Stem Tissue Research Articles

The Impact of Bone Marrow Venting Augmentation on Knee Functions in the Repair of Vertical/Longitudinal Meniscus Tears: A Triple Comparison.

Preserving and restoring meniscus function is crucial for maintaining knee biomechanics and overall functionality. To enhance the healing process after meniscus repair, various biological techniques have been introduced. In this study, three treatment approaches examined were bone marrow venting with stem cells (BMVP), anterior cruciate ligament reconstruction (ACLR) with meniscus repair, and isolated meniscus repair. From 2015 to 2019, we retrospectively analyzed data from 83 patients who presented with complaints of knee pain or pain in addition to instability, were diagnosed with vertical/longitudinal meniscus tears, and underwent arthroscopic repair (30 with isolated repair, 28 with ACLR repair, and 25 with BMVP) at our University Hospital. Among the 28 patients with anterior cruciate ligament rupture, pain and instability were the predominant complaints, whereas 55 patients with meniscus tear primarily complained pain. Clinical and functional conditions were assessed using Lysholm, WOMAC, IKDC, and VAS scores before and 18months after surgery. Surgical success was evaluated based on Barrett's criteria. Causes of failure were analyzed considering demographic data, smoking status, injury time, tear location, shape, zone, and suture type and number. Of the participants, 58 were male, 25 were female, 61 had medial meniscus lesions, and 22 had lateral meniscus lesions. The tear types included 51 simple longitudinal tears, 18 bucket handle tears, and 14 complex tears. A significant improvement was observed in the postoperative 18month Lysholm, WOMAC, and IKDC values in all three groups (p: 0.001). At the 18month mark, clinical and functional outcomes were comparable between BMVP and ACLR repair groups (Lysholm p: 0.951, WOMAC p: 0.241, IKDC p: 0.984). Both of these procedures yielded better results compared to isolated meniscus repair (Lysholm p: 0.001, WOMAC p: 0.027, IKDC p: 0.001). The superior clinical and functional outcomes observed after meniscus repair with BMVP and ACLR, compared to isolated meniscus repair, indicate positive effects of bone marrow stem cells, blood components, and tissue healing factors on meniscus repair and knee function.

Anatomical limitations in adventitious root formation revealed by magnetic resonance imaging, infrared spectroscopy, and histology of rose genotypes with contrasting rooting phenotypes.

Adventitious root (AR) formation is one of the most important developmental processes in vegetative propagation. Although genotypic differences in rose rooting ability are well known, the causal factors are not well understood. The rooting of two contrasting genotypes, 'Herzogin Friederike' and 'Mariatheresia', was compared following a multiscale approach. Using magnetic resonance imaging, we non-invasively monitored the inner structure of stem cuttings during initiation and progression of AR formation for the first time. Spatially resolved Fourier-transform infrared spectroscopy characterized the chemical composition of the tissues involved in AR formation. The results were validated through light microscopy and complemented by immunolabelling. The outcome demonstrated similarity of both genotypes in root primordia formation, which did not result in root protrusion through the shoot cortex in the difficult-to-root genotype 'Mariatheresia'. The biochemical composition of the contrasting genotypes highlighted main differences in cell wall-associated components. Further spectroscopic analysis of 15 contrasting rose genotypes confirmed the biochemical differences between easy- and difficult-to-root groups. Collectively, our data indicate that it is not the lack of root primordia limiting AR formation in these rose genotypes, but the firmness of the outer stem tissue and/or cell wall modifications that pose a mechanical barrier and prevent root extension and protrusion.

Integrating Targeted Metabolomics and Targeted Proteomics to Study the Responses of Wheat Plants to Engineered Nanomaterials.

