamics to conduct numerical simulation and experimental research on the movement of fresh tea leaves adsorbed by negative pressure air flow. In the gas–solid coupling model, three‐dimensional scanning technology is applied to establish a three‐dimensional model of fresh tea leaves with one bud and two leaves. Through the Fluent module in CFD, the negative pressure adsorption gas phase of the simulation process was established, bench tests were established, and parameters of the coupling model were optimized. Finally, the minimum error between the simulated value and the test value was 2.85% and the maximum error was 7.43%, indicating that the coupling model established had certain accuracy and could be theoretically analyzed according to the coupling model established. It is found that when the static pressure values of the adsorption surface are −120 Pa, −340 Pa, and −500 Pa respectively, the initial distance from the adsorption surface is 250–550 mm. It is found that fresh tea leaves with one bud and two leaves all fall to the pipe before adsorption, and then move to the adsorption surface. Under the same static pressure value of the adsorption surface, the initial falling distance of tea particles from the adsorption surface (250–550 mm) has a linear function relationship with the distance between the turning point and the adsorption surface. When the initial distance between falling tea particles and the adsorption surface is the same, the distance between the turning point and the adsorption surface is an exponential function when the initial position is 250–350 mm from the adsorption surface, and the distance between the turning point and the adsorption surface is a linear function when the initial position is 400–550 mm from the adsorption surface.

Response of Vegetative Growth and Yield of Two Hybrids of Broccoli (Brassica oleracea var. italica) to Liquid Organic Fertilizer and Hefe Algae

This research was conducted to evaluate the impact of liquid organic fertilizer and Hefe algae (seaweed) on various aspects of vegetative growth and yield in two hybrids of broccoli. The results revealed that the Agassi RZ hybrid outperformed the Wisdom hybrid in parameters such as leaf area (cm2), fresh leaf weight leaves (g), dry leaf weight (g), total yield (kg plant-1), and total yield (t ha-1). The foliar application of 3.0 and 6.0 ml L-1 of organic fertilizer significantly enhanced all vegetative growth and yield parameters. Similarly, Hefe algae, particularly at a concentration of 8.0 ml L-1, showed positive and significant effects on plant height (cm), number of leaves (leaves plant-1), leaf area (cm2), fresh leaf weight (g), dry leaf weight (g), chlorophyll%, total yield (kg plant-1), and total yield (t ha-1). The combined influence of two factors, namely hybrids and organic fertilizer, hybrids and Hefe algae, and organic fertilizer and Hefe algae, had a significant impact on all vegetative growth and yield parameters. Notably, the interaction among Agassi RZ hybrid, 6.0 ml L-1 organic fertilizer, and 8.0 ml L-1 Hefe algae exhibited the most substantial effects, resulting in the highest plant height (cm), number of leaves (leaves plant-1), fresh leaf weight (g), dry leaf weight (g), total yield (kg plant-1), and total yield (t ha-1).

DETERMINATION OF ANTIFUNGAL AND ANTIOXIDANT POTENTIAL OF LEAF AND FRUIT EXTRACTS OF GUAVA (Psidium guajava Linn.)

The present study was aimed to evaluate the antifungal and antioxidant properties of leaf and fruit extracts of guava (Psidium guajava Linn.). Antifungal assays were conducted to assess the efficacy of extracts against clinically relevant fungal strains, viz., Aspergillus niger MTCC-4325, Penicillium chrysogenum MTCC-2539, Candida albicans MTCC-7315, and Fusarium solani MTCC-3309. Besides, enzyme activity for catalase (CAT) and peroxidase (POD) were assayed to elucidate the antioxidant potential of extracts. The radical scavenging activities of extracts were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays to quantify the overall antioxidant capacity. Results indicated pronounced inhibitory activity against F. solani in methanolic extracts of leaves (21.66 ± 2.05) and fruits (24.66 ± 1.25), with aqueous extracts exhibiting comparatively lesser inhibition across the tested fungal strains. Moreover, the enzyme activity assays revealed higher CAT and POD activities in fresh leaf extract as compared to the fruit extract. Notably, DPPH assay underscored the superior antioxidant potential of Psidium leaf extracts relative to fruit extracts, while FRAP assay showed higher radical scavenging capacity of methanolic extract as compared to the aqueous extract.

