scholarly journals Generation and Screening of a BAC Library from a Diploid Potato Clone to Unravel Durable Late Blight Resistance on Linkage Group IV

2007 ◽  
Vol 2007 ◽  
pp. 1-5 ◽  
Author(s):  
Ingo Hein ◽  
Karen McLean ◽  
Boulos Chalhoub ◽  
Glenn J. Bryan
Keyword(s):  
Linkage Group ◽  
Late Blight ◽  
Gene Cluster ◽  
Genomic Library ◽  
Field Resistance ◽  
Group Iv ◽  
Resistance Quantitative Trait Locus ◽  
Diploid Potato ◽  
Bac Clones ◽  
Potato Clone

We describe the construction and screening of a large insert genomic library from the diploid potato clone HB171(13) that has been shown to express durable quantitative field resistance to Phytophthora infestans, the causal agent of potato late blight disease. Integrated genetic mapping of the field resistance quantitative trait locus with markers developed from populations segregating for Rpi-blb3, Rpi-abpt, R2, and R2-like resistance, all located on linkage group IV, has positioned the field resistance QTL within the proximity of this R gene cluster. The library has been successfully screened with resistance gene analogues (RGA) potentially linked to the R gene cluster. Over 30 positive BAC clones were identified and confirmed by PCR and Southern hybridisations to harbour RGA-like sequences. In addition, BAC end sequencing of positive clones has corroborated two BAC clones with a very high level of nucleotide similarity to the RGA probes utilised.

scholarly journals HcrVf paralogs are present on linkage groups 1 and 6 of Malus

Genome ◽  
2009 ◽  
Vol 52 (2) ◽  
pp. 129-138 ◽  
Author(s):  
Giovanni A.L. Broggini ◽  
Paolo Galli ◽  
Gabriella Parravicini ◽  
Luca Gianfranceschi ◽  
Cesare Gessler ◽  
...  
Keyword(s):  
Linkage Group ◽  
Resistance Gene ◽  
Resistance Genes ◽  
Genomic Library ◽  
Sequence Similarity ◽  
Apple Scab ◽  
Scab Resistance ◽  
Transcriptional Analysis ◽  
Linkage Groups ◽  
Bac Clones

Molecular markers derived from resistance gene analogs of HcrVf2, the first apple resistance gene cloned, may pave the way to the cloning of additional apple scab resistance genes. The Malus ×domestica ‘Florina’ (Vf) bacterial artificial chromosome (BAC) genomic library was screened by hybridization using HcrVf2 as a probe. Positive BAC clones were assembled into contigs and microsatellite markers developed from each contig mapped. Only linkage groups 1 and 6 contained HcrVf2 paralogs. On linkage group 1, five loci in addition to the Vf locus were identified. A single locus was detected on linkage group 6. Representative BAC clones of these loci including the Vf locus were sequenced and the gene structure compiled. A total of 22 sequences, showing high sequence similarity to HcrVf2, were identified. Nine sequences were predicted to encode all seven protein domains described in HcrVf2, while three were truncated. Transcriptional analysis indicated that six genes with a complete HcrVf-like structure were constitutively expressed in young uninfected leaves of ‘Florina’. The map position of each HcrVf analog was compared with the location of the major apple scab resistance genes. None of the major genes conferring scab resistance co-localized with HcrVf paralogs, indicating that they are unlikely to belong to the leucine-rich repeat – transmembrane class, which includes the Vf gene.

scholarly journals A Single Gene Cluster Controls Incompatibility and Partial Resistance to Various Melampsora larici-populina Races in Hybrid Poplars

1998 ◽  
Vol 88 (2) ◽  
pp. 156-163 ◽  
Author(s):  
F. Lefèvre ◽  
M. C. Goué-Mourier ◽  
P. Faivre-Rampant ◽  
M. Villar
Keyword(s):  
Gene Cluster ◽  
Partial Resistance ◽  
Random Amplified Polymorphic Dna ◽  
Single Gene ◽  
Field Resistance ◽  
Clonal Variation ◽  
Multiple Alleles ◽  
Hybrid Poplars ◽  
Genetic Background Effect ◽  
Trait Locus

