银杏营养贮藏蛋白质的亚细胞定位

在电子显微镜下,对银杏(Ginkgo biloba L.)枝条营养贮藏蛋白质的超微结构特征及在亚细胞水平的定位进行了系统研究.结果表明:银杏营养贮藏蛋白质主要存在于韧皮薄壁细胞的液泡内.银杏韧皮薄壁细胞内的营养贮藏蛋白质在细胞质内合成,由内质网膨大的槽库、质膜内折或高尔基体小泡发育形成贮藏蛋白质的液泡.液泡蛋白质主要以不定形块状、絮状或颗粒状形态存在.贮藏蛋白质在整个越冬期一直保持高含量,直到翌年春季萌芽时,贮藏蛋白质迅速转移再利用.随着新梢的生长,到了夏末秋初,又重新开始积累贮藏蛋白质.     

Seasonal changes in hepatocyte ultrastructure correlated with the cyclic synthesis of secretory proteins in the winter flounder (Pleuronectes americanus)

Liver tissue was sampled from flounder (Pleuronectes americanus) throughout the year with the intention of documenting changes in the ultrastructure coincident with the production and secretion of antifreeze proteins. In the winter, hepatocytes are dedicated to the production of these proteins and, in the female, also reproductive proteins. In both sexes, liver cells in the summer contain abundant lipid and glycogen stores. In the female, there is a conspicuous hepatocyte transformation from a fat-filled cell in the summer to one with well-developed rough endoplasmic reticulum in the winter. Large amounts of rough endoplasmic reticulum (11.2 mg/gm) were recovered after subcellular fractionation of female wintertime liver. The increased appearance of secretory organelles and the high number of nucleolar profiles observed in winter animals is consistent with the elevated demand for protein secretion and synthesis in both sexes. The fractional volumes occupied by lipid droplets and mitochondria were different when comparisons were made between sex and season. Females contained a greater volume of lipid than did males, and summer animals contained more lipid than those in winter.     

Studio Del Ritmo Vegetativo Di Olea Europaea L. in Messina Negli Anni 1952–54

Summary

The vegetative growth of the olive tree in Messina is described. The growth period begins at the end of March and prosecutes without interruption as late as November. The secondary wood tissue producted during summer time (from Yuly to August) is abnormal being of a parenchimatoide type. During winter there is a well defined rest period.

The cambium produces secondary wood and bark with an alternate rythm, so that in spring the wood production prevails on the liber and in autumn the opposite condition is realized.

Each phase of production of secondary conducting tissues (wood as well as bark) is followed by a phase of starch storing.

In the stem several false wood rings are produced during one year, while in the young branches each wood ring corresponds to one year.

The longitudinal growth and cork formation on the branches of the year are particularly pronounced during spring (March-May).

Cambium cells swell in a characteristic way before they start dividing and collapse during the resting periods, thickening evidently their walls, so that in winter it is difficult to distinguish a cambial cell from a parenchimatic one.

The behaviour of the vegetative growth of the olive tree in Messina is discussed on the base of the climatic characters of the region.     

Seasonal Changes of the Metabolites in the Leaves, Bark and Xylem Tissues of Olive Tree (Olea europaea. L) I. Nitrogenous Compounds

The changes of total nitrogen and free amino compounds wereexamined in leaves, bark and xylem tissues of olive tree duringa complete annual cycle. In leaves total nitrogen decreasedin spring/early-summer but reached the highest level in autumn-winterperiods. In bark tissues total nitrogen fluctuations were mainlyrelated to the periodical cambial activities. In xylem tissuestotal nitrogen was detected in low levels with no fluctuationsover the year. The free amino compounds in leaves showed seasonalpeaks during the spring and autumn stem elongations, while aconsiderable restriction of their levels was detected in summer.Glutamate, aspartate, proline, alanine, serine and -aminobutyrateare the predominant amino acids detected in leaves. Glutamateand aspartate increased considerably in spring and autumn periodsbut disappeared in summer. Proline remained almost unchangedover the year. Alanine, in addition to the spring and autumnfluctuations, also fluctuated in summer, -aminobutyrate wasdetected at highest levels in winter. In bark tissues, the patternof fluctuations and the composition of the free amino compoundswere similar to those of leaves. In xylem tissues free aminocompounds were detected in high levels over the year exceptfor the drops in spring and summer cambial activity periods.Xylem was the most important reservoir for the readily transportablesoluble nitrogen being accumulated during the maturing of theproduced new xylem. Arginine, glutamine, alanine, glutamate,aspartate, serine, -aminobutyrate and proline are the most prevalentamino compounds in xylem. Arginine and glutamine showed extensiveinterchanges. Arginine increased in autumn while glutamine andalanine showed low levels in the same period. Olive tree (Olea europaea L), amino acids     

