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PubMed2000

Isolation, characterization, and inheritance of microsatellite loci in alpine larch and western larch.

Microsatellite loci or simple sequence repeat loci (SSRs) were isolated in alpine larch (Larix lyallii Parl.) and western larch (Larix occidentalis Nutt.). In total, 14 SSR loci were characterized; two [(TCT)4, A7] came from published Larix DNA sequence data, one (CA)17 was obtained from a partial non-enriched alpine larch total genomic DNA library, and the remaining 11 loci were obtained from larch genomic DNAs enriched for (CA)n repeats. The SSR regions in these clones could be divided into three categories: perfect repeat sequences without interruption, imperfect repeat sequences with interruption(s), and compound repeat sequences with adjacent tandem simple dinucleotides. Eight of the 14 loci analyzed were found to be polymorphic and useful markers after silver-staining polyacrylamide gel electrophoresis. In addition, several SSR primers developed for alpine larch were able to successfully amplify polymorphic loci in its related species, western larch, and among other closely related taxa within the Larix genus. The inheritance of microsatellite loci was verified by analysis of haploid megagametophyte and diploid embryo tissues of progeny obtained from controlled crosses between western larch and alpine larch. All microsatellite loci analyzed had alleles that segregated according to expected Mendelian frequencies. Two species-specific markers (UAKLly10a and UAKLla1) allow easy and rapid identification of specific genetic entry of alpine larch and western larch at any stage in the sporophyte phase of the life cycle. Therefore, these markers are efficient in identifying the parental species and to validate controlled crosses between these two closely related species. These results are important in tree improvement programs of alpine larch and western larch aimed at producing genetically improved hybrid stock for reforestation in Western Canada and U.S.A.

Cloning, Molecular↗
PubMed2000

Effects of experimental conditions on mycorrhizal relationships between Pinus sylvestris and Lactarius deliciosus and unprecedented fruit-body formation of the Saffron milk cap under controlled soilless conditions.

The mycorrhizal relationships between pines and two edible species of Lactarius sect. Dapetes were investigated by optimizing the experimental conditions of mycelial growth and of mycorrhizal colonization of pine seedlings. In vitro mycelial growth of Lactarius deliciosus and L. sanguifluus was improved on a buffered medium containing glucose, amino acids, and vitamins. Two methods of mycorrhization of pines with Lactarius deliciosus were tested. The mycorrhizal colonization was rapid and intense under non-aseptic conditions with a low nutrient supply and without exogenous glucose. A positive influence of mycorrhizal colonization on Pinus sylvestris growth was subsequently observed. Under axenic conditions and with a high nutrient supply, mycorrhization was stimulated at 10 g/L of exogenous glucose, irrespective of the phosphorus concentration. At high phosphorus level (1 mM) and 0.1, 1.0, or 10.0 g/L glucose, growth of Pinus sylvestris was reduced by inoculation. Stability and development of Pinus spp./Lactarius deliciosus symbioses were assayed in a climatic chamber using containers filled with a synthetic substrate. Over a 2-year culture period, the root systems of the pine seedlings were heavily colonized by Lactarius deliciosus. One year following inoculation, Lactarius deliciosus fruit-body primordia appeared associated with Pinus sylvestris seedlings. Six months later, two mature basidiomata were obtained. This is the first report of soilless fruit-body formation of this edible mushroom.

Basidiomycota↗
PubMed1997

Effects of shade and root confinement on the expression of plagiotropic growth in juvenile-origin Douglas-fir rooted cuttings.

The purpose of this experiment was to determine why juvenile-origin Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) rooted cuttings, which remain plagiotropic (branchlike) when grown in containers in shaded glasshouses, become orthotropic (vertical) after they are transplanted to an outdoor environment. Plagiotropic rooted cuttings (mean angle from vertical = 45-50 degrees) from three full-sib families were transplanted into an outdoor nursery and subjected to four treatments consisting of a factorial of (1) shaded or unshaded and (2) bareroot or confined roots. After two growing seasons, treatments had significantly affected plant size and biomass in the order unshaded-bareroot > shaded-bareroot > unshaded-confined > shaded-confined, but plants in all treatments had become nearly orthotropic. It is concluded that neither shading nor root confinement is, but other glasshouse environmental conditions are, responsible for the persistence of plagiotropic growth.

Biomass↗
PubMed2003

Photosynthesis of overwintering evergreen plants.

