Phylogeny, diversity, and species delimitation of the North American Round-Nosed Minnows (Teleostei: Dionda), as inferred from mitochondrial and nuclear DNA sequences

Accurate delimitation of species is a critical first step in protecting biodiversity. Detection of distinct species is especially important for groups of organisms that inhabit sensitive environments subject to recent degradation, such as creeks, springs, and rivers in arid or semi-desert regions. The genus Dionda currently includes six recognized and described species of minnows that live in clear springs and spring-fed creeks of Texas, New Mexico (USA), and northern Mexico, but the boundaries, delimitation, and characterization of species in this genus have not been examined rigorously. The habitats of some of the species in this genus are rapidly deteriorating, and many local populations of Dionda have been extirpated. Considering the increasing concerns over degradation of their habitat, and pending a more detailed morphological revision of the genus, we undertook a molecular survey based on four DNA regions to examine variation over the range of the genus, test species boundaries, and infer phylogenetic relationships within Dionda. Based on analyses of two mitochondrial (cytb and D-loop) and two nuclear (Rag1 and S7) DNA regions from specimens collected throughout the range of Dionda, we identified 12 distinct species in the genus. Formerly synonymized names are available for two of these species, and four other species remain undescribed. We also redefine the known range of six species. The limited distribution of several of the species, coupled with widespread habitat degradation, suggests that many of the species in this genus should be targets for conservation and recovery efforts.     

Ultrastructure of Amoebophrya sp. and its changes during the course of infection

Amoebophrya is a syndinian parasite that kills harmful bloom forming algae. Previously uncharacterized ultrastructural aspects of infection and development were elucidated. The biflagellate dinospore has two mitochondria, electron-dense bodies, striated strips, trichocysts, and a nucleus with peripherally condensed chromatin. After finding an Akashiwo sanguinea host and adhering to its surface, the parasite penetrates the host surface, apparently using a microfilament based motility and electron-dense bodies within a microtubular basket in the process of parasitophorous vacuole membrane formation. After entering the host nucleus, possibly by a similar mechanism used to enter the host cell, the parasite cytosol expanded substantially prior to mitosis. From 12-36 hours mitochondria were inconspicuous but present. Chromatin condensation was variable. By 36 hours post-infection, parasites had multiple nuclei, a microtubule-supported cytopharynx, and were beginning to form a fully internal mastigocoel. By 48 hours, the characteristic "beehive" appearance was apparent with flagella projecting into a fully developed mastigocoel. The cytoplasm contained trichocysts, elongated mitochondria, and nuclei with peripherally condensed chromatin. Although Amoebophrya lacks an apical complex, its electron-dense bodies show functional similarities to apicomplexan rhoptries. Its lack of permanently condensed chromosomes, but compact dinospore chromatin, supports the idea that dinoflagellate permanently condensed chromosomes may be a remnant of a parasitic ancestor with a compact dispersal stage.     

四川及其周边地区野生草莓资源调查、收集与评价

调查了四川及其周边地区(未包括青海省)野生草莓资源种类和地理分布,将收集的野生草莓资源进行迁地保存,并对其农艺性状、抗非生物和生物胁迫能力的变异进行了主成分分析。结果表明,四川及其邻近周边高海拔地区野生草莓主要分布在26°14’～33°22’N、96°57’～108°13’E、海拔823～4531m的垂直空间范围内。黄毛草莓(Fragaria nilgerrensisSchlecht.)、五叶草莓(F.pentaphylla Lozinsk.)、西南草莓(F.moupinensis(Franch.)Card.)和纤细草莓(F.gracilis Lozinsk.)是该区域分布的4个野生草莓种类,其中五叶草莓有红果和白果两个类型。黄毛草莓浆果具有特殊的蜜桃香气。株高、叶柄长度、小叶长度、小叶宽度、茸毛、耐旱性和耐热性是野生草莓遗传性状的主要指标,茸毛数量与抗逆性密切相关。     

Coils in the membrane core are conserved and functionally important

With the increasing number of available α-helical transmembrane (TM) protein structures, the traditional picture of membrane proteins has been challenged. For example, reentrant regions, which enter and exit the membrane at the same side, and interface helices, which lie parallel with the membrane in the membrane-water interface, are common. Furthermore, TM helices are frequently kinked, and their length and tilt angle vary. Here, we systematically analyze 7% of all residues within the deep membrane core that are in coil state. These coils can be found in TM-helix kinks as major breaks in TM helices and as parts of reentrant regions.Coil residues are significantly more conserved than other residues. Due to the polar character of the coil backbone, they are either buried or located near aqueous channels. Coil residues are frequently found within channels and transporters, where they introduce the flexibility and polarity required for transport across the membrane. Therefore, we believe that coil residues in the membrane core, while constituting a structural anomaly, are essential for the function of proteins.     

