Pollination by fungus gnats in four species of the genus Mitella (Saxifragaceae)

The first example of pollination by fungus gnats in the eudicots is reported. The genus Mitella (Saxifragales) is characteristically produces minute, inconspicuous, mostly dull-coloured flowers with linear, sometimes pinnately branched, petals. To understand the function of these characteristic flowers, we studied the pollination biology of four Mitella species with different floral traits and different sexual expression: dioecious M. acerina , gynodioecious M. furusei var. subramosa , and hermaphroditic M. stylosa var. makinoi and M. integripetala. Flower-bagging experiments showed that wind pollination did not occur in the dioecious and gynodioecious species. Two years of observations of flower visitors at six study sites in Japan revealed that the principal pollinators of all four Mitella were specific species of fungus gnats (Mycetophilidae), which landed on the flowers with their long spiny legs settling on the petals. Characteristically, numerous pollen grains were attached to the fungus gnats in specific locations on the body. Although, on average, 1.3–2.6 fungus gnats visited each inflorescence per day, the fruit set of both bisexual and female flowers exceeded 63%. These results suggest that fungus gnats are highly efficient pollinators of Mitella spp., and that Mitella flowers are morphologically adapted to pollination by fungus gnats.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 449–460.     

Reproductive and Pollination Biology of Magnolia and its Allied Genera (Magnoliaceae). I. Floral Volatiles of Several Magnolia and Michelia Species and their Roles in Attracting Insects

Abstract Volatile substances emitted from the flowers of eight Magnolia taxa ( M. sieboldii ssp. japonica, M. praecocissima var. praecocissima and var. borealis, M. tomentosa, M. salicifolia, M. obovata, M. denudata, and M. grandiflora ) and one Michelia species ( M. compressa ) (Magnoliaceae) were examined and identified using GC-MS. Volatile substances of these Magnolia and Michelia species consist primarily of monoterpenoids and sesquiterpenoids produced by the mevalonate pathway, acetogenins by the acetate-malonate pathway, and phenyl-propanoids by the shikimate pathway. These Magnolia and Michelia species all possessed various combinations of volatile monoterpenoids, acetogenins, and phenylpropanoids, except for Magnolia obovata , which emitted primarily sesquiterpenoids. Free amino acids in pollen of 12 Magnolia and one Liriodendron species were also analyzed, and their value as food sources for pollinators evaluated.
Pollinators visiting the flowers of five Magnolia species were collected in their native sites and identified. Their behaviors and roles as pollinating agents were assessed.     

The Cell Cycle and Cell Population Kinetics in the Programmed Cell Death in the Limb-Buds of Normal and 5-Bromodeoxyuridine- Treated Chick Embryos

The cell cycle and cell population kinetics have been analyzed in the interdigital regions of chick limb-buds during the course of programmed cell death both in normal and the 5-bromodeoxyuridine (BrdU)-treated embryos. Our previous study has shown that a single administration of BrdU at day 6 1/3 inhibited the programmed cell death occurring in normal development of limb-buds.
Pulse- as well as continuous labelings with tritiated thymidine (³H-TdR) were used. The results obtained from the analyses made on both normal and experimental embryos have demonstrated the presence of a particular DNA-synthetic period, around day 6 1/3, closely related to the programmed death occurring on day 7 1/3. In normal embryos, new cell populations, which did not belong to any phases of normal cell cycle, made their appearances in the process of programmed cell death. A possible correlation between programmed cell death and the cell cycle has been discussed in relation to the morphogenesis of limbs in both normal and BrdU-treated embryos.     

Unique Pattern of Gene Expression in the Erythroid Precursor Cells

Erythroid cells were fractionated by preformed Percoll density gradient from livers of 12.5 day old mouse fetuses. With combination of lysing of mature erythroid cells, the CFU-E (colony forming unit of erythroid) was enriched as high as 30% pure. The mRNA levels of the rt-genes previously cloned as genes expressed in the reticulocytes are estimated in the fractionated erythroid cells. These rt-genes show a drastic change in expression during erythroid differentiation; Their expression was not detectable at the CFU-E cell stage. But it reached to maximum at the polychromatic erythroblast (stage I) and then decreases with maturation. The result suggests that mRNA synthesis of these rt-genes may be induced after the stimulation of erythropoietin.     

Reactivity of a Monoclonal Antibody Raised against Human Leukemic Cells to Embryonic and Adult Tissues of the Mouse and Teratocarcinomas

C-9-1, a monoclonal IgM antibody raised against human null cell acute lymphocytic leukemia cells reacted with restricted regions of embryonic and adult tissues of the mouse. The antigen positive sites in the embryos included embryonic ectoderm, visceral endoderm, trophoblastic cells invading the maternal decidua of 5∼7-day embryos, primordial germ cells of 10∼12-day embryos, epithelium of nasal chamber, the bronchus, Mullerian duct, epididymis and bladder of 12∼17-day embryos. In the adult mice, C-9-1 antigen was detected in renal tubules, a part of stomach, bladder, endometrium and epididymal sperm. Embryonal carcinoma cells, but not endodermal cells of teratocarcinoma expressed the antigen. Thus, C-9-1 antigen showed distribution similar to SSEA-1. However, C-9-1 antigen was not detected in preimplantation embryos, nor in oviduct, both of which are positive for SSEA-1.     

