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Behaviour of the hyphae of Laccaria laccata in the presence of Trichoderma harzianum in vitro.

The growth rate and the behaviour of Laccaria laccata and Trichoderma harzianum hyphae in co-culture and in the rhizosphere of 3-month-old Pinus sylvestris seedlings grown in vitro were investigated. In the interaction zone, hyphae of L. laccata became more pigmented and formed short branches growing towards the hyphae of the saprobic fungus, coiled around them and penetrated sporadically. Vacuolated hyphae of T. harzianum showed protoplasm granulation and breaks in walls followed by release of protoplasts. In the rhizosphere, the mantle hyphae of L. laccata showed a tendency to surround conidia of T. harzianum. No obvious penetration of the conidial walls by the hyphae of the mycorrhizal fungus was observed by scanning electron microscopy. Instead, in rare cases, the hyphae of L. laccata showed marked wrinkles, and a partial degradation of a mucilaginous material covering the mantle appeared to occur.

Agaricales↗

Mycorrhizal synthesis of Lactarius indigo (Schw.) Fr. with five Neotropical pine species.

This paper describes for the first time the ectomycorrhiza synthesized between two Guatemalan strains of Lactarius indigo (Schw.) Fr. and the Neotropical species Pinus ayacahuite var. ayacahuite Ehren, P. hartwegii Lindl., P. oocarpa Schiede ex Schltdl. var oocarpa, P. pseudostrobus Lindl. and P. rudis Endl. The synthesis was carried out in a controlled growth chamber using plastic containers with peat moss-vermiculite substrate and mycelial inoculum. Mycorrhiza were obtained 25 days after inoculation. A description of the morphology, appearance and structure of mantle and Hartig net is given for each combination. Mycorrhiza were saffron to cinnamon greenish with age, with a net of saffron laticifers visible through outer mantle; orange latex secreted when injured. Cystidia-like emanating hyphae were observed on the mantle surface of young mycorrhiza. Plectenchymatous mantle with abundant interhyphal gelatinous material.

Agaricales↗

Fungal communities in mycorrhizal roots of conifer seedlings in forest nurseries under different cultivation systems, assessed by morphotyping, direct sequencing and mycelial isolation.

Fungi colonising root tips of Pinus sylvestris and Picea abies grown under four different seedling cultivation systems were assessed by morphotyping, direct sequencing and isolation methods. Roots were morphotyped using two approaches: (1) 10% of the whole root system from 30 seedlings of each species and (2) 20 randomly selected tips per plant from 300 seedlings of each species. The first approach yielded 15 morphotypes, the second yielded 27, including 18 new morphotypes. The overall community consisted of 33 morphotypes. The level of mycorrhizal colonisation of roots determined by each approach was about 50%. The cultivation system had a marked effect on the level of mycorrhizal colonisation. In pine, the highest level of colonisation (48%) was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls (71%). Direct internal transcribed spacer ribosomal DNA sequencing and isolation detected a total of 93 fungal taxa, including 27 mycorrhizal. A total of 71 (76.3%) fungi were identified at least to a genus level. The overlap between the two methods was low. Only 13 (13.9%) of taxa were both sequenced and isolated, 47 (50.5%) were detected exclusively by sequencing and 33 (35.5%) exclusively by isolation. All isolated mycorrhizal fungi were also detected by direct sequencing. Characteristic mycorrhizas were Phialophora finlandia, Amphinema byssoides, Rhizopogon rubescens, Suillus luteus and Thelephora terrestris. There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems.

Ascomycota↗

Successful inoculation of mature pine with Tricholoma matsutake.

Our finding demonstrates, for the first time, that the roots of mature pine trees can be successfully inoculated with a symbiotic ectomycorrhizal fungus, the valuable matsutake mushroom. Long root segments (ca. 5-10 mm in diameter, ca. 50 cm in length) of 50-year-old Pinus densiflora trees were excavated, washed, auxin-treated (2-5 mg indole butyric acid, IBA, per root) and incubated in moist Spagnum moss. Twelve months later, short roots were regenerated, of which approximately 90% were free of mycorrhizae. Mycorrhiza-free short roots were inoculated with mycelial pieces of Tricholoma matsutake and incubated further in a sterilized substrate. Four-and-a-half months later, roots putatively colonized by Matsutake were sampled near the inoculation points. A T. matsutake-specific ITS-rDNA fragment was amplified by nested PCR from approximately 80% of the root samples analyzed, whereas approximately 66% of the root samples processed for staining with Chlorazol black E displayed characteristic T. matsutake Hartig net structures. These results confirm the symbiotic infection of mature P. densiflora roots by matsutake.