This manuscript presents a multiomics investigation into the metabolic and proteomic responses of wheat to molybdenum (Mo)- and copper (Cu)-based engineered nanomaterials (ENMs) exposure via root and leaf application methods. Wheat plants underwent a four-week growth period with a 16 h photoperiod (light intensity set at 150 μmol·m-2·s-1), at 22 °C and 60% humidity. Six distinct treatments were applied, including control conditions alongside exposure to Mo- and Cu-based ENMs through both root and leaf routes. The exposure dosage amounted to 6.25 mg of the respective element per plant. An additional treatment with a lower dose (0.6 mg Mo/plant) of Mo ENM exclusively through the root system was introduced upon the detection of phytotoxicity. Utilizing LC-MS/MS analysis, 82 metabolites across various classes and 24 proteins were assessed in different plant tissues (roots, stems, leaves) under diverse treatments. The investigation identified 58 responsive metabolites and 19 responsive proteins for Cu treatments, 71 responsive metabolites, and 24 responsive proteins for Mo treatments, mostly through leaf exposure for Cu and root exposure for Mo. Distinct tissue-specific preferences for metabolite accumulation were revealed, highlighting the prevalence of organic acids and fatty acids in stem or root tissues, while sugars and amino acids were abundant in leaves, mirroring their roles in energy storage and photosynthesis. Joint-pathway analysis was conducted and unveiled 23 perturbed pathways across treatments. Among these, Mo exposure via roots impacted all identified pathways, whereas exposure via leaf affected 15 pathways, underscoring the reliance on exposure route of metabolic and proteomic responses. The coordinated response observed in protein and metabolite concentrations, particularly in amino acids, highlighted a dynamic and interconnected proteomic-to-metabolic-to-proteomic relationship. Furthermore, the contrasting expression patterns observed in glutamate dehydrogenase (upregulation at 1.38 ≤ FC ≤ 1.63 with high Mo dose, and downregulation at 0.13 ≤ FC ≤ 0.54 with low Mo dose) and its consequential impact on glutamine expression (7.67 ≤ FC ≤ 39.60 with high Mo dose and 1.50 ≤ FC ≤ 1.95 with low Mo dose) following Mo root exposure highlighted dose-dependent regulatory trends influencing proteins and metabolites. These findings offer a multidimensional understanding of plant responses to ENMs exposure, guiding agricultural practices and environmental safety protocols while advancing knowledge on nanomaterial impacts on plant biology.

Liner Propagation Method Influences Growth of Container Grown Bottlebrush

Bottlebrush (Callistemon vinimalis) is a widely propagated and cultivated ornamental large shrub with large red bottlebrush-like flowers. Traditional clonal propagation using stem cuttings may be replaced with tissue cultured liners. In this study, we established a container-grown field experiment of bottlebrush ‘Little John’ using liners propagated from both rooted stem cuttings and tissue culture. Growth index was recorded by propagation method periodically through the 34-week period, and both fresh and dry weights of roots and shoots recorded at experiment’s end. Final growth index of plants grown from tissue cultured liners were significantly greater than growth index of plants started from rooted stem cuttings. Both fresh and dry root weight means were significantly greater in plants propagated by tissue culture. Further testing of containerized bottlebrush production, through the flowering stage, will better determine whether tissue-cultured liners accelerate production time vs. liners from stem cuttings.

Evaluation of volatile components from the tuber, fibrous roots, bud, stem and leaf tissues of Bletilla striata for its anti-colon cancer activity.

Bletilla striata (Thunb.) Rchb.f., a medicinal plant in the Orchidaceae family, is mainly found in East Asia and has extensive pharmacological activities. Plant's volatile components are important active ingredients with a wide range of physiological activities, and B. striata has a special odor and unique volatile components. Yet it has received little attention, hindering a full understanding of its phytochemical components. Employing the ultrasonic-assisted extraction method, the volatile components of B. striata's fibrous root, bud, aerial part and tuber were extracted, resulting in yields of 0.06%, 0.64%, 3.38% and 4.47%, respectively. A total of 78 compounds were identified from their chemical profiles using gas chromatography-mass spectrometry (GC-MS), including 45 components with the main compounds of linoleic acid (content accounting for 31.23%), n-hexadecanoic acid (13.53%), and octadecanoic acid (9.5%) from the tuber, 34 components with the main compounds of eicosane, 2-methyl- (28.42%), linoelaidic acid (10.43%), linoleic acid (4.53%), and n-hexadecanoic acid (6.91%) from the fibrous root, 38 components with the main compounds of pentadeca-6,9-dien-1-ol (9.29%), n-hexadecanoic acid (11%), eicosane,2-methyl- (23.43%), and linoleic acid (23.53%) from the bud, and 27 components with the main compounds of linoelaidic acid (5.97%), n-hexadecanoic acid (15.99%), and linolenic acid ethyl ester (18.9%) from the aerial part. Additionally, the growth inhibition activity against colon cancer HCT116 cells was evaluated using sulforhodamine B (SRB) assay and the thiazolyl blue tetrazolium bromide (MTT) assay, and the accumulation of reactive oxygen species (ROS) was determined using dichloro-dihydro-fluorescein diacetate (DCFH-DA) staining and fluorescence intensity analysis. The volatile extracts exhibited significant growth inhibitory efficacy against HCT116 cells, with half-maximal inhibitory concentration (IC50) values of 3.65, 2.32, 2.42 and 3.89mg/mL in the SRB assay, and 3.55, 2.58, 3.12 and 4.80mg/mL in the MTT assay for the root, bud, aerial part, and tuber, respectively. Notably, treatment with the aerial part extract caused morphological changes in the cells and significantly raised the intracellular ROS level. In summary, the chemical profiles of the volatile components of B. striata were revealed for the first time, demonstrating a certain tissue specificity. Additionally, it demonstrated for the first time that these volatile extracts possess potent anti-colon cancer activity, highlighting the importance of these volatile components in B. striata's medicinal properties.

