Nest-mediated seed dispersal

Many plant seeds travel on the wind and through animal ingestion or adhesion; however, an overlooked dispersal mode may lurk within those dispersal modes. Viable seeds may remain attached or embedded within materials birds gather for nest building. Our objective was to determine if birds inadvertently transport seeds when they forage for plant materials to build, insulate, and line nests. We also hypothesized that nest-mediated dispersal might be particularly useful for plants that use mating systems with self-fertilized seeds embedded in their stems. We gathered bird nests in temperate forests and fields in eastern North America and germinated the plant material. We also employed experimental nest boxes and performed nest dissections to rule out airborne and fecal contamination. We found that birds collect plant stem material and mud for nest construction and inadvertently transport the seeds contained within. Experimental nest boxes indicated that bird nests were not passive recipients of seeds (e.g., carried on wind), but arrived in the materials used to construct nests. We germinated 144 plant species from the nests of 23 bird species. A large proportion of the nest germinants were graminoids containing self-fertilized seeds inside stems—suggesting that nest dispersal may be an adaptive benefit of closed mating systems. Avian nest building appears as a dispersal pathway for hundreds of plant species, including many non-native species, at distances of at least 100–200 m. We propose a new plant dispersal guild to describe this phenomenon, caliochory (calio = Greek for nest).     

24-Hour cooled storage of equine embryos

Equine embryos were collected by transcervical uterine flush 7 d after ovulation. The flush solution was Dulbecco's phosphate buffered saline (PBS) with 1% newborn calf serum and penicillin-streptomycin. Each embryo was washed in modified Dulbecco's PBS with 1% newborn calf serum and 0.4% bovine serum albumin, and placed in 4-ml polystyrene test tube containing this same medium. Embryos were packaged in a commercial semen transport container which cooled (-0.3 degrees C/min) and maintained the embryo at 4 to 6 degrees C. After 24 h, 16 embryos were transcervically transferred into recipient mares. Of the 16 embryos, six were detected as vesicles by ultrasonography at 14 d of pregnancy, of which three were carried to term and resulted in live, normal foals. Sixteen control embryos were directly transferred without prior storage and resulted in five foals.     

Genetic evidence for predominantly hydrochoric gene flow in the invasive riparian plant Impatiens glandulifera (Himalayan balsam)

Background and Aims

Riparian systems are prone to invasion by alien plant species. The spread of invasive riparian plants may be facilitated by hydrochory, the transport of seeds by water, but while ecological studies have highlighted the possible role of upstream source populations in the establishment and persistence of stands of invasive riparian plant species, population genetic studies have as yet not fully addressed the potential role of hydrochoric dispersal in such systems.

Methods

A population genetics approach based on a replicated bifurcate sampling design is used to test hypotheses consistent with patterns of unidirectional, linear gene flow expected under hydrochoric dispersal of the invasive riparian plant Impatiens glandulifera in two contrasting river systems.

Key results

A significant increase in levels of genetic diversity downstream was observed, consistent with the accumulation of propagules from upstream source populations, and strong evidence was found for organization of this diversity between different tributaries, reflecting the dendritic organization of the river systems studied.

Conclusions

These findings indicate that hydrochory, rather than anthropogenic dispersal, is primarily responsible for the spread of I. glandulifera in these river systems, and this is relevant to potential approaches to the control of invasive riparian plant species.     

A Structural Element within the HUWE1 HECT Domain Modulates Self-ubiquitination and Substrate Ubiquitination Activities

E3 ubiquitin ligases catalyze the final step of ubiquitin conjugation and regulate numerous cellular processes. The HECT class of E3 ubiquitin (Ub) ligases directly transfers Ub from bound E2 enzyme to a myriad of substrates. The catalytic domain of HECT Ub ligases has a bilobal architecture that separates the E2 binding region and catalytic site. An important question regarding HECT domain function is the control of ligase activity and specificity. Here we present a functional analysis of the HECT domain of the E3 ligase HUWE1 based on crystal structures and show that a single N-terminal helix significantly stabilizes the HECT domain. We observe that this element modulates HECT domain activity, as measured by self-ubiquitination induced in the absence of this helix, as distinct from its effects on Ub conjugation of substrate Mcl-1. Such subtle changes to the protein may be at the heart of the vast spectrum of substrate specificities displayed by HECT domain E3 ligases.     

Agreement between measures of total motility and membrane integrity in stallion sperm

Increasing seminal plasma concentrations in extended stallion semen were utilized to model decreasing sperm motility over time. Level of agreement was determined between flow cytometric measurement of sperm membrane integrity, using a combination of SYBR-14 and propidium iodide, and computer-assisted analysis of sperm motility. Values for total sperm motility (TMOT;%) and membrane integrity (SMI;%) were similar (∼80%) at Time 0 within all sperm treatments. However, TMOT was lower than SMI after 24 and 48 h of storage in treatments with >20% seminal plasma. At Time 0, agreement (bias and absolute difference) between TMOT and SMI was high (-0.7 and 5.6%, respectively), but decreased after 24 (10.8 and 15.1%, respectively) and 48 h (23.0 and 23.8%, respectively) of cooled storage as motility declined more rapidly than SMI. We concluded that TMOT and SMI measured separate aspects of sperm quality.     

