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P C Ronald

Publications and source records attributed to P C Ronald.

26 records · Page 2Linked to original sources
PubMed1996

The cloned gene, Xa21, confers resistance to multiple Xanthomonas oryzae pv. oryzae isolates in transgenic plants.

The cloned rice gene, Xa21, confers resistance to multiple pathogen isolates of Xanthomonas oryzae pv. oryzae in transgenic plants. The resistance phenotype was stably transmitted to T1 progeny and inherited as a single locus. The T1 progeny were tested for resistance to 32 X. oryzae pv. oryzae isolates from eight countries. Both the engineered line and the donor line showed resistance to 29 isolates and susceptibility to three isolates. The identical resistance spectrum of both lines indicates that the presence of a single member of a multigene family, Xa21, is sufficient to confer multi-isolate resistance. The results presented here have important implications for engineering disease resistance in crop plants.

Base Sequence↗
PubMed1995

Metaphase and interphase fluorescence in situ hybridization mapping of the rice genome with bacterial artificial chromosomes.

Fluorescence in situ hybridization (FISH) is a powerful tool for physical mapping in human and other mammalian species. However, application of the FISH technique has been limited in plant species, especially for mapping single- or low-copy DNA sequences, due to inconsistent signal production in plant chromosome preparations. Here we demonstrate that bacterial artificial chromosome (BAC) clones can be mapped readily on rice (Oryza sativa L.) chromosomes by FISH. Repetitive DNA sequences in BAC clones can be suppressed efficiently by using rice genomic DNA as a competitor in the hybridization mixture. BAC clones as small as 40 kb were successfully mapped. To demonstrate the application of the FISH technique in physical mapping of plant genomes, both anonymous BAC clones and clones closely linked to a rice bacterial blight-resistance locus, Xa21, were chosen for analysis. The physical location of Xa21 and the relationships among the linked clones were established, thus demonstrating the utility of FISH in plant genome analysis.

Animals↗
PubMed1995

Construction of a rice bacterial artificial chromosome library and identification of clones linked to the Xa-21 disease resistance locus.

A bacterial artificial chromosome (BAC) library consisting of 11,000 clones with an average DNA insert size of 125 kb was constructed from rice nuclear DNA. The BAC clones were stable in E. coli after 100 generations of serial growth. Transformation of the BAC clones by electroporation into E. coli was highly efficient and increased with decreasing size of the DNA inserts. The library was evaluated for the presence of organellar, repeated, and telomeric sequences. A very low percentage (< 0.3%) of the library consisted of chloroplast and mitochondrial clones. Eighteen BACs were identified that hybridized with an Arabidopsis telomere repeat. Sixteen BACs hybridized with the AA genome-specific repetitive sequence pOs48. Twelve clones were isolated that hybridized with three DNA markers linked to the Xa-21 disease resistance locus. The results indicate that the BAC system can be used to clone and manipulate large pieces of plant DNA efficiently.

Arabidopsis↗
PubMed1994

Saturated molecular map of the rice genome based on an interspecific backcross population.

A molecular map has been constructed for the rice genome comprised of 726 markers (mainly restriction fragment length polymorphisms; RFLPs). The mapping population was derived from a backcross between cultivated rice, Oryza sativa, and its wild African relative, Oryza longistaminata. The very high level of polymorphism between these species, combined with the use of polymerase chain reaction-amplified cDNA libraries, contributed to mapping efficiency. A subset of the probes used in this study was previously used to construct an RFLP map derived from an inter subspecific cross, providing a basis for comparison of the two maps and of the relative mapping efficiencies in the two crosses. In addition to the previously described PstI genomic rice library, three cDNA libraries from rice (Oryza), oat (Avena) and barley (Hordeum) were used in this mapping project. Levels of polymorphism detected by each and the frequency of identifying heterologous sequences for use in rice mapping are discussed. Though strong reproductive barriers isolate O. sativa from O. longistaminata, the percentage of markers showing distorted segregation in this backcross population was not significantly different than that observed in an intraspecific F2 population previously used for mapping. The map contains 1491 cM with an average interval size of 4.0 cM on the framework map, and 2.0 cM overall. A total of 238 markers from the previously described PstI genomic rice library, 250 markers from a cDNA library of rice (Oryza), 112 cDNA markers from oat (Avena), and 20 cDNA markers from a barley (Hordeum) library, two genomic clones from maize (Zea), 11 microsatellite markers, three telomere markers, eleven isozymes, 26 cloned genes, six RAPD, and 47 mutant phenotypes were used in this mapping project. Applications of a molecular map for plant improvement are discussed.

Avena↗
PubMed1992

Genetic and physical analysis of the rice bacterial blight disease resistance locus, Xa21.

