Occurrence of microbodies in chloromonadophycean algae

Microbody-like organelles occur in the cytoplasm of two chloromonadophycean algae,Vacuolaria virescens Cienkowsky andGonyostomum semen Diesing. Microbodies ofVacuolaria andGonyostomum have a granular matrix which lacks a crystalloid core; they are often present in close association with elements of the endoplasmic reticulum. The occurrence of microbodies in other algae is briefly reviewed.     

Centromeric organization in the chloromonadophycean alga Vacuolaria virescens

Observations with light and electron microscopes indicate the presence of a localized kinetochore in the phytoflagellate Vacuolaria virescens Cienk. (class Chloromonadophyceae). Thus, metaphase chromosomes have a clearly defined constriction which stains less densely than the remainder of the chromosome; at late metaphase the chromatids separate at definite points; acentric chromosome fragments may be formed. Ultrastructurally the kinetochore is recognizeable as a specialized area on the poleward surface of the chromosome into spindle microtubules are inserted.     

CHLOROPLAST STRUCTURE IN THE CHLOROMONADOPHYCEAN ALGA VACUOLARIA VIRESCENS1

The chloroplasts of Vacuolaria virescens Cienkowsky are present in large numbers between the cell membrane and the layer of cytoplasm surrounding the nucleus; they are disc-shaped structures ca. 3–4 μM long by 2–3 μM wide. Chloroplast bands consist of 3 opposed thylakoids with adjacent bands frequently interconnected. External to the girdle band is a chloroplast envelope separated from the cytoplasm by endoplasmic reticulum; there is no immediately apparent continuity between this endoplasmic reticulum and the nuclear envelope. Small electron-dense spheres in the chloroplast stroma are thought to be lipid food reserve. Eyespots and pyrenoids are absent.     

CHLOROPLAST PIGMENTS OF CHLOROMONADOPHYCEAE

The chloroplast pigments of 2 species of Chloro-monadophyceae have been examined. Both Gonyo-stomum semen and Vacuolaria virescens possessed only one chlorophyll, chlorophyll a. Gonyostomum contained 4 carotenoids, but not fucoxanthin and peridinin. Vacuolaria also possessed 4 carotenoids which have been identified as β-carotene and the 3 xanthophylls commonly found in the Xanthophyceae (as represented by Tribonema aequale). These are antheraxanthin, lutein monoepoxide, and an hydroxy lutein monoepoxide-like xanthophyll. The taxonomic significance of these findings, the first for the Chloromonadophyceae, is discussed.     

Chromosome Number and Morphology in Vacuolaria virescens (Chloromonadophyceae)

Vacuolaria virescens Cienk. has a chromosome number of 97±2.At metaphase these range in size between small dot-like chromosomes1 um in diameter to rod-shaped structures 12 µm long.Centromeres and less densely stained regions of chromosomesare recognizable.     

Competition between laboratory populations of green leafhoppers,Nephotettix spp. (Homoptera: Cicadellidae)

Intra- and interspecific competition between laboratory populations of four green leafhoppers, Nephotettix spp. was studied in the laboratory under three different temperature regimes of 24°C, 27°C and 30°C. For the single-species population of the three tropical species, the equilibrium density increased as the temperature increased. On the other hand, for the temperature species N. cincticeps, the highest equilibrium density was at the intermediate temperature and the lowest at high temperature. Interspecific interactions between two tropical (N. virescens vs. N. nigropictus), a tropical and a temperature (N. virescens vs. N. cincticeps) and a rice-feeding and a grass-inhabiting (N. virescens vs. N. malayanus) Nephotettix species were also studied in the laboratory at the three temperature regimes. Temperature differentially affected the outcome of competition between two Nephotettix species. Between N. virescens and N. nigropictus, the latter was more successful over the former at low and intermediate temperatures, while the former was more successful at high temperature. Between N. virescens and N. cincticeps, the temperate species inhibited the growth of the tropical species at low temperature while the tropical species inhibited the growth of the temperate species at high temperature. At intermediate temperature, the population of N. virescens persisted at a slightly higher density over the population of N. cincticeps. Between the rice-feeding N. virescens and the grass-inhabiting N. malayanus, regardless of temperature the population density of the latter was greatly reduced and later became extinct while the population of the former continued its growth. These consequences of competition between two Nephotettix species conformed fairly well to those predicted by theLotka-Volterra model using demographic parameters specified for each species.     

