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G J Lawrence

Publications and source records attributed to G J Lawrence.

At least 19 recordsLinked to original sources
PubMed2005

High-resolution mapping and mutation analysis separate the rust resistance genes Sr31, Lr26 and Yr9 on the short arm of rye chromosome 1.

The stem, leaf and stripe rust resistance genes Sr31, Lr26 and Yr9, located on the short arm of rye chromosome 1, have been widely used in wheat by means of wheat-rye translocation chromosomes. Previous studies have suggested that these resistance specificities are encoded by either closely-linked genes, or by a single gene capable of recognizing all three rust species. To investigate these issues, two 1BL.1RS wheat lines, one with and one without Sr31, Lr26 and Yr9, were used as parents for a high-resolution F2 mapping family. Thirty-six recombinants were identified between two PCR markers 2.3 cM apart that flanked the resistance locus. In one recombinant, Lr26 was separated from Sr31 and Yr9. Mutation studies recovered mutants that separated all three rust resistance genes. Thus, together, the recombination and mutation studies suggest that Sr31, Lr26 and Yr9 are separate closely-linked genes. An additional 16 DNA markers were mapped in this region. Multiple RFLP markers, identified using part of the barley Mla powdery mildew resistance gene as probe, co-segregated with Sr31 and Yr9. One deletion mutant that had lost Sr31, Lr26 and Yr9 retained all Mla markers, suggesting that the family of genes on 1RS identified by the Mla probe does not contain the Sr31, Lr26 or Yr9 genes. The genetic stocks and DNA markers generated from this study should facilitate the future cloning of Sr31, Lr26 and Yr9.

Chromosome Mapping↗
PubMed2005

Development of PCR markers for the selection of wheat stem rust resistance genes Sr24 and Sr26 in diverse wheat germplasm.

The use of major resistance genes is the most cost-effective strategy for preventing stem rust epidemics in Australian wheat crops. The long-term success of this strategy is dependent on combining resistance genes that are effective against all predominant races of the pathogen, a task greatly assisted by the use of molecular markers linked to individual resistance genes. The wheat stem rust resistance genes Sr24 and Sr26 (derived from Agropyron elongatum) and SrR and Sr31 (derived from rye) are available in wheat as segments of alien chromosome translocated to wheat chromosomes. Each of these genes provides resistance to all races of wheat stem rust currently found in Australia . We have developed robust PCR markers for Sr24 and Sr26 (this study) and SrR and Sr31 (previously reported) that are applicable across a wide selection of Australian wheat germplasm. Wheat lines have recently become available in which the size of the alien segments containing Sr26, SrR and Sr31 has been reduced. Newly developed PCR-markers can be used to identify the presence of the shorter alien segment in all cases. Assuming that these genes have different gene-for-gene specificities and that the wheat industry will discourage the use of varieties carrying single genes only, the newly developed PCR markers will facilitate the incorporation of two or more of the genes Sr24, Sr26, SrR and Sr31 into wheat lines and have the potential to provide durable control to stem rust in Australia and elsewhere.

Alleles↗
PubMed2004

Resistance genes for rye stem rust (SrR) and barley powdery mildew (Mla) are located in syntenic regions on short arm of chromosome.

Genetic stocks were developed for the localization and eventual cloning of the stem rust resistance gene SrR that occurs in wheat lines carrying the 1RS translocation from Secale cereale 'Imperial' rye. We have used a mutation-based approach for molecular analysis of the SrR region in rye. Forty-one independent mutants resulting in loss of SrR resistance were isolated: many of these were deletions of various sizes that were used to locate SrR with respect to chromosome group 1S markers. The analysis of the mutants showed that markers about 1 Mb apart flanking the barley Mla locus also flank SrR. Additionally, three of the approximately 20 closely related sequences of Mla in rye are deleted in each of six interstitial deletion mutants of SrR. The results indicate that the SrR region in rye is syntenic to the Mla region in barley or that SrR is possibly orthologous to the Mla locus.

Binding Sites↗
PubMed2001

Contrasting modes of evolution acting on the complex N locus for rust resistance in flax.

