欧洲小叶椴的胚培养与快速繁殖

1植物名称欧洲小叶椴（Tilia cordata Mill．）。 2材料类别成熟胚。 3培养条件（1）／3动培养基：MS＋6-BA2．0mg·L^-1（单位下同）＋NAA0．1;（2）增殖培养基：MS＋6．BA1．0＋NAA0．1;（3）壮苗生根培养基：1／2MS＋IBA1．0＋NAA0．3。以上培养基均加入0．55％琼脂和20g·L^-1。蔗精,pH5．8-6．0。培养温度为（25±2）℃;光照培养时间12h·d^-1,光照强度40．50μmol·m^-2·s^-1。     

Anatomical study of zygotic and somatic embryos of Tilia cordata

A comparative anatomical study was carried out on zygotic and somatic embryos of Tilia cordata Mill. to evaluate the effect of growth conditions on their development. Zygotic embryos (heart-shaped, torpedo, cotyledonary), collected during two autumn periods, were examined to investigate the effect of growing season on embryo development. In comparison, the influence of growth conditions on the development of somatic embryos in vitro was also studied. Treatment with abscisic acid (ABA) and polyethylene glycol-4000 induced the development of somatic cotyledonary embryos similar to zygotic embryos with respect to morphology and anatomy, as illustrated by the differentiation of the apical meristems and of the procambium. The pattern of accumulation of starch and protein was also similar in these embryos. Somatic cotyledonary embryos that developed spontaneously without ABA showed defective accumulation of storage material and a general failure to form the shoot apical meristem, leading to very low germination rates. Vacuolar phenolic deposits were observed along the procambium of both zygotic and somatic embryos regardless of the maturation stage. Tracheid formation was observed only in somatic embryos formed without ABA in the medium and in precociously germinated somatic embryos. Phenolic vacuolar inclusions were frequently observed in epidermal cells of these embryos. This revised version was published online in June 2006 with corrections to the Cover Date.     

The identity of Rumicicarpus ramosissimus Chiov. (Tiliaceae)

Rumicicarpus ramosissimus Chiov. is found to be conspecific with Triumfetta trigona Sprague & Hutch. The latter name is to be used for the combined taxon, which belongs to the T. flavescens group of sect. Triumfetta. The distribution of T. trigona is mapped.     

In vitro propagation of oak (Quereus robur L.) and linden (Tilia cordata Mill.)

Rapid multiplication of axillary shoots of oak and linden has been achieved on broad-leaved tree medium (BTM) and woody plant medium (WPM) containing low level of cytokinin (BAP 0.2–1.0 mg l^-1). High rooting percentages (80–95%) were obtained on low salt, low sucrose media, containing low level of auxins. Rooted plants were transplanted into pots containing a mixture of peat and perlite. Most of the plants (90–95%) survived the transfer. After the hardening off period the new plants were planted in the field.     

Localization of cytokinins in somatic and zygotic embryos of Tilia cordata using immunocytochemistry

Immunofluorescent and immunogold labeling was used to study the localization of cytokinins in developing somatic and zygotic embryos of Tilia cordata Miller. Broad-specificity polyclonal antibodies active against dihydrozeatin riboside (DHZR)/zeatin riboside (ZR)-type and isopentenyladenosine (iPR)-type cytokinins were used for immunolabeling. Immunofluorescent microscopy showed that these cytokinins were concentrated in highly cytoplasmic cells showing meristematic character. Cotyledon initials and primary meristems of heart-stage somatic embryos, as well as heart-stage zygotic embryos, were labeled. During elongation of embryos, cytokinin immunoreactive material was concentrated to areas having meristematic character. Root apex, shoot meristem, and cotyledon cells of somatic and zygotic cotyledonary embryos, as well as epidermal and subepidermal cell layers of the hypocotyl, showed the strongest immunoreaction. The nucleoli, especially, had a very strong signal. Results at the ultrastructural level with gold-conjugated protein A supported these conclusions. Gold particles were distributed in the nuclei, especially in the nucleoli and throughout the ground cytoplasm. They were occasionally associated with plastids and mitochondria, but seldom with other organelles.     

