THE FINE STRUCTURE OF DIFFERENTIATING SPERMATOZOA AND SERTOLI CELLS IN THE GONAD OF THE POND SNAIL, LYMNAEA STAGNALIS

The fine structure has been examined, of spermatogonia, spermatocytes,early spermatids, late spermatids and early spermatozoa nestlingagainst Sertoli cells in the gonad of Lymnaea stagnalis. Changes in the Sertoli cells are linked with the phases of spermdifferentiation. Details on differentiation particularly ofthe head of the sperm, are presented. (Received 14 March 1981;     

Ultrastructural Aspects of the Glandular Cells from the Secretory Trichomes and from the Cell Suspension Cultures of Achillea millefolium L. ssp. millefolium

The ultrastructure of the glandular cells of the floret secretorytrichomes from Achillea millefolium L. ssp. millefolium (yarrow)was examined before and after anthesis and compared with theultrastructure of the cells from the cell suspension culturesobtained from the same plant. The profuse tubular structuresobserved in the plastids of the glandular cells of the trichomesduring the pre-secretory stage were much reduced in the secretorystage and showed an osmiophilic content. Some endoplasmic reticulumprofiles could be seen adjacent to the plastids. Later in thesecretory stage, the secretion appeared in the periplasmic spacebetween the cells of the upper tiers and in the sub-cuticularspace. Finally the secretion was released by rupture of thecuticle. At the lag phase, the cells from the cell suspensioncultures of yarrow were characterized by the presence or plastidswith tubular structures, similar to those observed in the plastidsof the trichomes in the pre-secretory stage. By the end of thelag phase accumulations of starch were observed inside the plastids.At the beginning of the exponential phase, the tubular structuresof the plastids started to show an osmiophilic content and theaccumulations of starch were still present. At the end of thisphase starch disappeared from the plastids and only osmiophilictubular structures were observed. Rough endoplasmic reticulumas well as smooth endoplasmic reticulum profiles were frequentlyin close association with plastids and mitochondria. At thestationary phase a very large vacuole filled the cells, andin the remaining cytoplasm some endoplasmic reticulum profilesand osmiophilic droplets were observed.Copyright 1994, 1999Academic Press Achillea millefolium L. spp. millefolium, yarrow, ultrastructure, trichomes, glandular cells, plant cell suspension cultures     

STUDIES ON CHLOROPHYLLASE OF CHLORELLA PROTOTHECOIDESI. ENZYMATIC PHYTYLATION OF METHYL CHLOROPHYLLIDE

1. The relation between chlorophyll content and the hydrolyticactivity of chlorophyllase in Chlorella protothecoides was examined.An increase in the activity was parallel to that in chlorophyllcontent during the development of green colouration, or greeningcourse, in the bleached cells. The activity sharply declinedand a parallel disappearance of chlorophyll was also found duringbleaching of the green cells.
2. A partially purified water-solublepreparation of chlorophyllasewas obtained by n-butanol treatmentand fractionation with coldacetone. It showed high activityand hydrolyzed 2 mg chlorophylla per hr per mg protein.
3. Forseparation and identification of the pigments concernedin thechlorophyllase reaction, a new solvent system of paperchromatographywas introduced.
4. When methyl chlorophyllide a and phytol wereincubated withthe enzyme, two products were formed. By comparisonwith theRf values of isolated pure substances, one was identifiedaschorophyll a and the other as chlorophyllide a. This enzymedid not catalyze the phytylation of free chlorophyllide a, butit had the ability to attach phytol to methyl chlorophyllidea. The final step in the biosynthesis of chlorophyll a is brieflydiscussed.

¹ Contribution No. 158 from the Department of Biology, Facultyof Science, Kyushu University. Supported in part by a grant-in-aidfor Fundamental Scientific Research from the Ministry of Education.     

