Socialization in spectacled parrotlets (Forpus conspicillatus): how juveniles compensate for the lack of siblings

  Socialization, that is, the process of the integration of an individual into the social system and the acquisition of basic social traits, has been studied in a variety of vertebrate species. Deprivation experiments have shown that the lack of early affiliative relationships has negative long-term effects on social behaviour and reproduction. Most studies emphasized the importance of the mother-infant relationship. However, it soon became evident that peer relationships to like-aged conspecifics, such as siblings and unrelated juveniles of nearly the same age, are also important for socialization. Whether individuals, deprived of one of these important social components, are able to compensate for this lack is not known. Here I tested the hypothesis that in spectacled parrotlets (Forpus conspicillatus), singletons (i.e., those that grew up without siblings) were able to compensate for this social lack by establishing and strengthening other relationships. In spectacled parrotlets socialization depends on the formation of long-lasting sibling groups. However, singletons without nest mates are quite common (about 6%), both in captivity and in nature. In this study I analysed the affiliative interactions with other group members of five singletons in comparison to eight siblings, from fledging to the 36th week after fledging, within a group of adult and juvenile conspecifics. In contrast to siblings, the singletons prolonged the relationships with their parents. Then they established a ”host sibling group” with other like-aged individuals. All singletons became well integrated into the group and established exclusive pair bonds. It is concluded that juvenile singletons of spectacled parrotlets are able to compensate for the lack of siblings by the development of alternative socialization tactics. Received: 12 December 1998 / Received in revised form: 17 May 1999 / Accepted: 7 June 1999     

Some properties of a pheromone allowing individual recognition, from the scats of an Australian lizard, Egernia striolata

  The Australian lizard, Egernia striolata, can distinguish its own scats from those of unfamiliar conspecific individuals. This appears to be unrelated to diet, because there is no difference in the response to scats from unfamiliar lizards fed on diets that are the same or different from the test lizard. The signal that induces the response is not a visual or tactile property of scat structure, because test lizards respond equally to crushed and intact scats. We suggest that a pheromone is secreted onto the scat as it is produced. Water solutions of scats did not contain signal components that allowed lizards to distinguish their own scats from others. However, solutions of scats in dichloromethane (DCM) retained unique characteristics, and test lizards responded more strongly to the solution from scats of an unfamiliar lizard that to the solution from their own scats. Further fractionation of the DCM solution in pentane and in methanol led to loss of the unique signals needed for individual recognition, but those were restored when the pentane and methanol fractions were recombined. We infer that these lizards can distinguish between scats of different individuals on the basis of signals they receive from a complex combination of chemicals. Received: 14 December 1998 / Received in revised form: 9 April 1999 / Accepted: 12 April 1999     

MS32-regulated timing of callose degradation during microsporogenesisin Arabidopsis is associated with the accumulation of stacked rough ER in tapetal cells

  In the male sterile32(ms32)mutant in Arabidopsis thaliana, pollen development is affected during meiosis of pollen mother cells (PMCs). In normal wild-type (WT) anthers, callose is deposited around PMCs before and during meiosis, and after meiosis the tetrads have a complete callose wall. In ms32, PMCs showed initial signs of some callose deposition before meiosis, but it was degraded soon after, as was part of the cellulosic wall around the PMCs. The early dissolution of callose in ms32 was associated with the occurrence of extensive stacks of rough ER (RER) in tapetal cells. The stacks of RER were also observed in the WT tapetum, but at a later stage, i.e., after the tetrads were formed and when callose is normally broken down for release of microspores. Based on these observations it is suggested that: (1) callose degradation around developing microspores is linked to the formation of RER in tapetal cells, which presumably synthesize and/or secrete callase into the anther locule, and (2) mutation in MS32 disrupts the timing of these events. Received: 27 April 1999 / Revision accepted: 21 June 1999     

New hyperbenthic species of Misophriopsis and Misophriella, first record of misophrioid copepods (Crustacea) from Antarctic waters

