The genesis of secondary reproductive structures in streptomycete colonies as revealed by confocal microscopy

Streptomycete colonies were examined by confocal microscopy to elucidate the origin of the secondary structures that form the concentric rings of aerial mycelium. These reproductive structures but no the vegetative ones were found to luminesce upon safranine staining. It is shown that the secondary reproductive structures emerge within the substrate mycelium by lateral branching or apical extension of vegetative hyphae, further developing into numerous secondary branches filled with protospores.     

The control of fruiting body formation in the ascomycete Sordaria macrospora Auersw. by regulation of hyphal development

The morphological effects of biotin and L-arginine on fruiting body formation of the ascomycete Sordaria macrospora are investigated by scanning electron and light microscopy. Biotin is recognized as an elongation factor and arginine as a branching factor in vegetative and reproductive hyphae. In the absence of exogenous biotin, development is blocked after the ascogonium-core hypha stage of protoperithecial morphogenesis, whereas linear growth of the myceliar front is maintained. The addition of exogenous arginine to a biotin deficient culture induces the formation of numerous side branches even in the older mycelium. Fruiting body formation, however, remains blocked at the protoperithecial stage as before, because of the inability of the side branches to elongate. When biotin and arginine are administered simultaneously, a most vigorous branching and growth are induced in the older mycelium, accompanied by a rapid and maximal formation of fruiting bodies. The results are summarized in a model of the exogenous control of hyphal morphogenesis. The model is designed to explain the relationship between fruiting and hyphal density as well as the edge effect on fruiting body formation.     

Photoperiodic regulation of receptacle induction in Sargassum horneri (Phaeophyceae) using clonal thalli

We developed a clonal culture of Sargassum horneri to investigate the effect of photoperiod on reproduction in this species. Regenerated vegetative thalli were obtained using lateral branches excised from a thallus grown from a single embryo under short‐day conditions (SD = 10:14 h light : dark cycle). Lateral branches excised from the SD‐regenerated thallus became vegetative thalli that remained in that phase as long as they were cultured under SD. When an excised lateral branch was cultured under long‐day conditions (LD = 14:10 h light : dark cycle), it began to enter the reproductive phase while still less than 50 mm long. Induction of the reproductive phase was accompanied by a distinctive morphological change – suppression of blade formation at the apical region of the branch; elongation of branches without blades was then followed by differentiation of receptacles bearing conceptacles on their surface. Apices of receptacles were able to interconvert between reproductive and vegetative phases, as blades resprouted upon transfer from LD to SD. The critical day length for induction of receptacle formation was between 13 and 14 h; receptacle formation was also induced under SD conditions with night breaks (NBs). These results strongly suggest that reproductive regulation of S. horneri is a photoperiodic long‐day response. NBs with blue and green light were effective for reproductive induction but not with red light. This suggests that blue‐ and/or green‐light photoreceptors are involved in the photoperiodic reproductive response of S. horneri.     

Morphological studies in the tribe Colletieae (Rhamnaceae): analysis of architecture and inflorescences.

A morphological study of vegetative and reproductive structures was carried out on the tribe Colletieae (Rhamnaceae) composed of six genera and 20 species. The plant habit variation, from trees (up to 8 m high) to prostrate shrubs and tangled bushes, suggests structural differences within the group. All the species have an architecture represented by a sequence of monopodial shoots with limited growth, linked sympodially and with acrotonic dichasial ramification. Foliage leaves are decussate and an axillary complex with two serial meristems is associated with each leaf axil. Ancillary shoots (thorn-tipped branches, short shoots, secondary branches and reproductive shoots) are present at the nodes. A single structural plan was observed throughout the tribe. Most of the variability may be ascribed to different strategies for adaptation to environmental conditions, specially xeromorphy. Reproductive structures, in terms of inflorescences, also show conspicuous variation throughout the tribe. Three different models are described; in the first two models individual flowers or 3–7 flowered cymes constitute the paracladia of either deciduous monotelic synflorescences, or of synflorescences proliferating into vegetative long shoots; in the third, uniflorous paracladia arise from perennial short-shoots. Nevertheless, a link within them can be established through steps of truncation, proliferation, reduction and acquisition of rhythmic growth.     

