Changes in life‐history traits in an expanding moss species: phenotypic plasticity or genetic differentiation? A reciprocal transplantation experiment with Pogonatum dentatum

A transplant experiment was performed to investigate whether differences in life-history traits of the bryophyte Pogonatum dentatum that recently expanded its distribution range, were genetically or environmentally determined, or a combination of both. Plants were transplanted reciprocally between the original mountain area and a recently colonised lowland area. Vegetative biomass of annual segments and branches tended to be higher in the mountain area than in the lowland area. Reproductive investment was higher for plants transplanted to the lowland area, and lowland shoots tended to produce larger sporophytes than mountain shoots when placed in the same environment. Age of reproducing shoots showed no consistent pattern.
Native shoots were often outperformed by non-native shoots transplanted into the same site. Non-native shoots grew larger and produced larger sporophytes than native shoots. Much of the observed variation was at the site level instead of between mountain and lowland areas, with both genetic origin and environmental effects contributing together. Range expansion of P. dentatum may have taken place by dispersal from populations with shoots whose growth is plastic. Such shoots grow larger and potentially produce more spores for dispersal.     

Genetic variation and structure in the expanding moss Pogonatum dentatum (Polytrichaceae) in its area of origin and in a recently colonized area

Genetic variation in the expanding moss species Pogonatum dentatum was studied using intersimple sequence repeat (ISSR) markers. The genetic consequences of range expansion were studied by comparing source populations in a mountain area with populations from a recently colonized lowland area in Sweden. Indices of genetic variation show slightly lower number of alleles per locus in the lowlands and a similar gene diversity in both areas. Three of four lowland populations had evidence of a recently passed bottleneck. Considerably higher haplotype diversity was found in the recently colonized lowlands compared to source populations in the mountains. Patterns of allelic diversity suggest that P. dentatum experiences loss of genetic variation through founder effects and genetic drift when expanding its distribution range. Higher haplotypic diversity, less linkage disequilibrium, and fewer compatible loci indicate that sexual recombination is relatively more important in the lowlands compared to the mountains. A likely explanation is higher success of establishment from spores in the lowlands, while clonal propagation predominates in the mountains. Less genetic differentiation among lowland populations indicates more gene flow in the lowland area, involving more spores and/or fragments moving among populations.     

Leaf‐litter colonisation and breakdown in relation to stream typology: insights from Mediterranean low‐order streams

1. Scant information is available on leaf breakdown in streams of arid and semiarid regions, including the Mediterranean, where environmental heterogeneity can be high and the relationship between stream characteristics and leaf breakdown is poorly known. We tested the hypotheses that differences in leaf breakdown metrics would be substantially higher between mountain and lowland Mediterranean streams than among streams within each subregion and that variability among streams would be substantially higher in the lowlands, because permanent reaches in the semiarid lowland streams are rare and isolated. 2. We compared leaf breakdown and associated dynamics of nutrients, fungi and invertebrates in low‐order Mediterranean streams draining sub‐humid forests in the Sierra Nevada Mountains and nearby semiarid lowlands of south‐eastern Spain. Streams differed between the two subregions mainly in water ion content, temperature and riparian tree cover. We detected higher environmental heterogeneity among streams within the lowlands compared to the Sierra Nevada mountain range. In the lowlands, breakdown coefficients (k) of alder leaves spanned almost the entire range reported for this species from temperate streams, overlapping with less variable breakdown coefficients in the Sierra Nevada. 3. The high variability of k values among the lowland sites appeared to be caused primarily by variability in the composition and abundance of a few leaf‐consuming invertebrate taxa, particularly the snail Melanopsis praemorsa. Fungal and nutrient dynamics were less variable among sites within each subregion. 4. These results indicate that the critical condition for stream functional assessment of well‐constrained breakdown rates, or related metrics, could be met at reference sites within homogenous bio‐geo‐climatic regions such as the Sierra Nevada. By contrast, in heterogeneous areas such as the semiarid lowland streams, natural variability of breakdown rates can greatly exceed the magnitude of effects expected in response to anthropogenic disturbances.     

