Micropropagation of four Gentiana species (G. lutea, G. cruciata, G. purpurea and G. acaulis)

The growth of axillary shoots was initiated on nodal stem segments, excised from aseptically grown seedlings of Gentiana acaulis L., G. cruciata L., G. lutea L. and G. purpurea L. In later subcultures, a basal callus tissue developed on the shoots, giving rise to de novo formed buds. Optimum benzyladenine and indoleacetic acid combinations for shoot development were established. They were slightly different in the four species. From 35-70% of shoots rooted spontaneously, except in G. lutea, in which adventitious roots were induced by applying naphthaleneacetic acid. It was conduded that the four Gentiana species were amenable to propagation in vitro. This revised version was published online in June 2006 with corrections to the Cover Date.     

On Chytridium braun,Diplochytridium N. G., and Canteria N. G. (Chytridiales)

Summary The operculate genus Chytridium Braun is emended and restricted to approximately 25 known species and several incompletely known members which lack an apophysis and develop much like species of Rhizophydium, insofar as the zoosporangium and absorbing or rhizoidal system are concerned, and form endobiotic or intramatrical resting spores. This interpretation corresponds fairly closely to Braun's diagnosis of the type species.A new genus, Diplochytridium, is established to segregate the former Chytridium species with an endobiotic or intramatrical apophysis or prosporangium and resting spores, which may develop sexually or asexually. Some of these species have a well-defined endo-exogenous alternation of growth and development in which the apophysis appears to function as a prosporangium. As interpreted here, Diplochytridium includes approximately 20 of the known species.Another new genus, Canteria, is created for a parasite of Mougeotia which Canter first described as a species of Phlyctidium but later found to develop endobiotic resting spores or zygospores by the fusion of conjugation tubes.     

Morphometrics of Globodera tabacum tabacum,G. t. virginiae,and G. t. solanacearum (Nemata: Heteroderinae)

A morphometric evaluation of second-stage juveniles (J2), males, females, cysts, and eggs of several isolates of the tobacco cyst nematode (TCN) complex, Globodera tabacum tabacum (GTT), G. t. virginiae (GTV), and G. t. solanacearum (GTS) is presented. Morphometrics of eggs, J2, and males are considerably less variable than of females and cysts. No measurements of eggs and J2 are useful for identification of the three subspecies. Distance from the median bulb and excretory pore to the head end in J2 and males is quite stable. Stylet knob width of males is useful for identifying GTV isolates and tail length in separating males of GTT isolates from GTV and GTS. Body length/width (L/W) ratio of females and cysts discriminates GTT from GTV and GTS; stylet knob width is an auxiliary character for identifying GTV. This subspecies complex has a continuum of values for the other characters. Data suggest a close relationship between GTV and GTS, which also occur in close proximity in Virginia.     

Pollination in Geonoma macrostachys and Three Congeners,G. acaulis,G. gracilis,and G. interrupta

A continuous 15 month study of the floral ecology of four syntopic understorey palm species of Genoma was conducted in Amazonian Peru lowland rainforest. The spicate inflorescences of G. macrostachys, G. acaulis and G. gracilis are strictly protandrous and the plants are functionally dioecious. Data suggest that in G. macrostachys and G. acaulis pollination is based on a mimicry system, the pistillate flowers mimicking the staminate ones in colour, shape and scent. Pollen-collecting meliponine bees (Hymenoptera, Apidae, Meliponinae) and pollen-feeding syrphid flies (Diptera, Syrphidae) which visit inflorescences during both sexual stages are the pollinators of G. macrostachys. Geonoma acaulis is pollinated by small pollen-feeding weevils (Coleoptera, Curculionidae, Derelomini) that visit male and female spikes. Additionally, in G. macrostachys another pollinator type, viz. euglossine bees (Hymenoptera, Apidae, Euglossinae), which are attracted and rewarded by both types of flowers may account for long-distance pollination. The palm G. gracilis shows a very distinct pollination system. Although opportunistic insect visitors are attracted to the inflorescences of this species it seems to be mainly anemophilous because pollen becomes powdery during an thesis. The branched inflorescences of G. interrupta are also protandrous, but unlike the other species of Geonoma observed, staminate and pistillate anthesis of individual flowers are, for the most, overlapping. A broad spectrum of visitors is attracted (bees, wasps, flies, and beetles), which all may act as pollinators. Outcrossing is especially encouraged during the purely female phase at the end of the flowering cycle when there are no more staminate flowers in the inflorescence. Effects on the reproductive biology and population structure of different pollination systems and breeding system are discussed.     