In this review we focus on photosynthetic behavior of overwintering evergreens with an emphasis on both the acclimative responses of photosynthesis to cold and the winter behavior of photosynthesis in conifers. Photosynthetic acclimation is discussed in terms of the requirement for a balance between the energy absorbed through largely temperature-insensitive photochemical processes and the energy used for temperature-sensitive biochemical processes and growth. Cold acclimation transforms the xanthophyll-mediated nonphotochemical antenna quenching of absorbed light from a short-term dynamic response to a long-term sustained quenching for the whole winter period. This acclimative response helps protect the evergreen foliage from photooxidative damage during the winter when photosynthesis is restricted or prevented by low temperatures. Although the molecular mechanisms behind the sustained winter excitation quenching are largely unknown, it does involve major alterations in the organization and composition of the photosystem II antenna. In addition, photosystem I may play an important role in overwintering evergreens not only by quenching absorbed light photochemically via its support of cyclic electron transport at low temperatures, but also by nonphotochemical quenching of absorbed light irrespective of temperature. The possible role of photosystem II reaction centers in nonphotochemical quenching of absorbed energy in overwintering evergreens is also discussed. Processes like chlororespiration and cyclic electron transport may also be important for maintaining the functional integrity of the photosynthetic apparatus of overwintering evergreens both during periods of thawing in winter and during recovery from winter stress in spring. We suggest that the photosynthetic acclimation responses of overwintering evergreens represent specific evolutionary adaptations for plant species that invest in the long-term maintenance of leaf structure in cold climatic zones as exemplified by the boreal forests of the Northern Hemisphere.

Acclimatization↗
PubMed2004

Rhamnogalacturonan II: structure and function of a borate cross-linked cell wall pectic polysaccharide.

Rhamnogalacturonan II (RG-II) is a structurally complex pectic polysaccharide that was first identified in 1978 as a quantitatively minor component of suspension-cultured sycamore cell walls. Subsequent studies have shown that RG-II is present in the primary walls of angiosperms, gymnosperms, lycophytes, and pteridophytes and that its glycosyl sequence is conserved in all vascular plants examined to date. This is remarkable because RG-II is composed of at least 12 different glycosyl residues linked together by more than 20 different glycosidic linkages. However, only a few of the genes and proteins required for RG-II biosynthesis have been identified. The demonstration that RG-II exists in primary walls as a dimer that is covalently cross-linked by a borate diester was a major advance in our understanding of the structure and function of this pectic polysaccharide. Dimer formation results in the cross-linking of the two homogalacturonan chains upon which the RG-II molecules are constructed and is required for the formation of a three-dimensional pectic network in muro. This network contributes to the mechanical properties of the primary wall and is required for normal plant growth and development. Indeed, changes in wall properties that result from decreased borate cross-linking of pectin may lead to many of the symptoms associated with boron deficiency in plants.

Cycadopsida↗
PubMed2001

Cupressaceae pollinosis: identification, purification and cloning of relevant allergens.

Allergy to Cupressaceae pollen is a worldwide pollinosis caused by several species. Pollen extracts prepared from allergenic species belonging to this family are characterised by low protein and high carbohydrate content. The allergenic components represented in the pollen extracts from different species of the Cupressaceae family show high levels of cross-reactivity when probed with human IgE from allergic subjects and share a number of common epitopes also identified by polyclonal rabbit antisera and monoclonal antibodies. A close relationship has also been described with the Taxodiaceae and Podocarpaceae families. Although both proteic and carbohydrate epitopes appear to be involved in IgE recognition and allergenic cross-reactivity, a large portion of the IgE reactivity of Cupressaceae-allergic patients seems to be associated with sugar moieties present on the relevant allergenic molecules. From this point of view, Cupressaceae/Taxodiaceae allergens constitute a particularly useful model to study IgE cross-reactivity, as they have been shown to display different levels of homology. Moreover, the availability of the purified allergens, together with their recombinant counterparts, may shed light on the actual role played by carbohydrate in allergic sensitisation, IgE recognition and allergenic cross-reactivity.

Allergens↗
PubMed2003

The PCR-based detection of Leishmania major infections in Meriones libycus (Rodentia: Muridae) from southern Iran.

Cutaneous leishmaniasis (CL) is an increasing public-health problem in the countries of the Mediterranean region. In southern Iran, for example, the incidence of CL has more than doubled over the last decade. As part of an investigation of this worrying trend, the leishmanial infections of the wild rodents that may act as reservoir hosts were investigated in the Iranian province of Fars. Overall, 82 rodents, 56 of them Libyan jirds (Meriones libycus), were collected, in live traps, from a new, hypo-endemic focus of zoonotic CL. When Giemsa-stained smears of ear tissue or any lesions on the rodents were prepared and checked under a light microscope, amastigotes were found in the smears from six of the rodents, all M. libycus. None of the other rodents investigated was found to be infected. The infected rodents were encountered in two of the three areas studied. In PCR-based assays, the amastigotes seen in the ear smears and those from two recent human cases of CL from the focus were all found to be identical to a reference strain of L. major. It therefore seems that M. libycus is the main active reservoir host for ZCL in the focus.

