Jasmonic acid treatment and mammalian herbivory differentially affect chemical defenses and growth of wild mustard (Brassica kaber)

Summary. Jasmonic acid (JA) is a wound-related hormone found in most plants that, when applied exogenously, can induce increases in levels of chemical defenses in patterns similar to those induced by mechanical damage or insect feeding. Relative to responses to insect and pathogen attack, chemical responses of herbaceous plants to mammalian herbivore attack have been little studied. In a field experiment, we compared the effects of JA treatment and naturally occurring mammalian herbivory on the expression of trypsin inhibitors, glucosinolates, peroxidase activity and growth of wild mustard (Brassica kaber). Exogenous JA significantly increased trypsin inhibitor activity and glucosinolate concentration, and moderately increased peroxidase activity in the eighth true leaves of five-week-old plants, relative to untreated controls. In contrast, levels of these chemical defenses in the eighth true leaves or in regrowth foliage of plants that had ∼80% of their leaf area removed by groundhogs (Marmota monax) did not differ from that in undamaged and untreated controls. Although exogenous JA significantly elevated levels of chemical defenses, it did not affect height of plants through the season and only slightly reduced time to first flower. Groundhog herbivory significantly reduced height and delayed or abolished flowering, but these effects were not substantial unless coupled with apical meristem removal. We hypothesize that the lack of effect of groundhog herbivory on chemical defenses may be due in part to the speed and pattern of leaf area removal by groundhogs, or physiological constraints caused by leaf area loss. Despite having no effect on chemical defense production, leaf area loss by groundhogs was more costly to growth and fitness than the effects of JA application in this study, but only substantially so if coupled with apical meristem removal. We suggest that in general, costs of defense production in plants are likely to be minimal when compared to the risk of losing large amounts of leaf area or primary meristematic tissue. Thus, if they are effective at deterring herbivory, the benefits of inducible defense production likely outweigh the costs in most cases. Received 20 December 2000; accepted 3 May 2001     

Environmental levels of sediment-associated tri-n-butyltin chloride (TBTCl) and ionic regulation in flounders during seawater adaptation

The effects of exposure to sediment-associated tri-n-butyltin chloride (TBTCl) were examined in the euryhaline European flounder, Platichthys flesus (L.). The effects were quantified by measuring the changes in sodium efflux; Na⁺/K⁺-ATPase activity; and the numbers, areas, and distribution of chloride cells in the gills of freshwater-adapted fish, following a rapid transfer to seawater. Following the transfer to seawater, the Na⁺/K⁺-ATPase activity and the sodium efflux were significantly increased in the control group but remained unchanged in the TBTCl-exposed group. The normal morphological changes to the gill epithelium associated with seawater adaptation, which involve chloride cell distribution, took place in the control group but were significantly inhibited or delayed in the TBTCl group. The results presented in this study lead to the conclusion that environmental concentrations of tri-n-butyltin chloride in sediments are capable of significantly disrupting both the physiological and the morphological components of iono-regulatory functions of an estuarine flatfish.     

Chromosomal and nuclear characteristics of deep-sea hydrothermal-vent organisms: correlates of increased growth rate

A range of tissue and cell types from adult and juvenile stages of vent- and non-vent-dwelling deep-sea and shallow-water organisms were compared for signs of cell division, in preparation for a cytogenetic study of the different groups. Virtually all the Mid-Atlantic Ridge (MAR) vent species (bresiliid shrimp, bathymodiolid mussel, branchipolynoid polychaete, and a range of small gastropods) showed an abundance of metaphase chromosome spreads, indicating a generally high intrinsic rate of cell division, irrespective of their phylogenetic group. In comparison, similar tissues and life-history stages (i.e. gonad, developing eggs and gill) from two deep-sea bathypelagic crustaceans, Hymenodora and Gnathophausia, failed to yield any convincing evidence of cell division. This indicated that rates of growth and reproduction in the rest of the deep sea are extremely low. While much has been written about the contrasting growth rates of vent versus non-vent organisms, this is the first direct evidence of differences in cell-division rate between vent and non-vent species, which undoubtedly reflects contrasting conditions relating to food availability and temporal stability in these two deep-sea environments. The cells of shallow-water marine invertebrates (Mytilus, Nucella, Littorina, Pomatoceros and Crangon), based on the results of previous cytogenetic investigations, occupy an intermediate position with respect to their cell-division kinetics. When chromosome numbers, DNA content and nuclear diameters were compared between vent and non-vent species, this showed that no change had occurred in these characters since the time of vent colonisation. Representative chromosome spreads from hydrothermal-vent organisms are shown here for the first time.     

