Genome-wide identification, evolutionary and expression analyses of putative Fe-S biogenesis genes in rice (Oryza sativa).

Iron-sulfur (Fe-S) proteins are ubiquitous in nature and carry Fe-S clusters (ISCs) as prosthetic groups that are essential in maintaining basic biological processes such as photosynthesis, respiration, nitrogen fixation, and DNA repair. In the present investigation, a comprehensive genome-wide analysis was carried out to find all the genes involved in the formation of ISCs in rice ( Oryza sativa L.) through a systematic EST and genomic DNA sequence data mining. This analysis profiled 44 rice ISC genes (OsISCs) that were identified using in silico analysis. Multiple sequence alignment and phylogenetic analysis revealed that these genes were highly conserved among bacteria, fungi, animals, and plants. EST analysis and RT-PCR assays demonstrated that all OsISCs were active and that the transcript abundance of some OsISCs was tissue specific. The results of this study will assist further investigations to identify and elucidate the structural components involved in the assembly, biogenesis, and regulation of OsISCs. Thus, the outcome of the present study provides basic genomic information for the OsISC and will pave the way for elucidating the precise role of OsISCs in plant growth and development in the future. Also, it may enable us in the future to enhance the crop yield, uptake of Fe, and protection against abiotic and biotic stress.

Targeted spatio-temporal expression based characterization of state of infection and time-point of maximum defense in wheat NILs during leaf rust infection.

Leaf rust, caused by the fungus Puccinia triticina, is the most devastating disease of wheat worldwide, which sometimes becomes epidemic. The pathogen evolves into new strains, making its control difficult. Though more than 60 leaf rust resistant genes are now known, only limited insight is available into the molecular mechanism involved in this host pathogen interaction. In the present study, quantitative real-time PCR based differential gene expression profiling was examined for five target genes encoding for chitinase3, ß-1,3/1,4 glucanase, thaumatin-like protein, peroxidase2 and mitogen activated protein kinase1 to unravel their coordinated action during compatible and incompatible interaction, to inhibit the pathogen progression and to identify the time-period of maximum defense activity. Spatio-temporal expression profiling suggested that the maximum defense activity occurred at 12-24 hours post inoculation, whereas the state of infection and degree of resistance was predicted using coordinated unique expression signatures of target genes. The significant differences of targeted gene expression between resistant mock inoculated, resistant infected and susceptible infected plants were evaluated using t test at significance level of p < 0.01. The differences occurred can be attributed to the presence of seedling leaf rust resistance Lr28 gene, which facilitated prevention of leaf rust infection in resistant wheat plants.

Analysis of differentially expressed genes in leaf rust infected bread wheat involving seedling resistance gene Lr28.

Genome-wide transcriptome analysis of seedling resistance to leaf rust conferred by Lr28 gene in wheat (Triticum aestivum L.) was conducted to identify differentially expressed genes during incompatible interaction. A virulent leaf rust race 77-5 was used for inoculation of resistant (HD2329+Lr28) and susceptible (HD2329 - Lr28) wheat NILs and cDNA-AFLP analyses was carried out. As many as 223 differential transcripts appeared following leaf rust inoculation; these included 122 transcripts that appeared exclusively in resistant NIL, whereas 39 transcripts appeared both in resistant and susceptible NILs. Sequence analyses of 37 transcripts, which appeared in the resistant NIL revealed that 15 transcripts had homology with genes involved in protein synthesis, signal transduction, transport, disease resistance and metabolism. The functions of remaining 22 transcripts could not be determined; these included six novel genes reported for the first time in wheat. Specific primers could be designed for 18 of the 37 transcripts, which included genes with putative and unknown functions. Quantitative real time PCR analysis was conducted using these 18 pairs of primers. A majority (13) of these transcripts appeared within 48h reaching a peak value at 96h in resistant NIL signifying their role in providing leaf rust resistance.