Ultrastructure of the amoeboid flagellate <Emphasis Type="Italic">Thaumatomonas zhukovi</Emphasis> Mylnikov et Mylnikov (Thaumatomonadida (Shirkina) Karpov, 1990)

The ultrastructure of the amoeboid flagellate Thaumatomonas zhukovi sp. is presented. The cell is covered by cell body scales that formed on the surface of mitochondria. Capturing bacteria, the pseudopodia emerge from the ventral groove, which is supported by two longitudinal microtubular bands. The heterodynamic flagella emerge from the small flagellar pocket. Both flagella are covered by cone-shaped scales and thin twisted mastigonemes. The transitional zone of the flagella contains a thin-walled cylinder. The transversal plate of the flagella rises above the cell surface. The kinetosomes lie parallel to each other. The flagellar root system consists of three microtubular bands and a fibrillar rhizoplast. The vesicular nucleus and the Golgi apparatus have typical structures. The cytoplasm contains microbodies and food vacuoles. Mitochondria contain tubular cristae. Extrusive organelles (kinetocysts), which contain amorphous material and a capsule, were found in the cytoplasm. The capsule consists of a theca and a cylinder. The resemblance of Thaumatomonas zhukovi to other thaumatomonads is discussed.     

The ultrathin structure of amoeboid flagellate <Emphasis Type="Italic">Thaumatomastix</Emphasis> sp. (Thaumatomonadida (Shirkina) Karpov, 1990)

The ultrathin structure of amoeboid flagellate Thaumatomastix sp. is considered. The cell is surrounded by two-layered triangular scales. They are formed on the surface of mitochondria. Pseudopodia grabbing bacteria run from ventricular furrow, which is armored with two longitudinal bands of microtubules. Heterodynamic flagella run from small flagellar pocket. Long back flagellum has thin mastigonemes. Proximal area of short flagellum is covered with flat oval scales. Transitional flagellant zone has no spiral or other additional elements. Transverse plate is localized above cell surface. Kinetosomes are parallel to each other. Vesicular nucleus and Golgi apparatus have typical structure. Oval mitochondria contain tubular cristae. Within cytoplasm, extrusive organelles (kinetocysts) containing amorphous material and capsule were found. The latter consists of muff and cylinder. Plasmodial and cystic phases of development have not been discovered. Contractile vacuole is absent. The resemblance between Thaumatomastix sp. and other thaumatomonads has been discussed.     

Inverse relationship of Ca<Superscript>2+</Superscript>-dependent flagellar response between animal sperm and prasinophyte algae

Symmetry/asymmetry conversion of eukaryotic flagellar waveform is caused by the changes in intracellular Ca²⁺. Animal sperm flagella show symmetric or asymmetric waveform at lower or higher concentration of intracellular Ca²⁺, respectively. In Chlamydomonas, high Ca²⁺ induces conversion of flagellar waveform from asymmetric to symmetry, resulting in the backward movement. This mirror image relationship between animal sperm and Chlamydomonas could be explained by the distinct calcium sensors used to regulate the outer arm dyneins (Inaba 2015). Here we analyze the flagellar Ca²⁺-response of the prasinophyte Pterosperma cristatum, which shows backward movement by undulating four flagella, the appearance similar to animal sperm. The moving path of Pterosperma shows relatively straight in artificial seawater (ASW) or ASW in the presence of a Ca²⁺ ionophore A23187, whereas it becomes circular in a low Ca²⁺ solution. Analysis of flagellar waveform reveals symmetric or asymmetric waveform propagation in ASW or a low Ca²⁺ solution, respectively. These patterns of flagellar responses are completely opposite to those in sperm flagella of the sea urchin Anthocidaris crassispina, supporting the idea previously proposed that the difference in flagellar response to Ca²⁺ attributes to the evolutional innovation of calcium sensors of outer arm dynein in opisthokont or bikont lineage.     

<Emphasis Type="Italic">Chlamydomonas fla</Emphasis> mutants reveal a link between deflagellation and intraflagellar transport

Background

Cilia and flagella are often lost in anticipation of mitosis or in response to stress. There are two ways that a cell can lose its flagella: resorption or deflagellation. Deflagellation involves active severing of the axoneme at the base of the flagellum; this process is defective in Chlamydomonas fa mutants. In contrast, resorption has been thought to occur as a consequence of constitutive disassembly at the tip in the absence of continued assembly, which requires intraflagellar transport (IFT). Chlamydomonas fla mutants are unable to build and maintain flagella due to defects in IFT.

