Mechanisms underlying the formation of complex color patterns on Nigella orientalis (Ranunculaceae) petals.

Complex color patterns on petals are widespread in flowering plants, yet the mechanisms underlying their formation remain largely unclear. Here, by conducting detailed morphological, anatomical, biochemical, optical, transcriptomic, and functional studies, we investigated the cellular bases, chromogenic substances, reflectance spectra, developmental processes, and underlying mechanisms of complex color pattern formation on Nigella orientalis petals. We found that the complexity of the N. orientalis petals in color pattern is reflected at multiple levels, with the amount and arrangement of different pigmented cells being the key. We also found that biosynthesis of the chromogenic substances of different colors is sequential, so that one color/pattern is superimposed on another. Expression and functional studies further revealed that a pair of R2R3-MYB genes function cooperatively to specify the formation of the eyebrow-like horizontal stripe and the Mohawk haircut-like splatters. Specifically, while NiorMYB113-1 functions to draw a large splatter region, NiorMYB113-2 functions to suppress the production of anthocyanins from the region where a gap will form, thereby forming the highly specialized pattern. Our results provide a detailed portrait for the spatiotemporal dynamics of the coloration of N. orientalis petals and help better understand the mechanisms underlying complex color pattern formation in plants.

Machine learning for differentiation of lipid-poor adrenal adenoma and subclinical pheochromocytoma based on multiphase CT imaging radiomics.

BACKGROUND: There is a paucity of research investigating the application of machine learning techniques for distinguishing between lipid-poor adrenal adenoma (LPA) and subclinical pheochromocytoma (sPHEO) based on radiomic features extracted from non-contrast and dynamic contrast-enhanced computed tomography (CT) scans of the abdomen. METHODS: We conducted a retrospective analysis of multiphase spiral CT scans, including non-contrast, arterial, venous, and delayed phases, as well as thin- and thick-thickness images from 134 patients with surgically and pathologically confirmed. A total of 52 patients with LPA and 44 patients with sPHEO were randomly assigned to training/testing sets in a 7:3 ratio. Additionally, a validation set was comprised of 22 LPA cases and 16 sPHEO cases from two other hospitals. We used 3D Slicer and PyRadiomics to segment tumors and extract radiomic features, respectively. We then applied T-test and least absolute shrinkage and selection operator (LASSO) to select features. Six binary classifiers, including K-nearest neighbor (KNN), logistic regression (LR), decision tree (DT), random forest (RF), support vector machine (SVM), and multi-layer perceptron (MLP), were employed to differentiate LPA from sPHEO. Receiver operating characteristic (ROC) curves and area under the curve (AUC) values were compared using DeLong's method. RESULTS: All six classifiers showed good diagnostic performance for each phase and slice thickness, as well as for the entire CT data, with AUC values ranging from 0.706 to 1. Non-contrast CT densities of LPA were significantly lower than those of sPHEO (P < 0.001). However, using the optimal threshold for non-contrast CT density, sensitivity was only 0.743, specificity 0.744, and AUC 0.828. Delayed phase CT density yielded a sensitivity of 0.971, specificity of 0.641, and AUC of 0.814. In radiomics, AUC values for the testing set using non-contrast CT images were: KNN 0.919, LR 0.979, DT 0.835, RF 0.967, SVM 0.979, and MLP 0.981. In the validation set, AUC values were: KNN 0.891, LR 0.974, DT 0.891, RF 0.964, SVM 0.949, and MLP 0.979. CONCLUSIONS: The machine learning model based on CT radiomics can accurately differentiate LPA from sPHEO, even using non-contrast CT data alone, making contrast-enhanced CT unnecessary for diagnosing LPA and sPHEO.

Direction of Arrival Estimation and Highlighting Characteristics of Testing Wideband Echoes from Multiple Autonomous Underwater Vehicles.