MITOSIS IN THE ALGA VACUOLARIA VIRESCENS

Details of mitosis in the chloromonadophycean alga Vacuolaria virescens Cienk. have been studied with the light microscope. The chromosomes are relatively large (up to μ in length at metaphase) and so mitotic stages are readily distinguishable. Chromosomes can be recognized in interphase nuclei as fine strands of chromatin. Contraction of these chromosomes marks the beginning of mitosis and continues progressively until the transition from metaphase to anaphase. Disintegration of nucleoli is complete by late prophase and nucleolar reformation begins in telophase. Some chromosomes exhibit less densely stained regions; centromeres are also present as indicated by their differential staining and by the behavior of chromosomes at metaphase and anaphase. At anaphase progeny chromosomes move apart parallel to the division axis of the nucleus. As anaphase progresses the chromosomes fuse at the polar surface of the progeny chromosome groups. This process continues in telophase and the chromosome groups become more spherical. By the end of telophase nucleolar reformation has begun and the chromosomes have relaxed to their interphase condition.     

Comparison of dispersal ability between the green leafhoppers,Nephotettix virescens andN. cincticeps (hemiptera: Cicadellidae) by tethered flight

Dispersal propensity of green leafhoppers was compared between a tropical species, Nephotettix virescens, and a temperate species, N. cincticeps. The flight ability was measured with tethered flight technique under laboratory conditions (25°C, 16L-8D). The pre-flight period was shorter and the flight duration was longer in N. virescens than in N. cincticeps in both sexes. No significant correlations were found between the flight activity and morphometric characters for either of the two species. The results suggested that dispersal propensity of N. virescens is higher than that of N. cincticeps.     

Vacuolaria tropicalis,a new species of Chloromonad from Southern Brazil

Vacuolaria tropicalis sp. nov. is here described as a new species of Chloromonadophyceae, observed in material collected during the summer of 1968–1969 from an artificial pool behind the greenhouses in the São Paulo Botanical Garden in the city of São Paulo. It is distinguished from the three previously described species of the genus chiefly by cell shape and by colour and localisation of the chromatophores.     

Contemporary evolution of a Lepidopteran species,Heliothis virescens,in response to modern agricultural practices

Adaptation to human‐induced environmental change has the potential to profoundly influence the genomic architecture of affected species. This is particularly true in agricultural ecosystems, where anthropogenic selection pressure is strong. Heliothis virescens primarily feeds on cotton in its larval stages, and US populations have been declining since the widespread planting of transgenic cotton, which endogenously expresses proteins derived from Bacillus thuringiensis (Bt). No physiological adaptation to Bt toxin has been found in the field, so adaptation in this altered environment could involve (i) shifts in host plant selection mechanisms to avoid cotton, (ii) changes in detoxification mechanisms required for cotton‐feeding vs. feeding on other hosts or (iii) loss of resistance to previously used management practices including insecticides. Here, we begin to address whether such changes occurred in H. virescens populations between 1997 and 2012, as Bt‐cotton cultivation spread through the agricultural landscape. For our study, we produced an H. virescens genome assembly and used this in concert with a ddRAD‐seq‐enabled genome scan to identify loci with significant allele frequency changes over the 15‐year period. Genetic changes at a previously described H. virescens insecticide target of selection were detectable in our genome scan and increased our confidence in this methodology. Additional loci were also detected as being under selection, and we quantified the selection strength required to elicit observed allele frequency changes at each locus. Potential contributions of genes near loci under selection to adaptive phenotypes in the H. virescens cotton system are discussed.     

Das sogenannte Alveolarplasma und die Schleimbildung beiVacuolaria virescens

Zusammenfassung FürVacuolaria virescens ist das Vorkommen, regelmäßig verteilter kugeliger Trichocysten in einer peripheren Plasmaschichte kennzeichnend. Auf äußere Reize hin werden sie ausgeschleudert und verquellen zu einer der Zelle anhaftenden Gallerte. Außerdem kann vermittels der pulsierenden Vakuolen Schleim erzeugt werden.Das Vakuolensystem besteht aus einer Hauptvakuole, die zunächst kegelförmig ist, sich dann abkugelt und schließlich in den Schlund entleert. Die Vakuolenbildung geht von einer zwischen der Hauptvakuole und dem abgeplatteten Zellkern hegenden Schichte zähen Plasmas aus, an deren oberem Rand zahlreiche Tröpfchen entstehen, die heranwachsen, zusammenfließen und sich schließlich zur Hauptvakuole vereinigen.In den Chromatophoren finden sich dünn scheibenförmige Körper; möglicherweise handelt es sich um Pyrenoide.Während der Drucklegung stieß ich auf eine Veröffentlichung von B.Fott [Mikroflora oravských raelin (Mikroflora der Orava-Moore), Preslia24, 1952], in der er das Vakuolensystem vonGonyostomum ovatum sp. n. genau beschreibt und angibt, daß andre Chloromonadinen, darunterVacuolaria virescens, sich wieGonyostomum verhalten. Meine Beobachtungen stimmen mit seinen im wesentlichen überein. Die von ihm in diesem Zusammenhang zitierten Abhandlungen vonKorikov, Hovasse undPoisson etHolland sind mir leider nicht zugänglich.     

