A full-length enriched cDNA library and expressed sequence tag analysis of the parasitic weed, Striga hermonthica

<p>Abstract</p> <p>Background</p> <p>The obligate parasitic plant witchweed (<it>Striga hermonthica</it>) infects major cereal crops such as sorghum, maize, and millet, and is the most devastating weed pest in Africa. An understanding of the nature of its parasitism would contribute to the development of more sophisticated management methods. However, the molecular and genomic resources currently available for the study of <it>S. hermonthica </it>are limited.</p> <p>Results</p> <p>We constructed a full-length enriched cDNA library of <it>S. hermonthica</it>, sequenced 37,710 clones from the library, and obtained 67,814 expressed sequence tag (EST) sequences. The ESTs were assembled into 17,317 unigenes that included 10,319 contigs and 6,818 singletons. The <it>S. hermonthica </it>unigene dataset was subjected to a comparative analysis with other plant genomes or ESTs. Approximately 80% of the unigenes have homologs in other dicotyledonous plants including <it>Arabidopsis</it>, poplar, and grape. We found that 589 unigenes are conserved in the hemiparasitic <it>Triphysaria </it>species but not in other plant species. These are good candidates for genes specifically involved in plant parasitism. Furthermore, we found 1,445 putative simple sequence repeats (SSRs) in the <it>S. hermonthica </it>unigene dataset. We tested 64 pairs of PCR primers flanking the SSRs to develop genetic markers for the detection of polymorphisms. Most primer sets amplified polymorphicbands from individual plants collected at a single location, indicating high genetic diversity in <it>S. hermonthica</it>. We selected 10 primer pairs to analyze <it>S. hermonthica </it>harvested in the field from different host species and geographic locations. A clustering analysis suggests that genetic distances are not correlated with host specificity.</p> <p>Conclusions</p> <p>Our data provide the first extensive set of molecular resources for studying <it>S. hermonthica</it>, and include EST sequences, a comparative analysis with other plant genomes, and useful genetic markers. All the data are stored in a web-based database and freely available. These resources will be useful for genome annotation, gene discovery, functional analysis, molecular breeding, epidemiological studies, and studies of plant evolution.</p

Anti-rat myoglobin antisera in the immunocytochemical diagnosis of rhabdomyosarcomas of rats. Vet

Abstract. Anti-rat myoglobin (Mb) was prepared and used in the avidin-biotin-peroxidase complex (ABC) method on paraffin-embedded sections of nine soft tissue tumors (including two rhabdomyosarcomas) of rats. Distribution and nature of the reactive substance to Mb antiserum were compared to those of desmin antiserum. Rat Mb was isolated from the skeletal muscle; monospecificity of the rat antiserum was confirmed by the immunoblotting procedures. The Mb antiserum reacted specifically to normal and neoplastic striated muscle cells. Mb-staining reactions were present diffusely in the cytoplasm, while desmin-staining substances were localized at Z-bands or were diffuse in the cytoplasm as separated aggregates. Reaction to the Mb serum was also detected in cells of thick portions of Henle&apos;s loop and distal convoluted tubules

Arabidopsis thaliana GYRB3 Does Not Encode a DNA Gyrase Subunit

, is predicted to encode four gyrase subunits: AtGyrA, AtGyrB1, AtGyrB2 and AtGyrB3. temperature-sensitive strain, whereas AtGyrB2 can. Yeast two-hybrid analysis suggests that AtGyrB3 cannot bind to AtGyrA or form a dimer.These data strongly suggest that AtGyrB3 is not a gyrase subunit but has another unknown function. One possibility is that it is a nuclear protein with a role in meiosis in pollen

SRFR1 Negatively Regulates Plant NB-LRR Resistance Protein Accumulation to Prevent Autoimmunity

Plant defense responses need to be tightly regulated to prevent auto-immunity, which is detrimental to growth and development. To identify negative regulators of Resistance (R) protein-mediated resistance, we screened for mutants with constitutive defense responses in the npr1-1 background. Map-based cloning revealed that one of the mutant genes encodes a conserved TPR domain-containing protein previously known as SRFR1 (SUPPRESSOR OF rps4-RLD). The constitutive defense responses in the srfr1 mutants in Col-0 background are suppressed by mutations in SNC1, which encodes a TIR-NB-LRR (Toll Interleukin1 Receptor-Nucleotide Binding-Leu-Rich Repeat) R protein. Yeast two-hybrid screens identified SGT1a and SGT1b as interacting proteins of SRFR1. The interactions between SGT1 and SRFR1 were further confirmed by co-immunoprecipitation analysis. In srfr1 mutants, levels of multiple NB-LRR R proteins including SNC1, RPS2 and RPS4 are increased. Increased accumulation of SNC1 is also observed in the sgt1b mutant. Our data suggest that SRFR1 functions together with SGT1 to negatively regulate R protein accumulation, which is required for preventing auto-activation of plant immunity