Oriental beech decline caused by Brenneria alni

AbstractHyrcanian forests comprise a long strip along the southern coast of the Caspian Sea in northern Iran. Oriental beech (Fagus orientalis) is one of the most valuable forest tree species in the Hyrcanian forests. Bark necrosis and bleeding spots of oriental beech were observed in Savadkuh forests in Mazandaran province during summer 2021. Ten samples were collected from symptomatic oriental beech trees. The bark canker symptoms included stem tissue necrosis, dark brown to black spots on the trunk, bleeding with dark exudates and dark lesions in the inner bark. Twenty‐six strains of culturable bacteria were isolated from the samples which in culture were predominantly round, convex, smooth with entire margins on nutrient agar and a metallic green sheen pigment on eosin‐methylene blue (EMB) agar. Fourteen strains induced necrosis and browning of the tissues around the inoculation sites on acorns after 2 weeks. Five representative strains also caused dieback in twigs of oriental beech 3 weeks after inoculation. These bacteria were Gram‐negative, catalase positive and oxidase negative. All strains produced colonies with a green metallic sheen on EMB agar. Strains were negative in tests for starch and gelatine hydrolysis or production of levan. A phylogenetic tree based on concatenated sequences of gyrB and infB genes, of representative strains well as in each of the single gene‐based phylogenetic trees, indicated that these strains clustered with Brenneria alni NCPPB 3934T with high bootstrap values. To the best of our knowledge, this is the first report of B. alni associated with bark necrosis and bleeding spots on oriental beech.

Modelling metabolic fluxes of tomato stems reveals that nitrogen shapes central metabolism for defence against Botrytis cinerea.

Among plant pathogens, the necrotrophic fungus Botrytis cinerea is one of the most prevalent, leading to severe crop damage. Studies related to its colonization of different plant species have reported variable host metabolic responses to infection. In tomato, high N availability leads to decreased susceptibility. Metabolic flux analysis can be used as an integrated method to better understand which metabolic adaptations lead to effective host defence and resistance. Here, we investigated the metabolic response of tomato infected by B. cinerea in symptomless stem tissues proximal to the lesions for 7 d post-inoculation, using a reconstructed metabolic model constrained by a large and consistent metabolic dataset acquired under four different N supplies. An overall comparison of 48 flux solution vectors of Botrytis- and mock-inoculated plants showed that fluxes were higher in Botrytis-inoculated plants, and the difference increased with a reduction in available N, accompanying an unexpected increase in radial growth. Despite higher fluxes, such as those involved in cell wall synthesis and other pathways, fluxes related to glycolysis, the tricarboxylic acid cycle, and amino acid and protein synthesis were limited under very low N, which might explain the enhanced susceptibility. Limiting starch synthesis and enhancing fluxes towards redox and specialized metabolism also contributed to defence independent of N supply.

First Report of Phytophthora ramorum Causing Leaf Spots on Cornus capitata (Evergreen Dogwood) in United States.

