The secret password: Cell death-inducing proteins in filamentous phytopathogens - As versatile tools to develop disease-resistant crops.

Cell death-inducing proteins (CDIPs) are some of the secreted effector proteins manifested by filamentous oomycetes and fungal pathogens to invade the plant tissue and facilitate infection. Along with their involvement in different developmental processes and virulence, CDIPs play a crucial role in plant-pathogen interactions. As the name implies, CDIPs cause necrosis and trigger localised cell death in the infected host tissues by the accumulation of higher concentrations of hydrogen peroxide (H2O2), oxidative burst, accumulation of nitric oxide (NO), and electrolyte leakage. They also stimulate the biosynthesis of defense-related phytohormones such as salicylic acid (SA), jasmonic acid (JA), abscisic acid (ABA), and ethylene (ET), as well as the expression of pathogenesis-related (PR) genes that are important in disease resistance. Altogether, the interactions result in the hypersensitive response (HR) in the host plant, which might confer systemic acquired resistance (SAR) in some cases against a vast array of related and unrelated pathogens. The CDIPs, due to their capability of inducing host resistance, are thus unique among the array of proteins secreted by filamentous plant pathogens. More interestingly, a few transgenic plant lines have also been developed expressing the CDIPs with added resistance. Thus, CDIPs have opened an interesting hot area of research. The present study critically reviews the current knowledge of major types of CDIPs identified across filamentous phytopathogens and their modes of action in the last couple of years. This review also highlights the recent breakthrough technologies in studying plant-pathogen interactions as well as crop improvement by enhancing disease resistance through CDIPs.

High black carbon episodes over a polluted metropolis near the land-sea boundary and their impact on associated atmospheric dynamics.

The present investigation outlines the crucial factors that influence the black carbon (BC) concentrations over a polluted metropolis, Kolkata (22.57° N, 88.37° E), India. Located in the eastern part of the Indo Gangetic Plain (IGP) outflow region and close to the land-ocean boundary, Kolkata is subject to contrasting seasonal maritime airflow from the Bay of Bengal and continental air mass from the IGP and Tibetan plateau region, which modulates the local concentration of BC. The origin of aerosol transport and associated atmospheric dynamics with high and low BC activities over Kolkata are examined during 2012-2015 using data from multi-technique sources which include measurements of ground-based instruments of aethalometer and multi-frequency microwave radiometer, reanalysis data from ERA-5 and MEERA-2, and model outputs from HYPSLIT back trajectory model simulations. The study highlights the control of IGP wind inflow on the occurrence of anomalous enhancements in BC concentration during weekends and holidays when local emissions are low. High BC events are associated with enhanced atmospheric heating below the boundary layer (2000 m) and significant negative surface radiative forcing. The response of the boundary layer to high and low BC episodes, shown in the diurnal variation in comparison with the seasonal mean, is investigated. Dominant suppression of morning and night-time boundary layer height is observed on high BC days. During the daytime in pre-monsoon, post-monsoon, and winter seasons, boundary layer height peaks are found to be strongly controlled by high BC episode occurrences as obtained from the hourly data of ERA-5.

Metabolite profiling reveals overexpression of the global regulator, MoLAEA leads to increased synthesis of metabolites in Magnaporthe oryzae.

AIMS: To study the altered metabolic pathways and metabolites produced in overexpression and knockdown mutants of a global regulator named MoLAEA, which was recently found to regulate the expression of the genes involved in secondary metabolism in one of the most destructive plant pathogens, Magnaporthe oryzae. METHODS AND RESULTS: Mass spectrometry-based global untargeted metabolomic profiling was used to identify altered metabolites. Metabolites were extracted from the mutant strains of MoLAEA using two extraction methods viz., aqueous and organic extraction and data acquired using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive and negative polarities. Levels of metabolites involved in various biological pathways such as amino acid as well as polyamine biosynthesis, fatty acid and pyrimidine metabolism showed a remarkable change in the mutant strains. Interestingly, metabolites involved in stress responses were produced in higher quantities in the overexpression strain, whereas certain overproduced metabolites were associated with distinctive phenotypic changes in the overexpression strain compared with the wild type. Further, the expression of several genes involved in the stress responses was found to have higher expression in the overexpression strain. CONCLUSIONS: The global regulator MoLAEA is involved in secondary metabolism in the plant pathogen M. oryzae such that the mutant strains showed an altered level of several metabolites involved in the biosynthesis pathways compared with the wild type. Also, metabolites involved in stress responses were overproduced in the overexpression strain and this can be seen in the higher growth in media amended with stress-inducing agents or a higher expression of genes involved in stress response in the overexpression strain compared with the wild type. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of metabolite profiling relative to the global regulation of secondary metabolism in M. oryzae, where secondary metabolism is poorly understood. It opens up avenues for more relevant investigations on the genetic regulation of several of the metabolites found in the analysis, which have not been previously characterized in M. oryzae.

