Polymorphism of axillary bud galls induced by Rhopalomyia longitubifex (Diptera: Cecidomyiidae) on Artemisia princeps and A. montana (Asteraceae) in Japan and Korea, with designation of new synonyms

In the past, Rhopalomyia longitubifex, Rhopalomyia shinjii, and Rhopalomyia sp. (Diptera: Cecidomyiidae) have been regarded as independent species based on differences in the sizes and shapes of axillary bud galls induced on Artemisia montana (Asteraceae) in Japan and A. princeps in Japan and Korea. However, comparison of morphological features and molecular sequencing data indicate that these Rhopalomyia gall midges are identical and that the differences in gall shape are polymorphisms, although the measurements of gall height and diameter overlap slightly. This finding suggests that although galls have frequently been regarded as extensions of the phenotype of a species, differences in gall shape may not always be reliable for identifying gall‐inducing cecidomyiids. The older name, R. longitubifex, is applied to these gall midges, and the names that were applied to this species on later occasions are revised or synonymized. The mature and immature stages of R. longitubifex are redescribed and information on the distribution, host range, and gall size of this species is provided. We also discuss the role of gall polymorphism in the early stages of speciation.     

Resurrection of Rabdophaga salicivora Shinji (Diptera: Cecidomyiidae), a Japanese gall midge formerly misidentified as a North American species, Rabdophaga rigidae (Osten Sacken), with observations on the phylogenetic relationships of its populations in Japan and the Russian Far East

Intercontinental biotic connections between Eurasia and North America are common in many gall midge genera (Diptera: Cecidomyiidae), but only a few species have been recorded from both continents. In Japan, four gall midge species had been previously considered to be identical to North American species, but three of these cases have already been disproved. We examined the remaining species, Rabdophaga rigidae, which had been originally described from Japan as Rabdophaga salicivora in 1938, later recorded from the Russian Far East in 1967, and synonymized with a North American species, R. rigidae, in 1982. Morphological features and partial sequence data of the mtDNA cytochrome oxidase subunit I (COI) region suggested that the Japanese species is a distinct species and is identical to the species recorded from the Russian Far East. We therefore apply the original name, R. salicivora, to the Japanese and the Russian species. In addition, on the basis of a molecular phylogenetic analysis, we conclude that R. salicivora possibly came to the Japanese Archipelago through the Korean Peninsula and established itself first in the southern parts of Japan. Then, it expanded its distribution range to northern parts of Honshu, but could not reach Hokkaido, probably because of the Tsugaru Strait between Honshu and Hokkaido.     

Ommochrome Deficiency in an Albino Strain of a Terrestrial Isopod,Armadillidium vulgare

In order to clarify the cause of ommochrome deficiency in an albino strain of the terrestrial isopod, Armadillidium vulgare, levels of xanthom-matin, 3-hydroxykynurenine, 3-hydroxyanthranilic acid and tryptophan in whole body extracts of the albino and the wild type individuals were determined together with enzyme activities of kynurenine-3-hydroxylase, kynureninase and tryptophan-2,3-dioxygenase. Xanthommatin could not be detected in the albinos. The levels of 3-hydroxykynurenine and 3-hydroxyanthranilic acid were determined by high-performance liquid chro-matography (HPLC) with electrochemical detection and were markedly low in the albinos compared with the wild type individuals. In contrast to those, the tryptophan levels determined by HPLC with fluorescence detection did not differ significantly between the two phenotypes. In the albino A. vulgare, kynurenine-3-hydroxylase activity was lower and kynureninase activity was higher than in the wild type, although the differences were not statistically significant. Tryptophan-2,3-dioxygenase activity in the albinos was less than 10% that in the wild type. Thus, ommochrome deficiency in the albino A. vulgare is considered to be caused by the extremely low activity of tryptophan-2,3-dioxygenase.     

INDUCTION OF CELLULAR PROCESSES CONTAINING COLLAGENASE AND RETINOID BY INTEGRIN-BINDING TO INTERSTITIAL COLLAGEN IN HEPATIC STELLATE CELL CULTURE

Cultered hepatic stellate cells were induced to elongate long, multipolar cellular processes by interstitial collagen gel used as a substratum, as compared to flattened or round cell shapes on polystyrene surface or on Matrigel containing the basement membrane components, respectively. The process induction was inhibited by several reagents as follows: (1) anti-integrin α2 antibody; (2) an oligopeptide, DGEA, an integrin-binding sequence in type I collagen molecule; (3) wortmannin, a phosphatidylinositol 3-kinase inhibitor. Protein tyrosine phosphorylation was enhanced throughout cells including cellular processes by culturing on type I collagen gel. Dual fluorescence staining showed that the core of the processes contained microtubules, whereas the periphery of the processes comprised fibrillar actin. Thus, the process extension was found to depend on integrin-binding to type I collagen fibres, followed by signal transduction and cytoskeleton assembly. The cellular processes included interstitial collagenase and vitamin A-containing lipid droplets. The lipid droplets and vitamin A-autofluorescence were increased by retinyl acetate addition to the culture medium, suggesting an important role of processes in hepatic stellate cell function.     