Multi-scale assessment of macroinvertebrate richness and composition in Mediterranean-climate rivers

1. Similar constraints in distant, but climatically comparable, regions may be expected to yield biotic assemblages with similar attributes. Environmental factors that constrain communities at smaller scales, however, may be different between climatically similar regions. Thus, patterns observed at large scales may differ from those detected at small scales, and international comparisons should be focussed at multiple scales. 2. Mediterranean‐climate regions (MCRs) are characterized by remarkable seasonal variability in precipitation and temperature. Accordingly, rivers in these regions have seasonal and predictable floods and droughts, and temporary reaches are frequent. Present in six geographical regions of the world, MCRs have similar environmental constraints and are ideal for testing intercontinental similarities between macroinvertebrate communities. 3. We examined aquatic macroinvertebrate taxon richness and composition in MCRs at three scales: regional, reach and macrohabitat. At the regional scale, the Mediterranean Basin had the highest taxon richness at family level, and southwestern Australia the lowest. Taxonomic composition showed c. 85% similarity between the northern hemisphere MCRs of California and the Mediterranean Basin, which were followed in similarity by South Africa. The two Australian MCRs (South west and South) showed a similarity to each other of about 70% whereas the Chilean fauna was the most distinct. 4. At the reach scale, taxon richness was not significantly different between permanent and temporary reaches in any MCR, whereas taxonomic composition was significantly different among northern hemisphere MCRs. At the macrohabitat scale, taxon richness was not significantly different between lotic and lentic macrohabitats within any of the MCRs, but differences in macroinvertebrate communities were found between macrohabitats when considering regions. 5. Our results show that the strength of similarity between distant but climatically similar regions is scale‐dependent, being highest at the macrohabitat scale. Although the similarities in richness and composition at the macrohabitat scale are presumed to be universal, the seasonal predictability of drought in MCRs is expected to result in characteristic macroinvertebrate responses at the reach scale. We suggest, however, that regional evolutionary history and environmental characteristics may override this general pattern of a similar response of MCRs at different scales. The Mediterranean Basin and California, having similar historical and environmental condition, thus appeared as the most similar MCRs at all scales.     

Isolation and characterization of cold-regulated transcriptional activator <Emphasis Type="Italic">LpCBF3</Emphasis> gene from perennial ryegrass (<Emphasis Type="Italic">Lolium perenne</Emphasis> L.)

In plants, low temperatures can activate the CBF cold response pathway playing a prominent role in cold acclimation by triggering a set of cold-related gene expressions. CBF homologous gene, designated as LpCBF3, from a cold-tolerant perennial ryegrass (Lolium perenne L.) accession was identified. It carries the sequences for nuclear localization signal (NLS), AP2 DNA-binding domains and an acidic activation present in most of the plant CBF proteins. Southern analysis indicated the presence of three homologs of LpCBF3 gene in perennial ryegrass genome, and only one amino acid variation in LpCBF3 protein between cold-tolerant and -sensitive perennial ryegrass accessions. In their putative promoter regions, some differential regions were found. Northern blotting and RT-PCR analysis found that LpCBF3 reached the highest expression after 1.5 h of cold treatment (4 degrees C). The COR homologous gene, a downstream gene of CBF, can be expressed in the plant stem of cold-tolerant perennial ryegrass accessions without cold treatment. Without cold treatment, the COR gene cannot be activated in cold-sensitive perennial ryegrass accessions. Cold treatment can prompt expression levels of COR homologous genes in both perennial ryegrass accessions. In transgenic Arabidopsis, the overexpression of LpCBF3 with the 35S promoter resulted in dwarf-like plants, later flowering and greater freezing tolerance.     

The complete mitogenome of the Chinese bush cricket, Gampsocleis gratiosa （Orthoptera： Tettigonioidea）

The complete mitochondrial genome (mitogenome) of Gampsocleis gratiosa was determined. The 15, 929 bp in the size of G. gratiosa mitogenome contains a typical gene content, base composition, and codon usage found in metazoan. All 13 protein coding genes (PCGs) of the G. gratiosa mitogenome start with a typical ATN codon. The usual termination codons (TAA and TAG) were found from 10 PCGs. However, the atp6, nad4, and nad5 had incomplete termination codon (T). The anticodons of all tRNAs are identical to those observed in Drosophila yakuba and Locusta migratoria, and can be folded in the form of a typical clover leaf structure except for trnS (AGN). The secondary structure of trnS (AGN) was drawn according with the Steinberg-Cedergren tertiary structure. The A T content (67.4%) of the A T-rich region is relatively lower among the mitogenome regions, in contrast, it usually contains the highest A T content for most insects. Two isolated sequence repeat regions (202 bp) were found in the A T-rich region with mapping and secondary structure information.     

Molecular characterization of reciprocal crosses of Aerides vandarum and Vanda stangeana (Orchidaceae) at the protocorm stage

Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.     

Multiple sex chromosome system of X1X1X2X2/X1X2)Y type in lutjanid fish, Lutjanus quinquelineatus (Perciformes)

The karyotype and other chromosomal markers as revealed by C-banding and Ag-staining were studied in Lutjanus quinquelineatus and L. kasmira (Lutjanidae, Perciformes). While in latter species, the karyotype was invariably composed of 48 acrocentric chromosomes in both sexes, in L. quinquelineatus the female karyotype had exclusively 48 acrocentric chromosomes (2n = 48) but that of the male consisted of one large metacentric and 46 acrocentric chromosomes (2n = 47). The chromosomes in the first meiotic division in males showed 22 bivalents and one trivalent, which was formed by an end-to-end association and a chiasmatic association. Multiple sex chromosome system of X₁X₁X₂X₂/X₁X₂Y type resulting from single Robertsonian fusion between the original Y chromosome and an autosome was hypothesized to produce neo-Y sex chromosome. The multiple sex chromosome system of L. quinquelineatus appears to be at the early stage of the differentiation. The positive C-banded heterochromatin was situated exclusively in centromeric regions of all chromosomes in both species. Similarly, nucleolus organizer region sites were identified in the pericentromeric region of one middle-sized pair of chromosomes in both species. The cellular DNA contents were the same (3.3 pg) between the sexes and among this species and related species.