Seasonal regulations of resting metabolic rate and thermogenesis in striped hamster (Cricetulus barabensis)

(1)

Resting metabolic rate (RMR), nonshivering thermogenesis (NST) and mitochondria cytochrome c oxydase (COX) activity of brown adipose tissue (BAT), as well as weight of skin and fur were measured in striped hamsters (Cricetulus barabensis) that were live-trapped in the summer, autumn, winter and spring.     

PENG Fang-Ren银杏营养贮藏蛋白质的亚细胞定位(英文)

在电子显微镜下,对银杏(GinkgobilobaL.)枝条营养贮藏蛋白质的超微结构特征及在亚细胞水平的定位进行了系统研究。结果表明:银杏营养贮藏蛋白质主要存在于韧皮薄壁细胞的液泡内。银杏韧皮薄壁细胞内的营养贮藏蛋白质在细胞质内合成,由内质网膨大的槽库、质膜内折或高尔基体小泡发育形成贮藏蛋白质的液泡。液泡蛋白质主要以不定形块状、絮状或颗粒状形态存在。贮藏蛋白质在整个越冬期一直保持高含量,直到翌年春季萌芽时,贮藏蛋白质迅速转移再利用。随着新梢的生长,到了夏末秋初,又重新开始积累贮藏蛋白质。     

Seasonal Variation in the Ultrastructure of the Cambium in New Zealand Grown Pinus radiata D. Don

Examination of Pinus radiata cambium from trees growing in thecentral North Island of New Zealand has revealed that many ofthe structural changes occurring in other tree species at theonset or cessation of cambial activity are not found in thisspecies. The winter cambium bears a closer resemblance to thesummer cambium than it does to the winter cambium of any otherangiosperm or gymnosperm described in the literature. In particularthere is little change in vacuolar structure, endoplasmic reticulumform, and dictyosomal activity during the year. The only changeswhich take place involve a slight increase in vacuole volume,storage of starch in vacuoles, and a decrease in numbers ofspherosomes during the summer. These observations confirm that,while the degree of cambial activity is reduced in the winter,complete dormancy is absent.     

Correct glycosylation,Golgi-processing,and targeting to protein bodies of the vacuolar protein phytohemagglutinin in transgenic tobacco

We used a heterologous system (transgenic Nicotiana tabacum L.) to investigate the processing, assembly and targeting of phytohemagglutinin (PHA), the lectin of the common bean, Phaseolus vulgaris L. In the bean, this glycoprotein accumulates in the protein bodies of the storage parenchyma cells in the cotyledons, and each polypeptide has a high-mannose glycan attached to Asn12 and a complex glycan on Asn60. The gene for PHA-L, dlec2, with 1200 basepairs (bp) 5 upstream and 1600 bp 3 downstream from the coding sequence was introduced into tobacco using Agrobacterium-mediated transformation (T. Voelker et al., 1987, EMBO J. 6, 3571–3577). Examination of thin sections of tobacco seeds by immunocytochemistry with antibodies against PHA showed that PHA-L accumulated in the amorphous matrix of the protein bodies in the embryo and endosperm. This localization was confirmed using a non-aqueous method to isolate the protein bodies from mature tobacco seeds. The biochemical analysis of tobacco PHA indicated that the signal peptide had been correctly removed, and that the polypeptides formed 6.4 S oligomers; tobacco PHA had a high-mannose glycan at Asn12 and a complex glycan at Asn60. The presence of the complex glycan shows that transport to the protein bodies was mediated by the Golgi complex. At seed maturity, a substantial portion of the PHA-L remained associated with the endoplasmic reticulum and the Golgi complex, as indicated by fractionation experiments using aqueous media and the presence of two high-mannose glycans on some of the polypeptides. Taken together, these data show that insertion of the nascent PHA into the endoplasmic reticulum, signal peptide processing, glycosylation, assembly into oligomers, glycan modification in the Golgi, and targeting of the protein occur faithfully in this heterologous system, although transport may not be as efficient as in bean cotyledons.Abbreviations Asn asparagine - Endo H endoglycosidase H - HPLC high-performance liquid chromatography - IgG immunoglobulin G - M_r relative molecular mass - PAGE polyacrylamide gel electrophoresis - PHA phytohemagglutinin - SDS sodium dodecylsulfate - TFMS trifluoromethanesulfonic acid     

