Phytochrome B Is Not Detectable in the hy3 Mutant of Arabidopsis, Which Is Deficient in Responding to End-of-Day Far-Red Light Treatments

Immunoblot analysis using a monoclonal antibody specific tophytochrome B (phyB) suggest that this protein is undetectablein the hy3 mutant of Arabidopsis. Photophysiological experimentsrevealed that hy3 mutants do not display end-of-day far-redgrowth responses. Thus, this particular phytochrome responseseems to be mediated in large part by phyB. (Received September 20, 1991; Accepted September 26, 1991)     

Ultrastructural Studies on Development in Mature Seeds; Embryos of Euonymus europaea L. Dormant or Cultured at +25 ?C

Ultrastructural observations on naturally dispersed and dehydratedseeds of Euonymus europaea L. were performed. It was shown thatduring the culture of isolated embryos at 25 ?C for 7 d thematuration of protein bodies continued; the initial juvenileform (single protein body per cell with dispersed contents)was developing into a more advanced form (single body splitto form several sub-units with condensed contents). Parallelbiochemical determinations pointed to an increase in insolubleprotein levels. There were no changes in other storage organelles,lipid bodies, either in the fine structure or in the level oftriacylglycerols. Deterioration of cellular membranes (in mitochondria,proplastids and protein bodies) similar to that described forageing was observed in cells at the periphery of the embryonicaxis. It is concluded that during the culture of E. europaeaembryos the maturation programme of development in protein bodiesis being continued. Key words: Embryonic axis, storage protein and lipid body, maturation     

AMNICOLA CALLOSA PAULUCCI, 1881, REDESCRIBED AND ASSIGNED TO ORIENTALINA RADOMAN, 1978, A GENUS NEW FOR THE ITALIAN FAUNA (GASTROPODA, PROSOBRANCHIA, HYDROBIIDAE)

Anatomical study of topotypes of Amnicola callosa Paulucci,1881 from Abruzzo (Central Italy) produced evidence that thespecies belongs to Orientalina Radoman, 1978, a genus previouslyonly known from the Balkan Peninsula. (Received 26 June 1991; accepted 20 August 1991)     

THE FINE STRUCTURE AND FUNCTION OF THE ANAL GLAND OF THE MURICID NUCELLA LAPILLUS (NEOGASTROPODA) (AND A COMPARISON WITH THAT OF THE TROCHID GIBBULA CINERARIA)

The fine structure of the anal gland of Nucella lapillus isdescribed, and compared with that of Gibbula cineraria. It isconcluded that the glands are not analogous in function andare unlikely to be homologous. The anal gland of Gibbula isa small rectal diverticu-lum near the anal papilla, lined bymucus-secreting cells. The evidence suggests that the largeanal gland of Nucella extracts macromolecules and cations fromthe blood, metabolizes or sequesters them in lyso-somes andultimately expels the resultant residual bodies by apocrinesecretion. It has large reserves of lipid and glycogen, andthe abundance of melanin indicates that it is a major site oftyrosine degradation. The invariable presence of bacteria inthe lumen, and of pits in the epithelium to house them, impliesa symbiotic relationship in which the bacteria metabolize anddegrade the cell debris, some of which is resorbed. The lossof the anal gland in bucci-nids and nassariids is accompaniedby great reduction in size and importance of the rectal sinus,and increase in complexity of the kidney. (Received 31 December 1991; accepted 23 January 1992)     

EXTRA-CAPSULAR YOLK BODIES IN THE EGG MASSES OF SOME TROPICAL OPISTHOBRANCHIA

Seventeen species of opisthobranch molluscs from Enewetak, MarshallIslands were found to deposit bodies of yolk within their gelatinousegg masses external to the capsules which surround the ova.These observations bring the total number of opisthobranch speciesin which extra-capsular yolk is known to 26. This study reviewsprevious reports of extra-capsular yolk and contributes newdata on the egg masses and developmental characteristics ofspecies in which it is known to occur. The various types ofyolk body found are detailed and categorized, and original observationson the deposition of yolk bodies and their fate during, andsubsequent to embryonic development presented. To date, extra-capsular yolk bodies have been found only withinthe egg masses of species of Chromodoris and Cadlinella (Nudibranchia:Chromodorididae), Elysia (Sacoglossa: Elysiidae), and Bosellia(Sacoglossa: Polybranchiidae). Three types of yolk body arerecognized: (1) cap-like yolk bodies associated with individualcapsules, (2) discrete yolk bodies strewn throughout the eggmass, and (3) continuous or discontinuous yolk strings. Direct consumption of yolk bodies by newlyhatched veligers ofChromodoris albopunctatus (Garrett) is described. In most otherspecies, the yolk disappears during the course of developmentand is probably of nutritive value to the embryos. The availabledata suggest that chromodorid species which utilize extra-capsularyolk during the course of development produce relatively largerlarvae than species with comparably sized ova that do not. (Received 25 November 1982;     

