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Introduction: Plant–microbe interactions play a vital role in the development of strategies to manage pathogen-induced destructive diseases that cause enormous crop losses every year. Rice blast is one of the severe diseases to rice Oryza sativa (O. sativa) due to Magnaporthe grisea (M. grisea) fungus. Protein–protein interaction (PPI) between rice and fungus plays a key role in causing rice blast disease. Methods: In this paper, four genomic information-based models such as (i) the interolog, (ii) the domain, (iii) the gene ontology, and (iv) the phylogenetic-based model are developed for predicting the interaction between O. sativa and M. grisea in a whole-genome scale. Results and Discussion: A total of 59,430 interacting pairs between 1,801 rice proteins and 135 blast fungus proteins are obtained from the four models. Furthermore, a machine learning model is developed to assess the predicted interactions. Using composition-based amino acid composition (AAC) and conjoint triad (CT) features, an accuracy of 88% and 89% is achieved, respectively. When tested on the experimental dataset, the CT feature provides the highest accuracy of 95%. Furthermore, the specificity of the model is verified with other pathogen–host datasets where less accuracy is obtained, which confirmed that the model is specific to O. sativa and M. grisea. Understanding the molecular processes behind rice resistance to blast fungus begins with the identification of PPIs, and these predicted PPIs will be useful for drug design in the plant science community. [ABSTRACT FROM AUTHOR]

IntroductionPlant–microbe interactions play a vital role in the development of strategies to manage pathogen-induced destructive diseases that cause enormous crop losses every year. Rice blast is one of the severe diseases to rice Oryza sativa (O. sativa) due to Magnaporthe grisea (M. grisea) fungus. Protein–protein interaction (PPI) between rice and fungus plays a key role in causing rice blast disease.MethodsIn this paper, four genomic information-based models such as (i) the interolog, (ii) the domain, (iii) the gene ontology, and (iv) the phylogenetic-based model are developed for predicting the interaction between O. sativa and M. grisea in a whole-genome scale.Results and DiscussionA total of 59,430 interacting pairs between 1,801 rice proteins and 135 blast fungus proteins are obtained from the four models. Furthermore, a machine learning model is developed to assess the predicted interactions. Using composition-based amino acid composition (AAC) and conjoint triad (CT) features, an accuracy of 88% and 89% is achieved, respectively. When tested on the experimental dataset, the CT feature provides the highest accuracy of 95%. Furthermore, the specificity of the model is verified with other pathogen–host datasets where less accuracy is obtained, which confirmed that the model is specific to O. sativa and M. grisea. Understanding the molecular processes behind rice resistance to blast fungus begins with the identification of PPIs, and these predicted PPIs will be useful for drug design in the plant science community.

Mediator complex is a multiprotein complex that regulates RNA polymerase II-mediated transcription. Moreover, it functions in several signaling pathways, including those involved in response to biotic and abiotic stresses. We used virus-induced gene silencing (VIGS) to study the functions of two genes, namely OsMED16 and OsMED25 in response to biotic and abiotic stresses in rice. Both genes were differentially induced by Magnaporthe grisea (M. grisea), the causative agent of blast disease, hormone treatment, and abiotic stress. We found that both BMV: OsMED16- and BMV: OsMED25-infiltrated seedlings reduced the resistance to M. grisea by regulating the accumulation of H2O2 and expression of defense-related genes. Furthermore, BMV: OsMED16-infiltrated seedlings decreased the tolerance to cold by increasing the malondialdehyde (MDA) content and reducing the expression of cold-responsive genes.

