Search

Plant-based biotechnology is crucial for developing renewable biofuels, enhancing bioremediation, and improving the food supply chain. Modern agricultural technologies often face limitations due to variable weather conditions and significant infrastructure costs. Addressing these challenges, smaller-scale biotechnologies optimize plant production through controlled lighting and nutrient supply, mitigating the impact of adverse weather. In vitro technologies are particularly effective for promoting plant growth with desirable phenotypic traits under controlled conditions, though they bring challenges related to sustainability and costs. Recent innovations in microfabrication have led to the miniaturization of traditional in vitro technologies, significantly reducing both sample volumes and costs. This review explores the advancements in microfluidic systems for plant studies, covering a range from single plant cells to mature plants, and discusses the common materials and integrated technologies employed in microfabrication for in-depth in vitro studies. • Small-scale biotechnologies address agricultural problems. • Single-plant cell positioning and observation are possible with microfluidics. • Miniaturized Optimized in vitro conditions improve plant development. • Advanced miniaturized in vitro technology reduced sample volume and cost. • Plant research uses microfluidics for macro and micro inspection. [ABSTRACT FROM AUTHOR]

Physical activity sculpts our bodies, boosts health, and enhances our well-being as we age. But what if your facial skin could also benefit from a similar workout? Imagine achieving such effects through cosmetic products alone. Recent studies have shown the potential for targeted exercises to rejuvenate facial muscles, impacting the skin's internal structure and biological functions, from collagen production to enhancing skin firmness and density. These findings have spurred interest in new methods and technologies for facial toning. Vytrus Biotech has ventured into this innovative field, developing a unique biological synergy designed to strengthen and rejuvenate facial skin by mimicking the natural benefits of physical exercise. This approach enhances appearance and vitality without the need for traditional exercise routines. Using sustainable plant biotechnology, this new cosmetic ingredient activates the skin's exercise-like molecular mechanisms, improving muscle tone for a visibly lifted and rejuvenated complexion. [ABSTRACT FROM AUTHOR]

Grape is one of the most economically significant berry crops. Owing to the biological characteristics of grapes, such as the long juvenile period (5–8 years), high degree of genome heterozygosity, and the frequent occurrence of inbreeding depression, homozygosity during crossbreeding leads to loss of varietal characteristics and viability. CRISPR/Cas editing has become the tool of choice for improving elite technical grape varieties. This study provides the first evidence of a decrease in the total fraction of phenolic compounds and an increase in the concentration of peroxide compounds in grape callus cells upon the addition of chitosan to the culture medium. These previously unreported metabolic features of the grape response to chitosan have been described and used for the first time to increase the probability of selecting plant cells with MLO7 knockout characterised by an oxidative burst in response to the presence of a pathogen modulated by chitosan in the high-metabolite black grape variety 'Merlot'. This was achieved by using a CRISPR/Cas9 editing vector construction with the peroxide sensor HyPer as a reporter. This research represents the first CRISPR/Cas9 editing of 'Merlot', one of the most economically important elite technical grape varieties. [ABSTRACT FROM AUTHOR]

Rutin, a flavonoid phytochemical compound, plays a vital role in human health. It is used in treating capillary fragility and has anti-Alzheimer, anti-inflammatory, and antioxidant effects. In this study, Ti-Mo-Ni-O nanotubes (NTs) were used, for the first time, in an unprecedented plant biotechnology application, wherein in vitro Philodendron shoots (Philodendron erubescens) known as "Imperial Red" were targeted for rutin accumulation. The antioxidant responses and the accumulation of rutin were evaluated in treated Philodendron erubescens (P. erubescens) shoots using 5.0 mg/L of Ti-Mo-Ni-O NTs. The total phenolic content and total flavonoid content were estimated, and an ABTS+ assay, FRAP assay, and iron metal chelation assay were performed. The application of Ti-Mo-Ni-O NTs enhanced the rutin content considerably from 0.02 mg/g to 2.96 mg/g for dry-weight shootlet extracts. Therefore, the use of Ti-Mo-Ni-O NTs is proposed to be a superior alternative to elevate the rutin content. The aim of the current study is to target P. erubescens shoots grown in vitro for the accumulation of rutin compounds using Ti-Mo-Ni-O NT powder, to determine the quantitative and qualitative accumulation of rutin via HPLC–DAD analysis, and to estimate the antioxidant activity of P. erubescens shoot extract. This study presents a novel methodology for utilizing nano-biotechnology in the synthesis of plant secondary metabolites. [ABSTRACT FROM AUTHOR]

Global agricultural production must quadruple by 2050 to fulfil the needs of a growing global population, but climate change exacerbates the difficulty. Cereals are a very important source of food for the world population. Improved cultivars are needed, with better resistance to abiotic stresses like drought, salt, and increasing temperatures, and resilience to biotic stressors like bacterial and fungal infections, and pest infestation. A popular, versatile, and helpful method for functional genomics and crop improvement is genome editing. Rapidly developing genome editing techniques including clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) are very important. This review focuses on how CRISPR/Cas9 genome editing might enhance cereals' agronomic qualities in the face of climate change, providing important insights for future applications. Genome editing efforts should focus on improving characteristics that confer tolerance to conditions exacerbated by climate change (e.g. drought, salt, rising temperatures). Improved water usage efficiency, salt tolerance, and heat stress resilience are all desirable characteristics. Cultivars that are more resilient to insect infestations and a wide range of biotic stressors, such as bacterial and fungal diseases, should be created. Genome editing can precisely target genes linked to disease resistance pathways to strengthen cereals' natural defensive systems. The world population is increasing day by day. Cereals are a good source of energy for this increasing population. Climate changes exert pressure on agriculture and hinder overall production. Genome editing techniques such as CRISPR/Cas have been extensively used to improve crops. In this review, we summarise the different genome editing techniques, their advantages, and disadvantages. This article belongs to the Collection Functional Genomics for Developing Climate Resilient Crops. [ABSTRACT FROM AUTHOR]

Arachis pintoi, commonly known as pinto or forage peanut, is used mainly in consortia with grass pastures and as cover plant. In addition to increasing the productivity of livestock and plantations, it contributes to the mitigation of environmental impacts (reduction of greenhouse gas emissions) and soil improvement (nitrogen fixation, reduction of fertilizers use), as well as to pests and disease management. Several cultivars that are tolerant to specific climates and soil conditions are suitable to be used as ground covers in agroforestry and silvopastoral systems, orchards, and plantations. Biotechnological and phytochemical investigations revealed the potential of pinto peanut as a sustainable source of resveratrol and other stilbenoids. Extracts from plants grown under natural conditions and from materials obtained in vitro displayed allelopathic, anthelmintic, or antioxidant activities. Other studies revealed the potential of pinto peanut for erosion control, phytoremediation, seed and essential oils production, materials for animal tissue engineering, synthesis of nanoparticles for drug delivery, and as green biorefineries to produce proteins, biochemicals, and biomaterials. [ABSTRACT FROM AUTHOR]

