Screening ofPenicillium species for the production of extracellular glucose oxidase

During screening ofPenicillium species for extracellular glucose oxidase production, a strain ofPenicillium variabile (P16) was selected which released high activities of enzyme into the culture medium. Maximum activity (5.49 U/ml) was after 96 h cultivation in shake-flask culture.M. Petruccioli is with the Dipartimento di Biologia, Difesa e Biotecnologie Agro-Alimentari, University of Basilicata, Via N. Sauro 85, 1-85100 Potenza, Italy. M. Ceccarelli and F. Federici are with the Dipartimento Agrobiologia e Agrochimica, University of Tuscia, Via S.C. de Lellis, 1-01100 Viterbo, Italy.     

Phytohormone changes during storage root growth in Beta species

The physiological and morphological factors necessary for efficient accumulation of sucrose in sugar beet (Beta vulgaris L.) are considered in relation to potential uses of plant growth regulators to modify the anatomy of storage roots so as to increase sucrose content and yield. The percentage of sucrose in root fresh and dry matter is closely related to root structure. Sugar beet, mangold and chard are three sub-species of Beta vulgaris that differ considerably in their anatomy, assimilate partitioning, sucrose concentration and root dry matter yield. The concentrations of indole-3-acetic acid (IAA), abscisic acid (ABA) and cytokinins were measured during the growth of the storage root in each of these cultivars. Correlations were found between the phytohormone levels and the formation of secondary cambia and their subsequent cell division and expansion activity.     

Identification ofPenicillium sp. NR 6564 and taxonomic notes onP. janthinellum

The fungal strain NR 6564, which produces a new series of antifungal antibiotics, Ro 09-1470 and its congeners, was identified asPenicillium janthinellum from its cultural and morphological properties. The identification was confirmed by analysis of the ubiquinone system and DNA-DNA hybridization.     

Energy storage during reproductive diapause in the Drosophila melanogaster species group

Summary Temperate species of the Drosophila melanogaster group enter reproductive diapause for overwintering in response to short daylength. During the prediapause period they accumulate triacylglycerols, but not glycogen, as energy resources. The capacity for storing triacylglycerols differs between species, and appears to be closely correlated with diapause and cold-hardiness; cool-temperate species, such as those of the auraria species complex, which enter a deep diapause and are highly cold-hardy, accumulate larger quantities of triacylglycerols than warm-temperate species, such as D. rufa and D. lutescens, which enter a weak diapause and are less cold-hardy. Among the cool-temperate spcies, D. subauraria occurs at a higher latitude and has the greatest capacity for accumulating triacylglycerols. A subtropical species, D. takahashii, which has no diapause in nature and is not cold-hardy, is unable to store the same quantities of triacylglycerols as temperate species.     

Diversity of seed storage protein patterns in wild peanut (Arachis,Fabaceae) species

55 accessions of wild peanuts (Arachis spp.) introduced from South America were analyzed for seed storage protein composition using SDS-PAGE electrophoresis. The objectives of the study were to evaluate variability within sect.Arachis and to classify taxa based on protein composition. 25 different band positions were resolved. Individual accessions had 11 to 18 bands which included the conarachin region (MW > 50 kD), two to five bands in the acidic arachin region (MW 38–49.9 kD), three to seven in the intermediate MW region (23 to 37.9 kD), two to five bands in the basic arachin region (18–22.9 kD), and one to three bands in the low MW protein region (14–17.9 kD). These data were utilized in a principal coordinate analysis based on the matrix of genetic distances between all pairs of the 55 accessions. Several groups of accessions conformed to expected species classification includingA. batizocoi, A. stenosperma, andA. monticola; whileA. duranensis, A. cardenasii, A. helodes, andA. correntina did not form good groups. The study showed that great diversity exists for protein profiles and seed storage proteins have potential for aiding species classification and for serving as markers for interspecific hybridization studies.     