Replication process of the parvovirus H-1. VIII. Partial denaturation mapping and localization of the replication origin of H-1 replicative-form DNA with electron microscopy.

Partial denaturation mapping, restriction endonuclease digestion, and electron microscopy were used to determine which end of the linear duplex replicative-form (RF) DNA molecule contains the origin of RF replication for the parvovirus H-1. This origin was localized within approximately 300 base pairs of the arbitrarily designated right end of the RF DNA, in the EcoRI or HaeII-A fragment. Based on denaturation behavior in formamide, the right end was also found to have a relatively high guanine plus cytosine content, whereas the region adjacent to the left terminus of the RF DNA molecule was adenine plus thymine rich.     

Expression of phosphatidylethanolamine-binding protein in the male reproductive tract: Immunolocalisation and expression in prepubertal and adult rat testes and epididymides

Phosphatidylethanolamine-binding protein (PBP) has been described previously in the male reproductive tract, where it has been implicated in the biogenesis and maintenance of antigen segregation of membranes. In the present study we have used a specific antiserum to PBP to determine its expression and localisation in the adult and prepubertal rat testis and epididymis by Western blotting and immunohistochemistry. In the adult rat testis, PBP was localised to step 17–19 elongating spermatids, residual bodies, and interstitial Leydig cells. In the adult epididymis, PBP was localised to epithelial cells of the caput, corpus, and cauda regions and to the cytoplasmic droplets of spermatozoa in the lumen of the initial segment, caput, and corpus epididymidis. In prepubertal animals, PBP was expressed in both testes and epididymides from day 1 and day 3 postpartum, respectively (day 3 being the earliest epididymal tissue taken). In prepubertal testes, PBP was localised to Leydig cells from day 1 postpartum and was not detected in any other cell type until the differentiation of elongate spermatids, when it was detected in step 17–19 elongating spermatids. These data suggest that PBP may be involved in the organisation of sperm membranes during spermiogenesis. The presence of PBP in Leydig cells, however, suggests diverse roles for this protein as a lipid carrier or binding protein. Mol. Reprod. Dev. 49:454–460, 1998. © 1998 Wiley-Liss, Inc.     

Ca2+ and Calmodulin Dynamics during Photopolarization in Fucus serratus Zygotes

The role of Ca2+ in zygote polarization in fucoid algae (Fucus, Ascophyllum, and Pelvetia species) zygote polarization is controversial. Using a local source of Fucus serratus, we established that zygotes form a polar axis relative to unilateral light (photopolarization) between 8 and 14 h after fertilization (AF), and become committed to this polarity at approximately 15 to 18 h AF. We investigated the role of Ca2+, calmodulin, and actin during photopolarization by simultaneously exposing F. serratus zygotes to polarizing light and various inhibitors. Neither removal of Ca2+ from the culture medium or high concentrations of EGTA and LaCl3 had any effect on photopolarization. Bepridil, 3,4,5-trimethoxybenzoic acid 8-(diethylamino) octyl ester, nifedipine, and verapamil, all of which block intracellular Ca2 release, reduced photopolarization from 75 to 30%. The calmodulin antagonists N-(6-aminohexyl)-5-chloro-L-naphthalenesulfonamide and trifluoperazine inhibited photopolarization in all zygotes, whereas N-(6-aminohexyl)-L-naphthalenesulfonamide had no effect. Cytochalasin B, cytochalasin D, and latrunculin B, all of which inhibit actin polymerization, had no effect on photopolarization, but arrested polar axis fixation. The role of calmodulin during polarization was investigated further. Calmodulin mRNA from the closely related brown alga Macrocystis pyrifera was cloned and the protein was expressed in bacteria. Photopolarization was enhanced following microinjections of this recombinant calmodulin into developing zygotes. Confocal imaging of fluorescein isothiocyanate-labeled recombinant calmodulin in photopolarized zygotes showed a homogenous signal distribution at 13 h AF, which localized to the presumptive rhizoid site at 15 h AF.     

Mechanisms, biology and inhibitors of deubiquitinating enzymes

The addition of ubiquitin (Ub) and ubiquitin-like (Ubl) modifiers to proteins serves to modulate function and is a key step in protein degradation, epigenetic modification and intracellular localization. Deubiquitinating enzymes and Ubl-specific proteases, the proteins responsible for the removal of Ub and Ubls, act as an additional level of control over the ubiquitin-proteasome system. Their conservation and widespread occurrence in eukaryotes, prokaryotes and viruses shows that these proteases constitute an essential class of enzymes. Here, we discuss how chemical tools, including activity-based probes and suicide inhibitors, have enabled (i) discovery of deubiquitinating enzymes, (ii) their functional profiling, crystallographic characterization and mechanistic classification and (iii) development of molecules for therapeutic purposes.