Biotechnological Applications Research Articles

Characterization of a novel salt- and solvent-tolerant esterase Dhs82 from soil metagenome capable of hydrolyzing estrogenic phthalate esters

Esterases that can function under extreme conditions are important for industrial processing and environmental remediation. Here, we report the identification of a salt- and solvent-tolerant esterase, Dhs82, from a soil metagenomic library. Dhs82 prefers short-chain p-nitrophenyl (p-NP) esters and exhibits enzymatic activity up to 1460 ± 61 U/mg towards p-NP butyrate. Meanwhile, Dhs82 can catalyze the hydrolysis of dialkyl phthalate esters, especially the widely-used diethyl phthalate (DEP), dipropyl phthalate (DPP) and di-n-butyl phthalate (DBP). Importantly, as an acidic protein with negative charges dominating its surface, Dhs82 is highly active and extraordinarily stable at high salinity. This property is quite rare among previously reported esterases/hydrolases capable of degrading phthalate esters (PAEs). In addition, Dhs82 activity can be significantly enhanced in the presence of solvents over a concentration range of 10–30 % (v/v). Notably, Dhs82 also showed high stability towards these solvents and solvent concentrations as high as 50–60 % (v/v) are required to inactivate Dhs82. Furthermore, molecular docking revealed the key residues, including the catalytic triad (Ser156, His281, and Asp251) and the surrounding Gly84 and Gly85, involved in the interaction of Dhs82 with DBP, depicting how Dhs82 degrades PAEs as a family IV esterase. Together, these diverse properties make Dhs82 a valuable candidate for both basic research and biotechnological applications.

WITHDRAWN: The enhanced heat transfer and flow dynamics of tangent hyperbolic and Newtonian nanofluids under inclined variable magnetic field

This paper aims to analyze the thermal conductivity of tangent hyperbolic non-Newtonian fluids and Newtonian nanofluids over a porous, inclined, stretching sheet with motile microorganisms. It also examines the impact of inclined variable magnetic field, activation energy, thermal radiation, thermophoresis and Brownian motion. The novelty of this research lies in the simultaneous use of inclined geometry and an inclined variable magnetic field. The findings optimize environmental and food processing applications, drug delivery, thermal management, lubrication, and enhanced oil recovery. A far-reaching analysis compares the behaviors of tangent hyperbolic fluid with Newtonian fluid. Appropriate similarity functions are used to convert the governing system of nonlinear PDEs for tangent hyperbolic fluid to dimensionless ODE. MATLAB’s boundary value solver BVP4C is then used to solve these ODEs numerically. This study explores various characteristics, including temperature, velocity, concentration, and microorganism distribution. In the essence of this computational study, the impacts of inclined angle, porosity, mixed convection, Hartmann number and Weissenberg number on the velocity of the fluid are analyzed. The main findings are that higher Prandtl numbers reduce heat transfer rates, while thermophoresis and Brownian motion enhance heat flow. Lewis and Peclet numbers show declining tendencies in motile microorganisms, with magnetic fields influencing their distribution within the fluid; this is useful in biotechnology and environmental applications. Consequently, the effect is greater in the case of non-Newtonian fluids which depend more on the inclination angle, inclined variable magnetic field strength, thermal conductivity and activation energy when compared with the Newtonian fluids.

A putative hemicellulose gene promoter drives high levels of expression in guard cells and stem epidermis of poplar and highlights hormone regulation of cell wall formation