The comparative leaf anatomy of Goniothamus andersonii, G. macrophyllus, G. malayanus and G. velutinus

The leaf anatomy of Goniothalamus andersonii, G. macrophyllus, G. malayanus and G. velutinus from the peat swamps of Sarawak is compared. Sufficient anatomical differences exist to differentiate the four species. G. velutinus has many points of difference from the other three species and G. macrophyllus is readily identified by the presence of fibre-sclereids in the lamina mesophyll. G. andersonii and G. malayanus are similar to each other, but G. andersonii can be distinguished, in particular by the more pronounced multiple projections of the outer periclinal walls of the epidermal cells and the presence of thick and cutinised outer ends of the anticlinal walls of the epidermal cells.     

The comparative stem and root anatomy of Goniothalamus andersonii, G. macrophyllus, G. malayanus and G. velutinus (Annonaceae) from the peat swamps of Sarawak

The stem and root anatomy of G. andersonii, G. macrophyllus and G. velutinus from the peat-swamps of Sarawak is compared. Sufficient anatomical differences exist to differentiate the four species. The main distinguishing anatomical features between the stems are the cork cells, the nature and relative quantity of cortical sclereids and the presence or absence of lignified pith parenchyma. In differentiating between the roots of G. andersonii, G. macrophyllus and G. malayanus. the principal features are the cork cells and the relative quantity of cortical sclereids. In addition, the medullary rays of G. macrophyllus root in transverse sections are noticeably broader in the xylem towards the phloem than those of the other species.     

MORPHOLOGY AND MOLECULAR ANALYSES OF THREE TOXIC SPECIES OF GAMBIERDISCUS (DINOPHYCEAE): G. PACIFICUS, SP. NOV., G. AUSTRALES, SP. NOV., AND G. POLYNESIENSIS, SP. NOV.

Three new dinoflagellate species, Gambierdiscus polynesiensis, sp. nov., Gambierdiscus australes, sp. nov., and Gambierdiscus pacificus, sp. nov., are described from scanning electron micrographs. The morphology of the three new Gambierdiscus species is compared with the type species Gambierdiscus toxicus Adachi et Fukuyo 1979, and two other species: Gambierdiscus belizeanus Faust 1995 and Gambierdiscus yasumotoi Holmes 1998. The plate formula is: Po, 3′, 7", 6C, 8S, 5‴, 1p, 2". Culture extracts of these three new species displayed both ciguatoxin- and maitotoxin-like toxicities. The following morphological characteristics differentiated each species. 1) Cells of G. polynesiensis are 68–85 μm long and 64–75 μm wide, and the cell’s surface is smooth. They are identified by a large triangular apical pore plate (Po), a narrow fish-hook opening surrounded by 38 round pores, and a large, broad posterior intercalary plate (1p) wedged between narrow postcingular plates 2‴ and 4‴. Plate 1p occupies 60% of the width of the hypotheca. 2) Cells of G. australes also have a smooth surface and are 76–93 μm long and 65–85 μm wide in dorsoventral depth. They are identified by the broad ellipsoid apical pore plate (Po) surrounded by 31 round pores and a long and narrow 1p plate wedged between postcingular plates 2‴ and 4‴. Plate 1p occupies 30% of the width of the hypotheca. 3) Cells of G. pacificus are 67–77 μm long and 60–76 μm wide in dorsoventral depth, and its surface is smooth. They are identified by the four-sided apical pore plate (Po) surrounded by 30 round pores. A short narrow 1p plate is wedged between the wide postcingular plates 2‴ and 4‴. Plate 1p occupies 20% of the width of the hypotheca. These three newly described species were also characterized by isozyme electrophoresis and DNA sequencing of the D8–D10 region of their large subunit (LSU) rRNA genes. The consistency between species designations based on SEM microscopy and classification inferred from biochemical and genetic heterogeneities was examined among seven isolates of Gambierdiscus. Their classification into four morphospecies was not consistent with groupings inferred from isozyme patterns. Three molecular types could be distinguished based on the comparison of their LSU rDNA sequences. Although G. toxicus TUR was found to be more closely related to G. pacificus, sp. nov. than to other G. toxicus strains, the molecular classification was able to discriminate G. polynesiensis, sp. nov. and G. australes, sp. nov. from G. toxicus. These results suggest the usefulness of the D8–D10 portion of the Gambierdiscus LSU rDNA as a valuable taxonomic marker.     