This study established in vitro root cultures of Tarenaya atropurpurea from root segments of seedlings and from in vitro propagated plants. Moreover, culture conditions were manipulated aiming to optimize root biomass accumulation and shoot regeneration from newly formed roots was determined. A phytochemical assessment was performed using two extraction methods — dynamic maceration (DM) and ultrasonic assisted extraction (UAE) — and two chromatographic methods for extract analysis (TLC and HPLC). MS medium supplemented with 3.0 mg.L− 1 of indole-3-butyric acid (IBA) induced the highest root multiplication. Root cultures initiated from seedling explants achieved higher biomass accumulation. However, improved root multiplication was achieved using explants from in vitro propagated plants in an optimized culture formulation called Optimum Root Culture Medium (ORCM), which combines MS medium with 1/4 concentration of mineral salts + 3.0 mg.L− 1 IBA + 70 g.L− 1 sucrose, pH 6.5, stirring speed at 130 r.p.m., and 16 h/light. Shoot regeneration from newly formed roots was successfully obtained on MS containing 6-benzylaminopurine (BA). Analysis by TLC suggests the presence of saponins, mainly in root extracts, with the most intense bands acquired by UAE, while HPLC analysis suggests the presence of flavonoids in extracts from aerial parts, with intense signals in extracts obtained by DM. This study was able to establish in vitro root cultures of T. atropurpurea and optimize root biomass accumulation through the manipulation of culture conditions. Phytochemical assessment indicated the presence of saponins and flavonoids, demonstrating potential commercial use of in vitro cultures to produce secondary metabolites in T. atropurpurea.Key message: In vitro root culture of T. atropurpurea was established using root explants from seedlings and in vitro propagated plants. A phytochemical assessment was also performed, applying different extraction and chromatographic methods. [ABSTRACT FROM AUTHOR]

Rapid climate change affects the supply of goods and services needed for a growing world population, reducing crop yields and threatening biodiversity, with potentially catastrophic effects on ecosystems. Particularly at risk are plant species with long life cycles, such as woody species. To address this crisis, there is an urgent need to accelerate breeding pipelines for these species to withstand new environmental conditions and mitigate the effects of climate change. In this article, we highlight examples of genetic engineering in woody species that have the potential to contribute not only to economic development, but also to the conservation and sustainable use of genetic biodiversity, including: the introgression of traits into ecologically important threatened species for large-scale forest restoration, the reduction of air pollution from foliar emissions from bioenergy crops, the acceleration of biomass production to increase CO2 sequestration, the improvement of bioenergy crops to reduce dependence on fossil fuels, and the induction of flowering to reduce the time and resources needed to develop fruit and forest trees varieties with greater resistance to biotic and abiotic stresses. These examples are examined not only for their economic potential but also for their environmental and social impacts. This compilation of biotechnological applications in woody species, aims to enrich the debate on the adoption of genetic engineering in these species. [ABSTRACT FROM AUTHOR]

Matricaria chamomilla (Asteraceae), commonly known as chamomile can tolerate freezing temperatures and grows in many soil types. This plant is found on all continents and has significant medicinal value. There are more than 120 chemicals detected in chamomile flowers, with the majority found in the essential oil. In this study, M. chamomilla was given the NaCl stress of 0 mM, 1 mM, 100 mM, and 150 mM concentrations This study was the first to assess the efficacy of German chamomile upon exposure to salt stress hence plant particles that had been dried and powdered were analyzed using, phytochemical tests, Fourier Transform Infrared and UV–Vis spectroscopy, thin layer chromatography, fluorescence recovery after photobleaching assay, antibacterial and antioxidant activity. The characterization and results of these activities show amazing results which enhance their antibacterial property with an increased zone of inhibition when the samples of salt stress of the above-given concentrations were compared to the control samples. More graph analysis indicates an effective impact of salt stress on the phytoconstituents of M. chamomilla. Other than that, there was a clear flower induction upon salt stress, as a variety of compounds are regarded as essential to the biological functions of chamomile flowers according to the phytoconstituent screening which can be further used in the cosmetic industry, pharmaceutical industry, and all other fields as well for various application as a nano-drug or bio-drug. Due to this, this plant became essential for plant biotechnology research. [ABSTRACT FROM AUTHOR]

Recently, cellular senescence has been of great interest due to its pleiotropic nature and association with both physiological (e.g., aging) and pathological conditions. Excessive accumulation of reactive oxygen species (ROS) can induce inflammation, which accelerates skin aging (also premature aging) and may cause several dermatoses. It has been postulated that plant-derived antioxidants, especially plant stem cells, may prevent cell damage by preserving stemness and reducing cellular senescence by ROS targeting. Therefore, this paper aims to review and summarize recent developments and innovative techniques associated with plant-derived stem cells in relation to skin senescence. This review also presents the possible pathways involved in this process. Particular attention was paid to the potential applications of plant stem cells as senostatics/senomorphics produced by modern plant biotechnology methods. Furthermore, the advantages, limitations, and future directions of this technology are also discussed. This knowledge allows the development of personalized strategies to create a healthy balance in skin care. Plant stem cells could be a more feasible and practical approach to combating the adverse effects of skin senescence processes. [ABSTRACT FROM AUTHOR]

Introducing beneficial productivity and nutritional traits into food, feed and forage crops utilising the tools of biotechnology can lead to improvements via genetic modification that cannot be achieved by traditional plant breeding. The timelines and costs are significant, and the regulatory hurdles can lead to some promising traits failing to be commercialised. These challenges mean that large commodity crops are the primary beneficiaries of biotechnology. New Zealand being primarily a grazing pastoral agricultural market and small in international terms, faces greater challenges. The species used in pastoral agriculture have relatively small seed markets and therefore limited investment for genetic improvement. The nutritional quality of feed and forages both in composition and in energy density per ha has a major influence on animal productivity and the environmental impacts of agriculture. Through government and private industry support, AgResearch has developed novel approaches to improve plant photosynthesis, energy density, and nutritional quality that has applications in crops globally. This review discusses the different challenges and solutions to improving plant nutrient density and outlines the benefits of these novel biotechnology traits in animal forage and feed crops. [ABSTRACT FROM AUTHOR]

Potato is a very important crop for human nutrition worldwide. Potato disease and pests can cause economic yield losses. In this study, approximately 130.000 potato seeds from different genetic backgrounds were obtained, evaluated, and selected from 2008 to 2016 for developing new cultivars. After several years, superior lines were tested in different locations to select new cultivars. At the end of the potato breeding program, eight superior potato lines (Nos 12-55-07, 12-55-16, 12-68-05, 12-69-39, 13-67-25, 13-66-23, 12-45-24 and 13-66-75) were submitted for registration as commercial cultivars. The use of resistance genes is the most effective method for controlling these diseases and pests, and DNA markers that are tightly linked to resistance genes are available. The aim of this research was to evaluate the use of closely linked molecular markers for combining Gro1 and H1 for cist nematode resistance, Sen1 for wart resistance or with R1 for late blight or with Rx1 for PVX in advanced breeding lines and commercial candidate cultivars. In the research, 61 advanced lines were checked for the presence of five markers. As a result, only two breeding lines (Nos 31-01-03 and 32-02-52) were determined to have positive results for four markers, and nine advanced lines were found to have positive marker results for Sen1, H1, and R1 at the same time. In 6 advanced lines, positive results were acquired from both Sen1, H1, and Rx1, 2 lines Sen1, Gro1, and H1, and only one line No. 12-45-24 Sen1, Gro1, H1, and Rx1 were found to have positive results for markers. Marker-assisted selection for cist nematode, wart, late blight, and PVX will be performed using potato breeding programs. [ABSTRACT FROM AUTHOR]

