Phylogenetic analyses of <Emphasis Type="Italic">Uromyces viciae-fabae</Emphasis> and its varieties on <Emphasis Type="Italic">Vicia</Emphasis>, <Emphasis Type="Italic">Lathyrus</Emphasis>, and <Emphasis Type="Italic">Pisum</Emphasis> in Japan

A pea rust fungus, Uromyces viciae-fabae, has been classified into two varieties, var. viciae-fabae and var. orobi, based on differences in urediniospore wall thickness and putative host specificity in Japan. In principal component analyses, morphological features of urediniospores and teliospores of 94 rust specimens from Vicia, Lathyrus, and Pisum did not show definite host-specific morphological groups. In molecular analyses, 23 Uromyces specimens from Vicia, Lathyrus, and Pisum formed a single genetic clade based on D1/D2 and ITS regions. Four isolates of U. viciae-fabae from V. cracca and V. unijuga could infect and sporulate on P. sativum. These results suggest that U. viciae-fabae populations on different host plants are not biologically differentiated into groups that can be recognized as varieties.Contribution no. 184, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

The <Emphasis Type="Italic">Caenorhabditis</Emphasis><Emphasis Type="Italic">briggsae</Emphasis> genome contains active <Emphasis Type="Italic">CbmaT1</Emphasis> and <Emphasis Type="Italic">Tcb1</Emphasis> transposons

The maT clade of transposons is a group of transposable elements intermediate in sequence and predicted protein structure to mariner and Tc transposons, with a distribution thus far limited to a few invertebrate species. We present evidence, based on searches of publicly available databases, that the nematode Caenorhabditis briggsae has several maT-like transposons, which we have designated as CbmaT elements, dispersed throughout its genome. We also describe two additional transposon sequences that probably share their evolutionary history with the CbmaT transposons. One resembles a fold back variant of a CbmaT element, with long (380-bp) inverted terminal repeats (ITRs) that show a high degree (71%) of identity to CbmaT1. The other, which shares only the 26-bp ITR sequences with one of the CbmaT variants, is present in eight nearly identical copies, but does not have a transposase gene and may therefore be cross mobilised by a CbmaT transposase. Using PCR-based mobility assays, we show that CbmaT1 transposons are capable of excising from the C. briggsae genome. CbmaT1 excised approximately 500 times less frequently than Tcb1 in the reference strain AF16, but both CbmaT1 and Tcb1 excised at extremely high frequencies in the HK105 strain. The HK105 strain also exhibited a high frequency of spontaneous induction of unc-22 mutants, suggesting that it may be a mutator strain of C. briggsae.     

Morphological and phylogenetic analyses of <Emphasis Type="Italic">Uromyces appendiculatus</Emphasis> and <Emphasis Type="Italic">U. vignae</Emphasis> on legumes in Japan

Uromyces appendiculatus, inclusive of three varieties, is distinguished from U. vignae primarily by the position of urediniospore germ pores and putative host specificity. However, opinions concerning these morphological and physiological features as taxonomic characters have varied greatly, and distinction of these species has often been confused. To clarify the taxonomy of these two species, morphological features of urediniospores and teliospores of 225 rust fungus specimens on species of Phaseolus, Vigna, Apios, Lablab, and Dunbaria were examined by light microscopy and scanning electron microscopy. Forty-five specimens were subjected to molecular phylogenetic analyses. As a result, the position of germ pores in urediniospores and the teliospore-wall thickness were considered as good characters to separate three morphological groups. In molecular analyses, the specimens fell into two and three clades based on the nucleotide sequence at D1/D2 domain of LSU rDNA and ITS regions, respectively. One of the D1/D2 clades corresponded to one morphological group whereas another D1/D2 clade included two other morphological groups. In contrast, each of the three ITS clades corresponded to a separate morphological group. Neither morphological groups nor molecular clades were host limited. It is suggested that the three morphological groups that corresponded to three distinct ITS clades constitute distinct species.Contribution no. 186 from the Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