Why on the snow? Winter emergence strategies of snow‐active Chironomidae (Diptera) in Poland

A long‐term study of adult non‐biting midges (Chironomidae) active in winter on the snow in mountain areas and lowlands in Poland yielded 35 species. The lowland and mountain communities differed significantly in their specific composition. The mountain assemblage was found to be more diverse and abundant, with a substantial contribution from the subfamily Diamesinae, whereas Orthocladiinae predominated in the lowlands. Orthocladius wetterensis Brundin was the most characteristic and superdominant species in the winter‐active chironomid communities in both areas. Only a few specimens and species of snow‐active chironomids were recorded in late autumn and early winter. The abundance of chironomids peaked in late February in the mountain and lowland areas with an additional peak in the mountain areas in early April. However, this second peak of activity consisted mainly of Orthocladiinae, as Diamesinae emerged earliest in the season. Most snow‐active species emerged in mid‐ and late winter, but their seasonal patterns differed between the 2 regions as a result of the different species composition and the duration of snow cover in these regions. Spearman's rank correlation coefficient tests yielded positive results between each season and the number of chironomid individuals recorded in the mountain area. A positive correlation between air temperature, rising to +3.5 °C, and the number of specimens recorded on the snow in the mountain community was statistically significant. The winter emergence and mate‐searching strategies of chironomids are discussed in the light of global warming, and a brief compilation of most important published data on the phenomena studied is provided.     

Reproductive modes in the epixylic hepatic Anastrophyllum hellerianum

The frequency and dynamics of sexual and asexual reproduction were investigated in a dioecious epixylic hepatic, Anastrophyllum hellerianum, which has declined in recent decades in Finland as a consequence of forestry practices. In our investigation asexual reproduction by gemmae was the dominant mode of reproduction and specialised gemmiparous shoots were present in all colonies studied. The proportions of dead shoots were considerably higher among sex-expressing than among non-sexexpressing shoots. Our results suggest that lower reproductive investment is required for asexual than for sexual reproduction. For instance, no trade-off is detected between asexual reproduction and survival of the gemmiparous shoots in A. hellerianum. Sexual reproduction occurred only in 12% of the colonies and it was promoted by the following factors: medium shoot density, high proportion of sex-expressing shoots, an even sex ratio and very short distances between individuals representing opposite sexes. The ratio of dead males to dead females was significantly female-biased, which suggests higher mortality among female shoots. At the level of individual shoots, more spores than gemmae were produced. However, as a consequence of the low frequency of sporophyte-bearing shoots, gemma production highly exceeded spore production at the colony level. Furthermore, cultivation tests of the propagules showed that gemmae germinate faster than spores.     

Development and characterization of microsatellite loci in <Emphasis Type="Italic">Ixeridium dentatum</Emphasis> (Asteraceae,Lactuceae)

We isolated and characterized seven microsatellite loci for the perennial herb Ixeridium dentatum ssp. dentatum, an apomictic triploid distributed throughout the lowland areas of East Asia. The number of alleles ranged from two to seven in 32 screened individuals of I. dentatum ssp. dentatum from Japan. The observed and expected heterozygosities were 0.000–0.950 and 0.000–0.891, respectively, calculated using genotypes of 20 individuals of I. dentatum ssp. nipponicum. One locus (msid4) deviated significantly from Hardy–Weinberg equilibrium (P = 0.0001). These microsatellites were also tested for cross-amplification in 11 other taxa of Lactuceae, including five endangered taxa. These primers should be useful genetic tools not only for Ixeridium but also for other Lactuceae taxa.     

Can it be all more simple? Manufacturing aflatoxin biocontrol products using dry spores of atoxigenic isolates of Aspergillus flavus as active ingredients

Aflatoxin contamination of staple crops, commonly occurring in warm areas, negatively impacts human and animal health, and hampers trade and economic development. The fungus Aspergillus flavus is the major aflatoxin producer. However, not all A. flavus genotypes produce aflatoxins. Effective aflatoxin control is achieved using biocontrol products containing spores of atoxigenic A. flavus. In Africa, various biocontrol products under the tradename Aflasafe are available. Private and public sector licensees manufacture Aflasafe using spores freshly produced in laboratories adjacent to their factories. BAMTAARE, the licensee in Senegal, had difficulties to obtain laboratory equipment during its first year of production. To overcome this, a process was developed in Ibadan, Nigeria, for producing high-quality dry spores. Viability and stability of the dry spores were tested and conformed to set standards. In 2019, BAMTAARE manufactured Aflasafe SN01 using dry spores produced in Ibadan and sent via courier and 19 000 ha of groundnut and maize in Senegal and The Gambia were treated. Biocontrol manufactured with dry spores was as effective as biocontrol manufactured with freshly produced spores. Treated crops contained safe and significantly (P < 0.05) less aflatoxin than untreated crops. The dry spore innovation will make biocontrol manufacturing cost-efficient in several African countries.     

