Changes in sperm tail development associated with Y chromosome meiotic drive leading to an excess of males in the medfly Ceratitis capitata (Diptera: Tephritidae)

The Mediterranean fruit fly Ceratitis capitata (Wied.) normally produces the sexes in equal ratio but strains carrying the Y chromosome meiotic drive MP (male‐producing) factor show an excess of males. This is associated with a loss of sperm, and abnormal sperm structure in terms of multiple axonemes, atypical numbers of mitochondrial derivatives, and sometimes an incorrect initial orientation of paracrystalline bodies to the axoneme. Sperms are bundled together within spermatocysts, and those with depleted content and abnormalities occur in the same MP testes as normal spermatocysts. The maximum number of sperms per cyst in control strains was 256, each with a single axoneme originating from a single centriole (kinetosome). The maximum per cyst in MP strains was also 256 but MP cysts contained up to 300 axonemes, providing evidence of multiplication of centrioles. The structural changes in MP sperm are discussed in relation to similar abnormalities reported in the mosquito Aedes aegypti inheriting the Y chromosome meiotic drive haplotype MD. The evolutionary significance of this phenomenon is considered. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 351–359.     

Regulatory T cells and control of the germinal centre response

Germinal centres (GCs) are specialised lymphoid microenvironments that form in secondary B-cell follicles upon exposure to T-dependent antigens. In the GC, clonal expansion, selection and differentiation of GC B cells result in the production of high-affinity plasma cells and memory B cells that provide protection against subsequent infection. The GC is carefully regulated to fulfil its critical role in defence against infection and to ensure that immunological tolerance is not broken in the process. The GC response can be controlled by a number of mechanisms, one of which is by forkhead box p3 expressing regulatory T (Treg) cells, a suppressive population of CD4⁺ T cells. A specialised subset of Treg cells – follicular regulatory T (Tfr) cells – form after immunisation and are able to access the GC, where they control the size and output of the response. Our knowledge of Treg cell control of the GC is expanding. In this review we will discuss recent advances in the field, with a particular emphasis on the differentiation and function of Tfr cells in the GC.     

Resistance in sorghum to shoot fly Atherigona soccata: Evidence for the source of leaf surface wetness

Leaf surface wetness (LSW) of the central whorl leaf of sorghum seedlings has been associated with susceptibility to shoot fly. Previous physical and physiological evidence suggested that LSW originates from the plant. This was confirmed by radioactive labelling methods using tritium and carbon-14. Tritiated water applied to the soil of potted seedlings was translocated to the surface of the whorl leaf. There were significant differences in the amount of tritiated water collected from susceptible (CSH 5) and resistant (IS 18551) genotypes. Studies with carbon-14 labelling of sorghum seedlings indicated the presence of (small amounts of) solutes in the surface water that may affect larval movement and survival.     

Interrogating accessibility of telomeric sequences with FRET-PAINT: evidence for length-dependent telomere compaction

Single-stranded telomeric overhangs are ∼200 nucleotides long and can form tandem G-quadruplex (GQ) structures, which reduce their accessibility to nucleases and proteins that activate DNA damage response. Whether these tandem GQs further stack to form compact superstructures, which may provide better protection for longer telomeres, is not known. We report single-molecule measurements where the accessibility of 24–144 nucleotide long human telomeric DNA molecules is interrogated by a short PNA molecule that is complementary to a single GGGTTA repeat, as implemented in the FRET-PAINT method. Binding of the PNA strand to available GGGTTA sequences results in discrete FRET bursts which were analyzed in terms of their dwell times, binding frequencies, and topographic distributions. The binding frequencies were greater for binding to intermediate regions of telomeric DNA compared to 3′- or 5′-ends, suggesting these regions are more accessible. Significantly, the binding frequency per telomeric repeat monotonically decreased with increasing telomere length. These results are consistent with telomeres forming more compact structures at longer lengths, reducing accessibility of these critical genomic sites.     

Precursors of Asparagine in Lupins

Fumaric acid-1,4-¹⁴C, L-aspartic acid-4-¹⁴C, and glycine-2-¹⁴Cwere supplied for 1 to 3 h to etiolated lupin shoots that wererapidly synthesizing asparagine. The distribution of ¹⁴C inaspartate and asparagine isolated from this material was determined.With aspartic acid-4-¹⁴C adminstration radioactivity was mostlyin carbons 1 and 4 with carbon 4 having the greater amount;with fumaric acid 1,4-¹⁴C administration, radioactivity wasagain mostly in carbons 1 and 4. Aspartate labelled by fumaratecontained relatively more radioactivity in carbons 2 and 3.Little radioactivity from glycine-2-¹⁴C entered either aspartateor asparagine and this was mainly in carbon 2 and 3. It is concludedthat asparagine synthesis form aspartate is a major pathwayof asparagine biosynthesis in lupins.     

Use of a model and toxicity data to predict the risks to some wild plant species from drift of four herbicides

The dose responses of 14 wild plant species (two grasses, two legumes, one annual and nine perennial dicotyledons), not usually recognised as weeds, to four herbicides (asulam, glyphosate, MCPA and mecoprop) were measured in glasshouse experiments. Glyphosate was the most toxic; seven of the species tested had EDu, values (measured as shoot dry weight) of < 1.0 μg/plant, compared with only one species for MCPA and mecoprop. Asulam was the least toxic. Results were used to indicate the risk to each species from drift damage. A model of spray drift, based on that developed by Thompson & Ley (1982) for evaporating droplets, was rescaled to allow for field application rates and used to predict the distances travelled by given doses of herbicide. This gave acceptable agreement with reports for drift damage in the field, and predicted that only glyphosate sprayed at the highest recommended concentration might be unsafe to some of the species examined. The two herbicides sometimes used as volatile formulations (MCPA and mecoprop) did not cause damage at the small doses likely to result from exposure to vapour in the field.     