Assay of the inhibitory effect of pentachlorophenol and formaldehyde on mycelial growth and development of reproductive structures inAspergillus niger

The biological activity of the antifungal agents pentachlorophenol (as sodium pentachlorophenolate) and formaldehyde was evaluated by changes in the development of fungal structures following a placement of test strips inoculated withAspergillus niger conidia on a nutrient agar with the toxicant. The method allows a quantitative assessment of biological activity measured as the development of vegetative structures (growth of the mycelium) by metric, and the development of reproductive structures (conidia) ofA. niger by densitometric methods. The use of test strips with dry conidiospores and the evaluation of the results in physical units represent a simple, rapid, exact and inexpensive test of fungitoxic agents.     

Cyclic nucleotides modulate lipoxygenase activity and reproduction in oomycetes

Previous studies in our laboratory demonstrated that epoxy alcohols produced from C₂₀ fatty acids by lipoxygenase activity are associated with maintaining vegetative growth. We have also shown the specific down-regulation of lipoxygenase activity in reproductively competent oomycetes in response to cues which trigger reproduction. Other workers have suggested that cyclic nucleotides may also play a role in the switch between vegetative and reproductive growth of fungi. Reproductive activity was assayed in Achlya americana. Oogonium production was eliminated or reduced in the presence of cAMP, dibutyryl cAMP (0.5 mM), or the phosphodiesterase inhibitors caffeine (0.1, 1 mM) and theophylline (1 mM) and increased by 0.5 mM cGMP. Levels of cAMP were significantly higher in vegetative mycelium and were increased fourfold by exposure to 0.1 mM caffeine. These data suggest that an accumulation of cAMP inhibits reproductive activity while cGMP promotes it. Lipoxygenase activity was determined in the presence of cAMP, cAMP phosphodiesterase inhibitors, and cGMP to determine the interaction between cyclic nucleotides and lipoxygenase activity. Lipoxygenase activity in reproductive mycelium of A. americana or Saprolegnia ferax was reduced compared to vegetative mycelium. Lipoxygenase activity of cultures grown and starved in the presence of cAMP (0.5 mM) or caffeine (0.1 mM) showed significant increases over the comparably treated reproductive controls. Exposure to cGMP had no affect on lipoxygenase activity in A. americana. These data suggest that cAMP may maintain vegetative growth by maintaining relatively high lipoxygenase activity levels while the reproduction promoting effect of cGMP is not via lipoxygenase activity down-regulation. Adenylate cyclase activity was significantly higher in vegetative, compared to reproductive, mycelium of both A. americana and S. ferax. Elevated adenylate cyclase activity in vegetative mycelium supports the hypothesis that cAMP maintains vegetative growth by maintaining high lipoxygenase activity.     

Branch geometry in Cornus kousa (Cornaceae): computer simulations

Computer simulations similar to actual trees were constructed using simple branching rules. Branch orientation with respect to the direction of gravity was a fundamental consideration. In Cornus kousa BUERG. ex HANCE, several types of branches develop from winter buds, varying from orthotropic shoots to plagiotropic ones. Based on actual observations and measurements of branching structures with a wide range of orientations, we made a flexible geometrical model consisting of five forking branches that varied in outgrowth depending on the direction of the shoot with respect to gravity. Repetition of the branching by computer generated a realistic tree pattern, which was close to the shape of a young C. kousa tree. Reproductive shoots seem to be under a branching rule that was a modification of vegetative branching, although the reproductive branch size was considerably smaller than the vegetative one, and reproductive branching was bifurcated instead of five-forked. We conclude that all branchings in orthotropic and plagiotropic shoots in the vegetative phase and shoots in the reproductive phase are formed under the same branching rule, but each has different parameter values.     

