SEXUAL DIFFERENTIATION OF GRIFFITHSIA (CERAMIALES, RHODOPHYTA): NUCLEAR PLOIDY LEVEL OF MIXED-PHASE PLANTS IN G. JAPONICA1

Mixed-phase plants of Griffithsia japonica Okamura spontaneously occurred in a laboratory culture. Four female plants produced tetrasporangia and spermatangia in addition to their normal female reproductive structures (bisexual/mixed-phase plants), and four male plants produced tetrasporangia as well as spermatangia (male/mixed-phase plants). To determine the nuclear ploidy level of these mixed-phase plants, relative nuclear sizes of male, female, tetrasporangial, and mixed-phase plants were measured using a microscopic image analysis system. Haploid gametophytes could be distinguished from diploid tetrasporophytes by relative nuclear sizes, with the later having nuclei twice the size of the former. Relative nuclear sizes of the mixed-phase plants were similar to those of the haploid plants. Thus, the mixed-phase plants were determined to be haploid. Haploid mixed-phase plants of G. japonica have a potential to produce male, female and tetrasporangial reproductive structures. Sex determination models are discussed to explain "haploid" mixed-phase phenomena in red algae .     

Taxonomic revision of shrews of the genus Crociduras from the Sunda Shelf and Sulawesi with description of two new species (Mammalia: Soricidae)

The skulls of 387 shrews of the genus Crocidura sampled in peninsular Malaysia, Sumatra, Borneo, Java and Sulawesi were submitted to principal component and stepwise discriminant analyses. These analyses helped to delineate morphological taxa in this species-rich genus of mammals. Most morphologic groups could be attributed to described species, except one taxon from Sumatra and one from Sulawesi, which are described and named as new. Most of the 21 species recognized in this paper are endemic to one major island. Although Sulawesi has never been connected to the mainland, it supports at least six species, followed by Sumatra (5–6 species), Java and the Malay Peninsula (4 spp) and Borneo (3 spp). C. monticola is apparently the only widespread species whose distribution range covers the entire Malay Archipelago except the Philippines and Sulawesi. In contrast, the continental C. fuliginosa enters only marginally into the Sunda Shelf: its southernmost record is on the Malay Peninsula. This interpretation is completely different from the classical view that C. fuliginosa is a cosmopolitan species occupying the whole of Southeast Asia. Identification keys, tables of measurements and discriminant functions provided in this work may aid in identification of the various species and subspecies of Crocidura living in the Malaya Archipelago.     

Glinal variation in morphology among eastern populations of the waterstrider, Aquarius remigis Say (Hemiptera: Gerridae)

We examined morphological variation among populations of the stream-dwelling waterstrider, Aquarius remigis. A previous analysis of allozyme variation along a transect from southern Quebec to southern North Carolina revealed two distinct 'types' of A. remigis, north and south of a transition zone in southern New York and northern Pennsylvania. To assess the concordance of morphological and allozyme differentiation, adults were sampled from 15 populations along the original transect, as well as 11 additional populations in New York, Connecticut and New Jersey. Eight measurements of leg and body size were taken of preserved individuals. Univariate and multivariate analyses (multiple regression, PCA, discriminant analysis) reveal significant differentiation between northern and southern populations with a transition zone between latitudes 39^oand 42^onorth. Strong clines within each region and a clear transition between regions support previous suggestions that there are two distinct 'types' of this species in eastern North America. The transition zone, in morphology coincides with that revealed by the allozyme variation, but extends further south. The covariance of allozyme frequency and morphometric traits suggests that differentiation has occurred through vicariance and secondary contact, and that in this system, cladistic and adaptive differentiation are coupled.     