苦皮藤素V对东方粘虫中肠细胞及其消化酶活性的影响

苦皮藤素V是从杀虫植物苦皮藤Celastrus angulatus Max.根皮中分离的一种对昆虫具有毒杀活性的新化合物。该文通过电镜观察和生化分析研究了其对东方粘虫Mythimnaseparata(walker)幼虫中肠组织及中肠主要消化酶活性的影响。电镜观察发现,中毒试虫的中肠细胞及其细胞器发生明显病变：柱状细胞顶膜微绒毛零乱、减少;线粒体肿胀,出现空白亮区,双层膜不完整;细胞质密度降低,细胞器排列紊乱;内质网池扩张,囊泡化,粗面内质网减少;杯状细胞杯腔变大,微绒毛减少。消化酶活性测定结果表明,中毒试虫中肠的蛋白酶、淀粉酶及脂肪酶的活性和正常虫相比,无显著变化。因此认为,苦皮藤素V主要作用于中肠细胞的质膜及其内膜系统。     

ALLOZYME COMPARISONS AMONG SPECIES OF THE BULINUS FORSKALII GROUP (GASTROPODA: PLANORBIDAE) IN CAMEROON

Allozyme comparisons were made for adult Bulinus forskalii,B. camerunensis, and B. senegalensis from populations in Cameroonusing starch gel electrophoresis. Fifteen loci were examinedbut no differences were found between B. forskalii and B. camerunensis,bringing into question the validity of B. camerunensis as aseparate species. Bulinus senegalensis differed from B. forskaliiand B. camerunensis in allozymes for acid phosphatase, alpha-glycero-phosphatedehydxogenase, hydroxybutyrate dehy-drogenase, and phosphoglucomutase.No differences were found, as reported elsewhere, between B.forskalii and B. senegalensis for aspartate amino-transferase,isocitrate debydrogenase, phosphogluco-isomerase, and xanthineoxidise. No polymorphism was seen in B. camerunensis or variousB. senegalensis populations examined. Three alleles for isocitratedehydrogenase were observed for B. forskalii. Mixed populationsof B. forskalii and B. senegalensis were found at four sitesbut no evidence of hybridization between these species was found. (Received 9 May 1988; accepted 30 August 1988)     

SPECIFIC RNA FROM PHOTOPERIODICALLY INDUCED COTYLEDONS OF PHARBITIS NIL

The nucleotide ratio of several RNA species from cotyledonsof Pharbitis nil subjected to a single 16 hr night with or withouta 15 min light-break, or to continuous light was investigated.RNA species examined were RNAs from nuclear, mitochondrial,microsomal, and supernatant fractions separated by differentialcentrifugation, and s-, r-, and m-RNAs fractionated by methylatedalbumin column chromatography. Of the RNAs examined, m-RNA alone was found to change its nucleotideratio with photoperiod applied. Thus as compared with m-RNAfrom non-induced cotyledons (exposed to continuous light oran interrupted night), m-RNA from cotyledons induced by an uninterruptednight contained significantly reduced guanylic and cytidylicacids on molar ratio basis. A working hypothesis was proposed that floral stimulus productionin cotyledons may be directed by gene DNA derepressed photoperiodically. (Received October 18, 1966; )     

An Electron Microscope Investigation into the Structure of the Floridean Pit

Ultra-thin sections of the red algae Rhodymenia palmata andLaurencia caespitosa were examined in the electron microscopein order to determine the fine structure of the Floridean pits.The pits found in the two species examined are shown to be quitedifferent from each other, in Rhodymenia the pit is open sothat the cytoplasm is continuous through the cell wall, whilstin Laurencia the pit is completely closed by a lamella of thecell wall. These two types are shown to be equivalent to thosedescribed by Jungers (1933) on the basis of light microscopeobservations.     

