First report of dasheen mosaic virus infecting calla lilies in South Korea.

In April 2022, leaves showing virus-like symptoms including mosaic, feathery chlorotic mottle and distortions were observed on calla lilies (Zantedeschia sp.) growing in a greenhouse in Jeolla province, South Korea. Leaf samples from nine symptomatic plants from the same greenhouse were collected and tested for Zantedeschia mosaic virus (ZaMV), Zantedeschia mild mosaic virus (ZaMMV) and Dasheen mosaic virus (DaMV) by reverse transcription-polymerase chain reaction (RT-PCR) with specific primers, ZaMV-F/R (Wei et al. 2008), ZaMMV-F/R (5'-GACGATCAGCAACAGCAGCAACAGCAGAAG-3'/5'-CTGCAAGGCTGAGATCCCGAGTAGCGAGTG-3') and DsMV-CPF/CPR, respectively. In previous surveys, ZaMV and ZaMMV were detected in calla lily fields in South Korea. Of 9 symptomatic samples, 8 were positive for ZaMV and ZaMMV but no PCR product was obtained from the ninth sample, which showed a yellow feather-like pattern. To identify the causal virus, total RNA from a leaf sample of the symptomatic calla lily was extracted using an RNeasy Plant Mini Kit (Qiagen, Germany) and analyzed by high-throughput sequencing. Ribosomal RNA was removed and a cDNA library was prepared using an Illumina TruSeq Stranded Total RNA LT Sample Prep Kit (Plants) and sequenced on an Illumina NovaSeq 6000 system (Macrogen, Korea), yielding 150 nt paired end reads. De novo assembly of the 88,171,036 reads was performed using Trinity software (r20140717) while the 113,140 initially assembled contigs were screened against the NCBI viral genome database using BLASTN. One contig of 10,007 bp (GenBank LC723667) shared 79.89-87.08% nucleotide (nt) identities to the available genomes of other DsMV isolates including Colocasia esculenta isolates Et5 (MG602227, 87.08%; Ethiopia) and CTCRI-II-14 (KT026108, 85.32%; India), and a calla lily isolate (AJ298033, 84.95%; China). No contigs representing other plant viruses were identified. To confirm the presence of DsMV, and because the virus was not detected using DsMV-CPF/CPR, RT-PCR was performed using new virus-specific primers DsMV-F/R (5'-GATGTCAACGCTGGCACCAGT-3'/5'-CAACCTAGTAGTAACGTTGGAGA-3'), designed based on the contig sequence. PCR products of the expected 600 bp were obtained from the symptomatic plant, cloned into the pGEM-T Easy Vector (Promega, USA), and two independent clones were bidirectionally sequenced (BIONEER, Korea), and shown to be identical. The sequence was deposited in GenBank as acc. no. LC723766, and shared 100% nt identity to the full-length contig LC723667, and 91.83% identity to the Chinese calla lily DsMV isolate (AJ298033). DsMV, a member of the genus Potyvitus in the family Potyviridae, is one of the major viruses infecting taro in South Korea, showing mosaic and chlorotic feathering symptoms (Kim et al. 2004); however, there is no record in the literature of the identification of this virus in South Korea in ornamental species including calla lily. To survey the sanitary status of other calla lilies, 95 samples with or without symptoms were collected from other regions and subjected to RT-PCR detection for DsMV. Ten of these samples were positive with primers DsMV-F/R, including seven mixed infections (DsMV+ZaMV or DsMV+ZaMV+ZaMMV). To our knowledge, this is the first report of DsMV infecting calla lilies in South Korea. The virus is easily spread by vegetative propagation (Babu et al. 2011) and by aphids (Reyes et al. 2006). This study will help the management of viral diseases on calla lilies in South Korea.

CD10 inhibits cell motility but expression is associated with advanced stage disease in colorectal cancer.