Three rust resistance specificities, N, N1 and N2, map to the complex N locus of flax. We used a degenerate PCR approach, with primers directed to the nucleotide binding site (NBS) domain characteristic of many plant resistance genes, to isolate resistance gene analogs (RGAs) from flax. One RGA clone detected RFLPs co-segregating with alleles of the N locus. With this probe we isolated four related genes that occur within a 30kbp region and encode proteins with NBS and leucine-rich repeat (LRR) domains and N-terminal Toll/Interleukin-1 Receptor homology (TIR) domains. One of these four genes was identified as the N resistance gene by sequence analysis of three mutant alleles and by transgenic expression. We isolated homologous genes from two flax lines containing the N1 or N2 specificities and from flax lines carrying no N locus resistance specificities. Analysis of shared polymorphisms among this set of 18 N locus sequences revealed three groups of genes with independent lineages. Sequence exchanges have only occurred between genes within each group, but not between groups. Two of the groups contain only one sequence from each haplotype and probably represent orthologous genes. However, the third group contains two genes from each haplotype. We suggest that the re-assortment of variation by recombination/gene conversion at this locus is limited by the degree of sequence identity between genes.

Alleles↗
PubMed2001

Six amino acid changes confined to the leucine-rich repeat beta-strand/beta-turn motif determine the difference between the P and P2 rust resistance specificities in flax.

At least six rust resistance specificities (P and P1 to P5) map to the complex P locus in flax. The P2 resistance gene was identified by transposon tagging and transgenic expression. P2 is a member of a small multigene family and encodes a protein with nucleotide binding site (NBS) and leucine-rich repeat (LRR) domains and an N-terminal Toll/interleukin-1 receptor (TIR) homology domain, as well as a C-terminal non-LRR (CNL) domain of approximately 150 amino acids. A related CNL domain was detected in almost half of the predicted Arabidopsis TIR-NBS-LRR sequences, including the RPS4 and RPP1 resistance proteins, and in the tobacco N protein, but not in the flax L and M proteins. Presence or absence of this domain defines two subclasses of TIR-NBS-LRR resistance genes. Truncations of the P2 CNL domain cause loss of function, and evidence for diversifying selection was detected in this domain, suggesting a possible role in specificity determination. A spontaneous rust-susceptible mutant of P2 contained a G-->E amino acid substitution in the GLPL motif, which is conserved in the NBS domains of plant resistance proteins and the animal cell death control proteins APAF-1 and CED4, providing direct evidence for the importance of this motif in resistance gene function. A P2 homologous gene isolated from a flax line expressing the P resistance specificity encodes a protein with only 10 amino acid differences from the P2 protein. Chimeric gene constructs indicate that just six of these amino acid changes, all located within the predicted beta-strand/beta-turn motif of four LRR units, are sufficient to alter P2 to the P specificity.

Alleles↗
PubMed2000

Regions outside of the leucine-rich repeats of flax rust resistance proteins play a role in specificity determination.

Multiple alleles controlling different gene-for-gene flax rust resistance specificities occur at the L locus of flax. At least three distinct regions can be recognized in the predicted protein products: the Toll/interleukin-1 receptor homology (TIR) region, a nucleotide binding site (NBS) region, and a leucine-rich repeat (LRR) region. Replacement of the TIR-encoding region of the L6 allele with the corresponding regions of L2 or LH by recombination changed the specificity of the allele from L6 to L7. Replacement of the TIR and most of the NBS-encoding region of L10 with the equivalent region of L2 or L9 generated recombinant alleles having a novel specificity. However, replacement of the L10 TIR-encoding region with the TIR-encoding region of L2 gave rise to an allele with no detectable specificity. These data indicate that non-LRR regions can determine specificity differences between allelic gene products and that functional specificity involves interactions between coadapted polymorphic regions in the protein products of the alleles. Evidence for the action of diversifying selection on the TIR region is observed.

Adaptation, Physiological↗
PubMed1999

Analysis of alternative transcripts of the flax L6 rust resistance gene.

The L6 rust resistance gene from flax generates at least four transcript classes by alternative splicing of the third intron. The most abundant transcript class encodes a resistance protein containing domains that include nucleotide binding site motifs and a leucine-rich repeat region (NBS-LRR). The remaining three transcript classes encode truncated products which lack most of the C-terminal part of the protein containing the leucine-rich region (LRR). The four transcript classes occur in all plant organs examined and no induction of L6 expression was observed following infection of resistant plants with an avirulent rust strain expressing the corresponding A-L6 avirulence gene. Flax plants transgenic for an intronless L6 gene, incapable of encoding truncated resistance proteins by alternative splicing, expressed L6 resistance indistinguishable from that of the wild-type gene. Therefore, a definitive role for alternative transcripts and their predicted truncated products could not be assigned in the flax/flax rust system.

