CS and CCP are effective substrates for the production of fruit bodies and bioactive compounds by C. militaris. This study provides a new approach to decreasing the cost of C. militaris cultivation and dealing with these agricultural wastes. © 2016 Society of Chemical Industry.

Costus speciosus (Koen.) Smith has been an important medicinal agent in the various traditional and folk systems of medicine in southern China. In September 2018, leaf spot disease was detected on C. speciosus plants in Chengmai County, Hainan Province. A survey of C. speciosus plants revealed that the disease caused serious damage during the typhoon season of September to November in Hainan Province, with 80 to85% incidence in plants. Early symptoms were yellow-to-brown, irregular-shaped lesions on the leaf margin or tip. After several days, lesions expanded along the mid-vein until the entire leaf was destroyed. Then, the infected leaves turned gray brown, leading to defoliation. Heavily infected leaves became dry and died. The pathogen was consistently isolated from the lesions and pure single-spore cultures were obtained. Twenty pieces of diseased leaf samples were plated and seven pieces yielded fungal colonies after 5 to 6 days of incubation at 25 °C. Colonies on potato dextrose agar (PDA) were white and later became gray to black. Conidia were unicellular, terminal, black, elliptical that measured 10 to 13 (length) × 12 to 16 (width) μm (n=30), growing aerial mycelium covering the entire petri dish (9 cm in diameter). The morphological characteristics and measurements of this fungal pathogen matched the previous descriptions of Nigrospora oryzae (Wang et al. 2017). To confirm identity the internal transcribed spacer (ITS) region of the ribosomal DNA was amplified using primers ITS1/ITS4 (Mills, P. R., et al. 1992), and the 530-bp product (GenBank Accession No. MK280694) of the ITS showed 99% sequence identity to N. oryzae isolates TLFa21 (GenBank Accession No. MH790146) and xsd08022 (GenBank Accession No. EU918714). Pathogenicity tests were conducted. Three leaves of three C. speciosus plants were wounded and inoculated with mycelial plugs (5×5mm) , and an additional five plants were inoculated with PDA plugs as a control. All plants were placed in the field and temperature ranged from 23 to 29°C. Ten days after inoculation, all the inoculated plants showed typical leaf spot symptoms, a yellow-to-brown mildew appeared at the points of inoculation. No symptoms were observed on the controls. The fungus was re-isolated from the infected tissues, fulfilling Koch’s postulates. N. oryzae was previously reported as the causal agent of leaf spot on cotton (Zhang. et al. 2012), dendrobium candidum (Wu. et al. 2014) and Aloe vera (Zhai. et al. 2013) in china. To our knowledge, this is the first report of leaf spot of C. speciosus caused by N. oryzae in China. The project was partially founded by Hainan Provincial Research Institute of technology development projects (Screening and application of endophytic bacteria with high resistance to Fusarium Wilt of Sauropus androgynus), Hainan Provincial Key Laboratory for Vegetables and Biology，Hainan Provincial Engineering Research Center for Melon and Vegetable Breeding, Major scientific and technological projects in Hainan Province(ZDKJ2017001)，Third Survey and Collection of Crop Germplasm Resources in China, Collection, identification and preservation of pathogenic bacteria of inverted season vegetable in Hainan. References: L. F. Zhai., et al.2013. Plant Dis.97:1256 L. X. Zhang., et al.2012. Plant Dis.102:2029 J. B. Wu., et al.2014. Plant Dis.98:996 Mills, P. R., et al. 1992. FEMS Microbiol Lett. 98:137-144 Wang et al. 2017. Persoonia 39: 118-142

For matching the new fungal nomenclature to abolish pleomorphic names for a fungus, a genus Pseudocercospora s. str. was suggested to host holomorphic Pseudocercosproa fungi. But the Pseudocercosproa fungi need extra phylogenetic loci to clarify their taxonomy and diversity for their existing and coming species. Internal transcribed spacer 2 (ITS2) secondary structures have been promising in charactering species phylogeny in plants, animals and fungi. In present study, a conserved model of ITS2 secondary structures was confirmed on fungi in Pseudocercospora s. str. genus using RNAshape program. The model has a typical eukaryotic four-helix ITS2 secondary structure. But a single U base occurred in conserved motif of U-U mismatch in Helix 2, and a UG emerged in UGGU motif in Helix 3 to Pseudocercospora fungi. The phylogeny analyses based on the ITS2 sequence-secondary structures with compensatory base change characterizations are able to delimit more species for Pseudocercospora s. str. than phylogenic inferences of traditional multi-loci alignments do. The model was employed to explore the diversity of endophytic Pseudocercospora fungi in poplar trees. The analysis results also showed that endophytic Pseudocercospora fungi were diverse in species and evolved a specific lineage in poplar trees. This work suggested that ITS2 sequence-structures could become as additionally significant loci for species phylogenetic and taxonomic studies on Pseudocerospora fungi, and that Pseudocercospora endophytes could be important roles to Pseudocercospora fungi's evolution and function in ecology.