Results

fla10 cells, which are defective in kinesin-II, the anterograde IFT motor, resorb their flagella at the restrictive temperature (33°C), as previously reported. We find that in standard media containing ~300 microM calcium, fla10 cells lose flagella by deflagellation at 33°C. This temperature-induced deflagellation of a fla mutant is not predicted by the IFT-based model for flagellar length control. Other fla mutants behave similarly, losing their flagella by deflagellation instead of resorption, if adequate calcium is available. These data suggest a new model whereby flagellar resorption involves active disassembly at the base of the flagellum via a mechanism with components in common with the severing machinery of deflagellation. As predicted by this model, we discovered that deflagellation stimuli induce resorption if deflagellation is blocked either by mutation in a FA gene or by lack of calcium. Further support for this model comes from our discovery that fla10-fa double mutants resorb their flagella more slowly than fla10 mutants.

Conclusions

Deflagellation of the fla10 mutant at the restrictive temperature is indicative of an active disassembly signal, which can manifest as either resorption or deflagellation. We propose that when IFT is halted by either an inactivating mutation or a cellular signal, active flagellar disassembly is initiated. This active disassembly is distinct from the constitutive disassembly which plays a role in flagellar length control.

     

Ultrastructure of Sperm in <Emphasis Type="Italic">Mercenaria stimpsoni</Emphasis> and <Emphasis Type="Italic">Mactra chinensis</Emphasis> (Mollusca: Bivalvia) from the Sea of Japan

The ultrastructure of the sperm of the common bivalve species Mercenaria stimpsoni and Mactra chinensis from Peter the Great Bay is described. The sperm structure is typical for animals with external insemination. The sperm consists of a head, middle part, and flagellum. The sperm head of M. stimpsoni has a curved crescent form and includes the nucleus and acrosome; the head length is 9.8 μm. The acrosome is subdivided to the acrosome granule and the periacrosomal material. There are 4 mitochondria of about 0.8 μm in size in the middle part of the spermatozoon. The mitochondria surround the centriolar apparatus, which consists of proximal and distal centrioles located at a right angle. The axoneme originates from the distal centriole. The sperm of M. chinensis is barrel-shaped, with a head length of 3.2 μm. The acrosome is relatively larger, and its height is 1–1.2 μm. There are also 4 mitochondria 0.6–0.8 μm in the middle part of the spermatozoon. The sperm structure of the described species is typical of the families to which the mollusks belong, with insignificant variations.     

On the multicomponent nature of <Emphasis Type="Italic">Halobacterium salinarum</Emphasis> flagella

Filaments of the flagellum of the halophilic archaeon Halobacterium salinarum consist of five flagellins: A1, A2, B1, B2, and B3, which are encoded by five genes localized in tandem in two flgA and flgB operons. While the role of flagellins A1 and A2 has been determined, the role of the proteins, B operon products, is still unclear. A mutant strain of H. salinarum with deleted A and B flagellin genes (ΔflgAΔflgB) has been obtained for the first time. This strain has been used to create and analyze the strains carrying only individual B1 or B3 flagellin genes. Cells of the ΔflgAΔflgB strain were shown to have short filamentous formations, 7–8 nm thick, which we have named as X-filaments. It has been shown that X-filaments consist of a protein immunologically related to flagellins A and B. Expression of the B1 and B3 genes is suppressed in the absence of A1, A2, and B2. It has been shown that flagellins B1 and B3 cannot be substituted for flagellin B2 upon the formation of a curved hook-like structure, which serves as a connecting element between the flagellar filament and the motor axis. The multicomponent nature of flagella is discussed in the light of their possible involvement in other cell processes besides providing motility.     