Forest trees generate substantial quantities of organic carbon, recognized as a renewable and economically viable resource for applications in bioenergy and industry. This study specifically focuses on xylan, a major hemicellulose component in poplar, and on the role of PtrPARVUS2—a member of the glycosyltransferase 8 family genes involved in xylan biosynthesis. PtrPARVUS2 is hypothesized to exhibit tissue-specific activity, and here we assessed its promoter via bioinformatic analysis, revealing motifs associated with responsiveness to hormones including Gibberellic Acid (GA), Methyl Jasmonate (MeJA), Auxin, and Abscisic Acid (ABA), among others. Treatment with hormones, including ABA, 1-naphthaleneacetic acid, a synthetic auxin, MeJA, and GA, demonstrated a swift inhibition of PtrPARVUS2 transcription. Analysis of expression patterns in wildtype trees indicated heightened expression in vascular tissues, particularly the bark. Transgenic lines expressing Enhanced Green Fluorescent Protein and β-glucuronidase under the control of the PtrPARVUS2 promoter confirmed tissue-specific activity, particularly in guard cells, stem epidermis, and developing xylem. These results offer understanding into the potential functions of PtrPARVUS2 in terms of phytohormone response and the formation of guard cell walls. The investigation introduces a valuable promoter for specific biotechnological applications in guard cells and stem epidermal cells. Additional research is required to elucidate the precise regulatory mechanisms of PtrPARVUS2 in the future.

Fishpond microbiota, their synergism with fish and potential biotechnological applications

Fishpond microbiota, their synergism with fish and potential biotechnological applications

Development and characterization of microencapsulated Pichia kluyveri CCMA 0615 with probiotic properties and its application in fermented beverages

The study aimed to develop innovative microencapsulated formulations of strains with probiotic attributes, Pichia kluyveri CCMA 0615 and Saccharomyces cerevisiae CCMA 0732. The yeasts (8 log CFU/mL) were microencapsulated by spray drying technique using whey powder (WP - 15 %, 20 %, and 30 %) and sodium alginate (ALG - 1 %). The microcapsules and cell viability were characterized during two months of storage (4 °C and 25 °C). The selected formulations were applied to functional beverage fermentation, and viability and survival in the simulated gastrointestinal tract (GIT) were performed. The viability of yeasts microencapsulated by the spray drying method was shown to be dependent on the strain and encapsulating matrix used, ranging from 84 to 99 %. P. kluyveri required refrigeration when storing microcapsules. In functional beverage fermentation, microencapsulated yeast maintained the same fermentative profile with carbohydrate consumption, production of lactic acid (0.30 to 1.10 g/L) and alcohol (0.2 to 1.61 g/L), and greater viability during storage. Finally, the microencapsulation of P. kluyveri with 15 % WP + 1 % ALG maintained high viability under GIT conditions, whether exposed independently (>84 %) or incorporated into a food matrix (>94 %). The study demonstrated that this innovative microencapsulation of probiotic yeasts increases their viability, improves biotechnological application, and facilitates efficient delivery of probiotics to the host.

Optimisation of coumaric acid production from aromatic amino acids in Kluyveromyces marxianus

Yeasts are attractive hosts for the production of heterologous products due to their genetic tractability and relative ease of growth. While the baker’s yeast Saccharomyces cerevisiae is a powerful workhorse of the biotechnology industry, the species has metabolic limitations and it is critical that we develop alternative platforms that will facilitate the development of bioprocesses that rely on sustainable feedstocks. In this study, we used synthetic biology tools to construct coumaric acid–producing strains of Kluyveromyces marxianus, a yeast whose physiological traits render it attractive for biotechnology applications. Coumaric acid is a building block in the synthesis of many different families of aromatics and is a key precursor for the synthesis of complect phenylpropanoid molecules, including many flavours and aromas. The starting point for this work was a K. marxianus chassis strain that has increased flux towards the synthesis of tyrosine and phenylalanine, the aromatic amino acids that can serve as starting points for coumaric acid synthesis. Following principles of synthetic biology, a modular approach was taken to identify the best solution to different metabolic possibilities and these were then combined in different ways. For the first step, it was established that the route from phenylalanine was superior to that from tyrosine and the combined overexpression of PlPAL, AtC4H and AtCPR1 delivered the highest yield of coumaric acid. Next, it was established that while Pdc5 and Aro10 both had phenylpyruvate decarboxylase activity, inactivation of ARO10 was sufficient to prevent flux loss in the pathway. Since phenylalanine is the starting point, efforts were made to improve efficiency of its production. It was found that glutamate was a preferred nitrogen source for coumaric acid production, and this knowledge was used to engineer a strain that overexpressed S. cerevisiae GDH1 and delivered higher yields of coumaric acid. Ultimately, this strategy led to the development of strains that has yields of up to 48mg coumaric acid /g glucose. Strains were evaluated in bioreactors to investigate the effects of different process parameters. These analyses indicated that engineered strains face some redox balance challenges and further work will be required overcome these to develop strains that can perform well under industrial conditions.