Comparative analysis of red and green lettuce microgreens under different artificial LED lighting conditions

This study investigated how LED light exposure influences the growth and nutrient content of two lettuce microgreens over 10 days in a randomized setup. Lettuce seeds underwent surface sterilization, germination in prepared soil, and exposure to different LED light conditions. Morphological parameters and pigment analysis, including stem length, petiole length, leaf area, plant height, root length, fresh weight, dry matter percentage, were evaluated. Green lettuce outperformed red lettuce in the studied morphological parameters, including stem length (2.74±0.22 cm), plant height (4.54±0.21 cm), and fresh weight (3.79±0.32 g/100 plants) under different LED. White light promoted taller plants with higher fresh weight (4.45±0.43 g/100plants), dry matter (4.84±0.38%), and leaf area (0.76±0.06 cm2) in both lettuce species. In contrast, red light reduced overall growth and development, as evidenced by a 54% decrease in leaf area, despite a 23.36% increase in plant height. Chlorophyll levels varied significantly among LED treatments, with white LED yielding the highest levels in both red and green lettuce. Highest chlorophyll a (146.37±6.27 µg/g FW), chlorophyll b (86.74±2.44 µg/g FW), total chlorophyll (233.11±8.69 µg/g FW) and relative chlorophyll (215.84±8.05 µg/cm²) content was found in green lettuce under white light condition. Similarly, green lettuce grown under white LED had the highest total carotenoid, β-carotene, and lutein. The study concludes that optimizing white LED illumination has the potential to improve the nutritional value of lettuce microgreens by enhancing growth and pigment content, particularly in green varieties. These findings emphasize the crucial role of LED light color in optimizing the nutritional quality of microgreens.

Contribution of Aerobic Cellulolytic Gut Bacteria to Cellulose Digestion in Fifteen Coastal Grapsoid Crabs Underpins Potential for Mineralization of Mangrove Production

Grapsoid crabs (Decapoda: Grapsoidea) inhabiting along the land-sea transition provided various amounts and quality of vascular plant carbon (e.g., fresh mangrove leaf, leaf litter, and mangrove-derived organic carbon) and perform differing levels of herbivory. Other than endogenous cellulase, symbiotic cellulolytic bacteria could also contribute to the crabs’ vascular plant carbon assimilation and mineralization. In this study, we isolated culturable cellulolytic bacteria from three gut regions (i.e., stomach, midgut, and hindgut) of 15 species of grapsoid crabs that inhabit in various coastal habitats (i.e., land margin, mangrove forest, tidal flat, and subtidal area). Bacillus, which was isolated from 11 out of the 15 grapsoid crabs, was the most common genus of culturable prominently cellulolytic bacteria among the target species. Seventy to ninety nine percent of culturable cellulolytic bacteria were removed, and the endoglucanase activity of five species was significantly reduced by 14.4–27.7% after antibiotic treatment. These results suggest that cellulolytic bacteria play a role in assisting mangrove carbon utilization in coastal grapsoid crabs, especially those inhabiting mangrove, mudflat, and subtidal areas. The significantly higher abundance of cellulolytic bacteria and the generally higher hydrolytic capacity of the bacteria in mangrove crab species suggest that they receive more contribution from symbionts for mangrove carbon utilization, while semi-terrestrial crabs seem to depend little on symbiotic cellulase due to the lower abundances.

Physiological and Cellular Ultrastructural Responses of Isatis indigotica Fort. under Salt Stress

This study aimed to analyze the effects of salt stress on the growth physiology and plant-cell ultrastructure of Isatis indigotica Fort. (I. indigotica) to evaluate its adaptability under salt stress. The effects of different concentrations of salt (NaCl; 0, 25, and 300 mmol·L−1) on the agronomic traits, activities of related enzymes, ion balance, and mesophyll-cell ultrastructure of I. indigotica were studied in a controlled pot experiment. Results showed that compared with those of the control group, the aerial-part fresh weight, underground fresh weight, tiller number, root length, root diameter, plant height, and leaf area of salt-stressed I. indigotica increased at 25 mmol·L−1 and then decreased at 300 mmol·L−1. The changes in levels of superoxide dismutase, peroxidase, ascorbate peroxidase, and catalase showed a similar trend, with significant differences compared with control group. Salt stress altered the ion balance of I. indigotica, resulting in a significant increase in Na+ content and a significant decrease in K+ content. The contents of Ca2+ and Mg2+ changed to varying degrees. The analysis of the microstructure of the root showed that under salt treatment, the epidermal cells of the root significantly thickened and the diameter of the xylem decreased. The results of ultrastructural analysis of mesophylls showed that salt stress can cause cell-membrane contraction, cell-gap enlargement, disorder in the structures of chloroplasts and mitochondria, and an increase in the number of osmiophilic particles. These changes were aggravated by the increase in NaCl concentration. This study reveals the response of I. indigotica to salt stress and provides a basis for further study on the salt-tolerance mechanism of I. indigotica.

