Host Specialization not Detected Among Isolates of the EC-1 Lineage of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> Attacking Wild and Cultivated Potatoes in Peru

To determine whether populations of Phytophthora infestans attacking wild and cultivated potatoes in the highlands of Peru are specialized on their hosts of origin, we characterized isolates using several neutral markers, metalaxyl resistance and for aggressiveness in a detached leaf assay. One hundred and fifty-three isolates were collected from the northern and central highlands of Peru from different potato cultivars (both modern and native cultivars) and from different species of wild, tuber-bearing potatoes. All the isolates analyzed belonged to one of four clonal lineages that had been described previously in Peru: EC-1, US-1, PE-3 and PE-7. The EC-1 lineage (n = 133) was dominant and present in similar frequencies on wild and cultivated potatoes. PE-3 (n = 14) was found primarily on cultivated potatoes, with only one isolate coming from a wild host. US-1 (n = 2) and PE-7 (n = 4) were rare; all but one (PE-7) occurred on wild potatoes. Isolates from the EC-1 lineage from modern cultivars were compared in three separate detached leaf inoculation assays with EC-1 isolates from the wild potato species S. sogarandinum, S. bill-hookerii or S. huancabambense, respectively. No significant interactions between isolate type (from wild or cultivated potato) and host type (wild or cultivated) were measured for any assay. It appears that the pathogen genotypes in the EC-1 lineage indiscriminately attack both wild and cultivated tuber-bearing solanaceous hosts in Peru, and breeders should be able to select for resistance using the common EC-1 lineage.     

Evidence for morphological, vegetative, genetic, and mating-type diversity in Sclerotinia homoeocarpa

Morphology, vegetative compatibility groups, and molecular characteristics were compared among 47 isolates of the dollar spot pathogen Sclerotinia homoeocarpa. Isolates were collected from cool- and warm-season turfgrasses in Florida and the northern United States. Mycelial pigment accumulation, substratal stromata formation, and symptom development were used to separate the collection into two distinct morphological types: a common-type (C-type) and a Floridian-type (F-type). Phylogenetic relationships estimated from ITS sequences supported the morphological typing. Identification and characterization of the S. homoeocarpa mating-type locus revealed an idiomorphic organization for both C- and F-types with nearly equal frequencies of each mating types present in both groups. These findings suggest heterothallic control of mating and indicate potential for outcrossing in both groups. Dollar spot disease of turfgrass in Florida is caused by two distinct morphological types of S. homoeocarpa which may be cryptic species. These findings could have implications for disease management.     

Seawater tolerance and morphological assessment of yearling sea trout (Salmo trutta L.)

To study seawater tolerance and make a morphological assessment of yearling sea trout, their maturation and smoltification signs were estimated in two different fish rearing systems in Latvia,—in recirculating and flow-through system. For yearling sea trout (Salmo trutta L.), fish hypo-osmoregulatory ability was evaluated using seawater tolerance test, also fish morphological parameters were analyzed from January to May. April and May are months when smoltification occurs for wild sea trout in nature. Sea trout from recirculating system initially showed better growth, higher survival rate, and hypo-osmoregulatory ability, due to the elevated rearing water temperature. However, the situation completely changed in May when natural smoltification peaked and wild sea trout migration to the sea occurred—survival rate for yearling sea trout reared in recirculating system dropped to zero. Nevertheless, survival of fish from flow-through system increased, reaching 33%. Furthermore, in flow-through systems, sea trout that survived had significantly lower condition factor, also silvering level was higher compared those who did not survive. Weight was not a useful factor for determination of sea trout smoltification. The most part of yearling sea trout did not smoltify at the age of 1 year and should be reared for one more year before release in natural watercourses foreseen for migration to the sea.

     

Germline mutations in genes within the MAPK pathway cause cardio-facio-cutaneous syndrome

Cardio-facio-cutaneous (CFC) syndrome is a sporadic developmental disorder involving characteristic craniofacial features, cardiac defects, ectodermal abnormalities, and developmental delay. We demonstrate that heterogeneous de novo missense mutations in three genes within the mitogen-activated protein kinase (MAPK) pathway cause CFC syndrome. The majority of cases (18 out of 23) are caused by mutations in BRAF, a gene frequently mutated in cancer. Of the 11 mutations identified, two result in amino acid substitutions that occur in tumors, but most are unique and suggest previously unknown mechanisms of B-Raf activation. Furthermore, three of five individuals without BRAF mutations had missense mutations in either MEK1 or MEK2, downstream effectors of B-Raf. Our findings highlight the involvement of the MAPK pathway in human development and will provide a molecular diagnosis of CFC syndrome.     

Mycobacterial Ku and ligase proteins constitute a two-component NHEJ repair machine

In mammalian cells, repair of DNA double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ) is critical for genome stability. Although the end-bridging and ligation steps of NHEJ have been reconstituted in vitro, little is known about the end-processing reactions that occur before ligation. Recently, functionally homologous end-bridging and ligation activities have been identified in prokarya. Consistent with its homology to polymerases and nucleases, we demonstrate that DNA ligase D from Mycobacterium tuberculosis (Mt-Lig) possesses a unique variety of nucleotidyl transferase activities, including gap-filling polymerase, terminal transferase, and primase, and is also a 3' to 5' exonuclease. These enzyme activities allow the Mt-Ku and Mt-Lig proteins to join incompatible DSB ends in vitro, as well as to reconstitute NHEJ in vivo in yeast. These results demonstrate that prokaryotic Ku and ligase form a bona fide NHEJ system that encodes all the recognition, processing, and ligation activities required for DSB repair.     

Malignant activation of a K-ras oncogene in lung carcinoma but not in normal tissue of the same patient

A single genetic alteration, a guanine-to-cytosine transversion, is responsible for the acquisition of malignant properties by K-ras genes of two human tumor cell lines established from carcinomas of the bladder (A1698) and lung (A2182). As a consequence, arginine instead of the normal glycine is incorporated into the K-ras-coded p21 proteins at amino acid position 12. This mutation creates a restriction enzyme polymorphism that can be used to screen human cells for transforming K-ras genes. This approach was used to identify the mutational event responsible for the malignant activation of a K-ras oncogene in a squamous cell lung carcinoma of a 66-year-old man; this point mutation was not present in either the normal bronchial or parenchymal tissue or in the blood lymphocytes. Hence, malignant activation of a ras oncogene appears to be specifically associated with the development of a human neoplasm.     

Identification of a DNA nonhomologous end-joining complex in bacteria

In eukaryotic cells, double-strand breaks (DSBs) in DNA are generally repaired by the pathway of homologous recombination or by DNA nonhomologous end joining (NHEJ). Both pathways have been highly conserved throughout eukaryotic evolution, but no equivalent NHEJ system has been identified in prokaryotes. The NHEJ pathway requires a DNA end-binding component called Ku. We have identified bacterial Ku homologs and show that these proteins retain the biochemical characteristics of the eukaryotic Ku heterodimer. Furthermore, we show that bacterial Ku specifically recruits DNA ligase to DNA ends and stimulates DNA ligation. Loss of these proteins leads to hypersensitivity to ionizing radiation in Bacillus subtilis. These data provide evidence that many bacteria possess a DNA DSB repair apparatus that shares many features with the NHEJ system of eukarya and suggest that this DNA repair pathway arose before the prokaryotic and eukaryotic lineages diverged.