Polyamine levels in buds and twigs of Tilia cordata from dormancy onset to bud break

The fluctuations of free and bound polyamines (PAs) were studied in vegetative buds and underlying twigs of linden (Tilia cordata L.) from August to May, to assess the connection between PA levels and seasonal cycles of growth and dormancy. Outer and inner bud scales and shoot tips (short shoot tips with leaf initials in contiguous short internodes) were analyzed separately, as were phloem with cortex and xylem with pith tissue from twigs. Seasonal variations in PA levels were present in buds and twigs during the research period. The most abundant PA in buds and twigs in free and bound forms was spermidine followed by putrescine. PA amounts were low in buds and twigs in autumn. In twig tissues, free PAs were predominant whereas in bud scales, bound PAs accumulated over free PAs in autumn, first in inner scales and later in outer scales as well. PA levels did not increase dramatically during the onset of dormancy in autumn but lower temperatures and probable cold hardening correlated positively with bound PAs in bud scales. In shoot tips with leaf initials, and contiguous short internodes, free putrescine and spermidine levels rose simultaneously with bud burst and new growth, while bound PAs diminished quite radically from temporary bud scales and from growing shoot tips.     

Wood specific gravity and anatomy in Heliocarpus appendiculatus (Tiliaceae)

Specific gravity exhibits extremely large radial increases with distance from the pith in Heliocarpus appendiculatus Turcz. (Tiliaceae), a pioneer of neotropical wet forests. To determine some of the wood anatomical changes associated with this increase, wood samples taken at breast height from three trees were divided into 1.0-cm-long segments from pith to bark. Measurements were made of fiber wall thickness, fiber lumen diameter, and percentages of fibers, axial parenchyma, ray parenchyma, and vessels on sections prepared from each segment. The extreme radial increases in specific gravity were associated with increases in fiber wall thickness, decreases in fiber diameter, decreases in fiber lumen diameter, and changes in the relative proportions of fibers and parenchyma. The increase in percent fiber concomitant with a decrease in axial parenchyma was the most important contributor to the increase in specific gravity in this species. The best predictor of specific gravity was percent fibers (r = 0.91, 0.92, 0.94) or percent axial parenchyma (r = -0.92, -0.91, -0.95), two variables that were highly intercorrelated (r = -0.95, -0.98, -0.99).     

Temperature and water content effects on the viscoelastic behavior of<Emphasis Type="Italic"> Tilia americana</Emphasis> (Tiliaceae) sapwood

The effects of temperature and water content on the viscoelasticity of living and dehydrated Tilia americana sapwood were examined using transient creep (time- and load-dependent deformation) tests under sustained bending loads. Creep tests were performed at 21.1°C and –20.5°C to determine the magnitudes and types of strains in living and dehydrated samples. Temperature had no effect on the creep rate of living sapwood. However, the creep rate of dehydrated samples at –20.5°C was significantly faster than that at 21.1°C. Regardless of temperature, sapwood had a faster creep rate than dehydrated samples. With small bending loads, the residual strains in sapwood were larger at 21.1°C compared to –20.5°C. Temperature did not significantly affect the residual strains in dehydrated samples. For small bending loads, frozen sapwood recovered all residual creep strains when thawed. With larger loads, residual and plastic (permanent) strains increased. We speculate that ice formation in cell lumens partially dehydrates (and thus stiffens and strengthens) cell wall materials and prevents cell wall buckling and elastic restoration after unloading. However, when thawed, sapwood can elastically restore its original configuration, provided it is not excessively bent (by ice or snow accumulations) when frozen.     

Chloroplast DNA Variation Shed Light on the History of Lime Tree (Tilia cordata s. l.) in the Eastern Part of the Range

Russian Journal of Genetics - The Quaternary history of European nemoral forests in the east of their distribution may differ significantly from the typical dynamics of populations and ranges of...     