  Misophrioid copepods are reported for the first time from Antarctic waters. A species of each of the genera Misophriopsis and Misophriella is described from hyperbenthic layers of the continental shelf of the eastern Weddell Sea. The genus Misophria is also reported from these waters. Misophriopsis australis sp. nov. is the fifth representative of this widely distributed genus, previously reported from the North Atlantic, the China Sea, and both NW and NE quadrants of the Pacific Ocean. Misophriella schminkei sp. nov. is the second representative of the genus, known to date only from a single female caught in North Atlantic bathyal depths. Traces of subdivisions on antennary exopodal segments of the latter species suggest that the first segment may be a triple segment, while the second and terminal segments may be double segments. In addition, the mandibular exopod of this taxon indicates that a six-segmented ramus with setal formula 0, 1, 1, 1, 1, 2 may represent the ground pattern in Copepoda. The presence of parts of a cyclopinid copepod in the gut of M. schminkei sp. nov. indicates that this species may be carnivorous. Received: 12 November 1998 / Received in revised form: 26 July 1999 / Accepted: 27 July 1999     

The use of fish metabolic, pathological and parasitological indices in pollution monitoring

  An integrated biological effect monitoring concept has been tested in flounder (Platichthys flesus L.) from four locations with different anthropogenic impact in the German Bight. During 3 years of sampling, biomarkers at all levels of biological organisation from the molecular to the ecosystem level were applied and tested on 742 individual fish of body lengths between 18 and 25 cm. At the ecosystem level, the fish were taken as a habitat for the parasite assemblage. The hypothesis was that changes in the environment might lead to changes in the species diversity of parasites and in the infection intensity of single species, as well as between heteroxenic and monoxenic parasite species (H/M ratio). At the molecular level, activity of the CYP1A-dependent monooxygenase ethoxyresorufin O-deethylase (EROD) was used as a biomarker of exposure. At the subcellular level, the integrity of lysosomal membranes in hepatocytes was taken as an indicator of non-specific acute and chronic toxic effects. Both biomarkers are recommended by the ICES Advisory Committee on the Marine Environment for the application in biological effects monitoring programmes. In addition, neutral lipid content in the liver was used as a marker for pathologically induced fat accumulation. In the same individual fish, a new method for the measurement of macrophage aggregate activity in the liver was tested for its application and reliability in reflecting immunosuppression. Tests were accompanied by chemical analysis of standard organochlorine and heavy metal residues in flounder tissue. A total of 33 parasite species were found. As an indicator species, the mean abundance of Trichodina sp. reflected best the pollution gradient observed with highest infection intensity at the most polluted location. Species diversity was significantly higher in fish caught near the reference site and significantly lower in fish from the polluted Elbe estuary. The use of the heteroxenous/monoxenous species ratio as a marker was not useful at the locations investigated because of the dominance of heteroxenous species at all habitats. Since EROD activity and macrophage aggregate activity were dependent on sex and maturity of female flounder, only male fish were taken into consideration for the integrated evaluation of data. All biochemical and histochemical tests were able to reflect accurately the site-specific differences, as well as an observed pollution event at the end of 1995 as determined by chemical analyses. The correlation analysis revealed a connection not only between the single parasitological and biochemical parameters but also within these groups. The non-specific immune response and Trichodina infection intensity were correlated with all other parameters, leading to the assumption that these may serve as links between the lowest and the highest levels of biological organisation. The simultaneous use of metabolic and parasitological results facilitated the interpretation of the observed variations of the data and the distinction between natural variation and pollution-mediated effects. Received: 1 March 1999 / Received in revised form: 25 May 1999 / Accepted: 7 June 1999     

QTL analysis of tolerance to a German strain of BYDV-PAV in barley (Hordeum vulgare L.)

One hundred and forty six barley doubled-haploid lines (DH lines) were tested for variation in grain yield, yield components, plant height, and heading date after artificial infection with a German isolate of barley yellow dwarf virus (BYDV-PAV-Braunschweig). Of these 146 lines 76 were derived from the cross of the barley yellow dwarf virus (BYDV) tolerant cultivar ’Post’ to cv ’Vixen’ (Ryd2) and 70 from the cross of Post to cv ’Nixe’. Phenotypic measurements were gathered on both non-infected plants and plants artificially inoculated with BYDV-PAV by viruliferous aphids in pot and field experiments for three years at two locations. For all traits a continuous variation was observed suggesting a quantitative mode of inheritance for tolerance against BYDV-PAV. Using skeleton maps constructed using SSRs, AFLPs and RAPDs, two QTLs for relative grain yield per plant after BYDV infection, explaining about 47% of the phenotypic variance, were identified in Post × Vixen at the telomeric region of chromosome 2HL and at a region containing the Ryd2 gene on chromosome 3HL. In Post × Nixe, a QTL was found in exactly the same chromosome 2HL marker interval. In this cross, additional QTL were mapped on chromosomes 7H and 4H and together these explained about 40% of the phenotypic variance. QTL for effects of BYDV infection on yield components, plant height, and heading date generally mapped to the same marker intervals, or in the vicinity of the QTL for relative grain yield, on chromosomes 2HL and 3HL, suggesting that these regions are of special importance for tolerance to the Braunschweig isolate of BYDV-PAV. Possible applications of marker-assisted selection for BYDV tolerance based on these results are discussed. Received: 1 December 2000 / Accepted: 9 March 2001     