A new species of Penicillium isolated from soil at the Kruger National Park in South Africa

A new species of Penicillium Link ex Fries is described and illustrated. It is represented by twenty six isolates recovered from soil at twenty six different locations at the southern part of the Paul Kruger National Park in South Africa. It clearly differs from all species of the genus described so far among the Biverticillata-Symmetrica Section, and it is therefore described and proposed as a new taxon: Penicillium krugerii sp. nov.     

The sequence of the isoepoxydon dehydrogenase gene of the patulin biosynthetic pathway in <Emphasis Type="Italic">Penicillium</Emphasis> species

Interest in species of the genus Penicillium is related to their ability to produce the mycotoxin patulin and to cause spoilage of fruit products worldwide. The sequence of the isoepoxydon dehydrogenase (idh) gene, a gene in the patulin biosynthetic pathway, was determined for 28 strains representing 12 different Penicillium species known to produce the mycotoxin patulin. Isolates of Penicillium carneum, Penicillium clavigerum, Penicillium concentricum, Penicillium coprobium, Penicillium dipodomyicola, Penicillium expansum, Penicillium gladioli, Penicillium glandicola, Penicillium griseofulvum, Penicillium paneum, Penicillium sclerotigenum and Penicillium vulpinum were compared. Primer pairs for DNA amplification and sequencing were designed from the P. griseofulvum idh gene (GenBank AF006680). The two introns present were removed from the nucleotide sequences, which were translated to produce the IDH sequences of the 12 species for comparison. Phylogenetic relationships among the species were determined from rDNA (ITS1, 5.8 S, ITS2 and partial sequence of 28S rDNA) and from the idh nucleotide sequences minus the two introns. Maximum parsimony analysis showed trees based on rDNA and idh sequences to be congruent. It is anticipated that the genetic information obtained in the present study will aid in the design of probes, specific for patulin biosynthetic pathway genes, to identify the presence of these mycotoxigenic fungi. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Differential response of fourTrichogramma species to low temperatures for short term storage

Differential response was noticed when 4 species ofTrichogramma egg parasitoids were stored at 2°, 5° and 10°C for 7 to 49 days. Pupal stage was found to be most appropriate for storage. Emergence ofTrichogramma achaeae Nagaraja and Nagarkatti andTrichogrammatoidea eldanae Viggiani was greater in comparison toTrichogramma chilonis Ishii andTrichogramma japonicum Ashmead at all 3 temperature regimes. Fecundity and longevity declined drastically after storage of 14 days at 2° and 5°C and 21 days at 10°C, when removed to room temperature. 10°C was found to be the best temperature for storage up to 49 days. Contribution No. 51004 of Biological Control Centre (NCIPM), Bangalore.     

Penicillium persicinum, a new griseofulvin, chrysogine and roquefortine C producing species from Qinghai province, China

A unique Penicillium isolate from Chinese soil with terverticillate penicilli and ellipsoidal to cylindrical smooth-walled conidia, produces, in addition to the common metabolite ergosterol, copious amounts of an unknown peach-red pigment and the following secondary metabolites: griseofulvin, dechlorogriseofulvin, lichexanthone, roquefortine C, roquefortine D, chrysogine, 2-pyrovoylaminobenzamide, 2-acetyl-quinazolin-4(3H)-one. This isolate, CBS 111235, is described as Penicillium persicinum sp. nov., which belongs to subgenus Penicillium section Chrysogena but is morphologically similar to P. italicum. On the basis of the production of secondary metabolites it resembles P. griseofulvum and P. coprophilum. Sequence data using part of the beta-tubulin gene showed that it is phylogenetically related to P. chrysogenum and P. aethiopicum in section Chrysogena with which it shares both secondary metabolites and ability to grow at 37 degrees C.     

Distribution of isoasparagine among different characean species

Thirteen species of Characeae were analyzed for their free amino acid contents. Large amounts of isoasparagine, accounting for 10 to 50% of the total free amino acids, were found in extracts fromChara (5 species including one unidentified),Nitellopsis (1 species), andLamprothamnium (1 species). In contrast, no isoasparagine was detected inNitella (5 species) andTolypella (1 species), except forN. flexilis in which as much as 40% of the free amino acids was isoasparagine. Other major amino acids found in the tested materials were Ala, Asp, Glu and Gln.     

