Rhabdomeric membrane and smooth endoplasmic reticulum in photoreceptors of Manduca sexta: modulations associated with the diurnal light/dark cycle and effects of chromophore deprivation

Summary Aberrations of photoreceptor ultrastructure resulting from carotenoid/retinoid (vitamin A) deprivation were studied in the retina of Manduca sexta. The syndrome of chromophore deficiency included hypertrophy of smooth endoplasmic reticulum, variable dilation of rhabdomeric microvilli, the insertion of endomembrane fingers into such enlarged microvilli, and the formation of rhabdomeric vacuoles, intracellular compartments containing microvilli similar to those of the rhabdomere. Retinas were processed either with conventional procedures employing preliminary aldehyde fixation followed by heavy metal postfixation, or by fixation and incubation in unbuffered OsO₄. The latter method deposits osmium throughout the endomembrane system, within the rhabdomeric vacuoles, and in the extracellular space of the rhabdom. However, the intravillous fingers were rarely impregnated with osmium, despite their continuity with densely stained cisternae of the smooth endoplasmic reticulum. We suggest that the insertion of endomembrane fingers into dilated microvilli results from a cytoskeleton-mediated link between cisternae of the smooth endoplasmic reticulum and the rhabdomeric membrane, an association that may be important in the turnover of photoreceptor membrane. We interpret endomembrane hypertrophy and development of rhabdomeric vacuoles as symptoms of disturbance in the pathway leading to the assembly of the rhabdomere resulting from reduced synthesis of visual pigment.     

Pea lectin is correctly processed,stable and active in leaves of transgenic potato plants

A gene encoding the preproprotein of the pea (Pisum sativum) lectin was expressed in transgenic potato plants using a cauliflower mosaic virus (CaMV) 35S promoter or a tobacco ribulose bisphosphate carboxylase small subunit (ssRubisco) promoter. Presence of the pea lectin to levels greater than 1% of total soluble leaf protein was detected by radioimmunoassay (RIA). The pattern of expression derived from the two promoters was established using both RIA and a squash-blot immunolocalisation technique. Western blotting demonstrated that the preproprotein was correctly processed, generating and subunits that assembled to give an isolectin form observed in pea seeds and roots. It was also found that the haemagglutination activity and specificity of pea lectin synthesised in transgenic potato leaves was comparable to purified lectin from pea cotyledons.     

Lipolytic and peroxidative enzyme expressions in cultured potato tuber cells

Potato cells (cv. Norchip) were cultured from tuber parenchymal tissue and subcultured to dissociate and habituate the despecialized cells. After several subculturings on a minimal nutrient media, this line of cells demonstrated repeatable physical growth profiles for dry weight (DW), fresh weight (FW) and protein. Two enzymes of plant lipid metabolism were investigated, lipolytic acyl hydrolase (LAH) and lipoxygenase (LOX), which respectively liberate and peroxidize fatty acids from lipid in cellular membranes. LAH, measured as p-nitrophenyl palmitate hydrolase, was present in this line of cells in easily detectable amounts (317 units g^-1 DW) albeit much lower than that found in mother tuber (9878 units g^-1 DW). The presence of LAH in this line is significant because LAH isozymes are often described as storage proteins, yet activity per gram fresh weight in these unorganized cells is reasonably constant until culture growth exits the linear phase. However, LOX, the most active free fatty acid metabolizing enzyme in potato tubers (89,800 units g^-1 DW), was not detectable in this line of callus or suspension cultured cells. The absence of LOX activity in this line of cells was verified by a number of assay approaches and was confirmed by activity staining of extracted enzymes separated in polyacrylamide gels. The absence of LOX in these cultured cells is especially important in determining the functions of this lipid peroxidation system and how it may be genetically regulated.Mention of company or trade name does not imply endorsement by the United States Department of Agriculture over others not named.A laboratory cooperatively operated by the Midwest Area, Agricultural Research Service, U.S. Department of Agriculture, The Minnesota Agricultural Experiment Station, the North Dakota Agrcultural Experiment Station, and the Red River Valley Potato Grower's Association.     

A detailed study of the regulation and evolution of the two classes of patatin genes in Solanum tuberosum L.

