Transverse Vein Differentiation Associated with Gas Space Formation--Fate of the Middle Cell Layer in Leaf Sheath Development of Rice

In monocotyledons, the leaf vascular network consists of a hierarchicalsequence of vertical vascular bundles and numerous transverseveins that interconnect adjacent vertical veins. In the leafsheath of these species, especially grasses, lysigenous gascavities (gas spaces) are developed into intervascular spacesand provide a gas conducting system to non-aerial parts underflooded conditions. The spatial relationship between gas spaceformation and transverse vein differentiation was investigatedusing the leaf sheath of rice (Oryza sativa L.). Histochemicalobservation showed that patterns of differentiation of the transversevein are distinct from those of vertical vascular bundles. Onthe other hand, gas spaces are formed through the processesof cell death (collapse). Both events are initiated at a specificcell position in the middle layers of the leaf sheath, fromwhich the vascular system of the leaf is derived; this indicatesthat differentiation of transverse veins is associated withgas space formation. The cell-to-cell movement of fluoresceinisothiocyanate-conjugated dextran injected into middle layercells coincided with the area where cell collapse occurred,indicating a close relationship between the middle and adaxialcell layers, but not abaxial cell layers. A uniform cell numberbetween each transverse vein in the leaf sheath suggested theinvolvement of spatial regulation in transverse vein formationregardless of clonal history at the later stage of leaf veincanalization. Copyright 2000 Annals of Botany Company Cell collapse, leaf development, middle cell layer, microinjection, Oryza sativa L., rice, programmed cell death.     

Establishment of a Murine Model for Metastasis of Cytokine-Producing Tumor to the Brain

The A375 cell line, derived from human malignant melanoma, has characteristics of interleukin-6 (IL-6) production. By using this cell line, we have investigated a murine metastasis model of IL-6-producing tumors to the brain by injecting A375 cells directly into the left cardiac ventricle. Nude mice were anesthetized with intraperitoneal injection of pentobarbital sodium. Next, A375 cells suspended in phosphate-buffered saline (PBS) were injected into the left cardiac ventricle of mice. An intracardiac injection of 10⁵ cells developed tumor colonies in the brain after 4 to 6 weeks. Metastatic cells were found in every lobe of the brain. An immunocytochemical study revealed IL-6 production by A375 cells at the metastatic sites in the brain. By the transfection of genes encoding proteins into A375 cells, a novel model of protein expression in the brain in vivo could be constructed. Our system does not require great skill. Our experimental model will facilitate future studies of the local effects of proteins in the brain.     

Some properties of shikimate: NADP oxidoreductase produced in sweet potato root tissue after slicing

The activity of shikimate: NADP oxidoreductase [EC 1. 1. 1.25] in sweet potato root tissue increased soon after slicing.Enzyme preparations obtained from both sliced tissue and fromfresh tissue probably contained a single enzyme component, andthey showed identical chromatographical behaviour. K_m values of the enzyme for NADP and shikimate were 1.0x10^–4Mand 1.3 x 10^–3M, respectively. Enzyme activity was potentlyinhibited by SH-inhibitors such as p-chloromercuribenzoate andoxidized glutathione. Enzyme activity was affected neither by mononucleotides suchas ATP, ADP and AMP, divalent cations, Mg⁺⁺, Ca⁺⁺ and Mn⁺⁺,nor by metabolites such as tryptophan, phenylalanine, tyrosineand t-cinnamic acid which are involved in aromatic compoundsyntheses. The enzyme rapidly lost its activity. This inactivation reactionshowed a time course consisting of two steps of the first-orderreaction. The inactivated enzyme preparation was not reactivatedby thiol compounds such as cysteine, 2-mercaptoethanol and glutathione,although these reagents, to a certain extent, protected theenzyme from inactivation. The results suggest that denaturationof the enzyme protein was involved in inactivation of the enzyme. ¹Part 74 of the phytopathological chemistry of sweet potatowith black rot and injury. ²Present address: Department of Biology, Faculty of Science,Tokyo Metropolitan University, Setagaya-ku, Tokyo. (Received August 5, 1968; )     

