The brlA Gene Deletion Reveals That Patulin Biosynthesis Is Not Related to Conidiation in Penicillium expansum.

Dissemination and survival of ascomycetes is through asexual spores. The brlA gene encodes a C2H2-type zinc-finger transcription factor, which is essential for asexual development. Penicillium expansum causes blue mold disease and is the main source of patulin, a mycotoxin that contaminates apple-based food. A P. expansum PeΔbrlA deficient strain was generated by homologous recombination. In vivo, suppression of brlA completely blocked the development of conidiophores that takes place after the formation of coremia/synnemata, a required step for the perforation of the apple epicarp. Metabolome analysis displayed that patulin production was enhanced by brlA suppression, explaining a higher in vivo aggressiveness compared to the wild type (WT) strain. No patulin was detected in the synnemata, suggesting that patulin biosynthesis stopped when the fungus exited the apple. In vitro transcriptome analysis of PeΔbrlA unveiled an up-regulated biosynthetic gene cluster (PEXP_073960-PEXP_074060) that shares high similarity with the chaetoglobosin gene cluster of Chaetomium globosum. Metabolome analysis of PeΔbrlA confirmed these observations by unveiling a greater diversity of chaetoglobosin derivatives. We observed that chaetoglobosins A and C were found only in the synnemata, located outside of the apple, whereas other chaetoglobosins were detected in apple flesh, suggesting a spatial-temporal organization of the chaetoglobosin biosynthesis pathway.

Fusarium xyrophilum, sp. nov., a member of the Fusarium fujikuroi species complex recovered from pseudoflowers on yellow-eyed grass (Xyris spp.) from Guyana.

We report on the discovery and characterization of a novel Fusarium species that produced yellow-orange pseudoflowers on Xyris spp. (yellow-eyed grass; Xyridaceae) growing in the savannas of the Pakaraima Mountains of western Guyana. The petaloid fungal structures produced on infected plants mimic host flowers in gross morphology. Molecular phylogenetic analyses of full-length RPB1 (RNA polymerase largest subunit), RPB2 (RNA polymerase second largest subunit), and TEF1 (elongation factor 1-α) DNA sequences mined from genome sequences resolved the fungus, described herein as F. xyrophilum, sp. nov., as sister to F. pseudocircinatum within the African clade of the F. fujikuroi species complex. Results of a polymerase chain reaction (PCR) assay for mating type idiomorph revealed that single-conidial isolates of F. xyrophilum had only one of the MAT idiomorphs (MAT1-1 or MAT1-2), which suggests that the fungus may have a heterothallic sexual reproductive mode. BLASTn searches of whole-genome sequence of three strains of F. xyrophilum indicated that it has the genetic potential to produce secondary metabolites, including phytohormones, pigments, and mycotoxins. However, a polyketide-derived pigment, 8-O-methylbostrycoidin, was the only metabolite detected in cracked maize kernel cultures. When grown on carnation leaf agar, F. xyrophilum is phenotypically distinct from other described Fusarium species in that it produces aseptate microconidia on erect indeterminate synnemata that are up to 2 mm tall and it does not produce multiseptate macroconidia.

Morphology and multigene phylogeny of Talaromycesamyrossmaniae, a new synnematous species belonging to the section Trachyspermi from India.

A new Talaromyces species, T.amyrossmaniae, isolated from decaying fruit and litter of Terminalia bellerica, is described and illustrated. On the natural substrate, the new species produces determinate synnemata, with a well-defined, vivid orange red to orange red cylindrical stipe, and a greyish green capitulum. Conidiophores are typically biverticillate, or sometimes have subterminal branches, with acerose phialides that produce globose to subglobose, smooth to slightly roughened conidia. Multigene phylogenetic analyses based on the internal transcribed spacer region (ITS), and partial sequences of ß-tubulin (BenA), calmodulin (CaM), and DNA directed RNA polymerase second large subunit (RPB2) genes, along with morphological characterization, revealed that these isolates are distinct and form a unique lineage of Talaromyces in section Trachyspermi, closely allied to T.aerius, T.albobiverticillius, T.heiheensis, T.erythromellis, and T.solicola. The new species T.amyrossmaniae is the first species in section Trachyspermi with determinate synnemata.

Braunomyces dictyosporus gen. sp. nov. from Vietnam.