Somatic embryogenesis (SE) is a biotechnological tool used to generate new individuals and is the preferred method for rapid plant regeneration. However, the molecular basis underlying somatic cell regeneration through SE is not yet fully understood, particularly regarding interactions between the proteome and post-translational modifications. Here, we performed association analysis of high-throughput proteomics and phosphoproteomics in three representative samples (non-embryogenic calli, NEC; primary embryogenic calli, PEC; globular embryos, GE) during the initiation of plant regeneration in cotton, a pioneer crop for genetic biotechnology applications. Our results showed that protein accumulation is positively regulated by phosphorylation during SE, as revealed by correlation analyses. Of the 1418 proteins that were differentially accumulated in the proteome and the 1106 phosphoproteins that were differentially regulated in the phosphoproteome, 115 proteins with 229 phosphorylation sites overlapped (co-differential). Furthermore, seven dynamic trajectory patterns of differentially accumulated proteins (DAPs) and the correlated differentially regulated phosphoproteins (DRPPs) pairs with enrichment features were observed. During the initiation of plant regeneration, functional enrichment analysis revealed that the overlapping proteins (DAPs-DRPPs) were considerably enriched in cellular nitrogen metabolism, spliceosome formation, and reproductive structure development. Moreover, 198 DRPPs (387 phosphorylation sites) were specifically regulated at the phosphorylation level and showed four patterns of stage-enriched phosphorylation susceptibility. Furthermore, enrichment annotation analysis revealed that these phosphoproteins were significantly enriched in endosomal transport and nucleus organization processes. During embryogenic differentiation, we identified five DAPs-DRPPs with significantly enriched characteristic patterns. These proteins may play essential roles in transcriptional regulation and signaling events that initiate plant regeneration through protein accumulation and/or phosphorylation modification. This study enriched the understanding of key proteins and their correlated phosphorylation patterns during plant regeneration, and also provided a reference for improving plant regeneration efficiency. [ABSTRACT FROM AUTHOR]

Background: Kale, a versatile cruciferous crop, valued for its pro-health benefits, stress resistance, and potential applications in forage and cosmetics, holds promise for further enhancement of its bioactive compounds through in vitro cultivation methods. Micropropagation techniques use cytokinins (CKs) which are characterized by various proliferative efficiency. Despite the extensive knowledge regarding CKs, there remains a gap in understanding their role in the physiological mechanisms. That is why, here we investigated the effects of three CKs – kinetin (Kin), 6-benzylaminopurine (BAP), and 2-isopentenyladenine (2iP) – on kale physiology, antioxidant status, steroidal metabolism, and membrane integrity under in vitro cultivation. Results: Our study revealed that while BAP and 2iP stimulated shoot proliferation, they concurrently diminished pigment levels and photosynthetic efficiency. Heightened metabolic activity in response to all CKs was reflected by increased respiratory rate. Despite the differential burst of ROS, the antioxidant properties of kale were associated with the upregulation of guaiacol peroxidase and the scavenging properties of ascorbate rather than glutathione. Notably, CKs fostered the synthesis of sterols, particularly sitosterol, pivotal for cell proliferation and structure of membranes which are strongly disrupted under the action of BAP and 2iP possibly via pathway related to phospholipase D and lipoxygenase which were upregulated. Intriguingly, both CKs treatment spurred the accumulation of sitostenone, known for its ROS scavenging and therapeutic potential. The differential effects of CKs on brassicasterol levels and brassinosteroid (BRs) receptor suggest potential interactions between CKs and BRs. Conclusion: Based on the presented results we conclude that the effect evoked by BAP and 2iP in vitro can improve the industrial significance of kale because this treatment makes possible to control proliferation and/or biosynthesis routes of valuable beneficial compounds. Our work offers significant insights into the nuanced effects of CKs on kale physiology and metabolism, illuminating potential avenues for their application in plant biotechnology and medicinal research. [ABSTRACT FROM AUTHOR]

Background: Several members of the Lamiaceae family of plants produce large amounts of essential oil [EO] that find extensive applications in the food, cosmetics, personal hygiene, and alternative medicine industries. There is interest in enhancing EO metabolism in these plants. Main body: Lavender produces a valuable EO that is highly enriched in monoterpenes, the C10 class of the isoprenoids or terpenoids. In recent years, substantial effort has been made by researchers to study terpene metabolism and enhance lavender EO through plant biotechnology. This paper reviews recent advances related to the cloning of lavender monoterpene biosynthetic genes and metabolic engineering attempts aimed at improving the production of lavender monoterpenes in plants and microbes. Conclusion: Metabolic engineering has led to the improvement of EO quality and yield in several plants, including lavender. Furthermore, several biologically active EO constituents have been produced in microorganisms. [ABSTRACT FROM AUTHOR]

Pseudomonas bacteria are renowned for their remarkable capacity to synthesize antibiotics, namely mupirocin, gluconic acid, pyrrolnitrin, and 2,4-diacetylphloroglucinol (DAPG). While these substances are extensively employed in agricultural biotechnology to safeguard plants against harmful bacteria and fungi, their potential for human medicine and healthcare remains highly promising for common science. However, the challenge of obtaining stable producers that yield higher quantities of these antibiotics continues to be a pertinent concern in modern biotechnology. Although the interest in antibiotics of Pseudomonas bacteria has persisted over the past century, many uncertainties still surround the regulation of the biosynthetic pathways of these compounds. Thus, the present review comprehensively studies the genetic organization and regulation of the biosynthesis of these antibiotics and provides a comprehensive summary of the genetic organization of antibiotic biosynthesis pathways in pseudomonas strains, appealing to both molecular biologists and biotechnologists. In addition, attention is also paid to the application of antibiotics in plant protection. [ABSTRACT FROM AUTHOR]

This study investigates the effects of artificial in vitro polyploidisation of Astragalus membranaceus, focusing specifically on gene expression and metabolic pathway associated with the biosynthesis of calycosin and calycosin-7-O-β-D-glucoside. Using oryzalin as an antimitotic agent, we have systematically investigated different genotypic lines, under both in vitro and ex vitro conditions. Amid cases of reduced gene expression in certain lines, results showed a significant upregulation in specific lines, particularly in genotypes 16, 54, and 74. Genotype 54 showed an exceptional response, manifesting a statistically significant upregulation in all investigated genes studied under in vitro conditions (i.e. AmPAL, AmC4H and AmI3′H). Interestingly, even under ex vitro conditions after two years of greenhouse cultivation, certain lines showed significant variations in gene expression. The genotype 16, although no longer tetraploid, exhibited the highest expression levels among the examined genotypes, with statistically significant upregulation of both the AmPAL and AmC4H genes. In addition, the induced autotetraploid genotype 74 showed a significant upregulation of the AmI3′H gene and a concomitant downregulation of the AmC4H gene. These results highlight the complex regulatory mechanisms affected by the polyploidisation of A. membranaceus and provide promising avenues for manipulating gene expression to enhance the production of pharmacologically significant compounds. Key message: This study investigates the effects of artificial polyploidisation on gene expression of Astragalus membranaceus, suggesting diverse regulatory influences for potential plant bioactive compound production. [ABSTRACT FROM AUTHOR]