Revision of <Emphasis Type="Italic">Heterosavia</Emphasis>, stat. nov., with notes on <Emphasis Type="Italic">Gonatogyne</Emphasis> and <Emphasis Type="Italic">Savia</Emphasis> (Phyllanthaceae)

Heterosavia (Phyllanthaceae) is segregated from Savia (tribe Bridelieae), recognized at generic rank, and placed in tribe Phyllantheae. Floral, fruit, leaf anatomical, leaf venation, and pollen characters of the neotropical taxa previously united as Savia including Gonatogyne are discussed and illustrated. Keys to the three genera and to the species of Heterosavia are presented. Four species (all new combinations), Heterosavia bahamensis, H. erythroxyloides, H. laurifolia, and H. maculata, are recognized. The new combinations Heterosavia laurifolia var. intermedia and H. maculata var. clementis are proposed. The names Heterosavia, H. erythroxyloides, H. laurifolia, Savia clementis, S. clusiifolia, S. clusiifolia var. fallax, and S. longipes are lectotypified. Distribution maps and conservation assessments (IUCN ratings) of Heterosavia species and varieties are provided.     

New species and a new combination in the <Emphasis Type="Italic">Hyphopichia</Emphasis> and <Emphasis Type="Italic">Yarrowia</Emphasis> yeast clades

Three new species of Candida and a new combination in the genus Hyphopichia are proposed from phylogenetic analysis of nucleotide divergence in domains D1/D2 of the large subunit (26S) rDNA. The new taxa and their type strains are the following: Candida bentonensis sp. nov. (NRRL YB-2364, CBS 9994), Candida hispaniensis sp. nov. (NRRL Y-5580, CBS 9996), Candida pseudorhagii sp. nov. (NRRL YB-2076, CBS 9998) and Hyphopichia heimii comb. nov. (NRRL Y-7502, CBS 6139), basionym Pichia heimii Pignal. Phylogenetic analysis placed C. pseudorhagii and H. heimii in the Hyphopichia clade whereas C. bentonensis and C. hispaniensis are members of the Yarrowia clade.     

<Emphasis Type="Italic">SQUA</Emphasis>-like genes in the orchid <Emphasis Type="Italic">Phalaenopsis</Emphasis> are expressed in both vegetative and reproductive tissues

The SQUA family (AP1/FUL family) of MADS-box genes plays an important role in the transition from the vegetative to the reproductive development of angiosperms, and its origin might be concurrent with fixation of floral structure in angiosperms. Here, we isolated two Phalaenopsis MADS-box genes designated ORAP11 and ORAP13, both of which belong to the monocot FUL-like clade of the SQUA family. RT-PCR showed that both genes are strongly expressed in the floral bud, and also detected in the vegetative organs. During later stages, ORAP11 was only detected in the column, but ORAP13 signal was absent from all of the floral organs. In-situ hybridization experiments detected both genes in the tips and margins of developing petals and lips, the developing column, and ovule. Over-expression of both genes in tobacco induced early flowering and changed plant architecture. Our results suggest that in Phalaenopsis, both genes might share partly redundant activities and play important roles in the process of floral transition and morphological architecture. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Redescriptions of the Indo-Pacific atherinid fishes <Emphasis Type="Italic">Atherinomorus forskalii</Emphasis>, <Emphasis Type="Italic">Atherinomorus lacunosus</Emphasis>, and <Emphasis Type="Italic">Atherinomorus pinguis</Emphasis>