INTRODUCTION: CD10 is a cell membrane-bound endopeptidase which is expressed in normal small bowel but not in normal colon. It is aberrantly expressed in a small proportion of colorectal cancers (CRC) and this has been associated with liver metastasis and poor prognosis. We sought to investigate the mechanism of CD10 activity and its association with clinicopathological features. MATERIAL AND METHODS: CD10 was stably knocked down by lentiviral shRNA transduction in the CRC cell lines SW480 and SW620 which are derived from a primary tumour and its corresponding metastasis respectively. Expression of epithelial - mesenchymal transition (EMT) markers was tested as well as the effect of knockdown on cell viability, migration and invasion assays. In addition, immunohistochemical expression of CD10 in primary colorectal tumours (N = 84) in a tissue microarray was digitally quantified and analysed for associations with clinicopathological variables. RESULTS: Knockdown of CD10 did not alter cell viability in SW480, but migration and invasion levels increased (P < 0.001 for each) and this was associated with a cadherin switch. In SW620, CD10 knockdown caused a reduction in cell viability after 72 h (P = 0.0018) but it had no effect on cell migration and invasion. Expression of epithelial CD10 in primary tumours was associated with presence of lymph node invasion (P = 0.001) and advanced Duke's stage (P = 0.001). CONCLUSIONS: Our results suggest that the function of CD10 may change during tumour evolution. It may inhibit cell motility in early-stage disease whilst promoting cell viability in late-stage disease. It has a complex role and further studies are needed to elucidate the suitability of CD10 as a prognostic marker or therapeutic target.

Influence of potassium concentration gradient on stable caesium uptake by Calla palustris.

The effect of potassium (K) concentration gradient on stable caesium (Cs) uptake by Calla palustris was studied under hydroponic conditions after eight-day exposure in a greenhouse experiment. The plants were exposed to two different concentrations of Cs (provided as 0.5 and 1 mM CsCl) and five different concentrations of K (provided as K2SO4 in 0.5, 1, 2, 5, and 10 mM). The results indicate negative dependence of Cs uptake on K concentrations for both Cs treatments. The application of K reduced the transfer of stable Cs from water to plant by about 44-72% for 0.5 mM CsCl and 56-74% for 1 mM CsCl. The highest efficiency of Cs removal from water was observed for plants in K+ deficient solutions (plants starving), with an efficiency 8.0% for plants cultivated in 0.5 mM CsCl and 9.4% for plants in 1 mM CsCl. An increasing concentration of K also supported translocation of Cs from roots to leaves. Higher translocation was observed for the treatments with lower level of Cs, where the concentration of Cs in leaves became higher than that in roots. The Cs uptake and translocations were affected not only by the external concentration of K, but also the external concentration of stable Cs. A high concentration of K in the environment protects the food chain from Cs uptake by plants, but lowers the efficiency of phytoremediation techniques.

Stable cesium (133Cs) uptake by Calla palustris from different substrates.

The uptake of stable cesium (133Cs) by Calla palustris was evaluated from four different substrates: water, soil, keramzit (a clay granule) and water with the addition of a potassium compound, after an eight days exposure to a solution of 0.5mM cesium chloride. Stable cesium was used because it is commonly supposed that its uptake by plants is the same of that of radiocesium (137Cs). The plants were differentiated in their parts (roots, healthy leaves, dead leaves and flowers) and analyzed with ICP-MS. The lowest average concentration of absorbed Cs was found in plants exposed in soil (0.7mg/kg, S.D.=96.8), while the highest in plants exposed in water (147mg/kg, S.D.=51.7). During the experiment the water planted plants removed 31.6% of provided Cs while those planted in soil removed only 0.06%. The addition of potassium to water was tested because of the competition effect that arises between these two elements: this effect was confirmed with the result that the average uptake in the presence of potassium was lower (41mg/kg in exposed plants, S.D.=76.1). The uptake was also lower in the solid-based substrates (soil and keramzit), because of the known tendency of Cs to bind with soil particles, thus becoming less available to plants. There was no evidence that the different parts of the plant showed different uptake effectiveness, or that the health of the plant (evaluated with a qualitative method) had any effect on the uptake of Cs.

