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.

Plant growth-promoting bacteria (PGPB) are increasingly deployed as biofertilizers, yet the link between an inoculant’s genomic potential and its realized effect on the plant is rarely assessed within an integrative framework that jointly captures the rhizosphere microbiome, plant phenotype, and strain genome. Two Bacillus strains—B. halotolerans 1453 and B. pumilus 630—were applied to wheat and soybean in a factorial pot experiment (2 strains × 2 application methods × 3 frequencies + control, 3–4 replicates). Rhizosphere samples (n = 67 after filtering) were profiled by 16S rRNA sequencing with PICRUSt2 functional prediction and compositional validation (Aitchison PERMANOVA, ALDEx2, ANCOM-BC2). The PGPB gene repertoire was characterized by genome mining (481 marker genes, 14 categories). Wheat phenotype (six traits) and soybean height were analyzed with models appropriate for count data (Negative Binomial and binomial GLMs) for treatment-vs.-control comparisons, and with factorial ANOVA for decomposition into main effects and interactions. Crop identity was the dominant factor shaping both microbiome structure and function (PERMANOVA R² = 14.7% taxonomically and R² = 7.8% functionally, both p < 0.001), with biologically meaningful taxonomic differences between wheat and soybean; strain, application count and method had no significant effect on community composition (R² < 4% each), and co-occurrence networks showed no reliable differences between crops once read depth and sample size were controlled for. Despite this neutrality at the microbiome level, inoculation significantly increased wheat spike count (NB-GLM, all 12 treatments vs. control, padj 0.0002–0.031), ear weight, and stem count, with application count the strongest source of variability and a pronounced strain × application count. Strain 1453 outperformed 630 in spike count (+23.1%, p = 0.012) and ear weight (+20.4%, p = 0.023); we hypothesize that this may be related to its more complete DNRA pathway (narGHI + nirB-nirD) and biocontrol genes (bacE, srfAA). Strain 630 produced a less pronounced effect than strain 1453 but was subject to smaller fluctuations across replicates (CV ≈ 16–21% vs. ≈24–26% for 1453), which may reflect better resilience to environmental fluctuations, possibly due to its confirmed rsbV/rsbW stress-tolerance regulon. Rhizosphere microbiome composition differed clearly by crop (wheat vs. soybean) but showed no detectable response to strain, application method, or application count. Despite this lack of a microbiome signal, inoculation significantly increased wheat spike count and ear weight, with the magnitude and stability of this effect differing by strain. We hypothesize that this strain-dependent difference relates to underlying genomic differences—particularly in nitrogen metabolism (DNRA pathway) and stress-tolerance genes—though this link has not been tested directly and remains a hypothesis for future work.

Introduction. Cerebral small vessel disease (SVD) is an early but nonspecific manifestation of Fabry disease. We describe a middle-aged patient with severe SVD in whom recognition of long-standing systemic disease ultimately led to the detection of a likely pathogenic GLA variant and enabled initiation of enzyme replacement therapy before advanced renal involvement developed. Methods. The patient underwent clinical evaluation, brain MRI, whole-genome sequencing, biochemical analysis of dried blood spots, and Sanger sequencing. Results. A 41-year-old man presented with recurrent lacunar strokes, progressive gait impairment, pseudobulbar syndrome, cognitive decline, and bladder dysfunction. Brain MRI revealed severe SVD with confluent white matter hyperintensities, multiple lacunes, and microbleeds. From early adulthood, he had arterial hypertension, hypohidrosis, acroparesthesias, and mild proteinuria. Later, he developed a cataract and an acute dissection of the ascending aorta. Whole-genome sequencing identified a hemizygous GLA frameshift variant (c.1095dup), predicted to truncate alpha-galactosidase A (p.Thr366TyrfsTer9) and classified as likely pathogenic. The diagnosis was confirmed by Sanger sequencing, markedly reduced alpha-galactosidase A activity, and elevated lyso-Gb3 in dried blood spots. Discussion. This case expands the neurologic phenotype of Fabry disease and supports its inclusion in the differential diagnosis of severe early-onset SVD, even when the classic systemic phenotype is incomplete.