The primary focus of medicinal cannabis research is to ensure the stability of cannabis lines for consistent administration of chemically uniform products to patients. In recent years, tissue culture has emerged as a valuable technique for genetic preservation and rapid multiplication of cannabis clones. However, there is concern that the physical and chemical conditions of the growing media can induce somaclonal variation, potentially impacting the viability and uniformity of clones. To address this concern, we developed Comparative Restriction Enzyme Analysis of Methylation (CREAM), a novel method to assess DNA methylation patterns and used it to study a population of 78 cannabis clones maintained in tissue culture. Through bioinformatics analysis of the methylome, we successfully detected 2,272 polymorphic methylated regions among the clones. Remarkably, our results demonstrated that DNA methylation patterns were preserved across subcultures within the clonal population, allowing us to distinguish between two subsets of clonal lines used in this study. These findings significantly contribute to our understanding of the epigenetic variability within clonal lines in medicinal cannabis produced through tissue culture techniques. This knowledge is crucial for understanding the effects of tissue culture on DNA methylation and ensuring the consistency and reliability of medicinal cannabis products with therapeutic properties. Additionally, the CREAM method is a fast and affordable technology to get a first glimpse at methylation in a biological system. It offers a valuable tool for studying epigenetic variation in other plant species, thereby facilitating broader applications in plant biotechnology and crop improvement. [ABSTRACT FROM AUTHOR]

Food production and food security depend on the ability of crops to cope with anthropogenic climate change and successfully produce seed. To guarantee food production well into the future, contemporary plant scientists in Latin America must carry out research on how plants respond to environmental stressors such as temperature, drought, and salinity. This review shows the opportunities to apply these results locally and abroad and points to the gaps that still exist in terms of reproductive processes with the purpose to better link research with translational work in plant breeding and biotechnology. Suggestions are put forth to address these gaps creatively in the face of chronic low investment in science with a focus on applicability. [ABSTRACT FROM AUTHOR]

Abstract Background Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), belonging to ω-3 long-chain polyunsaturated fatty acids (ω3-LC-PUFAs), are essential components of human diet. They are mainly supplemented by marine fish consumption, although their native producers are oleaginous microalgae. Currently, increasing demand for fish oils is insufficient to meet the entire global needs, which puts pressure on searching for the alternative solutions. One possibility may be metabolic engineering of plants with an introduced enzymatic pathway producing ω3-LC-PUFAs. Result In this study we focused on the acyl-CoA:diacylglycerol acyltransferase2b (PtDGAT2b) from the diatom Phaeodactylum tricornutum, an enzyme responsible for triacylglycerol (TAG) biosynthesis via acyl-CoA-dependent pathway. Gene encoding PtDGAT2b, incorporated into TAG-deficient yeast strain H1246, was used to confirm its activity and conduct biochemical characterization. PtDGAT2b exhibited a broad acyl-CoA preference with both di-16:0-DAG and di-18:1-DAG, whereas di-18:1-DAG was favored. The highest preference for acyl donors was observed for 16:1-, 10:0- and 12:0-CoA. PtDGAT2b also very efficiently utilized CoA-conjugated ω-3 LC-PUFAs (stearidonic acid, eicosatetraenoic acid and EPA). Additionally, verification of the potential role of PtDGAT2b in planta, through its transient expression in tobacco leaves, indicated increased TAG production with its relative amount increasing to 8%. Its co-expression with the gene combinations aimed at EPA biosynthesis led to, beside elevated TAG accumulation, efficient accumulation of EPA which constituted even 25.1% of synthesized non-native fatty acids (9.2% of all fatty acids in TAG pool). Conclusions This set of experiments provides a comprehensive biochemical characterization of DGAT enzyme from marine microalgae. Additionally, this study elucidates that PtDGAT2b can be used successfully in metabolic engineering of plants designed to obtain a boosted TAG level, enriched not only in ω-3 LC-PUFAs but also in medium-chain and ω-7 fatty acids.

As photosynthetic microorganisms, cyanobacteria play a dominant part in numerous ecological systems owing to their ability to fix carbon and nitrogen and are therefore an essential part of primary production in both aquatic and terrestrial environments. The utility of nitrogen-fixing cyanobacteria in plant biotechnology opens up promising strategies for the conservation and sustainable use of rare, endangered plant species and bioactive cell cultures. Here, we discuss the complicated physiological aspects of biological nitrogen fixation in cyanobacteria and their symbiotic relationship with plants. This review focuses on recent advances in biotechnological tools such as CRISPR-Cas9, nanotechnology and multiomics-based approaches for enhancing plant regeneration systems to cultivate specialized metabolites. We also look at the methods in vitro preservation of plants and how to scale up a culture using bioreactor systems. The review ends by highlighting the promise of cyanobacteria-powered plant biotechnology as a renewable mechanism for rare species conservation and specialized metabolites production, providing an optimistic modal, formative future direction in plant biosynthesis.

Schisandra henryi is an endemic species of medicinal potential known from traditional Chinese medicine. As part of this study, a complex biotechnological and phytochemical assessment was conducted on S. henryi with a focus on phenolic compounds and antioxidant profiling. The following in vitro cultures were tested: microshoot agar and callus, microshoot agitated, and suspension, along with the microshoot culture in PlantForm bioreactors. Qualitative profiling was performed by ultra-high-performance liquid chromatography with a photodiode array detector coupled with ion-trap mass spectrophotometry with electrospray ionization and then quantitative analysis by high-performance liquid chromatography with a diode array detector using standards. In the extracts, mainly the compounds from procyanidins were identified as well as phenolic acids (neochlorogenic acid, caffeic acid, protocatechuic acid) and catechin. The highest content of phenolic compounds was found for in vitro agar microshoot culture (max. total content 229.87 mg/100 g DW) and agitated culture (max. total content 22.82 mg/100 g DW). The max. TPC measured using the Folin-Ciocalteu assay was equal to 1240.51 mg GAE/100 g DW (agar microshoot culture). The extracts were evaluated for their antioxidant potential by the DPPH, FRAP, and chelate iron ion assays. The highest potential was indicated for agar microshoot culture (90% of inhibition and 59.31 nM/L TEAC, respectively). The research conducted on the polyphenol profiling and antioxidant potential of S. henryi in vitro culture extracts indicates the high therapeutic potential of this species. Key points: • Different types of S. henryi in vitro cultures were compared for the first time. • The S. henryi in vitro culture strong antioxidant potential was determined for the first time. • The polyphenol profiling of different types of S. henryi in vitro cultures was shown. [ABSTRACT FROM AUTHOR]

Summary: Somatic cell totipotency in plant regeneration represents the forefront of the compelling scientific puzzles and one of the most challenging problems in biology. How somatic embryogenic competence is achieved in regeneration remains elusive.Here, we discover uncharacterized organelle‐based embryogenic differentiation processes of intracellular acquisition and intercellular transformation, and demonstrate the underlying regulatory system of somatic embryogenesis‐associated lipid transfer protein (SELTP) and its interactor calmodulin1 (CAM1) in cotton as the pioneer crop for biotechnology application. The synergistic CAM1 and SELTP exhibit consistent dynamical amyloplast–plasmodesmata (PD) localization patterns but show opposite functional effects.CAM1 inhibits the effect of SELTP to regulate embryogenic differentiation for plant regeneration. It is noteworthy that callus grafting assay reflects intercellular trafficking of CAM1 through PD for embryogenic transformation.This work originally provides insight into the mechanisms responsible for embryogenic competence acquisition and transformation mediated by the Ca2+/CAM1‐SELTP regulatory pathway, suggesting a principle for plant regeneration and cell/genetic engineering. [ABSTRACT FROM AUTHOR]