Cornus capitata Wall. ex Roxb. (evergreen dogwood) is a bushy evergreen tree or shrub native to East Asia grown for its showy creamy bracts in late spring followed by attractive red fruit. In Feb 2023, a sample of foliage with leaf spots and tip dieback from C. capitata 'Mountain Moon' was submitted from a Humboldt Co. nursery as part of a CDFA inspection program for Phytophthora ramorum. The leaf spots were medium to dark brown, irregularly shaped, and ranged from 5 to 8 mm in diameter. They were located primarily along the leaf midrib and covered up to 1/4 of the leaf surface. Six 6-mm-diameter leaf discs taken from the margins of brown lesions and tip dieback were plated on Phytophthora selective CMA-PARP (PARP) media (Jeffers and Martin 1986). After 6 to 10 days, colonies resembling P. ramorum, with coralloid coenocytic hyphae, chlamydospores, ellipsoidal semi-papillate and caducous sporangia, and a relatively slow growth rate were recovered. Abundant sporangia formed on agar singly or in clusters on sympodially branched sporangiophores (n = 50), varying in size from 35 to 60 μm × 20 to 30 μm (mean 45.6 × 24.8 μm) with a length/breadth ratio ranging from 1.3 to 2.3 (mean 1.8). Chlamydospores (n = 50) ranged from 35 to 62 μm in diameter (mean 51.9 μm) on 14-day-old PARP cultures. The internal transcribed spacer region (ITS) using primers ITS5/ITS4 (White et al. 1990; accession no. OR636225) and cytochrome oxidase subunit 1 region (cox1) using primers OomCox1Levup/Fm85mod (Robideau et al. 2011; OR635665) of one isolate (0254-32A) were amplified and sequenced. BLAST analysis showed 100% identity of both regions to P. ramorum ex-type strain (MG865581 and MH136973). Microsatellite loci placed the P. ramorum isolate in the NA2 clonal lineage (Goss et al, 2011). Pathogenicity of P. ramorum isolate 0254-32A was tested using five C. capitata plants (2.5-year-old, 28-cm-tall, 3.78-liter pot). Zoospore inoculum was produced as described in Blomquist et al. (2021). Above ground parts of each plant were sprayed with inoculum (15 ml, 1.3 × 105 zoospores/ml). Inoculated plants were incubated in a dew chamber in the dark at 23°C for 72 h and then placed in a 23±1°C growth chamber with a 12-h photoperiod. Five control plants were treated as above but with sterile water instead of the zoospore suspension. Two days after inoculation, brown spots were visible on leaves on all inoculated plants, initiating from where the drops of inoculum had persisted. After 3 days, brown lesions, from water drop- to majority of entire leaf-sized, were observed on approximately 75% of inoculated leaves. After 6 days, lesions expanded to the edges of leaves, causing leaf curling and defoliation. Lesions stopped expanding after 3 weeks, and by 4 weeks, most infected leaves had abscised, with no new infections observed. Phytophthora ramorum was consistently isolated from foliar lesions of inoculated plants on PARP. It was not isolated from leaf or stem tissues of control plants, which remained asymptomatic during the 4-week experiment period. Phytophthora ramorum was detected on C. capitata in the UK in 2015 (DEFRA 2015). To our knowledge, this is the first report of P. ramorum infecting C. capitata in the United States and the completion of Koch's postulates on any Cornus spp. Incidence on C. capitata in the California nursery was low. However, their proximity to other infected foliar hosts suggests Cornus spp. may present a potential risk for the spread of P. ramorum.

First Report of Achromobacter xylosoxidans Causing Bacterial Stem and Leaf Blight on Cyrilla arida in Tennessee and the United States.

Scrub titi (Cyrilla arida), broadleaf semi-evergreen shrub, is endemic to central Florida. However, its smaller stature, lustrous, dark-green leaves and abundance of white racemes in late spring make it a potential candidate for future use in Southeastern U.S. landscapes. Three-years-old container grown C. arida plants maintained in a shade house at the Nursery Research Center, McMinnville, TN exhibited black leaf lesions and brown stem lesions (Fig. 1a) in April 2023. The disease severity was 25% of the shoot area and the disease incidence was 10% out of 60 plants. Symptomatic stem and leaf tissues were surface sterilized with 0.525% NaOCl for 1 min. Bacterial colonies were white-colored, opaque, round with smooth edges on lysogen broth agar medium after 2 days of incubation at 28°C. Bacteria were gram-negative and non-fluorescent on King's B. Esculin, catalase, and oxidase tests were positive but arginine dihydrolase and gelatine hydrolysis were negative. Bacterial identity was confirmed by sequencing of DNA from pure cultures (strains FBG5290 and FBG5294). The 16S ribosomal RNA, RNA polymerase sigma factor (rpoD), enolase (eno), and NADH-quinone oxidoreductase subunit L (nuoL) genes were amplified and sequenced using the primers 8F/1492R (Galkiewicz et al. 2008), rpoDpF/R (Sarkar and Guttman 2004), enoP1/P2 and nuoLP1/P2 (Spilker et al. 2012), respectively. The sequences were deposited in GenBank with acc. nos.: OR689356, OR689357 (16S); OR751366, OR751367 (rpoD); OR792456, OR792457 (eno); and OR792458, OR792459 (nuoL). The closest identified species to our two identical strains was Achromobacter xylosoxidans (CP054571), showing 99.6%, 95.2%, 96.2%, and 95.0% identity with >99% coverage to the above mentioned gene sequences, respectively. Phylogenetic analysis, using concatenated sequences along with the genome sequences of other closely related taxa (Fig. 2), suggest that A. xylosoxidans is presently the identified species, but given the results of the MLST, it may be that this organism will be classified as new species in the future. The pathogenicity of the strains was confirmed on 1-year-old C. arida by inoculating five plants per strain. Stems were inoculated by depositing 15 µl of bacterial suspension (1x108 CFU/mL) into the stem wounded using a scalpel. The inoculation sites were covered with moist cotton and wrapped with Parafilm. Inoculation was also performed on three leaves per plant by using a needleless syringe to infiltrate bacteria into the intercellular spaces (1x108 CFU/mL). Sterile water was used for five control plants. Plants were kept in a greenhouse at 21-23°C, 70% RH, and 16-h photoperiod. All inoculated plants showed brown lesions in stems (Fig. 1b and 1c) and leaves (Fig. 1d) 7-10 days after inoculation, while control plants remained asymptomatic (Fig. 1e and 1f). The bacteria were re-isolated from inoculated plants and confirmed as A. xylosoxidans using morphological and molecular methods. Achromobacter spp. are commonly known as human pathogens, and cross-kingdom pathogenic bacterium in animal (mice) and fungi (Coprinus comatus) (Ye et al. 2018). However, A. xylosoxidans was recently reported as the causal agent of stem rot of Amorphophallus konjac in China (Wei et al. 2023). To our knowledge, this is the first report of A. xylosoxidans causing bacterial stem and leaf blight of C. arida in Tennessee and the U.S. Identification of this novel disease lays the foundation development of effective management strategies.

