Dynamics of the formation of secondary reproductive structures in Streptomyces zone-forming colonies as determined from confocal microscopy data

The dynamics of the formation of secondary structures on the colony surface, so-called air mycelium concentric rings, was investigated by confocal microscopy and using Streptomyces zone-forming colonies as a model of a self-organizing system. The occurrence of luminescence in reproductive structures was registered, whereas in vegetative structures it was almost completely absent. It was shown that secondary reproductive structures begin to form within the thickness of substratum mycelium as branches or as an apical extension of vegetative gyphes, followed by the formation of numerous secondary branches filled with protospores.     

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.     

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.     

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.     

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.     

Density-dependent reproductive and vegetative allocation in the aquatic plant Pistia stratiotes (Araceae)

Pistia stratiotes is an aquatic macrophyte that grows in temporary-ponds in the southern Pantanal, Brazil. It reproduces both sexually and asexually and is usually observed forming dense mats on the water surface, a condition favored by the plant's vegetative reproduction coupled with an ability for rapid growth. In this study we examined the effect of densely crowded conditions on the production of reproductive and vegetative structures. In addition, we verified whether there is a trade-off between clonal growth and investment in sexual reproductive structures, and whether there is an allocation pattern with plant size. Individual plant biomass and the number of the rosettes producing sexual reproductive structures and vegetative growth structures both increased with density. Increase in plant size resulted in increased proportional allocation to sexual reproductive structures and vegetative growth structures. Allocation of biomass to reproduction did not occur at the expense of clonal growth. Thus, the density response appears as a increase of rosettes producing sexual reproductive structures and vegetative growth structures. Therefore, long leaves and stolons may be adaptive under densely crowded conditions where competition for light is intense. An important aspect in the study of trade-offs is the size-dependency of the allocation patterns .Usually, larger plants produce more biomass. Therefore, larger plants can allocate more biomass to both vegetative and sexual reproduction than smaller plants and thus show a positive correlation between both traits rather than the expected negative one.     

Significance of the simultaneous growth of vegetative and reproductive organs in the prostrate annual Chamaesyce maculata (L.) Small (Euphorbiaceae)

To elucidate the significance of the simultaneous growth of vegetative and reproductive organs in the prostrate annual Chamaesyce maculata (L.) Small (Euphorbiaceae) from the standpoint of meristem allocation, we investigated plant architecture, meristem allocation, and the spatial and temporal patterns in vegetative growth and reproduction in the reproductive stage. The numbers of secondary and tertiary shoots successively increased by branching in the reproductive stage, and the sum of shoot length was greater in secondary shoots than in primary shoots. The specific shoot length (shoot length per shoot biomass) was greater in lateral shoots than in primary shoots, indicating efficient lateral shoot elongation. The internode length was shorter in secondary shoots than in primary shoots, increasing the number of nodes per shoot length in secondary shoots. Many nodes on a shoot generated two meristems, one of which committed to a flower and one to a lateral shoot. The number of reproductive meristems was greatest in tertiary shoots, and 96% of total reproductive meristems on shoots were generated in lateral shoots. On almost all nodes, the reproductive meristem developed into a flower, and 95–98% of the flowers produced a fruit. Therefore, vegetative growth by branching in the reproductive stage contributed to the increase in reproductive outputs. From the standpoint of meristem allocation, the simultaneous growth of vegetative and reproductive organs in prostrate plant species might be important for increasing the number of growth and reproductive meristems, resulting in the increase in reproductive outputs.     

Respiration and reproductive effort in Xanthium canadense

* BACKGROUND AND AIMS: The proportion of resources devoted to reproduction in the plant is called the reproductive effort (RE), which is most commonly expressed as the proportion of reproductive biomass to total plant biomass production (RE(W)). Reproductive yield is the outcome of photosynthates allocated to reproductive structures minus subsequent respiratory consumption for construction and maintenance of reproductive structures. Thus, RE(W) can differ from RE in terms of photosynthates allocated to reproductive structures (RE(P)). * METHODS: Dry mass growth and respiration of vegetative and reproductive organs were measured in Xanthium canadense and the amount of photosynthates and its partitioning to dry mass growth and respiratory consumption were determined. Differences between RE(W) and RE(P) were analysed in terms of growth and maintenance respiration. * KEY RESULTS: The fraction of allocated photosynthates that was consumed by respiration was smaller in the reproductive organ than in the vegetative organs. Consequently, RE(P) was smaller than RE(W). The smaller respiratory consumption in the reproductive organ resulted from its shorter period of existence and a seasonal decline in temperature, as well as a slower rate of maintenance respiration, although the fraction of photosynthates consumed by growth respiration was larger than in the vegetative organs. * CONCLUSIONS: Reproductive effort in terms of photosynthates (RE(P)) was smaller than that in terms of biomass (RE(W)). This difference resulted from respiratory consumption for maintenance, which was far smaller in the reproductive organ than in vegetative organs.     

Pattern of phosphoproteins during cell differentiation in Streptomyces collinus

Transition from vegetative cells to aerial mycelium and spores of Streptomyces collinus is accompanied by changes in the pattern of proteins phosphorylated. Preparation from spores exhibits lower phosphorylation activity than those of vegetative cells and aerial mycelium. Phosphorylation of proteins from aerial mycelium was markedly stimulated by the presence of Mn²⁺. Our data indicate that phosphorylation of proteins on Ser/Thr residues is involved in transition of vegetative cells to aerial mycelium.     