The effects of kinship and density on growth and metamorphosis of the bronze frog (Rana temporalis) tadpoles

  Five clutches of Rana temporalis eggs collected along a stream were reared separately until formation of Gosner stage 25 tadpoles. The effect of kinship and density was then studied by rearing ten siblings in 1 (sib 1) or 5 (sib 5) l water, and mixed rearing of ten tadpoles (2 from each of the 5 clutches) in 1 (mix 1) or 5 (mix 5) l water; each group replicated five times. In all the groups tadpoles showed a sigmoid growth curve. Both kinship and density interacted to affect mean proportions of individuals reaching metamorphic climax (MC), mean body mass, and size frequency at MC (day 79). The proportion of tadpoles reaching MC was highest in sib 5 (82%) followed by mix 5 (77%), sib 1 (73%), and mix 1 (64%) groups. Crowding plus mixing significantly lowered the median developmental stage mean body mass and broadened the spectrum of developmental stages or size classes at MC. The size difference of individuals at MC was inversely related to density of rearing. Frequency of different developmental stages was comparable in sib 1, sib 5, and mix 5 groups. Interestingly, small individuals were significantly greater in number in the mix 1 group compared to sib 1. The above findings suggest that genotypic heterogeneity in conjunction with crowding retards growth rate of tadpoles in comparison with those reared in a genetically homogenous (sib) environment. Further, they suggest that the adverse effect of mixed rearing is context dependent. Received: 12 April 1999 / Received in revised form: 16 June 1999 / Accepted: 18 June 1999     

Plasma androgens and aggressiveness in captive winter white-throated sparrows (Zonotrichia albicollis)

Captive white-throated sparrows (Zonotrichia albicollis), held under a natural photoperiod, were pair tested to evaluate dominance and aggressiveness in relation to plasma androgen levels and plumage morphology. Androgen levels were determined in November, 1 week before behavioral testing began; in January, at the conclusion of behavioral testing; and again in March. November plasma androgen levels of captive birds did not differ from those of a group of feral birds and were greater than androgen levels in January and March. Androgen levels of male and female white-striped and tan-striped birds did not differ for any collection date. November plasma A levels were significantly positively correlated with the display of aggressive behaviors by test birds and were inversely correlated, though not significantly, with hierarchy position. Plasma androgen levels determined in January and March were not related to aggressiveness or dominance. These data suggest that circulating androgens and morphological characters may act synergistically, at the onset of flock formation, to influence avian winter dominance hierarchies.     

An integrative approach to sex change: social, behavioural and neurochemical changes in Lythrypnus dalli (Pisces)

This study examined three aspects of protogynous sex change in Lythrypnus dalli (Gobiidae): (1) social influences on the rate of sex change, (2) the sequence of behavioural changes, and (3) neuroendocrine changes. Social groups consisted of either four females, or four females with a male who was subsequently removed. Sex change occurred most rapidly in male- removed groups when the sex changer was larger than other females. Sex changers in female only groups and sex changers not larger than other females in male-removed groups changed sex at similar rates. These differences may be explained by two factors that affect dominance: prior knowledge of the social group and greater size. Sex changers were dominant to other females prior to male removal, and larger sex changers increased displacement rates three-fold immediately after male removal. Sex changers in the other groups did not show this increase in displacements. This early establishment of dominance accounts for the overall difference in the rate of sex change. Prior to spawning, however, all sex changers increased displacements and performed male-typical displays. Arginine vasotocin-immunoreactive forebrain cells of sex changers were similar in size to field-collected males, and larger than field-collected females. Previously nesting males also changed sex in male-only groups, but at slow rates. These data are combined with those of existing studies to generate an integrative model of sex change in this goby. Received: 17 March 1999 / Received in revised form: 15 May 1999 / Accepted: 28 May 1999     