Light and electron microscopical localization of concanavalin A lectin binding sites in rat epiphyseal chondrocytes

Summary Concanavalin A lectin binding sites have been detected within the cytoplasm of epiphyseal chondrocytes. Correlative light and electron microscopic results were obtained, indicating the presence of-d-mannose and/or -D-glucose residues detected by the lectin in the rough endoplasmic reticulum region. Quantitation of the electron microscopic cytochemical reaction also showed that the specific labelling was almost exclusively localized in the lumen of endoplasmic reticulum cisternae. No significant staining was found in other membrane compartments or extracellular matrix. This labelling pattern could be considered as the cytochemical evidence ofN-glycosylation processes occurring during the biosynthesis of cartilage extracellular matrix components by chondrocytes.     

Spatial and temporal variations of Penilia avirostris and Evadne tergestina (Crustacea,Branchiopoda) in a Tropical Bay,Brazil

We analysed monthly samples collected in Guanabara Bay, with a conical net of 200 m mesh during 1985. The bay was divided into three areas: an outer region (area A), influenced by oceanic waters; an inner region (area C), influenced by fluvial inflow; and a transition region (area B) with intermediate features. Penilia avirostris and Evadne tergestina were observed in the three areas, with greatest densities, however, in the outermost region, which had the highest salinities and lowest temperatures. Penilia avirostris was more abundant in summer (March), a period with the greatest relative densities of nanoplankton. Evadne tergestina was also abundant in summer, but its peak fell in November, a period with a relative increase in microphytoplankton density in the bay. The two species disappeared in winter: Penilia avirostris was absent from May to August, whereas Evadne tergestina disappeared in August and September.     

Seasonal variation in the activity of the Leydig cells in Egyptian Nubian goat (Zaraibi) bucks

Ten Egyptian Nubian goat bucks were used to evaluate the effect of season on testicular hormonal activity and ultrastructure. Parameters were recorded for 7 consecutive weeks in the middle of the four seasons, with blood samples being collected weekly. At the end of each of these seasons, testicular biopsies were obtained surgically for histological and cytological studies. Season had a significant effect on plasma testosterone concentration, being at its lowest level (P < 0.01) during winter and spring (1.2 and 2.6 ng/ml, respectively), while at its highest during summer (10 ng/ml). The effect of season on plasma LH concentration was higher (P < 0.01) in autumn (2.9 mIU/ml) and less in spring and summer (0.4 mIU/ml). Season of the year influenced the percentage of sectional tissue area occupied by the seminiferous tubules and interstitial tissue. Seminiferous tubules occupied the majority of the testicular tissue during winter (76.6%), with the least being occupied during spring (49.8%). The thickness of the seminiferous tubules was maximal during autumn, followed by summer (53 and 36 μm, respectively). In summer the Leydig cells contained abundant smooth endoplasmic reticulum (sER), while some areas of the cytoplasm were occupied exclusively by tubular sER, arranged in parallel—indicating the highest activity of these cells. A characteristic multivesicular structure with numerous large lipid droplets and vacuoles was recorded in the Leydig cells during spring and winter, denoting low or even arrested activity of the cells. It could be concluded that season influences the activity of the Leydig cells of Egyptian Nubian bucks, and this is reflected by their ultrastructure and secretive activity.     