DIFFERENTIATION OF SERTOLI CELLS AND POST--REPRODUCTIVE EPITHELIAL CELLS IN THE HERMAPHRODITE GLAND OF ARION ATER(L.) (MOLLUSCA, PULMONATA)

Differentiation of Sertoli cells and post-reproductive epithelialcells during development of the hermaphrodite gland has beendescribed in Arion ater (L.) at the fine structure level. Twodifferent stages of Sertoli cells, active and atrophic, arerecognized. The cytoplasm during the active stage shows highmetabolic activity by many lysosome-like inclusions and otherorganelles. The possible role of steroidogenesis of these cellsis discussed in relation to their fine structure. The post-reproductive epithelial cells appear in the gland approximatelyafter the sheddingof both spermatozoa and oocytes. These cellsmay fill the lumen of the gland and appear syncytial. ^*Present address: Department of Biology, School of Sciences,Mashad University, Mashad, Iran (Received 12 January 1979;     

Visualization of nerve fibers and taste buds of the tropical catfish, Corydoras aeneus, following treatment by trypsin

Taste buds were removed from the barbels of the tropical catfishCorydoras aeneus following fixation with a 10% solution of formalinand subsequent treatment with a 0.25% solution of trypsin. Individualepithelial cells were dissociated as free cells. The surfaceof the dermis, including dermal papillae, appeared smooth andwavy, but some undissociated epithelial cells remained on thesmooth surface of the dermis. The surface of the dermis withoutfixation appeared fibrous. A few fine bundles of nerve fibersprotruded from the perimeter on the top of dermal papillae.These observations show that fine bundles of nerve fibers penetratethrough the remaining basement membrane at several points alongthe perimeter on the top of the dermal papillae.     

Above- and Below-ground Production of Young Scots Pine (Pinus sylvestris L.) Trees after Three Years of Growth in the Field under Elevated CO2

Scots pine (Pinus sylvestris L.) seedlings were grown for 3years in the ground in open top chambers and exposed to twoconcentrations of atmospheric CO₂(ambient or ambient + 400 µmol mol^-1) without addition of nutrients and water. Biomassproduction (above-ground and below-ground) and allocation, aswell as canopy structure and tissue nitrogen concentrationsand contents, were examined by destructive harvest after 3 years.Elevated CO₂increased total biomass production by 55%, reducedneedle area and needle mass as indicated, respectively, by lowerleaf area ratio and leaf mass ratio. A relatively smaller totalneedle area was produced in relation to fine roots under elevatedCO₂. The proportion of dry matter in roots was increased byelevated CO₂, as indicated by increased root-to-shoot ratioand root mass ratio. Within the root system, there was a significantshift in the allocation towards fine roots. Root litter constituteda much higher fraction of fine roots in trees grown in the elevatedCO₂than in those grown in ambient CO₂. Growth at elevated CO₂causeda significant decline in nitrogen concentration only in theneedles, while nitrogen content significantly increased in branchesand fine roots (with diameter less than 1 mm). There were nochanges in crown structure (branch number and needle area distribution).Based upon measurements of growth made throughout the 3 years,the greatest increase in biomass under elevated CO₂took placemainly at the beginning of the experiment, when trees grownin elevated CO₂had higher relative growth rates than those grownunder ambient CO₂; these differences disappeared with time.Symptoms of acclimation of trees to growth in the elevated CO₂treatmentwere observed and are discussed. Copyright 2000 Annals of BotanyCompany Elevated CO₂, Pinus sylvestris, biomass production, allocation, fine roots, root litter, crown structure, nitrogen, C/N ratio     

PHYSIOLOGICAL.STUDIES OF A CHIMAERA FORMED BY GRAFTING NIGHTSHADE UPON TOMATO I. RESPIRATORY ACTIVITY OF LEAVES

1. Leaves of a chimaera which are composed of the tissues fromLycopersicum esculentum var. kinnari (L. e.) covered by twolayers of cells from Solatium villosum (S. v.) were comparedwith those of control plants, L. e. and S. v. with respect tothe respiratory activity. 2. As for the respiratory activity of leaf strips, the chimaerawas more active than either L. e.. or S. v. 3. To respiratory inhibitors examined leaf strips of the chimaeraresponded rather similarly to those of S. v. but differentlyfrom those of L. e. 4. Of various substrates tested, glycolic, ascorbic and chlorogenicacids were actively oxidized by cell-free extracts of leavesfrom every plant. The first two were especially good substratesfor S. v.: L. e. attacked most the third, and the chimaera oxidizedjust as well the first two as S. v. did and the third as L.e. did. 5. Ascorbic acid and chlorogenic acid contents of the leaf extractswere not correlated with the activities of the extracts of oxidizingthese substances. 6. Physiological interactions were suggested between the twotissues of different origins in the chimaera. (Received December 2, 1963; )     