Agaricales↗

Genetic mosaics in the massive persisting rhizosphere colony "shiro" of the ectomycorrhizal basidiomycete Tricholoma matsutake.

The ectomycorrhizal basidiomycete Tricholoma matsutake produces commercially valuable fruit bodies "matsutake" on a massive persisting rhizosphere aggregate of mycelia and mycorrhizas called "shiro." Using inter-retrotransposon amplified polymorphism analysis, we attempted to explore the potential diversity within the population of T. matsutake isolated from small Pinus densiflora woodlands located in various parts of Japan. In general, random phylogenetic relationship was noted among T. matsutake tested. The population from each limited sampling area was highly heterogeneous. Even some isolates from fruit bodies produced in the same shiro and those from spores in the same fruit bodies were found to be genetically diverse, indicating the occurrence of genetic mosaics in shiro. In a mosaic shiro, heterologous genets produced their fruit bodies concurrently. Data suggested that the dispersal of spores through sexual reproduction may have been more prevalent than generally accepted in T. matsutake to bring mosaicism and coordination of heterologous genets within the shiro. Implementation of management taking such diversity into consideration is urgently needed for the restoration of devastated matsutake fields in Japan. Exploration of individual clones in mosaic fungal resources that promote colonization and fruit body production is necessary for it.

Basidiomycota↗

Phosphorus source alters host plant response to ectomycorrhizal diversity.

We examined the influence of phosphorus source and availability on host plant (Pinus rigida) response to ectomycorrhizal diversity under contrasting P conditions. An ectomycorrhizal richness gradient was established with equimolar P supplied as either inorganic phosphate or organic inositol hexaphosphate. We measured growth and N and P uptake of individual P. rigida seedlings inoculated with one, two, or four species of ectomycorrhizal fungi simultaneously and without mycorrhizas in axenic culture. Whereas colonization of P. rigida by individual species of ectomycorrhizal fungi decreased with increasing fungal richness, colonization of all species combined increased. Plant biomass and N content increased across the ectomycorrhizal richness gradient in the organic but not the inorganic P treatment. Plants grown under organic P conditions had higher N concentration than those grown under inorganic P conditions, but there was no effect of richness. Phosphorus content of plants grown in the organic P treatment increased with increasing ectomycorrhizal richness, but there was no response in the inorganic P treatment. Phosphorus concentration was higher in plants grown at the four-species richness level in the organic P treatment, but there was no effect of diversity under inorganic P conditions. Overall, few ectomycorrhizal composition effects were found on plant growth or nutrient status. Phosphatase activities of individual ectomycorrhizal fungi differed under organic P conditions, but there was no difference in total root system phosphatase expression between the inorganic or organic P treatments or across richness levels. Our results provide evidence that plant response to ectomycorrhizal diversity is dependent on the source and availability of P.

Biodiversity↗

Molecular characterization and evaluation of mycorrhizal capacity of Suillus isolates from central Spain for the selection of fungal inoculants.

Suillus fungal specimens of pine forests from a Mediterranean area of central Spain (Madrid region) were studied based on molecular and physiological analysis of sporocarps to obtain fungal native inocula to produce mycorrhizal Pinus halepensis Miller in nursery. Variation within the internal transcribed spacer (ITS) region of the ribosomal RNA genes of Suillus was examined by restriction fragment length polymorphism (RFLP) and direct sequencing of polymerase chain reaction products. Ribosomal DNA (rDNA) spacers were amplified from pure cultures obtained from fruit bodies of a range of Suillus species: Suillus bellinii (Inzenga) Watling, Suillus bovinus (Pers.) Kuntze, Suillus collinitus (Fr.) Kuntze, Suillus granulatus (L.) Snell, Suillus mediterraneensis (Jacquet. & Blum) Redeuil, Suillus luteus L. (Gray), and Suillus variegatus (Sw.) Kuntze. Interspecific variation in the length and number of restriction sites of the amplified ITS region was observed. This variation was confirmed by sequencing, which allowed us to identify some isolates. This is the first time that the ITS sequence of S. mediterraneensis is completely described. No intraspecific rDNA variation was observed within isolates of S. collinitus, S. mediterraneensis, and S. luteus. The phylogenetic analysis established the close relationship among these Mediterranean fungal species. As a further step to characterize the different isolates and to understand the relation between genetic and functional diversity, some physiological variables were evaluated. Intraspecific variation in axenic fungal growth and in mycorrhizal capacities was detected, especially within S. collinitus isolates. The fungal isolates stimulated the growth of P. halepensis in different rates. These studies indicated that ITS analysis, in conjunction with mycorrhizal tests, provides suitable combined tools for the analysis of Suillus spp. in a small geographic area for selecting isolates with final afforestation purposes.