Seasonal variations in the immune system of the tench,Tinca tinca (Cyprinidae): proliferative response of lymphocytes induced by mitogens

The proliferation of tench lymphocytes induced by mitogens was studied during the four seasons of the year. Fish were maintained under natural conditions of photoperiod and temperature (mean ± SD: 12±2°C in winter, 22±3°C in spring, 30±3°C in summer and 21±3°C in autumn). Cultures were performed in vitro at 22°C in all seasons and the results were compared. Subsequently, in seasons with extreme water temperatures, cultures in vitro were performed at the same temperature as that of the water (12°C in winter and 30°C in summer) and the results were compared seasonally at the seasonal temperature, i.e. at 22°C in spring, 30°C in summer, 22°C in autumn and 12°C in winter. Phytohemagglutinin, concanavalin A, lipolisaccharide from E. coli and pokeweed mitogen were used as mitogens. Studies performed at 22°C as assay temperature in all seasons showed profound seasonal changes: while in spring, summer and autumn the mitogenic response of lymphocytes to phytohemagglutinin, concanavalin A, lipolisaccharide from E. coli and pokeweed mitogen was very low, during winter the results obtained were significantly higher. However, when the assays were performed at the corresponding seasonal temperature the differences were not as pronounced between the different seasons, and the mitogenic responses of lymphocytes were found to be the lowest during the winter and the highest during the summer with all mitogens used. This fact suggests that immunosuppression occurs in winter and an immunostimulation occurs in summer. However, the higher response found in winter when assaying at 22°C suggests that this property of lymphocytes needs an assay temperature higher than the in vivo temperature in order to observe accurate mitogenic responses.Abbreviations Con A concanavalin A - cpm counts per minute - LPS E. coli lipolisaccharide - MS222 tricainemethane sulphonate - PBS phosphate-buffered saline - PHA phytohemagglutinin - PWM pokeweed mitogen - SI stimulation index     

Subcellular localization of calcium in sporangiophores of Phycomyces blackesleeanus

The in situ localization of Ca²⁺ in stage I sporangiophores of the fungus Phycomyces blakesleeanus was achieved with the potassium pyroantimonate technique. Precipitates of calcium-antimonate were present in mitochondria, vacuoles, endoplasmic reticulum and adjacent cytoplasm, Golgi-like bodies, and nuclei but not cell walls. Material treated with the calcium chelator EGTA lacked these precipitates. The preferential localization of Ca²⁺ in mitochondria, endoplasmic reticulum and vacuoles suggests that these organelles modulate the level of this cation in sporangiophores of P. blakesleeanus.Abbreviations EGTA ethyleneglycol-bis-(-aminoethyl ether) N,N, tetraacetic acid     

Ultrastructure and histochemistry of Citrus limon (L.) stigma

Citrus limon has a wet stigma which can be divided in two zones: a glandular superficial one formed by papillae, and a non-glandular one formed by parenchymatic cells. The stigmatic exudate is produced by the papillae after the latter have reached their ultimate size. The papillae of the mature pistil are of varying size and composition. Both the unicellular and multicellular ones are present. The cells at the base of the papillae are rich in cytoplasm, whereas the tip cells are vacuolated. Histochemical analysis has shown that the exudate of Citrus is composed of lipids, polysaccharides, and proteins. Our results indicate that the lipidic component is produced and secreted first, followed by production and secretion of the polysaccharidic component. The lipidic component of the exudate is produced in the basal papillae cells and accumulates as droplets in dilated parts of the smooth endoplasmic reticulum (SER). Subsequently the lipid droplets are transported to the plasma membrane, and transferred by the latter into the cell walls. Then the exudate component is accumulated in the intercellular spaces and in the middle lamellar regions of the walls. Subsequently, the polysaccharidic component of the exudate is produced and secreted by the tip cells of the papillae.Abbreviations RER rough endoplasmic reticulum - SER smooth endoplasmic reticulum     