距树干不同距离处华北落叶松人工林细根生物量分布特征及季节变化

 该文研究了华北落叶松(Larix principis-rupprechtii)人工林细根生物量水平分布和季节变化特征。采用钻土芯法(土钻内径7.0 cm), 在距树干20、50和100 cm处设取样点, 每个样点处分3层(0~10、11~20和21~30 cm)钻取土芯, 取样时间为5、7、9和10月。华北落叶松人工林细根(≤2 mm)生物量全年平均值为224.89 g•m^–2, 在水平分布上表现为100 cm处细根生物量最大(244.20 g•m^–2), 其次为20 cm处(221.03 g•m^–2), 50 cm处最少(209.45 g•m^–2)。在0~30 cm土层, 总细根(包括活跟和死根)生物量季节变化范围在169.67~263.09 g•m^–2之间, 9月细根生物量最大, 5月细根生物量最少。0~10 cm土层细根生物量季节变化差异显著(p＜0.05), 11~20和21~30 cm差异不显著(p＞0.05)。距树干100和20 cm处(0~10 cm土层), 细根生物量的季节变化差异明显(p＜0.05), 9月总细根生物量最大(172.82和185.68 g•m^–2), 5月总细根生物量最少(69.28和73.47 g•m^–2); 50 cm处季节变化差异不明显(p＞0.05)。细根生物量分布和季节变化不仅受土壤垂直格局影响同时也与距树干不同水平距离有很大的关系。     

THE FINE STRUCTURE AND FUNCTION OF THE SALIVARY GLANDS OF NUCELLA LAPILLUS (GASTROPODA: MURICIDAE)

The fine structure of the tubular and acinous salivary glandsof Nucella lapillus (L.) has been studied and some histochemicaland enzyme tests have been carried out. The clusters of subepithelialcells of the tubular glands secrete a glycoprotein composedof chains of tubular macromolecules resembling secretions knownto have adhesive properties which may assist in boring. Thesecretion is rich in disulphide groups, as are many toxins,and is believed to be responsible for the recently demonstratedpharmacological activity of the glands. It is proposed thatflaccid paralysis is induced in prey by envenomation with thissecretion during rasping, after soft parts have been exposedby an ‘anti-predator’ reaction to secretion fromthe hypobranchial gland of Nucella. The secretory vesicles ofboth types of gland cells in the acinous glands have heterogeneouscontents indicating that their secretions are complex. The majorcomponent in those of the mucous cells is an acid mucopolysaccharidetypical of a lubricant or releasing agent. The ciliated basalcells resemble typical enzyme-secreting cells and the majorconstituent of their secretion is a finely granular glycoprotein. (Received 8 January 1990; accepted 5 June 1990)     

SYNCHRONIZATION OF BACTERIAL CULTURE BY PREINCUBATION OF CELLS AT HIGH CELL DENSITY

The growth of Escherichia coli strain B in a liquid medium wasfound to cease at a cell density of 5x10⁹ cells per ml. (Thiscritical concentration is designated as the maximum or M-concentration.)Even cells harvested from the logarithmic growth phase couldnot divide at this or higher cell densities. Investigationson the metabolic activities of such cultures, however, showedthat the synthesis of cellular protein and nucleic acid wastaking place under such circumstances, showing that only someprocess (or processes) particularly related to cell divisionwas suppressed at the critical cell concentration in question. This finding led us to devise a new method of synchronizationof E. coli: cells harvested from a logarithmic phase were preincubatedat the critical concentration of 5x10⁹ cells per ml for 45 minutes,and then diluted 100 times with fresh medium. This led to synchronizationof cell division, as shown by a stepwise multiplication in cellnumber. (Received June 20, 1961; )     