Analisis Serapan Hara N, P dan K Jaringan Cabai (Capsicum sp.) pada Variasi Tingkat Kemasaman Tanah

This research tested the effect of soil acidity level on the uptake of macro elements in plant tissue. This research is a type of experimental research that examines the effect of soil acidification levels on the absorption of N, P, and K nutrients in chili plant tissues. This research was conducted on the greenhouse experimental land of the Sambas State Polytechnic, Sambas Regency, West Kalimantan. This activity is carried out from March to October 2022. The research design used a descriptive analysize with three treatments, namely pH = 6 (P1), pH = 7 (P2), and pH = 8 (P3) which were repeated as many as 3 tests. Each treatment consisted of 30 plants planted in a planting medium mixed with soil, burnt husks and animal dung. N, P and K uptake is obtained through the analysis of the covering of stem and leaf tissues. The results showed that the difference in the level of acidification of the growing media did not have a effect on the level of absorption of nitrogen and phosphorus, but level of acidification have effect in potassium absorption in chili plants. Nitrogen content generally decreases in plant tissue samples aged 30 to 45 days after planting, while in plants aged 45 to 50 days after planting, there is enerally an increase in the accumulation of nitrogen in plant tissues. Absorption of phosphorus elements in tissues fluctuates over a period of 30 to 60 days after planting. The phosphorus content found in leaf tissue treatment pH 6 and pH 7 increased from the observation period of 30 to 60 days after planting. Absorption of potassium elements in tissues fluctuates over a period of 30 to 60 days after planting. An increase in Potassium content in stem and leaf tissues occurred in pH 6 treatment samples. Key words : chili, nitrogen, phosphor, potassium, uptake

Overexpression of tocopherol biosynthesis genes in guayule (Parthenium argentatum) reduces rubber, resin and argentatins content in stem and leaf tissues

Natural rubber produced in stems of the guayule plant (Parthenium argentatum) is susceptible to post-harvest degradation from microbial or thermo-oxidative processes, especially once stems are chipped. As a result, the time from harvest to extraction must be minimized to recover high quality rubber, especially in warm summer months. Tocopherols are natural antioxidants produced in plants through the shikimate and methyl-erythtiol-4-phosphate (MEP) pathways. We hypothesized that increased in vivo guayule tocopherol content might protect rubber from post-harvest degradation, and/or allow reduced use of chemical antioxidants during the extraction process. With the objective of enhancing tocopherol content in guayule, we overexpressed four Arabidopsis thaliana tocopherol pathway genes in AZ-2 guayule via Agrobacterium-mediated transformation. Tocopherol content was increased in leaf and stem tissues of most transgenic lines, and some improvement in thermo-oxidative stability was observed. Overexpression of the four tocopherol biosynthesis enzymes, however, altered other isoprenoid pathways resulting in reduced rubber, resin and argentatins content in guayule stems. The latter molecules are mainly synthesized from precursors derived from the mevalonate (MVA) pathway. Our results suggest the existence of crosstalk between the MEP and MVA pathways in guayule and the possibility that carbon metabolism through the MEP pathway impacts rubber biosynthesis.

METTL3-dependent m6A modification of PSEN1 mRNA regulates craniofacial development through the Wnt/β-catenin signaling pathway