The fnr-like mutants confer isoxaben tolerance by initiating mitochondrial retrograde signalling.

Isoxaben is a pre-emergent herbicide used to control broadleaf weeds. While the phytotoxic mechanism is not completely understood, isoxaben interferes with cellulose synthesis. Certain mutations in cellulose synthase complex proteins can confer isoxaben tolerance; however, these mutations can cause compromised cellulose synthesis and perturbed plant growth, rendering them unsuitable as herbicide tolerance traits. We conducted a genetic screen to identify new genes associated with isoxaben tolerance by screening a selection of Arabidopsis thaliana T-DNA mutants. We found that mutations in a FERREDOXIN-NADP(+) OXIDOREDUCTASE-LIKE (FNRL) gene enhanced tolerance to isoxaben, exhibited as a reduction in primary root stunting, reactive oxygen species accumulation and ectopic lignification. The fnrl mutant did not exhibit a reduction in cellulose levels following exposure to isoxaben, indicating that FNRL operates upstream of isoxaben-induced cellulose inhibition. In line with these results, transcriptomic analysis revealed a highly reduced response to isoxaben treatment in fnrl mutant roots. The fnrl mutants displayed constitutively induced mitochondrial retrograde signalling, and the observed isoxaben tolerance is partially dependent on the transcription factor ANAC017, a key regulator of mitochondrial retrograde signalling. Moreover, FNRL is highly conserved across all plant lineages, implying conservation of its function. Notably, fnrl mutants did not show a growth penalty in shoots, making FNRL a promising target for biotechnological applications in breeding isoxaben tolerance in crops.

Characterization of hemicellulose in sacred lotus (Nelumbo nucifera Gaetn.) petiole during xylogenesis

Hemicelluloses play a crucial role in connecting cellulose and lignin within the plant cell wall and find extensive biotechnological applications. There is a notable gap in research concerning the hemicellulose from Nelumbo nucifera, the basal eudicot adapted to aquatic environments. To fill this gap, hemicellulose characteristics from the apical to the basal segments of petioles from 4-month-old N. nucifera plants were examined. Results showed that during the initial phase of xylogenesis, xyloglucan predominated in the hemicellulose fraction, while later stages were gradually dominated by 4-O-methyl-D-glucurono-D-xylan (MGX). Having a typical tetrasaccharide reducing end, the glucuronoxylan in N. nucifera was also shown with a typical the MeGlcA on the C2 Xyl carbon sidechain. The total degree of acetylation increased from apex (0.45) to base (0.55) of petiole. Notably, no arabinosyl side-chains were detected in the N. nucifera xylan, suggesting that N. nucifera hemicellulose aligns most closely with dicot wood rather than the non-commelinid monocot (grass). Transcriptomic analysis also indicated that the middle and basal region exhibited higher xylan synthesis activity. This study contributes new evidence supporting the conservation of dicotyledonous hemicellulose during evolution.