Isolation and characterization of differentially expressed genes in the mycelium and fruit body of Tuber borchii

The transition from vegetative mycelium to fruit body in truffles requires differentiation processes which lead to edible fruit bodies (ascomata) consisting of different cell and tissue types. The identification of genes differentially expressed during these developmental processes can contribute greatly to a better understanding of truffle morphogenesis. A cDNA library was constructed from vegetative mycelium RNAs of the white truffle Tuber borchii, and 214 cDNAs were sequenced. Up to 58% of the expressed sequence tags corresponded to known genes. The majority of the identified sequences represented housekeeping proteins, i.e., proteins involved in gene or protein expression, cell wall formation, primary and secondary metabolism, and signaling pathways. We screened 171 arrayed cDNAs by using cDNA probes constructed from mRNAs of vegetative mycelium and ascomata to identify fruit body-regulated genes. Comparisons of signals from vegetative mycelium and fruit bodies bearing 15 or 70% mature spores revealed significant differences in the expression levels for up to 33% of the investigated genes. The expression levels for six highly regulated genes were confirmed by RNA blot analyses. The expression of glutamine synthetase, 5-aminolevulinic acid synthetase, isocitrate lyase, thioredoxin, glucan 1,3-beta-glucosidase, and UDP-glucose:sterol glucosyl transferase was highly up-regulated, suggesting that amino acid biosynthesis, the glyoxylate cycle pathway, and cell wall synthesis are strikingly altered during morphogenesis.     

A GmAOX2b antisense gene compromises vegetative growth and seed production in soybean

The alternative oxidase mediates the cyanide-resistant respiratory pathway in plant mitochondria. In non-thermogenic plants, the role of alternative oxidase in plant growth and development is not well understood. Soybean (Glycine max) lines carrying a GmAOX2b antisense gene had compromised vegetative growth and reproductive performance under typical glasshouse growth conditions. The reduction in vegetative growth was demonstrated by reduction in shoot height, the number of leaves per plant and the green leaf area. Antisense plants also had decreased pod formation and seed to pod ratios, which together led to a reduction in the number and total mass of seed produced. The negative effects of the antisense gene on pod set, seed set, ovule availability and total seed mass were primarily confined to the branches, rather than the main stem. The preferential effect of alternative oxidase suppression in the branches is discussed in relation to the reproductive potential of soybean under stress. Taken together, these results demonstrate that alternative oxidase provides the benefit of sustaining plant vegetative growth and reproductive capacity in soybean.     

Isolation and Characterization of Differentially Expressed Genes in the Mycelium and Fruit Body of Tuber borchii

The transition from vegetative mycelium to fruit body in truffles requires differentiation processes which lead to edible fruit bodies (ascomata) consisting of different cell and tissue types. The identification of genes differentially expressed during these developmental processes can contribute greatly to a better understanding of truffle morphogenesis. A cDNA library was constructed from vegetative mycelium RNAs of the white truffle Tuber borchii, and 214 cDNAs were sequenced. Up to 58% of the expressed sequence tags corresponded to known genes. The majority of the identified sequences represented housekeeping proteins, i.e., proteins involved in gene or protein expression, cell wall formation, primary and secondary metabolism, and signaling pathways. We screened 171 arrayed cDNAs by using cDNA probes constructed from mRNAs of vegetative mycelium and ascomata to identify fruit body-regulated genes. Comparisons of signals from vegetative mycelium and fruit bodies bearing 15 or 70% mature spores revealed significant differences in the expression levels for up to 33% of the investigated genes. The expression levels for six highly regulated genes were confirmed by RNA blot analyses. The expression of glutamine synthetase, 5-aminolevulinic acid synthetase, isocitrate lyase, thioredoxin, glucan 1,3-β-glucosidase, and UDP-glucose:sterol glucosyl transferase was highly up-regulated, suggesting that amino acid biosynthesis, the glyoxylate cycle pathway, and cell wall synthesis are strikingly altered during morphogenesis.     

