Promotion of hyphal growth in Ustilago violacea by host factors from Silene alba

Ustilago violacea specifically parasitizes susceptible members of the Caryophyllaceae. We isolated water-soluble compounds from leaves of Silene alba which promoted hyphal development in the dimorphic pathogen. We also isolated hyphal growth promoting -tocopherol from S. alba. The water-soluble activity, which we term hyphal growth factor, or HGF, separated into four bands with gel filtration chromatography and represented over 40% of the total hyphal growth promoting activity isolated from S. alba. The water-soluble HGF activity may be host-specific and may function as a determinant of the host-parasite specificity between U. violacea and caryophyllaceous host plants.Abbreviations HGF Hyphal growth factor - BHT butylated hydroxytoluene     

Detection and identification of ferricrocin produced by ectendomycorrhizal fungi in the genusWilcoxina

The ectendomycorrhizal fungiWilcoxina mikolae isolates CSY-14 and RMD-947 andW. rehmii isolate CSY-85 were grown in pure culture under iron-limiting conditions. All three isolates tested positive for siderophore formation using both the ferric perchlorate assay and a sensitive HPLC iron-binding assay. A peptide siderophore was isolated from the culture medium by HPLC and shown to contain the amino acids serine, glycine and ornithine in a 1:2:3 ratio. This siderophore was identified as ferricrocin on the basis of electrospray mass spectroscopy and its co-chromatography in two different HPLC systems with ferricrocin isolated fromAspergillus fumigatus. Ferricrocin was the only siderophore isolated from theseWilcoxina cultures. This is the first report of siderophore formation by ectendomycorrhizal fungi.     

Biology and life cycle ofAtelocauda koae,an unusual demicyclic rust

Atelocauda koae, a rust of the native HawaiianAcacia koa, is considered as a demicyclic species, having spermogonial, aecial, and telial states, but is unusual in production of aeciospores simultaneously with teliospores rather than consecutively. Host inoculation with spores of each state separately confirmed that the life cycle was perpetuated by the telial state, but the aeciospores, while capable of germination and stomal penetration, did not produce detectable infection. This rust therefore behaves as a microcyclic species, and appears to be in evolutionary transition toward this reduced state. Teliospores produced vestigial, permanently attached basidiosopores which germinated to produce infective hyphae. The hyphae entered the host either through stomata or penetrated the epidermis directly, with the latter method being more common. Unusual nuclear associated with teliospore germination, in which meiosis occurs in more than one diploid nucleus was observed, in confirmation of an earlier study.     

Using 137Cs technique to quantify soil conservation capacities of different ecosystems in Wolong Natural Reserve, southwestern China

Reliable information about soil conservation capacities of different natural ecosystems is an important reference for the design of targeted erosion and sediment control strategies. The objective of this paper is to quantify the soil conservation capacities of different natural ecosystems that can represent different climatic zones. The 137Cs technique has been used to estimate soil redistribution rates in different natural ecosystems over the past 40 years in Wolong Nature Reserve. The reserve, transiting from the Chengdu plain to the Qinghai-Tibet plateau, maintains rich ecosystems from subtropical to frigid.The net soil erosion rates of 5 selected ecosystems that represent a warm coniferous-broadleaf-mixed forest, a cold-resistant deciduous taiga forest, a cold-resistant shrub, an evergreen cold-resistant taiga servation capacities are reversed in order. The reference inventories for 137Cs in different ecosystems velop effective erosion and sediment strategies in areas with similar climates should consider natural ecosystem types.     

Associations between primates and other mammals in a central Amazonian forest landscape

Little information exists on mixed-species groups between primates and other mammals in Neotropical forests. In this paper, we describe three such associations observed during an extensive large-vertebrate survey in central Amazonia, Brazil. Mixed-species groups between a primate species and another mammal were observed on seven occasions between squirrel monkeys (Saimiri cf. ustus) and either South American coatis (Nasua nasua) or tayras (Eira barbara) and between brown capuchins (Cebus apella) and coatis. All associations were restricted to floodplain forest during its dry stage. We suggest that the associations involving the coatis are connected to foraging and vigilance but may be induced by a common alternative food resource at a time of food shortage.     