Hydrogen production capabilities of lichens micro-ecosystem under extreme salinity, crystalline salt exposure, and simulated Mars-like conditions

This work aims to demonstrate the extremophilic behavior of the lichen Pleurosticta acetabulum at extreme salinities, while maintaining its metabolic capacity to produce hydrogen. Lichen is a special micro-ecosystem that includes mostly a fungus and a green alga or cyanobacterium, as well as a microbiome. The peculiarity of this symbiotic system is its ability to dry out completely and stay inactive to survive harsh conditions. Lichens that had been dehydrated for six months revived quickly when rehydrated, restoring their photosynthetic efficiency and ability to produce hydrogen. The lichen microbiome was crucial for hydrogen production, especially through dark fermentation. The experiments of this work showed that lichen during its exposure to different salinity conditions (0%NaCl – control, 3,5%NaCl – sea salt concentration, 36%NaCl – saturated salt concentration), but also after exposure to crystalline salt (100%NaCl) could maintain the structure and the functionality of its photosynthetic apparatus. This was tested using chlorophyll a fluorescence induction measurements. Based on the results of thermal conductivity gas chromatography (GC-TCD) for the determination of hydrogen production, it was shown that despite being exposed to extreme salinity conditions, lichens maintained its ability to produce hydrogen. The experimental combination of lichen exposure to extreme salinities (up to 100% NaCl), with an extreme atmosphere (100% CO2) and low atmospheric pressure (<10mbar), simulating Mars conditions, highlighted the functional potential of the lichen for survival in a Mars-like environment. This lichen’s ability to withstand extreme conditions and to produce large amounts of hydrogen, makes it a promising candidate for future biotechnological applications, even in challenging environments like Mars, opening new astrobiological and astrobiotechnological perspectives.

CRISPR-Cas9: Revolutionizing Gene Therapy for Genetic Disorders

Faulty or mutated genes cause multiple human disorders such as cancer, neurogenerative disorders, and cardiovascular diseases. These mutations in genes are usually inherited. Many treatments have been established to cure such diseases, but gene therapy is the most promising strategy. It’s an application of biotechnology that is based on correcting and replacing the mutated gene with the healthy gene, providing the genes for the proper expression of desired proteins required for curing the disease. In clinical settings, gene therapy has proved itself as a very promising treatment but has some drawbacks. Traditionally, these methods involved viral vectors that are used to deliver correct genes and replace them with the mutated genes, in patients. These approaches have somehow potential to cure diseases but often off-target effects, limitations in the editing process, and immune responses caused by patients’ immune systems are the main challenges that are the main shortcomings of this method. However, the discovery of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated nuclease protein 9 (Cas9) genome editing system (CRISPR/Cas9) in 2012 and its development have increased the value of gene therapy in the therapeutic world [1]. CRISPR/Cas9 is a revolutionizing tool that has been in gene therapy for knocking in and out gene to correct the mutation associated with many genetic diseases. This system has evolved greatly and has many isomers using different strategies to improve both applied, basic research and its clinical application. The ability of CRISPR-Cas9 to target various genetic diseases is one of its greatest advantages. Mutation in single gene results in monogenic diseases such as cystic fibrosis, muscular dystrophy, and sickle cell anemia, and this system has shown the potential to edit or fix the defective genes, causing these diseases. Even in the case of complex disorders caused by the mutation in multiple genes such as several types of cancer, and cardiovascular diseases, CRISPR has shown very positive results. The application of CRISPR-Cas9 in gene therapy is very promising and has tremendous potential to give cures for many complex disorders. The ethical issue regarding the editing of human DNA and the inheritance of modified DNA into the next generation is still under discussion and causing hurdles in uncovering the full potential of this system. Researchers are working to increase its efficiency and specificity to reduce the off-target sequences so that only targeted modification can be achieved. Moreover, they are trying to give possible results to reduce ethical concerns. CRISPR has evolved greatly and has many isomers using different strategies to improve both applied, basic research and its clinical application in the future.

Recent Advances in Genetic Engineering Strategies of Sinorhizobium meliloti.