Agaricales↗

Mycorrhization helper bacteria: a case of specificity for altering ectomycorrhiza architecture but not ectomycorrhiza formation.

Mycorrhization helper bacteria (MHB), isolated from phylogenetically distinct ectomycorrhizal symbioses involving Lactarius rufus, Laccaria bicolor or Suillus luteus, were tested for fungus specificity to enhance L. rufus-Pinus sylvestris or L. bicolor-P. sylvestris mycorrhiza formation. As MHB isolated from the L. rufus and S. luteus mycorrhiza were originally characterised using a microcosm system, we assessed their ability to enhance mycorrhiza formation in a glasshouse system in order to determine the extent to which MHB are system-specific. Paenibacillus sp. EJP73, an MHB for L. rufus in the microcosm, significantly enhanced L. bicolor mycorrhiza formation in the glasshouse, demonstrating that the MHB effect of this bacterium is neither fungus-specific nor limited to the original experimental system. Although the five MHB strains studied were unable to significantly enhance L. rufus mycorrhiza formation, two of them did have a significant effect on dichotomous short root branching by L. rufus. The effect was specific to Paenibacillus sp. EJP73 and Burkholderia sp. EJP67, the two strains isolated from L. rufus mycorrhiza, and was not associated with auxin production. Altered mycorrhiza architecture rather than absolute number of mycorrhizal roots may be an important previously overlooked parameter for defining MHB effects.

Bacteria↗

Trichloroacetic acid as additional factor contributing to desertification in southern Africa.

There has been an accelerated expansion of deserts in the past five decades. Recent data reveal that the atmogenically formed organic compound, trichloroacetic acid (TCA) is a contributory factor in addition to anthropogenic and natural sources. The aim of this study was to use TCA as an indicator for the possible occurrence of C2-chloroacetic acids; to assess the burden on the vegetation by using pine needles as a bio-indicator system and to deliberate on the possible role of TCA in the dynamics of the vegetation in southern Africa. Field experiments conducted on pine trees and on C3 and C4 crop plants under controlled laboratory conditions, have revealed that plants could be influenced positively or negatively by TCA. To obtain an integrated assessment of the pollution emission over a time span of at least one year, two-year-old pine needles of different Pinus species were used as a bio-indicator for TCA pollution at different measuring sites. The data of our investigation clearly indicate that areas exist in South Africa where the vegetation is burdened by ecotoxicologically relevant TCA contents comparable to those in central Europe and southern Russia where TCA was shown to play a role in the destabilisation of the steppe vegetation.

Air Pollutants↗

The chemistry of soils, rocks and plant bioindicators in three ecosystems of the Holy Cross Mountains, Poland.

In June of 2000, biogeochemical study was carried out in three ecosystems of the Holy Cross Mountains (south-central Poland). This paper presents element concentrations and stable sulfur (and in one site lead) isotope ratios in rocks, detailed soil profiles, and plant bioindicators including epiphytic lichen Hypogymnia physodes (L.) Nyl., mosses (Pleurozium schreberi (Brid.) Mitt., Hypnum cupressiforme Hedw. S. Str., Hylocomium splendens (Hedw.) B.S.G.), Scots pine (Pinus sylvestris L.), common birch (Betula pendula Roth.), aspen (Populus tremula L.) and English oak (Quercus robur L.). Chemical analyses were performed with ICP-AES and AAS methods in the accredited laboratory (Central Chemical Laboratory of the Polish Geological Institute in Warsaw). The principal objective of this study was to compare the chemical composition of rocks, soils and selected plant bioindicators between investigation sites using the same methods of sample collection, preparation and analyses. The results of this study have shown that there is a high variability in concentrations of elements in plant bioindicators from the same sites that can not be explained only by soil properties or anthropogenic influence. This conclusion indicates that for biomonitoring purposes (especially with vascular plants) we cannot neglect individual features of the species examined.