Penicillium andAspergillus species in the habitats of allergy patients in the Tulsa, Oklahoma area

Penicillium andAspergillus have been recognized as important aeroallergens for more than 30 years, and are especially significant in indoor environments. There are over 400 species ofPenicillium andAspergillus combined, but there is little information on which species occur most frequently in the environment, or if each exhibits unique allergenic properties. A preliminary study showed no overlap between those species isolated from an outdoor site in Tulsa, Oklahoma and the species used in immunotherapy at allergy clinics in the Tulsa area. Pursuing this line of research, air samples were collected as three seasonal samples (over a 6 month period) in the homes or offices of ten allergy patients known to be allergic toPenicillium and/orAspergillus. Twenty three species ofPenicillium and 12 species ofAspergillus were identified from these samples through isolation, macroscopic, and microscopic examination.Penicillium corylophilum, P. glabrum, Aspergillus niger, andA. flavipes were the most abundant species isolated, supporting the data obtained in a preliminary study. At least in the Tulsa area, it appears that atopic patients are being tested and treated with extracts ofPenicillium andAspergillus species that are either not present or not abundant in the local indoor or outdoor environments. Additional research is necessary to determine if the environmental isolates share allergens with those species used in immunotherapy.     

Taxonomy ofPuccinia species causing rust diseases on sugarcane

A taxonomic revision ofPuccinia species causing rust diseases on sugarcane was conducted to clarify their morphological characteristics. Specimens including previously reported species,Puccinia melanocephala, P. kuehnii andPuccinia sp.sensu Muta, 1987, were collected in Japan and the Philippines and borrowed from various herbaria worldwide. Morphological characteristics of these specimens were examined under light and scanning electron microscopes. Comparative morphological studies of the specimens showed that rust fungi infecting sugarcane could be classified into two species,Puccinia melanocephala andP. kuehnii. Puccinia sp.sensu Muta was morphologically identical withP. kuehnii. Results of this study corroborate previous phylogenetic analysis results of D1/D2 regions of LSU rDNA gene. Contribution No. 157, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba     

Quantitative PCR analysis of selected Aspergillus, Penicillium and Paecilomyces species

A total of 65 quantitative PCR (QPCR) assays, incorporating fluorigenic 5' nuclease (TaqMan) chemistry and directed at the nuclear ribosomal RNA operon, internal transcribed spacer regions (ITS1 or ITS2) was developed and tested for the detection of selected Aspergillus, Penicillium and Paecilomyces species. The assays varied in specificity from species or subspecies to closely related species groups, subject to the amount of nucleotide sequence variation in the different organisms. A generic assay for all target species of Aspergillus, Penicillium and Paecilomyces was also developed and tested. Using a previously reported DNA extraction method, estimated conidia detection limits for target species ranged from less than one to several hundred per sample for the different assays. Conidia detection limits for non-target species were at least 1,000 fold higher in nearly all instances. The assays were used to analyze ten HVAC dust samples from different sources around the US. Total quantities of Aspergillus, Penicillium and Paecilomyces conidia in the samples, determined by the generic assay and the summed totals from the specific assays, were in general agreement, suggesting that all of the numerically dominant species in the samples were accounted for by the specific assays. QPCR analyses of these samples after spiking them with selected target organisms indicated that the enumeration results were within approximately a one-half log range of the expected values 95% of the time. Evidence is provided that the commonly used practices of enumerating Aspergillus and Penicillium as a single group or only by genus can be misleading in understanding the indoor populations of these organisms and their potential health risks.     

