Fungal endophytes from the leaves and twigs of Quercus ilex L. from England, Majorca and Switzerland.

Leaves of Quercus ilex taken from sites in England, Majorca and Switzerland have been studied to detect the influence of the geographic position of the host within and outside its native range on the composition of its endophytic fungal assemblages. Samples of stem tissue of Q. ilex collected from the Swiss trees were also studied to confirm tissue-specific differences. Sixty different fungal taxa were isolated, but only 28 were frequent. Of the total number of isolates from the leaves from the Swiss, British and Spanish sites 87%, 31% and 63%, respectively, were coelomycetes. Four species of Phomopsis, which includes Phyllosticta ilicina (=Phomopsis ilicina v. d. Aa, ined.), were the most frequent endophytes of leaves and were either absent or rare in the twig units. Two distinct kinds of sterile mycelia were common in twigs. Swiss and Spanish trees possessed fungal assemblages distinct from those present in Britain. Naturalized stands were distinguished from native stands by the presence of rather cosmopolitan and non-specific fungal taxa, rare or absent in the samples collected in the native stands. Samples derived from the native stands were colonized by more host-specific fungi. The relative frequency of two sterile mycelia in the Swiss and Spanish sites determined their separation. Phyllosticta (Phomopsis) ilicina, the most numerous leaf colonizer, was virtually absent from the bark and the xylem. The frequent occurrence of coelomycetes as endophytes of woody trees is briefly discussed.

Two undescribed hyphomycetes from Malaysia.

Raffaelea variabilis is described as a new species in culture from Lannea grandis. It is distinguished by turbinate to cuneiform conidia measuring 4-16 times 2.5-7.5 mum, and compared and contrasted with established species. Trichocladium lobatum is described as a new species in culture with 1-2 euseptate spherical conidia, 19-22 mum diam., ornamented with flabelliform, spathulate or petaloid lobes 7 mum long. It is compared and contrasted with established Trichocladium species and representatives of Chlamydomyces, Histoplasma, Mycogone, Sepedonium and Thermomyces.

Changes in two ribonuclease isozymes during rust infection of flax cotyledons.

A ribonuclease fraction previously purified from flax by gel filtration was further resolved into two components by hydroxyl apatite chromatography. These were homogeneous with respect to electrophoresis and isoelectric focusing. Both enzymes are of RNase I type but differ in substrate specificity, kinetic properties, pH response, and isoelectric point.The two RNase isozymes show consistent properties when extracted from variety Bison (susceptible) or variety Bombay (resistant) with or without infection with race 3 of flax rust. The relative amounts of these isozymes change markedly during infection. These observations provide an explanation for the apparent qualitative changes in RNase noted previously. Differences between susceptible and resistant reactions in the early stages of disease are discussed.

An integrated approach to Phoma systematics.

Physiological and morphological characters were recorded from 55 strains of 17 Phoma taxa and one Pyrenochaeta. The results were subjected to numerical analysis and UPGMA dendrograms produced. The full results were compared with TLC profiles of secondary metabolites. Seven distinct clusters were recovered from dendrograms based on full and partial character sets and the grouping of strains within each cluster discussed. The new combination Phoma sambuci-nigrae (Sacc.) Monte, Bridge & Sutton is proposed for P. herbarum f. sambuci-nigrae Sacc.

Pseudomicrodochium fusarioides sp. nov., isolated from human bronchial fluid.

A fungus from bronchial lavage fluid from a patient with pulmonary infiltration was isolated in pure culture and identified as a species of Pseudomicrodochium. It is compared and contrasted with other species of the genus, in particular P. suttonii, and formally described as P. fusarioides.

Vicilin-like seed storage proteins in the gymnosperm interior spruce (Picea glauca/engelmanii).