Craniofacial malformations, often associated with syndromes, are prevalent birth defects. Emerging evidence underscores the importance of m6A modifications in various bioprocesses such as stem cell differentiation, tissue development, and tumorigenesis. Here, in vivo, experiments with zebrafish models revealed that mettl3-knockdown embryos at 144 h postfertilization exhibited aberrant craniofacial features, including altered mouth opening, jaw dimensions, ethmoid plate, tooth formation and hypoactive behavior. Similarly, low METTL3 expression inhibited the proliferation and migration of BMSCs, HEPM cells, and DPSCs. Loss of METTL3 led to reduced mRNA m6A methylation and PSEN1 expression, impacting craniofacial phenotypes. Co-injection of mettl3 or psen1 mRNA rescued the level of Sox10 fusion protein, promoted voluntary movement, and mitigated abnormal craniofacial phenotypes induced by mettl3 knockdown in zebrafish. Mechanistically, YTHDF1 enhanced the mRNA stability of m6A-modified PSEN1, while decreased METTL3-mediated m6A methylation hindered β-catenin binding to PSEN1, suppressing Wnt/β-catenin signaling. Pharmacological activation of the Wnt/β-catenin pathway partially alleviated the phenotypes of mettl3 morphant and reversed the decreases in cell proliferation and migration induced by METTL3 silencing. This study elucidates the pivotal role of METTL3 in craniofacial development via the METTL3/YTHDF1/PSEN1/β-catenin signaling axis.

Causal association between cardiovascular proteins and membranous nephropathy: a bidirectional Mendelian randomization.

Multiple circulating proteins have been reported to participate in human diseases. However, the association between cardiovascular proteins and membranous nephropathy (MN) remained profoundly elusive. A bidirectional Mendelian randomization (MR) analysis was conducted to explore the causal correlation between ninety cardiovascular proteins and MN. Genome-wide association study (GWAS) data of cardiovascular proteins and MN were all from European research. Inverse variance weighted (IVW) was used as the main approach. Moreover, MR-Egger, weighted median, weighted mode, and simple mode were also performed. Cochrane's Q test, MR-Egger, and MR-PRESSO were conducted for sensitivity analysis. According to IVW method, fatty acid-binding protein and thrombomodulin (TM) were identified as risk factors for MN, while a protective role was detected in tissue-type plasminogen activator. Additionally, MN was associated with an elevated level of macrophage colony-stimulating factor 1, stem cell factor, TM, and tissue factor. Reversely, MN was also correlated with a downregulated level of beta-nerve growth factor, Cathepsin D, hepatocyte growth factor, interleukin-6 receptor subunit alpha, macrophage colony-stimulating factor 1, and myeloperoxidase. In the sensitivity analysis, no significant pleiotropy and heterogeneity was detected. This was the first study to reveal the causal association between cardiovascular proteins and MN. These specific cardiovascular proteins could be novel biomarkers for MN, and is helpful for timely identify the risk of other diseases that might result from MN. However, further clinical studies are needed to confirm our results.

Bacterial root cancer of cherry in the conditions of the Western part of Azerbaijan

Bacterial root cancer or goiter of the root system is a plant disease with symptoms of proliferation of plant stem or root tissues, caused by the transfer of Ti-plasmids from the virulent bacterium Agrobacterium tumefaciens Conn. into plant cells. The article presents the results of studies of bacterial root cancer of cherries in the conditions of the Ganja-Kazakh geographical zone during 2021—2023. In these years of research, it was found that the disease is widespread in cherry orchards in the western part of the country and severely damages trees. The causative agent of the disease from the soil through wounds and cracks in different parts of the root system penetrates into its cells and the incubation period of the disease begins. It lasts (depending on temperature) 4—7 weeks. The resulting cancer growths do not lead to the rapid death of cherry trees. However, the pathogen suppresses their overall growth, viability and reduces their resistance to fungal and other infections, which leads to a significant decrease in yield, and after a while the energy of the plants is depleted, they gradually dry out and eventually die. The bacterium Agrobacterium tumefaciens Conn. has been studied in detail in the laboratory over the years of research. We have developed a system of complex agrotechnical methods for combating bacterial root cancer.

Effect of phenolics on soil microbe distribution, plant growth, and gall formation

Phenolic compounds, abundant secondary metabolites in plants, profoundly influence soil ecosystems, plant growth, and interactions with herbivores. In this study, we explore the intricate relationships between phenolics, soil microbes, and gall formation in Ageratina adenophora (A. adenophora), an invasive plant species in China known for its allelopathic traits. Using metabolomic and microbial profiling, significant differences in soil microbial composition and metabolite profiles were observed between bulk and rhizosphere soil samples. Phenolics influenced bacterial communities, with distinct microbial populations enriched in each soil type. Additionally, phenolics impacted soil metabolic processes, with variations observed in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis between different soil treatments. Analysis of phenolic content in plant and soil samples revealed considerable variations, with higher concentrations observed in certain plant tissues and soil types. Bioactive phenols extracted from plant and soil samples were identified using gas chromatography/mass spectrometry (GC–MS), providing insights into the diverse chemical composition of these compounds. Furthermore, the effects of phenolics on plant growth and gall formation were investigated. Phenols exhibited both stimulatory and inhibitory effects on plant growth, with optimal concentrations promoting emergence but higher concentrations hindering growth. Gall formation was influenced by phenolic concentrations, leading to structural alterations in stem tissue and gall morphology. Histochemical analysis revealed starch and lipid accumulation in gall tissues, indicating metabolic changes induced by phenolics. The presence of phenolics disrupted tissue structures and influenced vascular bundle orientation in gall tissues. Overall, our study highlights the multifaceted roles of phenolic compounds in soil ecosystems, plant development, and gall formation, facilitating the utilization of secondary metabolites in agriculture.