Animals↗
PubMed2001

Richness of Colchic vegetation: comparison between refugia of south-western and East Asia.

BACKGROUND: The Colchis is one of the species-rich refugia and a centre of biological diversity in western Eurasia. We analysed patterns of richness, endemism and invasions in relation to taxonomy (family membership), life form, certain habitats in the Colchis, and compared them to patterns found for Japan. RESULTS: We found that in the Colchis perennials are significantly over-represented in endemic species, and that they typically occur on limestone soils and in alpine tall herbaceous vegetation. The Asteraceae produce significantly large number of both endemic and alien species, whereas the Poaceae are over-represented in alien species but under-represented in endemics. Likewise, the Apiaceae are over-represented in endemics, whereas the Euphorbiaceae are over-represented in alien species. Similar patterns have been found in Yakushima, Japan. The Morisita-Horn index of similarity between these two sites was 0.83 (based on family size). Although the flora of Adjara comprised of fewer families than the flora of Yakushima, the largest families are richer in species in the flora of Adjara than in the flora of Yakushima. CONCLUSIONS: Floristic analysis of refugia of western Eurasia and their comparison with geographically distant areas can provide useful data for plant ecological and evolutionary studies. Potentially, such studies can produce testable hypotheses on plant migrations and on their historical geography. For example, the data presented in this study indicate that more severe conditions in the Pleistocene and geographical isolation of the Colchis may be responsible for the higher relative importance of adaptive radiation in the shaping of its modern flora.

Biodiversity↗
PubMed2003

Expressed sequence tag analysis in Cycas, the most primitive living seed plant.

BACKGROUND: Cycads are ancient seed plants (living fossils) with origins in the Paleozoic. Cycads are sometimes considered a 'missing link' as they exhibit characteristics intermediate between vascular non-seed plants and the more derived seed plants. Cycads have also been implicated as the source of 'Guam's dementia', possibly due to the production of S(+)-beta-methyl-alpha, beta-diaminopropionic acid (BMAA), which is an agonist of animal glutamate receptors. RESULTS: A total of 4,200 expressed sequence tags (ESTs) were created from Cycas rumphii and clustered into 2,458 contigs, of which 1,764 had low-stringency BLAST similarity to other plant genes. Among those cycad contigs with similarity to plant genes, 1,718 cycad 'hits' are to angiosperms, 1,310 match genes in gymnosperms and 734 match lower (non-seed) plants. Forty-six contigs were found that matched only genes in lower plants and gymnosperms. Upon obtaining the complete sequence from the clones of 37/46 contigs, 14 still matched only gymnosperms. Among those cycad contigs common to higher plants, ESTs were discovered that correspond to those involved in development and signaling in present-day flowering plants. We purified a cycad EST for a glutamate receptor (GLR)-like gene, as well as ESTs potentially involved in the synthesis of the GLR agonist BMAA. CONCLUSIONS: Analysis of cycad ESTs has uncovered conserved and potentially novel genes. Furthermore, the presence of a glutamate receptor agonist, as well as a glutamate receptor-like gene in cycads, supports the hypothesis that such neuroactive plant products are not merely herbivore deterrents but may also serve a role in plant signaling.

Amino Acids, Diamino↗
PubMed2004

A new structural element containing glycine-rich proteins and rhamnogalacturonan I in the protoxylem of seed plants.

The water pipes of elongating plant organs are the result of programmed cell death and are formed by the walls of dead and empty protoxylem elements. These protoxylem elements are passively elongated many times by the surrounding tissue before they are replaced and collapse. Well-known adaptations for this unique task include the characteristic secondary wall thickenings, forming rings and helices. A new, clearly distinct structural element containing glycine-rich proteins is now visualized for the first time, using confocal laser scanning microscopy in the mature protoxylem of elongating organs of seed plants. This structural element is arranged along the longitudinal axis of the protoxylem elements. It interconnects the secondary wall thickenings within and between protoxylem elements, as well as the protoxylem with other cell types such as xylem parenchyma cells and metaxylem elements. The structural element is stable against detergent extractions, proteinase, pectinase and cellulase hydrolysis, and is closely associated with rhamnogalacturonan-I, a pectic polysaccharide. The results clearly demonstrate that the cell wall of protoxylem cells is a highly dynamic and complex structure. The typical polysaccharide-rich primary wall of living and elongating plant cells is progressively modified and finally replaced by a protein-rich wall in the dead and passively stretched protoxylem elements. These glycine-rich walls originated early in the evolution of the seed plants as confirmed by the analysis of genomic information.