Complete cosegregation for race-specific incompatibility with three Melampsora larici-populina rust races was observed in five F1 hybrid progenies of Populus, with different patterns among the various progenies. A single gene cluster could explain these segregations: one locus with multiple alleles or two tightly linked loci controlling complete resistance to E1 and E3, and two tightly linked loci for E2. The random amplified polymorphic DNA marker OPM03/04_480 was linked to that cluster in all families (<1 cM). This marker accounted for more than 70% of the genetic variation for field resistance in each family (heritability ≈ 0.40). The same marker accounted for up to 64% of the clonal variation for growth in the nursery under natural inoculum pressure; the weak tolerance to rust of F1 interspecific hybrids was attributed to a genetic background effect. Partial resistance was split into epidemiological components (heritability ranged from 0.35 to 0.87). Genotypic correlations among resistance traits for the different races were high (0.73 to 0.90). However, correlations among different resistance components for a single race were not all significant. A major quantitative trait locus for all components of partial resistance to E2 was associated to the cluster controlling incompatibility to E1 and E3 and marked by OPM03/04_480 (R2from 48 to 68%).

scholarly journals Genomic organisation of the Mal d 1 gene cluster on linkage group 16 in apple

2011 ◽  
Vol 29 (3) ◽  
pp. 759-778 ◽  
Author(s):  
Giulia Pagliarani ◽  
Roberta Paris ◽  
Anna Rosa Iorio ◽  
Stefano Tartarini ◽  
Stefano Del Duca ◽  
...  
Keyword(s):  
Linkage Group ◽  
Gene Cluster ◽  
Mal D 1 ◽  
Group 16 ◽  
Genomic Organisation

scholarly journals The Glucuronic Acid Utilization Gene Cluster from Bacillus stearothermophilus T-6

1999 ◽  
Vol 181 (12) ◽  
pp. 3695-3704 ◽  
Author(s):  
Smadar Shulami ◽  
Orit Gat ◽  
Abraham L. Sonenshein ◽  
Yuval Shoham
Keyword(s):  
Gene Cluster ◽  
Transport System ◽  
Bacillus Stearothermophilus ◽  
Glucuronic Acid ◽  
Genomic Library ◽  
Regulatory Gene ◽  
Sugar Transport ◽  
Transcriptional Unit ◽  
Transport Systems ◽  
Potential Operator

ABSTRACT A λ-EMBL3 genomic library of Bacillus stearothermophilus T-6 was screened for hemicellulolytic activities, and five independent clones exhibiting β-xylosidase activity were isolated. The clones overlap each other and together represent a 23.5-kb chromosomal segment. The segment contains a cluster of xylan utilization genes, which are organized in at least three transcriptional units. These include the gene for the extracellular xylanase, xylanase T-6; part of an operon coding for an intracellular xylanase and a β-xylosidase; and a putative 15.5-kb-long transcriptional unit, consisting of 12 genes involved in the utilization of α-d-glucuronic acid (GlcUA). The first four genes in the potential GlcUA operon (orf1, -2, -3, and -4) code for a putative sugar transport system with characteristic components of the binding-protein-dependent transport systems. The most likely natural substrate for this transport system is aldotetraouronic acid [2-O-α-(4-O-methyl-α-d-glucuronosyl)-xylotriose] (MeGlcUAXyl3). The following two genes code for an intracellular α-glucuronidase (aguA) and a β-xylosidase (xynB). Five more genes (kdgK,kdgA, uxaC, uxuA, anduxuB) encode proteins that are homologous to enzymes involved in galacturonate and glucuronate catabolism. The gene cluster also includes a potential regulatory gene, uxuR, the product of which resembles repressors of the GntR family. The apparent transcriptional start point of the cluster was determined by primer extension analysis and is located 349 bp from the initial ATG codon. The potential operator site is a perfect 12-bp inverted repeat located downstream from the promoter between nucleotides +170 and +181. Gel retardation assays indicated that UxuR binds specifically to this sequence and that this binding is efficiently prevented in vitro by MeGlcUAXyl3, the most likely molecular inducer.