New Mexican species of Byttneria (Sterculiaceae), Bakeridesia (Malvaceae), and Triumfetta (Tiliaceae)

Recent collections have brought to light the following new species from the states of Tabasco and Oaxaca, Mexico:Byttneria fluvialis (Sterculiaceae) from Tabasco;Bakeridesia amoena (Malvaceae), andTriumfetta calzadae (Tiliaceae) from Oaxaca.     

A new species of Apeiba (Tiliaceae) from the Venezuelan-Brazilian border

Apeiba uittienii is described as new and illustrated. This species is morphologically somewhat intermediate between the two sections formerly recognized in this genus. A revised key to the species ofApeiba (now eight in number) is provided.     

Hybridization among cryptic species of the cellar fungus Coniophora puteana (Basidiomycota)

In this study we have analysed the genetic variation and phylogeography in a global sample of the cellar fungus Coniophora puteana, which is an important destroyer of wooden materials indoor. Multilocus genealogies of three DNA regions (beta tubulin, nrDNA ITS and translation elongation factor 1alpha) revealed the occurrence of three cryptic species (PS1-3) in the morphotaxon C. puteana. One of the lineages (PS3) is apparently restricted to North America while the other two (PS1-2) have wider distributions on multiple continents. Interspecific hybridization has happened between two of the lineages (PS1 and PS3) in North America. In three dikaryotic isolates, two highly divergent beta tubulin alleles coexisted, one derived from PS1 and one from PS3. Furthermore, one isolate included a recombinant ITS sequence, where ITS1 resembled the ITS1 version of PS3 while ITS2 was identical to a frequent PS1 ITS2 version. This pattern must be due to hybridization succeeded by intralocus recombination in ITS. The results further indicated that introgression has happened between subgroups appearing in PS1. We hypothesize that the observed reticulate evolution is due to previous allopatric separation followed by more recent reoccurrence in sympatry, where barriers to gene flow have not yet evolved. A complex phylogeographical structure is observed in the morphotaxon C. puteana caused by (i) cryptic speciation; (ii) the interplay between natural migration and distribution patterns and probably more recent human mediated dispersal events; and (iii) hybridization and introgression.     

Influence of root temperature on growth of Pinus sylvestris,Fagus sylvatica,Tilia cordata and Quercus robur

One-year-old tree seedlings were incubated in a greenhouse from April to July, under natural daylight conditions, with their root systems at constant temperatures of 5, 10, 15, 20, 25, 30 and 35 °C and with the above ground parts kept at a constant air temperature of 18–20 °C. The course of height growth, total mass increment, root, shoot and leaf weight as well as leaf areas were measured. The results indicate that clear differences exist in the optimal root zone temperatures for various growth parameters in different tree species. Pinus sylvestris had a maximal height increment at about 5–10 °C and maximal total mass increment at 15 °C root temperature. In contrast, the optimum for Quercus robur was at 25 °C. Tilia cordata and Fagus sylvatica had their optima for most growth parameters at 20 °C. The root temperature apparently indirectly influenced photosynthesis (dry weight accumulation) and respiration loss. From the observed symptoms and indications in the literature it seems probable that a change in hormone levels is involved as the main factor in the described effects. Variation of root temperature had only an insignificant effect on bud burst and the time at which the shoots sprouted. Apparently species of northern origin seem to have lower root temperature optima than those of more southern origin. This is to be verified by investigation of other tree species.     

Hybridization studies inSilene subgen.Petrocoptis (Caryophyllaceae)

Silene subgenusPetrocoptis comprises sexual diploid taxa and it is restricted to calcareous cliffs in the Iberian peninsula. Artificial crosses involvingSilene pyrenaica (Bergeret)Mayol etRosselló (≡Petrocoptis pyrenaica (Bergeret)Walp.),Silene laxipruinosa Mayol etRosselló andS. montserratii subsp.crassifolia (Rouy)Mayol etRosselló (≡P. crassifolia Rouy) were attempted to assess the extent of barriers developed within the subgenusPetrocoptis. Usually, intraspecific crosses involving allopatric populations were successful, suggesting that geographically isolated populations are not genetically isolated. Cross-compatibility was noted among the polymorphicS. pyrenaica, which developed fertile F₁ hybrids. All other interspecific crosses failed due to cross- or seed-incompatibility. Crossing results agree with available evidence supporting the merging of segregates ofS. pyrenaica within a single taxon.     