A seed storage protein cDNA was characterized from a library of interior spruce (Picea glauca/engelmanii complex) cotyledonary stage somatic embryos. The deduced amino acid sequence predicts a 448 amino acid (50 kDa) polypeptide with 28-38% identity with angiosperm vicilin-like 7S globulins. XXC/G codon usage is low (47%) relative to monocot angiosperms while pairwise comparisons show that spruce, monocot, and dicot vicilins are approximately equal in amino acid divergence. Although small by comparison, the spruce vicilin contains an N terminal hydrophilic region characteristic of angiosperm 'large' vicilins. Genomic Southern blotting predicts that the cDNA is encoded by a gene family.

Relationships between gas exchange adaptation of Sitka x interior spruce genotypes and ribosomal DNA markers.

Adaptive physiological changes were investigated in seven populations of Sitka (Picea sitchensis (Bong.) Carr.) x interior spruce (P. glauca (Moench) Voss x P. engelmannii Parry ex Engelm.) spanning the Nass-Skeena transition zone in British Columbia, Canada. Each population was represented by an Si rDNA index that was calculated from the relative optical densities on a gel autoradiogram of five ribosomal DNA bands characteristic of Sitka spruce and interior spruce. This index estimates the proportion of the genome contributed by interior spruce. Physiological adaptations were assessed by gas exchange parameters measured under both well-watered and drought conditions. Under well-watered conditions, Sitka spruce populations had higher maximal photosynthesis at saturating light and ambient CO(2), higher quantum yield at the light compensation point, and higher dark respiration than interior spruce populations. Sitka spruce populations also reached maximal photosynthesis at lower photosynthetically active radiation and higher CO(2) concentrations, and had higher stomatal densities that resulted in lower stomatal limitations to photosynthesis than interior spruce populations. In contrast, interior spruce populations exhibited greater drought tolerance than Sitka spruce populations. Their gas exchange rates declined at a slower rate in response to drought. They maintained higher gas exchange rates in response to moderate to severe drought (predawn plant water potentials = -1.5 MPa), and their photosynthetic rates recovered faster when they were rewatered after exposure to drought. Comparison of the seven populations indicated that physiological parameters were significantly related to the Si rDNA index. An increase in Si rDNA index was associated with proportional changes in physiological measurements, suggesting that genetic interchange among species with contrasting ecological adaptations can enhance the environmental adaptation of natural populations.

Storage protein changes during zygotic embryogenesis in interior spruce.

The major storage proteins isolated from protein bodies of embryo tissues of interior spruce Picea glauca (Moench) Voss/Picea engelmanii Parry had apparent molecular weights of 41, 35, 33, 24 and 22 kD. Minor proteins of 30 and 27.5 kD were also observed. Based on their solubility characteristics, the 41 kD protein was identified as a water and buffer-soluble albumin, and the 35, 33, 24 and 22 kD proteins were characterized as buffer-insoluble, high salt-soluble globulins. Two-dimensional electrophoresis revealed each protein was composed of several isoelectric variants. Developmentally specific accumulation of storage proteins was observed during embryogenesis. The 41 kD protein only accumulated during the later stages of cotyledon maturation, whereas the other storage proteins began to accumulate during the early stages of embryo development. All storage proteins showed major accumulations during cotyledon maturation.

Isolation and expression of Rhizobium japonicum cloned DNA encoding an early soybean nodulation function.

A first visible step in the nodulation of legumes by Rhizobium spp. is the deformation and curling of root hairs. We have identified and cloned DNA sequences encoding this function from two strains of Rhizobium japonicum (USDA 122 and USDA 110) with a weakly homologous probe from Rhizobium meliloti. Root hair curling encoded by the cloned DNA fragments was examined on soybeans (Glycine soja ) after conjugative transfer of these sequences in broad-host-range vectors to various bacterial genera. Pseudomonas putida gave unambiguous expression of the root hair curling genes. This enabled us to identify the 8.7-kilobase EcoRI fragments encoding root hair curling from each strain. The phenotypes encoded by the plasmids pBS1 (derived from strain USDA 122) and pBS2 (derived from strain USDA 110) are distinct and represent a phenotype characteristic of their parent R. japonicum strains. Subclones of pBS1 and pBS2 were generated in single and multicopy vectors, and their expression was analyzed in P. putida. We established that a 4.2-kilobase internal Sa/I fragment of pBS1 and a 3.5-kilobase SstI -EcoRI fragment of pBS2 are sufficient to confer root hair curling on soybeans.