Sinorhizobium meliloti is a free-living soil Gram-negative bacterium that participates in nitrogen-fixation symbiosis with several legumes. S. meliloti has the potential to be utilized for the production of high-value nutritional compounds, such as vitamin B12. Advances in gene editing tools play a vital role in the development of S. meliloti strains with enhanced characteristics for biotechnological applications. Several novel genetic engineering strategies have emerged in recent years to investigate genetic modifications in S. meliloti. This review provides a comprehensive overview of the mechanism and application of the extensively used Tn5-mediated genetic engineering strategies. Strategies based on homologous recombination and site-specific recombination were also discussed. Subsequently, the development and application of the genetic engineering strategies utilizing various CRISPR/Cas systems in S. meliloti are summarized. This review may stimulate research interest among scientists, foster studies in the application areas of S. meliloti, and serve as a reference for the utilization of genome editing tools for other Rhizobium species.

Design of a Genetically Programmable and Customizable Protein Scaffolding System for the Hierarchical Assembly of Robust, Functional Macroscale Materials.

Inspired by the properties of natural protein-based biomaterials, protein nanomaterials are increasingly designed with natural or engineered peptides or with protein building blocks. Few examples describe the design of functional protein-based materials for biotechnological applications that can be readily manufactured, are amenable to functionalization, and exhibit robust assembly properties for macroscale material formation. Here, we designed a protein-scaffolding system that self-assembles into robust, macroscale materials suitable for in vitro cell-free applications. By controlling the coexpression in Escherichia coli of self-assembling scaffold building blocks with and without modifications for covalent attachment of cross-linking cargo proteins, hybrid scaffolds with spatially organized conjugation sites are overproduced that can be readily isolated. Cargo proteins, including enzymes, are rapidly cross-linked onto scaffolds for the formation of functional materials. We show that these materials can be used for the in vitro operation of a coimmobilized two-enzyme reaction and that the protein material can be recovered and reused. We believe that this work will provide a versatile platform for the design and scalable production of functional materials with customizable properties and the robustness required for biotechnological applications.

Encoding extracellular modification of artificial cell membranes using engineered self-translocating proteins

The development of artificial cells has led to fundamental insights into the functional processes of living cells while simultaneously paving the way for transformative applications in biotechnology and medicine. A common method of generating artificial cells is to encapsulate protein expression systems within lipid vesicles. However, to communicate with the external environment, protein translocation across lipid membranes must take place. In living cells, protein transport across membranes is achieved with the aid of complex translocase systems which are difficult to reconstitute into artificial cells. Thus, there is need for simple mechanisms by which proteins can be encoded and expressed inside synthetic compartments yet still be externally displayed. Here we present a genetically encodable membrane functionalization system based on mutants of pore-forming proteins. We modify the membrane translocating loop of α-hemolysin to translocate functional peptides up to 52 amino acids across lipid membranes. Full membrane translocation occurs in the absence of any translocase machinery and the translocated peptides are recognized by specific peptide-binding ligands on the opposing membrane side. Engineered hemolysins can be used for genetically programming artificial cells to display interacting peptide pairs, enabling their assembly into artificial tissue-like structures.

A DNA Aptamer for 2-Aminopurine: Binding-Induced Fluorescence Quenching.

2-Aminopurine (2AP) is a fluorescent analog of adenine, and its unique properties make it valuable in various biochemical and biotechnological applications. Its fluorescence property probes local dynamics in DNA and RNA because stacking with the surrounding bases quench its fluorescence. 2AP-labeled DNA or RNA sequences have been used for the detection of genetic mutations, viral RNA, or other nucleic acid-based markers associated with diseases like cancer and infectious diseases. In this study, we isolated aptamers for 2AP using the library immobilization capture-SELEX technique. A dominating aptamer family was isolated after 15 rounds of selection. The Kd values for the most abundant 2AP1 aptamer are 209 nM in a fluorescence assay and 72 nM in an isothermal titration calorimetry test. A 32 nM 2AP limit of detection was tested based on its intrinsic fluorescence change upon aptamer binding. Additionally, we conducted some mutation analysis. Furthermore, we tested the selectivity of this aptamer and discovered that it can bind adenine and adenosine with approximately 100-fold lower affinity than 2AP.