Spatial and temporal pattern in seagrass community composition and productivity in south Florida

We document the distribution and abundance of seagrasses, as well as the intra-annual temporal patterns in the abundance of seagrasses and the productivity of the nearshore dominant seagrass (Thalassia testudinum) in the south Florida region. At least one species of seagrass was present at 80.8% of 874 randomly chosen mapping sites, delimiting 12,800 km² of seagrass beds in the 17,000-km² survey area. Halophila decipiens had the greatest range in the study area; it was found to occur over 7,500 km². The range of T. testudinum was almost as extensive (6,400 km²), followed by Syringodium filiforme (4,400 km²), Halodule wrightii (3,000 km²) and Halophila engelmanni (50 km²). The seasonal maxima of standing crop was about 32% higher than the yearly mean. The productivity of T. testudinum was both temporally and spatially variable. Yearly mean areal productivity averaged 0.70 g m⁻²day⁻¹, with a range of 0.05–3.29 g m⁻² day⁻¹. Specific productivity ranged between 3.2 and 34.2 mg g⁻¹ day⁻¹, with a mean of 18.3 mg g⁻¹ day⁻¹. Annual peaks in specific productivity occurred in August, and minima in February. Integrating the standing crop for the study area gives an estimate of 1.4 × 1011 g T. testudinum and 3.6 × 10¹⁰ g S. filiforme, which translate to a yearly production of 9.4 × 10¹¹ g T. testudinum leaves and 2.4 × 10¹¹ g S. filiforme leaves. We assessed the efficacy of rapid visual surveys for estimating abundance of seagrasses in south Florida by comparing these results to measures of leaf biomass for T. testudinum and S. filiforme. Our rapid visual surveys proved useful for quantifying seagrass abundance, and the data presented in this paper serve as a benchmark against which future change in the system can be quantified. Received: 30 January 2000 / Accepted: 24 July 2000     

Chromosomal evolution in Mugilidae: karyotype characterization of Liza saliens and comparative localization of major and minor ribosomal genes in the six Mediterranean mullets

This study continues a comparative cytogenetic analysis of the fish family Mugilidae, reporting the karyotype characterization of the leaping mullet, Liza saliens, by C-banding, Ag- and fluorochrome-staining, and completing the fluorescence in situ hybridization (FISH)-mapping of the 18S and 5S rRNA genes (rDNA) to the chromosomes of the six Mediterranean mullets, namely L. saliens, L. ramada, L. aurata, Mugil cephalus, Chelon labrosus and Oedalechilus labeo. In all species, except M. cephalus, the 5S rDNA sites were localized on a medium-sized acrocentric chromosome pair, which was considered homeologous in all of them. In L. saliens, an additional 5S rDNA site was detected in a location close to the one shown by major ribosomal genes in M. cephalus, i.e. the subtelomeric region of chromosome pair 1. The 5S rDNA site in M. cephalus is located on the smallest chromosome pair of the complement, which, on the other hand, though on a different position, bears 18S rDNA in all the species of Liza and Chelon examined. The heterochromatin composition and the major and minor ribosomal gene locations suggest that the karyotype of L. saliens (subgenus Protomugil) can be considered intermediate between the karyotype of the more primitive M. cephalus and those of the other Liza (subgenus Liza) species and of the representatives of the more derived genera Chelon and Oedalechilus.     

Differences in herbivore preferences, phlorotannin production, and nutritional quality between juvenile and adult tissues from marine brown algae

 Juvenile and adult marine organisms differ in their morphology, chemistry, physiology, behavior, and ecology. Because juvenile algae are thinner, smaller, and have more delicate tissues than adults, they are often assumed to be more susceptible to grazers. We examined within-species food preferences of four common generalist herbivores for juvenile and adult tissues of eight common brown algae in two-choice laboratory food-preference experiments. Our results showed that juvenile algae did not tend to be a preferred food of herbivores. Juvenile tissues were significantly preferred over adult tissues in only four of the 32 combinations of algae and herbivores tested. In 12 experiments, adult tissues were preferred over juvenile tissues, and no choice occurred in the remaining 16 experiments. When sea urchins exhibited a preference, it was always for adult tissues. The other three herbivores, an isopod and two snails, were more variable in their choices, sometimes preferring juveniles, sometimes adults, and sometimes having no preference. We measured nitrogen and phlorotannin concentrations in adult and juvenile seaweeds to see whether these parameters were correlated with herbivore food preferences. Nitrogen levels were similar in juveniles and adults of three algal species and were higher in juveniles of two. Phlorotannin levels were higher in juveniles of four species and lower in juveniles of one. The other three species showed no differences in phlorotannin levels. Phlorotannin concentrations decreased with increasing juvenile size in three species and increased with increasing size in one species. Neither nitrogen nor phlorotannin concentrations explained overall herbivore food preferences for algae of different stages. Our results suggest that preferences of certain grazers for juvenile algae are not as strong as previously assumed and are dependent on herbivore species. Preferences between juveniles and adults are likely to be determined by a combination of morphological and chemical features of the tissues and the unique responses of herbivore species to those features. Received: 10 April 2000 / Accepted: 19 November 2000     