In vitro formation of resting spores by the insect pathogenic fungus Entomophaga maimaiga

Field-collected resting spores (azygospores) of the fungal pathogen of Lymantria dispar (gypsy moth), Entomophaga maimaiga, have been used to release this biological control agent in areas where this pathogen is not established. We have found that E. maimaiga can produce resting spores in vitro using Grace's insect tissue culture medium (95%) plus fetal bovine serum (5%). The majority of spores become mature between 7 and 21 days after cultures are initiated. Spore production varies by fungal isolate; of 38 isolates tested, 10 produced no resting spores while 7 produced >1000 resting spores/ml. Resting spore production was not affected when isolates were mixed. Glycerol (used for fungal storage), trehalose, and selected amino acids each inhibited resting spore formation. Fetal bovine serum was required for spore production but the presence of >5% yielded lower resting spore densities. A large surface area:volume ratio (12.5 cm(2):ml versus 4.2 cm(2):ml) was required for abundant formation of resting spores. At present, resting spores have only been produced in small volumes with a maximum of 3 x 10(4) resting spores/ml.     

SPORE ULTRASTRUCTURE OF THE MYXOMYCETE PHYSARUM POLYCEPHALUM

Spores of the true slime mold Physarum polycephalum were examined at several stages of their development by means of scanning and transmission electron microscopy. The spores were globose, spine-covered structures produced within a sporangium enclosed in a tough, noncellular peridium. Cytologically, the spore represented a typical eukaryotic cell, having discrete organelles similar to spores of other myxomycetes. The presence of dictyosomes, helical filaments, and microbodies in these cells, as well as the further elucidation of the cell wall and the “polysaccharide-containing” areas, represent new contributions to the ultrastructure of the myxomycete spore. Of special interest were observations of metaphase nuclei just prior to spore cleavage, interphase nuclei in young spores, and nuclei in mature spores containing synaptonemal complexes. These observations indicate that in Physarum polycephalum mitosis occurs just prior to spore cleavage, and meiosis takes place after spore cleavage.     

Spore production and mycorrhizal development in various tropical crop hosts infected with Glomus clarum

Plant growth, mycorrhizal development and vesicular arbuscular spore production were examined in five tropical crop host species inoculated with Glomus clarum and grown in a glasshouse. In one of the two experiments, sequential harvests of maize, sorghum and chickpea were made in order to study spore production in relation to plant growth and mycorrhizal development. Spore numbers in each of these hosts increased at a fairly constant rate until maximum plant dry weight, when spore production ceased. Sorghum and maize produced considerably more spores than chickpea, with spore numbers being closely correlated with mycorrhizal root length. In the second experiment, Glomus clarum was cultured on each of maize, millet, sorghum, groundnut and chickpea for three consecutive generations before cross-inoculation of the spores from each host onto all five hosts. Sporulation with respect to host size was generally greatest when the inoculum used to infect a host had been produced on that host. The growth-promoting effects of the fungus were not influenced by the source of the inoculum. More spores were produced on the cereals than the legumes. Differences in spore numbers amongst hosts and plant generations were apparently influenced mainly by infected root length and by the growth period.     

The impact of nutrition on spore yields for various fungal entomopathogens in liquid culture

Spore yields were measured for various fungal entomopathogens grown in six nutritionally different liquid media with low and high carbon concentrations (8 and 36 g l^–1, respectively) at carbon-to-nitrogen (C:N) ratios of 10:1, 30:1 and 50:1. Six fungi were tested: two Beauveria bassiana strains, three Paecilomyces fumosoroseus strains and one Metarhizium anisopliae strain. Spore yields were examined after 2, 4 or 7 days growth. In general, highest spore yields were obtained in media containing 36 g/l and a C:N ratio of 10:1. After 4 days growth, highest spore yields were measured in the three Paecilomyces isolates (6.9–9.7 × 10⁸ spores ml^–1). Spore production by the B. bassiana isolates was variable with one isolate producing high spore yields (12.2 × 10⁸ spores ml^–1) after 7 days growth. The M. anisopliae isolate produced low spore concentrations under all conditions tested. Using a commercial production protocol, a comparison of spore yields for the coffee berry borer P. fumosoroseus and a commercial B. bassiana isolate showed that highest spore concentrations (7.2 × 10⁸ spores ml^–1) were obtained with the P. fumosoroseus isolate 2-days post-inoculation. The ability of the P. fumosoroseus strain isolated from the coffee berry borer to rapidly produce high concentrations of spores prompted further testing to determine the desiccation tolerance of these spores. Desiccation studies showed that ca. 80% of the liquid culture produced P. fumosoroseus spores survived the air-drying process. The virulence of freshly produced, air-dried and freeze-dried coffee berry borer P. fumosoroseus blastospores preparations were tested against silverleaf whiteflies (Bemisia argentifolii). While all preparations infected and killed B. argentifolii, fresh and air-dried preparations were significantly more effective. These results suggest that screening potential fungal biopesticides for amenability to liquid culture spore production can aid in the identification of commercially viable isolates. In this study, P. fumosoroseus was shown to possess the production and stabilization attributes required for commercial development.     