Changes in endoplasmic reticulum during spermiogenesis in the mouse

Summary Changes in the endoplasmic reticulum of mouse spermatids during spermiogenesis were examined by scanning electron microscopy, applying the OsO₄-DMSO-OsO₄ method, which permits 3-dimensional observation of cell organelles. At the same time, the endoplasmic reticulum was stained selectively by the Ur-Pb-Cu method, and 0.5 m-thick sections were prepared for observation by transmission electron microscopy. The results demonstrated stereoscopically the mode of disappearance of the endoplasmic reticulum. In spermatids of the early maturation phase, the endoplasmic reticulum was of uniform diameter, branched and anastomosed, forming a complicated three-dimensional network throughout the cytoplasm. A two-dimensional net was also noted to have formed just beneath the plasma membrane and about Sertoli cell processes invaginating the spermatid cytoplasm. As spermiogenesis progressed, the spread-out endoplasmic reticulum gradually aggregated to form a condensed, glomerulus-like structure consisting of a very thin endoplasmic reticulum connected to the surrounding endoplasmic reticulum. This structure corresponds to the so-called radial body. Thus, the endoplasmic reticulum may aggregate, condense, be transformed into a radial body, and be removed from the cytoplasm. The two-dimensional endoplasmic reticulum-net, just beneath the plasma membrane and surrounding processes of Sertoli cells, disappeared in spaces where the three-dimensional endoplasmic reticulum network was scarce. Both the two-dimensional endoplasmic reticulum-net structure and the three-dimensional endoplasmic reticulum network disappeared at the same time, indicating that they may be closely related.     

Changes in the pattern of phospholipid synthesis during the induction by cytokinin of cell division in soybean suspension cultures

A method is described for preparing fully viable, cytokinin-starved soybean (Glycine max (L.) Merr. cv. Acme) cells from a suspension-culture of callus tissue. The cells respond to kinetin treatment by re-initiating cell division. We present evidence, from the pattern of incorporation of ³²P-labelled inorganic phosphate into individual phospholipids during the first hour of this response, that the synthesis of phosphatidylinositol (PI) and of phosphatidic-acid head-groups is affected within 15 min. The polyphosphoinositide phosphatidylinositol 4-phosphate, but not phosphatidylinositol 4,5-bisphosphate, was detected in the tissue. The characteristics of cytokinin-induced PI synthesis in cytokinin-starved soybean cells appear to resemble the PI response of animal cells.Abbreviations DPG diphosphatidylglycerol - PA phosphatidic acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PG phosphatidylglycerol - PI phosphatidylinositol - PIP phosphatidylinositol 4-phosphate - PIP₂ phosphatidylinositol 4,5-bisphosphate - PS phosphatidylserine - Pi inorganic phosphate - TLC thin-layer chromatography     

Stereological study of gonadotropes in the frog,Rana pipiens,after GnRH stimulation in vitro

Summary Previous physiological results have indicated the existence of two releasable pools of gonadotropins in amphibian pituitaries: an acute releasable pool that appears independent of protein synthesis, and a storage pool involved in chronic release that depends on protein synthesis. To elucidate the ultrastructural localization of these pools and the morphological changes induced in gonadotrope cells after treatment with gonadotropin-releasing hormone, we carried out a morphometric study of immuno-identified gonadotrope cells using an in vitro superfusion system. Treatment with gonadotropin-releasing hormone induced a degranulation of small (110–255 nm) and medium (236–360 nm) secretory granules as well as hypertrophy of the endoplasmic reticulum and Golgi complex. Simultaneous incubation with gonadotropin-releasing hormone and cycloheximide inhibited the release of secretory granules although the endoplasmic reticulum and Golgi complex were hypertrophied. These morphological results strongly suggest: (1) that gonadotropin-releasing hormone induces degranulation and hypertrophy of the biosynthetic machinery in gonadotrope cells; and (2) that the activation of the endoplasmic reticulum and Golgi complex by stimulation with gonadotropin-releasing hormone is independent of protein synthesis, while the release of secretory granules is protein synthesis-dependent. In addition, the second or storage pool of gonadotropin is associated mainly with the small and medium secretory granules.     