Novel electroporation-based genome editing of carnation plant tissues using RNPs targeting the anthocyanidin synthase gene.

A novel electroporation method for genome editing was performed using plant tissue samples by direct RNPs-introduction in carnation. Genome editing is becoming a very useful tool in plant breeding. In this study, a novel electroporation method was performed for genome editing using plant tissue samples. The objective was to create a flower color mutant using the pink-flowered carnation 'Kane Ainou 1-go'. For this purpose, a ribonucleoprotein consisting of guide RNA and clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) was introduced into the stem tissue to induce mutations in the anthocyanidin synthase (ANS) gene, which is involved in anthocyanin biosynthesis. As the ANS of 'Kane Ainou 1-go' has not been previously isolated, we initially isolated the ANS gene from 'Kane Ainou 1-go' for characterization. Southern hybridization analysis confirmed that the ANS gene was present in the genome as a two-allele gene with a pair of homologous sequences (ANS-1 and 2); these sequences were used as the target for genome editing. Genome editing was performed by introducing #2_single-guide RNA into the stem tissue using the ribonucleoprotein. This molecule was used because it exhibited the highest efficiency in an analysis of cleavage activity against the target sequence in vitro. Cleaved amplified polymorphic sequence analysis of genomic DNA extracted from 85 regenerated individuals after genome editing was performed. The results indicated that mutations in the ANS gene may have been introduced into two lines. Cloning of the ANS gene in these two lines confirmed the introduction of a single nucleotide substitution mutation for ANS-1 in both lines, and a single amino acid substitution in one line. We discussed the possibility of color change by the amino acid substitution, and also the future applications of this technology.

Exploring the role of the LkABCG36 transporter in lignin accumulation

Lignin is a complex biopolymer formed through the condensation of three monomeric precursors known as monolignols. However, the mechanism underlying lignin precursor transport remains elusive, with uncertainty over whether it occurs through passive diffusion or an active energized process. ATP-binding cassette 36 (ABCG36) plays important roles in abiotic stress resistance. In this study, we investigated the transport functions of LkABCG36 (Larix kaempferi) for lignin precursors and the potential effects of LkABCG36 overexpression in plants. LkABCG36 enhanced the ability of tobacco (Nicotiana tabacum) bright yellow-2 (BY-2) cells to resist monolignol alcohol stress. Furthermore, LkABCG36 overexpression promoted lignin deposition in tobacco plant stem tissue. To understand the underlying mechanism, we measured the BY-2 cell ability to export lignin monomers and the uptake of monolignol precursors in inside-out (inverted) plasma membrane vesicles. We found that the transport of coniferyl and sinapyl alcohols is an ATP-dependent process. Our data suggest that LkABCG36 contributes to lignin accumulation in tobacco stem tissues through a mechanism involving the active transport of lignin precursors to the cell wall. These findings shed light on the lignin biosynthesis process, with important implications for enhancing lignin deposition in plants, potentially leading to improved stress tolerance and biomass production.

Searching for mechanisms driving root pressure in Zea mays—a transcriptomic approach

While there are many theories and a variety of innovative datasets contributing to our understanding of the mechanism generating root pressure in vascular plants, we are still unable to produce a specific cellular mechanism for any species. To discover these mechanisms, we used RNA-Seq to explore differentially expressed genes in three different tissues between individual Zea mays plants expressing root pressure and those producing none. Working from the perspective that roots cells are utililizing a combination of osmotic exudation and hydraulic pressure mechanisms to generate positively-pressured flow of water into the xylem from the soil, we hypothesized that differential expression analysis would yield candidate genes coding for membrane transporters, ion channels, ATPases, and hormones with clear relevance to root pressure generation. In basal stem and coarse root tissue, we observed these classes of differentially expressed genes and more, including a strong cytoskeletal remodeling response. Fine roots displayed remarkably little differential expression relevant to root pressure, leading us to conclude that they either do not contribute to root pressure generation or are constitutively expressing root pressure mechanisms regardless of soil water content.

Advances in Treatments for Epidermolysis Bullosa (EB): Emphasis on Stem Cell-Based Therapy.

