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Frontiers in Plant Science 2023(L.) Gaertn. is an exceptional crop known for its remarkable health benefits, high levels of beneficial polyphenols and gluten-free properties, making it highly...
(L.) Gaertn. is an exceptional crop known for its remarkable health benefits, high levels of beneficial polyphenols and gluten-free properties, making it highly sought-after as a functional food. Its self-fertilisation capability and adaptability to challenging environments further contribute to its potential as a sustainable agricultural option. To harness its unique traits, genetic transformation in is crucial. In this study, we optimised the Agrobacterium-mediated transformation protocol for callus, resulting in a transformation rate of regenerated plants of approximately 20%. The protocol's effectiveness was confirmed through successful GUS staining, GFP expression, and the generation of albino plants via gene inactivation. These results validate the feasibility of genetic manipulation and highlight the potential for trait enhancement in .
PubMed: 37746024
DOI: 10.3389/fpls.2023.1270150 -
Current Opinion in Microbiology Jun 2024The governing principles and suites of genes for lateral elongation or incorporation of new cell wall material along the length of a rod-shaped cell are well described.... (Review)
Review
The governing principles and suites of genes for lateral elongation or incorporation of new cell wall material along the length of a rod-shaped cell are well described. In contrast, relatively little is known about unipolar elongation or incorporation of peptidoglycan at one end of the rod. Recent work in three related model systems of unipolar growth (Agrobacterium tumefaciens, Brucella abortus, and Sinorhizobium meliloti) has clearly established that unipolar growth in the Hyphomicrobiales order relies on a set of genes distinct from the canonical elongasome. Polar incorporation of envelope components relies on homologous proteins shared by the Hyphomicrobiales, reviewed here. Ongoing and future work will reveal how unipolar growth is integrated into the alphaproteobacterial cell cycle and coordinated with other processes such as chromosome segregation and cell division.
Topics: Brucella abortus; Sinorhizobium meliloti; Bacterial Proteins; Agrobacterium tumefaciens; Cell Wall; Peptidoglycan; Cell Division
PubMed: 38569420
DOI: 10.1016/j.mib.2024.102470 -
Microbiology Resource Announcements Nov 2023We report the complete genome of ARqua1 generated from Oxford Nanopore and Illumina sequencing. The genome of ARqua1 has a total length of 5,714,310 bp, comprising a...
We report the complete genome of ARqua1 generated from Oxford Nanopore and Illumina sequencing. The genome of ARqua1 has a total length of 5,714,310 bp, comprising a circular chromosome, a linear chromosome, and two plasmids. In total, 5,446 genes were predicted, of which 5,288 were annotated.
PubMed: 37812076
DOI: 10.1128/MRA.00554-23 -
Journal of Bacteriology Jul 2023The transcriptional regulator PecS is encoded by select bacterial pathogens. For instance, in the plant pathogen Dickeya dadantii, PecS controls a range of virulence...
The transcriptional regulator PecS is encoded by select bacterial pathogens. For instance, in the plant pathogen Dickeya dadantii, PecS controls a range of virulence genes, including pectinase genes and the divergently oriented gene , which encodes an efflux pump through which the antioxidant indigoidine is exported. In the plant pathogen (formerly named Agrobacterium tumefaciens), the locus is conserved. Using a strain of in which has been disrupted, we show here that PecS controls a range of phenotypes that are associated with bacterial fitness. PecS represses flagellar motility and chemotaxis, which are processes that are important for to reach plant wound sites. Biofilm formation and microaerobic survival are reduced in the disruption strain, whereas the production of acyl homoserine lactone (AHL) and resistance to reactive oxygen species (ROS) are increased when is disrupted. AHL production and resistance to ROS are expected to be particularly relevant in the host environment. We also show that PecS does not participate in the induction of genes. The inducing ligands for PecS, urate, and xanthine, may be found in the rhizosphere, and they accumulate within the plant host upon infection. Therefore, our data suggest that PecS mediates fitness during its transition from the rhizosphere to the host plant. PecS is a transcription factor that is conserved in several pathogenic bacteria, where it regulates virulence genes. The plant pathogen is important not only for its induction of crown galls in susceptible plants but also for its role as a tool in the genetic manipulation of host plants. We show here that PecS controls a range of phenotypes, which would confer the bacteria an advantage while transitioning from the rhizosphere to the host plant. This includes the production of signaling molecules, which are critical for the propagation of the tumor-inducing plasmid. A more complete understanding of the infection process may inform approaches by which to treat infections as well as to facilitate the transformation of recalcitrant plant species.
Topics: Transcription Factors; Reactive Oxygen Species; Gene Expression Regulation, Bacterial; Agrobacterium; Agrobacterium tumefaciens; Bacterial Proteins
PubMed: 37314346
DOI: 10.1128/jb.00478-22 -
Methods in Molecular Biology (Clifton,... 2024Bimolecular fluorescence complementation (BiFC) is an assay widely used for studying protein-protein interactions and determining the subcellular localization of...