Studying the Antimicrobial Effects of Silver Nanoparticles Coated with Alginate Biosynthesized by Dulcicalothrix alborzica

Introduction: Nowadays, Cyanobacteria are one of the important candidates in the green biosynthesis of nanoparticles. Considering the detrimental effects of chemically synthesized nanoparticles, the aim of this study was the biosynthesis and antimicrobial activity of nanoparticles by aquatic cyanobacterium. Methods: Silver nanoparticles (AgNPs) were biosynthesized using three different approaches: wet biomass, boiling, and extracellular polysaccharides (EPS). Alginate coating was employed to enhance the nanoparticles' stability. Characterization of the nanoparticles was performed using UV-vis spectroscopy, Fourier transform infrared (FTIR) analysis, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and zeta potential measurements. The antimicrobial properties of the nanoparticles were evaluated against fish pathogenic bacteria and fungi. Statistical analyzes were performed with SPSS version 16 software, and the significant difference between the means was performed with one-way analysis of variance with 95% confidence limits and Tukey's test, and the results were drawn as a graph with Excel software. Results: The UV-vis spectroscopy results confirmed the synthesis of AgNPs in all three methods. FTIR analysis revealed similar spectra for all three methods, indicating comparable purity and production of similar compounds. Electron microscope images showed the sphericity of nanoparticles prepared by boiling method, and the average diameter of uncoated and alginate coated nanoparticles was 38 and 180.04 nm, respectively. Zeta potential analysis indicated a positive surface charge on the nanoparticles. The highest zone of inhibition was observed for AgNPs synthesized by the boiling method and coated with alginate. Similar results were obtained for the antifungal activity, with Saprolegnia exhibiting higher sensitivity to the synthesized nanoparticles compared to the other species studied. Conclusion: The novel strain Dulcicalothrix alborzica demonstrated potential as a potent producer of AgNPs with unique properties and promising applications in microbial biotechnology.

Sustainable bioconversion of rice straw into indole-3-acetic acid by Streptomyces coelicoflavus using response surface methodology

AbstractRice straw is an abundant agricultural waste that poses environmental disposal challenges and can be utilized for biotechnological applications. This study investigates the potential of actinobacteria to enhance rice straw biodegradation and sustainable indole-3-acetic acid (IAA) production, addressing the need for sustainable agricultural practices. Certain actinobacteria strains can effectively degrade rice straw while optimizing IAA production under controlled fermentation conditions. Twenty actinobacteria isolates were screened for lignocellulolytic enzyme activity, and ten were selected for rice straw biodegradation. IAA production was further optimized using response surface methodology based on temperature, pH, and agitation speed (RPM). Isolate S16 achieved a degradation rate of 68.75%, while S18 produced the highest IAA concentration (1040.625 μg/mL) under optimized conditions (25 °C, pH 9, 160 RPM). The purified IAA significantly improved Medicago sativa L. growth. Furthermore, the 16S rRNA gene sequence of isolate S18 was identified it as Streptomyces coelicoflavus strain NSH24 with accession number PP 320383.1. These findings underscore the potential of actinobacteria to efficiently convert agricultural waste into valuable bioproducts and promote sustainable farming practices. By transforming rice straw into high-value products like IAA, this approach contributes to a circular economy, offering an environmentally friendly solution for biomass utilization and agricultural sustainability.