Development of a new vitrification solution, VSL, and its application to the cryopreservation of gentian axillary buds

Vitrification methods are convenient for cryopreserving plant specimens, as the specimens are plunged directly into liquid nitrogen (LN) from ambient temperatures. However, tissues and species with poor survival are still not uncommon. The development of vitrification solutions with high survival that cover a range of materials is important. We attempted to develop new vitrification solutions using bromegrass cells and found that VSL, comprising 20% (w/v) glycerol, 30% (w/v) ethylene glycol, 5% (w/v) sucrose, 10% (w/v) DMSO and 10 mM CaCl₂, gave the highest survival following cryopreservation, as determined by fluorescein diacetate staining. However, the cryopreserved cells showed little regrowth, for unknown reasons. To check its applicability, VSL was used to cryopreserve gentian axillary buds and the performance was compared with those of conventional vitrification solutions. Excised gentian stem segments with axillary buds (shoot apices) were two-step precultured with sucrose to induce osmotic tolerance prior to cryopreservation. Gentian axillary buds cryopreserved using VSL following the appropriate preculturing approach exhibited 78% survival (determined by the regrowth capacity), which was comparable to PVS2 and PVS1 and far better than PVS3. VSL had a wider optimal incubation time (20–45 min) than PVS2 and was more suitable for cryopreserving gentian buds. The optimal duration of the first step of the preculture was 7–11 days, and preculturing with sucrose and glucose gave a much higher survival than fructose and maltose. VSL was able to vitrify during cooling to LN temperatures, as glass transition and devitrification points were detected in the warming profiles from differential scanning calorimetry. VSL and its derivative, VSL+, seem to have the potential to be good alternatives to PVS2 for the cryopreservation of some materials, as exemplified by gentian buds. Mitsuteru Suzuki, Pramod Tandon and Masaya Ishikawa contributed equally to the work.     

Bioenergetic consequences of glucoamylase production in carbon-limited chemostat cultures ofAspergillus niger

Aspergillus niger has been grown in glucose- and maltose-limited continuous cultures to determine the bioenergetic consequences of the production of the extracellular enzyme glucoamylase. Growth yields (g biomass per mol substrate) were high, indicating that growth was very efficient and protein production for biomass was not exceedingly energy consuming. It has been found that the energy costs for the production of this extracellular enzyme is very high. Depending on the efficiency of energy conservation the glucoamylase protein yield on ATP is between 1.3 and 2.6 g protein per mol ATP, which is equal or less than 10% of the theoretical maximum of 25.5. These high energy costs most probably have to be invested in the process of excretion. A comparison between an industrial over-producing strain and the wild typeAspergillus niger showed that this over-producing strain most probably is a regulatory mutant. Two regions of specific growth rates could be determined (one at specific growth rates lower and one at specific growth rates higher than 0.1 h^-1), which are characterized by differences in mycelium morphology and a significant deviation from linearity in the linear equation for substrate utilization. Analysis of the region of specific growth rates higher than 0.1 h^-1 yielded maintenance requirements of virtual zero. It has been concluded that for a good analysis of the growth behaviour of filamentour fungi the linear equation for substrate utilization is not suitable, since it contains no term for the process of differentiation.     

Nucleotide sequence and style-specific expression of a novel proline-rich protein gene from Nicotiana alata

cDNA clones encoding a novel proline-rich protein (NaPRP4) have been isolated from a Nicotiana alata stylar cDNA library. The N-terminal part of the derived protein is highly rich in proline (32.2%) and contains several repeats such as Lys-Pro-Pro (7 times) and Pro-Thr-Lys-Pro-Pro-Thr-Tyr-Ser-Pro-Ser-Lys-Pro-Pro (twice); the C-terminal part, on the other hand, has a lower proline content (9.9%) and contains two potential N-glycosylation sites and all the six cysteine residues. Northern blot and in situ hybridisation analyses indicate that expression of the NaPRP4 gene is restricted to cells of the transmitting tract of the style.     

Genetic variation in nine Shorea species (Dipterocarpaceae) in Indonesia revealed by AFLPs

Shorea is the largest and most important genus of the Dipterocarpaceae. The genetic diversity and structure of nine Shorea species from two different locations, namely Nanjak Makmur in Sumatra and Sumalindo in Borneo, were evaluated using amplified fragment length polymorphism (AFLP) markers. A total of 274 trees were investigated at 85 polymorphic AFLP loci. Levels of genetic diversity of these species ranged from  = 0.100 for S. acuminata to  = 0.165 for S. blumutensis. The population of rare species S. blumutensis possessed the highest genetic diversity suggesting that geographically restricted species can have levels of genetic variation comparable to closely related widespread common congeners. Analyses of molecular variance revealed that the genetic variation was mainly found among species in both locations (57.7% in Sumatra; 56.3% in Borneo). The unweighted pairgroup method using arithmetic averages dendrogram of all samples revealed an almost complete separation of species. Thus, AFLP markers proved appropriate for phylogenetic studies of Shorea species. Specific markers have been detected showing high-frequency differences among species and between regions within species. Sequence information of these markers can be used to develop specific polymerase chain reaction markers for wood identification. The possibility of interspecific hybridization was discussed.