Hybridization betweenPsidium guajava andP. guineense (Myrtaceae)

Psidium guajava andP. guineense are characterized, illustrated and contrasted. Population samples ofPsidium guajava andP. guineense and apparent hybrids were collected from four localities: two in Mexico, one in Honduras, and one in Argentina. Seven morphological characters, one micromorphological (stomatal density), and one chemical character (presence or absence of myricetin) that distinguish the parent species were found to be useful in the evaluation of specimens and population variation. Results were consistent with the hypothesis that hybridization occurs between these species, but in three localities it may not go beyond Fl ’s. Back crossing between hybrids andP. guajava was not detected at any locality.     

Patterns of pollen removal and deposition in tristylous Pontederia cordata L. (Pontederiaceae)

In tristylous Pontederia cordata (Pontederiaceae), conspicuous differences in the size of pollen grains and discrete variation in the length of reproductive organs provide a suitable experimental system for the study of fine-scale pollination events. At a population of P. cordata at Pothole Lake, Ontario, the majority of flowers are visited by bumble bees which remove on average 45% of the pollen during single visits to previously unvisited flowers. The amount and proportion of pollen removed are significantly different among floral morphs and stamen levels. Deposition of the three pollen types on the bodies of Bombus spp., Apis mellifera and Melissodes apicata is non-random: large- and medium-size pollen tends to remain in greatest concentrations where it is initially deposited, whereas small-size pollen is displaced from the proboscis to more posterior body parts, probably as a result of grooming activities. Stigmatic pollen loads of individual flowers following single bumble bee visits indicate that the mid-styled morph captures the largest total pollen load, and the short-styled morph the smallest. The largest proportion of compatible pollen grains is deposited on stigmas of the long-styled morph. Pollen load data from "single visit" flowers is in general agreement with previously published population surveys involving multiply-visited flowers.     

The function and adaptive significance of tristyly in Pontederia cordata L. (Pontederiaceae)

Darwin proposed that the adaptive significance of tristyly is to promote insect-mediated pollination among floral morphs with anthers and stigmas at equivalent levels (legitimate pollination). Strong pollen trimorphism in Pontederia cordata enables an evaluation of this hypothesis. In an investigation of pollen flow patterns in different parts of the North American range of the species, legitimate pollination of some morphs was observed in all but one population investigated. However at only one site (Ft. McCoy, Florida) was Darwin's hypothesis confirmed for the three floral morphs. The long-styled morph most frequently exhibited legitimate pollination, the mid-styled morph was intermediate, and the short-styled morph rarely experienced legitimate pollination.
Temporal variation in pollen flow was observed during the flowering season of P. cordata . Total stigmatic pollen loads decreased as the density of inflorescences and activity of pollinators declined. In spite of variable total loads the legitimate component was relatively constant in the long- and mid-styled forms, while in the short-styled form this measure increased as the season progressed. Little variation was detected between stigmatic samples collected at different times during a single day.
Legitimate pollination may be inhibited by local foraging of pollinators and the spatial segregation of floral morphs. At Paugh Lake (Ontario) over 75% of all bumble bee flights are among the five nearest neighbours of a particular inflorescence. In the same population, there is a probability of over 70% that the nearest three neighbours of an inflorescence are of the same floral morph. Despite these influences, legitimate pollination occurs in P. cordata , suggesting substantial pollen carryover.     

Fluorescence in Situ Hybridization (FISH): DNA Probe Production and Hybridization Criteria

We describe methods for the production of fluorescence in situ hybridization (FISH) probes and the utilization of these probes for the detection of complementary DNA sequences with accuracy and sensitivity for application in both basic research and clinical diagnosis. Due to the frequent use of FISH in many laboratories, it is important to apply the most convenient and reproducible approach. This review describes some of the most recent techniques, and includes versatile, effective and simple methods of probe production and fluorescence in situ hybridization. We also describe methods for the production of region-specific and chromosome-specific DNA probes and hybridization techniques for the visualization of these probes.