The Indo-Pacific marine atherinid fishes Atherinomorus forskalii (Rüppell, 1838), Atherinomorus lacunosus (Forster, 1801), and Atherinomorus pinguis (Lacepède, 1803) are redescribed as valid species based on the types and non-type specimens collected throughout the Indo-Pacific. They are similar to each other chiefly in having a wide midlateral band (almost the same or greater than the midlateral scale width), large mouth (posterior tip of upper jaw reaching to or beyond a vertical through anterior margin of pupil), and no distinct tubercle at the posterior end of the dentary. All three species are distinguishable from congeners by those characters. The three species have long been confused with each other or synonymized erroneously as a single species. Atherinomorus forskalii, known from the Red Sea and eastern Mediterranean, differs from Atherinomorus lacunosus and Atherinomorus pinguis in having conspicuous, large endopterygoid teeth, forming obvious tooth ridges. Atherinomorus lacunosus, widely distributed in almost the entire Indo-Pacific, from East Africa to Tonga, north to southern Japan, and south to northern Australia, differs from Atherinomorus pinguis in having a wider midlateral band (the lower margin reaching to almost the center of the fourth scale row at level of the anal fin origin vs. the lower margin reaching to the ventral end of the third scale row in Atherinomorus pinguis) and more numerous midlateral scales (40–44 vs. 38–41 in Atherinomorus pinguis). Atherina morrisi Jordan and Starks, 1906, Hepsetia pinguis mineri Nichols and Roemhild, 1951, Pranesus capricornensis Woodland, 1961, Pranesus maculatus Taylor, 1964, and Pranesus pinguis ruppelli Smith, 1965, are regarded as junior synonyms of Atherinomorus lacunosus. Atherinomorus pinguis is also widely distributed in the Indo-West Pacific, from East Africa to northern Australia and north to southern Japan. Atherina pectoralis Valenciennes, 1835, is considered a junior synonym of Atherinomorus pinguis. Supplementary material to this paper is available in electronic format at     

<Emphasis Type="Italic">FORMOSA</Emphasis> controls cell division and expansion during floral development in <Emphasis Type="Italic">Antirrhinum</Emphasis><Emphasis Type="Italic">majus</Emphasis>

Control of organ size is the product of coordinated cell division and expansion. In plants where one of these pathways is perturbed, organ size is often unaffected as compensation mechanisms are brought into play. The number of founder cells in organ primordia, dividing cells, and the period of cell proliferation determine cell number in lateral organs. We have identified the Antirrhinum FORMOSA (FO) gene as a specific regulator of floral size. Analysis of cell size and number in the fo mutant, which has increased flower size, indicates that FO is an organ-specific inhibitor of cell division and activator of cell expansion. Increased cell number in fo floral organs correlated with upregulation of genes involved in the cell cycle. In Arabidopsis the AINTEGUMENTA (ANT) gene promotes cell division. In the fo mutant increased cell number also correlates with upregulation of an Antirrhinum ANT-like gene (Am-ANT) in inflorescences that is very closely related to ANT and shares a similar expression pattern, suggesting that they may be functional equivalents. Increased cell proliferation is thought to be compensated for by reduced cell expansion to maintain organ size. In Arabidopsis petal cell expansion is inhibited by the BIGPETAL (BPE) gene, and in the fo mutant reduced cell size corresponded to upregulation of an Antirrhinum BPE-like gene (Am-BPE). Our data suggest that FO inhibits cell proliferation by negatively regulating Am-ANT, and acts upstream of Am-BPE to coordinate floral organ size. This demonstrates that organ size is modulated by the organ-specific control of both general and local gene networks. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Phylogenetic placement of <Emphasis Type="Italic">Marasmiellus juniperinus</Emphasis>

Recent collections and the type specimen of Marasmiellus juniperinus, the type species of the genus, were examined. Phylogenetic placement, based on ribosomal large subunit (LSU) and internally transcribed spacer (ITS) sequences, is within the lentinuloid clade, nested among Gymnopus taxa. This placement dictates genus name usage and phylogenetic position of other putative species of Marasmiellus. The mating system is tetrapolar.     