COMPARATIVE STUDIES OF PHOTOCHEMICAL OXIDATION-REDUCTION REACTIONS IN LAMELLAR FRAGMENTS OF VARIOUS ALGAE AND SPINACH

The activity of various electron carriers, including DPIP, spinachplastocyanin, mammalian cytochrome c, and Anabaena cytochrome553, as donor in the reaction induced by the photochemical systemI was examined with lamellar fragments of various algae andspinach. Reduced DPIP was an effective electron donor irrespective ofthe organisms, when it was supplied at a high concentration(10^–3 M). Spinach plastocyanin was effective in the reactionswith the lamellae of green algae, Euglena, diatom Phaeodactyrumand red algae Porphyra yezoensis and Porphyra sp. Yamamoto II,whereas it was inactive in the lamellae of blue-green algae.Horse-heart cytochrome c and Anabaena cytochrome 553 were activein the reaction with the lamellae of bluegreen algae. The formercytochrome was also active in the reactions in Porphyridiumand Cyanidium. The cytochromes were less active in the reactionsin which spinach plastocyanin acted as effective electron donor. The data were interpreted as that the photochemical system Iin bluegreen algae differs from that of other photosyntheticorganisms with respect to the properties of the site of theelectron-input. ¹ Present address: Nomura Research Institute for Technologyand Economics, Kamakura, Kanagawa. ² Present address: Ocean Research Institute, University of Tokyo,Nakano, Tokyo.     

"GIGANTISM" OF CHLORELLA VULGARIS I. MECHANISM OF INDUCTION OF CIGANTISM

1. Based on the microscopic observations, two stages, "giant cellstage" and the subsequent "palmelloid body stage", were distinguishedin the process of formation of giant Chlorella induced by theaddition of sugars. The "giant cell" is much larger in sizethan the control cell, but the other morphological featuresare the same as those of the latter. The "palmelloid body" isa form composed of many conjoined autospores.
2. When a highconcentration of glucose was maintained in the medium,gigantismwas also maintained. Under this condition, the algashows acyclic transformation between "giant cell" and "palmelloidbody"without returning to the small single cells.
3. Large amountsof carbohydrate composed of hexose were foundto be accumulatedin the giant algal cells, and it was inferredthat this carbohydrateaccumulation causes greater enlargementof cell volume as comparedwith control cells.
4. Uronic acids, which were found to be absentin the control cells,were formed and lost in the cells culturedin the glucose mediumin parallel with the appearance and disappearanceof gigantism.
5. Pectic substances, from which uronic acids areconsidered tobe derived during the extraction procedure, werefound to bepresent only in giant Chlorella.
6. The conjoinedautospores in giant Chlorella (at the palmelloidbody stage)were separated to some extent by the addition ofEDTA, and theresulting cells were similar to control Chlorellacells.
7. Basedon these results it was inferred that inductive formationofthe pectic substances is causally related with the appearanceof "palmelloid body".

¹ Present address: Department of Chemistry, College of GeneralEducation, Osaka University, Toyonaka, Osaka.     

AN ULTRASTRUCTURAL STUDY OF THE GLAND OF LEIBLEIN OF MURICID AND NASSARIID NEOGASTROPODS IN RELATION TO FUNCTION, WITH A DISCUSSION ON ITS HOMOLOGIES IN OTHER CAENOGASTROPODS