A New Heuristic Hybrid Model for Cancer-Based P-Glycoprotein Drug Classification: Automated Prediction

As the population improves in real-time applications, drug identification becomes an increasingly difficult task. Various biotechnological applications based on in vitro fertilization can benefit from better drug identification. For many nations, agriculture is the backbone of cancer medication development; increasing crop yields is the key to better drug identification in agricultural contexts. Computer vision applications suffer as a result of the efficient degradation of production quality and quantity caused by p-glycoprotein drugs used in cancer treatment. The identification of cancer-based p-glycoprotein drugs is an aggressive concept in the present day, due to the various symptoms present in cancer. Various authors use soft computing techniques and machine learning approaches to predict these drugs. For the purpose of automatically identifying based p-glycoprotein drugs in cancers, this paper proposes a Hybrid Novel Heuristic Model (HNHM) that combines Self-Organizing Maps (SOMs) with Convolution neural networks (CNNs). The primary challenge in identifying cancer drugs based on p-glycoprotein is feature extraction; hence, dimensionality reduction is crucial in filtering out noise from the cancer sub-cancer drug dataset and separating the subset of the cancer drug dataset affected by the p-glycoprotein drug using support vector machines (SOM). It is a promising approach to automatically identify based p-glycoprotein drugs in cancers by exploring protein features in cancer and predicting which patches are affected by virus-bacteria. CNN is used to match cancer sub-cancer drug data sets. In this study, we use sub-Cancer drug data sets to develop a hybrid heuristic approach to identify based p-glycoprotein drugs found in cancers. Clinical trials of the suggested hybrid model were conducted using sub-cancer drug data sets derived from peach cancers and the publicly available Cancer's drug datasets housed in the UCI repository. With less training required to explore based on p-glycoprotein drugs from sub-Cancer drug datasets, the suggested hybrid model achieves an accuracy of nearly 99.93% when tested on different testing datasets for sub cancer drugs. When compared to other state-of-the-art methods, this one significantly reduces running time while providing efficient performance in a number of p-glycoprotein drug prediction metrics, including recall, sensitivity, and specificity

Compilation of secondary metabolites produced by fungal strains identified as Aspergillus flavus var. oryzae (formerly Aspergillus oryzae), with a review of biotechnological applications, and computational studies of their anti-COVID-19 activity

Compilation of secondary metabolites produced by fungal strains identified as Aspergillus flavus var. oryzae (formerly Aspergillus oryzae), with a review of biotechnological applications, and computational studies of their anti-COVID-19 activity

The Critical Roles of Phosphatidylethanolamine-Binding Proteins in Legumes.

Legumes, characterized by their ability to form symbiotic relationships with nitrogen-fixing bacteria, play crucial roles in agriculture, ecology and human nutrition. Phosphatidylethanolamine-binding proteins (PEBPs) are the key genetic players that contribute to the diverse biological functions of legumes. In this review, we summarize the current understanding of important roles of PEBP genes in legumes, including flowering, inflorescence architecture, seed development and nodulation. We also delve into PEBP regulatory mechanisms and effects on plant growth, development, and adaptation to the environment. Furthermore, we highlight their potential biotechnological applications for crop improvement and promoting sustainable agriculture. This review emphasizes the multifaceted roles of PEBP genes, shedding light on their significance in legume biology and their potential for sustainable productive farming.

Epigenetic factors related to recalcitrance in plant biotechnology

This review explores the challenges and potential solutions in plant micropropagation and biotechnology. While these techniques have proven successful for many species, certain plants or tissues are recalcitrant and do not respond as desired, limiting the application of these technologies due to unattainable or minimal in vitro regeneration rates. Indeed, traditional in vitro culture techniques may fail to induce organogenesis or somatic embryogenesis in some plants, leading to classification as in vitro recalcitrance. This paper focuses on recalcitrance to somatic embryogenesis due to its promise for regenerating juvenile propagules and applications in biotechnology. Specifically, this paper will focus on epigenetic factors that regulate recalcitrance as understanding them may help overcome these barriers. Transformation recalcitrance is also addressed, with strategies proposed to improve transformation frequency. The paper concludes with a review of CRISPR-mediated genome editing's potential in modifying somatic embryogenesis-related epigenetic status and strategies for addressing transformation recalcitrance.