Androgen receptor immunoreactivity in the male and female Syrian hamster brain

To investigate potential mechanisms for sex differences in the physiologic response to androgens, the present study compared the hormonal regulation of intracellular androgen receptor partitioning and the distribution of androgen receptor immunoreactivity in select brain regions from male and female hamsters. Androgen receptors were visualized on coronal brain sections. Two weeks after castration, androgen receptor immunoreactivity filled the neuronal nuclei and cytoplasm in males and females. In gonad‐intact males and females, androgen receptor immunoreactivity was limited to the cell nucleus. Whereas exogenous dihydrotestosterone prevented cytoplasmic immunoreactivity, estrogen at physiologic levels did not. These results suggest that nuclear androgen receptor immunoreactivity in gonad‐intact females is maintained by endogenous androgens, and that androgens have the potential to influence neuronal activity in either sex. However, sex differences in the number and staining intensity of androgen‐responsive neurons were apparent in select brain regions. In the ventral premammillary nucleus, ventromedial nucleus of the hypothalamus, and medial amygdaloid nucleus, androgen receptor staining was similar in gonadectomized males and females. In the lateral septum, posteromedial bed nucleus of the stria terminalis (BNSTpm), and medial preoptic nucleus, the number of androgen receptor–immunoreactive neurons was significantly lower in females (p < .05). Moreover, the integrated optical density/cell in BNSTpm was significantly less in females (1.28 ± 0.3 units) than in males (2.21 ± 0.2 units; p < .05). These sex differences in the number and staining intensity of androgen‐responsive neurons may contribute to sex differences in the behavioral and neuroendocrine responses to androgens. © 1999 John Wiley & Sons, Inc. J Neurobiol 39: 359–370, 1999     

Molecular cloning of ABNORMAL FLORAL ORGANS: a gene required for flower development in Arabidopsis

 We report the cloning and characterization of the gene ABNORMAL FLORAL ORGANS (AFO), which is required for normal flower development in Arabidopsis. afo mutant flowers show defects in all four floral whorls. The number of organs in each whorl varies. Most flowers consist of reduced numbers of petals and stamens, even though supernumerary sepals and carpels may be observed. Abnormal organ structure is evident from an early stage. Mosaic first whorl organs are common, with some sepals taking on petaloid or staminoid characteristics. Stamens are often deformed, having thin filaments and reduced anthers, yet occasionally producing viable pollen. Partial fertility is indicated by some seed setting. The afo-1 mutation is caused by insertion of a gene trap Ds transposable element. The AFO gene was cloned and is predicted to encode a novel protein of 229 amino acids. The expression of AFO mRNA by northern blot analysis in combination with mutant phenotype suggests that the AFO gene product plays an important role in Arabidopsis flower development. We also report that antherless, a previously described male-sterile mutation, is allelic to afo-1. Received: 3 September 1998 / Revision accepted: 15 December 1998     

Factors affecting vigilance in springbok: importance of vegetative cover, location in herd, and herd size

Vigilance in vertebrates is often inversely related to group size. We present evidence that distance to bushes and location within the herd are also critical factors in vigilance in springbok (Antidorcas marsupialis) in Etosha National Park, Namibia, where they are the preferred prey of cheetahs (Acinonyx jubatus). Most springbok feed in heterospecific herds, both by grazing on grass and browsing on bushes. We studied 1245 animals; variations in vigilance (time alert) were explained by location within the herd, distance to bushes and roads, number of springbok in each herd, and gender and age. Vigilance time decreased with increasing herd size, with increasing distance to bushes and roads, and with density. Springbok on the edge of herds devoted significantly more time to vigilance than did those in other locations, and vigilance in edge animals decreased with group size. Adults were more vigilant than young, and males were more vigilant than females. Position within the herd, and distance from bushes, were the most important variables influencing vigilance. Location in the herd and gender/age affected both browsing and grazing springbok, although other factors accounted for the differences in vigilance between browsing and grazing springbok: 1) group size was not significant for browsers, but it was for grazers, and 2) distances to bushes and road were not significant for browsers, but they were for grazers. These data relate to the risk from predators and the benefits from other group members. Springbok in bushes cannot see all members of the herd, cannot derive early warning from many group members, and are more at risk from predators because the latter can hide in the bushes. Received: 17 May 1999 / Received in revised form: 30 August 1999 / Accepted: 29 November 1999     