Summary: Transient expression in Nicotiana benthamiana offers a robust platform for the rapid production of complex secondary metabolites. It has proven highly effective in helping identify genes associated with pathways responsible for synthesizing various valuable natural compounds. While this approach has seen considerable success, it has yet to be applied to uncovering genes involved in anthocyanin biosynthetic pathways. This is because only a single anthocyanin, delphinidin 3‐O‐rutinoside, can be produced in N. benthamiana by activation of anthocyanin biosynthesis using transcription factors. The production of other anthocyanins would necessitate the suppression of certain endogenous flavonoid biosynthesis genes while transiently expressing others. In this work, we present a series of tools for the reconstitution of anthocyanin biosynthetic pathways in N. benthamiana leaves. These tools include constructs for the expression or silencing of anthocyanin biosynthetic genes and a mutant N. benthamiana line generated using CRISPR. By infiltration of defined sets of constructs, the basic anthocyanins pelargonidin 3‐O‐glucoside, cyanidin 3‐O‐glucoside and delphinidin 3‐O‐glucoside could be obtained in high amounts in a few days. Additionally, co‐infiltration of supplementary pathway genes enabled the synthesis of more complex anthocyanins. These tools should be useful to identify genes involved in the biosynthesis of complex anthocyanins. They also make it possible to produce novel anthocyanins not found in nature. As an example, we reconstituted the pathway for biosynthesis of Arabidopsis anthocyanin A5, a cyanidin derivative and achieved the biosynthesis of the pelargonidin and delphinidin variants of A5, pelargonidin A5 and delphinidin A5. [ABSTRACT FROM AUTHOR]

Background: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), belonging to ω-3 long-chain polyunsaturated fatty acids (ω3-LC-PUFAs), are essential components of human diet. They are mainly supplemented by marine fish consumption, although their native producers are oleaginous microalgae. Currently, increasing demand for fish oils is insufficient to meet the entire global needs, which puts pressure on searching for the alternative solutions. One possibility may be metabolic engineering of plants with an introduced enzymatic pathway producing ω3-LC-PUFAs. Result: In this study we focused on the acyl-CoA:diacylglycerol acyltransferase2b (PtDGAT2b) from the diatom Phaeodactylum tricornutum, an enzyme responsible for triacylglycerol (TAG) biosynthesis via acyl-CoA-dependent pathway. Gene encoding PtDGAT2b, incorporated into TAG-deficient yeast strain H1246, was used to confirm its activity and conduct biochemical characterization. PtDGAT2b exhibited a broad acyl-CoA preference with both di-16:0-DAG and di-18:1-DAG, whereas di-18:1-DAG was favored. The highest preference for acyl donors was observed for 16:1-, 10:0- and 12:0-CoA. PtDGAT2b also very efficiently utilized CoA-conjugated ω-3 LC-PUFAs (stearidonic acid, eicosatetraenoic acid and EPA). Additionally, verification of the potential role of PtDGAT2b in planta, through its transient expression in tobacco leaves, indicated increased TAG production with its relative amount increasing to 8%. Its co-expression with the gene combinations aimed at EPA biosynthesis led to, beside elevated TAG accumulation, efficient accumulation of EPA which constituted even 25.1% of synthesized non-native fatty acids (9.2% of all fatty acids in TAG pool). Conclusions: This set of experiments provides a comprehensive biochemical characterization of DGAT enzyme from marine microalgae. Additionally, this study elucidates that PtDGAT2b can be used successfully in metabolic engineering of plants designed to obtain a boosted TAG level, enriched not only in ω-3 LC-PUFAs but also in medium-chain and ω-7 fatty acids. [ABSTRACT FROM AUTHOR]

Phytosulfokine-a (PSK-a) is a disulfated pentapeptide (YIYTQ) acting as an intercellular signal peptide and growth factor. It was originally isolated from conditioned medium of asparagus mesophyll cell cultures in 1996 and later characterized as a hormone-like signal molecule with important roles in numerous processes of in vivo plant growth and development. It is currently becoming a valuable mitogenic factor in plant breeding and biotechnology due to its stimulatory effect on in vitro cell elongation, proliferation and differentiation. The focus of our work was to review current knowledge about the roles of PSK-a in plant biotechnology and to evaluate its influence on the regeneration of protoplasts of four Brassica oleracea cultivars (two cauliflower and two cabbage) cultured under two distinctive protocols and with different protoplast densities. Protoplast regeneration was studied due to its high value for plant genome editing, which is generally limited by the inefficient regeneration of treated protoplasts of numerous important plant genotypes. Our hypothesis was that the stress related to PEG-mediated protoplast transformation and the following decrease in viable protoplast density in culture could be alleviated by the addition of PSK-a to the culture medium. We therefore tested whether PSK-a could increase cell division at the early stages of culture (5 and 15 days after protoplast isolation) and stimulate the formation of microcallus colonies up to the 30st day of culture and to evaluate its influence on callus organogenesis leading to shoot regeneration. The PSK-a showed a strong stimulatory effect on untransformed protoplast regeneration already during the first days of culture, accelerating cell division up to 5.3-fold and the formation of multicellular microcallus colonies up to 37.0-fold. The beneficial influence was retained at later stages of regeneration, when PSK improved shoot organogenesis even if it was present only during the first 10 days of culture. The highest numbers of shoots, however, were regenerated when PSK was present during the first days of culture and later in solid shoot regeneration medium. Finally, the addition of PSKa to PEG-transformed protoplasts significantly enhanced their division rate and the formation of microcallus colonies in selection media, up to 44.0-fold. [ABSTRACT FROM AUTHOR]

Yellow nutsedge (Cyperus esculentus L.) is cultivated worldwide due to its agricultural and biotechnological potential. In Brazil, it is considered a weed, and we lack studies on its cultivation. Overcoming tuber dormancy is crucial for propagation. This study aimed to assess various dormancy-breaking methods' effects on tubers and initial plant development. The treatments included gibberellic acid immersion, ethylene exposure, purple nutsedge extract immersion, temperature conditioning, scarification, and bud cutting, along with a control. Scarification resulted in the shortest emergence time (0.904 days) and fastest emergence speed (5.092 tubers/day). Plant development was minimally affected by the treatments, with scarification and gibberellic acid (100 mg L−1) resulting in taller plants (1.19–1.23 times higher than the control). The conditioning at 4 °C and 70 °C proved to be less effective in breaking dormancy. Purple nutsedge extract immersion and bud cutting hindered plant growth. Scarification emerged as the most effective dormancy-breaking method. This study provides insights into the cultivation of yellow nutsedge in Brazil, highlighting the effectiveness of scarification in improving tuber germination and the early growth stages of plants. [ABSTRACT FROM AUTHOR]

