The sporophyte-gametophyte junction in the moss Acaulon muticum (Pottiaceae): EARLY STAGES OF DEVELOPMENT

The sporophyte-gametophyte junction in Acaulon muticum is composed of the sporophyte foot, the surrounding gametophyte vaginula, and an intervening placental space. At an early stage of development the foot has a large basal cell, characterized by extensive wall ingrowths beginning at the lowermost tip of the basal cell and extending along its tangential walls. Sporophyte cells in contact with the basal cell develop ingrowths on their outer tangential walls and on radial walls in contact with the basal cell. All sporophyte cells at this stage are characterized by numerous mitochondria, strands of endoplasmic reticulum, and dictyosomes, particularly in the cytoplasm adjacent to areas of extensive wall development. Plastids typically contain abundant starch reserves. As development proceeds, wall ingrowths become more extensive on all walls in the sporophyte foot but are never found on the upper wall of the basal cell in contact with the remainder of the sporophyte. Plastids in the foot contain fewer starch reserves later in development. Wall ingrowths are not visible in the cells of the gametophyte vaginula until well after extensive development has occurred in the sporophyte foot. Stacks or layers of endoplasmic reticulum are characteristic of the cells of the gametophyte vaginula, along with numerous mitochondria, dictyosomes, and well-developed plastids. Starch reserves typically are less abundant in cells of the gametophyte. The early development of extensive wall elaborations in the cells of the sporophyte foot, and particularly in the basal cell, may favor the rapid movement of water and nutrients from the gametophyte into the sporophyte at a time when rapid development in this minute, ephemeral moss is critical.     

Spermiogenesis in the cestode Corallobothrium solidum Fritsch, 1886 (Proteocephalidea: Corallobothriinae)

Spermiogenesis of Corallobothrium solidum Fritsch 1886, has been investigated by transmission electron microscopy. The zone of differentiation contains the two centrioles, each with one thin root, being orientated in the same plane only when a single intercentriolar body (ICB) appears between them. A median cytoplasmic process (MCP) develops distally to the two flagella, which are of unequal length, get longer and rotate towards the MCP. The nucleus penetrates into the spermatid body after the fusion of both flagella with the MCP has started. Flagellar roots occur occasionally in some spermatids. New for the Eucestoda are the following findings: 1. cortical microtubules (CMs) are arranged in two short parallel rows in one‐axoneme region of some spermatids; 2. the crested body of spermatid consists either of electron‐dense tubular elements and interposes itself between CMs, or it is rather homogeneous and situated more peripherally above one continuous semicircle of CMs. The present results support previous data that the type of spermiogenesis in proteocephalideans resembles mostly that observed in tetraphyllideans (Onchobothriidae and Phyllobothriidae), thus supporting the view of a close phylogenetic relationship of tetraphyllidean and proteocephalidean cestodes.     

alpha-D-Glucuronosyl-(1-->3)-L-galactose,an unusual disaccharide from polysaccharides of the hornwort Anthoceros caucasicus

Acid hydrolysis of cell wall-rich material from thalli of the hornwort Anthoceros caucasicus yielded substantial amounts of an unusual disaccharide (1). Hydrolysis of 1 yielded only GlcA, Gal and unhydrolysed 1. Compound 1 was identified as alpha-D-GlcpA-(1-->3)-L-Gal by 1H and 13C NMR spectroscopic analysis and by the susceptibility of its monosaccharide units to phosphorylation by enantiomer-specific kinases. Compound 1 was not detected in acid hydrolysates of other land plants including mosses, leafy and thalloid liverworts, lycopodiophytes and euphyllophytes; it was also absent from charophytes. The Anthoceros polysaccharide that yields 1 was partially extractable in cold aqueous buffer (pH 4.7) and Na(2)CO(3), but not in EDTA or NaOH, suggesting that it was not a typical pectin or hemicellulose. The yield of 1 from various polysaccharide fractions correlated with the yields of Xyl, suggesting a previously unreported polymer containing D-GlcA, L-Gal and Xyl. The existence of a unique polysaccharide in an evolutionarily isolated plant (Anthoceros) supports the view that major steps in plant phylogeny were accompanied by significant changes in cell wall composition.     