Pigment Bodies in Fruits of Crimson and High Pigment Lines of Tomatoes

Pigment bodies in fruits of crimson (og^c), high pigment (hp),and crimson-high pigment (og^c hp) lines of tomatoes were observedby electron and light microscopy and compared with those ofnormal red lines and a yellow cultivar. During chloroplast-chromoplasttransformation, two main structurally distinct bodies are produced,their total and relative amounts apparently accounting for theentire range of colours (from very deep red to yellow) characterizingthe mature fruits of these different colour lines. The longnarrow crystalloids, believed to be lycopene, form in associationwith an extended thylakoid system; in senescing (over-ripe)fruit many of these are reduced to shorter irregular forms.The rounded globules are believed to be beta-carotene dissolvedin lipid material derived from membrane lysis. Analytical resultscorroborate microscopic observations that the effect of theog^c gene, as compared with the r⁺ gene for normal red colour,is to increase the lycopene content and lower the beta-carotenecontent. The effect of the hp gene is to increase the levelsof both pigments. The results support the view that the genescontrol the development of fruit pigments which affect chromoplastultrastructure. Lycopersicon esculentum Mill, tomato, fruit, pigment bodies, beta-carotene, lycopene     

Structural Characteristics and Fatty Acid Composition of Psophocarpus tetragonolobus Seeds

Winged bean (Psophocarpus tetragonolbtis) seed coat in the hilarregion consists of a double layer of sclereids, tracheid barand spongy parenchyma cells. This is contrasted to the seedcoat structure on either side of the hilar region, which hasa single layer of sclereids, columnar cells and crushed parenchymacells. Cotyledonary cells are large (50 to 100µm in diameter)and have cell walls 2·4 to 4·7 µm thickwith pit-pair structures. Protein bodies and lipid bodies arethe main structural components of the cytoplasm while only asmall number of starch granules are present in each cell. Themajor portion of the lipid can be removed by non-polar solventsand contains oleic and linoleic acids as the predominant unsaturatedfatty acids. High levels of behenic acid were present in both‘free’ and ‘bound’ lipids. Psophocarpus tetragonolobus (L.) DC., winged bean, seed structure, seed coat, protein bodies, lipid bodies, fatty acids, oleic acid, linoleic acid     

Ultrastructural Changes in Cotyledons of Pineapple Guava (Myrtaceae) During Somatic Embryogenesis

Ultrastructural studies (SEM and TEM) were performed on cotyledonsof pineapple guava ( Feijoa sellowiana Berg, Myrtaceae) inducedto form embryos on medium containing 1.0 mg l^-1(4.5µM2,4-D) and 0.3M sucrose. At the time of culture, the cells werefilled with protein and lipid bodies. Microbodies and poorlydifferentiated organelles could also be seen. In contrast togerminating cotyledons, where lipid and protein reserves werequickly metabolized, cells of the embryogenically induced cotyledonsshowed evidence of reserve consumption only after 5 d of culture.Subepidermal cells of the upper cotyledonary surface underwentseveral divisions giving rise to a meristematic layer of severalcells thickness from which somatic embryos developed. Embryoscould also be formed directly by successive divisions of epidermalcells. Cells involved in somatic embryo formation containeda large nucleus with a conspicuous nucleolus and dense cytoplasmwhere numerous ribosomes, mitochondria, plastids with starchand short profiles of rough endoplasmic reticulum were present.Plasmodesmata were present both in cell walls of the meristematiccells and in few celled embryos whereas in degenerating embryosor in more advanced stages of somatic embryo development noplasmodesmata could be found. Although oil bodies were not observedin the meristematic cells they were identified in very youngembryos, being the first reserve compounds to appear. Cellsnot involved in somatic embryo differentiation were characterizedby the presence of several microbodies containing a crystalloidinclusion and elongated mitochondria. Feijoa sellowiana ; pineapple guava; somatic embryogenesis; ultrastructural studies     

