Comprehensive Transcriptomic Analysis of Cordyceps militaris Cultivated on Germinated Soybeans.

The ascomycete fungus Cordyceps militaris infects lepidopteran larvae and pupae and forms characteristic fruiting bodies. Owing to its immune-enhancing effects, the fungus has been used as a medicine. For industrial application, this fungus can be grown on geminated soybeans as an alternative protein source. In our study, we performed a comprehensive transcriptomic analysis to identify core gene sets during C. militaris cultivation on germinated soybeans. RNA-Seq technology was applied to the fungal cultures at seven-time points (2, 4, and 7-day and 2, 3, 5, 7-week old cultures) to investigate the global transcriptomic change. We conducted a time-series analysis using a two-step regression strategy and chose 1460 significant genes and assigned them into five clusters. Characterization of each cluster based on Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed that transcription profiles changed after two weeks of incubation. Gene mapping of cordycepin biosynthesis and isoflavone modification pathways also confirmed that gene expression in the early stage of GSC cultivation is important for these metabolic pathways. Our transcriptomic analysis and selected genes provided a comprehensive molecular basis for the cultivation of C. militaris on germinated soybeans.

Genome-wide in silico identification of phospholipase D (PLD) gene family from Corchorus capsularis and Corchorus olitorius: reveals their responses to plant stress.

BACKGROUND: Plant grows in nature facing various types of abiotic stresses for their normal growth and development. During abiotic stress, plants evolve different types of mechanisms to survive in a hostile environment. Phospholipase D (PLD) plays important role in the regulation of diverse cellular processes including stress responses in plants. Member of PLD genes are well studied in different model plants; however, their functions in the jute are not clear yet. RESULT: In the present study, a total of 12 and 11 PLD genes were identified in the genome of C. capsularis and C. olitorius, respectively. The presence of the two conserved HKD motifs in PLD genes except for CoPLDδ-2 in jute suggests their strong lipase activity. Twenty different motifs were found in the identified PLD genes, and PLD-ß1, PLD-γ1, and all members of PLD-δ1 of both jute species contained the highest number of motifs. Phylogenetic analysis showed the close evolutionary relationship among the five groups of jute PLD proteins along with the PLD proteins from Arabidopsis. Tissue-specific expression pattern of PLDα1-2, PLD-α2, PLDß1, PLDγ1, and PLDδ1 of two jute species suggested their involvement in plant growth and development. However, the expression pattern of PLDα1-2, PLDα1-3, PLD-α4, PLDδ1, and PLDδ3 indicated their association during waterlogging stress. In addition, PLD-α2, PLDß1, and PLDδ2 seemed to be involved in drought stress as well as salinity stress. CONCLUSION: This genome-wide identification of jute PLD genes from C. capsularis and C. olitorius will help to further functional characterization of the PLD genes for developing stress-tolerant jute variety.

The Membrane-Bound Protein, MoAfo1, Is Involved in Sensing Diverse Signals from Different Surfaces in the Rice Blast Fungus.

To establish an infection, fungal pathogens must recognize diverse signals from host surfaces. The rice blast fungus, Magnaporthe oryzae, is one of the best models studying host-pathogen interactions. This fungus recognizes physical or chemical signals from the host surfaces and initiates the development of an infection structure called appressorium. Here, we found that protein MoAfo1(appressorium formation, MGG_10422) was involved in sensing signal molecules such as cutin monomers and long chain primary alcohols required for appressorium formation. The knockout mutant (ΔMoafo1) formed a few abnormal appressoria on the onion and rice sheath surfaces. However, it produced normal appressoria on the surface of rice leaves. MoAfo1 localized to the membranes of the cytoplasm and vacuole-like organelles in conidia and appressoria. Additionally, the ΔMoafo1 mutant showed defects in appressorium morphology, appressorium penetration, invasive growth, and pathogenicity. These multiple defects might be partially due to failure to respond properly to oxidative stress. These findings broaden our understanding of the fungal mechanisms at play in the recognition of the host surface during rice blast infection.

Genome-wide identification of F-box proteins in Macrophomina phaseolina and comparison with other fungus.