Multiple autonomous underwater vehicles (AUVs) have gradually become the trend in underwater operations. Identifying and detecting these new underwater multi-targets is difficult when studying underwater moving targets. A 28-element transducer is used to test the echo of multiple AUVs with different layouts in a lake. The characteristics of the wideband echo signals are studied. Under the condition that the direction of arrival (DOA) is not known, an autofocus coherent signal subspace (ACCSM) method is proposed. The focusing matrix is constructed based on the received data. The spatial spectrum of the array signal of multiple AUVs at different attitudes is calculated. The algorithm estimates the DOA of the echo signals to overcome the shortcomings of traditional wideband DOA estimation and improve its accuracy. The results show that the highlights are not only related to the number of AUVs, but are also modified by scale and attitude. The contribution of the microstructure of the target in the overall echo cannot be ignored. Different parts of the target affect the number of highlights, thus resulting in varying numbers of highlights at different attitude angle intervals. The results have significant implications for underwater multi-target recognition.

Association of a newly identified lncRNA LNC_000280 with the formation of acetylcholine receptor clusters in vitro.

Peripheral nerve injury (PNI) can disintegrate acetylcholine receptor (AChR) clusters in the postsynaptic membrane. In our previous research, lncRNAs that were differentially expressed in the whole transcriptome sequencing of denervated muscle atrophy after PNI were screened. By utilizing Gene Ontology (GO) analysis and protein-protein interaction (PPI) networks, a novel lncRNA LNC_000280 was predicted to be associated with neuromuscular junction (NMJ). The myotubes were used to assess the connection between LNC_000280 and AChR cluster formation in vitro by overexpression and knockdown of LNC_000280 in the C2C12 cell line. Our findings demonstrated that the overexpression of LNC_000280 repressed the gene expression and protein level of AChR subunits in myotubes and further reduced the area of AChR aggregates on the cell membrane. In contrast, the knockdown of LNC_000280 brought about opposite results. In addition, the transcriptional level of Sorbs2 changed inversely with the quantity change of LNC_000280. In conclusion, LNC_000280 may associate with the formation of AChR clusters.

Petal development and elaboration.

Petals can be simple or elaborate, depending on whether they have complex basic structures and/or highly specialized epidermal modifications. It has been proposed that the independent origin and diversification of elaborate petals have promoted plant-animal interactions and, therefore, the evolutionary radiation of corresponding plant groups. Recent advances in floral development and evolution have greatly improved our understanding of the processes, patterns, and mechanisms underlying petal elaboration. In this review, we compare the developmental processes of simple and elaborate petals, concluding that elaborate petals can be achieved through four main paths of modifications (i.e. marginal elaboration, ventral elaboration, dorsal elaboration, and surface elaboration). Although different types of elaborate petals were formed through different types of modifications, they are all results of changes in the expression patterns of genes involved in organ polarity establishment and/or the proliferation, expansion, and differentiation of cells. The deployment of existing genetic materials to perform a new function was also shown to be a key to making elaborate petals during evolution.

Identification of novel prognostic targets in glioblastoma using bioinformatics analysis.