Productivity of elephant grass pakchong (Pennisetum purpureum cv Pakchong) and elephant grass taiwan (Pennisetum purpureum cv Taiwan) cultivated based on different stem cutting sizes

Abstract In Indonesia, elephant grass seeds used by farmers are in the form of cuttings or vegetative material. Currently, there is no standard for the cuttings used to produce high-productivity elephant grass. Therefore, research was conducted to determine the optimal size of cuttings for achieving good elephant grass productivity. The study was conducted in the field station area of Rancamaya, Bogor. A factorial group randomized design with 5 replications was employed for the research. The first factor was the type of elephant grass, consisting of 1) Pakchong and 2) Taiwan, while the second factor was the size of the cuttings, consisting of 1) 5 cm long cuttings, one node, 2) 10 cm long cuttings, one node, 3) 15 cm long cuttings, one node, and 4) control (two nodes as per usual farmer practice). The research results indicated that there was an interaction between the type of elephant grass and the size of cuttings for leaf length, fresh and dry leaf weight, dry stem weight, and total dry weight. The 5 cm cuttings of Pakchong had the highest fresh and dry leaf weights (P&lt;0.05). The highest dry stem and total weight were obtained with the control size cuttings of Pakchong. The number of tillers was influenced by the size of cuttings, with the control size cuttings having the highest number of tillers, although not significantly different from the number of tillers for the 5 cm and 15 cm cuttings. The highest total fresh weight was obtained with the 5 cm cuttings and Pakchong. This study indicates that 5 cm cuttings can be used as cutting material to produce Pakchong and Taiwan with optimal forage production. Further research is needed to assess the forage quality.

Selected Beneficial Microbes Alleviate Salinity Stress in Hydroponic Lettuce and Pak Choi

Hydroponics is widely used in greenhouse and vertical farming production because these facilities can precisely control environmental conditions such as lighting, temperature, and vapor pressure deficit. However, the fertilizer solutions have a short life span, and they often do not have adequate microbial populations to enhance plant growth. Previous studies have shown the potential of beneficial microbes to promote plant production and alleviate abiotic and biotic stressors in the field, and studies on their use in controlled environments such as greenhouses and vertical farms are limited in the literature. In this study, we selected several plant growth promoting microbes (PGPMs) and tested their effects on alleviating salinity stress in ‘Rex’ lettuce (Lactuca sativa) and ‘Red Pac’ pak choi (Brassica chinensis) grown in deep water culture hydroponics. Our goal was to use one stressor, salinity, that induces profound symptoms in plant morphology. A three-cycle study was conducted using five PGPMs [Bacillus, Glomus, Lactobacillus, Trichoderma, and Bacillus/Pseudomonas/Trichoderma (B/P/T) mix] and two salinity levels [no salinity and salinity treatment, with 120 mM, 40 mM, and 80 mM sodium chloride (NaCl) solution used for the first, second, and third cycles, respectively]. We measured the effects of PGPMs and salinity on plant growth and quality and the solution pH and electrical conductivity (EC). Salinity stress decreased lettuce and pak choi leaf area and shoot fresh weight and increased plant leaf chlorophyll and anthocyanin contents with increased solution EC. Under high-salinity stress (120 mM NaCl), the addition of Trichoderma reduced pak choi leaf area and fresh weight but increased solution pH, whereas under low salinity stress (40 mM NaCl), Trichoderma increased pak choi leaf chlorophyll content. Under moderate-salinity stress (80 mM NaCl) condition, the addition of Glomus sp. increased lettuce fresh weight and leaf area, and B/P/T mix increased pak choi leaf area. In conclusion, using the selected PGPMs in low to moderate-salinity stress could increase lettuce and pak choi growth and quality parameters. These results have some practical applications in the future when more saline water is used for production.