The gland of Leiblein of the muricid Nucella lapillus and thenassariid Hinia reticulata has been examined by scanning andtransmission electron microscopy. The origin and functionalsignificance of its complex organization and its relationshipwith the rest of the mid-oesophagus in Nucella are discussed.It is absorptive as well as secretory, and a mechanism is proposedby which solute-rich fluids may enter the gland. Its epitheliumis composed of occasional mucous cells and two major cell types:ciliated cells engaged in protein metabolism and unciliatedcells responsible for uptake and storage of lipids and carbohydrates,both of which show evidence of pinocytotic uptake of solutesand intracellular digestion in lysosomes. Some enzyme activitypersists in the residual bodies they shed by apocrine secretion,but they remain intact in a mucous string until they reach thestomach. Preliminary ultrastructural examination indicates thatthe gland absorbs cadmium not only from the blood but also directlyfrom its lumen and that it may have the capacity to sequestera wide range of toxins. The same types of cell occur in Hiniain which their cyclical activity has been correlated with feeding.Similar cells have been identified in the oesophageal glandsof other prosobranchs. The foregut glands of carnivorous caenogastropodsare compared with the gland of Leiblein. There is an inversecorrelation between the role of the mid-oesophagus in digestionand absorption and the complexity of the stomach. (Received 16 August 2004; accepted 24 January 2005)     

Characterization of Plant Epidermal Lens Effects by a Surface Replica Technique

Optical replicas of leaf surfaces were made for characterizingthe lens properties of individual epidermal cells. Using a dentallatex, moulds were made of leaf surfaces and subsequently usedto produce agarose replicas. The replicas focused light in amanner similar to intact epidermal cells and it was possibleto measure both focal lengths and intensifications within leafreplicas of Thermopsis montana, Mahonia repens, and Smilacinastellata which had epidermal cells of different diameter. Focallengths ranged from 74—130 µm which indicated thatlight was concentrated within the underlying photosynthetictissues of these leaves. Focal intensifications were measuredsensiometrically and were 1.5 for T. montana and 2-6 for theother species. These values compare favourably with calculatedfocal lengths and measurements taken from isolated epidermallayers. The results indicate that the epidermis can concentratelight within the leaf to amounts well in excess of ambient light.Furthermore, the replicas faithfully reproduced fine anatomicaldetails from a wide variety of leaves and they provide a non-destructiveway to reproduce surface characteristics for anatomical andphysiological studies.     

The Structure of Mature Rye Endosperm

Endosperm tissue of mature kernels of rye (Secale cereale L.)cv. Dominant was examined by light and transmission electronmicroscopy. It was found that storage protein in sub-aleuronecells occupies up to 35 per cent of the cell volume and formsa continuous matrix in which starch grains and cytoplasmic remnantsare embedded. In the prismatic endosperm, the storage proteinis present as a fine network interspersed between the numeroustype A and B starch grains. Protein bodies are not found inthe prismatic endosperm; only a few, less than 1 µm indiameter, are observed in pockets of disorganized cytoplasmin the sub-aleurone tissue. Thick cell walls and intercellularmaterial may contribute to the high pentosan content of ryeendosperm. Secale cereale L., rye, endosperm, protein matrix, ultrastructure     

A COMPARATIVE STUDY OF CHROMOSOMES IN FOUR SPECIES OF THEODOXUS (GASTROPODA: NERITIDAE)

Karyological analysis was performed on Theodoxus baeticus (Lamarck,1822), T. valentina (Graells, 1846), T. velascoi (Graells, 1846)and T. fluviatilis (Linne, 1758), collected from freshwaterbodies of eastern Spain. Cells possessing diploid chromosomenumbers of 2n 5 25 were most common in the tissues of males,whilst 2n 5 26 was most prevalent in females. The sixth pairof chromosomes had only one homologue in mitotic cells of males,indicating the XO/XX sex-determining mechanism. The absolutelength of chromosomes ranged from 1.65 to 6.33 mm. The relativesizes of chromosomes varied from 3.92 to 13.33% of the totalhaploid set length. In all species chromosome pairs no. 1, 4,7-13 were composed of metacentric, pairs no. 5 and 6of submetacentricchromosomes. Pairs no. 2 and 3 were submetacentric, subtelocentricor submeta- subtelocentric according to the species studied.There were significant differences (P, 0.05) among the relativelength, as well as centromeric indices of different chromosomepairs across the species. Karyological differences were greatestbetween T. fluviatilis and three other Theodoxus species. (Received 6 March 2000; accepted 8 May 2000)