BACKGROUND: In fungi, like other eukaryotes, protein turnover is an important cellular process for the controlling of various cellular functions. The ubiquitin-proteasome pathway degrades some selected intracellular proteins and F-box proteins are one of the important components controlling protein degradation. F-box proteins are well studied in different model plants however, their functions in the fungi are not clear yet. This study aimed to identify the genes involved in protein degradation for disease development in the Macrophomina phaseolina fungus. RESULTS: In this research, in silico studies were done to understand the distribution of F-box proteins in pathogenic fungi including Macrophomina phaseolina fungus. Genome-wide analysis indicates that M. phaseolina fungus contained thirty-one F-box proteins throughout its chromosomes. In addition, there are 17, 37, 16, and 21 F-box proteins have been identified from Puccinia graminis, Colletotrichum graminicola, Ustilago maydis, and Phytophthora infestans, respectively. Analyses revealed that selective fungal genomes contain several additional functional domains along with F-box domain. Sequence alignment showed the substitution of amino acid in several F-box proteins; however, gene duplication was not found among these proteins. Phylogenetic analysis revealed that F-box proteins having similar functional domain was highly diverse form each other showing the possibility of various function. Analysis also found that MPH_00568 and MPH_05531 were closely related to rice blast fungus F-box protein MGG_00768 and MGG_13065, respectively, may play an important role for blast disease development. CONCLUSION: This genome-wide analysis of F-box proteins will be useful for characterization of candidate F-box proteins to understand the molecular mechanisms leading to disease development of M. phaseolina in the host plants.

Wheat Blast: A New Fungal Inhabitant to Bangladesh Threatening World Wheat Production.

World wheat production is now under threat due to the wheat blast outbreak in Bangladesh in early March 2016. This is a new disease in this area, indicating the higher possibility of this pathogen spreading throughout the Asia, the world's largest wheat producing area. Occurrence of this disease caused ~3.5% reduction of the total wheat fields in Bangladesh. Its economic effect on the Bangladesh wheat market was little because wheat contributes to 3% of total cereal consumption, among which ~70% have been imported from other countries. However, as a long-term perspective, much greater losses will occur once this disease spreads to other major wheat producing areas of Bangladesh, India, and Pakistan due to the existing favorable condition for the blast pathogen. The wheat blast pathogen belongs to the Magnaporthe oryzae species complex causing blast disease on multiple hosts in the Poaceae family. Phylogenetic analysis revealed that the Bangladesh outbreak strains and the Brazil outbreak strains were the same phylogenetic lineage, suggesting that they might be migrated from Brazil to Bangladesh during the seed import. To protect wheat production of Bangladesh and its neighbors, several measures including rigorous testing of seed health, use of chemicals, crop rotation, reinforcement of quarantine procedures, and increased field monitoring should be implemented. Development of blast resistant wheat varieties should be a long-term solution and combination of different methods with partial resistant lines may suppress this disease for some time.

Regulation of cellular diacylglycerol through lipid phosphate phosphatases is required for pathogenesis of the rice blast fungus, Magnaporthe oryzae.

Considering implication of diacylglycerol in both metabolism and signaling pathways, maintaining proper levels of diacylglycerol (DAG) is critical to cellular homeostasis and development. Except the PIP2-PLC mediated pathway, metabolic pathways leading to generation of DAG converge on dephosphorylation of phosphatidic acid catalyzed by lipid phosphate phosphatases. Here we report the role of such enzymes in a model plant pathogenic fungus, Magnaporthe oryzae. We identified five genes encoding putative lipid phosphate phosphatases (MoLPP1 to MoLPP5). Targeted disruption of four genes (except MoLPP4) showed that MoLPP3 and MoLPP5 are required for normal progression of infection-specific development and proliferation within host plants, whereas MoLPP1 and MoLPP2 are indispensable for fungal pathogenicity. Reintroduction of MoLPP3 and MoLPP5 into individual deletion mutants restored all the defects. Furthermore, exogenous addition of saturated DAG not only restored defect in appressorium formation but also complemented reduced virulence in both mutants. Taken together, our data indicate differential roles of lipid phosphate phosphatase genes and requirement of proper regulation of cellular DAGs for fungal development and pathogenesis.