Fluoride release by restorative materials after applying surface coating agents.

The prompt use of an enamel surface covering reagent is advised to safeguard the dental restorative substance from mishaps. Therefore, it is of interest to assess the fluoride emitting capabilities of standard GIC, and Zirconomer cement together with surface coverings and without surface coverings. The conventional GIC cement was part of experimental category A while Zirconomer cement was part of category B. For every experimental categories, a set of sixty brass mould prototypes in the form of disc with dimensions: diameter (6±0.1mm) and thickness (2±0.1 mm) were created and subsequently covered with Teflon strip in accordance with the package recommendations. Also, for both experimental categories, such pellets were randomly allocated to three sub-categories of 20 each. For one category petroleum jelly was administered with a cotton bud and then delicately dried under airflow (A3 subcategory and B3 subcategory); for another sub-category G-Coat was laced through a micro-tip dispenser and light treated for twenty seconds (A2 subcategory and B2 subcategory); the rest 20 specimens were left without any coating (A1 subcategory and B1 subcategory). It was observed that in subcategory A1 and A3 there was continuous decline in emission of fluoride ion as the days progressed. However there was an increase in emission of fluoride in A2 subcategory on moving to day 5 from day 1. However, from day 5 onwards decline in fluoride emission was observed in A2 subcategory. It was concluded that both materials studied (GIC and Zirconomer) exhibited fluoride emission whether or not they were surface-coated for protection.

Comparative analysis of secondary metabolite gene clusters in different strains of Magnaporthe oryzae.

Rice blast caused by Magnaporthe oryzae continues to be a major constraint in rice production worldwide. Rice is one of the staple crops in India and rice blast causes huge economic losses. Interestingly, the Indian subcontinent is the centre for origin and diversity of rice as well as the Magnaporthe species complex. Secondary metabolites are known to play important role in pathogenesis and M. oryzae has high potential of genes involved in secondary metabolism but, unfortunately most of them remain uncharacterized. In the present study, we analysed the draft genome assemblies of M. oryzae strains isolated from different parts of India, for putative secondary metabolite key gene (SMKG) clusters encoding polyketide synthases, non-ribosomal peptide synthetases, diterpene cyclases and dimethylallyl tryptophan synthase. Based on the complete genome sequence of 70-15 strain and its previous reports of identified SMKGs, we have identified the key genes for the interrogated strains. Expression analysis of these genes amongst different strains indicates how they have evolved depending on the host and environmental conditions. To our knowledge, this study is first of its kind where the secondary metabolism genes and their role in functional adaptation were studied across several strains of M. oryzae.

MoLAEA Regulates Secondary Metabolism in Magnaporthe oryzae.

Fungi are rich sources of secondary metabolites of pharmaceutical importance, such as antibiotics, antitumor agents, and immunosuppressants, as well as of harmful toxins. Secondary metabolites play important roles in the development and pathogenesis of fungi. LaeA is a global regulator of secondary metabolism and was originally reported in Aspergillus nidulans; however, its role in secondary metabolism in Magnaporthe oryzae has not yet been reported. Here, we investigated the role of a gene homologous to LAEA (loss of AflR expression) of Aspergillus spp. in Magnaporthe oryzae, named M. oryzaeLAEA (MoLAEA). Studies on MoLAEA overexpression and knockdown strains have suggested that this gene acts as a negative regulator of sporulation and melanin synthesis. However, it is not involved in the growth and pathogenesis of M. oryzae Transcriptomic data indicated that MoLAEA regulated genes involved in secondary metabolism. Interestingly, we observed (for the first time, to our knowledge) that this gene is involved in benzylpenicillin (penicillin G) synthesis in M. oryzae Overexpression of MoLAEA increased penicillin G production, whereas the silenced strain showed a complete absence of penicillin G compared to its presence in the wild type. We also observed that MoLaeA interacted with MoVeA, a velvet family protein involved in fungal development and secondary metabolism, in the nucleus. This study showed that though MoLAEA may not make any contribution in rice blast fungal pathogenesis, it regulates secondary metabolism in M. oryzae and thus can be further studied for identifying other new uncharacterized metabolites in this fungus.IMPORTANCEM. oryzae causes blast disease, the most serious disease of cultivated rice affecting global rice production. The genome of M. oryzae has been shown to have a number of genes involved in secondary metabolism, but most of them are uncharacterized. In fact, compared to studies of other filamentous fungi, hardly any work has been done on secondary metabolism in M. oryzae It is shown here (for the first time, to our knowledge) that penicillin G is being synthesized in M. oryzae and that MoLAEA is involved in this process. This is the first step in understanding the penicillin G biosynthesis pathway in M. oryzae This study also unraveled the details of how MoLaeA works by forming a nuclear complex with MoVeA in M. oryzae, thus indicating functional conservation of such a gene across filamentous fungi. All these findings open up avenues for more relevant investigations on the genetic regulation of secondary metabolism in M. oryzae.