HMG-GoA reductase and terpenoid phytoalexins: Molecular specialization within a complex pathway

Terpenoid phytoalexins and other defense compounds play an important role in disease resistance in a variety of plant families but have been most widely studied in solanaceous species. The rate-limiting step in terpenoid phytoalexin production is mediated by 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), which catalyzes mevalonic acid synthesis. HMGRs are involved in the biosynthesis of a broad array of terpenoid compounds, and distinct isoforms of HMGR may be critical in directing the flux of pathway intermediates into specific end products. Plant HMGRs are encoded by a small gene family, and genomic or cDNA sequences encoding HMGR have been isolated from several plant species. In tomato, four genes encode HMGR; these genes are differentially activated during development and stress responses. One gene, hmg 2 , is activated in response to wounding and a variety of pathogenic agents suggesting a role in sesquiterpene phytoalexin biosynthesis. In contrast, expression patterns of tomato hmg l suggest a role in sterol biosynthesis and cell growth. Other plant species show an analogous separation of specific HMGR isoforms involved in growth and/or housekeeping function and inducible isoforms associated with biosynthesis of phytoalexins or other specialized "natural products". We are applying a variety of cell and molecular techniques to address whether subcellular localization and/or differential expression of these isoforms are key factors in determining end product accumulation during development and defense.     

Monitoring uptake and contents of Mg,Ca and K in Norway spruce as influenced by pH and Al,using microprobe analysis and stable isotope labelling

In a model system using intact spruce trees (Picea abies [L.] Karst.) we followed the path of magnesium, calcium and potassium during uptake into the root and during long-range transport into the shoot, by multiple stable isotope labelling. The roots of two- and three-year-old spruce trees originating from soil culture were removed from the soil and, in part or in toto, exposed to labelling solutions containing the stable isotopes ²⁵Mg or ²⁶Mg, ⁴¹K and ⁴²Ca or ⁴⁴Ca. Optical-emission-spectroscopy (ICP-OES) of plant fractions and labelling solutions was combined with the quantitative analysis of stable isotope ratios in sections of shock frozen, cryosubstituted material using the laser-microprobe-mass-analyser (LAMMA). This combination allowed us to distinguish, both in bulk samples and on the cellular level between (i) the fraction of elements originally present in the plant before the start of the labelling, (ii) the material taken up from the labelling solution into the plant and (iii) any material released by the plant into the labelling solution.In single-root labelling experiments, roots of three-year-old spruce trees, grown in nursery soil, were exposed to various pH conditions. The exchange of Mg and Ca with the labelling solution was nearly 100% in the cell walls of the mycorrhized finest roots. This exchange was only slightly affected by a step down to pH 3.5. The absolute Mg and Ca content in the cell walls was moderately reduced by incubation at pH 3.5 and strongly reduced in the presence of Al at this pH. After a pH 3.5 and 2 mM Al treatment we found Al in the xylem cell walls and the cortex cell lumina at elevated concentrations. To analyse the combined effect of high Al and high proton concentrations on the long-range transport, we used a split-root system. The root mass of an intact two-year-old spruce tree, grown in mineral soil, was divided into even parts and both halves incubated in solutions with two sets of different stable isotopes of Mg and Ca (side A: no Al, ²⁵Mg and ⁴²Ca; side B: +Al, ²⁶Mg and ⁴⁴Ca) and ⁴¹K on both sides. We observed a large uptake of Mg, Ca and K into the plant and a pronounced release. The net uptake of all three elements was lower from the Al-doted solution. In cross-sections of the apical shoot we found after seven-day labelling period about 60–70% of the Mg and Ca and 30% of the K content in the xylem cell walls originating from both labelling solutions. The clear majority of the Mg and Ca label originated from the Al-doted side.     

Relationships among microarthropods,fungi, and their environment

Temporal and spatial relationships in a maple-forest soil among mycophagous microarthropods, total hyphal length, vesicular-arbuscular mycorrhizal (VAM) fungus spores, microfungus diversity, root biomass and some abiotic variables (temperature, water content, pH, organic matter content) were investigated. Samples were obtained from spring 1991 to winter 1992 at four soil depths. Canonical correspondence analysis was used to analyze the data. Four species of sporulating VAM fungi were identified, along with 23 species of mites and springtails, 9 of which were common. Hyphal length, VAM fungus spores, and soil animals peaked in spring and autumn. Canonical correspondence analysis suggests that animal abundance and success in the soil is dependent on a number of environmental variables. The most important variables that influence microarthropod community structure are: (i) temperature, (ii) water content, (iii) pH, (iv) total length of fungal hyphae, and (v) diversity of darkly-pigmented fungi. However, the relative importance of these variables changes with increasing soil depth. We have also shown a relationship between arthropod populations and their food supply under field conditions, a phenomenon that has been demonstrated previously under controlled laboratory conditions.