A combined stereo-photogrammetry and underwater-video system to study group composition of dolphins

One reason for the paucity of knowledge of dolphin social structure is the difficulty of measuring individual dolphins. In Hector’s dolphins, Cephalorhynchus hectori, total body length is a function of age, and sex can be determined by individual colouration pattern. We developed a novel system combining stereo-photogrammetry and underwater-video to record dolphin group composition. The system consists of two downward-looking single-lens-reflex (SLR) cameras and a Hi8 video camera in an underwater housing mounted on a small boat. Bow-riding Hector’s dolphins were photographed and video-taped at close range in coastal waters around the South Island of New Zealand. Three-dimensional, stereoscopic measurements of the distance between the blowhole and the anterior margin of the dorsal fin (BH-DF) were calibrated by a suspended frame with reference points. Growth functions derived from measurements of 53 dead Hector’s dolphins (29 female : 24 male) provided the necessary reference data. For the analysis, the measurements were synchronised with corresponding underwater-video of the genital area. A total of 27 successful measurements (8 with corresponding sex) were obtained, showing how this new system promises to be potentially useful for cetacean studies. Received: 5 January 1999 / Accepted: 29 June 1999     

Distribution patterns of bacterioplankton and chlorophyll-a in the German Wadden Sea

 In a first synoptic evaluation, the temporal and spatial distribution of bacterioplankton and chlorophyll-a were determined in the German Wadden Sea. Three surveys were undertaken in winter, spring, and summer of 1994 using up to eight ships simultaneously between the river Ems and Sylt island. Despite intensive hydrodynamic mixing of the Wadden Sea water, spatial gradients were obvious. The abundance of bacterioplankton ranged from 0.4 to 26×10⁵ ml^–1 and chlorophyll-a varied between <0.1 and 79 μg l^–1. In winter, relatively homogeneous distribution patterns of both parameters with small gradients were found. Highest chlorophyll-a values connected with a highly patchy structure were observed in spring, while in summer both total chlorophyll-a values and the complexity of the distribution pattern had decreased. In contrast, bacterial numbers increased steadily from January to July with the highest bacterial densities and greatest patchiness observed in summer. Moreover, in some regions of the Wadden Sea, a trophic succession of algae as carbon producers and bacteria as consumers was evident. Correlation analysis verified the relationship between bacteria and chlorophyll a, indicating bottom-up control of bacterial abundance in the northern part of the German Wadden Sea. Since the observed regression slope is remarkably low (0.12–0.46) compared to literature values (0.5–0.8), we suggest that the link between phytoplankton and bacteria found here is a special characteristic of the Wadden Sea as a transition zone between the coastal region and the outer North Sea. Received: 26 May 1998 / Accepted:12 November 1998     

Isolation and characterization of two acyl-CoA-binding proteins from proembryogenic masses of Digitalis lanata Ehrh.

 Two acyl-CoA-binding-protein (ACBP) isoforms were isolated from proembryogenic masses of Digitalis lanata Ehrh. by column chromatography and preparative HPLC. The ACBPs had molecular masses of 9926 and 9997 Da, respectively. Partial sequence data indicated high similarity to each other and to ACBPs of other plant species such as Ricinus communis, Brassica napus and Arabidopsis thaliana. The isolated ACBPs bound palmitoyl-CoA with high affinity as determined by isoelectric-point shift. Received: 29 May 1999 / Accepted: 28 August 1999     

Mounting an immune response correlates with decreased androgen levels in male peafowl, <Emphasis Type="Italic">Pavo cristatus</Emphasis>