Developmental instability in gynodioecious Teucrium lusitanicum

Developmental instability was assessed in two geographical races of Teucrium lusitanicum using morphometric measures of vegetative and reproductive structures. T. lusitanicum is a gynodioecious species. Male sterile (female) individuals showed greater developmental instability at all sites. Plants located inland had higher developmental instability of vegetative characters and lower developmental instability of reproductive characters than coastal plants. These results support the contentions that (1) developmental instability is affected more by the disruption of co-adapted gene complexes than by lower heterozygosity, and (2) different habitat characteristics result in the differential response of vegetative and reproductive structures.     

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。     

Adenine and pyridine nucleotide levels during calcium-induced conidiation in Penicillium notatum

Concentrations of adenine and pyridine nucleotides and the associated charge values were examined in extracts of mycelium of Penicillium notatum during vegetative growth and reproductive development promoted by the addition of Ca²⁺ (10 mmol dm^-3). The significant increase in adenylate energy charge promoted by Ca²⁺ was due to a fall in intracellular AMP and a concomitant rise in ATP concentration. Intracellular concentrations of NADH and NAD fell within 1 h of the addition of Ca²⁺. The catabolic reduction charge was unchanged by Ca²⁺ whilst the anabolic reduction charge increased in Ca²⁺-induced mycelium due to lowered intracellular NADP concentration. Reduced concentration of NADPH in Ca²⁺-induced mycelium, relative to the vegetative controls, lowered the phosphorylated nucleotide fraction. The results are discussed in relation to metabolic economy during morphogenesis in P. notatum.     

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.     

沙棘根瘤内生菌的多型性

用透射电子显微镜观察了春、夏、秋、冬四个季节的沙棘根瘤,以及瘤瓣上、中、下三个部位。结果表明,不同季节,不同部位的瘤瓣内,根瘤内生菌有7种不同形态。即侵染菌丝体、繁殖菌丝体、营养菌丝体、春孢子及春孢子囊、泡囊,冬孢子及冬孢子囊和类菌体。在多年生珊瑚状的根瘤中,它们的世代交替是:春夏季以侵染菌丝、繁殖菌丝、营养菌丝、春孢子囊及春孢子、泡囊为主;秋冬季以衰退的营养菌丝、衰老泡囊、冬孢子囊和冬孢子、类菌体为主。冬孢子和类菌体是休眠体。     

高寒草甸矮蒿草种群繁殖对策的研究

繁殖对策是指生物对环境的生殖适应趋势 ,是资源或能量向生存、生长和生殖等活动中最适分配的结果 ,在不同的环境中具有其独特的表现形式。研究植物在不同环境中的繁殖对策可以反映出植物对环境的适应能力和在该生境中的生殖潜能。国内外学者对植物繁殖对策的研究已有不少报道[2 ,4 ,5,6] 。但对高寒草甸矮嵩草 (Ｋｏｂｒｅｓｉａｈｕｍｉｌｉｓ)种群繁殖对策的研究报道甚少。矮嵩草是青藏高原矮嵩草草甸的建群种 ,它具有草质柔软、营养丰富、热值含量较高等特点 ,是青藏高原重要的可更新草地资源。本研究对矮嵩草的繁殖对策进行了较全面、…     

Gibberellins promote vegetative phase change and reproductive maturity in maize.

Postembryonic shoot development in maize (Zea mays L.) is divided into a juvenile vegetative phase, an adult vegetative phase, and a reproductive phase that differ in the expression of many morphological traits. A reduction in the endogenous levels of bioactive gibberellins (GAs) conditioned by any one of the dwarf1, dwarf3, dwarf5, or another ear1 mutations in maize delays the transition from juvenile vegetative to adult vegetative development and from adult vegetative to reproductive development. Mutant plants cease producing juvenile traits (e.g. epicuticular wax) and begin producing adult traits (e.g. epidermal hairs) later than wild-type plants. They also cease producing leaves and begin producing reproductive structures later than wild-type plants. These mutations greatly enhance most aspects of the phenotype of Teopod1 and Teopod2, suggesting that GAs suppress part but not all of the Teopod phenotype. Application of GA3 to Teopod2 mutants and Teopod1, dwarf3 double mutants confirms this result. We conclude that GAs act in conjunction with several other factors to promote both vegetative and reproductive maturation but affect different developmental phases unequally. Furthermore, the GAs that regulate vegetative and reproductive maturation, like those responsible for stem elongation, are downstream of GA20 in the GA biosynthetic pathway.     

The evolution of new structures: clues from plant cytoskeletal genes.

How large numbers of genes were recruited simultaneously to build new organ structures is one of the greatest puzzles in evolutionary biology. Here, we present data suggesting that the vegetative and reproductive classes of actins and other cytoskeletal proteins arose concurrently with the macroevolutionary divergence of leaves and reproductive structures in the earliest land plants. That the cytoskeleton is essential for physically programming the development of organs and tissues is well established. Thus, we propose that this regulatory dichotomy represents an ancient landmark event in the global regulation of hundreds of higher-plant genes, an event that is linked to the macroevolution of plant vegetative and reproductive organs. The recent availability of sequence and expression data for large numbers of plant genes should make it possible to dissect this and other major macroevolutionary events.