Phenotypic plasticity in age and size at maturity and its effects on the integrated phenotypic expressions of life history traits of Cardamine flexuosa (Cruciferae)

We analyzed variation in phenotypic plasticity of life history traits between two Cardamine flexuosa populations based on differences in plasticity of age and size at maturity. C. flexuosa (Cruciferae) is a facultative, vernalization-sensitive, long-day annual, and its phenology and the phenotypic expressions of many life history traits are largely controlled by photoperiod and vernalization in natural populations. We used plants from two populations which differed in their responses to chilling and photoperiod treatments. The timing of developmental processes was changed by controlling temperature and photoperiod regimes in growth chambers. Plasticity in size at maturity was analyzed as changes in a growth trajectory using two parameters, age at maturity (Δt) and growth rate (k). Both traits showed plasticity, but differences between the populations were found mostly for Δt. Distinctive differences in size at maturity of individuals in the two populations were mainly due to different amounts of plasticity in Δt. Variations in plasticity of nine other life history traits and their associations to age and size at maturity were also analyzed. Variation for eight of the traits can be described, at least in part, as a function of age and size at maturity for both populations, and most of the variation in the total number of seeds was explained by age and size at maturity. Only age at maturity had any effect on changes in resource allocation. The nine life history traits were integrated through associated character expressions with age and size at maturity. Changes in the association between a trait and age and/or size at maturity were rather conservative compared to changes in the plasticity of a trait between the two populations. Associations with age and size at maturity are mostly explicable in terms of inherent relationships in the developmental processes, and they may limit the ecological range expansion and the adaptive evolution of plasticity in C. flexuosa. The negative correlation between reproductive allocation and age at maturity can be a cost of delaying maturation in C. flexuosa.     

A Novel Myosin-like Protein (Myocilin) Expressed in the Connecting Cilium of the Photoreceptor: Molecular Cloning, Tissue Expression, and Chromosomal Mapping

We have isolated a human cDNA clone encoding a novel acidic protein of MW 55,000 that we designated “myocilin” since it has homology to myosin and is localized preferentially in the ciliary rootlet and basal body of the connecting cilium of photoreceptor cells. The deduced amino acid sequence of human myocilin showed significant homologies with nonmuscle myosin ofDictyostelium discoideumin the N-terminal region and also with olfactomedin of bullfrog in the C-terminal region. Myocilin contained a leucine zipper-like motif similar to that seen in kinectin and other cytoskeletal proteins. These findings suggest that myocilin is a novel cytoskeletal protein involved in the morphogenesis of ciliated neuroepithelium such as photoreceptor cells. The myocilin gene (MYOC) was mapped to human chromosome 1q23–q24 by fluorescencein situhybridization.     

Genomic Organization and Complete Nucleotide Sequence of the HumanPWP2Gene on Chromosome 21

The humanPWP2gene is the human homologue of the yeast periodic tryptophan protein 2 (PWP2) gene and is a member of the gene family that contains tryptophan-aspartate (WD) repeats. Genomic sequencing revealed that the humanPWP2gene consists of 21 exons spanning approximately 24 kb and locates just between the two genes EHOC-1 and KNP-I and distal to aNotI site of LJ104 (D21S1460) on chromosome 21q22.3. Analysis of the 5′-flanking DNA sequence revealed that the upstream region of thePWP2gene is associated with a CpG island containing theNotI site of LJ104. SincePWP2is considered to be a candidate for genetic disorders mapped in the 21q22.3 region, the information including nucleotide sequence and genomic organization of thePWP2gene should be invaluable for the mutation analysis of the corresponding genetic disorders.     

Partial and complete adenine phosphoribosyltransferase deficiency associated with 2,8-dihydroxyadenine urolithiasis: kinetic and immunochemical properties of APRT

We have studied adenine phosphoribosyltransferase (APRT) in the hemolysates from the families of 2,8-dihydroxyadenine urolithiasis associated with partial deficiency of APRT (the Japanese type) and complete deficiency of APRT (the null type). The APRT in the control subjects was found to be heat-stable at the physiological concentration of phosphoribosylpyrophosphate (PRPP), which was close to the value of its Km for PRPP. The APRT in the Japanese type showed 10 times higher Km values for PRPP and needed a comparably increased level of PRPP for stability in vitro. No change in red cell PRPP was found in the Japanese type of APRT deficiency. The content of APRT enzyme protein was decreased in the hemolysates of the Japanese type, probably due to its lability at the level of PRPP present in the cells. The heterozygote of the null type also had labile enzyme molecules at the physiological PRPP concentration.     