Nitrogen deficiency is one of the key factors limiting the growth and raw-material quality of tea plants; however, cell-autonomous mechanisms underlying the response of Camellia sinensis (L.) Kuntze to reduced nitrogen availability remain insufficiently studied. This study characterized biochemical and transcriptomic changes in callus cultures of three tea cultivars (‘Kolkhida’, ‘Karatum’, and cultivar #582) after two months of cultivation on Murashige–Skoog medium lacking nitrogen-containing components. Nitrogen withdrawal caused visible stress symptoms, reduced total nitrogen in ‘Kolkhida’ and ‘Karatum’ (total nitrogen was not determined in cultivar #582), and altered the accumulation of L-theanine, caffeine, and catechins in a cultivar-dependent manner. ‘Kolkhida’ showed decreases in simple catechins, L-theanine, and caffeine; ‘Karatum’ showed increases in simple and gallated catechins together with a decrease in L-theanine; and cultivar #582 showed smaller changes in most measured biochemical traits. RNA-seq identified 1898 differentially expressed genes (DEGs) in ‘Kolkhida’, 4551 in ‘Karatum’, and 9869 in cultivar #582. The enriched Gene Ontology term “response to karrikin” (GO:0080167) was common to all three cultivars, whereas cultivar-specific profiles differed substantially. In the flavonoid biosynthesis pathway, key phenylpropanoid genes were predominantly downregulated in cultivar #582 but induced in ‘Kolkhida’ and ‘Karatum’. RT-qPCR confirmed the direction of change for selected genes. Under the tested in vitro conditions, ‘Kolkhida’ showed the strongest biochemical deterioration, whereas cultivar #582 combined the greatest biochemical stability with the most extensive transcriptomic remodeling. Because total nitrogen was not measured in cultivar #582, its apparent tolerance remains provisional. Tea callus cultures therefore provide a controlled discovery system for cell-autonomous candidate responses, but cultivar rankings and candidate markers require validation in independent callus lines and whole plants.

Nitrogen (N) availability strongly influences tea growth and the accumulation of quality-related metabolites, but long-term responses to withdrawal of external N can vary among cultivars. In this study, we characterized phenotypic, spectral, biochemical, and transcriptomic responses of four Camellia sinensis accessions—‘Kolkhida’, ‘Karatum’, and γ-irradiation-derived mutant forms #582 and #619—during two, four, or six months of complete withdrawal of exogenous NH₄NO₃ in greenhouse sand culture. Phenotypic, spectral, and biochemical measurements included all four accessions at all three time points; RNA-seq data were available for all four accessions were represented at four and six months, whereas two-month point were available only for ‘Kolkhida’ and ‘Karatum’. Nitrogen withdrawal decreased L-theanine and caffeine and generally increased simple and gallated catechins, consistent with a change in carbon/nitrogen balance towards phenolic metabolism. The four cultivars exhibited markedly distinct responses. ‘Karatum’ showed the optimum resilience, with minimal phenotypic alteration and a neutral transcriptional response, while #582 demonstrated a stronger stress-associated phenotype and metabolic response. Within the available RNA-seq comparisons, DEG counts were lower at four months than at six months, while the two-month datasets for ‘Kolkhida’ and ‘Karatum’ contained the largest DEG sets for those two cultivars, suggesting a phased acclimation process. CsELIP1, CsSRG1, CsHHO2/4, CsNRT2.4, and CsTAT2 were identified as potential candidate genes associated with nitrogen limitation, flavonoid regulation, and amino acid metabolism. Our findings show that N-deficiency reactions in tea are highly cultivar- and time-dependent, making single-time-point evaluations insufficient. We propose ‘Karatum’ as a promising candidate for further evaluation in low-nitrogen dose–response and field trials based on our combined biochemical and transcriptional findings.