The accurate identification of walnut (Juglans regia L.) varieties is crucial for nurseries to detect any mistakes that might occur during vegetative propagation, enabling them to certify their plant material. It is also essential for farmers to resolve any uncertainties about the plant material used in orchard plantations. The availability of a practical, accurate, and reliable tool for walnut genotyping will be of obvious interest. Traditional genotyping methods, such as microsatellite (Simple Sequence Repeats – SSR) analysis, are often too time-consuming and expensive for routine use. This study introduces High-Resolution Melting (HRM) analysis as an innovative, cost-effective, and rapid technique for effective walnut variety genotyping.

HRM analysis utilizes real-time PCR to amplify variable genomic regions, followed by melting curve analysis to distinguish between different varieties. This method streamlines the genotyping process, making it both accessible and practical for routine applications. PCR conditions were optimized to improve the accuracy and efficiency of HRM analysis, and its effectiveness was demonstrated by comparing the results with those obtained through SSR analysis.

Leaves collected from four J. regia varieties (Tulare, Lara, Howard, and Chandler), grown in different orchards in the Alentejo region of Portugal, were used as plant material to establish the methodology. Polymorphic regions corresponding to thirteen SSR loci were examined, and three proved to be effective in differentiating the varieties (WGA202, WGA321, and WGA376).

Validation via conventional microsatellite analysis substantiated the reliability of the HRM-based method. The implementation of HRM analysis can mitigate the risk of varietal mixing, thereby ensuring superior quality control and compliance with market standards. Furthermore, future studies could extend this technique to the identification of walnut varieties at the fruit level, facilitating the correct identification of market-bound fruits and ensuring higher quality for consumers.

Introduction: The third step in histidine degradation is catalysed by imidazolonepropionase. It catalyses the conversion of 4-imidazolone-5-propionic acid to produce N-formimino-l-glutamic acid by hydrolyzing carbon—nitrogen bonds. The histidine is a very expensive amino acid inside the cell and its degradation is a very conserved process. To date, very few reports exist regarding the structure of bacterial imidazolonepropionase, but no reports have been published regarding the comparative structure and sequence analysis of this enzyme from bacterial sources.

Methods: An in silico study has been conducted to characterize imidazolonepropionase from gram-positive Bacillus subtilis and gram-negative Agrobacterium fabrum.

Results: The sequence analysis revealed that more charged residues are present in Bacillus subtilis. These charged residues help in the increment of the polarity and hydrophilicity of Bacillus subtilis. The formation of intra-protein interactions was also high in gram-positive species. Interestingly, both species have an almost equal abundance of aromatic amino acids in their sequences, but the formation of aromatic—aromatic interactions was high in Bacillus subtilis. Finally, the molecular dynamics simulation study revealed that imidazolonepropionase from Bacillus subtilis was more stable and compact than Agrobacterium fabrum.

Conclusion: The imidazolonepropionase from Bacillus subtilis was more stable than Agrobacterium fabrum. Due to the presence of higher stable imidazolonepropionase in Bacillus subtilis, it can use histidine more efficiently.

In vitro propagation offers disease-free material in ginger (Zingiber officinale Rosc.), a plant of significant medicinal and commercial value. This study aims to investigate the anatomical and biochemical characteristics of in vitro and in vivo plants of ginger. Fifty-day-old in vitro (developed from tissue culture lab maintained at 25 ± 2°C and 90-92% RH over 14 h photoperiod at 3000 lx) and in vivo plants (pro-tray plants grown in polyhouse maintained at 25 ± 2°C and 60-70% RH) of the ginger variety IISR Varada were taken to be studied at the ICAR—Indian Institute of Spices Research, Kerala, India. Transverse sections of leaves, the pseudo-stem, and the rhizomes of the in vitro and in vivo ginger plants were taken and visualized under light microscopy.
In vitro and in vivo ginger possess similar leaf structures but differ in their spongy parenchyma thickness, the number of stomata, oil cells, and air canals, and their vascular bundle distribution. The in vitro pseudo-stems had a closely bound leaf sheath and epidermis unlike their in vivo counterparts. Rhizome analysis revealed larger vascular bundles in the in vivo ginger (normal rhizomes produced at farm) and higher starch and sugar content in the in vitro rhizomes (micro-rhizomes developed at tissue culture lab).
Biochemical analysis revealed that the total chlorophyll and carotenoid content was significantly higher in the in vivo ginger (1.1989 mg/g and 67.24 mg/g) compared to the in vitro ginger (0.7173 mg/g and 50.87 mg/g, respectively). Furthermore, the enzyme activities of peroxidase (0.063 mg/g), catalase (108.248 U/g), and superoxide dismutase (0.367 U/g); the starch content (22.84%); and the total soluble sugars (0.093%) were higher in the in vitro ginger plants.
Hence, the anatomical and biochemical variations observed in the different parts of the in vitro and in vivo ginger plants may be due to the differences in the growing conditions and the media used in the in vitro conditions.