Mediator complex is a multiprotein complex that regulates RNA polymerase II-mediated transcription. Moreover, it functions in several signaling pathways, including those involved in response to biotic and abiotic stresses. We used virus-induced gene silencing (VIGS) to study the functions of two genes, namely OsMED16 and OsMED25 in response to biotic and abiotic stresses in rice. Both genes were differentially induced by Magnaporthe grisea (M. grisea), the causative agent of blast disease, hormone treatment, and abiotic stress. We found that both BMV: OsMED16- and BMV: OsMED25-infiltrated seedlings reduced the resistance to M. grisea by regulating the accumulation of H2O2 and expression of defense-related genes. Furthermore, BMV: OsMED16-infiltrated seedlings decreased the tolerance to cold by increasing the malondialdehyde (MDA) content and reducing the expression of cold-responsive genes. [ABSTRACT FROM AUTHOR]

Magnaporthe oryzae infects rice, wheat, and many grass species in the Poaceae family by secreting protein effectors. Here, we analyzed the distribution, sequence variation, and genomic context of effector candidate (EFC) genes in 31 isolates that represent five pathotypes of M. oryzae, three isolates of M. grisea, a sister species of M. oryzae, and one strain each for eight species in the family Magnaporthaceae to investigate how the host range expansion of M. oryzae has likely affected the evolution of effectors. We used the EFC genes of M. oryzae strain 70-15, whose genome has served as a reference for many comparative genomics analyses, to identify their homologs in these strains. We also analyzed the previously characterized avirulence (AVR) genes and single-copy orthologous (SCO) genes in these strains, which showed that the EFC and AVR genes evolved faster than the SCO genes. The EFC and AVR repertoires among M. oryzae pathotypes varied widely probably because adaptation to individual hosts exerted different types of selection pressure. Repetitive DNA elements appeared to have caused the variation of some EFC genes. Lastly, we analyzed expression patterns of the AVR and EFC genes to test the hypothesis that such genes are preferentially expressed during host infection. This comprehensive dataset serves as a foundation for future studies on the genetic basis of the evolution and host specialization in M. oryzae.

Magnaporthe oryzae infects rice, wheat, and many grass species in the Poaceae family by secreting protein effectors. Here, we analyzed the distribution, sequence variation, and genomic context of effector candidate (EFC) genes in 31 isolates that represent five pathotypes of M. oryzae , three isolates of M. grisea , a sister species of M. oryzae , and one strain each for eight species in the family Magnaporthaceae to investigate how the host range expansion of M. oryzae has likely affected the evolution of effectors. We used the EFC genes of M. oryzae strain 70-15, whose genome has served as a reference for many comparative genomics analyses, to identify their homologs in these strains. We also analyzed the previously characterized avirulence (AVR) genes and single-copy orthologous (SCO) genes in these strains, which showed that the EFC and AVR genes evolved faster than the SCO genes. The EFC and AVR repertoires among M. oryzae pathotypes varied widely probably because adaptation to individual hosts exerted different types of selection pressure. Repetitive DNA elements appeared to have caused the variation of some EFC genes. Lastly, we analyzed expression patterns of the AVR and EFC genes to test the hypothesis that such genes are preferentially expressed during host infection. This comprehensive dataset serves as a foundation for future studies on the genetic basis of the evolution and host specialization in M. oryzae. [ABSTRACT FROM AUTHOR]