Effect of gut bacteria on fitness of the Chinese citrus fly, <Emphasis Type="Italic">Bactrocera minax</Emphasis> (Diptera: Tephritidae)

A likely symbiotic association between tephritid fruit flies and gut bacteria has been recognized since the beginning of the last century. However, direct evidence for a link between gut bacteria and fruit fly fitness is still limited or absent for many species. Similar to other tephritids, the gut of Bactrocera minax (Enderlein) (Diptera: Tephritidae) is known to contain bacteria throughout the life stage, but what, if any, impact these bacteria have on B. minax fitness is entirely unknown. In order to elucidate the effects of bacteria on the fitness of B. minax, resident bacteria were isolated from the adult gut using culture-dependent techniques. Adult fly diets were subsequently supplemented with three bacterial isolates (Klebsiella pneumonia, Citrobacter braakii and Pantoea dispersa), or bacteria were removed from flies by antibiotics treatment: untreated adults provided a control. Adult fitness parameters (male and female longevity, female fecundity, male copulation number) were measured for the two treatments and one control group. Results were complex depending on the fitness parameter measured and the bacterial species. Compared to the controls, antibiotic treated B. minax had significantly decreased fecundity, but male and female longevity was increased. When flies were fed diets supplemented with any of the three bacterial isolates, female fecundity was significantly enhanced. However, only Citrobacter braakii significantly increased male mating frequency than control males. The results show that gut bacteria directly influence fitness of B. minax, but impacts are dependent on the bacterial species and the fitness parameters measured.     

The histology of the scales ofOrvikuina (Palaeoniscoidea)

The analysis byØrvig of the structure of the scales ofScanilepis andOrvikuina is discussed. It is shown that the analysis, because it ignores the physical size of the scales, incorrectly leads to the conclusion that the scales ofOrvikuina show a stage of specialisation towards the higher teleostean condition further than, but of the same kind as, that shown byScanilepis. The specialisation ofOrvikuina is in fact of a different kind from that ofScanilepis.     

Chromosomal <Emphasis Type="Italic">flhB1</Emphasis> gene of the alphaproteobacterium <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> Sp245 is essential for correct assembly of both constitutive polar flagellum and inducible lateral flagella

Azospirillum brasilense has the ability of swimming and swarming motility owing to the work of a constitutive polar flagellum and inducible lateral flagella, respectively. The interplay between these flagellar systems is poorly understood. One of the key elements of the flagellar export apparatus is the protein FlhB. Two predicted flhB genes are present in the genome of A. brasilense Sp245 (accession nos. HE577327–HE577333). Experimental evidence obtained here indicates that the chromosomal coding sequence (CDS) AZOBR_150177 (flhB1) of Sp245 is essential for the production of both types of flagella. In an flhB1::?Omegon-Km mutant, Sp245.1063, defects in polar and lateral flagellar assembly and motility were complemented by expressing the wild-type flhB1 gene from plasmid pRK415. It was found that Sp245.1063 lost the capacity for slight but statistically significant decrease in mean cell length in response to transfer from solid to liquid media, and vice versa; in the complemented mutant, this capacity was restored. It was also shown that after the acquisition of the pRK415-harbored downstream CDS AZOBR_150176, cells of Sp245 and Sp245.1063 ceased to elongate on solid media. These initial data suggest that the AZOBR_150176-encoded putative multisensory hybrid sensor histidine kinase–response regulator, in concert with FlhB1, plays a role in morphological response of azospirilla to changes in the hardness of a milieu.     

Biochemical evidence bearing on the domestication of<Emphasis Type="Italic">Phaseolus</Emphasis> (Fabaceae) beans

The genusPhaseolus (Fabaceae) consists of some 50 species, all of which are distributed in the Americas. Four of these contain cultigens.P. vulgaris (common bean),P. lunatus (lima bean),P. acutifolius (tepary bean),P. coccineus subsp.coccineus (runner bean); andP. coccineus subsp.polyanthus (no English vernacular name). Biochemical markers—phaseolin seed storage protein and isozymes—have provided new evidence on the organization of the first three species. Domestication has possibly caused a strong reduction in genetic diversity inP. vulgaris andP. acutifolius. BothP. vulgaris andP. lunatus cultivars result from at least two independent domestications, in Mesoamerica and in the Andes. These two species consist of two gene pools, each of which includes wild ancestors and their respective cultivated descendants. Our findings suggest the need for additional emphasis on genetic conservation of wild ancestors and their use in breeding programs and for a comparison of inter-gene pool vs. intra-gene pool crosses in breeding programs.     