Application of response surface methodology in medium optimization for spore production of <Emphasis Type="Italic">Coniothyrium minitans</Emphasis> in solid-state fermentation

Summary Spore production of Coniothyrium minitans was optimized by using response surface methodology (RSM), which is a powerful mathematical approach widely applied in the optimization of fermentation process. In the first step of optimization, with Plackett–Burman design, soluble starch, urea and KH²PO⁴ were found to be the important factors affecting C. minitans spore production significantly. In the second step, a 2³ full factorial central composite design and RSM were applied to determine the optimal concentration of each significant variable. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the critical components for the maximum were obtained as follows: soluble starch 0.643 (36.43 g. l⁻¹), urea −0.544 (3.91 g l⁻¹) and KH²PO⁴ 0.049 (1.02 g l⁻¹) with a predicted value of maximum spore production of 9.94 × 10⁹ spores/g IDM. Under the optimal conditions, the practical spore production was 1.04 × 10¹⁰ spores/g IDM. The determination coefficient (R²) was 0.923, which ensure an adequate credibility of the model.     

Structural and developmental studies of Chlamydomyzium oviparasiticum from Rhabditis nematodes and in culture

The oomycete (Peronosporomycete) Chlamydomyzium oviparasiticum, previously recorded as a parasite of rotifer eggs, was found infecting Rhabditis nematodes in a sample of rotting garden compost. For the first time C. oviparasiticum was cultured in liquid media, which enabled more detailed studies of zoospore behaviour and facilitated the use of confocal microscopy. Rhabditis nematodes were successfully re-infected from liquid-cultured inoculum. Light (including video) microscopy and transmission electron microscopy were used to document details of thallus development, zoospore release and resting spore morphology to enable comparison with other oomycete species. This species showed several significant saprolegnialian characters such as the ‘achlyoid’ pattern of spore formation, centrifugal cleavage and structured encystment vesicles. In contrast, spore release into a transient vesicle was a peronosporalean characteristic. The thick-walled resting spores showed relatively poor cytoplasmic preservation and had a thick multi-layered wall. It was still not possible to unequivocably decide whether these were chlamydospores or parthenogenically formed oospores. The phylogenetic significance of these observations is discussed.     

EXCHANGES OF INFORMATION,ENERGY & MATERIALS IN SYMBIOSES

We investigated patterns of spore dispersal in the giant kelp Macrocystis pyrifera by collecting 80 independent measurements of spore dispersal from isolated individuals and isolated groups of individuals over a two‐year period. Our results indicate that giant kelp spores routinely disperse both short (i.e. a couple meters) and long (i.e. hundreds to thousands of meters) distances depending on the oceanographic conditions. One consequence of spore dispersal over short distances is self‐fertilization (i.e., fertilization between male and female gametophytes derived from the same sporophyte). Field experiments designed to test the effects of self‐fertilization on lifetime fitness in Macrocystis revealed significant inbreeding depression. Birth rates in self‐fertilized populations were ca. 50% of those produced from outcrossing, which lead to significant differences in cohort size that persisted up through the adult stage. In contrast to outcrossed populations, very few individuals produced from selfing became reproductively mature, and those that did were significantly less fecund than outcrossed individuals. By contrast, long‐range dispersal of spores leads to increased rates of outcrossing. However, long‐range dispersal is typically accompanied by massive dilution of spores, leading to low densities of spore settlement. Sparse spore settlement decreases the overall chance of fertilization in the microscopic gametophyte generation thereby reducing the potential for colonization of the macroscopic sporophyte stage. Large population size of adult sporophytes coupled with the synchronous release of spores in response to environmental cues can help offset the effects of spore dilution and extend the distances over which giant kelp is able to colonize.     