This research's scope encompassed biotechnological, phytochemical, and biological studies of Schisandra henryi, including investigations into its in vitro microshoot culture grown in PlantForm bioreactors (temporary immersion systems, TISs), as well as extracts from leaves of the parent plant, focusing on anti-inflammatory, antioxidant, anticancer, and antimicrobial activities. The phytochemical analysis included the isolation and quantification of 17 compounds from dibenzocyclooctadiene, aryltetralin lignans, and neolignans using centrifugal partition chromatography (CPC), HPLC-DAD, and UHPLC-MS/MS tandem mass spectrometry with triple quadrupole mass filter methods. Higher contents of compounds were found in microshoots extracts (max. 543.99 mg/100 g DW). The major compound was schisantherin B both in the extracts from microshoots and the leaves (390.16 and 361.24 mg/100 g DW, respectively). The results of the anti-inflammatory activity in terms of the inhibition of COX-1, COX-2, sPLA2, and LOX-15 enzymes indicated that PlantForm microshoot extracts showed strong activity against COX-1 and COX-2 (for 177 mg/mL the inhibition percentage was 76% and 66%, respectively). The antioxidant potential assessed using FRAP, CUPRAC, and DPPH assays showed that extracts from microshoot cultures had 5.6, 3.8, and 3.3 times higher power compared to extracts from the leaves of the parent plant, respectively. The total polyphenol content (TPC) was 4.1 times higher in extracts from the in vitro culture compared to the leaves. The antiproliferative activity against T-cell lymphoblast line Jurkat, breast adenocarcinoma cultures (MCF-7), colon adenocarcinoma (HT-29), and cervical adenocarcinoma (HeLa), showed that both extracts have considerable effects on the tested cell lines. The antimicrobial activity tested against strains of Gram-positive and Gram-negative bacteria and fungi showed the highest activity towards H. pylori (MIC and MBC 0.625 mg/mL). [ABSTRACT FROM AUTHOR]

Nanocarriers protect RNAs from enzymatic degradation after application onto plants for sustained protection against viruses and insects. Nanocarriers enabled controlled delivery of nucleic acids to plant cells, including slow and sustained release, subcellular organ targeting delivery, stimulated release, and endosome escape. The nanoparticle-mediated delivery system has high versatility, in terms of applicability to all plant species and the capacity to deliver alternative forms of nucleic acids and proteins. Plant biotechnology plays a crucial role in developing modern agriculture and plant science research. However, the delivery of exogenous genetic material into plants has been a long-standing obstacle. Nanoparticle-based delivery systems are being established to address this limitation and are proving to be a feasible, versatile, and efficient approach to facilitate the internalization of functional RNA and DNA by plants. The nanoparticle-based delivery systems can also be designed for subcellular delivery and controlled release of the biomolecular cargo. In this review, we provide a concise overview of the recent advances in nanocarriers for the delivery of biomolecules into plants, with a specific focus on applications to enhance RNA interference, foreign gene transfer, and genome editing in plants. [ABSTRACT FROM AUTHOR]

Introduction: Panax vietnamensis is a valuable medicinal plant and a source of a broad spectrum of biologically active ginsenosides of different structural groups. Overexploitation and low adaptability to planation cultivation have made this species vulnerable to human pressure and prompted the development of cell cultivation in vitro as a sustainable alternative to harvesting wild plants for their bioactive components. Despite high interest in biotechnological production, little is known about themain factors affecting cell growth and ginsenoside biosynthesis of this species under in vitro conditions. In this study, the potential of cell cultures of P. vietnamensis as a biotechnological source of ginsenosides was was assessed. Methods: Six suspension cell lines that were developed from different sections of a single rhizome through a multi-step culture optimization process and maintained for over 3 years on media with different mineral salt base and varying contents of auxins and cytokinins. These cell lines were evaluated for productivity parameters and cytological characteristics. Ginsenoside profiles were assessed using a combination of the reversed-phase ultra-high-performance liquid chromatography-Orbitrap-tandem mass spectrometry (UHPLC-Orbitrap-MS/MS) and ultra-performance liquid chromatography-time of flight-mass spectrometry (UPLC-TOF-MS). Results: All lines demonstrated good growth with a specific growth rate of 0.1-0.2 day-1, economic coefficient of 0.31-0.70, productivity on dry weight (DW) of 0.30-0.83 gDW (L·day)-1, and maximum biomass accumulation varying from 10 to 22 gDW L-1. Ginsenosides of the protopanaxadiol (Rb1, Rb2/Rb3, malonyl-Rb1, and malonyl-Rb2/Rb3), oleanolic acid (R0 and chikusetsusaponin IV), and ocotillol (vinaginsenoside R1) groups and their isomers were identified in cell biomass extracts. Chikusetsusaponin IV was identified in P. vietnamensis cell culture for the first time. Discussion: These results suggest that suspension cell cultures of Vietnamese ginseng have a high potential for the biotechnological production of biomass containing ginsenosides, particularly of the oleanolic acid and ocotillol groups. [ABSTRACT FROM AUTHOR]

Plant proteases are essential enzymes that play key roles during crucial phases of plant life. Some proteases are mainly involved in general protein turnover and recycle amino acids for protein synthesis. Other proteases are involved in cell signalling, cleave specific substrates and are key players during important genetically controlled molecular processes. Cathepsin B is a cysteine protease that can do both because of its exopeptidase and endopeptidase activities. Animal cathepsin B has been investigated for many years, and much is known about its mode of action and substrate preferences, but much remains to be discovered about this potent protease in plants. Cathepsin B is involved in plant development, germination, senescence, microspore embryogenesis, pathogen defence and responses to abiotic stress, including programmed cell death. This review discusses the structural features, the activity of the enzyme and the differences between the plant and animal forms. We discuss its maturation and subcellular localisation and provide a detailed overview of the involvement of cathepsin B in important plant life processes. A greater understanding of the cell signalling processes involving cathepsin B is needed for applied discoveries in plant biotechnology. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) is a powerful tool for understanding and manipulating signaling pathways in plant science, potentially facilitating the accelerated development of novel plant traits and crop yield improvement. The common strategy for delivering siRNA into intact plants using agrobacterium or viruses is complicated and time‐consuming, limiting the application of RNAi in plant research. Here, a novel delivery method based on mesoporous silica nanoparticles (MSNs) is reported, which allows for the efficient delivery of siRNA into mature plant leaves via topical application without the aid of mechanical forces, achieving transient gene knockdown with up to 98% silencing efficiency at the molecular level. In addition, this method is nontoxic to plant leaves, enabling the repeated delivery of siRNA for long‐term silencing. White spots and yellowing phenotypes are observed after spraying the MSN‐siRNA complex targeted at phytoene desaturase and magnesium chelatase genes. After high light treatment, photobleaching phenotypes are also observed by spraying MSNs‐siRNA targeted at genes into the Photosystem II repair cycle. Furthermore, the study demonstrated that MSNs can simultaneously silence multiple genes. The results suggest that MSN‐mediated siRNA delivery is an effective tool for long‐term multi‐gene silencing, with great potential for application in plant functional genomic analyses and crop improvement. [ABSTRACT FROM AUTHOR]

Key message: Single-cell transcriptomic techniques have emerged as powerful tools in plant biology, offering high-resolution insights into gene expression at the individual cell level. This review highlights the rapid expansion of single-cell technologies in plants, their potential in understanding plant development, and their role in advancing plant biotechnology research. Single-cell techniques have emerged as powerful tools to enhance our understanding of biological systems, providing high-resolution transcriptomic analysis at the single-cell level. In plant biology, the adoption of single-cell transcriptomics has seen rapid expansion of available technologies and applications. This review article focuses on the latest advancements in the field of single-cell transcriptomic in plants and discusses the potential role of these approaches in plant development and expediting plant biotechnology research in the near future. Furthermore, inherent challenges and limitations of single-cell technology are critically examined to overcome them and enhance our knowledge and understanding. [ABSTRACT FROM AUTHOR]