Develop a preliminary core germplasm with the novel polymorphism EST-SSRs derived from three transcriptomes of colored calla lily (Zantedeschia hybrida).

The development of high-throughput sequencing technology has made it possible to develop molecular markers such as EST-SSR from transcriptome sequences in non-model plants such as bulbous flowers. However, the EST-SSR markers that have been developed are weakly validated and low polymorphic due to the short read size and poor quality of the assembled sequences. This study therefore used the CandiSSR pipeline to identify 550 potential polymorphic SSR loci among 487 homologous unigenes based on the transcriptomic sequences of three varieties of colored calla lily, and 460 of these loci with appropriate flanking sequences were suitable for primer pairs design. A further validation with 200 randomly selected EST-SSRs demonstrated an increase of more than 30% and 100% in amplification validity and polymorphism, respectively, in comparison with our previous study. In addition, since most of the current varieties of colored calla lily are hybridized from a few species, which have low genetic diversity, we subsequently identified primary core germplasm for 160 colored calla lily accessions using the aforementioned 40 polymorphic EST-SSRs. It was concluded that the core germplasm containing 42 accessions derived from the M strategy incorporated into the software Power Core was the most representative of all 160 original germplasm, as evidenced by the preservation of 100% of the EST-SSR variation, with a higher level of genetic diversity and heterogeneity (Nei = 0.40, I = 0.66, PIC = 0.43). This study provides a practical example of polymorphism EST-SSR markers developed from multiple transcriptomes for non-model plants. A future breeding program for colored calla lily will also benefit from the core germplasm defined by those molecular markers.

Erythrophagocytosis in a patient with B-cell acute lymphoblastic leukemia with t(12;21) (p13.2; q22.1); ETV6-RUNX1: Case report and review of the literature.

Erythrophagocytosis (EP) is extremely rare in de novo acute lymphoblastic leukemia (ALL). We document a rare case, which in addition, showed extensive blast vacuolization. A detailed literature review has also been incorporated with the aim of unraveling the prognostic import of our morphological observations if any. A five-year-old male presented with fever and progressive pallor for 1 month. He had hepatosplenomegaly and bicytopenia. Peripheral smear examination showed 43% blasts. Nuclear and cytoplasmic vacuolations were seen in 75% blasts and EP in 4% blasts. The blasts showed block positivity on periodic acid-Schiff (PAS) stain. Marrow aspirate smears showed 58% blasts displaying a similar morphology. Flow cytometry showed features of a common acute lymphoblastic leukemia antigen (CALLA) positive B-cell ALL with aberrant, dim CD 33 expression in 53.4% of the gated blasts. Fluorescence in situ hybridization showed translocation (12; 21) (p13;q22). The patient responded well to standard induction therapy. To conclude, EP is rarely seen in de novo ALL and is associated with a favorable translocation, t(12;21).

Establishment of an efficient regeneration and Agrobacterium transformation system in mature embryos of calla lily (Zantedeschia spp.).

Calla lily (Zantedeschia spp.) have great aesthetic value due to their spathe-like appearance and richness of coloration. However, embryonic callus regeneration is absent from its current regeneration mechanism. As a result, constructing an adequate and stable genetic transformation system is hampered, severely hindering breeding efforts. In this research, the callus induction effectiveness of calla lily seed embryos of various maturities was evaluated. The findings indicated that mature seed embryos were more suitable for in vitro regeneration. Using orthogonal design experiments, the primary elements influencing in vitro regeneration, such as plant growth regulators, genotypes, and nanoscale materials, which was emergent uses for in vitro regeneration, were investigated. The findings indicated that MS supplemented with 6-BA 2 mg/L and NAA 0.1 mg/L was the optimal medium for callus induction (CIM); the germination medium (GM) was MS supplemented with 6-BA 2 mg/L NAA 0.2 mg/L and 1 mg/L CNTs, and the rooting medium (RM) was MS supplemented with 6-BA 2 mg/L NAA 0.7 mg/L and 2 mg/L CNTs. This allowed us to verify, in principle, that the Agrobacterium tumefaciens-mediated genetic transformation system operates under optimal circumstances using the GUS reporter gene. Here, we developed a seed embryo-based genetic transformation regeneration system, which set the stage for future attempts to create new calla lily varieties.