BACKGROUND: Glioblastoma (GBM) is the most malignant grade of glioma. Highly aggressive characteristics of GBM and poor prognosis cause GBM-related deaths. The potential prognostic biomarkers remain to be demonstrated. This research builds up predictive gene targets of expression alterations in GBM utilizing bioinformatics analysis. METHODS AND RESULTS: The microarray datasets (GSE15824 and GSE16011) associated with GBM were obtained from Gene Expression Omnibus (GEO) database to identify the differentially expressed genes (DEGs) between GBM and non-tumor tissues. In total, 719 DEGs were obtained and subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) for function enrichment analysis. Furthermore, we constructed protein-protein Interaction (PPI) network among DEGs utilizing Search Tool for the Retrieval of Interacting Genes (STRING) online tool and Cytoscape software. The DEGs of degree > 10 was selected as hub genes, including 73 upregulated genes and 21 downregulated genes. Moreover, MCODE application in Cytoscape software was employed to identify three key modules involved in GBM development and prognosis. Additionally, we used the Gene expression profiling and interactive analyses (GEPIA) online tool to further confirm four genes involving in poor prognosis of GBM patients, including interferon-gamma-inducible protein 30 (IFI30), major histocompatibility complex class II-DM alpha (HLA-DMA), Prolyl 4-hydroxylase beta polypeptide (P4HB) and reticulocalbin-1 (RCN1). Furthermore, the correlation analysis indicated that the expression of IFI30, an acknowledged biomarker in glioma, was positively correlated with HLA-DMA, P4HB and RCN1. RCN1 expression was positively correlated with P4HB and HLA-DMA. Moreover, qRT-PCR and immunohistochemistry analysis further validated the upregulation of four prognostic markers in GBM tissues. CONCLUSIONS: Analysis of multiple datasets combined with global network information and experimental verification presents a successful approach to uncover the risk hub genes and prognostic markers of GBM. Our study identified four risk- and prognostic-related gene signatures, including IFI30, HLA-DMA, P4HB and RCN1. This gene sets contribute a new perspective to improve the diagnostic, prognostic, and therapeutic outcomes of GBM.

Neutrophil peptide-1 promotes the repair of sciatic nerve injury through the expression of proteins related to nerve regeneration.

Objectives Small-molecule polypeptide neutrophil peptide 1 (NP-1) was reported to promote the regeneration of the sciatic nerve after denervation, but the mechanisms underlying this effect of NP-1 are unclear. Here, we established a Sprague-Dawley rat model of crush injury to study the effect of a single intermuscular injection of NP-1 on the repair of injured peripheral nerves and elucidate the possible underlying mechanism.Methods 39 rats were randomly selected to join this study and divided into the blank control group (normal group, n=9), experimental group (NP-1 group, n=15), and negative control group (NS group, n=15). The dynamic expression of cytokines in different groups of nerve tissues during Wallerian degeneration was observed using protein chips at different time points after injury. Recovery of injured nerves was determined based on the general condition, local gross morphology of the nerve suture site, sciatic nerve function index, neuroelectrophysiology, and osmic acid staining at 6 weeks after the surgery. The recovery of effector function was determined based on wet weight, hematoxylin-eosin staining, modified Gomori staining, and nicotinamide adenine dinucleotide-tetrazolium reductase staining at 6 weeks after the surgery.Results It was found that a single topical administration of NP-1 promoted sciatic nerve regeneration after crush injury and affected the expression of proteins related to neurotrophy, inflammation, cell chemotaxis, and cell generation pathways.

Quantum Correlation Swapping between Two Werner States Undergoing Local and Nonlocal Unitary Operations.

In this paper, quantum correlation (QC) swapping between two Werner-like states, which are transformed from Werner states undergoing local and nonlocal unitary operations, are studied. Bell states measures are performed in the middle node to realize the QC swapping and correspondingly final correlated sates are obtained. Two different QC quantifiers, i.e., measurement-induced disturbance (MID) and ameliorated MID, are employed to characterize and quantify all the concerned QCs in the swapping process. All QCs in the concerned states are evaluated analytically and numerically. Correspondingly, their characteristics and properties are exposed in detail. It is exposed that, through the QC swapping process, one can obtain the long-distance QC indeed. Moreover, the similarities of monotony features of MID and AMID between the initial states and final states are exposed and analyzed.

Metabolomic profiling reveals serum L-pyroglutamic acid as a potential diagnostic biomarker for systemic lupus erythematosus.