The water hyacinth has garnered significant attention due to its harm effect on economics and ecology. Thus, understanding Echhornia crassipes at different levels is necessary to figure out the suitable methods either for withstanding or using this species positively. Eichhornia crassipes is a synonym of Pontederia crassipes, a fast-growing hydrophyte, native to South America, belonging to genus Pontederia and family Pontederiaceae, monocots. The anatomy of different organs was addressed to provide a clear image of the internal structure of the species under study. This could explain the survival of this plant in the sewage ecosystem. Aerinchyma tissue was found in all organs, and water was obviously stored in the root to balance plant floating. In addition, pollen grains' characterization was recognized on SEM stubs with their monads, heteropoler, radiosymetry and large size (P-56/E-45µm). Identification of Echhornia crassipes was approved via PCR and partial gene sequencing of rbcL gene (Ribulose Bisphosphate Carboxylase), which coincides with the gen bank. The present study encompasses molecular identification, as well as an anatomical and paleontological illustration. [ABSTRACT FROM AUTHOR]

Lignin, a complex aromatic polymer, is considered a part of the secondary cell wall. Also, it's called a "cell adhesive" based on provides strength and rigidity to plant tissues and fibers through strong cross-linking with cell wall components. Moreover, lignin acts as a line of defense for plants against biotic and abiotic stresses. In the field of plant biotechnology, particle bombardment remains the most important technique to directly transfer DNA from source to target organ and the most widely used method to generate transgenic plants for commercial purposes. Down-regulation of Jatropha curcas's lignin-producing caffeic acid 3-O-methyltransferase (COMT) gene production pathway led the transgenic plants to develop cell walls. The transgene COMT antisense is used as a selection marker pZMAS-COMT 10,907 bp plasmid (University of Illinois) to confer resistance to glufosinate herbicides under the 35S promoter. Irradiation of embryonic callus with bullet particles introduced the COMT antisense gene and reduced the amount of lignin. Transgenic green callus was used to extract DNA, RNA, and PCR & RT-PCR were performed to detect the presence of genes. A positive sharp band appeared in the green callus, indicating that the gene expression level was downregulated compared to the control. Transgenic shoots and green callus had lower lignin content compared to controls. [ABSTRACT FROM AUTHOR]

The endoribonuclease RNase P is responsible for tRNA 50 maturation in all domains of life. A unique feature of RNase P is the variety of enzyme architectures, ranging from dual- to multi-subunit ribonucleoprotein forms with catalytic RNA subunits to protein-only enzymes, the latter occurring as singleor multi-subunit forms or homo-oligomeric assemblies. The protein-only enzymes evolved twice: a eukaryal protein-only RNase P termed PRORP and a bacterial/archaeal variant termed homolog of Aquifex RNase P (HARP); the latter replaced the RNA-based enzyme in a small group of thermophilic bacteria but otherwise coexists with the ribonucleoprotein enzyme in a few other bacteria as well as in those archaea that also encode a HARP. Here we summarize the history of the discovery of protein-only RNase P enzymes and review the state of knowledge on structure and function of bacterial HARPs and eukaryal PRORPs, including human mitochondrial RNase P as a paradigm of multi-subunit PRORPs. We also describe the phylogenetic distribution and evolution of PRORPs, as well as possible reasons for the spread of PRORPs in the eukaryal tree and for the recruitment of two additional protein subunits to metazoan mitochondrial PRORP. We outline potential applications of PRORPs in plant biotechnology and address diseases associated with mutations in human mitochondrial RNase P genes. Finally, we consider possible causes underlying the displacement of the ancient RNA enzyme by a protein-only enzyme in a small group of bacteria. [ABSTRACT FROM AUTHOR]

Single-cell approaches are a promising way to obtain high-resolution transcriptomics data and have the potential to revolutionize the study of plant growth and development. Recent years have seen the advent of unprecedented technological advances in the field of plant biology to study the transcriptional information of individual cells by single-cell RNA sequencing (scRNA-seq). This review focuses on the modern advancements of single-cell transcriptomics in plants over the past few years. In addition, it also offers a new insight of how these emerging methods will expedite advance research in plant biotechnology in the near future. Lastly, the various technological hurdles and inherent limitations of single-cell technology that need to be conquered to develop such outstanding possible knowledge gain is critically analyzed and discussed. [ABSTRACT FROM AUTHOR]

Amoebiasis is an intestinal disease caused by a unicellular parasite called Entamoeba histolytica. Interleukin-35 (IL-35) is the youngest specific member of the IL-12 family that plays a major role in the inhibitory function of regulatory T cells (Tregs) to curb inflammatory responses. IL-25 of the IL-17 family, which is widely released by Th2 cells and epithelial cells, is a warning signal produced upon cell or tissue injury to activate immune cells. The present study aimed to determine the cytokine profile (IL-25 and IL-35) in patients with E. histolytica infection in southern Iraq. This hospital-based study was conducted from August 2022 to May 2023. The study participants were patients with E. histolytica infection admitted to the infection department of general hospitals in Thi-qar province, southern Iraq. Initially, E. histolytica amebiasis was detected in the patients by nested multiplex PCR. All collected sera were tested with the Human Interleukin 35 (Biotech, China, Cat.RD-IL35-Hu) and IL25 (Biotech, China, Cat.RD-IL25-Hu) ELISA kits according to the instructions of the manufacturer. A total of 80 patients, including 50 patients with E. histolytica infection and 30 subjects in the control group without E. histolytica infection, were enrolled in the present study. The results showed a significant difference (p<0.001) in the serum level of IL-25 in patients with E. histolytica infection (4275.19 pg/mL) compared to individuals in the control group without E. histolytica infection (2186 pg/mL). Statistical analysis showed that there was no significant difference in the serum levels of IL-35 patients with E. histolytica infection compared with individuals in the control group without E. histolytica infection. The results of the present study show that the level of IL-25 is high in patients with E. histolytica infection. This indicates the important role of IL-25 in the activation of the immune system during intestinal inflammation. Therefore, this cytokine can be used as a diagnostic marker for E. histolytica infection. [ABSTRACT FROM AUTHOR]

The medicinal use of mountain arnica (Arnica montana L.) has a long tradition in Europe and is currently confirmed by the documented biological activity of extracts from its flowers and helenalin isolated from them. The main groups of specialized metabolites found in Arnicae flos are: sesquiterpene lactones, flavonoids, phenolic acids. The anti-inflammatory effect that justifies most of the uses of arnica is primarily due to helenalin derivatives, which are currently being studied, among others, in terms of anticancer activity and against selected parasitic protozoa causing tropical diseases. The development of new solutions for obtaining large amounts of arnica material for the pharmaceutical and cosmetics industries, as well as the search for an efficient source of helenalin derivatives, is therefore justified by biological activity and the expanding range of possible medicinal applications. The high demand for plant raw materials and the prohibited collection from natural sites of strictly protected mountain arnica contributed to the optimization of the cultivation of this species, the acquisition of plant material with comparable qualitative and quantitative characteristics from other species (primarily meadow arnica), as well as the use of biotechnological methods in plant in vitro cultures. In this area, the most numerous publications concern the development of effective micro-propagation of arnica for cultivation. Biotechnological procedures used to develop methods for obtaining helenalin derivatives from plant in vitro cultures have value as the natural science research as well as economic importance and this potential may also be covered by patent protection. In this study, we present the traditional uses of mountain arnica in Europe in the 20th century and summarize the current directions of research in the 21st century, with particular emphasis on the use of plant in vitro cultures. Besides historical documents, the cited works are related to obtaining valuable plant material from arnica based on selected review publications and original research works available in publication databases (PubMed, ScienceDirect). There are also mentioned plants of other species belonging to Asteraceae family, commonly called „arnica” in the regions of Brazil or Mexico (known as: Brasilian arnica, Mexican arnica, false arnica), which are the subject of research works on detailed phytochemical analysis and biological activity, including their anti-inflammatory effect.