Metatrancriptomic analysis from the Hepatopancreas of adult white leg shrimp (<Emphasis Type="Italic">Litopenaeus vannamei)</Emphasis>

The amoeba, Mayorella viridis contains several hundred symbiotic green algae in its cytoplasm. Transmission electron microscopy revealed strong resemblance between symbiotic algae from M. viridis the symbiotic Chlorella sp. in the perialgal vacuoles of Paramecium bursaria and other ciliates. Although it is thought that the M. viridis and symbiotic algae could be model organisms for studying endosymbiosis between protists and green algae, few cell biological observations of the endosymbiosis between M. viridis and their symbiotic algae have been published. In this study, we characterized the specificity of endosymbiotic relationships between green algae and their hosts. Initially, we established stable cultures of M. viridis in KCM medium by feeding with Chlorogonium capillatum. Microscopic analyses showed that chloroplasts of symbiotic algae in M. viridis occupy approximately half of the algal cells, whereas those in P. bursaria occupy entire algal cells. The symbiotic algae in P. bursaria contain several small spherical vacuoles. The labeling of actin filaments using Acti-stain? 488 Fluorescent Phalloidin revealed no relationship between host actin filaments and symbiotic algal localization, although the host mitochondria were localized around symbiotic algae. Symbiotic algae from M. viridis could infect algae-free P. bursaria but could not support P. bursaria growth without feeding, whereas the original symbiotic algae of P. bursaria supported its growth without feeding. These data indicated the specificity of endosymbiotic algae relationships in M. viridis and P. bursaria.     

Microstructural characterization of the body key scale morphology in six Iranian endemic <Emphasis Type="Italic">Aphanius</Emphasis> species (Cyprinodontidae): Their taxonomic and evolutionary significance

The microstructural characteristics of the body key scales were described for six Iranian endemic Aphanius Nardo, 1827 species. Among them, five species genetically belong to the “Iranian inland and inland-related Aphanius species, IIRAS” group, while A. ginaonis (Holly, 1929) belongs to the “brackish water species” group. General scale shape in the studied Aphanius species was cycloid with the exception of A. ginaonis, which has cycloid scales with few spinous-like structures present in the posterior edge of the scales. Considering phylogenetic relationships of the studied taxa, the most likely explanation for presence of the spinous scales in Aphanius species is the primary existence of these scales in this group. In addition, among the IIRAS group, the most different lepidont was recognized in scales of the A. isfahanensis Hrbek, Keivany and Coad, 2006 (pointed), which is similar to lepidonts in the A. ginaonis. The scales of these species are polygonal with similar typology. The similarity in lepidont morphology between A. ginaonis and A. isfahanensis can explained by (i) the primary existence of the rounded lepidont in this group, and (ii) since A. ginaonis and A. isfahanensis belong to two separate phylogenetic lineages, therefore, similarity in their lepidont morphology could be results of convergent evolution.     

Root-associated bacteria influencing mycelial growth of <Emphasis Type="Italic">Tricholoma matsutake</Emphasis> (pine mushroom)

Tricholoma matsutake is an ectomycorrhizal fungus usually associated with Pinus densiflora in South Korea. Fruiting bodies (mushrooms) of T. matsutake are economically important due to their attractive aroma; yet, T. matsutake is uncultivatable and its habitat is rapidly being eradicated due to global climate change. Root-associated bacteria can influence the growth of ectomycorrhizal fungi that co-exist in the host rhizosphere and distinctive bacterial communities are associated with T. matsutake. In this study, we investigated how these bacterial communities affect T. matsutake growth by isolating bacteria from the roots of P. densiflora colonized by ectomycorrhizae of T. matsutake and co-culturing rootassociated bacteria with T. matsutake isolates. Thirteen species of bacteria (27 isolates) were found in pine roots, all belonging to the orders Bacillales or Burkholderiales. Two species in the genus Paenibacillus promoted the growth of T. matsutake in glucose poor conditions, likely using soluble metabolites. In contrast, other bacteria suppressed the growth of T. matsutake using both soluble and volatile metabolites. Antifungal activity was more frequent in glucose poor conditions. In general, pine rhizospheres harbored many bacteria that had a negative impact on T. matsutake growth and the few Paenibacillus species that promoted T. matsutake growth. Paenibacillus species, therefore, may represent a promising resource toward successful cultivation of T. matsutake.     