C-banding variation in the Moroccan oat species Avena agadiriana (2n=4x=28)

The C-banding technique was used to describe the chromosomes of a relatively recently-discovered Moroccan oat species, Avena agadiriana (2n=4x=28). A substantial amount of polymorphism for arm ratios and C-banding patterns was observed among five accessions of this species. However a common set of ten putatively homologous chromosomes was identifiable among the five accessions. The chromosomes of A. Agadiriana do not closely match those of any of the previously described diploid or tetraploid oat species in terms of their arm ratios and C-banding patterns. However, their overall C-banded appearance generally resembles the A/B/D groups of chromosomes of Avena species, rather than the more hetrochromatic C genomes. Implications of these findings in terms of chromosome evolution in the genus Avena are discussed.Contribution no. 95-490-J of the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS, USA     

Isolation and characterization of a cDNA clone encoding an auxin influx carrier in carnation cuttings. Expression in different organs and cultivars and its relationship with cold storage

Polar auxin transport (PAT) is necessary for the formation of adventitious roots in the base of leafy stem cuttings, as has been demonstrated in several studies in which the application of PAT inhibitors strongly inhibited the rooting of cuttings. However, unlike in the case of lateral roots, there is almost no information on the molecular mechanism that controls PAT in the formation of adventitious roots. A novel cDNA encoding an auxin influx carrier has been isolated and characterized from carnation (Dianthus caryophyllus) cuttings. The full length of DcAUX1 was obtained and the deduced aminoacid sequence revealed a high degree of identity with the corresponding auxin carrier proteins from several species. The expression of this gene depended on the organ, the carnation cultivar and the length of time cuttings had been stored in a cold chamber. As a rule, expression was higher in stem than in leaves, in the basal than in the first internode and in mature than in young leaves irrespective of the cultivar and the duration of the storage. This pattern of expression agrees with the results of a previous study showing that auxin from mature leaves was essential for rooting, while exogenous auxin applied to mature leaves was polarly transported in the stem and accumulated in the basal internode (the rooting zone). Variations in the expression observed during storage (depending of the cultivar) might be related to the variation in PAT and rooting reported in previous studies.     

不同林龄麻栎林地下部分生物量与碳储量研究

探讨不同林龄麻栎林地下部分根系的生物量与碳储量,为麻栎林的经营管理及碳汇管理等提供科学依据。以江苏省句容市不同林龄(幼龄林、中龄林、近熟林、成熟林)的麻栎林为研究对象,采用全根挖掘法获取麻栎各级根系及灌草层根系,并测定其生物量、碳含量,构建麻栎根系生物量模型,估算麻栎林地下部分根系碳储量及麻栎林群落碳储量。通过11种数学回归模型的比较,构建麻栎各级根系生物量幂回归模型,计算得到幼龄林、中龄林、近熟林、成熟林麻栎根系生物量分别为14.81t/hm~2、41.15t/hm~2、50.36t/hm~2、53.75t/hm~2,各级根系生物量大小顺序是:根桩粗根大根细根;灌木与草本植物根系生物量分别为0.48—1.71t/hm~2、0.13—0.60t/hm~2;不同林龄麻栎林群落根系生物量为15.42—56.06t/hm~2,且随林龄的增大而增大。麻栎根系碳含量大小顺序为:根桩粗根大根细根,且碳含量差异显著;灌木与草本植物根系碳含量分别为41.84%—43.79%、34.03%—38.48%,随林龄变化均无明显变化规律。麻栎林乔木根系碳储量随林龄增大而增大,幼龄林、中龄林、近熟林、成熟林根系碳储量分别为6.01t/hm~2、17.41t/hm~2、21.79t/hm~2、21.99t/hm~2;灌木与草本植物根系碳储量均随林龄增大而增大;幼龄林、中龄林、近熟林、成熟林群落根系碳储量分别为6.26t/hm~2、17.74t/hm~2、22.37t/hm~2、22.94t/hm~2,且乔木层灌木层草本层。麻栎林地下部分根系生物量与碳储量随林龄的增大而增大,幼龄林到近熟林生长过程中生物量与碳储量增加快速,近熟林后生物量与碳素积累缓慢,且与成熟林接近。     