Reproductive attributes of <Emphasis Type="Italic">Campanula</Emphasis> populations from Mt Olympos,Greece

We studied five reproductive attributes (fruit production and volume, seed content and weight, and seed germination) of different populations of Campanula species occurring along the elevation gradient of Mt Olympos, the highest mountain of Greece (2917 m), in order to examine whether and how they change with elevation, and if there are trade-offs among them. We also studied flower and fruit (reproductive) losses and population features in order to estimate whether there might be a threat to the survival of Campanula species on Mt Olympos. The genus is represented by nine species, ranging from cosmopolitan to steno-endemic. All species were found non-autogamous, and hence they depend on pollinators for their sexual reproduction. At the genus level, there is a trade-off between seed content per fruit and seed weight. At the species level, this is manifested only for C. glomerata, which produces the heaviest seeds, far exceeding all other species, and for C. spatulata, represented on Mt Olympos by two elevation distinct subspecies. Changes of the reproductive attributes are not continuous all along the elevation gradient at the genus level nor are they monotonic at the species level. Nevertheless, upland Campanula populations (≥1500 m) produce fewer fruits per individual, containing fewer but heavier seeds than lowland populations. This indicates a transition from ‘r’ to ‘K’ strategies, from low to high elevations, respectively. The patterns of change of reproductive attributes indicate prevalence of phylogenetic over environmental constraints. Laboratory tests showed that for many populations germinability is high at 15 °C, but there are both inter- and intraspecific differences regarding temperature optima. Ecotypic variation among populations of the same species at different elevations was evident; populations from higher elevations had lower optima and vice versa. Reproductive losses, ranging from zero to over 90%, occurred mainly at the stage of fruit maturation; grazing by sheep and goats was the usual cause. Though such losses might affect the short-lived species (the annual C. sparsa and the biennial C. lingulata), which depend entirely on fruit maturation and seed dispersal for their reproduction, there does not seem to be a serious threat to the survival of Campanula species on the mountain.     

Long term trends in outdoor <Emphasis Type="Italic">Aspergillus/Penicillium</Emphasis> spore concentrations in Derby,UK from 1970 to 2003 and a comparative study in 1994 and 1996 with the indoor air of two local houses

Aspergillus/Penicillium spore concentrations have been monitored in Derby since 1970 using a volumetric spore trap, with full year data from 1991. In addition a short comparative study with the indoor air was undertaken at two local houses in 1994 and 1996. Aspergillus/Penicillium spores were present in the Derby air throughout the year and often reached maximum monthly cumulative concentrations in the autumn, although they were occasionally the dominant spores in the winter when total spore concentrations were low. Very high daily concentrations could occur at any time of year with a count of over 5000 recorded. Peak days in the autumn and winter of 2002–2003 were examined on a two hourly basis showing higher concentrations in the middle of the day. There was a positive correlation of cumulative monthly Aspergillus/Penicillium totals with maximum temperature. Indoor data from the two houses was examined on a daily basis and compared with simultaneously sampled outdoor daily spore concentrations. The elevated Aspergillus/Penicillium spore levels found in the older of the two houses occurred on all of the days sampled. Compared to the modern house, the Aspergillus/Penicillium spore concentrations in the old house represented a much higher percentage of the total spore count than in the modern one. The correlation between outdoor Aspergillus/Penicilliumspore concentrations and the indoor air of the old house was 0.62, whereas in the modern house it was 0.31. Peak hourly samples of Aspergillus/Penicillium spore counts occurred at times of greatest activity.     

Development of <Emphasis Type="Italic">in vitro</Emphasis> methods for <Emphasis Type="Italic">ex situ</Emphasis> conservation of <Emphasis Type="Italic">Eucalyptus impensa</Emphasis>, an endangered mallee from southwest Western Australia

A method is described for in vitro propagation of the critically endangered ‘Eneabba mallee’ (Eucalyptus impensa) from southwest Western Australia. Half-strength MS medium supplemented with 0.25 μM 6-benzylaminopurine and 2.5 μM kinetin resulted in the best combination of shoot multiplication and shoot quality compared to other treatments. Shoots of this species tended to be very compact under in vitro conditions. Shoot length was significantly enhanced with the addition of 0.5 or 1.0 μM gibberellic acid (A₄ isomer) when compared to basal medium (no hormone supplements) or basal medium containing only cytokinin (0.5 μM zeatin). Up to 97.0 ± 3.0% of shoots produced roots on 1/2 MS medium supplemented with a combination of 5 μM indolebutyric acid and 0.5 μM α-naphthaleneacetic acid. Over 70% of shoots transferred to potting mixture remained viable after 3 months. This study has significantly progressed ex situ conservation initiatives for Eucalyptus impensa.     