OBJECTIVE: The spectrum of clinical manifestations and serological phenomena of SLE is heterogeneous among patients and even changes over time unpredictably in individual patients. For this reason, clinical diagnosis especially in complicated or atypical cases is often difficult or delayed leading to poor prognosis. Despite the medical progress nowadays in the understanding of SLE pathogenesis, disease-specific biomarkers for SLE remain an outstanding challenge. Therefore, we undertook this study to investigate potential biomarkers for SLE diagnosis. METHODS: Serum samples from 32 patients with SLE and 25 gender-matched healthy controls (HCs) were analysed by metabolic profiling based on liquid chromatography-tandem mass spectrometry metabolomics platform. The further validation for the potential biomarker was performed in an independent set consisting of 36 SLE patients and 30 HCs. RESULTS: The metabolite profiles of serum samples allowed differentiation of SLE patients from HCs. The levels of arachidonic acid, sphingomyelin (SM) 24:1, monoacylglycerol (MG) 17:0, lysophosphatidyl ethanolamine (lysoPE) 18:0, lysoPE 16:0, lysophosphatidyl choline (lysoPC) 20:0, lysoPC 18:0 and adenosine were significantly decreased in SLE patients, and the MG 20:2 and L-pyroglutamic acid were significantly increased in SLE group. In addition, L-pyroglutamic acid achieved an area under the receiver-operating characteristic curve of 0.955 with high sensitivity (97.22%) and specificity (83.33%) at the cut-off of 61.54 µM in the further targeted metabolism, indicating diagnostic potential. CONCLUSION: Serum metabolic profiling is differential between SLE patients and HCs and depicts increased L-pyroglutamic acid as a promising bitformatomarker for SLE.

Phyllotaxis: from classical knowledge to molecular genetics.

Plant organs are repetitively generated at the shoot apical meristem (SAM) in recognizable patterns. This phenomenon, known as phyllotaxis, has long fascinated scientists from different disciplines. While we have an enriched body of knowledge on phyllotactic patterns, parameters, and transitions, only in the past 20 years, however, have we started to identify genes and elucidate genetic pathways that involved in phyllotaxis. In this review, I first summarize the classical knowledge of phyllotaxis from a morphological perspective. I then discuss recent advances in the regulation of phyllotaxis, from a molecular genetics perspective. I show that the morphological beauty of phyllotaxis we appreciate is the manifestation of many regulators, in addition to the critical role of auxin as a patterning signal, exerting their respective effects in a coordinated fashion either directly or indirectly in the SAM.

Increasing Quantum Correlations Based on Measurement-Induced Disturbance via a Swapping Procedure with Two-Qubit Mixed States.

In this paper, quantum correlation (QC) swapping for certain separable two-qubit mixed states is treated. A QC quantifier, measurement-induced disturbance (MID) (Luo in Phys Rev A 77:022301, 2008), is employed to characterize and quantify QCs in the relevant states. Properties of all QCs in the swapping process are revealed. Particularly, it is found that MID can be increased through QC swapping for certain separable two-qubit mixed states.

Effects of miR-34c-5p on Sodium, Potassium, and Calcium Channel Currents in C2C12 Myotubes.

The aim of this study was to investigate the effects of miR-34c-5p on the main voltage-dependent ion channels in skeletal muscle cells. This study focused on the effects of miR-34c-5p on sodium, potassium, and calcium currents in C2C12 myoblasts. The miR-34c-5p overexpression group, knockdown group, and control group were differentiated for 7 days, fused into myotubes, and used for the whole-cell patch clamp recording. Compared with the control group, the whole-cell sodium current density of the other two groups had no significant changes. In the knockdown group, the delayed rectifier potassium current density was increased (statistically significant), and the whole-cell calcium channel current density did not change. In the overexpression group, the change of rectifier potassium current density was not obvious, while the peak calcium channel current density increased (- 9.23 ± 0.95 pA/pF, n = 6 cells for the overexpression group vs. - 6.48 ± 0.64 pA/pF, n = 7 cells for the control; p < 0.05). Changes in the expression of miR-34c-5p can affect the electrophysiological characteristics of calcium and potassium voltage-gated channels in C2C12 myotubes. Overexpression of miR-34c-5p increased whole-cell L-type calcium channel current (ICa,L), while miR-34c-5p knockdown increased whole-cell delayed rectifier potassium current (IKd).