Amino-terminal structure of spoOA protein and sequence homology with spoOF and spoOB proteins

Summary The previously reported nucleotide sequence of the spoOA coding region of Bacillus subtilis suggested that the protein is initiated with either of two possible initiation codons, ATG and GTG, 84 base pairs apart. To determine which codon is utilized as an initiator in B. subtilis, we constructed a fusion gene in which the promoter and NH₂-terminal region of the spoOA gene was connected to the chloramphenicol acetyltransferase gene (cat gene). After introduction of the plasmid carrying the spoOA-cat fusion gene into B. subtilis cells, the fusion protein was purified by affinity chromatography. The sequence of NH₂-terminal amino acids of the fusion protein was determined and the result established that the GTG codon is utilized as an initiator in B. subtilis.Comparison of the amino acid sequences revealed a marked homology between the spoOA (NH₂-terminal half) and spoOF proteins. A less striking but significant homology was also found between the spoOA (COOH-terminal half) and spoOB proteins. This suggests the presence of a common functional domain structure for these proteins that are supposed to play key regulatory roles in sporulation.     

Isolation of giant DNA fragments from flow-sorted human chromosomes

We have established a method using a conventional cell sorter equipped with a single argon laser to sort intact human chromosomes that can be used as a source for the production of giant DNA fragments. Various improvements were made to both the equipment and sorting method to enhance the sorting resolution and avoid destruction of chromosomal DNA. Using this improved method chromosomes 21 and 22 were sorted from the B-lymphoblastoid line GM00130B, digested with the rare cutting restriction endonuclease NotI, and analyzed by pulsed field gel electrophoresis followed by Southern hybridization using the Alu repetitive sequence as a probe. More than 25 discrete NotI giant DNA fragments ranging from 50 kb to longer than 2.5 Mb were separated and the size distribution pattern was unique for each chromosome, indicating successful sorting of intact chromosomes. The cumulative size of these Alu-positive NotI DNA fragments were 22.7 Mb and 25.5 Mb for chromosomes 21 and 22, respectively. These values are 47% and 49% of the estimated size of chromosomes 21 (48 Mb) and 22 (52 Mb).     

Possible ecological implications of floating microbial assemblages lifted from the lakebed on an Antarctic lake

Microbial assemblages can be found drifting/floating in lake water and being washed ashore in continental Antarctica. Two field studies in early and late January 2008 measured the light utilization properties and photosynthetic responses of these assemblages, which were then compared with those of pelagic and benthic microbial communities to evaluate the ecological implications of this phenomenon. The nutrient concentrations were low in the lake water, indicating oligotrophic conditions. Based on microscopic and pigment analysis, both the floating and benthic communities were mainly composed of Oedogonium sp. (Chlorophyceae), followed by cyanobacteria, diatoms, and dinoflagellates. Floating assemblages had a firmer and denser structure, and possessed more rich carotenoids than the benthic community. Measurements of photosynthesis conducted in early January indicated that the activities of the floating assemblages were considerably low. In late January almost all floating assemblages on the lakeshore turned white because of freezing and drying by the ambient temperature decrease, and had no photosynthetic signals. These results suggest that the floating assemblages could spontaneously lift off from the lakebed because of the bubbles created by photosynthesis and then repeatedly roll, flip, sink, or float depending on buoyancy. In addition, this phenomenon seemed to greatly change the cycling of matter by transporting the lake’s photosynthetic products to the surrounding ecosystems, then give the benthic subsurface communities in dark regions a chance to reactivate such as gap regeneration in the case of climax forest, and also allow the floating assemblages to restart photosynthesis at the top of the lakebed by resinking.     

Photophysiological responses of phytobenthic communities to the strong light and UV in Antarctic shallow lakes

Light environment, community structure, pigments, and photophysiological properties of mat-forming phytobenthos were studied in four shallow Antarctic lakes in 2007 at maximum water depths of 1.7–2.5 m. All lakes were oligotrophic, and water transparencies were high, enabling 45–60% of photosynthetically active radiation (PAR, 400–700 nm) and 20–40% of ultraviolet radiation (300–400 nm) to reach the lake beds. Phytobenthic mats were dominated by cyanobacteria and green algae. Little PAR_L (500–700 nm) penetrated through the firm mat in the shallowest lake, while in the other lakes more (>20%) PAR_L got through the mats to the subsurface mat layers. Photochemical activities indicated almost no photoinhibition but low photosynthetic efficiency in all mat surface layers. Non-photochemical quenching was rarely detected, suggesting excess energy dissipation may not be efficient in the UV-rich environment. There was a positive correlation between photo-protective substances and incident radiation in the mats, and an inverse correlation between such substances and photochemical efficiency, suggesting that the phytobenthos survive by changing a light-protection/utilization balance. The communities under strong UV-B and PAR had firm mat textures and were characterized by high UV/photo-protective substance ratios that make them less transparent. Maximum relative electron transportation rates (rETR_max) and photochemical efficiencies, however, were low, possibly because the protective substances prevent efficient light usage. In contrast, communities under mild light were characterized by lower substance ratios and softer textures, while rETR_max values and photochemical efficiencies were greater. The phytobenthic mat surface seems to act as a filter for strong and harmful light, typically penetrating through the clear water of Antarctic lakes, and produces a milder light environment for the subsurface mat organisms.