Chloroplast, mitochondrial, and nuclear microsatellites from the southern Appalachian hornwort, Nothoceros aenigmaticus (Dendrocerotaceae)

? Premise of the study: New microsatellite primers were developed for testing genetic differentiation within Nothoceros aenigmaticus and their potential use in other Nothoceros species. The microsatellites are designed to investigate partitioning of genetic variation in a taxon with a peculiar sex allopatry in the southern Appalachian Mountains and relationships with conspecific sexual populations from Mexico. ? Methods and Results: We used two methods for microsatellite development: an enriched library and second-generation shotgun sequence reads. From these two methods, a total of nine primer pairs were selected and tested on 89 southern Appalachian N. aenigmaticus accessions, nine Mexican accessions, and 16 N. vincentianus accessions. Three mitochondrial loci were recovered from the enriched library method and six loci from 454 shotgun sequencing: three were from the chloroplast and three from the nucleus. The primers amplified repeats with two to 20 alleles per locus. ? Conclusions: New microsatellite primers were developed for testing genetic differentiation within N. aenigmaticus and potentially for use in other Nothoceros species. We present one of the first reports of highly polymorphic mitochondrial microsatellites in plants.     

Massive ribonucleoprotein bodies in gametophyte vegetative cells of the mossTimmiella barbuloides (Brid.) Moenk

Summary Vegetative cells of the gametophyte phase of the mossTimmiella barbuloides (Pottiales) are characterized by large cytoplasmic bodies of spherical shape (SBs) whose ribonucleoprotein composition is cytochemically demonstrated. SBs seem to be derived from massive aggregation of cytoplasmic ribosomes, with possible participation by rough endoplasmic reticulum elements. SBs have been found in stereids, parenchymatous cells and young hydroids of the gametophyte stem, and in euricysts of the leaf nerve. The SBs develop early in the course of cell differentiation and, once formed, persist until advanced stages of cell senescence.     

Protein tyrosine phosphatase interacting protein 51 (PTPIP51) mRNA expression and localization and its in vitro interacting partner protein tyrosine phosphatase 1B (PTP1B) in human placenta of the first, second, and third trimester

The cellular localization of protein tyrosine phosphatase 51 (PTPIP51) and its in vitro interacting partner protein tyrosine phosphatase 1B (PTP1B) was studied in human placentae of different gestational stages. The expression of PTPIP51 protein and mRNA was observed in the syncytiotrophoblast and cytotrophoblast layer of placentae from the first, second, and third trimesters. In contrast, PTP1B expression was restricted to the syncytiotrophoblast during all gestational stages. Cells of the cytotrophoblasts and parts of the syncytiotrophoblasts expressing high amounts of PTPIP51 were found to execute apoptosis as shown by TdT-mediated dUTP-biotin nick end labeling assay, cytokeratin 18f, and caspase 3 expression. PTPIP51 could also be traced in the endothelium and smooth muscle cells of placental arterial and venous vessels, identified by double immunostainings with antibodies directed against van Willebrand factor and alpha-smooth muscle actin. Some of these cells showing a high PTPIP51 reactivity were Ki67 positive, indicating proliferation. Additionally, a small population of placental CD14-positive macrophages and mesenchymal cells within the villous stroma were detected as PTPIP51 positive. Our data suggest that both proteins, PTPIP51 and PTP1B, play a role in differentiation and apoptosis of the cytotrophoblast and syncytiotrophoblast, respectively. Moreover, PTPIP51 may also serve as a cellular signaling partner in angiogenesis and vascular remodeling.     

Ultrastructure of the trophosome, a food-storage organ in Gastromermis boophthorae (Nematoda: Mermithidae)

The ultrastructure of the trophosome of Gastromermis boophthorae (Welch & Rubzov, 1965) is described at selected points in the life cycle of this mermithid nematode. The trophosome is grossly modified for the assimilation and storage of nutrients, and comparison is made with a generalised nematode intestine, with which it may be homologous. At its maximum development the trophosome is a solid syncytial cylinder which almost fills the body cavity of the nematode. Internally it is packed with nutrient reserves, whilst the outer cytoplasmic layer exhibits features indicative of transport functions. The trophosome is linked to the hypodermis of the body wall by radially-orientated cytoplasmic bridges, and the possible significance of this arrangement is discussed in relation to the nutritional function of the trophosome.     