Amino Acid Sequence↗
PubMed1999

Antimicrobial activity and metalloprotease inhibition of hinokitiol-related compounds, the constituents of Thujopsis dolabrata S. and Z. hondai MAK.

Gamma-thujaplicin, beta-dolabrin and hinokitiol(beta-thujaplicin), hinokitiol-related compounds isolated from the wood of Thujopsis dolabrata S. and Z. hondai MAK have antimicrobial activity. In particular, strong antibacterial activity of hinokitiol and beta-dolabrin on Staphylococcus epidermidis IFO-12993 was found, with a minimum inhibitory concentration (MIC) of 0.2 microg/ml. This activity was higher than that of gentamicin, used as a positive control, and so the strong antibacterial activity of both compounds on this bacterium is of considerable interest. Of the three compounds, gamma-thujaplicin showed the strongest antifungal activity and its MIC was found to be around 1.5 microg/ml. The three compounds also inhibited metalloproteases. The inhibitory activity of hinokitiol on carboxypeptidase A was especially strong, its 50%-inhibitory concentration (IC50) being 2.76x10(-6) M. Considering that metalloproteases are involved in inflammation, the strong inhibitory activity of hinokitiol could be important. On the other hand, hinokitiol-acetate did not show any antimicrobial activity and metalloprotease inhibition, suggesting that at least part of the activity is due to metal chelation between the carbonyl group at C-1 and the hydroxyl group at C-2 in the tropolone skeleton.

Anti-Bacterial Agents↗
PubMed2000

Isolation of the antiulcer compound in essential oil from the leaves of Cryptomeria japonica.

Essential oil from the leaves of Tateyamasugi (Cryptomeria japonica) exhibited strong inhibitory activity on ulceration induced by HCl/ethanol, HCl/aspirin, water-immersion stress and pylorus-ligation. We separated the antiulcer compounds from cedar essential oil by use of distillation and chromatography. As a result, terpinen-4-ol, a monoterpene, and elemol, a sesquiterpene, were isolated as active compounds. The antiulcer activity of the former was more potent than that of the latter. Terpinen-4-ol was a mixture of optical isomers and each possessed potent antiulcer activity. Secretion of gastric juice and output of acid and pepsin activity were lowered by terpinen-4-ol. These results suggest that terpinen-4-ol isolated from cedar essential oil could be a valuable antiulcer agent.

Acetaldehyde↗
PubMed2000

Phytogrowth-Inhibitory activities of beta-dolabrin and gamma-thujaplicin, hinokitiol-related compounds and constituents of Thujopsis dolabrata Sieb. et Zucc. var hondai Makino.

Beta-dolabrin and gamma-thujaplicin isolated from Thujopsis dolabrata Sieb. et Zucc. var hondai Makino, like hinokitiol, showed strong phytogrowth-inhibitory activities, and their growth-inhibitory activities were as high as that of sodium 2,4-dichlorophenoxyacetate used as a positive control. In particular, the phytogrowth-inhibitory activity of gamma-thujaplicin was strong and it completely inhibited the germination of this seed of Brassica campestris L. subsp. rapa Hook f. et Anders at the concentration of 30 ppm. Both compounds exhibited inhibitory activities on B. campestris L. subsp. rapa Hook f. et Anders and Sesamum indicum Linne, even at the low concentration of 10 ppm. At 7 d after treatment with beta-dolabrin and gamma-thujaplicin, the amount of chlorophyll in the cotyledons of B. campestris L. subsp. rapa Hook f. et Anders treated with both compounds was greatly decreased as compared with the control. The findings indicate that the phytogrowth-inhibitory action might be a common biological activity of hinokitiol-related compounds, suggesting that at least a part of their phytogrowth-inhibitory actions seems to be related to a decrease in chlorophyll content.

Brassica↗
PubMed2000

Novel A-seco-rearranged lanostane triterpenoids from Abies sachalinensis.