漆酶同工酶基因在斑玉蕈生长发育过程中的表达分析

为了探明漆酶在斑玉蕈生长发育过程中的功能,对斑玉蕈转录测序预测的13个漆酶基因序列进行分析、鉴定和构建分子系统发育树;检测了不同生长发育时期漆酶的活性和漆酶基因表达水平。研究结果显示：13个基因片段中有10个是漆酶基因。不同的漆酶同工酶之间进化关系存在明显差异,大多数漆酶与木腐菌（金针菇Flammulina filiformis和侧耳属Pleurotus）进化关系较近。对斑玉蕈不同生长发育时期的酶活检测结果显示,从斑玉蕈的菌丝恢复期到钉头期,漆酶活性逐渐升高,而在子实体形成后期酶活逐渐降低。对培养40d、60d和80d的菌丝样品以及不同生长发育时期的样品进RT-qPCR检测,结果显示在菌丝营养生长时期,大多数漆酶基因在第40-60天表达量持续增加1-3倍,而在第60-80天时表达量出现降低的情况。而在生殖生长时期,大多数漆酶基因在转色期或者原基期相对表达量达到最大值,并在子实体期出现降低,这与漆酶活性的检测具有一致性。lcc3、lcc7、lcc8和lcc9在斑玉蕈生殖生长过程中相对表达量出现了10-100倍的上调。这说明从菌丝培养到菌丝扭结形成子实体和子实体发育的过程中,不同的漆酶可能发挥着不同的作用,表达量较高的漆酶基因可能对基质降解和子实体形成起主要作用。     

Origin and ultrastructure of complex septa in Schizophyllum commune development

Summary The mode of origin of cross-walls in living cells of S. commune was studied in basidiospore germinants, vegetative homokaryotic mycelium and the dikaryon of this mushroom. The de novo origin of septa in basidiospore germinants always involved annular ingrowth of cross-walls. The restriction of intercalary growth was observed in the transition from a two-celled hypha to a three-celled unit. Primary branch formation occurred in nonseptate germinants, subterminal cells of the young hypha and in the hyphal apex. Septum formation in purely vegetative homokaryotic mycelium as well as the main cross-wall of the dikaryon was also by annular ingrowth. Ultrastructure studies of the septal pore apparatus revealed no differences in those of germinants, wild-type or morphologically aberrant unilateral diploidizing strains of vegetative mycelium or in the clamp connection of the dikaryon. Simple septa of two general types prevailed in the common-A heterokaryon of S. commune.     

The liguleless2 gene of maize functions during the transition from the vegetative to the reproductive shoot apex

During a maize plant's (Zea mays) development, the shoot apical meristem (SAM) generates an apex that proceeds through different phases: juvenile vegetative, adult vegetative and reproductive. During each phase the structures produced are distinguishable from structures produced during the other phases. In this paper, we demonstrate that the LIGULELESS2 (LG2) function is required for an accurate vegetative to reproductive phase transition. The maize gene liguleless2 (lg2) has been shown to encode a basic-leucine zipper (bZIP) protein and to function in narrowing the region from which the ligule and auricle develop in a typical maize leaf. Here we show that lg2 mutant plants can have reduced long tassel branches, extra vegetative leaves and extra husk leaves when compared to wild-type siblings. This indicates a role for the lg2 gene in the vegetative to reproductive phase transition of the shoot apex. We also discuss a potential role for the lg2 gene in general phase transition processes.     