Declining viability and lipid degradation during pollen storage

Declining viability of pollen during storage at 24° C in atmospheres of 40% relative humidity (RH) and 75% RH was studied, with special emphasis on lipid changes. Pollens of Papaver rhoeas and Narcissus poeticus, characterized by a high linolenic acid content, were compared with Typha latifolia pollen which has a low linolenic acid content. The rationale behind this was to answer the question of whether lipid peroxidation is involved in the rapid viability loss and reduced membrane integrity of, in particular, the unsaturated-lipid pollen types. Viability and membrane integrity degraded more rapidly at 75% RH than at 40% RH. All pollen species showed deesterification of acyl chains of lipids but no detectable peroxidation at both RH levels. Considerable amounts of lipid-soluble antioxidants were detected that did not degrade during storage. Free fatty acids and lysophospholipids were formed during storage, the effects of which on membranes are discussed. These degradation products were very prominent in the short-lived Papaver pollen. The loss of viability does coincide with phospholipid deesterification. A significant decrease of the phospholipid content occurred at 75% RH, but not at 40% RH. Based on compositional analyses of phospholipids and newly formed free fatty acids, it was concluded that the deesterification of acyl chains from the lipids occurred at random. We suggest that, due to the low water content of the pollen, free radicals rather than unspecific acyl hydrolases are involved in the deesterification process.     

Evolution of herkogamy and gynodioecy in Moroccan species ofRomulea (Iridaceae)

Herkogamy and gynodioecy were studied in the Moroccan species ofRomulea. Several types of herkogamy are shown to occur in the different species, with each type corresponding to a characteristic perianth size. The degree of differentiation between female and hermaphrodite morphs varies among the different gynodioecious species. Herkogamy is considered to have evolved prior to the development of the gynodioecious condition. An evolutionary interpretation is proposed based on the degree of herkogamy and of gynodioecy in the different species.     

Indigenous fish species for the control of Aedes aegypti in water storage tanks in Southern México

Because of inadequate supply of water, inhabitants of five villagesclose to Tapachula, Chiapas, México, store water in cement tanksthat support large populations of Aedes aegypti. Biologicalcontrol using indigenous fish species were studied to control A. aegypti larvae in thosecontainers, since other organisms used as biological control agents areexpensive and unfamiliar to inhabitants of those towns. Other measures(chemical or physical control) are expensive and time consuming. Fiveindigenous fish species, Lepisosteus tropicus (Gill)(Lepisosteiformes: Lepisosteidae), Astyanax fasciatus (Cuvier)(Cypriniformes: Characinidae), Brycon guatemalensis (Regan)(Cypriniformes: Characinidae), Ictalurus meridionalis(Günther) (Cypriniformes: Ictaluridae) and Poecilia sphenopsValenciennes (Cyprinodontiformes: Poeciliidae), currently used asmosquito control agents in the area were tested. Container indexes (ameasure of disease transmission potential) in the tested area werealways zero during the year of the study, independent of towns and fishspecies; this was significantly (P < 0.05) different from containerindexes prior to the test as well as from controls without fish. Nosignificant (P > 0.05) differences were recorded in the efficiency ofthe tested fish species feeding on A. aegypti larvae. Our resultsshow that all tested fish species can be considered as good biologicalagents for controlling A. aegypti larvae in Southern Mexico.     

Expression of anthocyanin biosynthesis pathway genes in red and white grapes

The expression of seven genes from the anthocyanin biosynthesis pathway was determined in different tissues of Shiraz grapevines. All of the tissues contained proanthocyanidins, but only the berry skin accumulated anthocyanins. In most tissues, all of the flavonoid genes except UDP glucose-flavonoid 3-o-glucosyl transferase (UFGT) were expressed, but UFGT expression was only detected in berry skin. Similar patterns of expression were observed in the skin of other red grapes. In white grapes, UFGT expression was not detected. White grape cultivars appear to lack anthocyanins because they lack UFGT, although they also had decreased expression of other flavonoid pathway genes.