Epidermolysis bullosa (EB) is a rare genetic dermatosis characterized by skin fragility and blister formation. With a wide phenotypic spectrum and potential extracutaneous manifestations, EB poses significant morbidity and mortality risks. Currently classified into four main subtypes based on the level of skin cleavage, EB is caused by genetic mutations affecting proteins crucial for maintaining skin integrity. The management of EB primarily focuses on preventing complications and treating symptoms through wound care, pain management, and other supportive measures. However, recent advancements in the fields of stem cell therapy, tissue engineering, and gene therapy have shown promise as potential treatments for EB. Stem cells capable of differentiating into skin cells, have demonstrated positive outcomes in preclinical and early clinical trials by promoting wound healing and reducing inflammation. Gene therapy, on the other hand, aims to correct the underlying genetic defects responsible for EB by introducing functional copies of mutated genes or modifying existing genes to restore protein function. Particularly for severe subtypes like Recessive Dystrophic Epidermolysis Bullosa (RDEB), gene therapy holds significant potential. This review aims to evaluate the role of new therapeutic approaches in the treatment of EB. The review includes findings from studies conducted on humans. While early studies and clinical trials have shown promising results, further research and trials are necessary to establish the safety and efficacy of these innovative approaches for EB treatment.

First report of Calonectria ilicicola causing red crown rot of soybean in Indiana.

Soybean (Glycine max [L.] Merr.) samples from commercial fields in Decatur and Spencer counties, Indiana were submitted to the Purdue Plant and Pest Diagnostic Lab in August to October 2022. Plants exhibited whole-leaf to interveinal chlorosis of the foliage, red to dark brown external lesions on the crown spreading from the soil-line upward, and severe root rot. In the fields, patches of diseased plants were observed, with greater than 50% of the plants affected and yield loss up to 50%. Orange to red perithecia were present on the exterior of symptomatic stem tissue and ranged in size from 329 to 433 × 232 to 306 µm (n = 10). Stems were surface sterilized in 10% Clorox (0.825% NaOCl) for 1 min, then rinsed with sterile distilled water and dried. In a laminar flow hood, sections of symptomatic stem tissue were plated on using quarter-strength potato dextrose agar (QPDA) and incubated under fluorescent lights on a 12-hr light/dark cycle at 20°C. After 6 days, fungal colonies with fluffy aerial hyphae, which were white near the colony margins and orange to burnt-red near their center, grew uniformly from the stem tissue plated. Elongate, cylindrical hyaline conidia with zero to three septations measuring 45.5 to 73.8 × 4.4 to 6.7 µm (n = 22) grew in clusters from symptomatic stem tissue within the plate. Perithecia developed after 14 days. Falcate, hyaline ascospores with one to two septa measuring 29.4 to 54.7 × 4.6 to 6.8 (n = 23) µm developed within the perithecia. Calonectria ilicicola Boedijn & Reitsma was confirmed based on morphological characteristics (Padgett et al. 2015). Isolate PPDL 22-01457B was used for DNA extraction using the ZR Fungal/Bacterial DNA Miniprep kit (Zymo Research, Irvine, CA). The internal transcriber region (ITS), actin (ACT) and β-tubulin (TUB2) genes were amplified (Carbone and Kohn 1999; Glass and Donaldson 1995; O'Donnell and Cigelnik 1997; White et al. 1990). Amplicons were sent for Sanger sequencing (Genewiz, Inc., South Plainfield, NJ), submitted to Genbank, and assigned accession numbers ITS: OQ932995, Actin: OR484986, and β-tubulin: OR546281. Sequences were analyzed using the NCBI BLASTn tool with results showing 99.5 to 100% identical to C. ilicicola (GenBank accessions LC500063, OQ303403, CP085825, respectively). To perform Koch's Postulates, 90 soybean seeds (CP3620E) were planted in potting media (Berger, Saint-Modeste, Quebec, Canada) in a seed flat with 45 of the plants used as controls and grown under grow lights for 16hr light/8hr dark at 20℃. Individual seedling crowns were inoculated 3 days post-emergence with a 5 to 10 ml spore and hyphal suspension that was scraped from the surface of a 14-day old QPDA culture after adding 300 mL deionized (DI) to each plate grown at 20 to 22°C. The control plants received sterile-DI water. Plants were covered in a plastic bag for 72 h. Plant stems were sprayed with sterile-DI water once a day for seven days. Symptoms were observed after four days, but significant crown rot and lesions developed after two weeks before wilting and dying. Calonectria iliciola was isolated uniformly from symptomatic plants and identified morphologically. Control plants showed no symptoms. Inoculations were repeated 3 times with similar results. As of fall 2023, red crown rot has been confirmed in Adams and Rush counties in Indiana. Red crown rot has been confirmed in several Midwest states (Kleczewski et al. 2019, Neves et al. 2023), but the extent of its distribution and disease management strategies are still limited.