Leaf NPK stoichiometry, δ15 N, and apparent nutrient limitation of co-occurring carnivorous and noncarnivorous plants.

Previous meta-analyses suggested that carnivorous plants-despite access to N, P, and K from prey-have significantly lower leaf concentrations of these nutrients than noncarnivores. Those studies, however, largely compared carnivores in nutrient-poor habitats with noncarnivores in more nutrient-rich sites, so that the differences reported might reflect habitat differences as much as differences in nutrient-capture strategy. Here we examine three carnivorous and 12 noncarnivorous plants in the same nutrient-poor bog to compare their foliar nutrient concentrations, assess their patterns of nutrient limitation using leaf NPK stoichiometry, and estimate percentage N derived from prey by carnivores using a mixing model for stable N isotopes. We hypothesized that (1) carnivore leaf nutrient concentrations approach or exceed those of noncarnivores in the same nutrient-poor habitat; (2) species in different functional groups show different patterns of stoichiometry and apparent nutrient limitation; and (3) noncarnivores might show evidence of using other means of nutrient acquisition or conservation to reduce nutrient limitation. At Fallison Bog in northern Wisconsin, carnivorous plants (Drosera rotundifolia, Sarracenia purpurea, Utricularia macrorhiza) showed significantly lower leaf percentage C and N:P ratio, higher δ15 N, and no difference from noncarnivores in leaf N, P, K, and δ13 C. Sedges had significantly lower leaf percentage P, percentage C, and N:K ratio, and higher K:P ratio than nonsedges restricted to the Sphagnum mat, and may tap peat N via aerenchyma-facilitated peat oxidation (oxipeditrophy). Evergreen ericaceous shrubs exhibited significantly higher levels of percentage C and lower values of δ15 N than mat nonericads. Calla palustris-growing in the nutrient-rich moat at the bog's upland edge-had very high values of leaf N, K, δ15 N, and N:P ratio, suggesting that it may obtain nutrients from minerotrophic flows from the adjacent uplands and/or rapidly decaying peat. Stoichiometric analyses indicated that most species are N limited. A mixing model applied to δ15 N values for carnivores, noncarnivores, and insects produced an estimate of 50% of leaf N derived from prey for Utricularia, 42% for Sarracenia, and 41% for Drosera.

Neprilysin expression and functions in development, ageing and disease.

Neprilysin (NEP) is an integral membrane-bound metallopeptidase with a wide spectrum of substrates and physiological functions. It plays an important role in proteolytic processes in the kidney, cardiovascular regulation, immune response, cell proliferation, foetal development etc. It is an important neuropeptidase and amyloid-degrading enzyme which makes NEP a therapeutic target in Alzheimer's disease (AD). Moreover, it plays a preventive role in development of cancer, obesity and type-2 diabetes. Recently a role of NEP in COVID-19 pathogenesis has also been suggested. Despite intensive research into NEP structure and functions in different organisms, changes in its expression and regulation during brain development and ageing, especially in age-related pathologies, is still not fully understood. This prevents development of pharmacological treatments from various diseases in which NEP is implicated although recently a dual-acting drug sacubitril-valsartan (LCZ696) combining a NEP inhibitor and angiotensin receptor blocker has been approved for treatment of heart failure. Also, various natural compounds capable of upregulating NEP expression, including green tea (EGCG), have been proposed as a preventive medicine in prostate cancer and AD. This review summarizes the existing literature and our own research on the expression and activity of NEP in normal brain development, ageing and under pathological conditions.

Characterization of Pectobacterium carotovorum proteins differentially expressed during infection of Zantedeschia elliotiana in vivo and in vitro which are essential for virulence.