Bimolecular fluorescence complementation (BiFC) is an assay widely used for studying protein-protein interactions and determining the subcellular localization of proteins. This technique involves fusing the proteins of interest to separate structural domains of a fluorescent protein, followed by transient expression in cells. The interaction between the proteins of interest in vivo allows the reconstitution of the fluorescence that can be visualized by fluorescence microscopy. BiFC has been particularly useful in investigating the interactions between viral and host proteins. Here, we describe the steps involved in preparing expression cassettes that allow the expression of proteins of interest fused to nonfluorescent fragments of yellow fluorescent protein (YFP), Agrobacterium transformations, and agroinfiltration of Nicotiana benthamiana leaves to facilitate virus protein-host protein interactions. Finally, high-resolution images can be obtained by analyzing the leaves under a confocal microscope.
Topics: Host Microbial Interactions; Microscopy, Fluorescence; Agrobacterium; Biological Assay; Coloring Agents
PubMed: 37987908
DOI: 10.1007/978-1-0716-3485-1_15 -
Plant Disease Jan 2024Pathogenic and are phytobacteria that induce crown gall and leafy gall disease, respectively, resulting in undesirable growth abnormalities. When present in nurseries,...
Pathogenic and are phytobacteria that induce crown gall and leafy gall disease, respectively, resulting in undesirable growth abnormalities. When present in nurseries, plants infected by either bacterium are destroyed, resulting in substantial losses for growers, especially those producing plants valued for their ornamental attributes. There are many unanswered questions regarding pathogen transmission on tools used to take cuttings for propagation and whether products used for bacterial disease control are effective. We investigated the ability to transmit pathogenic and on secateurs and the efficacy of registered control products against both bacteria in vitro and in vivo Experimental plants used were × , × , and × for and × and 'Siskiyou' with . In separate experiments, we found secateurs could convey both bacteria in numbers sufficient to initiate disease in a host-dependent manner and that bacteria could be recovered from secateurs after a single cut through an infected stem. In in vivo assays, none of six products tested against prevented crown gall disease, although several products appeared promising in in vitro trials. Likewise, four compounds trialed against . failed to prevent disease. Sanitation and clean planting material remain the primary means of disease management.
Topics: Agrobacterium tumefaciens; Plant Tumors; Rhodococcus; Plants
PubMed: 37368442
DOI: 10.1094/PDIS-11-22-2557-RE -
Applied Microbiology and Biotechnology Oct 2023Woodfordia fruticosa (L.) Kurz is a woody medicinal shrub (Lythraceae) commonly known as the "fire flame bush." W. fruticosa plant parts either alone or whole plant have... (Review)
Review
Woodfordia fruticosa (L.) Kurz is a woody medicinal shrub (Lythraceae) commonly known as the "fire flame bush." W. fruticosa plant parts either alone or whole plant have a long history of recommended use in the Indian medicine systems of Ayurveda, Unani, and Siddha (AUS). This plant is prominently known for its pharmacological properties, viz., antimicrobial, anti-inflammatory, anti-peptic ulcer, hepatoprotective, immunomodulatory, antitumor, cardioprotective, analgesic, and wound healing activities. Its important phyto-constituents, woodfordin C, woodfordin I, oenothein B, and isoschimacoalin-A, exhibit in vitro or in vivo physiological activities beneficial to human health. As the plant is a rich storehouse of phyto-constituents, it is indiscriminately used in its wild habitats. Moreover, due to very poor seed viability and difficult-to-root qualities, it is placed under IUCN list of endangered plant species. For W. fruticosa, biomass production or to its conservation by in vitro regeneration is the best feasible alternative. Till date, only few important in vitro regeneration methods are reported in W. fruticosa. ISSR molecular markers based clonal fidelity and Agrobacterium-mediated transformation has been demonstrated, indicating that W. fruticosa is amenable to genetic manipulation and genome editing studies. This review presents concise summary of updated reports on W. fruticosa phyto-constituents and their biological activities, while a critical appraisal of biotechnological interventions, shortcomings, and factors influencing such potential areas success was presented. The unexplored gaps addressed here are relevant for W. fruticosa scientific innovations yet to come. In this paper, for the first time, we have presented a simple and reproducible protocol for synthetic seed production in W. fruticosa. KEY POINTS: • Critical and updated records on W. fruticosa phytochemistry and its activities • In vitro propagation and elicitation of secondary metabolites in W. fruticosa • Key bottlenecks, in vitro flowering, value addition, and outlook in W. fruticosa.