The rice FON1 gene controls vegetative and reproductive development by regulating shoot apical meristem size

Most plant organs develop from meristems. Rice FON1, which is an ortholog of Clv1, regulates stem cell proliferation and organ initiation. The point muta-tions, fon1-1 and fon1-2, disrupt meristem balance, resulting in alteration of floral organ numbers and the architecture of primary rachis branches. In this study, we identified two knockout alleles, fon1-3 and fon1-4, generated by T-DNA and Tos17 insertion, respectively. Unlike the previously isolated point mutants, the null mutants have alterations not only of the reproductive organs but also of vegetative tissues, producing fewer tillers and secondary rachis branches. The mutant plants are semi-dwarfs due to delayed leaf emergence, and leaf senescence is delayed. SEM analysis showed that the shoot apical meristems of fon1-3 mutants are enlarged. These results indicate that FON1 controls vegetative as well as reproductive development by regulating meristem size.     

Morphology, anatomy and reproduction of the eastern Mediterranean coralline Tenarea tortuosa and its relationship to members of the Lithophylloideae and Mastophoroideae (Rhodophyta, Corallinales)

One of the most characteristic organisms of the Mediterranean flora, the endemic coralline Tenarea tortuosa , has been collected along the Cretan coast and its vegetative and reproductive structures are reassessed. Gametophytes are described for the first time and are rare, observed only in one locality on the south coast. Reproductive structures suggest that the sexual life history is triphasic, but the present collections indicate unequal representation of males, carposporophytes and tetrasporophytes, and a dominance of bisporophytes suggesting that the alga is reproducing mainly asexually. The frond of Tenarea is formed by the coordinated growth of two elements (branches), growing back-to-back, each showing a simple Titanodema-type of organization; i.e. a monostromatic hypothallium of sinuous cells and a single layer of obliquely borne epithallial cells. Contiguous cells of the two opposite branches are not connected by secondary pit-plugs, and easily glide apart if decalcified. Natural splitting of the lamellae into its two branches may also occur, and single branches may fold regenerating the back-to-back organization. Analogous back-to-back growths occur in species of the distantly related genera Goniolithon (Lithophylloideae) and Leptophytum (Melobe-sioideae) and apparently represent cases of convergence. The affinities of Tenarea to related genera are discussed.     

Association of ergot alkaloids with conidiation in Aspergillus fumigatus

Ergot alkaloids are mycotoxins that affect the nervous and reproductive systems of exposed individuals through interactions with monoamine receptors. They have been studied more widely in ergot fungi and grass endophytes but also are found in Aspergillus fumigatus, an opportunistic human pathogen that reproduces and disseminates exclusively through conidia. The ergot alkaloids festucla-vine and fumigaclavines A, B and C are present in or on conidia of A. fumigatus. Cultures of the fungus that are free of conidia are difficult to obtain, obscuring comparisons of conidia versus vegetative hyphae as sources of the ergot alkaloids. To create conidiation-deficient strains of A. fumigatus we manipulated the bristle A gene (brlA), which controls vesicle formation or budding growth necessary for conidiation in Aspergillus spp. Disruption of brlA in A. fumigatus, via homologous recombination, resulted in a nonconidiating mutant that produced bristle-like structures instead of conidiophores and conidia. Moreover the disrupted strain failed to produce ergot alkaloids as verified by HPLC analyses. Complementation with a wild-type allele restored conidiation and ergot alkaloid production. These results suggest that ergot alkaloids are not produced within the vegetative mycelium of the fungus and are associated directly with conidiation.     

The Relationship between auxin transport and maize branching

Maize (Zea mays) plants make different types of vegetative or reproductive branches during development. Branches develop from axillary meristems produced on the flanks of the vegetative or inflorescence shoot apical meristem. Among these branches are the spikelets, short grass-specific structures, produced by determinate axillary spikelet-pair and spikelet meristems. We investigated the mechanism of branching in maize by making transgenic plants expressing a native expressed endogenous auxin efflux transporter (ZmPIN1a) fused to yellow fluorescent protein and a synthetic auxin-responsive promoter (DR5rev) driving red fluorescent protein. By imaging these plants, we found that all maize branching events during vegetative and reproductive development appear to be regulated by the creation of auxin response maxima through the activity of polar auxin transporters. We also found that the auxin transporter ZmPIN1a is functional, as it can rescue the polar auxin transport defects of the Arabidopsis (Arabidopsis thaliana) pin1-3 mutant. Based on this and on the groundbreaking analysis in Arabidopsis and other species, we conclude that branching mechanisms are conserved and can, in addition, explain the formation of axillary meristems (spikelet-pair and spikelet meristems) that are unique to grasses. We also found that BARREN STALK1 is required for the creation of auxin response maxima at the flanks of the inflorescence meristem, suggesting a role in the initiation of polar auxin transport for axillary meristem formation. Based on our results, we propose a general model for branching during maize inflorescence development.     