Acute and Subacute Toxicity Study of Graphene-Based Tumor Cell Nucleus-Targeting Fluorescent Nanoprobes.

Graphene-based tumor cell nuclear targeting fluorescent nanoprobes (GTTNs) were synthesized in our laboratory as a kind of nanomaterial and showed good performance for both in vivo and in vitro imaging. GTTNs directly cross the cell membrane and specifically target the tumor cell nucleus via a cell membrane permeability targeting (CMPT) mechanism, which takes advantage of the increased permeability of the tumor cell membranes. GTTNs with a CMPT mechanism achieve high targeting efficiency in tumor tissues. With the tumor cell nucleus-targeting characterization, the GTTN distinguishes tumor cells at the single-cell level and recognizes the tumor tissue interface in a very early stage and shows great potential in clinical applications. Toxicity studies are extremely critical for clinical applications. Therefore, we studied the acute and subacute toxicity of GTTNs using an in vivo method and examined the following experimental indicators: mouse body weight, organ coefficients, serum biochemical parameters, and histological changes. The results showed that there were no significant differences in any indicators between the experimental and control mice. We also used an in vitro method to study the cytotoxicity of GTTNs in GES-1 (gastric epithelial cell) cells. Surprisingly, the results demonstrated over 80% cell viability when the incubation time reached up to 72 h under a 200 mg/L concentration of GTTNs, which indicated that GTTNs had low cytotoxicity. GTTNs barely showed any acute or subacute toxicity or cytotoxicity in vivo and in vitro, respectively, which supports their use for clinical applications.

Analytic Expression of Quantum Discords in Werner States under LQCC.

In this paper, quantum discords in a special kind of states, i.e., Werner states by local quantum operations and classical communication (LQCC) protocols (WLQCC states), are studied. Nineteen parameters to quantify the quantum discords are reduced to four parameters in terms of properties of Werner states and quantum discord. In the case of orthogonal projective measures, analytic expression of quantum discords in WLQCC states is analytically worked out. Some properties of the quantum discord in the WLQCC states are obtained, especially the variation relations between the quantum discords and the parameters characterizing the WLQCC states. By virtue of numerical computations, quantum discords in a Werner state before and after LQCC protocols are compared. It is found that quantum discord in any WLQCC state cannot exceed that in the original Werner state.

Morphogenesis of flattened unifacial leaves in Juncus prismatocarpus (Juncaceae).

To reveal the mode of morphogenesis of flattened unifacial leaves, we analysed the cell division direction and distribution on the leaf blade of Juncus prismatocarpus. Using the pulse-chase 5-ethynyl-2'-deoxyuridine method, we quantified and mapped the cell division direction on the leaf blade of J. prismatocarpus and compared the distribution of thickening cell divisions with the expression pattern of DROOPING LEAF (DL), a key gene involved in leaf blade thickening. Thickening cell divisions were the most abundant (> 45%) among all cell division directions on the leaf blade of J. prismatocarpus from the early plastochron 2 stage through the plastochron 3 stage. Mapping of cell divisions indicated that cell divisions in a particular direction were not restricted to a particular domain but were distributed diffusely throughout the entire cross-sectional area of the leaf blade. Gradient analysis indicated that the distribution of thickening cell divisions of the adaxial domain was denser than that of the abaxial domain. Contrary to the prolonged and diffuse distribution of thickening cell divisions, DL expression was transient and restricted in a narrow band. Our results suggest that a diffuse 'thickening meristem' plays the key role in the development of flattened unifacial leaves.

Size, Shape, and Protein Corona Determine Cellular Uptake and Removal Mechanisms of Gold Nanoparticles.