Blast disease caused by Magnaporthe species is a major threat to the productivity of agriculturally important cereal crops including rice, pearl millet, wheat, oats, barley and minor millets etc. Two major species of Magnaporthe , viz. M. oryzae and M. grisea cause approximately 10–30% annual yield loss in their respective host crop. Imparting long-term resistance in the host necessitates the detailed knowledge on the molecular basis of pathogen virulence factors. Thus, here we have made an attempt to characterize the virulence factors (VFs) from M. grisea strain PMg-Dl in comparison to M. oryzae strain RMg-Dl. The functional gene ontology annotation revealed predominance of membrane system (GO: 0016020, GO:0016021), ion-binding system (GO:0043167; metal binding GO:0046872; zinc ion binding GO:0008270) etc. Further, in silico domain analysis of eight uncharacterized preferential virulence factor genes, viz. MBSD02000619.1.g3349 , MBSD02000495.1.G5524 , and MBSD02000009.1.G8852 and Mg.00g031250.m01, Mg.00g080210.m01, Mg.00g080580.m01, Mg.00g034390.m01 and Mg.00g016130.m01 showed the target genes belongingness to MFS family, integrase, protease, transcription factors etc. The PCR analysis showed the presence of eight target VFs in the Magnaporthe isolates infecting rice, pearl millet, finger millet and other grasses, respectively. The functional analysis of uncharacterized target preferential virulence factor genes (M. oryzae :3; M. grisea: 5) in three host systems viz., rice, pearl millet and finger millet at eight different time points confirmed their role as virulence factors in Magnaporthe system. The results are useful in further dissecting the Magnaporthe -host interactions by gene knock-out and knock in approaches through transgenics and genome editing to develop resistant cultivars soon. • Eight potential uncharacterized virulence factors (VFs) were identified from Magnaporthe species. • Each VFs was a part of different protein family including MFS, integrase, receptor etc. • The functional analysis of hypothetical genes confirmed their role as virulence factors in Magnaporthe system. • VFs genes plays a key role in modulating the blast susceptibility in rice, pearl millet and finger millet hosts. [ABSTRACT FROM AUTHOR]

Eucalyptus oil possesses a wide spectrum of biological activity, including anti-microbial, fungicidal, herbicidal, acaricidal and nematicidal properties. We studied anti-fungal activities of the leaf oil extracted from Eucalyptus. grandis × E. urophylla. Eleven plant pathogenic fungi were tested based on the mycelium growth rates with negative control. The results showed that Eucalyptus oil has broad-spectrum inhibitory effects toward these fungi. Remarkable morphological and structural alterations of hypha have been observed for Magnaporthe grisea after the treatment. The mRNA genome array ofM. grisea was used to detect genes that were differentially expressed in the test strains treated by the Eucalyptus oil than the normal strains. The results showed 1919 genes were significantly affected, among which 1109 were down-regulated and 810 were up-regulated (p < 0.05, absolute fold change >2). According to gene ontology annotation analysis, these differentially expressed genes may cause abnormal structures and physiological function disorders, which may reduce the fungus growth. These results show the oil has potential for use in the biological control of plant disease as a green biopesticide. [ABSTRACT FROM AUTHOR]

Eucalyptus oil possesses a wide spectrum of biological activity, including anti-microbial, fungicidal, herbicidal, acaricidal and nematicidal properties. We studied anti-fungal activities of the leaf oil extracted from Eucalyptus. grandis × E. urophylla. Eleven plant pathogenic fungi were tested based on the mycelium growth rates with negative control. The results showed that Eucalyptus oil has broad-spectrum inhibitory effects toward these fungi. Remarkable morphological and structural alterations of hypha have been observed for Magnaporthe grisea after the treatment. The mRNA genome array of M. grisea was used to detect genes that were differentially expressed in the test strains treated by the Eucalyptus oil than the normal strains. The results showed 1919 genes were significantly affected, among which 1109 were down-regulated and 810 were up-regulated (p < 0.05, absolute fold change >2). According to gene ontology annotation analysis, these differentially expressed genes may cause abnormal structures and physiological function disorders, which may reduce the fungus growth. These results show the oil has potential for use in the biological control of plant disease as a green biopesticide.

The target site of tolprocarb, a novel systemic fungicide used for controlling rice blast, was investigated. Tolprocarb decolorized the mycelia of Magnaporthe grisea; the decolorization was reversed by adding scytalone or 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN). This result suggested that the target site of tolprocarb was polyketide synthase (PKS), which regulated polyketide synthesis and pentaketide cyclization in melanin biosynthesis. Further, we produced a transgenic Aspergillus oryzae, which possessed the PKS gene of M. grisea, and performed in vitro assays of PKS using membrane fractions from the transgenic A. oryzae. Compared with some conventional melanin biosynthesis inhibitors (cMBIs), tolprocarb only inhibited PKS activity in vitro. These results indicated that tolprocarb's target protein in M. grisea was PKS, which differentiates this fungicide from other cMBIs. [ABSTRACT FROM AUTHOR]