Seed quality affects crop establishment and productivity. In addition, the use of good-quality seed is an essential prerequisite for sustainable crop production including groundnuts. Assessing germinability and electrical conductivity provides early evidence of the production potential of a given crop variety or genotype. Therefore, this study assessed the germinability and electrical conductivity of seeds of three groundnut varieties. A laboratory experiment arranged in a Completely Randomized Design (CRD), replicated three times, was conducted at the Faculty of Agriculture, Kyambogo University, in 2020. Seeds of groundnut varieties Igola, Serenut 1, and Serenut 2 were tested, and data were collected on germination percentage and electrical conductivity. Analysis of variance (ANOVA) was performed using Genstat and means were separated using the least significance test at a 5% probability level. Germination percentage and electrical conductivity significantly (p<0.05) differed among the groundnut varies, with Igola recording the highest germination percentage, followed by Serenut 1, and the lowest was in Serenut 2. The highest electrical conductivity was recorded in Serenut 1 and the lowest in Igola. Since Igola had one of the lowest electrical conductivities and the highest germination percentage, it was concluded that Igola was the most promising variety from the studied ones. This study recommends Igola to be evaluated further for growth and yield performance.

Halophytes have evolved a variety of strategies to tolerate elevated salinity levels. Cakile maritima, a succulent annual halophyte from the Brassicaceae family, is widely distributed across global coastal environments, primarily thriving in foreshores. This species exhibits optimal growth under moderate saline conditions and is capable of completing its life cycle in environments with salt concentrations reaching up to 500 mM NaCl. The response of Cakile maritima to salinity during its vegetative development has been extensively documented, with various mechanisms identified, including anatomical and morphological adaptations, ion transport regulation, osmolyte biosynthesis, and activation of antioxidative mechanisms. However, there is limited knowledge regarding the effects of salt stress on its reproductive development. Preliminary results indicated a pronounced negative impact of salinity on flowering and fruit production in Cakile maritima. To elucidate the detrimental effects of salinity on the plants' reproductive capacity, we conducted a histological analysis of anther anatomy at various developmental stages, along with assessments of pollen viability and germination potential. These analyses were performed on plants irrigated with either freshwater or saline water with increasing concentrations of NaCl. The gametophyte development in halophytes, and particularly in Cakile maritima, remains an underexplored research area. This study aims to provide new insights into the reproductive adaptations of halophytes by examining the impact of salinity on the biology of male gametophytes, thus contributing to our understanding of their survival in extreme environments.

The coevolution process has shaped the diversity and complexity of life on Earth. Coevolution involves the interaction between two entities in a highly specific process because selective pressures in one of the entities drive the evolution of the other. Moreover, coevolution is reciprocal and simultaneous, as evolution occurs in both entities at the same time. At the molecular level, molecular coevolution occurs in systems where molecules interact closely (e.g., enzymes and their substrates, proteins and their binding partners, transcription factors and their specific binding motifs on DNA, etc.). Many factors play a pivotal role in shaping molecular coevolution, such as drift intensities, selection, and mutation rates. Intramolecular coevolution occurs between sites within a single molecule, while intermolecular coevolution takes place between sites of two interacting molecules. Identifying coevolving sites is crucial for understanding protein-protein interactions, drug resistance, the evolutionary arms races between hosts and pathogens, and for enhancing molecular activity.

Plant response mechanisms to salinity stress are diverse and complex. Among them, those involved in ion homeostasis and compartmentalisation play a key role in isolating and eliminating toxic ions. The main strategies are the expression of Salt Overly Sensitive (SOS) genes, the High-Affinity Potassium Transporters (HKTs), the Na⁺/H⁺ Antiporter (NHX), and the proton pumps. The aim of this work is, first, to identify coevolving amino acids within these proteins and, second, to find if there exist any coevolving sites between them. We used CAPS, a software to identify co-evolution between amino acid sites by measuring the correlated evolutionary variation at those sites, which further removes the phylogenetic and stochastic dependencies between sites. The software uses a reference sequence for which the 3D protein structure is available. NHX1, SOS1, SOS2, and HKT1 were analysed, and we found pairs of sites under strong intramolecular co-evolution as well as intermolecular interactions.

Salinity is considered the most limiting abiotic stress affecting both ecosystems and global agriculture by hampering plant growth and development, thus seriously limiting plant productivity and survival. Therefore, investigating the response to salt stress in plants is pivotal to understanding how ecosystems will face the effects of climate change. Salt stress is a very likely event in coastal ecosystems. The Mediterranean coastal ecosystems, such as salt marshes, host valuable plant species with heterogeneous degrees of tolerance to salt. La Albufera Natural Park in Valencia, Spain, is home to several species of Plantago known for their ability to withstand salt stress, such as P. coronopus and P. crassifolia, which are more tolerant than species like P. lanceolata or P. major. Moreover, some Plantago species are known to host diverse microbial endophyte species with potential beneficial effects.