Three new species of <Emphasis Type="Italic">Luetzelburgia</Emphasis> (<Emphasis Type="Italic">Leguminosae</Emphasis>, <Emphasis Type="Italic">Papilionoideae</Emphasis>) from the caatinga of Bahia,Brazil and an identification key to all species of the genus

Three new species of the Brazilian genus Luetzelburgia Harms (Leguminosae, Papilionoideae, Sophoreae sensu lato (Vataireoid clade)) are described and illustrated under the names Luetzelburgia harleyi D. Cardoso, L. P. Queiroz &; H. C. Lima, L. neurocarpa D. Cardoso, L. P. Queiroz &; H. C. Lima and L. purpurea D. Cardoso, L. P. Queiroz &; H. C. Lima. All three occur in the caatinga vegetation of the State of Bahia, Eastern Brazil. A discussion of the morphological characters distinguishing the new species from their close relatives, as well as an identification key to all known species of Luetzelburgia, are presented.     

Novel<Emphasis Type="Italic"> eceriferum</Emphasis> mutants in<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis>

We conducted a novel non-visual screen for cuticular wax mutants in Arabidopsis thaliana (L.) Heynh. Using gas chromatography we screened over 1,200 ethyl methane sulfonate (EMS)-mutagenized lines for alterations in the major A. thaliana wild-type stem cuticular chemicals. Five lines showed distinct differences from the wild type and were further analyzed by gas chromatography and scanning electron microscopy. The five mutants were mapped to specific chromosome locations and tested for allelism with other wax mutant loci mapping to the same region. Toward this end, the mapping of the cuticular wax (cer) mutants cer10 to cer20 was conducted to allow more efficient allelism tests with newly identified lines. From these five lines, we have identified three mutants defining novel genes that have been designated CER22, CER23, and CER24. Detailed stem and leaf chemistry has allowed us to place these novel mutants in specific steps of the cuticular wax biosynthetic pathway and to make hypotheses about the function of their gene products.Abbreviations EMS Ethyl methane sulfonate - SEM Scanning electron microscopy - SSLP Simple sequence length polymorphism - WT Wild type     

The ontogenetic basis for floral diversity in <Emphasis Type="Italic">Agonis</Emphasis>, <Emphasis Type="Italic">Leptospermum</Emphasis> and <Emphasis Type="Italic">Kunzea</Emphasis> (Myrtaceae)

Floral development in three species each of Leptospermum and Kunzea, and one species of Agonis, is described and compared. Differences in the number of stamens and their arrangement in the flower at anthesis are determined by the growth dynamics of the bud. In Leptospermum, early expansion of the bud is predominantly in the axial direction and causes the stamen primordia to be initiated in antepetalous chevrons. In Kunzea, early expansion occurs predominantly in the lateral direction and successive iterations of stamen primordia are inserted alternately at more or less the same level. In both genera, further expansion in the lateral plane spreads the stamens into a ring around the hypanthium. Agonis flexuosa is similar to Leptospermum. Other variable factors include the timing at which stamen initiation commences (earlier in Leptospermum than Kunzea), the duration of stamen initiation (hence the total number of stamens produced – varies within genera), and very late differential expansion that forces stamens into secondary antesepalous groups in A. flexuosa and L. myrsinoides.The authors thank Dr H. Toelken for kindly providing some material and the impetus for this project. This research was supported by Australian Research Council grant AS19131815.     