The generic name Braunomyces (ascomycetes, asexual morph), with B. dictyosporus as type species, is described, illustrated and discussed, based on material collected in Vietnam on leaf debris of an unidentified broadleaved tree. The new genus is well characterised and quite distinct from other synnematous and non-synnematous dematiaceous hyphomycete genera by its unique combination of traits, viz. determinate synnematous conidiomata, integrated, terminal and intercalary conidiogenous cells with one to several conidiogenous vesicles becoming cupulate with age, tretic conidiogenesis, and mostly cruciately septate, solitary dictyoconidia.

Synnemata Production Using Silkworm Variety, Female Yangwonjam by Isaria tenuipes.

This study was conducted to confirm the possible use of female Yangwonjam as a host for synnemata production of Isaria tenuipes in eight local areas in Korea. Silkworm pupation rate, infection rate and synnemata characteristics of I. tenuipes were examined. Normal silkworms had a higher pupation rate than silkworms inoculated with I. tenuipes. The pupae survival percentage of normal silkworm in cocoons was 92.5~97.6%, whereas it ranged from 91.1~95.6% in silkworms sprayed with I. tenuipes. Female Yangwonjam showed the highest survival percentage at 97.6% among the silkworm varieties tested. I. tenuipes infection rate of larvae of 5th instar newly-exuviated silkworms was 89.2~90.7% in the spring rearing season and 98.2~99.3% in the autumn rearing season. Synnemata production of I. tenuipes was excellent in female Yangwonjam with an incidence rate of 98.0% followed by male Yangwonjam (94.1%) and Baegokjam (93.3%) in the spring rearing season. Synnemata living weight ranged from 1.44~0.94 g in the spring rearing season. The female Yangwonjam had the heaviest synnemata weight (1.44 g) in the spring rearing season. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and were similar in shape and color to wild synnemata collected in Korea.

Synnemata Production Using Silkworm Variety, Female Yangwonjam by Isaria tenuipes

This study was conducted to confirm the possible use of female Yangwonjam as a host for synnemata production of Isaria tenuipes in eight local areas in Korea. Silkworm pupation rate, infection rate and synnemata characteristics of I. tenuipes were examined. Normal silkworms had a higher pupation rate than silkworms inoculated with I. tenuipes. The pupae survival percentage of normal silkworm in cocoons was 92.5~97.6%, whereas it ranged from 91.1~95.6% in silkworms sprayed with I. tenuipes. Female Yangwonjam showed the highest survival percentage at 97.6% among the silkworm varieties tested. I. tenuipes infection rate of larvae of 5th instar newly-exuviated silkworms was 89.2~90.7% in the spring rearing season and 98.2~99.3% in the autumn rearing season. Synnemata production of I. tenuipes was excellent in female Yangwonjam with an incidence rate of 98.0% followed by male Yangwonjam (94.1%) and Baegokjam (93.3%) in the spring rearing season. Synnemata living weight ranged from 1.44~0.94 g in the spring rearing season. The female Yangwonjam had the heaviest synnemata weight (1.44 g) in the spring rearing season. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and were similar in shape and color to wild synnemata collected in Korea.

Cultural Characteristics and Fruiting Body Production in Cordyceps bassiana.

Single ascospore isolates of Cordyceps bassiana were observed for their colony pigmentation on Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates and were inoculated in a brown rice medium for production of fruiting bodies. Colony pigmentation did not show any relationship with perithecial stromata formation. The isolates were also grown on opposite sides of SDAY agar plates and were observed for vegetative compatibility. Neither vegetative compatibility nor perithecial stromata could be found to be related to each other. It was concluded that fertile fruiting body production was independent of colony pigmentation and vegetative compatibility. Synnemata formation was found to be more common than perithecial stromata formation. This might be due to its highly conidiogenous anamorphic stage, i.e., Beauveria bassiana.

Breeding of a Silkworm Variety for Synnemata Production of Isaria tenuipes.

This study was conducted out to select a silkworm variety suitable for synnemata production of Isaria tenuipes. Four kinds of the mulberry silkworm varieties, Bombyx mori, were hybridized using a Japanese parental line and a Chinese parental line, and used to test for synemata formation in I. tenuipes. The larval period of normal silkworms was 22 hr longer than the silkworms inoculated with this fungus. Among the silkworm varieties tested, Hachojam had the shortest larval period with 23.02 days. The non-cocooning silkworm had a shorter larval period than the cocoon producing silkworms. The pupation rate of normal silkworms was about 9% higher than that of silkworms sprayed with I. tenuipes. Hachojam had the highest infection rate at 99.8%, but no significant difference was observed for the infection rate by silkworm variety. The production of synnemata was the best in JS171 × CS188 with an incidence rate of 99.3%, followed by Hachojam, and Chugangjam. The synnemata produced from Hachojam were the heaviest and showed white or milky-white in color.