The red chili is a superior horticultural product and are currently still a commodity contributing to significant inflation in the Jambi area. The red chili harvest in Jambi Province in 2020 was 10.77 tons ha-1, with national productivity of 12 tons ha-1, while the potential for chili production could exceed 20 tons ha-1. The cause of low chili productivity in Indonesia is the limitation of obtaining superior chili varieties that have stable superior characteristics in different agro-climates, and are less resistant to environmental stress. Environmental stress is the accumulation of the impact of farmers' dependence on intensive and excessive use of inorganic fertilizers and pesticides, which has an impact on high production costs, the phenomenon of pest and other pathogen resurgence, the death of natural enemies, environmental pollution and threats to the safety of the products produced. The research was carried out to overcome the problem of low productivity of chili plants, especially in Jambi Province, consistency in the Application and Development of Environmentally Friendly Technology. The research results showed that the application of Tricho-tithonia 20 tons ha-1 and the recommended dose of 50% NPK fertilizer showed the best growth response and results in local-Jambi red chili varieties. The local varieties tested also have good adaptability to agro-climates that are different from the areas of origin of the chili varieties.

ABSTRACTGene technologies, such as transgenesis and new breeding techniques (NBTs), expand the toolbox for plant breeding. Many countries in Africa, however, have long been seen as “slow adopters” of gene technologies for several reasons, one being the lack of, or overly restrictive, biosafety regulatory frameworks. This is sometimes attributed to the influence of the precautionary-oriented EU biosafety policies. This study analyses and compares the biosafety regulatory frameworks and their implementation in Kenya, Nigeria and Uganda, and in the EU member state Sweden. The focus is on (1) the structure of the biosafety regulatory frameworks including the scope of the legislation, (2) the duration and cost of regulatory authorization for field trials with genetically modified (GM) plants, and (3) the regulatory approach to NBT products, i.e. to what extent NBT products are subject to the provisions of the biosafety regulatory framework. The data was collected through studying relevant legal and policy documents as well as interviewing regulatory officers and researchers in the respective countries. We found that the regulatory procedures in the selected countries are relatively straightforward, while the costs and duration may present a challenge. The regulatory approach to NBT products differ between the selected African countries and Sweden, the latter which follows EU regulations. The results are discussed in terms of the impact the regulatory developments in these four jurisdictions may have on international R&D collaborations involving the use of gene technologies and we also weigh the results against the common conception that Europe exerts a heavy influence on African countries in this technology field.

Grape is one of the most economically significant berry crops. Owing to the biological characteristics of grapes, such as the long juvenile period (5–8 years), high degree of genome heterozygosity, and the frequent occurrence of inbreeding depression, homozygosity during crossbreeding leads to loss of varietal characteristics and viability. CRISPR/Cas editing has become the tool of choice for improving elite technical grape varieties. This study provides the first evidence of a decrease in the total fraction of phenolic compounds and an increase in the concentration of peroxide compounds in grape callus cells upon the addition of chitosan to the culture medium. These previously unreported metabolic features of the grape response to chitosan have been described and used for the first time to increase the probability of selecting plant cells with MLO7 knockout characterised by an oxidative burst in response to the presence of a pathogen modulated by chitosan in the high-metabolite black grape variety ‘Merlot’. This was achieved by using a CRISPR/Cas9 editing vector construction with the peroxide sensor HyPer as a reporter. This research represents the first CRISPR/Cas9 editing of ‘Merlot’, one of the most economically important elite technical grape varieties.

Key message: Characterization of Physcomitrella 3′UTRs across different promoters yields endogenous single and double terminators for usage in molecular pharming. The production of recombinant proteins for health applications accounts for a large share of the biopharmaceutical market. While many drugs are produced in microbial and mammalian systems, plants gain more attention as expression hosts to produce eukaryotic proteins. In particular, the good manufacturing practice (GMP)-compliant moss Physcomitrella (Physcomitrium patens) has outstanding features, such as excellent genetic amenability, reproducible bioreactor cultivation, and humanized protein glycosylation patterns. In this study, we selected and characterized novel terminators for their effects on heterologous gene expression. The Physcomitrella genome contains 53,346 unique 3′UTRs (untranslated regions) of which 7964 transcripts contain at least one intron. Over 91% of 3′UTRs exhibit more than one polyadenylation site, indicating the prevalence of alternative polyadenylation in Physcomitrella. Out of all 3′UTRs, 14 terminator candidates were selected and characterized via transient Dual-Luciferase assays, yielding a collection of endogenous terminators performing equally high as established heterologous terminators CaMV35S, AtHSP90, and NOS. High performing candidates were selected for testing as double terminators which impact reporter levels, dependent on terminator identity and positioning. Testing of 3′UTRs among the different promoters NOS, CaMV35S, and PpActin5 showed an increase of more than 1000-fold between promoters PpActin5 and NOS, whereas terminators increased reporter levels by less than tenfold, demonstrating the stronger effect promoters play as compared to terminators. Among selected terminator attributes, the number of polyadenylation sites as well as polyadenylation signals were found to influence terminator performance the most. Our results improve the biotechnology platform Physcomitrella and further our understanding of how terminators influence gene expression in plants in general. [ABSTRACT FROM AUTHOR]

AbstractPlants, anchored throughout their life cycles, face a unique set of challenges from fluctuating environments and pathogenic assaults. Central to their adaptative mechanisms are transcription factors (TFs), particularly the AP2/ERF superfamily—one of the most extensive TF families unique to plants. This family plays instrumental roles in orchestrating diverse biological processes ranging from growth and development to secondary metabolism, and notably, responses to both biotic and abiotic stresses. Distinguished by the presence of the signature AP2 domain or its responsiveness to ethylene signals, the AP2/ERF superfamily has become a nexus of research focus, with increasing literature elucidating its multifaceted roles. This review provides a synoptic overview of the latest research advancements on the AP2/ERF family, spanning its taxonomy, structural nuances, prevalence in higher plants, transcriptional and post-transcriptional dynamics, and the intricate interplay in DNA-binding and target gene regulation. Special attention is accorded to the ethylene response factor B3 subgroup protein Pti5 and its role in stress response, with speculative insights into its functionalities and interaction matrix in tomatoes. The overarching goal is to pave the way for harnessing these TFs in the realms of plant genetic enhancement and novel germplasm development. [ABSTRACT FROM AUTHOR]