SEXUAL REPRODUCTION AND GENETIC VARIATION IN THE SUBAERIAL ALGA CEPHALEUROS VIRESCENS (ULVOPHYCEAE,CHLOROPHYTA)

Cephaleuros virescens is a pantropical subaerial green alga with no known long‐range dispersal mechanisms. Sexual reproduction is relatively rare and may involve intragametangial fusion of identical, mitotically produced gametes. This situation may be a consequence of adaptation to the subaerial habitat. Genetic variation among populations of C. virescens may be very low and might be positively correlated to the distance (hence, time) separating populations. Thus, assessing the global biogeography of C. virescens requires analysis of what might be low levels of variation. Because C. virescens occurs on literally hundreds of different host species, the question of host‐races must also be considered. Preliminary analysis of local populations of C. virescens, originally obtained as field collections from three different host species and subsequently raised in culture, is the first step in addressing the biogeography of this alga. We are using the AFLP plant mapping protocol by PE Applied Biosystems to detect genetic variability in the three isolates of C. virescens. AFLP is a PCR‐based DNA fingerprinting technique that detects the presence or absence of restriction fragments rather than fragment length differences. Because the number of restriction fragments that can be detected with the AFLP technique is “virtually unlimited,” it is a very powerful tool for assessing the degree of relatedness or variability among cultivars or isolates. AFLP techniques have been used successfully to distinguish morphologically identical bacteria, determine relatedness among soybean accessions, reveal genetic variability within bee samples, and identifyfall armyworm strains and hybrids.     

Alkanes of four related moth species,Helicoverpa and Heliothis

Comparison of the presence and quantities of cuticular hydrocarbons has been used successfully for identifying sibling species and races of several groups of insects. This approach has been extended to four species of moths previously regarded as belonging to the same genus, Heliothis. Gas chromatography was used to quantify the numerous high-molecular weight alkanes found on the cuticle of two pairs of closely related species: Helicoverpa zea and Helicoverpa armigera, and Heliothis virescens and Heliothis subflexa. Both sexes of H. zea and H. armigera contained different quantities of several alkanes that could be used for unambiguous identification. Similar comparisons of H. subflexa and H. virescens showed four peak ratios that were different for each species. Sexual dimorphism was minor in H. subflexa and H. virescens.     

Communication interference in sympatrically occurring moth species

In moth species, females emit a species‐specific sex pheromone that is perceived over long distance by conspecific males. The species‐specificity in the chemical communication channel is achieved by a combination of unique components in specific ratios and sometimes also by interspecific behavioural antagonists to deter sympatrically occurring heterospecific males. In this study, we determined possible antagonistic effects in Helicoverpa gelotopoeon Dyar (Lepidoptera: Noctuidae) males to the major sex pheromone component of sympatrically occurring heliothine moths, Z11‐16:Ald, as well as to the sex pheromone of the sympatrically occurring Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae) (Z11‐16:Ald and Z9‐14:Ald). We also explored whether other co‐occurring species are attracted to these pheromone blends. Our field experiments showed that the addition of Z11‐16:Ald alone or in combination with Z9‐14:Ald inhibited trap catches of H. gelotopoeon males and that this inhibition depended on the concentration of these compounds. In addition, other moth species were attracted to the blends. Together, our results confirm the antagonistic effect of heterospecific sex pheromone compounds of H. virescens to H. gelotopoeon.     

Morphological features and chromosome numbers in cultures of five Cephaleuros species (Trentepohliaceae,Chlorophyta) from Japan

The morphological features and chromosome numbers were examined in cultures of five species of Cephaleuros; C. aucubae, C. biolophus, C. japonicus, C. microcellularis, and C. virescens, collected from Japan. On agar media of Bold's basal medium and CA medium, radial growth of the algal colonies was vigorous in C. virescens, moderate in C. aucubae, and poor in C. microcellularis, but varied from poor to moderate in isolates of C. biolophus and C. japonicus. Filaments of C. virescens branched at an angle less than 40° and just below the cross walls, while those of the other species branched at an angle greater than 40° without relation to the cross walls. The length of filamentous cells was significantly smaller in C. microcellularis than in the other species. The chromosome number differed with the species: n = 22 in C. aucubae, n = 34 in C. biolophus, n = 18 in C. japonicus, n = 12 in C. microcellularis, and n = 24 in C. virescens. The chromosome number did not vary between isolates from filaments and gametes in C. aucubae and C. biolophus and among those from filaments, gametes, and zoospores in C. japonicus and C. virescens.     