Sperm cryopreservation in three species of endangered gazelles (Gazella cuvieri,G. dama mhorr,and G. dorcas neglecta)

Long-term storage of semen by cryopreservation, with high recovery rates on thawing, is essential for the establishment of genetic resource banks of endangered species. The purpose of the present study was to evaluate various diluents for the cryopreservation of spermatozoa from three species of gazelles (genus Gazella) in a captive breeding program. The diluents compared were Tes (N-tris(hydroxymethyl)methyl-2 aminoethane sulfonic acid)-Tris with 5% egg yolk and 6% glycerol (TEST) and Triladyl, yolk-citrate, Tris-trehalose, and Tris-lactose-all of them with 20% egg yolk and 6% (Triladyl) or 8% glycerol. Semen was obtained by electroejaculation from 12 G. cuvieri, 12 G. dama, and 13 G. dorcas males. Samples with less than 50% motile sperm, positive endosmosis, or acrosome integrity were not used. Diluted samples were loaded into 0.25-ml straws, cooled slowly to 5 degrees C over 1.5 h (-0.16 degrees C/min), equilibrated at that temperature for 2 h, frozen in nitrogen vapors for 10 min, and plunged into liquid nitrogen. Subsamples were assessed fresh, after refrigeration-equilibration, after freezing and thawing, and 2 h after thawing. Differences were seen between diluents, with best overall recovery rates after freezing and thawing found with Triladyl, TEST, and Tris-trehalose in G. cuvieri, TEST in G. dama, and Triladyl and TEST in G. dorcas. Differences were observed between species in the ability to withstand freezing and thawing, with best results seen in G. dorcas, intermediate results in G. dama, and worst results in G. cuvieri. These differences were inversely related to the average values of inbreeding of these populations. The underlying mechanism responsible for these differences may be a differential resistance to osmotic shock.     

Ribosomal DNA sequences of Glugea anomala, G. stephani, G. americanus and Spraguea lophii (Microsporidia): phylogenetic reconstruction

The microsporidian species Glugea anomala, G. stephani, G. americanus and Spraguea lophii were compared by using sequence data derived from their small subunit rDNA genes which were amplified by polymerase chain reaction and directly sequenced. These sequence data and published data of G. atherinae were analyzed and were used to infer a phylogenetic tree. The 5 microsporidian fish parasites appeared to be closely related. The higher sequence similarities demonstrated among G. anomala, G. stephani and G. atherinae suggest that these 3 parasites are in fact only 1 species of Glugea. Moreover, the higher sequence similarities between S. lophii and G. americanus support the transfer of the latter Glugea species into the genus Spraguea.     

Chromosome-banding study in Gerbillinae (Rodentia). I. Comparative analysis of Gerbillus poecilops, G. henleyi and G. nanus

An analysis was made of the R- and C-banding of chromosomes of the following Gerbillus species (Rodentia, Gerbillidae): G. poecilops (Taif, Saudi Arabia), G. henleyi (Oursi, Burkina Faso; Kairouan, Tunisia), and G. nanus (Taif and Hofuf, Saudi Arabia; Sind desert, Pakistan). Their karyotypes show the same chromosome number (2n = 52), but a different N.F.a. (number of autosomal arms), ranging from 58 to 62 in the species overall, and from 58 to 60 among G. nanus forms, due to pericentric inversions involving chromosomes 1 and 4. Moreover, large inter-individual variations of heterochromatin segments of chromosomes 25, X, and Y were found. Considering morphology and karyotype, G. poecilops and G. henleyi appear to be rather monomorphous species, whereas G. nanus produces a series of forms as a consequence of speciation, in which both chromosomal and morphological differentiations occur that do not correlate.