The class-specific expression of patatin genes was investigated by analysing four new patatin genes. A class I patatin gene from cv. Berolina as well as a class I and two class II patatin genes from the monohaploid cultivar AM 80/5793 were isolated and partially sequenced. Sequence comparison indicates rearrangements as the major source for the generation of diversity between the different members of the classes. The expression of single genes was studied in potato plants transformed with chimaeric genes where the putative patatin promoters were fused to the GUS reporter gene. A detailed histochemical analysis reveals that both class I genes are expressed as the previously described class I patatin gene B33 from cv. Berolina [1], i.e. in the starch-containing cells of potato tubers and in sucrose-induced leaves. The class II gene pgT12 shows the same pattern as the previously described class II gene pgT2 [2], i.e. expression in root tips and in the vascular tissue of tubers, whereas no activity was detectable for pgT4. Thus the expression pattern of both classes of genes seems to be stable at least within or even between different cultivars.     

Temporal events of gypsy moth vitellogenesis and ovarian development

Abstract The vitellogenic period of gypsy moth ovarian development starts on day 3 of the pupal stage and continues through adulthood. During this period, rapid increases occur in follicle size, protein content, and wet weight of the ovary. Patency is observed on day 3 of the pupal stage.
Pre-vitellogenic follicles are formed in the last larval stadium. Newly formed follicles detach from the germarium on day 4, and increase rapidly to 140 per ovariole at the end of the last larval stadium. The pre-vitellogenic follicles are uniformly around 50 um in diameter. No vitellogenin is incorporated into the oocytes until the pupal stage.
Polyacrylamide gel electrophosesis (PAGE) in the presence of sodium dodecylsulphate (SDS) analysis of male and female haemolymph samples and vitellogenic ovaries demonstrates the presence of two female-specific subunits of vitellogenin of 180 kD and 160 kD. These proteins are detected only in haemolymph and ovarian extracts of vitellogenic females. The molecular weight of the native protein determined by size exclusion chromatography is approximately 400–420 kD.
A highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed to monitor the temporal changes in vitellogenin titre in haemolymph. Vitellogenin production starts on day 2 of the last larval stadium, reaching a maximum level by day 6 of the last larval stadium, and decreasing in the late pupal stage as vitellogenin was internalized into the oocytes. This is the first report of vitellogenin production occurring in the larval stage of a holometabolous insect. The fact that vitellogenin production and uptake occur during different stages of development in the gypsy moth, opens up some interesting questions concerning the underlying regulatory mechanisms controlling each process.     

Sperm utilization and post-copulatory female-guarding in the Colorado potato beetle, Leptinotarsa decemlineata

Crosses between white and black color morphs of the Colorado potato beetle, Leptinotarsa decemlineata Say, used as genetic markers, showed that male beetles ride and guard females to achieve the three copulations required to fill the spermatheca. Multiple matings are necessary for the females to realize their full reproductive potential. Sperm mixing occurs in the spermatheca providing partial sperm precedence. The data support the competitive mate searching theory.

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Résumé Différentes combinaisons d'accouplements entre des mutants noirs et blancs du doryphore de la pomme de terre, Leptinotarsa decemlineata, servirent à étudier le comportement de guarde de la femelle par le mâle après un premier accouplement. Les résultats indiquent que le sperme de deux mâles accouplant la même femelle se mélange dans la spermathèque assurant ainsi un certain niveau de précédence du sperme du dernier mâle.La masse de sperme dans la spermathèque était inférieure à sa capacité chez les femelles ayant été accouplées une ou deux fois. Le doryphore mâle aurait donc avantage à demeurer sur la femelle après un premier accouplement afin d'obtenir le minimum de trois accouplements requis pour remplir la spermathèque avant que les autres mâles puissent accoupler cette femelle. Les autres mâles ont avantage à essayer de dérober cette femelle pour remplir eux mêmes la spermathèque ou tout au moins profiter d'une certaine précédence du sperme du dernier mâle pour fertiliser une partie des oeufs.Sans accouplements multiples, la femelle ne possède pas suffisamment de sperme pour féconder tous ses oeufs. II semble aussi que les accouplements répétés puissent augmenter sa fécondité.Les résultats supportent une stratégie reproductive de compétition plutôt qu'une simple maximisation du nombre d'accouplements.