Southern Scandinavian lowland populations of Rhytidium rugosum (Hedw.) Kindb. (Bryophyta,Rhytidiaceae) differ significantly from those in the mountains

Abstract

Haplotype diversity patterns based on internal transcribed spacer (ITS) and gpd (glyceraldehyde 3-phosphate dehydrogenase) in Scandinavian Rhytidium rugosum were compared between four populations from the mountain range and two in the southern lowlands. The mountain populations differ from the southern lowland ones in haplotype composition and the lowland ones differ from each other, suggesting different histories. A coalescent-based analysis (LAMARC) indicated immigration into the population now found in the mountains from that represented in the lowlands. Because several factors argue against such a dispersal scenario within the relatively short postglacial period, it is proposed that these migration patterns are older than the postglacial period. It is suggested that the mountain range populations immigrated from a source population either in the north-east or within Scandinavia, whereas the lowland populations are likely to represent remains of the glacial population of lowland Europe. The highest haplotype diversity in Scandinavia was found in the Gudbrandsdalen area in southern Norway. This is probably explained by the fact that this is the only region in Europe where R. rugosum regularly reproduces sexually. The lowest diversity was found on the Great Alvar on the island of Öland off the Baltic coast of southern Sweden, and is most likely a result of gradual loss of haplotypes over time.     

Apomixis in the interspecific triploid hybrid fern <Emphasis Type="Italic">Cornopteris christenseniana</Emphasis> (Woodsiaceae)

Cornopteris christenseniana is a "sterile" interspecific triploid hybrid of diploid C. crenulatoserrulata and tetraploid C. decurrenti-alata. Morphological and cytological studies show that, of 41 young plants of Cornopteris that have been propagated naturally in the Fern Garden of the Botanical Gardens, University of Tokyo, 30 plants are the sterile C. christenseniana, 10 are fertile C. decurrenti-alata and 1, fertile C. crenulatoserrulata. This proportion supports the view that the young plants of C. christenseniana are derived from spores of reproductively mature plants of the species cultivated. Cytogenetic observations and culture experiments show that C. christenseniana produces normal spores in various proportions in some sporogenetic pathways that are aberrant from the ordinary process in sexual and apomictic ferns. Under culture conditions, normal spores germinate in rough proportion to the frequency of normal spores, and sporophytes are apogamously produced in rough proportion to the frequency of spore germination. As a whole, the rates of spore germination and apogamous sporophyte development vary according to the specific plant. Taken together, these observations suggest that C. christenseniana is an incipient apomict. Electronic Publication     

Sex ratio and sex expression in an urban population of the silver moss,Bryum argenteum Hedw.

ABSTRACT

Introduction. A well-supported pattern among dioicous bryophytes is male rarity. However, few assessments of bryophyte sex ratios have been made across environmental gradients to assess the role of environment in shaping population sex ratios.

Methods. We systematically surveyed 200 shoots from a 20?m² urban population of Bryum argenteum, and regenerated each shoot apex until sex expression occurred (up to 315 days).

Key results. Female shoots outnumbered male shoots 132 to 68, giving a sex ratio of 1.94♀: 1♂. The female bias was found in two transects in higher light environments but not in the third transect, which had a lower light level and an equal sex ratio. Female shoots took longer than male shoots to reach gametangial induction (122 vs. 60 days) and longer to produce 5 inflorescences (120 vs. 80 days). Male shoots produced an average of 10× the total number of inflorescences compared to female shoots (34 vs. 3.5 inflorescences). Despite producing more inflorescences, male plants also produced more regenerant shoots, thus contradicting the prediction that a higher prefertilisation reproductive effort in males trades off with vegetative proliferation. Female plants harboured significantly more associated microbes than male plants.

Conclusions. Our results support the role of light in influencing sex ratios in this species, suggest that trade-offs between reproduction and vegetative growth may not be strong for males, and indicate a potential role of a sex-specific microbiome in influencing sex ratios.