Betula↗

Relation between individual tree mortality and tree characteristics in a polluted and non-polluted environment.

UNLABELLED: Data on individual tree mortality in relatively healthy (Berezinskiy biosphere reserve) and damaged (surroundings of the mineral fertilizer plant 'Achema') even-aged Scots pine (Pinus sylvestris L.) stands are presented. Tree size and competition intensity were found to be the most significant predictors of individual tree mortality in all age groups of the relatively healthy Scots pine stands, however, an essential reduction in the closeness of relations between the tree mortality rate and these variables was determined with the aging of stands. An exponential decrease in tree mortality probability with an increase of tree size is characteristic for trees suffering different competition intensity, however, this decrease becomes much more pronounced as the competition pressure increases. The relations of different tree and stand variables with tree mortality probability have been found to become much weaker in the polluted environment. An exponential increase in tree mortality probability with an increase of crown defoliation was characteristic of damaged Scots pine stands, however, the rate of the increase was different in different age and diameter classes. The impact of crown defoliation on tree mortality rate increased with the aging of stands. At the same defoliation level, individual tree mortality probability was much higher for smaller suppressed trees, however, a relative increase in tree mortality probability along with an increase of crown defoliation was more pronounced for dominant trees. CONCLUSION: a higher mortality of damaged (defoliated) trees should be considered while assessing losses in forest productivity in a polluted environment.

Air Pollutants↗

Identification of sex pheromone components of a New Zealand geometrid moth, the common forest looper Pseudocoremia suavis, reveals a possible species complex.

Gas chromatography-electroantennographic detection analysis of sex pheromone gland extracts of the common forest looper Pseudocoremia suavis (Lepidoptera: Geometridae), a polyphagous defoliator of introduced Pinaceae and many New Zealand trees, revealed four compounds that elicited antennal responses. The two major active compounds (6Z)-cis-9,10-epoxynonadec-6-ene and (3Z,6Z)-cis-9,10-epoxynonadeca-3,6-diene were identified by comparison with known standards. Of the two minor active compounds, one was tentatively identified as (3Z,6Z)-cis-9,10-epoxyhenicosa-3,6-diene, whereas the other could not be identified because of insufficient amounts in extracts. (6Z)-cis-9,10-Epoxynonadec-6-ene, (3Z,6Z)-cis-9,10-epoxynonadeca-3,6-diene, and (3Z,6Z)-cis-9,10-epoxyhenicosa-3,6-diene were present in P. suavis gland extracts from Eyrewell Forest, a Pinus radiata plantation in the South Island of New Zealand, in a ratio of 35:65:5, respectively. Trapping trials in Eyrewell Forest established that (6Z)-cis-9,10-epoxynonadec-6-ene attracted male P. suavis. However, addition of (3Z,6Z)-cis-9,10-epoxyhenicosa-3,6-diene to the lure at <10% of (6Z)-cis-9,10-epoxynonadec-6-ene reduced capture of male moths, suggesting that one of its enantiomers was acting as a behavioral antagonist. During January-March of 2005, a blend trial involving single, binary, and ternary mixtures of the three components at Eyrewell Forest and at three other sites (two in the South Island and one in the North Island) revealed the existence of a second taxon of P. suavis at the three additional sites that was attracted to lures containing (3Z,6Z)-cis-9,10-epoxynonadeca-3,6-diene, either singly or in binary and ternary mixtures with (6Z)-cis-9,10-epoxynonadec-6-ene and (3Z,6Z)-cis-9,10-epoxyhenicosa-3,6-diene. This second taxon was not attracted to lures loaded solely with (6Z)-cis-9,10-epoxynonadec-6-ene.

Animals↗

Effect of intraspecific competition and substrate type on terpene emissions from some Mediterranean plant species.