The Participation of Speract in the Acrosome Reaction of Hemicentrotus pulcherrimus

A speract-free macromolecular fraction was prepared from the egg jelly of the sea urchin Hemicentrotus pulcherrimus by gel filtration and tested for ability to induce the acrosome reaction in H. pulcherrimus spermatozoa with or without exogenously added synthetic speract. The macromolecular fraction without speract showed only about half the activity of the original unfractionated jelly for induction of the acrosome reaction. The rates of the acrosome reaction induced by the fraction with speract were comparable with those induced by the unfractionated jelly at all pHs tested. Speract itself, however, did not induce the acrosome reaction in the absence of the macromolecular fraction of jelly. The acrosome reaction was associated with incerase of the cyclic AMP concentration in sperm cells, the extent of incerase depending on the concentration of the macromolecular fraction. Addition of speract to the fraction enhanced both induction of the acrosome reaction and increase in the cyclic AMP concentration induced by the fraction. These results suggest that a major factor(s) responsible for the acrosome reaction is a macromolecular component(s) of the jelly and that speract promotes the reaction as a co-factor.     

Development of Second Generation Merozoites of Leucocytozoon caulleryi In Vitro

Sporozoites of Leucocytozoon caulleryi were inoculated into specific-pathogen-free (SPF) chickens intravenously. Fourteen days after inoculation, the infected blood, parasitized with second generation merozoites, was collected from the chickens. The blood was then suspended in SPF chicken serum or RPMI-1640 culture medium supplemented with 20% SPF chicken serum in petri dishes, which were cultured at 37°C or 41°C in a humidified atmosphere of 5% CO₂. Second generation merozoites in parasitized, immature erythrocytes developed continuously through several substages and finally became micro- and macro-gametocytes after 6 days of incubation.     

Phylogeography of lions (Panthera leo ssp.) reveals three distinct taxa and a late Pleistocene reduction in genetic diversity

Lions were the most widespread carnivores in the late Pleistocene, ranging from southern Africa to the southern USA, but little is known about the evolutionary relationships among these Pleistocene populations or the dynamics that led to their extinction. Using ancient DNA techniques, we obtained mitochondrial sequences from 52 individuals sampled across the present and former range of lions. Phylogenetic analysis revealed three distinct clusters: (i) modern lions, Panthera leo ; (ii) extinct Pleistocene cave lions, which formed a homogeneous population extending from Europe across Beringia (Siberia, Alaska and western Canada); and (iii) extinct American lions, which formed a separate population south of the Pleistocene ice sheets. The American lion appears to have become genetically isolated around 340 000 years ago, despite the apparent lack of significant barriers to gene flow with Beringian populations through much of the late Pleistocene. We found potential evidence of a severe population bottleneck in the cave lion during the previous interstadial, sometime after 48 000 years, adding to evidence from bison, mammoths, horses and brown bears that megafaunal populations underwent major genetic alterations throughout the last interstadial, potentially presaging the processes involved in the subsequent end-Pleistocene mass extinctions.     

黄河三角洲南部潮间带沉积环境对底栖叶绿素a分布特征的影响

2007年9月、2008年4月和2008年7月,在东营黄河三角洲南部潮间带进行了表层沉积物叶绿素a含量和粒径组成调查。结果表明:1)近岸区域叶绿素a含量由北向南逐渐增高,秋季和夏季叶绿素a含量分别为0.426～4.749μg.g-1(平均值为1.591μg.g-1)和0.307～5.195μg.g-1(平均值为1.832μg.g-1),由潮上线到潮下线呈下降趋势;春季叶绿素a含量为0.623～8.734μg.g-1(平均值为2.542μg.g-1),由潮上线到潮下线呈上升趋势;2)沉积物类型较为单一,由粉砂、砂质粉砂和粉砂质砂组成,中值粒径变化范围为0.025～0.081mm(平均中值粒径为0.045mm),由潮上线到潮下线逐渐增大;3)2007年9月叶绿素a分布主要受沉积物粒度的影响,与细颗粒物质含量、中值粒径的大小及各粒径沉积物含量极显著性相关(P0.01);2007年9月与2008年7月叶绿素a含量与细颗粒物质含量呈正比关系,在砂质底质中含量较低,在粘土质底质中含量较高;4)底栖叶绿素a与高程呈负相关,表明松软、平整、稳定的沉积物表面有利于底栖微型藻类的生长和繁衍。     