Liquid-nitrogen-free CTAB DNA extraction method from silica-dried specimens for next-generation sequencing and assembly

Next-generation sequencing requires intact and high-quality DNA. However, typical liquid-nitrogen DNA extraction methods are expensive and not practical for field sample collections. Hence, we present a cost-effective method for DNA extraction from silica-dried leaf samples, eliminating the need for liquid nitrogen. Two protocols were evaluated to determine the effectiveness of grinding dried plant samples without liquid nitrogen in comparison to the standard protocol for tissue homogenization and cell lysis. Protocol 1 involved grinding fresh leaf samples with liquid nitrogen, while Protocol 2 entailed incubating dried plant samples at-20 °C for 1 h before grinding in the absence of liquid nitrogen. Both protocols produced comparable DNA yields with an average A260/A280 ratio of 1.78±0.02, suitable for short- and long-read sequencing. Using Protocol 2, we successfully assembled ten plastomes. It also demonstrated versatility as comparable DNA quality was obtained from dried mollusks and actinomycetes, resulting in thesuccessful assembly of two complete mitochondrial genomes. The protocol is advantageous for research workflows involving the collection of samples in the field as a long-term source of genetic material.•Drying: Fresh samples were silica-dried at silica-to-sample ratio of 2:1.•Pre-lysis: Dried samples were frozen at −20 °C for 1 hour before grinding.•Frozen samples were subjected to tissue homogenization followed by the standard CTAB DNA extraction.

Optimizing phosphorus fertilization for two commonly grown ground covers: creeping jenny and periwinkle

Growing nursery plants in containers usually requires the addition of fertilizer to achieve optimal growth. However, misconceptions among growers often lead them to apply quantities of fertilizers exceeding the recommended dose, resulting in nutrient runoff and increased production costs. Reducing fertilizer application is beneficial to have healthy plants and environment as well as to save on fertilizer costs. In this study, we wanted to determine the minimal level of phosphorus (P) required to grow bedding plants i.e. Lysimachia nummularia ‘creeping jenny’ (moneywort) and Vinca minor ‘Bowles’ (periwinkle) in nursery. The plants were grown within a greenhouse in a mixture of peat moss, vermiculite, and rice hulls (3:1:1 v/v) substrates and subjected to various P treatments. The treatments included single dose of 1, 3, or 6 mg/L of P, double doses of 3 or 6 mg/L of P, or a control that received 6 mg/L of P at each irrigation. The results indicated that single dose of 6 mg/L of P had similar fresh weight compared to control in both taxa. Reducing P application further to a single dose of 1 and 3 mg/L reduced fresh weight and leaf area in both groundcovers however did not reduce their stem length or physiological parameters such as Soil Plant Analysis Development (SPAD) and normalized difference vegetation index (NDVI). Phosphorus concentration in the plant tissue at maturity was lower in all treatments than control for both the taxa. During the study period, control plants received 26 times the dose of P compared to plants receiving 6 mg/L of P once. Therefore, this study outlines the possibility of reducing P fertilizer application while growing these groundcovers in nurseries for sale.

MORPHO-ANATOMICAL PROFILE OF ELAEOCARPUS TUBERCULATUS ROXB

Objective: The anatomical studies of Elaeocarpus tuberculatus Roxb. (Elaeocarpaceae), an indigenous medicinal plant in the Indian pharmacopoeia. Methods: Fresh leaf and stem samples of E. tuberculatus were scrutinized macroscopic and microscopically by standard procedures. Results: Macroscopically, the leaves are simple, long, coriaceous, and rusty below, base obtuse, margin serrate, apex obtuse and petiole 3 cm long. Microscopically, the leaf showed the presence of four prominent collateral bundles. Paracytic stomata were distributed in the abaxial epidermis whereas the adaxial epidermis was apostomatic. The petiole was four-angled including wing bundles. The stem exhibited fissured periderm, wide secondary phloem, secondary xylem with incipient growth ring and abundant calcium oxalate crystals. Powdered microscopic study of the leaf revealed the presence of long whip-like epidermal trichomes, crystals, vessel elements showing pits and fiber-like sclereids. The stem powder showed thick-walled fibers with wide lumen, narrow fibers and a sclerenchyma cell showing wide pits. Conclusion: The results of the study can serve as a valuable source of information and provide suitable standards for the identification of E. tuberculatus in future investigations and applications