The cytology of Actinidia, Saurauia and Clematoclethra (Actinidiaceae)

Meiosis and mitosis of six Chinese Actinidiaceae were studied: Saurauia tristyla DC., S. miniata C. F. Liang & Y. S. Wang, Actinidia chinensis Plach., A. deliciosa (Cheval.) C. F. Liang & A. R. Ferguson, A. indochinensis Merr. and Clematoclethra lasioclada Maxim. The chromosome numbers of Saurauia tristyla and S. miniata were 2 n  = 6 x  = 78, establishing a base chromosome number of x  = 13 in the genus, differing from the previous report of x  = 15. The chromosome number of Clematoclethra was first reported to be 2 n  = 4 x  = 48 ( x  = 12), while that of Actinidia was x  = 29, consistent with previous reports. The base chromosome number of Clematoclethra ( x  = 12) was derived from an aneuploid decrease from Saurauia ( x  = 13). Actinidia (x  = 29) was derived from the palaeotetraploid ( x  = 14), which was formed through the increase of the basic chromosome number x  = 13 to x  = 14 by aneuploidy and through the breakage of a centromere to add one more new chromosome. The chromosome data in Actinidia were consistent with the geographical and morphological evidence for the evolution of the three genera. The tropical American and Asian disjunct distribution pattern and the diversity of base chromosome numbers of Saurauia further support the probability that the genus was an early divergent from a common ancestor of Actinidia and Clematoclethra .  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 369–374.     

白细胞介素—2—绿脓杆菌外毒素融合蛋白的分高纯化及其复性研究

表达的白细胞介素－２－绿脓杆菌外毒素（ＩＬ－２－ＰＥ）融合蛋白以包含体形式存在于宿主菌中,为分离纯化表达产物提供了方便,但因需进行复性,也增加了后处理的难度．我们采用４ｍｏｌ／Ｌ尿素、０．５％ＴｒｉｔｏｎＸ－１００的１×ＰＢＳ洗涤包含体两遍,再经ＳｅｐｈａｃｒｙｌＳ－３００分子筛及ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＦＦ阴离子交换柱层析后,获得的融合蛋白纯度可达９０％～９５％。此外,我们从ＧＳＳＧ浓度、Ｌ－精氨酸浓度、复性蛋白质的起始浓度、复性液的ｐＨ值、复性温度及复性时间等参数入手,系统地研究了融合蛋白的复性条件,探索到了ＩＬ－２－ＭＥ４０和ＩＬ－２－ＰＥ６６４Ｇｌｕ融合蛋白复性的最适条件。     

Genetic and morphological divergence between populations of the flatfish Platichthys flesus (L.) (Pleuronectidae)

Populations of the flounder, Platichthys flesus , were screened for electrophoretically detectable protein variation at up to 37 loci. Atlantic and North Sea populations (subspecies flesus ) were genetically very similar to one another ( I >0.99) but different from Adriatic (subspecies italicus ) and Black Sea (subspecies luscus ) populations. The values for genetic identity between subspecies were around 0.9. Diagnostic loci enabled specimens of flesus, italicus and luscus to be differentiated from one another. Samples of flesus showed two to three times the heterozygosity levels of italicus or luscus , consistent with the greater population size of flesus . Morphological comparisons enabled populations to be categorized to subspecies. Platichthys flesus italicus and luscus are valid subspecies: the terms are not synonyms. The genetic distance data provide an estimate of divergence time of the three subspecies at about 2 My ago, an estimate that seems reasonable in the light of present knowledge concerning the recent geological histories of the Mediterranean and Black Sea basins.     

Observations on the mature gametophyte of Phylloglossum (Lycopodiaceae)

Mature, gametangia-bearing photosynthetic gametophytes of Phylloglossum can be grown from the nonphotosynthetic, cylindrical, negatively gravitropic immature stages in illuminated axenic culture on a nutrient medium with or without 0.2% glucose. The gametangial-bearing region of these gametophytes, the photosynthetic crown, grows horizontally from the apex of the immature cylindrical stage. At maturity the photosynthetic crown is thickened and bilaterally symmetrical. It is usually narrow and bean-shaped with dorsal and ventral regions. Occasionally, the ventral region becomes thicker and the crown is deltoid in cross section. The dorsal edge or ridge of the crown is the gametangial region with archegonia and antheridia. The gametangia are often hidden because they are interspersed among numerous uni- or bicellular paraphyses. The lateral surfaces below the gametangial region lack outgrowths, and the ventral region is covered with long rhizoids. The apical meristem at the anterior end of the crown is overarched by the developing dorsal tissue of the gametangial ridge. The lower derivatives of the meristem form the ventral region with rhizoids. Phylloglossum gametophytes are unusual because their mode of organic nutrition changes from mycorrhizal to photosynthetic during ontogeny.     