Discovery of novel trimethylalkanes in the internal hydrocarbons of developing pupae of Heliothis virescens and Helicoverpa zea

Novel trimethyl-branched alkanes which eluted with the monomethylalkanes were identified in the internal lipids of Helicoverpa zea but were not present in Heliothis virescens. Their structures were unique in that the first methyl branch occurred on carbon 2 and the 2nd and 3rd methyl branch points were separated by a single methylene. Novel trimethylalkanes identified from their chemical ionization and electron impact mass spectra were 2,18,20-trimethyltetratriacontane, 2,18,20-trimethylhexatriacontane, and 2,24,26-trimethyldotetracontane. Previous reports did not find these trimethylalkanes in the cuticular surface lipids of larvae, pupae or adults of either species. The internal pupal hydrocarbons of H. virescens and H. zea amounted to 123 μg and 304 μg per pupa, respectively. They consisted of n-alkanes (8 and 4%, respectively) and methyl-branched alkanes (88 and 94%, respectively). The n-alkanes ranged in chain length from approximately 21 to 35 carbons and the methyl-branched alkanes from approximately 26 to 55 carbons vs. methyl-branched alkanes from 28 to 37 carbons previously reported for hydrocarbons from the pupal cuticular surface. The major n-alkane was heptacosane (3.3 and 1.2%, respectively, in H. virescens and H. zea). The major methyl-branched alkanes in H. virescens were methylhentriacontane (15%), methyltritriacontane (12%) and dimethyltritriacontane (10%), and in H. zea were methylnonacosane (17%), dimethylnonacosane (9%) and methylhentriacontane (20%). Except for the novel trimethylalkanes, the methylalkane branch points were predominantly on odd-numbered carbons as has been reported for these and other species.     

Susceptibility of Helicoverpa zea and Heliothis virescens (Lepidoptera: Noctuidae) to Vip3A insecticidal protein expressed in VipCot™ cotton

Susceptibility of laboratory and field colonies of Helicoverpa zea (Boddie) and Heliothis virescens F. to Vip3A insecticidal protein was studied in diet incorporation and diet overlay assays from 2004 to 2008. Responses of field populations were compared to paired responses of University of Arkansas laboratory susceptible H. zea (LabZA) and H. virescens (LabVR) colonies. After 7 d of exposure, observations were made on number of dead larvae (M) and the number of larvae alive but remaining as first instars (L1). Regression estimates using M (LC₅₀) and M plus L1 (MIC₅₀) data were developed for laboratory and field populations. Susceptibility of laboratory and field populations exposed to Vip3A varied among different batches of protein used over the study period. Within the same batch of Vip3A protein, susceptibilities of laboratory colonies of both species (LabZA and LabVR) were similar. Field colonies were significantly more susceptible to Vip3A than the respective reference colonies of both species. Within field populations, susceptibility to Vip3A varied up to 75-fold in H. zea and 132-fold in H. virescens in LC₅₀ estimates. Variabilities in MIC₅₀s were up to 59- and 11-fold for H. zea and H. virescens, respectively.     

Responses of male Helicoverpa zea to single pulses of sex pheromone and behavioural antagonist

Male Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) flying in a pheromone plume respond to the loss of pheromone when they fly into a large pocket of clean air by going into crosswind casting flight in a mean of 0.48 s; 0.62 s after re‐contacting pheromone presented as a single pulse, they surge upwind in a kind of narrow zigzagging flight. After 0.36 s of surging, they lapse into casting flight once again in the clean air following the pulse. The addition of a known behavioural antagonist (Z)‐11‐hexadecenyl acetate (Z11–16:Ac), to the pheromone significantly increases the mean latency of the response to a single pulse to 0.85 s. No other aspects of the surge were significantly changed by the presence of antagonist in the single pulse of pheromone. Thus, unlike males of the related species, Heliothis virescens, which show significant changes in track and course angles when antagonist is present in single pulses, only an increased latency of response to a filament containing antagonist occurred in H. zea males. The increased latency could act cumulatively when the male is exposed rapidly and repeatedly to filaments in a natural plume and explain the profound arrestment effect of the antagonist in such plumes. The latencies to casting and surging in response to a pulse of pheromone blend are longer than those of the smaller species, H. virescens, and may be due to size‐related differences in manoeuverability of H. zea vs. H. virescens.