Kalanchoe pinnata (Lam.) Pers. [syn.: Bryophyllum pinnatum (Lam.) Oken] is an important medicinal agent in southern China. The succulent leaves of this plant are used in the treatment of cholera, bruises, uri­nary diseases and whitlow. In Oct. 2019, leaf spots were detected on K. pinnata plants in Chengmai County, Hainan Province, China. Lesions with brown to black margins were irregularly shaped and associated with leaf margins. Spots coalesced to form larger lesions (Fig. S1-A), with black pycnidia present in more mature lesions. Symptomatic K. pinnata were found with 10-20% incidence during the humid winters of Hainan Province. Leaf tissues of 10 symptomatic plants were collected and surface sterilized in 70% ETOH for 30s, 0.1% HgCl2 for 30 s, rinsed 3x with sterile distilled water for 30s, placed on potato dextrose agar (PDA) amended with 30mg/L of kanamycin sulfate, and incubated at 25°C in the dark for 3-5 days. Four fungal isolates were obtained using a single-spore isolation method. The colonies were floccose, dense, and white with forming on older colonies grown on PDA (Fig. S1-B-1&2). Alpha conidia exuded from ostiole, rostrate, long-beaked pycnidia in creamy-to-yellowish drops. Alpha conidia were hyaline, ellipsoidal, separated and averaged 6.3μm (SD ± 1.13) long × 1.9μm (SD ± 0.33) wide (n=50). Beta conidia were not seen. The morphological characteristics matched the previous description of Diaporthe longicolla (syn. Phomopsis longicolla) (Hobbs et al. 1985). Mycelial genomic DNA of the representative isolate LDSG3-2 was extracted as template. The internal transcribed spacer (ITS) , translation elongation factor 1α gene (TEF) and β-tubulin (TUB2) regions were amplified. These loci were amplified using primer pairs ITS4/ITS5 (White, et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999) and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. A BLAST search of GenBank showed ITS (MN960195), TEF (MN974483) and TUB2 (MN974482) sequences of the isolate were 99%, 100%, and 99% homologous with D. longicolla strains DL11 (MF125048, 557/563 bp), D55 (MN584792, 347/347 bp) and DPC-HOH-32 (MK161506, 502/504 bp). Maximum likelihood trees based on concatenated nucleotide sequences of the three genes were constructed using MEGA 7.0, and bootstrap values indicated the isolate was D. longicolla (Fig. S1-D). Pathogenicity testing was performed using isolate LDSG3-2 by depositing 5µl droplets of a conidial suspension (1 × 106 ml-1) into 5 artificially wounded leaves (using a sterile needle) of 10 healthy 3-month-old K. pinnata plants. An equal number of artificially wounded control leaves were inoculated with sterile water to serve as a negative control. The test was conducted three times. Plants were kept at 25°C in 80% relative humidity and observed for symptoms. Two weeks after inoculation, no symptoms were observed on control plants (Fig. S1-C-1) and all inoculated plants showed symptoms (Fig. S1-C-2) similar to those observed in the field. The fungus was re-isolated from the infected tissues and showed the same cultural and morphological characteristics of the strain inoculated and could not be isolated from the controls fulfilling Koch’s postulates. To our knowledge, this is the first report of leaf spot on K. pinnata caused by D. longicolla in China. This disease is of concern since Phomopsis diseases are common in K. pinnata fields and can cause significant reduction in yield. References: White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. DOI: 10.1016/0167-7799(90)90215-J Carbone, I., and Kohn, L. M. 1999. Mycologia. 91:553. DOI: 10.2307/3761358 Glass, N. L., and Donaldson, G. C. 1995. Appl. Environ. Microbiol. 61:1323. DOI: 10.1002/bit.260460112 Hobbs, T. W. et al. 1985. Mycologia. 77: 535. DOI: 10.2307/3793352