     

EFFECT OF TREE CANOPY REMOVAL BY GYPSY MOTH LARVAE ON THE MACROALGAE OF A RHODE ISLAND HEADWATER STREAM1

A spring-fed, headwater stream in central Rhode Island was examined during the period from June to October, 1979 to 1982. In the first two summers, a dense riparian canopy reduced the light penetration at the stream surface to a range of 5 to 18% of incident radiation. The lotic macroalgal community during this period was limited to 1 to 4 species covering < 1 to 35% of the stream bottom. However, in June and July, 1981, the surrounding leaf canopy was removed by a massive gypsy moth larval outbreak. Light penetration to the stream during this summer increased to 73% by early July, thereby resulting in a rise in water temperatures by 3.7°C. Even though there was a partial regrowth of leaves in late July and August of 1981, macroalgal cover values continued to rise to an early August peak of 80%. During the third summer, 88% of the macroalgal abundance could be attributed to illumination and water temperature. The filamentous diatom Funotia pectinalis ( O.F. Müll.) Rabh. was the predominant species in the midsummer of all four years, accounting for at least 60% of the total cover. In 1981. an important taxon was the desmid Hyalotheca dissiliens (S. Smith) Bréb., a species which was not seen in other years. A less severe gypsy moth defoliation occurred in 1982 but did not produce significant differences in light, temperature or macroalgal cover from 1979 and 1980. The results indicate that light and temperature can be limiting during the summer in spring-fed, headwater streams and that seed populations of some species are present in undetect-able levels during these periods of suboptimal growth conditions. In addition, it appears that stream macroalgal communities can be quite resilient, recovering rapidly following a major perturbation .     

Mitochondrial aldehyde dehydrogenase from higher plants

Aldehyde dehydrogenase has been purified to homogeneity from mitochondria of potato tubers and pea epicotyls. Although the enzyme had a high affinity for glycolaldehyde it also had a high affinity for a number of other aliphatic and arylaldehydes. It is proposed that the codification glycolaldehyde dehydrogenase (EC 1.2.1.22) should be abandoned in favour of mitochondrial aldehyde dehydrogenase (EC 1.2.1.3). The purified enzyme showed esterase activity and had properties similar to those reported for the mammalian mitochondrial aldehyde dehydrogenase. Although the natural substrate(s) for the enzyme is not known, the kinetic properties of the enzyme are consistent with it playing a role in the oxidation of acetaldehyde, glycolaldehyde and indoleacetaldehyde.     

Death of Cydia pomonella larvae and damage to apple fruit, after field application of codling moth granulosis virus

In field experiments, larvae of codling moth Cydia pomonella (L.) rarely acquired granulosis virus on hatching from the egg, but picked up most later, on the tree surface. Deposits of virus sprayed in 1.0% w/v skimmed milk did not affect neonate larval behaviour. Larvae died, usually in the first instar, after entering treated fruit, but they frequently entered via the calyx or near the base of the stalk or through cracks in the skin, where little feeding damage by first-and sometimes second-instar larvae was seen.

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Résumé En verger, la pulvérisation d'oeufs de carpocapse avec du virus de la granulose en suspension dans l'eau (additionnée de lait écrémé dilué à 1%) n'a pas modifié la survie des chenilles avant pénétration dans le fruit; par contre la pulvérisation des arbres a provoqué une forte mortalité. Bien que des chenilles consommant des poils et la surface des feuilles aient été observées avant leur pénétration dans le fruit, ce qui aurait pu provoquer leur contamination par le virus, il semble que la contamination létale provienne des fruits seuls.La présence de produit n'a modifié ni le comportement larvaire, ni le taux de pénétration dans les fruits; la mortalité y a lieu ensuite, généralement au premier stade. Dans 74 à 78% des cas, les chenilles ont pénétré dans le fruit par le calice ou près de la base du pédoncule — aucun dégât provenant de larves du premier stade n'y était visible, de même que dans le calice pour les larves du deuxième stade. Par contre, toute pénétration par la surface du fruit était repérable dès le premier stade. Il est possible que la répartition des lieux de pénétration dans le fruit influe sur la létalité due au virus et explique les variations d'efficacité observées en verger. Un système de classification des dégâts, provoqués lors de la pénétration dans le fruit, de chenilles du premier au troisième stade est proposé pour évaluer l'efficacité des essais en verger.