Proteomic and genetic approaches to identifying defence-related proteins in rice challenged with the fungal pathogen Rhizoctonia solani

Sheath blight, caused by the fungus Rhizoctonia solani, is a major disease of rice world-wide, but little is known about the host response to infection. The objective of this study was to identify proteins and DNA markers in resistant and susceptible rice associated with response to infection by R. solani. Replicated two-dimensional polyacrylamide gel electrophoresis experiments were conducted to detect proteins differentially expressed under inoculated and non-inoculated conditions. Tandem mass spectra analysis using electrospray ionization quadrupole-time of flight mass spectrometry (ESI Q-TOF MS) was carried out for protein identification with the NCBI non-redundant protein database. Seven proteins were increased after inoculation in both susceptible and resistant plants. Six of the seven proteins were identified with presumed antifungal, photosynthetic and proteolytic activities. An additional 14 proteins were detected in the response of the resistant line. Eleven of the 14 proteins were identified with presumed functions relating to antifungal activity, signal transduction, energy metabolism, photosynthesis, molecular chaperone, proteolysis and antioxidation. The induction of 3-β- hydroxysteroid dehydrogenase/isomerase was detected for the first time in resistant rice plants after pathogen challenge, suggesting a defensive role of this enzyme in rice against attack by R. solani. The chromosomal locations of four induced proteins were found to be in close physical proximity to genetic markers for sheath blight resistance in two genetic mapping populations. The proteomic and genetic results from this study indicate a complex response of rice to challenge by R. solani that involves simultaneous induction of proteins from multiple defence pathways. © 2006 Blackwell Publishing Ltd

Qualidade sanitária e fisiológica de sementes de abóbora variedade menina Brasileira.

O trabalho teve como objetivos avaliar e correlacionar a qualidade sanitária e fisiológica de sementes de abóbora, variedade Menina Brasileira (Cucurbita moschata.). Foram avaliados dois lotes de sementes de abóbora produzidas no sistema agroecológico e quatro no sistema convencional, com e sem tratamento químico. Os lotes foram submetidos aos testes de sanidade, seguindo a metodologia do “Blotter test”, com congelamento, germinação e vigor (primeira contagem, índice de velocidade de germinação, envelhecimento acelerado e emergência de plântulas). Os resultados indicaram a separação dos lotes de diferentes origens a partir da qualidade sanitária e fisiológica, onde as maiores incidências de fungos foram observadas nos lotes agroecológicos e o maior potencial fisiológico foi observado nos lotes de origem convencional não tratados. Foram encontrados os fungos Fusarium oxysporum, Alternaria alternata, Cladosporium cucumerinum, Aspergillus niger, Penicillium digitatum, Rhizopus stolonifer e Phoma terrestris. A qualidade sanitária não interferiu na qualidade fisiológica das sementes de abóbora, variedade Menina Brasileira

Metabolomic and transcriptomic analysis of the rice response to the bacterial blight pathogen Xanthomonas oryzae pv. oryzae

Bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo), gives rise to devastating crop losses in rice. Disease resistant rice cultivars are the most economical way to combat the disease. The TP309 cultivar is susceptible to infection by Xoo strain PXO99. A transgenic variety, TP309_Xa21, expresses the pattern recognition receptor Xa21, and is resistant. PXO99△raxST, a strain lacking the raxST gene, is able to overcome Xa21-mediated immunity. We used a single extraction solvent to demonstrate comprehensive metabolomics and transcriptomics profiling under sample limited conditions, and analyze the molecular responses of two rice lines challenged with either PXO99 or PXO99△raxST. LC–TOF raw data file filtering resulted in better within group reproducibility of replicate samples for statistical analyses. Accurate mass match compound identification with molecular formula generation (MFG) ranking of 355 masses was achieved with the METLIN database. GC–TOF analysis yielded an additional 441 compounds after BinBase database processing, of which 154 were structurally identified by retention index/MS library matching. Multivariate statistics revealed that the susceptible and resistant genotypes possess distinct profiles. Although few mRNA and metabolite differences were detected in PXO99 challenged TP309 compared to mock, many differential changes occurred in the Xa21-mediated response to PXO99 and PXO99△raxST. Acetophenone, xanthophylls, fatty acids, alkaloids, glutathione, carbohydrate and lipid biosynthetic pathways were affected. Significant transcriptional induction of several pathogenesis related genes in Xa21 challenged strains, as well as differential changes to GAD, PAL, ICL1 and Glutathione-S-transferase transcripts indicated limited correlation with metabolite changes under single time point global profiling conditions