The main objective of this research is to understand the molecular dynamics of salt stress gene expression when exposing Plantago species with different ecological strategies (glycophyte or halophyte) to 400 mM NaCl salt stress. Additionally, we will investigate the potential role of local soil microbiota on the plants’ response. RNA expression of salt-tolerant genes will be quantified at times 0, 2, 4, 8, 24 h, and when plants start to wilt. Morphological, physiological, and molecular stress markers will be recorded for each plant.

We found a heterogenous response of the biochemical markers of stress depending on the treatment or the Plantago species. Moreover, we found several fungal endophytes associated with the roots and leaves from wild P. crassifolia. We further discuss the different levels of gene expression.

Oxidative stress plays a pivotal role in the onset and progression of chronic and metabolic diseases, including diabetes, obesity, and cancer, due to the excessive production of reactive oxygen species, which cause damage to essential biomolecules and impair critical cellular functions [1]. Antioxidants such as polyphenols can mitigate or delay this damage, driving growing interest in the use of polyphenol-rich extracts as natural functional food ingredients. Notably, polyphenols can be obtained from the massive amounts of undervalued plant biomass generated after crop harvesting [2]. Therefore, this study was caried out to valorize sweet pepper (Capsicum annuum L.) agricultural by-products as a source of bioactive polyphenols, in line with United Nations' 2030 Sustainable Development Goals. Hydroethanolic extracts were prepared from these plant by-products [2], and their phenolic profile was characterized using HPLC-DAD-ESI/MS. Additionally, the extracts' antioxidant, antidiabetic, anti-obesity, anti-inflammatory, and cytotoxic properties were assessed through in vitro cell-based and enzyme inhibition assays. Chromatographic analysis revealed a phenolic profile consisting mainly of phenolic acids, such as chlorogenic acid, and O-glycosylated flavones, specifically luteolin and apigenin. The extracts showed notable antioxidant and anti-diabetic properties, along with cytotoxicity against certain tumor cell lines. In conclusion, sweet pepper crop by-products are a promising source of bioactive compounds that can provide functional properties to foods and related products. Furthermore, this approach to resource circularity is crucial for promoting sustainable production and consumption patterns, contributing to a more eco-friendly and efficient use of agricultural resources. However, ensuring that the final products are free from pesticide residues and other contaminants will be essential for guaranteeing food safety.

Introduction: The fragmentary and whole substitution of wheat flour with flour blends is an alternative approach for producing cheaper, nutrient-rich, and comparatively advantageous gluten-free foods through fermentation. Methods: Dry samples of sweet potato, pigeon pea, and maize botanicals were purchased from local vendors, and authenticated and processed before spontaneous fermentation at room temperature. The pH and microbiological patterns of the fermenting botanicals were evaluated every 12 hours for 72 hours, using standard test protocols. Results: It revealed that the rates of growth of isolated microorganisms were affected by pH; all the botanicals fermented had a reduction in their pH values. Acids were produced during fermentation, leading to a reduction in pH. Bacteria growth on the fermenting samples on nutrient agar reveals that the bacterial load increased with fermentation time, from 7.52 Log10 CFU/g to 10.6 Log10 CFU/g (sweet potato); 6.3 Log10 CFU/g to 10.54 Log10 CFU/g (pigeon pea), and 6.3 Log10 CFU/g to 10.54 Log10 CFU/g (maize). On MacConkey agar, the bacterial load on all samples started after 24 hours of fermentation, peaked at 48 hours, and reduced gradually towards 72 hours of fermentation. There was increase in fungal growth with time from 0 to 36 hours across all samples. The microorganisms isolated can be categorized into lactic acid bacteria, spore formers, Enterobacteriaceae, Staphylococcace, yeast, and moulds. Conclusion: Fermentation of botanicals over 72 hours results in organic acid formation, which lowers pH; this attribute helps in checkmating undesirable microorganisms capable of affecting the production of gluten-free flours with good keeping qualities.

Plants of Ananas comosus (cultivar Smooth Cayenne) were grown in pots filled with a commercial substrate and exposed to different water content treatments: 100% field capacity (FC), 50% FC, and 25% FC. The experiment was conducted under controlled greenhouse conditions for three months. Every month, relative water content (RWC), NDVI, proteolytic activity (PA), and protein content (PC) were measured in the "D" leaves. Additionally, proteolytic activity and protein content were measured monthly in the stem and leaves. RWC showed no significant differences during the first month. However, by the second month, the 50% and 100% FC treatments had significantly higher values than the 25% FC treatment. By the third month, all three treatments were statistically different, with the 100% FC treatment having the highest RWC. In the first month, the 100% treatment showed statically higher NDVI values than the 50% and 25% FC treatments. By the second and third months, the three treatments were statistically different, with average NDVI values of 0.57, 0.67, and 0.72 for the 25%, 50%, and 100% FC treatments, respectively. PA and PC increased in both the leaves and stems as drought stress intensified, with a significant increase from 100% to 25% FC over the course of the experiment. The non-destructive indicator (NDVI) showed differences in the first month, while the destructive indicators (RWC, PA, and PC) in both organs exhibited statistical differences by the second month of the experiment. The NDVI proved to be a highly sensitive indicator of drought stress in pineapple plants and did not require the destruction of the plant.