Lipids in Cryptomonas CR-1. II. Biosynthesis of Betaine Lipids and Galactolipids

The biosynthesis of lipids in Cryptomonas strain CR-1 was studiedusing radioactive tracers. For studies of general aspects ofthe biosynthesis of lipids, the cells were labelled with [¹⁴C]NaHCO₃or with [l,3-¹⁴]glycerol. In both cases, monogalactosyl diacylglycerol(MGDG) was the most heavily labelled lipid. Phosphatidylcholineand the alanine lipid DGTA were not labelled to specific activitiescomparable to those of MGDG and DGDG. It is improbable thatthe so-called "eukaryotic pathway", which has been suggestedas the pathway for the synthesis of " eukaryotic" molecularspecies of MGDG from PC in higher plants, is operative in Cryptomonascells which contain typical "eukaryotic" MGDG. The homoserinelipid DGTS was labelled to a significant level only in its polargroup. The C-3 and C-4 atoms of methionine, as well as the methylcarbon of methionine, were incorporated into both DGTS and DGTA,whereas the C-l carbon of methionine was incorporated uniquelyinto DGTS. Results of pulse-chase experiments with [3,4-¹⁴C]methionineand [methyl.-^l4C]methionine suggest the conversion of DGTS toDGTA. (Received April 22, 1991; Accepted June 12, 1991)     

Distribution of Diacylglycerylhydroxymethyltrimethyl-{beta}-alanine (DGTA) and Phosphatidylcholine in Brown Algae

Lipids were analyzed in thirteen species of brown algae collectedat the seashore near Tokyo, Japan. Diacylglycerylhydroxymethyltrimethyl-ß-alanine(DGTA), a recently identified betaine lipid, was found as amajor lipid component in eight species of brown algae examined,namely, Ishige okamurai, Dictyota dichotoma, Pachydictyon coriaceum,Padina arborescens, Hizikia fusiformis, Sargassum horneri, S.ringgoldianum and S. thunbergii. However, phosphatidylcholine(PC) was not detected in any of these algae except I. okamurai.By contrast, PC was found as a major lipid component in fiveother species, namely, Colpomenia sinuosa, Endarachne binghamiae,Scytosiphon lomentarius, Eisenia bicyclis, Undaria pinnatifida.These algae in turn did not contain detectable amounts of DGTA.The fatty acid composition of four selected species, S. lomentarius,U. pinnatifida, D. dichotoma and H. fusiformis, was also studied.The fatty acid components of DGTA in D. dichotoma and H. fusiformiswere similar to those of PC in U. pinnatifida, the major componentsbeing 16:0, 18:2 and 20:46 (also 16:1 in D. dichotoma). (Received December 14, 1990; Accepted April 10, 1991)     

Isolation and Biochemical Characteristics of Protoplasts of Jojoba (Simmondsia chinensis (Link) Schneider)

Two types of protoplasts were isolated from leaves of shootsor callus of subcultures of jojoba (Simmondsia chinensis (Link)Schneider). Protoplasts from leaves were rich in chloro-plastsand were about half the volume of protoplasts from callus. Theviability of preparations as determined by the Evans blue techniquewas 80%. From cell cycle analysis by flow cytometry of nuclei,leaf protoplasts were uniformly in a non-proliferating phase(G0-G1), while callus protoplasts presented many phases of thecell cycle. Protoplasts from calli had only half the Chi ofthose from leaves. Yet Chi a/b ratio, as well as protein andtotal lipid content per cell, were similar in both types ofprotoplasts. A major drop in polar lipids, chiefly in mono-and digalactosyldiacylglycerol, and a parallel increase in neutrallipids occurred during protoplast isolation. The 18:2/18:3 ratiodecreased in neutral lipids concomitant with an increase intriglycerides rich in linolenic acid. Our results suggest atriggering of lipolytic acylhydrolases during the protoplastisolation, as reported for other species. Plasmolysis of thecells with high osmolarity medium and long incubation timeswere required to get a good yield of jojoba protoplasts. Inthe course of this procedure water-deficit stress takes place.A parallel with lipid changes occurring under this type of stressis discussed. (Received April 22, 1991; Accepted July 3, 1991)     

Fine Structure of Peanut Root Nodules Induced by Nod+Fix- Strains of Bradyrhizobium with Special Reference to Lipid Bodies

Ineffective nodules of peanut induced by two nod⁺fix^–strains of Bradyrhizobium sp. were compared with the ones inducedby nod⁺fix⁺ strain NC92. One of the fix^– strains, 639is a transconjugate Tn5 mutant of NC92, while the other, 7091,is an isolate from ICRISAT soil. Both induce small nodules lackingleghemoglobin and nitrogen-fixing activity. Ultrastructuralobservations revealed that the nodules of 639, have enlargedperibacteroid space and lack persistence of nodule function.The 7091-induced nodules showed impediment in bacteroid releaseand differentiation. In both the ineffective nodules large amountsof lipid bodies were found to accumulate several times in excess,compared to the effective NC92 nodules. The large lipid accumulationin absence of nitrogen fixation supports the hypothesis thatin peanut nodules lipid bodies are utilized as a supplementarysource of carbon and energy for nitrogen fixation. Peanut, lipid bodies, nitrogen fixation, nod⁺fix^– Bradyrhizobium, ultrastructure     