The identification of phytopathogen proteins that are differentially expressed during the course of the establishment of an infection is important to better understand the infection process. In vitro approaches, using plant extracts added to culture medium, have been used to identify such proteins, but the biological relevance of these findings for in planta infection are often uncertain until confirmed by in vivo studies. Here, we compared the proteins of Pectobacterium carotovorum ssp. carotovorum strain PccS1 differentially expressed in Luria-Bertani medium supplemented with extracts of the ornamental plant Zantedeschia elliotiana cultivar 'Black Magic' (in vitro) and in plant tissues (in vivo) by two-dimensional electrophoresis coupled with mass spectrometry. A total of 53 differentially expressed proteins (>1.5-fold) were identified (up-regulated or down-regulated in vitro, in vivo or both). Proteins that exhibited increased expression in vivo but not in vitro, or in both conditions, were identified, and deletions were made in a number of genes encoding these proteins, four of which (clpP, mreB, flgK and eda) led to a loss of virulence on Z. elliotiana, although clpP and mreB were later also shown to be reduced in growth in rich and minimal media. Although clpP, flgK and mreB have previously been reported as playing a role in virulence in plants, this is the first report of such a role for eda, which encodes 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase, a key enzyme in Entner-Doudoroff metabolism. The results highlight the value of undertaking in vivo as well as in vitro approaches for the identification of new bacterial virulence factors.

Assessing Genetic Diversity and Population Differentiation of Colored Calla Lily (Zantedeschia Hybrid) for an Efficient Breeding Program.

Plastome-genome incompatibility (PGI) is prevalent in several plants including the Zantedeschia species, a worldwide commercial flower crop native to South Africa. Generally, hybrids suffering from PGI appear less vigorous and more susceptible than normal plants. Previous reports revealed that the PGI level in interspecific hybrids is correlated with the relatedness of the parental species in the genus Zantedeschia. To provide a basis for utilizing and improving resources in breeding programs, a total of 117 accessions of colored calla lily (Zantedeschia hybrid), collected from New Zealand, the Netherlands and the United States, were genotyped using 31 transferable expressed sequence tags-simple sequence repeats (EST-SSR) markers from the white calla lily (Zantedeschia aethiopica). A moderately high level of genetic diversity was observed, with 111 alleles in total, an observed/expected heterozygosity (Ho/He) of 0.453/0.478, and polymorphism information content (PIC) of 0.26. Genetic distance and STRUCTURE-based analysis further clustered all accessions into four subgroups (G-Ia, G-Ib, G-IIa and G-IIb), which mostly consisted of Zantedeschia pentlandii, Zantedeschia elliotiana, Zantedeschia albomaculata and Zantedeschia rehmannii, respectively. Significant genetic differentiation was observed between all inferred subgroup pairs, with the Fst ranging from 0.142 to 0.281. Finally, the accessions assigned into G-IIb (Z. rehmannii) were recommended as top priority parents in efficient Zantedeschia breeding program designs.

Transcriptome analysis of colored calla lily (Zantedeschia rehmannii Engl.) by Illumina sequencing: de novo assembly, annotation and EST-SSR marker development.

Colored calla lily is the short name for the species or hybrids in section Aestivae of genus Zantedeschia. It is currently one of the most popular flower plants in the world due to its beautiful flower spathe and long postharvest life. However, little genomic information and few molecular markers are available for its genetic improvement. Here, de novo transcriptome sequencing was performed to produce large transcript sequences for Z. rehmannii cv. 'Rehmannii' using an Illumina HiSeq 2000 instrument. More than 59.9 million cDNA sequence reads were obtained and assembled into 39,298 unigenes with an average length of 1,038 bp. Among these, 21,077 unigenes showed significant similarity to protein sequences in the non-redundant protein database (Nr) and in the Swiss-Prot, Gene Ontology (GO), Cluster of Orthologous Group (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Moreover, a total of 117 unique transcripts were then defined that might regulate the flower spathe development of colored calla lily. Additionally, 9,933 simple sequence repeats (SSRs) and 7,162 single nucleotide polymorphisms (SNPs) were identified as putative molecular markers. High-quality primers for 200 SSR loci were designed and selected, of which 58 amplified reproducible amplicons were polymorphic among 21 accessions of colored calla lily. The sequence information and molecular markers in the present study will provide valuable resources for genetic diversity analysis, germplasm characterization and marker-assisted selection in the genus Zantedeschia.