Nearly isogenic lines (NILs) of rice (Oryza sativa) differing at a locus conferring resistance to the pathogen Xanthomonas oryzae pv. oryzae were surveyed with 123 DNA markers and 985 random primers using restriction fragment length plymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis. One chromosome 11 marker (RG103) detected polymorphism between the NILs that cosegregated with Xa21. All other chromosome 11 DNA markers tested were monomorphic between the NILs, localizing the Xa21 introgressed region to an 8.3 cM interval on chromosome 11. Furthermore, we identified two polymerase chain reaction (PCR) products (RAPD2148 and RAPD818) that detected polymorphisms between the NILs. Genomic sequences hybridizing with RAPD818, RAPD248 and RG103 were duplicated specifically in the Xa21 NIL. All three markers cosegregated with the resistance locus, Xa21, in a F2 population of 386 progeny. Based on the frequency with which we recovered polymorphic Xa21-linked markers, we estimated the physical size of the introgressed region to be approximately 800 kb. This estimation was supported by physical mapping (using pulsed field gel electrophoresis) of the sequences hybridizing with the three Xa21-linked DNA markers. The results showed that the three Xa21-linked markers are physically close to each other, with one copy of the RAPD818 sequences located within 60 kb of RAPD248 and the other copy within 270 kb of RG103. None of the enzymes tested generated a DNA fragment that hybridized with all three of the markers indicating that the introgressed region containing the resistance locus Xa21 is probably larger than 270 kb.

Chromosome Mapping↗
PubMed1992

The cloned avirulence gene avrPto induces disease resistance in tomato cultivars containing the Pto resistance gene.

Resistance of tomato plants to the bacterial pathogen Pseudomonas syringae pv. tomato race 0 is controlled by the locus Pto. A bacterial avirulence gene was cloned by constructing a cosmid library from an avirulent P. syringae pv. tomato race, conjugating the recombinants into a strain of P. syringae pv. maculicola virulent on a tomato cultivar containing Pto, and screening for those clones that converted the normally virulent phenotype to avirulence. The cloned gene, designated avrPto, reduced multiplication of P. syringae pv. tomato transconjugants specifically on Pto tomato lines, as demonstrated by bacterial growth curve analyses. Analysis of F2 populations revealed cosegregation of resistance to P. syringae pv. tomato transconjugants carrying avrPto with resistance to P. syringae pv. tomato race 0. Surprisingly, mutation of avrPto in P. syringae pv. tomato race 0 does not eliminate the avirulent phenotype of race 0, suggesting that additional, as yet uncharacterized, avirulence genes and/or resistance genes may contribute to specificity in the avrPto-Pto interaction. Genetic analysis indicates that this resistance gene(s) would be tightly linked to Pto. Interestingly, P. syringae pv. glycinea transconjugants carrying avrPto elicit a typical hypersensitive resistant response in the soybean cultivar Centennial, suggesting conservation of Pto function between two crop plants, tomato and soybean.

Cell Division↗
PubMedDate not supplied

The avirulence gene avrBs1 from Xanthomonas campestris pv. vesicatoria encodes a 50-kD protein.

A gene cloned from Xanthomonas campestris pv. vesicatoria race 2, avrBs1, specified avirulence on pepper cultivars containing the resistance gene Bs1. A series of exonuclease III deletions were made on a 3.2-kbp DNA fragment that determined full avirulence activity, observed as hypersensitive response (HR) induction. The deletion products were subcloned into the broad host range cloning vector pLAFR3, conjugated into a virulent X. c. pv. vesicatoria race 1 strain, 82-8, and scored for their ability to induce a HR on a pepper cultivar (ECW10R) containing the resistance gene Bs1. A span of approximately 1.8 kbp of DNA was necessary for full induction of the HR. The nucleotide sequence revealed two open reading frames (ORFs) capable of encoding proteins of 12.3 and 49.8 kD, designated ORF1 and ORF2, respectively. Deletions into ORF1 altered the HR-inducing activity to give an intermediate phenotype. Deletions into ORF2 completely destroyed activity. When the ORF2 coding region was driven by the lacZ promoter on plasmid pLAFR3 (placD), full avirulence activity was restored, indicating that ORF2 alone can induce the HR. Antisera raised to a beta-galactosidase-ORF2 fusion protein reacted with a 50-kD protein in X. c. pv. vesicatoria race 1 (placD) transconjugants. The deduced amino acid sequence of ORF2 had approximately 47% overall homology to the carboxyl terminus of the avirulence gene, avrA, isolated from Pseudomonas syringae pv. glycinea race 6, and 86% homology over a region of 49 amino acids. P. s. pv. glycinea, however, did not induce an HR on ECW10R plants.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