Aseptic rearing procedure for the stinkbug <Emphasis Type="Italic">Plautia stali</Emphasis> (Hemiptera: Pentatomidae) by sterilizing food-derived bacterial contaminants

The stinkbug Plautia stali Scott is a notorious agricultural pest whose posterior midgut hosts specific bacteria essential for its growth and survival, highlighted as an experimental model for symbiosis studies. Some symbiotic bacteria of P. stali are cultivable, found free-living in and acquired from the environment, and, furthermore, some free-living environmental bacteria are potentially capable of establishing symbiotic association with P. stali. In this context, it is expected that such environmental bacteria may occasionally contaminate and infect the experimental insects maintained in the laboratory, which could potentially affect the functional analyses of the symbiosis. Here we report that such contamination events do occur under a laboratory rearing conditions for P. stali. When symbiont-deprived newborn nymphs from surface-sterilized eggs were reared in sterilized plastic containers with autoclaved water, most of them died as nymphs presumably as a result of aposymbiosis, but only a small fraction could attain adulthood and the adult insects were all infected with γ-proteobacteria allied to Pantoea and Enterobacter. A variety of bacteria, mainly Bacillus and also Pantoea and Enterobacter, were detected from peanuts and soybeans provided as food for P. stali. Autoclaving of peanuts and soybeans eradicated these bacteria but negatively affected the host survival, whereas ethanol sterilization of peanuts and soybeans removed Pantoea and Enterobacter, but not Bacillus, without negative effects on the host survival. On the basis of these results, we established a practical procedure for aseptic rearing of P. stali, which will enable reliable and strict analyses of host–symbiont interactions in the model symbiotic system.     

Reconstitution of <Emphasis Type="Italic">Salmonella</Emphasis> Flagella attached to Cell Bodies

THE reconstitution in vitro of flagellar filaments from their component flagellin monomers in Salmonella has shown that the filaments have structural polarity and grow at an end distal to the cell body¹; flagella in vivo also grow from their tips^2,3. This suggests that even when flagella are attached to living cells, filaments may be reconstituted from exogenous flagellin monomers at the tips in appropriate conditions. In spite of some negative results⁴, we have been encouraged^5–10 to re-examine the question.     

Antimicrobial activity of endogenous peptides of the moss <Emphasis Type="Italic">Physcomitrella patens</Emphasis>

Plant and animal cells contain pools of endogenous peptides, which are the degradation products of functionally active proteins. It is known that these peptides can possess biological activity; however, the functions of most of them are unknown. The goal of the present study was to estimate the antimicrobial potential of endogenous peptides resulting from the degradation of functional proteins in cells of the moss Physcomitrella patens. Earlier, 117 peptides possessing an antimicrobial potential predicted in silico have been identified in the peptidomes of three types of P. patens cells by mass spectrometry. In the present work, the antimicrobial activity of six of these peptides toward the gram-positive bacteria Bacillus subtilis SHgw and Clavibacter michiganensis pv. michiganensis and gram-negative bacteria Escherichia coli K12 and Xanthomonas arboricola 3004 has been revealed. The results have shown that three of six peptides inhibit the growth of the phytopathogenic bacteria X. arboricola and C. m. pv. michiganensis; four peptides inhibit the growth of the gram-negative bacterium E. coli K12, and one peptide inhibits the growth of the gram-positive bacterium B. subtilis. It has been found that the peptides inhibiting the bacterial growth are predominantly the fragments of ribosomal proteins. The work confirms the potential of the biological activity of peptides that are the degradation products of functional proteins.     

Nectar and oleiferous trichomes as floral attractants in <Emphasis Type="Italic">Bulbophyllum saltatorium</Emphasis> Lindl. (Orchidaceae)