<Emphasis Type="Italic">tantalus</Emphasis>, a potential link between <Emphasis Type="Italic">Notch</Emphasis> signalling and chromatin-remodelling complexes

The tantalus (tan) gene encodes a protein that interacts specifically with the Polycomb/trithorax group protein Additional sex combs (ASX). Both loss-of-function and gain-of-function mutations in tan cause tissue-specific defects in the eyes, wing veins and bristles of adult flies. As these defects are also typical for components of the Notch (N) signalling pathway, we wished to determine if TAN interacts with this pathway. Through careful examination of ectopic tan phenotypes, we find that TAN specifically disrupts all three major processes associated with the N signalling pathway (boundary formation, lateral inhibition, and lineage decisions). Furthermore, ectopic tan expression abolishes expression of two N target genes, wingless (wg) and cut, at the dorsal-ventral boundary of the wing. An interaction between tan and N was also observed using a genetic assay that previously detected interactions between tan and Asx. The previously observed ability of TAN to move between the cytoplasm and nucleus, and to associate with DNA, provides a potential mechanism for TAN to respond to N signalling.Edited by P. Simpson     

Phylogeny of Chinese <Emphasis Type="Italic">Polystichum</Emphasis> (Dryopteridaceae) based on chloroplast DNA sequence data (<Emphasis Type="Italic">trnL-F</Emphasis> and <Emphasis Type="Italic">rps4-trnS</Emphasis>)

Polystichum is one of the largest and most taxonomically complex fern genera in China. The evolutionary relationships of Chinese Polystichum and related genera, and the relationship between our Polystichum phylogeny and ecogeographic distribution, were tested by the use of DNA sequence data. Fifty-one species of Polystichum and 21 species in allied genera were sequenced for the plastid intergenic spacers rps4-trnS and trnL-F. Maximum parsimony and Bayesian phylogenetic analyses of both individual and combined data sets showed that Chinese Polystichum as commonly recognized was paraphyletic: one clade (the CCPC clade) included Cyrtomidictyum lepidocaulon, two Cyrtogonellum species, three Cyrtomium species, and a small number of Polystichum species usually occurring on limestone. A second clade, Polystichum sensu stricto, included the remainder of the Polystichum species; these often occur on non-limestone substrates. The remaining Cyrtomium species formed the third clade. Three subclades resolved within Polystichum sensu stricto (s.s.) clade do not correspond with recent sectional classifications, and we outline the issues relevant to a new classification for the genus. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Molecular analysis of the<Emphasis Type="Italic"> CRINKLY4</Emphasis> gene family in<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis>

The maize (Zea mays L.) CRINKLY4 (CR4) gene encodes a serine/threonine receptor-like kinase that controls an array of developmental processes in the plant and endosperm. The Arabidopsis thaliana (L.) Heynh. genome encodes an ortholog of CR4, ACR4, and four CRINKLY4-RELATED (CRR) proteins: AtCRR1, AtCRR2, AtCRR3 and AtCRK1. The available genome sequence of rice (Oryza sativa L.) encodes a CR4 ortholog, OsCR4, and four CRR proteins: OsCRR1, OsCRR2, OsCRR3 and OsCRR4, not necessarily orthologous to the Arabidopsis CRRs. A phylogenetic study showed that AtCRR1 and AtCRR2 form a clade closest to the CR4 group while all the other CRRs form a separate cluster. The five Arabidopsis genes are differentially expressed in various tissues. A construct formed by fusion of the ACR4 promoter and the GUS reporter, ACR4::GUS, is expressed primarily in developing tissues of the shoot. The ACR4 cytoplasmic domain functions in vitro as a serine/threonine kinase, while the AtCRR1 and AtCRR2 kinases are not active. The ability of ACR4 to phosphorylate AtCRR2 suggests that they might function in the same signal transduction pathway. T-DNA insertions were obtained in ACR4, AtCRR1, AtCRR2, AtCRR3 and AtCRK1. Mutations in acr4 show a phenotype restricted to the integuments and seed coat, suggesting that Arabidopsis might contain a redundant function that is lacking in maize. The lack of obvious mutant phenotypes in the crr mutants indicates they are not required for the hypothetical redundant function.     