Identification and localization of a protein immunologically related to Caltractin (Centrin) in the Myonemes and Membranelles of the Heterotrich ciliate Stentor coeruleus

The contractile properties of the myonemes of Stentor are very similar to caltractin (centrin)-containing fibers of other organisms. We investigated whether the calcium-binding protein caltractin was present in Stentor by using three different antibodies to caltractin or caltractin-related proteins, in conjunction with immunofluorescence microscopy and protein blotting. Immunofluorescence demonstrated that a protein immunologically similar to caltractin is present in the myonemes and in the bases of the membranelles of Stentor. The localization to the myonemes is observed in intact cells, osmotically lysed cells, and isolated cortices. Double-label immunofluorescence with anti-alpha-tubulin and anti-caltractin antibodies showed that the fluorescence in the myonemes was not in the overlying Km fibers. The myonemes in the posterior one-third of the cell appear as thick fibers with no cross-bridging. They become thinner as they approach the anterior end of the cell and show extensive cross-bridging here. Staining in the bases of the membranelles shows a distinct comma-like immunofluorescence pattern similar to that seen with protargol-stained cells and SEM views of the membranellar band reported by others. Western blots demonstrated that the caltractin-like protein in Stentor has an apparent molecular weight of 23 kDa compared with the 20-kDa protein from Chlamydomonas and is a calcium-binding protein.     

Variation in gametophyte dominance in populations of Chondrus verrucosus (Gigartinaceae,Rhodophyta)

We describe the abundance, including spatial and temporal variability, of phases of the isomorphic Chondrus verrucosus Mikami from Japan. Chondrus verrucosus occurred in a dense (～90% cover) and temporally stable bed on a small, isolated rocky outcrop (Oyakoiwa) in Shizuoka Prefecture. Small vegetative fronds were always much more abundant than large vegetative and fertile fronds over the spring to late summer periods in 1999 and 2000. Over the same period, fertile carposporophytic fronds were generally more abundant than fertile tetrasporophytic fronds, and fertile male fronds appeared infrequently at low densities. Using the resorcinol‐acetal test, we determined the proportion of gametophytes and tetrasporophytes in three populations of C. verrucosus: Oyakoiwa and Noroshi (Shizuoka) in the summers of 1999 and 2000 and Kamehana Point (Miyagi) in autumn 2000. All populations had a significantly higher proportion of gametophytes than tetrasporophytes in both years, although gametophytic proportions were lower at Noroshi (～70%) than at Oyakoiwa (～80%) and Kamehana Point (～97%). However, examination of all isolated individuals sampled on Noroshi showed equal proportions of each phase in 1999, but gametophyte dominance (74%) in 2000. Differences in dispersal and spore production between phases are discussed as mechanisms potentially contributing to variation in gametophyte dominance.     

Placental insulin‐like growth factor II (IGF‐II) and its relation to litter size in the common marmoset monkey (Callithrix jacchus)

The primate placenta produces a wide variety of hormones throughout gestation that regulate placental function and fetal growth. One such hormone is insulin‐like growth factor‐II (IGF‐II), a peptide implicated in cell division, differentiation, and amino acid transport. IGF‐II concentrations were measured in 23 common marmoset (Callithrix jacchus) term placentas from twin and triplet litters in order to determine whether previously described differences in fetoplacental phenotype such as placental and litter mass and placental surface area were related to differences in endocrine function. IGF‐II was extracted from frozen tissue samples and measured using an enzyme‐linked immunosorbent assay kit designed for human tissue, which was validated for marmoset placenta. IGF‐II concentrations were not related to placental or litter mass, and twin and triplet placentas did not differ in total concentration. However, per individual fetus, triplets were associated with a significant 42% reduction in IGF‐II concentration (P=0.03), and IGF‐II concentration per gram of fetal mass was a third lower in triplet litters. The triplet placenta exhibits a global expansion of the surface area which was contrasted by a per unit area reduction in IGF‐II concentration (r=?0.75, P=0.01), a pattern that explains why twin and triplet placentas overall did not differ in concentration. Per fetus, triplet pregnancies are associated with relatively less maternal mass, placental mass and microscopic surface area suggesting that the intrauterine growth of triplets is supported by systems that increase the efficiency of nutrient transfer. The finding that individual triplet fetuses are also associated with significantly lower IGF‐II concentrations is consistent with the view that the marmoset fetoplacental unit exhibits a flexible pattern of placental allocation and metabolism. Plasticity in placental endocrine and metabolic function is likely to play an important role in the ability of the fetus to sense and accommodate the intrauterine environment and, by extension, the external ecology. Am. J. Primatol. 71:969–975, 2009. © 2009 Wiley‐Liss, Inc.     