Molecular response of the sponge Suberites domuncula to bacterial infection

The aim of this study was the documentation of the molecular immune response of Suberites domuncula upon bacterial infection. Additionally, the bacteria that are naturally present in the sponge after prolonged aquarium maintenance were characterized. After 6 months of maintenance of S. domuncula in seawater aquaria, only one bacterial 16S rDNA sequence could be recovered, which belongs to the genus Pseudomonas. Concomitantly, morphologically uniform bacteria were found encapsulated in bacteriocytes. These findings indicate that certain bacteria, possibly of the genus Pseudomonas, are able to persist for long periods in host bacteriocytes. Subsequent to performing a previously established infection assay with S. domuncula, a potentially pathogenic Vibrio sp. was isolated from the tissues. Furthermore, the host tissue disintegrated and asexual propagation bodies (gemmules) were formed. In order to gain insights into the molecular events occurring after bacterial infection, the stress-response kinases, p38 protein kinase and JNK protein kinase, were analyzed. It is demonstrated that these two kinases are activated (phosphorylated) upon incubation of the tissue with the bacterial endotoxin lipopolysaccharide (LPS). Moreover, LPS strongly inhibits protein synthesis. It is concluded that there are many functionally different interactions between S. domuncula and bacteria and that the animal possesses mechanisms to differentiate between bacteria and to respond accordingly.     

Dimethylsulfoniopropionate and dimethylsulfide in the marine flatworm Convoluta roscoffensis and its algal symbiont

 A distinct smell of dimethylsulfide (DMS) was noted at the edge of the intertidal mudflat of Marennes-Oléron Bay, at the French Atlantic coast, where dense populations of the marine flatworm Convoluta roscoffensis Graff (Platyhelminthes: Turbellaria) were present. DMS is the cleavage product of dimethylsulfoniopropionate (DMSP). DMSP was shown to be present in high amounts in sediment containing the flatworm as well as in axenic cultures of the symbiotic alga Tetraselmis sp. that was isolated from the flatworm. In untreated sediment samples containing C. roscoffensis the concentration of DMS was as high as ∼55 μmol l⁻¹ sediment, and in samples that were fixed with glutaraldehyde the concentration of DMS was even three orders of magnitude higher (∼66 mmol l⁻¹ sediment). This rapid cleavage of DMSP to DMS in fixed samples was unexpected. Pure DMSP was stable in glutaraldehyde, and it was therefore concluded that a DMSP-lyase was responsible for cleavage in the field samples. The isolated symbiotic alga, Tetraselmis sp., did not show DMSP-lyase activity, indicating that DMSP-lyase may have been present in the flatworm, although the role of bacteria could not be excluded. The Chl a-specific DMSP content of C. roscoffensis (∼200 mmol g⁻¹) was much higher than that of Tetraselmis sp. (∼30 mmol g⁻¹). Possibly, DMSP was not only present in the symbiotic alga, but was also incorporated in the body tissue of the flatworm. It remains unclear what the function of DMSP is in C. roscoffensis. In Tetraselmis sp., but not in C. roscoffensis, DMSP increased with increasing salinity. It was concluded that salinity probably does not play an important role in the dynamics of DMSP and DMS in sediment containing C. roscoffensis. Received: 21 January 2000 / Accepted: 29 August 2000     

A Critique of the Recovery of Greenback Cutthroat Trout

Abstract: There are no examples of recovery of fish listed under the U.S. Endangered Species Act, but the number of federally threatened greenback cutthroat trout (  Oncorhynchus clarki stomias ) populations is approaching the delisting goal. We evaluated recovery of this subspecies in light of developing theory in conservation biology and with regard to recovery of other salmonids in the inland western United States. Four of the five criteria used to define populations that would count toward delisting appeared to underestimate the risk of extinction of those populations. Typically, recovery goals for numbers of greenback cutthroat trout populations were less stringent than those for other inland salmonids petitioned for listing or listed as threatened under the Endangered Species Act and were comparable to those for a federally endangered species. Before delisting is considered, we propose that historical populations be replicated in additional waters to protect genetic diversity and that existing populations be enlarged to reduce their vulnerability to demographic variation, to increase their access to refugia, and to permit reestablishment of mobile life histories. Existing stocks should also be evaluated to determine whether they represent distinct population segments.     

Ectomised conductors in the golden orb-web spider, Nephila plumipes (Araneoidea): a male adaptation to sexual conflict?

Male genitalia may facilitate sperm protection by acting as a plug that prevents or hinders future matings. The pedipalps (intromittant organs) of males of the orb-web spider, Nephila plumipes, have a conductor with a peculiarly curved ending and a triangular process near the terminal end. The tip of the conductor, including the process, breaks during most matings and remains inside the female genital tract. We explored the possible function of the conductor as a mating plug using the double-mating sterile-male technique. Our data are not consistent with a plug function because males use only one pedipalp in each mating, thus leaving an unobstructed insemination duct available for future matings; conductors of males mating with virgin females are not more likely to break than those of males mating with mated females, and second males show no preference for used or unused spermathecae. In addition, males that inserted their palp in the insemination duct that contained a tip of the conductor from a previous male obtained a share in the paternity of the female's clutch of eggs. Interestingly, the conductor is more likely to break if it is inserted in an unused spermatheca. We argue that several lines of evidence suggest that the conductor breaks as a result of intersexual conflict over the duration of copulation.