The Functional and Evolutionary Significance of the Ultrastructure of the Ophryoscolecidae (Order Entodiniomorphida)1

ABSTRACT. Ophryoscolecid ciliates from the order Entodiniomorphida show a series of morphological types which have been interpreted as an evolutionary lineage. In this study, ultrastructural information from three species—the ancestral Entodinium, intermediate Eudiplodinium maggii, and the advanced Epidinium—has been evaluated in terms of ophryoscolecid evolution. The infraciliature, nuclei, contractile vacuoles, cortex, and cytoplasm are all very similar and sometimes indistinguishable among the three species, suggesting a close relationship. The cytoalimentary system, however, shows considerable interspecific variation. The cytopharynx differs in position and extent of microtubules and microfilaments, varying in appearance from a microtubular to a microfilament-based mechanism while retaining similar component structures. The esophagus, a zone of cytoplasm extending from the cytopharynx and delimited by a microtubular/fibrillar wall, is rudimentary in Entodinium, sac-like in Eudiplodinium, and tube-like in Epidinium, where it also has convoluted walls and a sheath of fibrous material that suggest an expansible-contractile structure. These variations have been related to type of food particle ingested. The capability of the cytoalimentary systems seems to be increased so that the more advanced forms can exploit a food resource, in the form of large plant fragments in the ruminal fluid, not available to the simpler, ancestral forms, which tend to ingest small particles such as bacteria and starch grains. The original evolutionary lineage based on morphological studies using light microscopy is supported by our observations, in these three forms, of ultrastructural variations in the cytopharynx and in their relationship with diet via possible ingestion mechanisms. Additional support for this evolutionary analysis comes from preliminary studies of other ophryoscolecids in which the cytoalimentary organization is consistent with their positions relative to one another in the evolutionary scheme.     

Isolation Conditions for High Yields of Protoplasts from Laminaria saccharina and L. digitata (Phaeophyceae)

In an investigation of the main factors determining protoplastyield in Laminaria saccharina and L. digitata, protoplasts wereisolated from epidermal, cortical and medullary cells of vegetativethallus by incubation with commercial cellulases, crude andpurified mannuronate lyases and purified guluronate lyases.Treatment of the tissue with the calcium chelator EGTA beforeenzymatic digestion greatly increased the protoplast yield.Preplasmolysis was also necessary to obtain large numbers ofhealthy protoplasts and this was most effective when carriedout during chelation with EGTA. Purification of the mannuronatelyases by ion exchange chromatography reduced the toxicity ofthe crude enzyme preparation. The activities of the wall degradingenzymes were differentially influenced by pH and the optimumfor alginate-lyase activity (8.0) was higher than that for cellulaseactivity (<6.0). Protoplast yield decreased linearly withincreasing pH in the enzyme medium over the range tested (6.0–8.0),and this suggests that cellulases are more critical to walldigestion than alginate-lyases. Ionic osmotica gave improvedyields compared with sugar alcohols or sugars. Increasing thecalcium concentration of the enzyme medium brought about anexponential decrease in protoplast yield and wall digestionwas almost completely inhibited at concentrations exceeding8.0 mol m^–3. However, low levels of calcium (<2.0 molm^–3) were beneficial to protoplast viability. Yields of10⁷ to 10⁸ protoplasts g^–1 fr. wt. were consistently obtainedand 20% to 30% of these regenerated new cell walls within 1–2d of culture. Key words: Laminaria protoplasts, cell wall, alginate-lyases     

Resistance of potatoes transgenic for a cry1Ac9 gene, to Phthorimaea operculella (Lepidoptera: Gelechiidae) over field seasons and between plant organs

Stable performance of insect‐resistant transgenic plants across field seasons and between plant organs damaged by the insect pest is critical for management of this resistance in the field. To evaluate this, potato (Solanum tuberosum) lines transgenic for a cry1Ac9 gene with resistance to potato tuber moth (Phthorimaea operculella) were established in the field during the southern hemisphere summers of 1997/98, 1998/99 and 1999/00 as small field plots, each of 10 plants. Replicate plots of the non‐transgenic parent cultivars (at least one for every three independently derived transgenic lines) were planted randomly throughout the trials. Field‐grown foliage was challenged with larvae in the laboratory and a growth index (GI) was calculated for recovered larvae from each transgenic and non‐transgenic potato line. Larval growth on young and mature leaves, and on newly harvested or stored tubers was also measured in the laboratory. Foliage from the transgenic lines inhibited larval growth in all seasons tested. For both control and transgenic lines, larvae had slightly lower GIs when reared on mature leaves compared with young leaves, although the correlation between mean GI for young and mature transgenic leaves was high (r = 0.97). The correlation between the mean GIs of larvae on newly harvested tubers and on those stored for 5 months was also high (r = 1.0). However, the GIs of larvae on newly harvested transgenic tubers were larger than on transgenic tubers stored for 5 months. The relative growth indices (RGI = mean GI/number days before final weighing) of larvae reared on newly harvested tubers from transgenic lines were generally higher than those from young transgenic foliage, while the RGIs of larvae reared on non‐transgenic tubers were slightly lower than those fed non‐transgenic foliage. The correlation between mean RGIs of larvae fed tubers or foliage was 0.62. The transgenic potato lines exhibited stable resistance to larvae across field seasons, between affected plant organs, and between plant organs of different ages.