Reactivation of the Hill reaction of Tris-washed chloroplasts

The Hill reaction of chloroplasts was inhibited by washing themwith 0.8 M Tris buffer. This inhibition was further promotedby adding ferricyanide in the washing medium. When a reducingreagent, such as the 2,6-dichlorophenol indophenol (DCPIP)-ascorbatesystem or the hydroquinone (HQJ-ascorbate system, had been addedto the Tris buffer, Hill reaction activity was unaffected. Hill reaction activity of Tris-washed chloroplasts recoveredup to 70% of the initial level by re-washing the chloroplastswith a preparation medium containing theabove reducing reagents. Photobleaching of carotenoid and chlorophyll is characteristicof Tris-washed chloroplasts. However, reactivated chloroplastsshowed no photobleaching as in the case with intact chloroplasts. (Received April 20, 1970; )     

DIRECT ISOLATION OF NATIVE THIN FILAMENTS FROM EMBRYONIC MUSCLE CELLS*

A method is presented for the release of “native” thin filaments from 13-day old embryonic chick muscle without tryptic digestion or desoxycholate (DOC) solubilization of Z bands. The isolated filaments were 50–60 Å in diameter, of variable length, and formed “arrowhead-like” complexes with heavy meromyosin (HMM). In addition, the filaments interacted with purified myosin to form actomyosin as effectively as action extracted from an acetone powder of muscle. The Mg⁺⁺-dependent ATPase activity and extent of superprecipitation of the synthetic actomyosin required a low concentration of Ca⁺⁺, strongly suggesting the presence of troponin and tropomyosin on the thin filaments isolated from muscle at this stage of embryogenesis. The native thin filaments were more sensitive to trypsin than synthetic F-actin prepared from an acetone powder based on measurements of flow birefrengence, viscosity and the ability to activate myosin ATPase.     

Non-cyclic photophosphorylation by spinach grana treated with 0.8 M Tris buffer

1. Photophosphorylation was measured with spinach grana sampleswashed by 0.8 M Tris buffer at pH 8.0, which no longer catalyzedthe ferricyanide and NADP HILL reactions with water as the electrondonor. The photophosphorylation with the reaction mixture containing2 10^–4 M 2,6-dichlorophenol indophenol (DCPI) plus above2 10^–3 M ascorbate as the electron donor system insteadof water under anaerobic conditions was, in the most part, dependenton the addition of both PPNR (a nonheme iron protein requisitefor photosynthetic pyridine nucleotide reduction ; spinach ferredoxin)and NADP as the electron acceptor system. However, when ascorbateconcentration only was lowered to 2 10^–4 M, the entirephotophosphorylation proceeded, even in the absence of the electronacceptor system. 2. When the NADP added in the reaction mixture had been reducedby glucose-6-phosphate and glucose-6-phosphate dehydrogenasebefore illumination, the photophosphorylation with 2 10^–4M DCPI plus 6.7 10^–3 M ascorbate decreased to aboutthe same rate as that obtained without NADP. 3. The time course for photophosphorylation in the presenceof NADP was consistent with the time course for the photoreductionof NADP: On the complete reduction of NADP, the photophosphorylationstopped. 4. In the presence of 6.7 10^–3 M dichloropheny 1.1,1-dimethylureaor 3 10^–4 M o-phenanthroline, non-cyclic photophosphorylationwith 2 10^–4 M DCPI plus 6.7 10^–3 M ascorbateas the electron donor system decreased to about half that ofthe control, and the remaining activities were hardly affectedeven at higher concentrations of both inhibitors. The P/2e^–ratios of non-cyclic photophosphorylation in the absence andpresence of ophenanthroline were 0.74 and 0.48, respectively. ¹Present address: Department of Biology, University of California,San Diego, La Jolla, California 92037, U. S. A.     

Neurosecretion and Salt and Water Balance in the Annelida and Crustacea

The possible role of the neurosecretory system in regulationof salt and water has been studied in the annelids and crustaceans.In the earthworm, Lumbricus terrestris, a brain factor influences'the salt and osmotic concentration of the blood and coelomicfluid. Removal of the brain results in the increase of waterinflux with a decrease in the salt and osmotic concentrationsof the body fluids. The decreases in salt and osmotic concentrationscan be prevented by the implantation of the brain or the injectionof brain homogenates. In the freshwater crayfish, Procambarus clarkii, a factor, presumablysecreted in the brain and released in the eyestalk, seems tomaintain the normal permeability of the body surfaces to water.Eyestalk removal, which eliminates the release site, resultsin the increased influx of water with a decrease in the saltconcentration of the blood. A brain factor also seems to beinvolved in maintaining the sodium and osmotic concentrationsof the blood. In the semi-terrestrial grapsid crab, Metopograpsus messor,the thoracic ganglion, under the control of an eyestalk element,secretes a factor involved in increasing the permeability ofthe body surfaces to water. The removal of the eyestalks, theimplantation of the thoracic ganglion, or the injection of extractsof thoracic ganglia, results in changes in the osmotic concentrationof the blood tending toward that of the medium. In all threespecies studied, the neuroendocrine factors seem to be involvedprimarily in the regulation of the permeability of the bodysurfaces to water.     

POLYOL ACCUMULATION IN IN VITRO SYSTEMS OF BOMBYX EGGS

The mechanism of polyol accumulation in diapausing Bombyx eggs, conversion of [6-¹⁴C] glucose-6-phosphate into polyols and other neutral sugars was investigated in in vitro reaction systems. When a crude homogenate or a press juice of the eggs was incubated with [6-¹⁴C]glucose-6-P, the labelled trehalose, sorbitol and glycerol accumulated in the reaction mixture. In the press juice incubation system of developing eggs at day 1, ¹⁴C-sorbitol was detected in appreciable amounts, but it decreased rapidly with the development of the embryos. When the press juice was prepared from eggs in diapause, the formation of ¹⁴C-sorbitol was 3–5 times greater in eggs at early stages (day 2 to day 4) than in developing eggs.