From the needles of Abies sachalinensis, novel rearranged lanostane type triterpenes, 1-4, were isolated along with a known triterpene (5). The structures of the new compounds, 1-4, were elucidated to be 3,4-seco-8-(14-->13R)abeo-17,13-friedo-9beta-lanosta-4(28),7,14(30),22Z,24-pentaen-26,23-olide-3-oic acid, methyl 3,4-seco-8-(14-->13R)abeo-17,13-friedo-9beta-lanosta-4(28),7,14(30),22Z,24-penten-26,23-olide-3-oate, 3,4-seco-8(14-->13R)abeo-17,13-friedo-9beta-lanosta-4(28),7,14,22Z,24-pentaene-26,23-olide-3-oic acid and methyl 3,4-seco-8(14-->13R)abeo-17,13-friedo-9beta-lanosta-4(28),7,14,22Z,24-pentaene-26,23-olide-3-oate, respectively, by means of spectral experiments, especially two dimensional NMR spectroscopy, such as 1H-detected multiple quantum coherence (HMQC), 1H-detected heteronuclear multiple bond connectivity (HMBC) and 1H-1H-correlation spectroscopy (COSY) experiments. These new compounds have novel structures containing A-seco, rearranged spiro structure and a gamma-lactone conjugated with a diene. Some of these compounds showed potent antibacterial activity against gram positive bacteria.

Anti-Bacterial Agents↗
PubMed2002

Stilbene derivatives from two species of Gnetaceae.

Five new stilbene oligomers (gnemonols A, B and C, gnemonoside E and gnetal) were isolated together with 2b-hydroxyampelopsin F and gnetin E from Gnetum gnemon and G. gnemonoides. The structures of the compounds were elucidated on the basis of spectral evidence.

Benzofurans↗
PubMed1998

Enzymatic properties of a Ginkgo biloba endo-beta-N-acetylglucosaminidase and N-glycan structures of storage glycoproteins in the seeds.

An endo-beta-N-acetylglucosaminidase has been purified to homogeneity from mature seeds of Ginkgo biloba. The molecular mass of the endo-beta-N-acetylglucosaminidase, named Endo-GB, was estimated to be around 63 kDa by SDS-PAGE and around 62 kDa by Hiprep S-200 chromatography, respectively. The substrate specificity has been explored with regard to the pyridylaminated N-glycans. Several high mannose-type sugar chains bearing alpha-1,2-mannosyl residue(s), Man9-6GlcNAc2-PA, were the most favored substrates followed by Man5GlcNAc2-PA and a typical hybrid-type structure (GlcNAc1Man5Glc NAc2-PA) which does not bear an alpha-1,2-mannosyl residue. On the contrary, endo-GB could hardly hydrolyze the common core pentasaccharide of N-glycan (Man3GlcNAc2-PA) and the xylose-containing sugar chains (Man4-3Xyl1Glc NAc2-PA, Man3Fuc1Xyl1GlcNAc2-PA) being widely distributed in plant glycoproteins. Furthermore, we analyzed the structures of N-glycans conjugated to storage glycoproteins in the mature Ginkgo seeds to see the occurrence of endogenous substrates for Endo-GB. The structural analysis showed, however, only xylose-containing type N-glycans (Man3Fuc1Xyl1GlcNAc2 (95%) and Man3Xyl1 GlcNAc2 (5%)), which can not be substrate for Endo-GB, predominantly occur in the storage glycoproteins.

Carbohydrate Sequence↗
PubMed2000

Experimental sensitization with Japanese cedar pollen in dogs.

Japanese cedar pollinosis is a type I allergic disease mediated by immunoglobulin E (IgE) antibodies to Japanese cedar (Cryptomeria japonica) pollen antigen (CPAg). By using 22 dogs consisting of 20 dogs aged 3 months and 2 dogs aged 3 years, immunization was performed by subcutaneous injections of CPAg with aluminum hydroxide gel. Variable levels of CPAg-specific IgE antibody response were detected in 21 of the 22 immunized dogs two weeks after the second immunization. This study provided an experimental sensitization system with CPAg in dogs, which will be useful for further immunological studies on Japanese cedar pollinosis.

Animals↗
PubMed2001

Seasonal rhinitis in a cat sensitized to Japanese cedar (Cryptomeria japonica) pollen.

A cat showing seasonal allergic symptoms of rhinitis was examined for reactivities to Japanese cedar (Cryptomeria japonica, CJ) pollen allergen by intradermal skin test (IDST), Prausnitz-Kustner (P-K) test, and lymphocyte blastogenic response. In IDST for 26 common allergens. the cat showed a positive reaction to CJ pollen allergen. P-K test using CJ pollen allergen also showed a positive reaction, indicating the presence of serum IgE specific to CJ pollen. In the lymphocyte blastogenic response, the stimulation index in the presence of CJ pollen allergen was 2.4. These data suggested that the seasonal rhinitis observed in the cat was caused by the sensitization to CJ pollen allergen.

Animals↗