Sequence evolution and sex-specific expression patterns of the C class floral identity gene, <Emphasis Type="Italic">SpAGAMOUS</Emphasis>, in dioecious <Emphasis Type="Italic">Spinacia oleracea</Emphasis> L

Development in dioecious cultivated spinach, Spinacia oleracea, is distinguished by the absence of alternative reproductive organ primordia in male and female flowers. Given the highly derived floral developmental program in spinach, we wished to characterize a spinach C class floral identity gene and to determine the patterns of sequence evolution as well as compare the spatial and temporal expression patterns with those of AGAMOUS. The isolated cDNA sequence clusters phylogenetically within the AGAMOUS/FARINELLI C class clade. In comparison with the SLM1 sequence from the related Silene latifolia, amino acid replacements are highly conservative and non-randomly distributed, being predominantly found in hinge regions or on exposed surfaces of helices. The spinach gene (SpAGAMOUS) appears to be exclusively expressed in reproductive tissues and not in vegetative organs. Initial expression of SpAGAMOUS is similar in male and female floral primordia. However, upon initiation of the first whorl organs, SpAGAMOUS becomes restricted to meristemic regions from which the reproductive primordia will develop. This results in an early gender-specific pattern. Thus, the spinach C class gene is differentially expressed prior to reproductive organ development and is, at least, correlated with, if not directly involved in, the sexual dimorphism in spinach.Electronic Supplementary Material Supplementary material is available for this article at     

An ultrastructural comparative study of the sperm of<Emphasis Type="Italic"> Hyla pseudopseudis</Emphasis>,<Emphasis Type="Italic"> Scinax rostratus</Emphasis>, and<Emphasis Type="Italic"> S. squalirostris</Emphasis> (Amphibia: Anura: Hylidae)

We make detailed comparisons of the ultrastructure of the spermatozoon among three species of the family Hylidae, Hyla pseudopseudis, Scinax rostratus, and S. squalirostris. The acrosome complex consists of two conical structures covering the nuclear rostrum, the acrosome vesicle, and the subacrosomal cone. The nucleus has a moderately condensed chromatin with a conical shape in longitudinal sections and a circular shape in cross-sections. In H. pseudopseudis, mitochondria are numerous and circular, and in S. rostratus and S. squalirostris there are fewer mitochondria that are more elongate in longitudinal and transverse sections. In H. pseudopseudis, the mitochondrial collar starts adjacent to the distal centriole, occupying the whole midpiece, whereas in both Scinax species the mitochondrial collar starts only at the posterior one-third of the midpiece. In both Scinax species, the presence of juxta-axonemal fiber, axial sheath, and axial fiber in the tail are seemingly plesiomorphic characters, widespread among bufonoid frogs. In H. pseudopseudis, however, the absence of axial fiber and axial sheath seems to be derived from the typical bufonoid condition. The differences between Hyla and Scinax sperm endorse the separation of the two genera and suggest that sperm ultrastructure can be a useful tool to investigate relationships at the intrafamily level.     

Ontogeny, anatomy and systematic significance of ovular structures in the `eucalypt group' (Eucalypteae, Myrtaceae)

Development of ovular structures in the `eucalypt group' (Eucalyteae, Myrtaceae) was investigated using scanning electron microscopy and light microscopy. Different modes of placental expansion account for the variation in initiation and arrangement of ovules and ovulodes. In Eucalyptus sensu stricto placental elongation proceeds proximally and the distally-positioned ovulodes develop from the first primordia initiated (congenital ovulodes). In Angophora, Corymbia, Arillastrum, Allosyncarpia and Stockwellia placental elongation is bi-directional and the ovulodes develop from last-initiated primordia (residual ovulodes). Bi-directional elongation and residual ovulodes are present in the outgroup taxon Lophostemon confertus and is presumably plesiomorphic. This suite of characters is potentially informative for resolving relationships for several key taxa: E. microcorys (subgenus Symphyomyrtus) has proximal placental expansion and congenital ovulodes, which is consistent with most Eucalyptus; E. curtisii (subgenus Gaubaea) has the plesiomophic condition and may be sister to all other species of Eucalyptus.