Studies testing the “immunocompetence handicap hypothesis” have focussed on the immunosuppressive effects of androgens. Several recent studies have reported that mounting a humoral immune response might also result in a decrease in circulating androgen levels via a “negative feedback” on the hypothalamus–pituitary–gonadal axis (HPG). The aim of this correlative study was to analyse these immunosuppressive and HPG-suppressive interactions in reproductively active males of the peafowl. We collected blood samples of free living birds before and after challenging the immune system with a non-pathogenic antigen (sheep erythrocytes), and analysed immune parameters and plasma levels of the two main androgens in birds, testosterone and dihydrotestosterone. Males displaying larger versions of the main secondary sexual trait, the long and conspicuously ornamented train, tended to have higher androgen levels and significantly lower circulating levels of leukocytes, indicating that exaggerated ornaments might signal properties of the endocrine and immune system. Actual circulating levels of androgens did not correlate with the plasma levels of leukocytes and the antibody response to SRBC. However, changes in plasma levels of both androgens showed negative correlation with both leukocytes (P < 0.1) and SRBC responses (P < 0.05). The data therefore support the prediction that activity of the immune system is HPG-suppressive. Such suppression has been proposed to be especially costly during the reproductive season, during which androgens facilitate the expression of exaggerated traits that play an important role in sexual competition.     

A survey of proteinases active during meiotic development

Microsporogenesis in Lilium longiflorum Thunb. is a naturally synchronous process and affords a system in which to study stage-specific events of meiosis and anther development. Zymogram gel analyses were conducted with extracts from a variety of stages of anther development to identify proteinases which likely play roles in anther metabolism. These experiments revealed that several proteinases are present at different stages of anther development, and class-specific inhibitors were used to classify these enzymes. Proteolytic activities increased as anther development proceeded and these activities were temporally correlated with the apoptotic events which precede dehiscence, as well as with events crucial for the maturation of viable pollen. Received: 12 October 1999 / Accepted: 13 November 1999     

Predicting days of high allergenic risk during Betula pollination using weather types

The aim of this study was to build up a picture of the influence of meteorological conditions on pollen and pollinosis, taking account of weather types, pollen concentrations in the air and pollinosis symptoms, with the aim of preventing allergic responses. The study took place in Burgundy from 1996 to 1998, during the pollination of the birch (Betula), which is the most important arborean allergen in this region. We used daily pollen data from four Hirst volumetric traps, identified weather types by Bénichou’s classification, and obtained data on the occurrence of rhinitis, conjunctivitis, asthma and coughing from a sample of 100 patients. These data were analysed by multiple-component analysis. The results show that pollen dispersal is favoured by windy conditions, low relative humidity, precipitation below 2 mm and temperatures above 6°C. Such weather also favours pollinosis, but other particular meteorological situations, even if they do not assist pollen dispersal, can act directly on the development of symptoms: a decrease of temperature (3°C) led to the development of rhinitis and conjunctivitis, while strong winds were associated with many cases of conjunctivitis and asthma, owing to the irritant effect of cold or wind; asthma was favoured by temperature inversions with fog, probably because such weather corresponds to high levels of pollution, which act on bronchial hyperreactivity. Because the weather types favouring pollination and pollinosis are predicted by the meteorological office, this can constitute a tool for reducing the effect of high-risk allergenic days. Received: 12 December 2000 / Revised: 24 April 2001 / Accepted: 9 May 2001     

Production of triploid plants of mulberry (Morus alba L) by endosperm culture

 A continuously growing callus was obtained from immature endosperm of Morus alba L Cv S-36 cultured on Murashige and Skoog medium containing 5 μm 2,4-dichlorophenoxyacetic acid. Shoot buds were produced when the callus was subcultured on a medium containing a cytokinin or a cytokinin and 1-naphthaleneacetic acid (NAA). The maximum number of shoots was formed on the medium containing thidiazuron (1 μM), or benzylaminopurine (5 μM) and NAA (1 μM). Shoots were multiplied by forced axillary branching and rooted in vitro. Endosperm-derived plants were established in soil. Each of the ten plants examined cytologically was triploid (3 n=42). Received:17 February 1999 / Revision received: 4 May 1999 / Accepted: 19 May 1999     

Cryopreservation and plant regeneration from somatic embryos of Pinus patula

 Embryogenic tissue of Pinus patula Scheide et Deppe was cryopreserved for 8 weeks using sorbitol and dimethylsulfoxide (DMSO) as cryoprotectants. Results indicate that 0.3 M sorbitol and 5% DMSO had the best cryoprotecting effect. The recovered tissue initially underwent a lag phase but then continued to proliferate normally on MSG3 maintenance medium. Recovered tissue was placed onto MSG5 maturation medium, and embryos were isolated and germinated. Plantlet regeneration from the recovered tissue was achieved. Received: 16 April 1999 / Revision received: 26 July 1999 / Accepted: 17 August 1999