Alternative Splicing↗
PubMed1999

Identification of regions in alleles of the flax rust resistance gene L that determine differences in gene-for-gene specificity.

Thirteen alleles (L, L1 to L11, and LH) from the flax L locus, which encode Toll/interleukin-1 receptor homology-nucleotide binding site-leucine-rich repeat (TIR-NBS-LRR) rust resistance proteins, were sequenced and compared to provide insight into their evolution and into the determinants of gene-for-gene resistance specificity. The predicted L6 and L11 proteins differ solely in the LRR region, whereas L6 and L7 differ solely in the TIR region. Thus, specificity differences between alleles can be determined by both the LRR and TIR regions. Functional analysis in transgenic plants of recombinant alleles constructed in vitro provided further information: L10-L2 and L6-L2 recombinants, encoding the LRR of L2, conferred L2 resistance specificity, and an L2-L10 recombinant, encoding the LRR of L10, conferred a novel specificity. The sequence comparisons also indicate that the evolution of L alleles has probably involved reassortment of variation, resulting from accumulated point mutations, by intragenic recombination. In addition, large deletion events have occurred in the LRR-encoding regions of L1 and L8, and duplication events have occurred in the LRR-encoding region of L2.

Alleles↗
PubMed1998

A flax transposon identified in two spontaneous mutant alleles of the L6 rust resistance gene.

Two spontaneous mutant alleles of the L6 flax rust resistance gene, 16-X3A and 16-X117, contain the same transposable element designated dLute (defective Linum usitatissimum transposable element). The element is 314 bp long, 70% AT-rich and, because it contains no extended open reading frame, is probably non-autonomous. It has 14 bp imperfect terminal inverted repeats related to those in the Ac family of plant transposons and, like Ac, causes 8 bp target site duplications upon insertion. Multiple copies of dLute-related sequences exist in the flax genome. Rust resistant revertants were recovered amongst the progeny of both mutants and reversion was associated with excision of dLute. Excision either restored the wild-type L6 sequence or was imprecise, leaving sequence alterations ('footprints') resulting in one to three amino acid alterations in the L6 protein. No phenotypic differences were discerned between plants containing the standard and revertant L6 alleles.

Alleles↗
PubMed1997

Inactivation of the flax rust resistance gene M associated with loss of a repeated unit within the leucine-rich repeat coding region.

The M rust resistance gene from flax was cloned after two separate approaches, an analysis of spontaneous M mutants with an L6 gene-derived DNA probe and tagging with the maize transposon Activator, independently identified the same gene. The gene encodes a protein of the nucleotide binding site leucine-rich repeat class and is related (86% nucleotide identity) to the unlinked L6 rust resistance gene. In contrast to the L locus, which contains a single gene with multiple alleles, approximately 15 related genes occur at the complex M locus, with only one encoding the M resistance specificity. The M protein contains two direct repeats of 147 and 149 amino acids in the C-terminal part of the leucine-rich region. Three mutant alleles of M encoding a product containing a single repeat unit of 154 amino acids were isolated. The mutant DNA sequences probably occurred by unequal intragenic exchange in the coding region of the repeats. The recombinant alleles lost M resistance and gained no detectable new resistance specificity.

Amino Acid Sequence↗
PubMed1995

Contrasting complexity of two rust resistance loci in flax.

DNA probes from the L6 rust resistance gene of flax (Linum usitatissimum) hybridize to resistance genes at the unlinked M locus, indicating sequence similarities between genes at the two loci. Genetic and molecular data indicate that the L locus is simple and contains a single gene with 13 alleles and that the M locus is complex and contains a tandem array of genes of similar sequence. Thus the evolution of these two related loci has been different. The consequence of the contrasting structures of the L and M loci on the evolution of different rust resistance specificities can now be investigated at the molecular level

Journal Article↗
PubMed1995

The L6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance gene RPS2 and the tobacco viral resistance gene N.