Size, shape, and protein corona play a key role in cellular uptake and removal mechanisms of gold nanoparticles (Au NPs). The 15 nm nanoparticles (NP1), the 45 nm nanoparticles (NP2), and the rod-shaped nanoparticles (NR) enter into cells via a receptor-mediated endocytosis (RME) pathway. The star-shaped nanoparticles (NS) adopt not only clathrin-mediated, but also caveolin-mediated endocytosis pathways. However, the 80 nm nanoparitcles (NP3) mainly enter into the cells by macropinocytosis pathway due to the big size. Furthermore, the results indicate that the presence of protein corona can change the uptake mechanisms of Au NPs. The endocytosis pathway of NP1, NP2, and NS changes from RME to macropinocytosis pathway and NR changes from RME to clathrin and caveolin-independent pathway under the non-fetal bovine serun (FBS)-coated condition. Both FBS-coated and non-FBS-coated of five types of Au NPs are released out through the lysosomal exocytosis pathway. The size, shape, and protein corona have an effect on the exocytosis ratio and amount, but do not change the exocytosis mechanism. The systematic study of the endocytosis and exocytosis mechanism of Au NPs with different sizes and shapes will benefit the toxicology evaluation and nanomedicine application of Au NPs.

Quantifying circular RNA expression from RNA-seq data using model-based framework.

MOTIVATION: Circular RNAs (circRNAs) are a class of non-coding RNAs that are widely expressed in various cell lines and tissues of many organisms. Although the exact function of many circRNAs is largely unknown, the cell type-and tissue-specific circRNA expression has implicated their crucial functions in many biological processes. Hence, the quantification of circRNA expression from high-throughput RNA-seq data is becoming important to ascertain. Although many model-based methods have been developed to quantify linear RNA expression from RNA-seq data, these methods are not applicable to circRNA quantification. RESULTS: Here, we proposed a novel strategy that transforms circular transcripts to pseudo-linear transcripts and estimates the expression values of both circular and linear transcripts using an existing model-based algorithm, Sailfish. The new strategy can accurately estimate transcript expression of both linear and circular transcripts from RNA-seq data. Several factors, such as gene length, amount of expression and the ratio of circular to linear transcripts, had impacts on quantification performance of circular transcripts. In comparison to count-based tools, the new computational framework had superior performance in estimating the amount of circRNA expression from both simulated and real ribosomal RNA-depleted (rRNA-depleted) RNA-seq datasets. On the other hand, the consideration of circular transcripts in expression quantification from rRNA-depleted RNA-seq data showed substantial increased accuracy of linear transcript expression. Our proposed strategy was implemented in a program named Sailfish-cir. AVAILABILITY AND IMPLEMENTATION: Sailfish-cir is freely available at https://github.com/zerodel/Sailfish-cir . CONTACT: tongz@medicine.nevada.edu or wanjun.gu@gmail.com. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Morphology effect on photocatalytic activity in Bi3Fe0.5Nb1.5O9.

In this work, the Aurivillius-phase ferroelectric Bi3Fe0.5Nb1.5O9 were synthesized by hydrothermal (BFNO-H) and solid state methods (BFNO-S), respectively. The BFNO-H shows a hierarchical morphology, which is stacked by intersecting single-crystal nanosheets with {001} and {110} exposed facets, while the BFNO-S shows disorganized micron-scale morphology. BFNO-H shows a much stronger photodegradation activity (10.4 times and 9.8 times) than BFNO-S in the visible-light photodegradation of rhodamine B (RhB) and salicylic acid. The higher photodegradation activity of BFNO-H was firstly ascribed to the hierarchical structure and the larger specific surface area (16.586 m2 g-1) because a large specific surface area can increase reactive sites and shorten photogenerated carrier migration distance. However, after being normalized by the specific surface area, BFNO-H still performs better than BFNO-S, implying that the specific surface area is not the only factor that determines the photocatalytic activity. Considering that the built-in electric field originating from spontaneous polarization in Bi3Fe0.5Nb1.5O9 has existed in both ab plane and c direction, it matches well with the {001} and {110} exposed facets of BFNO-H nanosheets. This appropriate matching in BFNO-H nanosheets may improve the separation and transmission of photogenerated electron-hole pairs and further enhance its photocatalytic activity. Moreover, the trapping experiments reveals that holes (h +) are the main active species and hole-derived oxidation is the main redox reaction during photodegradation of organic pollutions.