Competition is an important factor that has been extensively reported in the Mediterranean area. There is evidence that leaf terpene accumulation may vary between plants growing on calcareous and siliceous soils. In the present study, leaf terpene emissions from potted seedlings of Pinus halepensis, Cistus albidus, and Quercus coccifera, growing under natural environmental conditions on calcareous and siliceous substrates, were studied by using a bag enclosure method. In both substrates, seedlings were potted alone and in intraspecific competition, to examine the effect of substrate type and that of intraspecific competition on terpene emissions. The results showed that competition favored: (i) overall monoterpene and sesquiterpene emissions from Q. coccifera; (ii) overall monoterpene emissions from P. halepensis; (iii) overall sesquiterpene emissions from C. albidus. Substrate type affected terpene emissions to a limited extent and in a species-specific way. Whereas for Q. coccifera, the overall monoterpene emissions and that of Allo-aromadendrene were favored on siliceous substrate, no significant changes were found in emissions from P. halepensis. Only the release of AR-curcumene from C. albidus was higher on siliceous substrate. We also found high variability in terpene emission composition from the study species, particularly for P. halepensis and Q. coccifera. These two species released both monoterpenes and sesquiterpenes, instead of monoterpenes only, as shown in previous studies.

Analysis of Variance↗

Large-scale statistical analysis of secondary xylem ESTs in pine.

A computational analysis of pine transcripts was conducted to contribute to the functional annotation of conifer sequences. A statistical analysis of expressed sequential tags(ESTs) belonging the 7732 contigs in the TIGR Pinus Gene Index (PGI1.0) identified 260 differentially represented gene sequences across six cDNA libraries from loblolly pine secondary xylem. Cluster analysis of this subset of contigs resulted in five groups representing genes preferentially represented in one of the xylem samples (compression wood, plannings, root xylem, latewood) and one group containing mostly genes simultaneously present in compression and side wood libraries. To complement the sequence annotation, 27 cDNA clones representing selected transcripts were completely sequenced. Several genes were identified that could represent putative markers for xylem from different organs, at different stages of development. Several sequences encoding regulatory proteins were over-represented in root xylem as opposed to the other xylem samples. Some of them belonged to known families of plant transcription factors, but two genes were previously uncharacterized in plants. One transcript was homologous to the gene encoding the Smad4 interacting factor, a key co-activator in TGFbeta (transforming growth factor) signalling in animals. Thus, the digital analysis of pine ESTs highlighted a putative gene function of potentially broad interest but that has yet to be investigated in plants. More generally, this study showed that the application of numerical approaches to EST databases should be helpful in establishing priorities among genes to consider for targeted functional studies. Thus, we illustrated the potential of extracting information from conifer sequences already accessible through well-structured public databases.

Amino Acid Sequence↗

Expressed sequence tags from loblolly pine embryos reveal similarities with angiosperm embryogenesis.

The process of embryogenesis in gymnosperms differs in significant ways from the more widely studied process in angiosperms. To further our understanding of embryogenesis in gymnosperms, we have generated Expressed Sequence Tags (ESTs) from four cDNA libraries constructed from un-normalized, normalized, and subtracted RNA populations of zygotic and somatic embryos of loblolly pine (Pinus taeda L.). A total of 68,721 ESTs were generated from 68,131 cDNA clones. Following clustering and assembly, these sequences collapsed into 5,274 contigs and 6,880 singleton sequences for a total of 12,154 non-redundant sequences. Searches of a non-identical amino acid database revealed a putative homolog for 9,189 sequences, leaving 2,965 sequences with no known function. More extensive searches of additional plant sequence data sets revealed a putative homolog for all but 1,388 (11.4%) of the sequences. Using gene ontologies, a known function could be assigned for 5,495 of the 12,154 total non-redundant sequences with 13,633 associations in total assigned. When compared to approximately 72,000 sequences in a collated P. taeda transcript assembly derived from >245,000 ESTs derived from root, xylem, stem, needles, pollen cone, and shoot ESTs, 3,458 (28.5%) of the non-redundant embryo sequences were unique and thereby provide a valuable addition to development of a complete loblolly pine transcriptome. To assess similarities between angiosperm and gymnosperm embryo development, we examined our EST collection for putative homologs of angiosperm genes implicated in embryogenesis. Out of 108 angiosperm embryogenesis-related genes, homologs were present for 83 of these genes suggesting that pine contains similar genes for embryogenesis and that our RNA sampling methods were successful. We also identified sequences from the pine embryo transcriptome that have no known function and may contribute to the programming of gene expression and embryo development.