Relationship between nuclear DNA markers and physiological parameters in Sitka x interior spruce populations.

Eight populations of Sitka spruce (Picea sitchensis (Bong.) Carr.) and interior spruce (Picea glauca (Moench) Voss x Picea engelmannii Parry ex. Engelm.) seedlings were sampled from a zone of Sitka-interior spruce introgression in British Columbia, Canada. Restriction fragment length polymorphisms of the nuclear ribosomal RNA genes (rDNA) were used to define species-specific hybridization patterns for the Sitka spruce and interior spruce populations. Hybridization was estimated from an index based on the relative abundance of polymorphic rDNA combining bands for each population. Sitka x interior hybrid seedlings had an index value for the relative abundance of interior spruce rDNA (Si-rDNA) ranging from 0.07 (Lower Nass; the most westerly collected source) to 0.95 (Bulkley Valley low-elevation seed orchard). During shoot elongation, osmotic potential at saturation (Psi(sat)) and turgor loss point (Psi(tlp)) increased, whereas total turgor (Psi(PTotal)) decreased. After bud set in the summer and throughout the fall, Psi(sat) and Psi(tlp) decreased, whereas Psi(PTotal) increased. At all times of year, populations with a higher Si-rDNA index had lower Psi(tlp) and Psi(sat) and higher Psi(PTotal) than populations with a lower Si-rDNA index. During the fall, Sitka x interior hybrid seedlings exhibited a seasonal decline in the temperature causing 50% needle electrolyte leakage (LT(50)) and in the critical temperature indicating the initial point of freezing injury. Seedlings with a higher Si-rDNA index had lower LT(50) and critical temperature values indicating greater freezing tolerance in the fall. Throughout most of the year, seedling population Si-rDNA index was related to the degree of drought and freezing tolerance.

Characterization of immature embryos of interior spruce by SDS-PAGE and microscopy in relation to their competence for somatic embryogenesis.

SDS-PAGE analysis of total proteins from cotyledonary embryo explants reveals that their competence to form somatic embryos is limited to a specific stage of development prior to the accumulation of storage proteins. When protein profiles of embryo explants of different open pollinated families from the same collection date are compared, there is a close relationship between the absence of storage proteins and their ability to produce embryogenic callus. In addition, the appearance of storage proteins in embryos from subsequent collections is associated with their loss of competence. Light microscopy combined with staining for total protein demonstrates that competent immature embryos have cotyledons but do not contain protein bodies.

Inheritance of chloroplast and mitochondrial DNA in Picea and composition of hybrids from introgression zones.

The cloning of white spruce (Picea glauca) mitochondrial DNA homologous to the cytochrome oxidase II and ATPaseα genes of maize is described. These probes were used to define restriction fragment length polymorphisms which distinguish the white, Engelmann (P. engelmannii) and Sitka spruce (P. sitchensis) populations that occur in British Columbia. Analysis of progeny from crosses between the species revealed that mitochondrial DNA was maternally inherited in all cases (32 progeny from five independent crosses). The inheritance of chloroplast DNA was determined using a probe described previously; in this case, all progeny exhibited paternal inheritance (27 progeny from four crosses). Mitochondrial and chloroplast probes were used to test trees from zones of introgression between coastal (Sitka) and interior spruces (white and Engelmann). In most cases mitochondria and chloroplasts within individuals were contributed by different species. The data shows that there is a significant Sitka spruce component in trees east of the coastal watershed in British Columbia.