Development of microsatellite markers in the noxious red tide‐causing algae Heterosigma akashiwo (Raphidophyceae)

We isolated 13 polymorphic microsatellites from the noxious red tide‐causing algae, Heterosigma akashiwo. These loci provide a class of highly variable genetic markers as the number of alleles ranged from three to 12 and the observed and expected heterozygosities ranged from 0.286 to 0.926 and from 0.314 to 0.888, respectively. We consider these loci potentially useful for detailing the genetic structure and gene flow among H. akashiwo populations.     

Genetic structuring and transfer of marine dinoflagellate Cochlodinium polykrikoides in Japanese and Korean coastal waters revealed by microsatellites

To determine the process of population expansion and ascertain the origin of the Sea of Japan population, in a noxious red tide forming dinoflagellate Cochlodinium polykrikoides , 13 samples, isolated from 11 different localities in Japanese and Korean coasts, were analysed using 10 polymorphic microsatellites. Analyses by nonmetric multidimensional scaling plots of pairwise F _ST, global amova , and genetic admixture analysis identified three clusters — the Sea of Japan populations, Yatsushiro Sea (Kumamoto Pref.) populations, and other populations — indicating genetic structuring of the 13 samples into three distinct populations. In the proportion of shared alleles by pairwise individuals ( P _SAxy) analyses between the Sea of Japan and the other samples, P _SAxy was extremely low compared with that among the Sea of Japan or among other samples, indicating that a large genetic barrier has occurred between the populations. No significant relationship of isolation-by-distance patterns and almost no genetic distance were detected between pairwise samples of the Sea of Japan, although there is a maximal distance of > 600 km between samples. In addition, P _SAxy data among the samples were extremely high compared with those among other samples, clearly showing that a large-scale transfer from west to east has occurred via the Tushima Warm Current. In the P _SAxy data of the Seto Inland Sea and Pacific samples, individuals showing relatively high P _SAxy were concentrated in the three areas of Nagasaki, Harima, and Mie, suggesting that frequent transfer may have occurred by human-assisted dispersal, although Nagasaki and Mie are separated by a distance of approximately 700 km.     

HORMONES CONTROLLING COLLAGEN SYNTHESIS DURING METAMORPHOSIS IN THE THIGH BONE OF THE TADPOLE OF RANA CATESBEIANA

The rate of ¹⁴C-proline incorporation into collagen in the thigh bone of the Rana catesbeiana tadpole was determined in vitro. Intraperitoneal injection of bovine prolactin caused an increase in the rate of collagen synthesis during the premetamorphic stages (stages 12–16) and the early metamorphic stage (stage 18), but it exerted no effect on collagen synthesis in the metamorphic stages (stages 20–25). On the other hand, injection of growth hormone stimulated the rate of collagen synthesis in the metamorphic stages and caused a slight increase in the premetamorphic stages. When a tadpole in the early premetamorphic stages (stages 12–14) was kept in 5 × 10₋₈ M thyroxine solution for several days, the rate of collagen synthesis became higher than that in the bone of the control animal. The rate of collagen synthesis was not enhanced by prolactin in the thyroxine-treated tadpole, but was stimulated by growth hormone, even when the thyroxine-treated animal remained in the premetamorphic stages. With the treatment of the tadpole by thyroxine, prolactin-sensitivity seems to be reduced, and growth hormone-sensitivity becomes apparent.