Amino Acid Sequence↗

Selective increase in pinolenic acid (all-cis-5,9,12-18:3) in Korean pine nut oil by crystallization and its effect on LDL-receptor activity.

The aims of this study were to obtain concentrated pinolenic acid (5,9,12-18:3) from dietary Korean pine (Pinus koraiensis) nut oil by urea complexation and to investigate its cholesterol-lowering effect on the LDL-receptor activity of human hepatoma HepG2 cells. Pine nut oil was hydrolyzed to provide a low-pinolenic acid-containing FA extract (LPAFAE), followed by crystallization with different ratios of urea in ethanol (EtOH) or methanol (MeOH) as a solvent to produce a high-pinolenic acid-containing FA extract (HPAFAE). The profiles of HPAFAE obtained by urea complexation showed different FA compositions compared with LPAFAE. The long-chain saturated FA palmitic acid (16:0) and stearic acid (18:0) were decreased with urea/FA ratios (UFR) of 1:1 (UFR1), 2:1 (UFR2), and 3:1 (UFR3). Linoleic acid (9,12-18:2) was increased 1.3 times with UFR2 in EtOH, and linolenic acid (9,12,15-18:3) was increased 1.5 times with UFR3 in MeOH after crystallization. The crystallization with UFR3 in EtOH provided the highest concentration of pinolenic acid, which was elevated by 3.2-fold from 14.1 to 45.1%, whereas that of linoleic acid (9,12-18:2) was not changed, and that of oleic acid (9-18:1) was decreased 7.2-fold. Treatment of HepG2 cells with HPAFE resulted in significantly higher internalization of 3,3'-dioctadecylindocarbocyanine-LDL (47.0 +/- 0.15) as compared with treatment with LPAFAE (25.6 +/- 0.36) (P< 0.05). Thus, we demonstrate a method for the concentration of pinolenic acid and suggest that this concentrate may have LDL-lowering properties by enhancing hepatic LDL uptake.

Animals↗

Supplementation with a pine bark extract rich in polyphenols increases plasma antioxidant capacity and alters the plasma lipoprotein profile.

Pycnogenol (PYC), an extract of French maritime pine bark (Pinus pinaster), is a potent antioxidant with potential health benefits. Its bioavailabilty has previously been shown by urinary excretion studies of constituents and metabolites of PYC. The aim of this study was to test the effect of PYC supplementation on measures of oxidative stress and the lipid profile in humans. Twenty-five healthy subjects received PYC (150 mg/d) for 6 wk. Fasting blood was collected at baseline, after 3 and 6 wk of supplementation, and again after a 4-wk washout period. After 6 wk of supplementation with PYC, a significant increase in plasma polyphenol levels was detectable, which was reversed after the 4-wk washout phase. The antioxidant effect of PYC was demonstrated by a significant increase in oxygen radical absorbance capacity (ORAC) in plasma throughout the supplementation period (P < 0.05). The ORAC value returned to baseline after the 4-wk washout period. Moreover, in addition to its antioxidant effects, PYC significantly reduced LDL-cholesterol levels and increased HDL-cholesterol levels in plasma of two-thirds of the subjects. While the LDL changes reversed during washout, the HDL increase did not. There was no significant difference in LDL oxidizability or plasma lipid peroxides following PYC supplementation. Hence, following oral supplementation in humans, PYC significantly increases antioxidant capacity of plasma, as determined by ORAC, and exerts favorable effects on the lipid profile.

Administration, Oral↗

Biological investigations aboard biosatellite Cosmos-782.

The Cosmos-782 flight from 25 November to 15 December 1975, carried biological experiments designed to study the effects of weightlessness on insects and fish and on gravitropism and growth in several seed varieties. Investigations carried out on Drosophila melanogaster measured the frequency of recessive lethal mutations and the change in genetic distances in the sex chromosome. The study of Fundulus heteroclitus eggs and fry compared the effects of weightlessness and artificial gravity. Plants experiments studied spatial orientation of over and underground organs of Pinus silvestris and Crepis capillaris seeds. Other investigations used Phycomyces blakesleanus to compare spatial orientation and growth and development in weightlessness and artificial gravity.

Animals↗