CD10-(CALLA)-Positive Lymphocytes in Myeloma: Evidence that they are a Malignant Precursor Population and are of Germinal Centre Origin.

CD10 antigen has been repeatedly detected on putative lymphoid precursor populations in both the bone marrow and circulation of multiple myeloma patients, as well as on the plasma cells in some cases of myeloma. The presence of these CD10-positive cells has raised questions regarding the ontogeny of the proliferating precursor cell in myeloma. The majority opinion has implicated a CD10-positive haemopoietic progenitor cell. However, the CD10 antigen has been detected on some mature B cells, i.e. germinal centre B cells. In this paper we postulate that the proliferating precursor cell in myeloma arises from the germinal centre. The germinal centre is the site of affinity maturation of antibody responses via somatic mutation and of isotype switching. Thus the siting of the clonogenic cell in myeloma in the germinal centre explains the overwhelming predominance of IgG and IgA myelomas, the phenomenon of point mutation which occurs in myeloma proteins in the presence of stable immunoglobulin gene rearrangements and the impaired primary immune response in myeloma. It is also consistent with the requirement for antigenic exposure in the development of myelomatosis.

Aggressive Diffuse Lymphoma Coexpressing NRAS p21 and C-erbB-2 (neu) Oncogene Products, and CALL A (CD 10).

We describe here a case of malignant lymphoma (ML) which coexpressed common acute lymphoblastic leukemia antigen (CALLA:CD10) and NRAS p21 and c-erbB-2 (neu) oncogene products. The patient, an 83 year-old man, had massive generalized lymphadenopathy and pleural effusions. Serum LDH levels were elevated to 801 IU/L. Surface phenotypes were analysed by a fluorescent-activated cell sorter with a panel of monoclonal antibodies (MAbs). The ML cells coexpressed antigens detected by MAbs CD10, CD19, CD20, CD22, CD24, CD38, Ia (HLA-DR), c-neu and surface immunoglobulin (Ig) G, Kappa. Gene rearrangements for the Ig JH and JK were found. Overexpression of NRAS p21 was shown by gene amplification using Southern blot analysis, while gene amplification of c-erbB-2 oncogene was also demonstrated. To our knowledge, this is the first report to demonstrate an overexpression of p185 c-neu on ML cells. These findings suggest that the p185 neu may be a prognostic indicator not only for breast adenocarcinomas but also for lymphoproliferative disorders, and that the transforming p185 protein may be involved in the mechanisms of aggressive expansion of lymphoid neoplasias.

Distinctive Pattern of Expression of Activation and Resting B Cell Antigens on Normal and Neoplastic Human B Cells: Immunophenotypic Heterogeneity in Some Lymphomas.

The pattern of expression of four B cell antigen systems on mature human B cells and B cell lymphomas were studied. The L30 antigen was detected on small resting B cells, while the B cell activation antigens, CD10, CD25 and L29, were expressed differentially on activated B cells. The multiparameter-flowcytometric analysis of these four antigens revealed that mature B cells changed their pattern of expression in an activation-stage specific manner. Thus, the presence of L30, CD10, CD25 and L29 on mature human B cells correlated with distinct B cell populations at a particular stage of activation. Histo-pathologically well defined B cell lymphomas were also studied for the expression of these four antigens. Burkitt's lymphoma and diffuse small cleaved lymphoma were found to have an heterogeneous expression of these antigens, suggesting that certain types of non-Hodgkin's lymphoma (NHL) are immunophenotypically heterogeneous, and that this heterogeneity may reflect a different biology and behavior in vivo.