Although many Orchidaceae have deceit flowers that produce no reward, the most common reward, when present, is nectar. Bulbophyllum, however, is unusual in that the labellar secretions of most species investigated to date lack sugars, and, therefore, cannot be considered true nectar. The African species Bulbophyllum saltatorium is an exception in that it produces not only nectar but also possesses specialized, capitate oleiferous trichomes. The nectary of B. saltatorium is borne on the labellum and is represented by a deep, narrow, median longitudinal groove, having a small aperture, and flanked by trichomes. Isodiametric epidermal cells lining this groove secrete nectar which collects both in the groove and on the surface of the labellum. As well as a nectary, the labellum of B. saltatorium also bears three types of unicellular trichomes: the longest trichomes are borne distally and abaxially; the marginal ones form a rim around the entire labellum, and finally, massive, capitate trichomes occur proximally and adaxially. These are oleiferous, containing large quantities of oil which might function as precursors of volatile components of fragrance or provide a food-reward. To the best of our knowledge, this is the first time for such oleiferous trichomes to be described for Bulbophyllum. Therefore, apart from their color and markings, flowers of this species are able to attract pollinators in at least two, possibly three ways: food-reward in the form of nectar; fragrance; and possibly food-rewards in the form of food-hairs.     

First Record of Larval Secretions of <Emphasis Type="Italic">Cochliomyia macellaria</Emphasis> (Fabricius, 1775) (Diptera: Calliphoridae) Inhibiting the Growth of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

Maggot debridement therapy (MDT) consists on the intentional and controlled application of sterilized larvae of the order Diptera on necrotic skin lesions with the purpose of cleaning necrotic tissue and removing pathogenic bacteria. During MDT, a marked antimicrobial activity has been reported in literature specially associated with antibacterial substances from Lucilia sericata (Meigen); however, regarding Cochliomyia macellaria (Fabricius), little is known. This study aimed to evaluate in vitro inhibition of bacterial growth of Pseudomonas aeruginosa and Staphylococcus aureus in contact with excretions and secretions (ES) from C. macellaria larvae. Larval ES were extracted in sterile distilled water and divided in three groups: ES, containing 400 μL of autoclaved ES; ES+BAC, containing 400 μL of autoclaved ES+0.5-μL bacterial inoculum; and CONT-BAC, containing 400 μL of sterile distilled water +0.5 μL of bacterial inoculum. Aliquots of each experimental group were plated by spreading onto Petri dishes. Seedings were made at 0, 1, 2, 4, and 12 h after the extraction of ES. In ES+BAC groups, inhibition of S. aureus was verified between times 1 and 2 h and P. aeruginosa was inhibited between 0 and 4 h. There was no growth observed in any ES group. In the CONT-BAC groups, the number of colonies from time 4 h became countless for S. aureus and decreased for P. aeruginosa. As reported in the literature, we note here that ES have excellent bactericidal activity for both gram-positive and gram-negative bacteria, and this study shows for the first time the action of the bactericidal activity of exosecretions of C. macellaria against S. aureus and P. aeruginosa.     

The lateral line system and its innervation in <Emphasis Type="Italic">Lateolabrax japonicus</Emphasis> (Percoidei <Emphasis Type="Italic">incertae sedis</Emphasis>) and two apogonids (Apogonidae), with special reference to superficial neuromasts (Teleostei: Percomorpha)

The lateral line system and its innervation were examined in a generalized perch-like species, Lateolabrax japonicus (Percoidei incertae sedis), and compared with those in two species of Apogonidae (Fowleria variegata in Apogonichthyini and Ostorhinchus doederleini in Ostorhinchini) characterized by proliferated superficial neuromasts (SNs) on the head, trunk lateral line scales and caudal fin. The total number of SNs differed greatly between the two groups, being 271 in the former, and 2,403 and 4,088 in the latter. The mandibular ramus (MDR) was extensively ramified in the head of the apogonids, with three additional branches that were absent in L. japonicus, innervating 1,117 SNs in F. variegata and 1,928 in O. doederleini. In the apogonids, the additional anterodorsal branch of the MDR coursed parallel to the buccal ramus anteriorly (on the interorbital space) and to the supratemporal ramus posteriorly (on the temporal region). The two parallel portions supplied numerous SN rows forming a characteristic crosshatch pattern, the branch and two rami distributing to transverse and longitudinal rows, respectively. In the two groups, the trunk lateral line scales each housed a canal neuromast (CN; partly replaced by an SN in F. variegata). In addition, one to four (in L. japonicus) and three to 55 (in the apogonids) SNs occurred on each lateral line scale, the pattern of SN innervation being identical in having two types of branches; one innervated a CN and SNs, and the other SN(s) only. The latter type extended only to a limited number of scales in L. japonicus, but to nearly all or all scales in the apogonids. Compared with F. variegata, branches of the respective types were more finely ramified with greater number of SNs in O. doederleini.