濒危植物红花木莲大孢子发生和雌配子体发育的研究

研究了濒危植物红花木莲（Manglietia insignisBl)的大孢子发生和雌配子体的发育,红花木莲每朵花心皮数为48-73枚,每心皮内胚珠数4-8颗不等,多数是6颗或8颗,胚朱倒生,双珠被,厚珠心,大孢子四分体线形排列,功能大孢子位于合点端,胚囊发育为蓼型。胚珠具珠孔塞,成熟胚囊的卵细胞败育率高达79.1%。这是造成栽培红花木莲结籽率低的主要原因,较低的结籽率影响了红花木莲迁地保护的有效性。     

A precursor of the reserve-protein, phaseolin, is transiently associated with the endoplasmic reticulum of developing Phaseolus vulgaris cotyledons

Developing cotyledons of Phaseolus vulgaris L. were labeled for 30 min with [³H] amino acids, homogenized, and the proteins fractionated on sodium dodecylsulfate (SDS) polyacrylamide gels. Fluorographs of these gels showed that the polypeptides of phaseolin, the major reserve protein of P. vulgaris, were synthesized as precursors which could be distinguished from the polypeptides of mature phaseolin by their slightly lower mobility. When extracts of cotyledons labeled for 45 min with [³H] amino acids were fractionated on isopynic sucrose gradients, radioactive phaseolin banded at the same density (1.14 g cm^-3) as the endoplasmic reticulum (ER)-marker enzyme NADH-cytochrome c reductase. Fractionation in the presence of 3 mM MgCl₂ indicated that the newly-synthesized phaseolin was associated with the rough ER. Pulse-chase experiments showed that phaseolin was transiently associated with the ER, and later accumulated in the protein bodies. Treatment of isolated ER with proteinase K showed that phaseolin polypeptides were degraded only if Triton X-100 was present, indicating that phaseolin was membrane-protected, probably enclosed within the vesicles. ER-associated phaseolin associated to an 18S form at pH 4.5 in the presence of 0.3 M NaCl and 100 mM sodium acetate. The polypeptides of ER-associated phaseolin had a slightly lower mobility on SDS-gels than polypeptides of protein body phaseolin. ER-associated phaseolin had a carbohydrate content of 6.8%, while protein body-derived phaseolin had a carbohydrate content of 6.2%. When cotyledons were labeled simultaneously with [¹⁴C] amino acids and [³H] glucosamine or with [¹⁴C] amino acids and [³H] mannose, the [³H]/[¹⁴C] ratio of ER-derived phaseolin was similar to that of protein body derived phaseolin, indicating that the faster mobility on SDS-gels was not due to the detachment of carbohydrate. Experiments in which the carbohydrate side chains were removed with endoglycosidase H, and the resulting polypeptides subjected to electrophoresis in SDS-gels showed that the differential mobility of the glycopolypeptides of phaseolin resided in their polypeptide chains.     

霍氏啮小蜂外生殖器及其感器的超微结构

霍氏啮小蜂Tetrastichus howardi(Olliff)是许多鳞翅目害虫的群居性蛹寄生蜂。为分析霍氏啮小蜂成虫产卵器和交配器上与成虫产卵和交配有关的感受器类型,本文利用扫描电镜对霍氏啮小蜂成虫产卵器和交配器的超微结构进行了观察。在霍氏啮小蜂成虫产卵器上共发现了7种感受器,分别是毛形感器1(TS1)、毛形感器2(TS2)、刺形感器2(CH2)、分泌毛孔(SP)、栓锥形感器1(SS1)、栉齿状感器(DS)、浅凹形感器(SD)和火山形感器(VS)。交配器上共发现6种感受器官,分别是栓锥形感器2(SS2)、火山形感器(VS)、表皮孔(CP)、鸡冠状结构(Crs)、钟形感器(CAS)和毛形感器3(TS3)。结果表明霍氏啮小蜂可通过产卵器和交配器上感受器获得与寄主和配偶相关的信息,进而调控产卵行为和交配行为。