Abstract: Short antimicrobial peptides represent an alternative to fight pathogen infections. PAF26 is a hexapeptide identified previously by a combinatorial approach against the fungus Penicillium digitatum and shows antimicrobial properties towards certain phytopathogenic fungi. In this work, PAF26 was used as lead compound and its properties were compared with two series of derivatives, obtained by either systematic alanine substitution or N-terminal amino acid addition. The alanine scan approach underlined the optimized sequence of PAF26 in terms of potency and permeation capability, and also the higher contribution of the cationic residues to these properties. The N-terminal addition of amino acids resulted in new heptapeptides with variations in their antimicrobial characteristics, and very low cytolysis to human red blood cells. Positive (Arg or Lys) and aromatic (Phe or Trp) residue addition increased broad spectrum activity of PAF26. Noteworthy, addition of selected residues had specific effects on the properties of derivatives of PAF26. [Copyright &y& Elsevier]

Abstract: In order to understand the defense machinery in the model cereal crop rice, we performed a large-scale analysis of rice gene expression in response to rice blast Magnaporthe grisea (M. grisea) or Magnaporthe oryzae and bacterial blight Xanthomonas oryzae pv. oryzae (Xoo) during the early incompatible and compatible interactions. Using a gene chip containing 10254 rice cDNAs representing 9240 unique genes, we identified 794 and 612 genes differentially expressed in the incompatible and compatible rice–M. grisea interactions, respectively, with 274 genes co-regulated during both interactions. In the rice–Xoo pathosystem, 454 and 498 differentially expressed genes were identified in the incompatible and compatible interactions, respectively, including 237 co-regulated genes in the both interactions. By clustering differentially regulated genes from all these interactions, we identified 29 co-regulated genes in the all four interactions, and 86 and 74 co-regulated genes in the two incompatible and two compatible interactions, respectively. These differentially expressed genes could be classified into three categories, including M. grisea- and Xoo-regulated, M. grisea-specific, and Xoo-specific. The expression patterns of representative defense-related genes were further confirmed by RT-PCR. The large-scale expression data from our microarray analysis indicated the existence of distinctive as well as shared defense pathways between the rice–M. grisea and rice–Xoo interactions. [Copyright &y& Elsevier]

The induction of host-defense in plants is a reliable approach for the management of diseases in crop plants. In the present study, biotic agents like Pseudomonas fluorescens, Trichoderma virens and neem leaf extract have been evaluated for their efficacy to induce resistance against blast disease of pearl millet caused by Magnaporthe grisea and promote growth. Seeds treated with P. fluorescens exhibited maximum seed germination of 83.50% and seedling vigor of 1447.93 followed by seed treatment with T. virens and neem leaf extract. Seeds treated with P. fluorescens showed improved vegetative and reproductive parameters compared with other biotic agents. Under greenhouse conditions, P. fluorescens offered maximum disease protection of 62.25% against blast disease followed by T. virens with 58.25%. It was also found that the activity of the defense-related enzymes like Phenylalanine ammonia lyase (PAL), Peroxidase (POX), Lipoxygenase (LOX) and β-1,3-glucanase was increased considerably in P. fluorescens treated and challenge inoculated seedlings thereby indicating that these defense enzymes play a pivotal role in induction of blast disease resistance in pearl millet. • Seed priming with biotic agents (P. fluorescens , T. virens and neem leaf extract) enhanced seed and plant growth parameters in pearl millet. • The biotic agents also induced resistance in pearl millet against M. grisea the causal agent of blast disease. • The disease protection studies were validated with increased activity of defense-related enzymes (PAL, POX, LOX and β-1,3-glucanse). [ABSTRACT FROM AUTHOR]