A NEW, STYGOBIONT, VALVATIFORM, HYDROBIID GASTROPOD FROM AUSTRIA (CAENOGASTROPODA: HYDROBIIDAE)

A new, stygobiont species of Hauffenia (Pollonera, 1898) fromnortheastern Austria is described. It has been previously misidentifiedas H. kerschneri (Zimmermann, 1930), but detailed anatomicalinvestigations indicate that it is a separate species. Distinguishingcharacters are the very reduced peg of the operculum, a hypobranchialgland, an anterior lobe of the digestive gland in 25% of thespecimens, and an upright penial stylet. Some animals were infestedwith sporocysts and rediae. (Received 13 September 1991; accepted 1 November 1991)     

Aldolase--An Important Enzyme in Controlling the Ribulose 1,5-bisphosphate Regeneration Rate in Photosynthesis

This study was done to explore an enzymatic mechanism for thephotosynthetic carbon reduction cycle whereby the rate of synthesisof ribulose 1,5-bisphosphate (RuBP) could be changed while thelevels of intermediates other than 3-phosphoglycerate and RuBPwere kept constant. Chloroplast aldolase was purified to homogeneityfrom spinach leaves. When the enzyme was assayed in the directionof fructose 1,6-bisphosphate synthesis in the presence of theconcentrations of the substrates reported in vivo, the activitywas severely inhibited by physiological concentrations of RuBP.The aldolase reaction proceeded with a sequential mechanism.The K_m for dihydroxyacetone phosphate and D-glyceraldehyde 3-phosphatewere 0.45 mM and 40 µM, respectively. The activity wascompetitively inhibited by RuBP with respect to dihydroxyacetonephosphate. The K^I was 0.78 mM. The maximum activity of aldolasein spinach leaves was calculated as 1,360µmol (mg Chl)^–1h^–1 An equation to express the reaction for the synthesisof fructose 1,6-bisphosphate by aldolase was constructed topredict the metabolic rate of this reaction in vivo. The calculationclearly showed that aldolase is an important enzyme in controllingthe rate of RuBP regeneration. (Received March 25, 1991; Accepted August 12, 1991)     

Properties of a Large Complex with NADH Dehydrogenase Activity from Barley Thylakoids

When assays for NAD(P)H-ferricyanide oxidoreductases were performed,activities specific for NADH (0.23 unit (mg protein)^–1)and NADPH (0.68 unit (mg protein)^–1) were detected inchloroplasts isolated from leaves of barley (Hordeum vulgareL.). Activities of chloroplast NADH- and NADPH-ferricyanideoxidoreductase were 5-fold and 25-fold higher, respectively,than the maximum activity that could be attributed to mitochondrialcontamination. Moreover, most of the chloroplast NADH-ferricyanideoxidoreductase (60 to 80%) was solubilized by deoxycholate (DOC)from thylakoids as a single, high-molecular-mass complex thatwas distinguishable from the mitochondrial complex by its lowerelectrophoretic mobility in 3% polyacrylamide, as revealed byreduction of nitro blue tetrazolium (NBT) in the presence ofNADH or NADPH on gels after electrophoresis. The stroma yieldeda single band of a dehydrogenase (66 kDa) that used NADH asits electron donor. Several NADPH-dependent activities weredetected after electrophoresis of the stromal fraction. Moreover,chloroplast-specific activities could be distinguished frommitochondrial activities on the basis of the specificity ofthe donor and the acceptor of electrons, the dependence of theactivities on pH, and the sensitivity to various inhibitors.K_m values for NADH (26 µM) and NADPH (75 µM) werein the same range as those of mitochondrial activities. Mostof the NADPH-dependent activity probably corresponds to thechloroplast ferredoxin-NADP⁺ oxidoreductase. The possibilityis discussed that thylakoid NADH dehydrogenase(s) might be theproduct of chloroplast ndh genes and that this activity is involvedin chlororespiration. (Received April 25, 1994; Accepted December 5, 1994)     

Annual cycle of tintinnids in Admiralty Bay with an emphasis on seasonal variability in Cymatocylis affinis/convallaria lorica morphology

In water samples collected from the middle of Admiralty Bay(King George Island 62°08'S 58°25'W) between February1990 and January 1991, 17 Tintinnina species were noted. TotalTintinnina numbers in summer were very high (up to 5000 cellsm^–3), but species diversity was low, consisting mainlyof Cymatocylis affnislconvallaria, forma convallaria. Duringaustral winter, cell numbers were very low, but species compositionwas diverse. Cymatocylis affnislconvallaria, forma affinis,and C.affmis/convallaria, forma convallaria, the polymorphicforms of one species C.affinis/convallaria, appear to be interchangeableduring the year. The transition from one form to the other occursin the spring and autumn. The typical Tintinnina polymorphismcan be attributed to the prevailing environmental conditions.     