Axillary meristem development in Arabidopsis thaliana

Axillary shoot apical meristems initiate post-embryonically in the axils of leaves. Their developmental fate is a main determinant of the final plant body plan. In Arabidopsis, usually a single axillary meristem initiates in the leaf axil even though there is developmental potential for formation of multiple branches. While the wild-type plants rarely form multiple branches in the leaf axil, tfl1-2 plants regularly develop two or more branches in the axils of the rosette leaves. Axillary meristem formation in Arabidopsis occurs in two waves: an acropetal wave forms during plant vegetative development, and a basipetal wave forms during plant reproductive development. We report here the morphological and anatomical changes, and the STM expression pattern associated with the formation of axillary and accessory meristems during Arabidopsis vegetative development.     

Parasexually produced hybrids between female and male plants of Griffithsia tenuis C. Agardh,a red alga

Somatic cell fusion between vegetative cells of a male and a female isolate of Griffithsia tenuis, a marine red alga, has been obtained. Hybrid cells have been isolated and they have regenerated new plants. Almost all these hybrid plants made reproductive structures. In nearly half these cases the first 3–10 cells of the hybrid filament produced reproductive structures chracteristic of the tetrasporic (diploid) phase rather than the sexual (haploid) phase of the life cycle of this alga. However as these filaments continued to grow, cells further along the filament began to produce sexual, either female or male, reproductive structures. The observations suggest that the production of tetrasporangial branches does not require the fusion of male and female nucleic; rather, male and female nucleic remaining separate, act in concert to produce these structures, and in subsequent cell divisions the nuclei of one sex may be left behind allowing the nuclei of the remaining sex to direct the production of sexual branches.     

Distribution of plant‐dwelling spiders: Inflorescences versus vegetative branches

Abstract The influence of the architecture of vegetative branches on the distribution of plant‐dwelling spiders has been intensively studied, and the effects on the aggregation of individuals in several spider species on plants include variation in prey abundance, availability of predator‐free refuges and smoother microclimate conditions. The emergence of inflorescences at the reproductive time of the plants changes branch architecture, and could provide higher prey abundance for the spiders. The distribution of spiders between inflorescences and vegetative branches was compared on four widespread plant species in a Brazilian savannah‐like system. Inflorescences attracted more spiders than vegetative branches for all plant species sampled. The influence of branch type (inflorescence and vegetative) on spider distribution was also evaluated by monitoring branches of Baccharis dracunculifolia DC. in vegetative and flowering periods for 1 year, and through a field experiment carried out during the same period where artificial inflorescences were available for spider colonization. Artificial inflorescences attached to B. dracunculifolia branches attracted more spiders than non‐manipulated vegetative branches for most of the year. However, this pattern differed among spider guilds. Foliage‐runners and stalkers occurred preferentially on artificial inflorescences relative to control branches. The frequencies of ambushers and web‐builders were not significantly different between treatment and control branches. However, most ambush spiders (65%) occurred only during the flowering period of B. dracunculifolia, suggesting that this guild was influenced only by natural inflorescences. The experimental treatment also influenced the size distribution of spiders: larger spiders were more abundant on artificial inflorescences than on vegetative branches. The hypothesis that habitat architecture can influence the spider assemblage was supported. In addition, our observational and experimental data strongly suggest that inflorescences can be a higher quality microhabitat than non‐reproductive branches for most plant‐dwelling spiders.