The Effects of Ethanol on the Germination of Seeds of Japonica and Indica Rice (Oryza sativa L.) under Anaerobic and Aerobic Conditions

An examination was made of the effects of ethanol at 0.2–6.0%(v/v) on the germination, under aerobic conditions, of intactand dehusked seeds of indica rice (cv. Assam IV), which hadbeen harvested 14, 21 and 28 d after anthesis, and of the japonicarice (cv. Sasanishiki), which had been harvested 30 and 60 dafter anthesis. The inhibition of germination caused by dehuskingjaponica rice was overcome by 0.5–5% ethanol, with maximumgermination (frequently 100%) achieved at 3–5% (30 d afteranthesis) or 1–4.5% (60 d after anthesis) ethanol. Furtherincreases in the ethanol concentration reduced germination.The germination of dehusked indica rice was slightly inhibitedat 0.5 and 1% ethanol, whilst the promotion of germination by2% ethanol increased as the seeds matured. At all harvests germinationwas greatest at 3% ethanol, and at 5–6% ethanol germinationfell to 0%. Inhibition, no effect, or minimal stimulation ofthe germination of intact seeds of both japonica and indicarice by ethanol was observed at the concentrations examined.The absence of oxygen stimulated germination of dehusked japonicarice, but this germination was inhibited by ethanol. In contrastethanol had little or no effect on the failure of dehusked indicaseeds to germinate in anaerobic conditions. Thus ethanol treatmentmay help break the strong dormancy of dehusked seeds of indicaand japonica rice. The possible role of ethanol in stimulatinggermination in rice is discussed. Rice; Oryza sativa L.; seed germination; dehusking treatment; ethanol; indica; japonica; oxygen; dormancy; germination inhibition; seed formation     

THREE-DIMENSIONAL STRUCTURE OF EXTRACELLULAR MATRIX REVERSIBLY REGULATES MORPHOLOGY,PROLIFERATION AND COLLAGEN METABOLISM OF PERISINUSOIDAL STELLATE CELLS (VITAMIN A-STORING CELLS)

The three-dimensional structure of the extracellular substratum was found to regulate reversibly the morphology, proliferation and collagen synthesis of perisinusoidal stellate cells (lipocytes, i.e. fat-storing ‘Ito’ cells). On non-coated polystyrene and type I collagen-coated culture dishes, the cells spread well and extended their cellular processes. On the surface of type I collagen gels, the cells gathered and formed a mesh-like structure. However, in type I collagen gel where the cells were surrounded by type I collagen three-dimensionally, the cells extended their fine cellular processes and resembled the star-shaped stellate cells seenin vivo. The cell proliferation was more prominent in culture dishes coated with type I collagen or in polystyrene culture dishes than on or in type I collagen gels. The collagen synthesis was affected in the same manner. These data indicate that the nature and the three-dimensional structure of the extracellular matrix (ECM) can regulate morphology, proliferation and functions of the perisinusoidal stellate cells. In order to examine the reversibility of these regulations, we liberated cultured cells with trypsin or with purified bacterial collagenase and re-seeded them onto or into each substratum. The cells changed their shape, rate of proliferation and collagen synthesis according to each new substratum. These results indicate that the three-dimensional structure of ECM reversibly regulates the morphology, proliferation rate and functions of the perisinusoidal stellate cells.     

Biological activity of phyllosinol, a phytotoxic compound isolated from a culture filtrate of Phyllosticta sp.

1. Phyllosinol is a phytotoxic metabolite of Phyllosticta sp. Thissubstance at 100 µg/ml produced dark grey necrotic lesionson the leaf of red clover. Sensitivities of various plant speciesto phyllosinol differed both quantitatively and qualitatively.
2. Phyllosinol reduced root growth in rice seedlings by 60% at10^–4 M, whereas stimulation of root elongation occurredat a concentration range from 10^–9 to 10^–5 M.
3. Phyllosinolat 2.5x10^–4M promoted adventitious root formationin epicotylsof Azukia cuttings by about 100%. Promotion waspartly reducedby simultaneous application of cysteine.
4. IAA-induced elongationof isolated Avena coleoptile sectionswas inhibited by phyllosinolat a concentration range from 10^–5to 10^–3M.
5. Sulfhydrylcompounds, i.e. cysteine and glutathione relievedinhibitioncaused by phyllosinol in IAA-induced elongation ofAvena coleoptilesections.
6. GA₃-induced elongation of wheat leaf sections wasslightly inhibitedby phyllosinol at 10^–4M.
7. Phyllosinolalso has antibiotic activity. Among the organismstested, Phycomycetesand Gram-negative bacteria appeared mostsusceptible to phyllosinol.

(Received April 21, 1970; )     

Growth Response to Hormones by a New Rat Ovary Cell Line

SEVERAL endocrine cell lines established in recent years show a functional response to hormones in vitro¹ but, except for one mammary cell line², none of them exhibits the normal hormone requirement for growth in vivo. We have now isolated a rat ovarian cell line whose growth in vitro is markedly stimulated by bovine luteinizing hormone (LH-NIH-B7), a pituitary gonadotrophin and by dexamethasone, a synthetic glucocorticoid. This cell line provides the first permanent in vitro system for studying the growth stimulation of gonadal cells by hormones.