Cultural Characteristics and Fruiting Body Production in Cordyceps bassiana

Single ascospore isolates of Cordyceps bassiana were observed for their colony pigmentation on Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates and were inoculated in a brown rice medium for production of fruiting bodies. Colony pigmentation did not show any relationship with perithecial stromata formation. The isolates were also grown on opposite sides of SDAY agar plates and were observed for vegetative compatibility. Neither vegetative compatibility nor perithecial stromata could be found to be related to each other. It was concluded that fertile fruiting body production was independent of colony pigmentation and vegetative compatibility. Synnemata formation was found to be more common than perithecial stromata formation. This might be due to its highly conidiogenous anamorphic stage, i.e., Beauveria bassiana.

Breeding of a Silkworm Variety for Synnemata Production of Isaria tenuipes

This study was conducted out to select a silkworm variety suitable for synnemata production of Isaria tenuipes. Four kinds of the mulberry silkworm varieties, Bombyx mori, were hybridized using a Japanese parental line and a Chinese parental line, and used to test for synemata formation in I. tenuipes. The larval period of normal silkworms was 22 hr longer than the silkworms inoculated with this fungus. Among the silkworm varieties tested, Hachojam had the shortest larval period with 23.02 days. The non-cocooning silkworm had a shorter larval period than the cocoon producing silkworms. The pupation rate of normal silkworms was about 9% higher than that of silkworms sprayed with I. tenuipes. Hachojam had the highest infection rate at 99.8%, but no significant difference was observed for the infection rate by silkworm variety. The production of synnemata was the best in JS171 x CS188 with an incidence rate of 99.3%, followed by Hachojam, and Chugangjam. The synnemata produced from Hachojam were the heaviest and showed white or milky-white in color.

Complete genome sequence of Actinosynnema mirum type strain (101).

Actinosynnema mirum Hasegawa et al. 1978 is the type species of the genus, and is of phylogenetic interest because of its central phylogenetic location in the Actino-synnemataceae, a rapidly growing family within the actinobacterial suborder Pseudo-nocardineae. A. mirum is characterized by its motile spores borne on synnemata and as a producer of nocardicin antibiotics. It is capable of growing aerobically and under a moderate CO(2) atmosphere. The strain is a Gram-positive, aerial and substrate mycelium producing bacterium, originally isolated from a grass blade collected from the Raritan River, New Jersey. Here we describe the features of this organism, together with the complete genome sequence and annotation. This is the first complete genome sequence of a member of the family Actinosynnemataceae, and only the second sequence from the actinobacterial suborder Pseudonocardineae. The 8,248,144 bp long single replicon genome with its 7100 protein-coding and 77 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project.

Stable Formation of Fruiting Body in Cordyceps bassiana.

In order to breed a Cordyceps bassiana isolate that stably forms fruiting body in artificial cultivation, isolates derived from subculturing and single spores were tested through mating. From C. bassiana EFCC 783, three subcultured isolates EFCC 2830, EFCC 2831 and EFCC 2832 were obtained and fourteen single conidial isolates were obtained from these three subcultured isolates. Two different morphological types were found in the fourteen single conidial isolates. One type was able to form synnemata and another type was not able to form synnemata. Since switch of morphological type was not observed despite their continuous subculturing, cross was performed between the two types and the formation of fruiting body was examined. Ascospores were obtained from a selected fruiting body formed by hybrid of the cross. Self-cross and combinational cross of the ascospore-derived isolates generated hybrids that stably produce high quality fruiting body in artificial media.

Stable Formation of Fruiting Body in Cordyceps bassiana

In order to breed a Cordyceps bassiana isolate that stably forms fruiting body in artificial cultivation, isolates derived from subculturing and single spores were tested through mating. From C. bassiana EFCC 783, three subcultured isolates EFCC 2830, EFCC 2831 and EFCC 2832 were obtained and fourteen single conidial isolates were obtained from these three subcultured isolates. Two different morphological types were found in the fourteen single conidial isolates. One type was able to form synnemata and another type was not able to form synnemata. Since switch of morphological type was not observed despite their continuous subculturing, cross was performed between the two types and the formation of fruiting body was examined. Ascospores were obtained from a selected fruiting body formed by hybrid of the cross. Self-cross and combinational cross of the ascospore-derived isolates generated hybrids that stably produce high quality fruiting body in artificial media.