Serum Levels of TRIM72 Are Lower among Patients with Colon Cancer: Identification of a Potential Diagnostic Marker.

Colon cancer is one of the most common malignancies causing the majority of cancer-related deaths worldwide. The tripartite motif family protein 72 (TRIM72), also known as mitsugumin 53, acts as an E3 ubiquitin ligase. TRIM72 is involved in insulin resistance and metabolic syndrome, which are risk factors of colon cancer. However, the correlation between TRIM72 and colon cancer remains unknown. In the present study, we explored the expression profile of TRIM72 in colon cancer tissues and the diagnostic value of serum TRIM72 in colon cancer. The receiver operating characteristic (ROC) curves were applied for evaluating the diagnostic value of serum TRIM72. We thus found that immunoreactive TRIM72 levels were significantly lower in colon cancer tissues than those in normal colon tissues. Moreover, serum TRIM72 levels were significantly lower in colon cancer patients than those in healthy volunteers. Importantly, the lower serum TRIM72 levels were associated with advanced clinical stage, lymph node, and distant metastases in colon cancer patients. The ROC curve analysis showed that serum TRIM72 has a superior diagnostic value (the area under the curve (AUC) = 0.829) than the traditional tumor biomarkers, carcinoembryonic antigen (CEA) (AUC = 0.707) and carbohydrate antigen 19-9 (CA199) (AUC = 0.750), and the combination of TRIM72 with CEA and CA199 showed the best diagnostic value for colon cancer (AUC = 0.928). In conclusion, serum TRIM72 may be a potential biomarker for the diagnosis and the prognosis of colon cancer.

Anisotomous dichotomy results from an unequal bifurcation of the original shoot apical meristem in Diphasiastrum digitatum (Lycopodiaceae).

PREMISE OF THE STUDY: Two types of dichotomy are recognized in Lycopodiaceae: isotomous (equal) and anisotomous (unequal). Anisotomous dichotomy (anisotomy) has been hypothesized to result from unequal growth of an equal bifurcation of the original shoot apical meristem (SAM). Diphasiastrum digitatum (Lycopodiaceae) exhibits anisotomy at various locations. We thus used D. digitatum to test this classic hypothesis about anisotomy. METHODS: Transverse areas of original and derived SAMs of anisotomy exhibited by the rhizome and the vertical aerial vegetative stem were measured using scanning electron microscopy. The difference between half of the original SAM and one derived SAM in terms of transverse area were compared using paired t-tests. KEY RESULTS: During the anisotomy exhibited by the rhizome SAM, 77.4% of the transverse area of the original rhizome SAM contributed to the derived rhizome SAM. During the first anisotomy exhibited by the vertical aerial vegetative stem SAM, 66.2% of the transverse area of the original vertical aerial vegetative stem SAM contributed to the derived vertical aerial vegetative stem SAM. During the second anisotomy exhibited by the vertical aerial vegetative stem SAM, 49.4% of the transverse area of the original vertical aerial vegetative stem SAM contributed to the derived vertical aerial vegetative stem SAM. Nonetheless, the shape of the two derived SAMs differed though they did not differ in size. CONCLUSIONS: In D. digitatum, anisotomy results from an unequal bifurcation of the original SAM. This finding sheds light on plant body architecture evolution as well as plant organ (megaphyllous leaf) evolution.