Ontogeny of glyoxysomes in maturing and germinated cotton seeds—a morphometric analysis

Morphometric procedures were used with light and electron microscopy to examine glyoxysome number, volume, shape and distribution as well as mesophyll cell volume, in cotyledons of mature (50 d postanthesis), imbibed (5h) and germinated (24 and 37 h) cotton (Gossypium hirsutum L.) seeds. Additionally, activities of five glyoxysomal marker enzymes in cotyledon extracts were assayed at each of the above ages. Cell volume was determined from photomicrographs of Epon-embedded sections by the point-counting procedure. Analysis of variance showed that cell volume was not different among the tissue segments studied. Glyoxysomes were cytochemically stained for catalase (EC 1.11.1.6) activity with the 3,3-diaminobenzidine-tetrahydrochloride procedure. Analyses involving both phase and electron microscopy, and two separate sterologic calculations for determining the number of glyoxysomes per cell, indicate that glyoxysomes are numerous in mature seeds, persist through desiccation and imbibition, then increase dramatically in volume (seven fold) but not number (a maximum of 1.5-fold), when enzyme activities increase two to six times (depending on the enzyme). During the entire period of increase in glyoxysomal enzyme activities, no ultrastructural evidence was found for glyoxysome formation or destruction. Our data, in contrast to some proposals in the literature, indicate that cottonseed glyoxysomes form during seed maturation, then develop following seed imbibition into pleomorphic organelles by posttranslational accumulation of proteins from the cytosol and transfer of membrane components probably from the endoplasmic reticulum.Abbreviations DAB 3,3-diaminobenzidine tetrahydrochloride - DPA days postanthesis - ER endoplasmic reticulum     

Localization of mRNA for pea legumin:In situ hybridization using a biotinylated cDNA probe

Summary In situ hybridization has been used to locate mRNA, for the storage protein legumin, in cotyledon storage parenchyma tissue of developing pea (Pisum sativum L.) seeds. The mRNA was hybridized with a biotinylated probe of cDNA in pBR 322 and subsequently located by avidin conjugates. Avidin-rhodamine was used for fluorescence microscopy localization at a tissue/cellular level and avidin-peroxidase (with DAB) and avidin-ferritin compared for localization at an ultrastructural level. Specific fluorescence associated with avidin-rhodamine was distributed unevenly throughout the cytosol but the cell walls, starch grains, vacuoles and protein deposits were unstained. The sizes and distribution of the regions of higher labeling within the cytosol suggest an association with elements of the endomembrane system. Following DAB reaction of the specifically localized avidin-peroxidase most, although not all, stain product was associated with the endoplasmic reticulum. The ER-associated reaction product was also accumulated within the ER lumen.Avidin-ferritin was also localized both in the cytosol and in association with the endoplasmic reticulum, although was less readily visualized in cells with a conventional ultrastructural appearance.Localization of avidin-ferritin was more readily visualized in cells which had undergone some limited structural damage during specimen preparation. In such cases ferritin was also shown to be specifically associated with the transition vesicles and trans-face peripheral vesicles of some dictyosomes.     

多尺度下宝鸡地区干旱动态格局演变及其与植被覆盖的关系

利用宝鸡地区11个气象站点1974—2013年逐月气温和降水量数据,基于标准化降水蒸散指数(SPEI),结合土地利用/覆盖数据,从干旱发生频率、发生强度及与植被NDVI相关性等角度,探讨了近40年来干旱时空变化格局及其对植被覆盖的响应。结果显示:宝鸡地区年均SPEI指数以-2.50%/a的速度下降,干旱趋势明显增强。自20世纪末以来,全区年均干旱指数呈明显的上升趋势,以2007—2010年增大趋势最为显著(超过0.05临界线);近40年来,春季干旱发生频率达60%及以上的有21 a,达90%以上的有9a。1981—1986年起伏变化最为剧烈。秋季平均干旱发生频率为46.29%,为春、夏、秋、冬4个季节中最低。从干旱发生强度来看,全区年际、四季及月6个时间尺度上干旱强度高、低值区域分布均比较集中;秋季强度最弱的区域面积表现最高,占总面积的75.47%。干旱发生最严重的是春季,占总面积的11.90%。全区干旱与植被覆盖相关性均表现较好(均通过0.05显著性水平检验),林地、草地负相关性最为显著(除夏季),夏季、秋季、月尺度上,耕地、水域、城乡地区干旱与植被覆盖的相关性与土地利用类型无关。