Topics: Humans; Plant Extracts; Woodfordia; Anti-Infective Agents; Anti-Inflammatory Agents; Wound Healing
PubMed: 37522947
DOI: 10.1007/s00253-023-12695-x -
Frontiers in Plant Science 2023Woody plants play a vital role in global ecosystems and serve as valuable resources for various industries and human needs. While many woody plant genomes have been... (Review)
Review
Woody plants play a vital role in global ecosystems and serve as valuable resources for various industries and human needs. While many woody plant genomes have been fully sequenced, gene function research and biotechnological breeding advances have lagged behind. As a result, only a limited number of genes have been elucidated, making it difficult to use newer tools such as CRISPR-Cas9 for biotechnological breeding purposes. The use of as a transformative tool in plant biotechnology has received considerable attention in recent years, particularly in the research field on woody plants. Over the past three decades, numerous woody plants have been effectively transformed using -mediated techniques. Some of these transformed plants have successfully regenerated. Recent research on -mediated transformation of woody plants has demonstrated its potential for various applications, including gene function analysis, gene expression profiling, gene interaction studies, and gene regulation analysis. The introduction of the Ri plasmid has resulted in the emergence of several Ri phenotypes, such as compact plant types, which can be exploited for Ri breeding purposes. This review paper presents recent advances in A. rhizogenes-mediated basic research and Ri breeding in woody plants. This study highlights various aspects of A. rhizogenes-mediated transformation, its multiple applications in gene function analysis, and the potential of Ri lines as valuable breeding materials.
PubMed: 38034586
DOI: 10.3389/fpls.2023.1196561 -
Animals : An Open Access Journal From... Jul 2023The objectives of this review are to investigate the quantitative, compositional, and structural differences of β-glucans and the functional effects of β-glucans on... (Review)
Review
The objectives of this review are to investigate the quantitative, compositional, and structural differences of β-glucans and the functional effects of β-glucans on the intestinal health and growth of nursery pigs. Banning antibiotic feed supplementation increased the research demand for antibiotic alternatives to maintain the intestinal health and growth of nursery pigs. It has been proposed that β-glucans improve the growth efficiency of nursery pigs through positive impacts on their intestinal health. However, based on their structure and source, their impacts can be extensively different. β-glucans are non-starch polysaccharides found in the cell walls of yeast (), bacteria, fungi (), and cereal grains (mainly barley and oats). The total β-glucan content from cereal grains is much greater than that of microbial β-glucans. Cereal β-glucans may interfere with the positive effects of microbial β-glucans on the intestinal health of nursery pigs. Due to their structural differences, cereal β-glucans also cause digesta viscosity, decreasing feed digestion, and decreasing nutrient absorption in the GIT of nursery pigs. Specifically, cereal β-glucans are based on linear glucose molecules linked by β-(1,3)- and β-(1,4)-glycosidic bonds with relatively high water-soluble properties, whereas microbial β-glucans are largely linked with β-(1,3)- and β-(1,6)-glycosidic bonds possessing insoluble properties. From the meta-analysis, the weight gain and feed intake of nursery pigs increased by 7.6% and 5.3%, respectively, through the use of yeast β-glucans (from ), and increased by 11.6% and 6.9%, respectively, through the use of bacterial β-glucans (from sp.), whereas the use of cereal β-glucans did not show consistent responses. The optimal use of yeast β-glucans () was 50 mg/kg in nursery pig diets based on a meta-analysis. Collectively, use of microbial β-glucans can improve the intestinal health of nursery pigs, enhancing immune conditions, whereas the benefits of cereal β-glucans on intestinal health were not consistent.
PubMed: 37444033
DOI: 10.3390/ani13132236 -
Journal of Dental Sciences Oct 2023Oral flora is related to various immune-related diseases. Herein we explored the characteristics of oral flora in patients with pemphigus vulgaris (PV) and analyzed the...
BACKGROUND/PURPOSE
Oral flora is related to various immune-related diseases. Herein we explored the characteristics of oral flora in patients with pemphigus vulgaris (PV) and analyzed the correlation between oral flora and PV.
MATERIALS AND METHODS
Twenty-two untreated patients with PV and 12 healthy controls (HC) were included in this case-control study. The characteristics of salivary microbiome were assessed by high-throughput sequencing using the 16S rRNA Illumina MiSeq approach, and differences between the PV and HC groups were determined. The Kyoto Encyclopedia of Genes and Genomes (KEGG) database was applied to screen key metabolic pathways and preliminarily explore potential mechanisms underlying PV occurrence and development.
RESULTS
The abundance of oral flora in the PV group was significantly lower than that in the HC group, and there were characteristic changes. The relative abundance of and in the PV group was significantly higher than that in the HC group ( < 0.05) and that of , , and was significantly lower ( < 0.05). There was a linear correlation between and serum Dsg3 level in PV. KEGG pathway analyses indicated significant differences in nine metabolic pathways between the PV and HC groups ( < 0.05), namely carbohydrate metabolism, digestive system, neurodegenerative disease, glycan biosynthesis and metabolism, drug resistance: antimicrobial, infectious disease: viral, circulatory system, excretory system, and nervous system.
CONCLUSION
The oral flora of patients with PV presented characteristic changes, and several metabolic pathways were affected, including N-glycan biosynthesis and metabolism. spp. appear to require the most attention in PV. We believe that oral flora dysbacteriosis contributes to PV occurrence and development.
PubMed: 37799932
DOI: 10.1016/j.jds.2023.01.037