Effects of an Epidermal Growth Factor on the Growth of Zea Coleoptiles

Sections of the elongation zones of coleoptiles of Zea maysL. were incubated in buffers containing various concentrationsof human epidermal growth factor (EGF). The growth rate of sectionsincubated with 10, 1.0 or 0.1 fig liter¹ EGF was higher thanthat of control. EGF at 10 fig liter ^–1 increased thegrowth rate by 13% of the control rate. (Received September 25, 1990; Accepted June 19, 1991)     

An Analysis of Ultrastructural Changes during Oosphere-initial Development in Oogonia from Ca2+-sufficient and Ca2+-deficient cultures of Saprolegnia terrestris Cookson ex Seymour

The central vacuole system of oogonia of Saprolegnia terrestrisfrom Ca²⁺-sufficient cultures was fully enlarged prior to theformation of oosphere initials, which did not involve cleavagevesicles. In oogonia with fully-enlarged central vacuole systems,subsidiary vacuoles at the periphery of the system sometimescontained dense bodies, and dense-body profiles were sometimespresent within sections of the central vacuole system itself.As the central vacuole system enlarged, volume densities ofdense-body vesicles, peripheral vacuoles, lipid bodies and thecytoplasmic matrix decreased relative to total oogonial volume(peripheral protoplasm volume plus central vacuole volume),while the volume density of nuclei increased and that of mitochondriaremained constant. Relative to the peripheral protoplasm only,volume densities of dense-body vesicles, lipid bodies and mitochondriaincreased and volume densities of peripheral vacuoles and ofthe cytoplasmic matrix decreased, while the volume density ofnuclei increased during central vacuole enlargement but subsequentlydecreased during formation of oosphere initials. Under conditionsof Ca²⁺ deficiency, the volume densities of mitochondria andof the cytoplasmic matrix were significantly increased, whilethat of lipid bodies was significantly decreased, at early stagesof oogonial development; the volume densities of other organelleswere not significantly altered at any stage. Saprolegnia terrestris, oogonia, development, calcium, ultrastructure, stereology     

Lipid-ovary indices in food-limited Daphnia

The amount of lipid carried by a zooplankton individual is highlyvariable. Various lines of evidence have suggested that thenutritional condition of animals could be ascertained via thedetermination of lipid content. However, few studies have beenconducted where the lipid levels of animals of known conditionhave been measured. Here, we report on the lipid index and combinedlipid-ovary index values of Daphnia obrusa reared on foods madeup of varying quantity and quality of a unialgal (Scenedesmusacutus) diet. Lipid levels differed in different food types.However, in no cases were these indices positively correlatedto growth rate, such as would be necessary if one were to usethem to diagnose food limitation in the field. Thus, we urgecaution in the utilization of either the lipid or lipid-ovaryindex until more is known about the role of fatty acids in thediets of these animals.     

Age-related Changes in the Ultrastructure and Membrane Properties of Brassica napus L. Seeds

The ultrastructure was studied of imbibed non-aged winter rape(Brassica napus L.) seeds in comparison with that of artificiallyaged seeds in which viability was partially or completely impaired.In parallel, measurements were made of lipid-phosphorus content,the leakage of phosphate from the seeds and their vigour andgerminability. Decreases in lipid-phosphorus which accompaniedthe loss of viability corresponded to an increase in phosphateleakage, suggesting damage to cellular membranes. Three ultrastructuralsymptoms possibly related to age-induced membrane deteriorationwere observed: (i) the lowering of electron contrast in allcellular membranes excluding plasmalemma; (ii) coalescence ofsmall storage lipid bodies to larger units presumably as a resultof the degradation of enclosing half-unit membranes; and (iii)the appearance of protoplasmic inclusions inside the storageprotein bodies, possibly resulting from rupture of the enclosingunit membranes. It is suggested that the presence of enlarged fibrillar centresin nucleoli of low viability seeds observed here for the firsttime in aged seed material may be the morphological manifestationof age-induced damage to nucleic acids. Brassica napus L, seeds, accelerated ageing, ultrastructure, leakage