scholarly journals Cloning, Sequencing, and Characterization of a Gene Cluster Involved in EDTA Degradation from the Bacterium BNC1

2001 ◽  
Vol 67 (2) ◽  
pp. 688-695 ◽  
Author(s):  
Jan Bohuslavek ◽  
Jason W. Payne ◽  
Yong Liu ◽  
Harvey Bolton ◽  
Luying Xun
Keyword(s):  
Gene Cluster ◽  
Genomic Library ◽  
Regulatory Protein ◽  
Chelating Agent ◽  
Environmental Pollutant ◽  
Flavin Mononucleotide ◽  
Bacterial Strains ◽  
System A ◽  
Monooxygenase Gene ◽  
Genes Encoding

ABSTRACT EDTA is a chelating agent, widely used in many industries. Because of its ability to mobilize heavy metals and radionuclides, it can be an environmental pollutant. The EDTA monooxygenases that initiate EDTA degradation have been purified and characterized in bacterial strains BNC1 and DSM 9103. However, the genes encoding the enzymes have not been reported. The EDTA monooxygenase gene was cloned by probing a genomic library of strain BNC1 with a probe generated from the N-terminal amino acid sequence of the monooxygenase. Sequencing of the cloned DNA fragment revealed a gene cluster containing eight genes. Two of the genes, emoA and emoB, were expressed inEscherichia coli, and the gene products, EmoA and EmoB, were purified and characterized. Both experimental data and sequence analysis showed that EmoA is a reduced flavin mononucleotide-utilizing monooxygenase and that EmoB is an NADH:flavin mononucleotide oxidoreductase. The two-enzyme system oxidized EDTA to ethylenediaminediacetate (EDDA) and nitrilotriacetate (NTA) to iminodiacetate (IDA) with the production of glyoxylate. TheemoA and emoB genes were cotranscribed when BNC1 cells were grown on EDTA. Other genes in the cluster encoded a hypothetical transport system, a putative regulatory protein, and IDA oxidase that oxidizes IDA and EDDA. We concluded that this gene cluster is responsible for the initial steps of EDTA and NTA degradation.

Analysis of lethal mutations induced in a mutator strain that activates transposable elements in Caenorhabditis elegans

Genome ◽  
1990 ◽  
Vol 33 (1) ◽  
pp. 109-114 ◽  
Author(s):  
Denise V. Clark ◽  
Robert C. Johnsen ◽  
Kim S. McKim ◽  
David L. Baillie
Keyword(s):  
Caenorhabditis Elegans ◽  
Linkage Group ◽  
Gamma Ray ◽  
Hot Spot ◽  
Single Gene ◽  
Gene Mutations ◽  
Group Iv ◽  
Group V ◽  
Mutator Strain ◽  
Lethal Mutations

A screen was conducted for lethal mutations in the nematode Caenorhabditis elegans in a strain containing the mutator mut-4(st700)I to examine the nature of mutator-induced lethal mutations within two large chromosomal regions comprising a total of 49 map units (linkage group IV (right) and linkage group V (left)). The genetic analysis of 28 lethal mutations has revealed that the mutator locus mut-4(st700)I causes both putative single-gene mutations and deficiencies. We have identified lethal mutations in three different genes, in addition to seven deficiencies. There is a mutational hot spot on linkage group V (left) around the lin-40 locus. Six mutations appear to be alleles of lin-40. In addition, 5 of 7 deficiencies have breakpoints at or very near lin-40. All seven deficiencies delete the left-most known gene on linkage group V (left) and thus appear to delete the tip of the chromosome. This is in contrast to gamma ray and formaldehyde induced deficiencies, which infrequently delete the closest known gene to the tip of a chromosome.Key words: Caenorhabditis elegans, mutator, deficiencies, lethal mutations.

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