The L6 rust resistance gene from flax was cloned after tagging with the maize transposable element Activator. The gene is predicted to encode two products of 1294 and 705 amino acids that result from alternatively spliced transcripts. The longer product is similar to the products of two other plant disease resistance genes, the tobacco mosaic virus resistance gene N of tobacco and the bacterial resistance gene RPS2 of Arabidopsis. The similarity involves the presence of a nucleotide (ATP/GTP) binding site and several other amino acid motifs of unknown function in the N-terminal half of the polypeptides and a leucine-rich region in the C-terminal half. The truncated product of L6, which lacks most of the leucine-rich C-terminal region, is similar to the truncated product that is predicted from an alternative transcript of the N gene. The L6, N, and RPS2 genes, which control resistance to three widely different pathogen types, are the foundation of a class of plant disease resistance genes that can be referred to as nucleotide binding site/leucine-rich repeat resistance genes.

Alternative Splicing↗
PubMed1994

Heteroduplex molecules formed between allelic sequences cause nonparental RAPD bands.

Primers (10-mers) of random sequence were used to amplify RAPD bands from genomic DNA of an F1 strain of flax rust (Melampsora lini) and its two parent strains. One primer out of 160 tested was unusual in that it amplified a product from F1 DNA that was not amplified from either parental DNAs. The same primer also generated two RAPD bands that segregated as codominant alleles amongst F2 progeny. The nonparental band was only generated from DNAs of F2 individuals that were heterozygous for these two allelic sequences. Sequence analysis of the two RAPD alleles demonstrated greater than 99% sequence identity, although the larger allele possessed an additional 38bp relative to the smaller. Mixing of the two allelic sequences followed by denaturation and annealing in the absence of polymerase activity resulted in the formation of the nonparental band. Thus the nonparental band present in some RAPD reactions consisted of a heteroduplex molecule formed between two allelic sequences of different size. These data demonstrate that heteroduplex molecules formed between allelic RAPD products are a potential source of artifactual polymorphism that can arise during RAPD analysis.

Alleles↗
PubMed1994

Plant resistance to rusts and mildews: genetic control and possible mechanisms.

Genes in plants that confer race-specific resistance to rusts and mildews are widely exploited in agriculture and can prevent huge losses at little cost. However, nothing is known of the molecular basis of their action. Genetic studies, together with observations of responses at the ultrastructural level, can provide broad insights into how resistance is achieved, which may help in cloning resistance genes.

Alleles↗
PubMed1993

Behaviour of modified Ac elements in flax callus and regenerated plants.

The Ac element of maize has been modified by deletion of 537 bases (delta NaeAc) from the untranslated leader of the transposase gene. In a second modification the cauliflower mosaic virus 35S promoter has been inserted into the truncated leader of delta NaeAc, 21 bases upstream of the natural translation start. The activity of these modified elements has been compared with that of the unmodified element in transgenic flax. Deletion of sequences in the untranslated leader only marginally increased transposition in callus while insertion of the 35S promoter enhanced transposition frequency 7-8-fold. Increased transposition correlated with increased transcription of the transposase gene. The presence of a 35S promoter upstream of the transposase gene, but outside the Ac element, also enhanced both transcription and transposition.

Agrobacterium tumefaciens↗
PubMed1987

Cessation of male rat copulatory behavior using illness as punishment: facilitation with a novel odor.

Two experiments were conducted to examine learned copulatory avoidance in male rats. One group of males was presented with receptive females that had been sprayed with a 2% almond solution, and the other group was presented with nonalmond odorous, receptive females. Following each test, males were made ill with lithium chloride (LiCl) by intragastric intubation or intraperitoneal injection. Results showed that male rats presented with almond-odorous females developed significant avoidance of copulatory behavior. Conditioning in males exposed to receptive females without the almond odor developed little, if any, avoidance. In Experiment 2, it was found that route of LiCl administration was not a factor in the results.

Animals↗
PubMedDate not supplied

Neocortical involvement in the acquisition and retention of learned alcohol aversions in rats.

In one experiment, rats lacking gustatory neocortex (GN) were compared with control (both normal and control lesion) rats in the acquisition of a learned alcohol aversion. In a second experiment, the effect of GN ablations on preoperatively learned alcohol aversions was examined. Results showed that rats lacking GN both learned and retained alcohol aversions in a normal manner. These same GN rats, however, extinguished the alcohol aversions significantly faster than controls. These results are in contrast to previous data which indicated that the integrity of the GN was necessary for normal acquisition and retention of learned taste aversions. It is suggested that the odor qualities present in the alcohol stimulus account for the relatively normal performance of GN rats.

Alcohol Drinking↗