Management of Soil-Borne Diseases of Grain Legumes Through Broad-Spectrum Actinomycetes Having Plant Growth-Promoting and Biocontrol Traits

Chickpea (Cicer arietinum L.) and pigeonpea (Cajanus cajan L.) are the two important grain legumes grown extensively in the semiarid tropics (SAT) of the world, where soils are poor in nutrients and receive inadequate/erratic rainfall. SAT regions are commonly found in Africa, Australia, and South Asia. Chickpea and pigeonpea suffer from about 38 pathogens that cause soil-borne diseases including wilt, collar rot, dry root rot, damping off, stem canker, and Ascochyta/Phytophthora blight, and of which three of them, wilt, collar rot, and dry root rot, are important in SAT regions. Management of these soil-borne diseases are hard, as no one control measure is completely effective. Advanced/delayed sowing date, solarization of soil, and use of fungicides are some of the control measures usually employed for these diseases but with little success. The use of disease-resistant cultivar is the best efficient and economical control measure, but it is not available for most of the soil-borne diseases. Biocontrol of soil-borne plant pathogens has been managed using antagonistic actinobacteria, bacteria, and fungi. Actinobacterial strains of Streptomyces, Amycolatopsis, Micromonospora, Frankia, and Nocardia were reported to exert effective control on soil-borne pathogens and help the host plants to mobilize and acquire macro- and micronutrients. Such novel actinomycetes with wide range of plant growth-promoting (PGP) and antagonistic traits need to be exploited for sustainable agriculture. This chapter gives a comprehensive analysis of important soil-borne diseases of chickpea and pigeonpea and how broad-spectrum actinomycetes, particularly Streptomyces spp., could be exploited for managing them

Comparative genome structure, secondary metabolite, and effector coding capacity across Cochliobolus pathogens.

The genomes of five Cochliobolus heterostrophus strains, two Cochliobolus sativus strains, three additional Cochliobolus species (Cochliobolus victoriae, Cochliobolus carbonum, Cochliobolus miyabeanus), and closely related Setosphaeria turcica were sequenced at the Joint Genome Institute (JGI). The datasets were used to identify SNPs between strains and species, unique genomic regions, core secondary metabolism genes, and small secreted protein (SSP) candidate effector encoding genes with a view towards pinpointing structural elements and gene content associated with specificity of these closely related fungi to different cereal hosts. Whole-genome alignment shows that three to five percent of each genome differs between strains of the same species, while a quarter of each genome differs between species. On average, SNP counts among field isolates of the same C. heterostrophus species are more than 25× higher than those between inbred lines and 50× lower than SNPs between Cochliobolus species. The suites of nonribosomal peptide synthetase (NRPS), polyketide synthase (PKS), and SSP-encoding genes are astoundingly diverse among species but remarkably conserved among isolates of the same species, whether inbred or field strains, except for defining examples that map to unique genomic regions. Functional analysis of several strain-unique PKSs and NRPSs reveal a strong correlation with a role in virulence

Differential Induction of Phenylpropanoid Metabolites in Suspension-Cultured Cells of Sugarcane by Fungal Elicitors

The effect of elicitors isolated from the mycelial walls of Colletotrichum falcatum (the red rot pathogen of sugarcane) and from C. lindemuthianum (a non-pathogen) in suspension-cultured cells of sugarcane was studied. Both the elicitors induced the synthesis of enzymes of the phenyl­propanoid pathway such as PAL, TAL and 4CL and also resulted in the enhanced accumulation of phenolics. However, a specific induction of the defense parameters at higher levels was recorded in suspension cells treated with the pathogen elicitor and no such differential response was observed in the case of the non-pathogen elicitor. Elicitor induced necrosis and browning of cells were observed which suggests an additional evidence that elicitors simulate pathogen infection and thus provide a valuable reason that study on elicitor induced responses may be useful in understanding the host defense mechanisms against the red rot pathogen at molecular level