Inheritance, mode of inheritance, and candidate genes for primary hyperparathyroidism in Keeshonden

BACKGROUND: Primary hyperparathyroidism (PHPT) is caused by inappropriate secretion of parathyroid hormone (PTH) by autonomously functioning neoplastic or hyperplastic parathyroid "chief" cells. Keeshonden are thought to be over-represented in studies on canine PHPT, but no proof of heritability or mode of inheritance has been published. The canine disease clinically resembles human familial isolated hyperparathyroidism (FIHP). HYPOTHESIS: Primary hyperparathyroidism in Keeshonden is genetically transmitted and is caused by a mutation in 1 of 4 genes implicated in human FIHP: MEN1, CASR, HRPT2, or RET. ANIMALS: Pedigrees consisting of 1647 Keeshonden were created including 219 Keeshonden with known PHPT phenotypes (69 positive). DNA samples were obtained from 176 of the 219 Keeshonden (34 positive). METHODS: Heritability and mode of inheritance were determined by segregation analysis. Canine homologs to the human genes were identified. Exons and surrounding intron regions were sequenced and scanned for sense-altering polymorphisms or polymorphisms that segregated with the disease. Messenger RNA from a parathyroid tumor of an affected Keeshond was analyzed for polymorphisms and splice alterations. RESULTS: PHPT follows an autosomal dominant mode of inheritance in Keeshonden with possible age-dependent penetrance. No polymorphisms identified in the genes analyzed were associated with a change in predicted protein or in hypothesized splice sites. CONCLUSIONS AND CLINICAL IMPORTANCE: PHPT is an autosomal dominant, genetically transmitted disease in Keeshonden. Once the mutation locus is identified, genetic testing should quickly decrease the incidence of PHPT in this breed. It is unlikely that mutations in MEN1, CASR, HRPT2, or RET cause PHPT in Keeshonden.     

Characterization of Fusarium population associated with wilt of jojoba in Israel

Fusarium wilt, caused by Fusarium oxysporum, is a major disease of jojoba, causing serious economic losses. This study was aimed at characterizing the Fusarium populations associated with jojoba in Israel. Fifty Fusarium isolates used in this study included 23 isolates from the 1990s (“past”) and 27 recently isolated (“recent”). All the isolates were characterized by arbitrarily primed (ap)-PCR and 16 representatives were additionally delineated using multilocus (tef1, rpb1, rpb2) phylogeny and evaluated for their pathogenic potential. Consequently, 88% of the isolates were identified and characterized to the F. oxysporum species complex. The remaining 12% grouped within the F. fujikuroi, F. solani, and F. redolens species complexes. Variations in the infection rate (16.7%–100%), disease symptoms (0.08–1.25, on a scale of 0–3), and fungal colonization index (0.67–2.17, on a scale of 0–4) were observed within the tested isolates, with no significant differences between the past and recent isolates. The representative isolates were assigned to 11 groups based on ap-PCR. Pathogenicity tests showed that isolates from Groups II, IV, and V were the most aggressive, whereas isolates from Groups III, VIII, and IX were the least aggressive. Among the tested isolates, F. oxysporum sensu lato was the most aggressive, followed by F. proliferatum, while F. nygamai was the least aggressive. This study demonstrates the complexity and genetic diversity of Fusarium wilt on jojoba in Israel, indicating possible multiple introductions of infected germplasm into the country.     

Characterization of Pectobacterium brasiliense strains from potato and vegetables in Israel

Pectobacterium brasiliense (Pbr) infects a wide range of crops worldwide, causing potato blackleg and soft rot and vegetable soft rots. This study aimed to characterize the genetic diversity and virulence variability among 68 Pbr strains isolated from either symptomless potato progeny tubers, diseased potato plants, ware potatoes wash water, or vegetables grown in Israel, as well as strains isolated from symptomless seed tubers grown in Europe, or diseased potato plants grown in France. The collection was typed using PCR and TaqMan real-time PCR analyses, dnaX sequence analysis, pulsed-field gel electrophoresis (PFGE), and pectolytic activity. dnaX phylogeny grouped almost all strains in a common genetic clade related to Pbr, which was distinct from the other Pectobacterium species. PFGE analysis identified two main clusters, including one major group of 47 strains with 95%–100% similarity. Maceration assays on two potato cultivars showed significant differences between strains but with no correlations with the source of the strains nor the status of the host (with/without symptoms). Molecular (dnaX sequences and PFGE profiles) and phenotypic analyses (tuber maceration tests) showed that the tested Pbr strains are not a homogeneous group. Analysis of the tested Pbr strains isolated from potato and vegetables grown in fields with a history of potato cultivation suggests that seed tubers imported from Europe may be the main source for Pbr in Israel. To the best of our knowledge, this is the first study that describes biodiversity and population structure of P. brasiliense isolated from potato and vegetables under hot climate conditions.     

Detection of Dickeya spp. latent infection in potato seed tubers using PCR or ELISA and correlation with disease incidence in commercial field crops under hot‐climate conditions

Over a 5‐year period (2006–2010), 277 certified, visually healthy potato seed lots, imported from Europe to Israel for commercial use, were tested for Dickeya spp. latent infection by PCR analysis (277 seed lots) and ELISA (154 seed lots). Seeds from these lots were grown in commercial potato fields which were inspected twice a season by Plant Protection and Inspection Services (PPIS). Stem samples were tested for the presence of Dickeya spp. by PCR analysis. PCR and ELISA results from seed lot testing correlated with disease expression in 74 and 83·8% of the cases, respectively. Positive laboratory results with no disease symptoms in the field (‘+lab/?field’ results) comprised 24·7 and 9·7% of the PCR and ELISA analyses, respectively, whereas negative laboratory results with disease symptoms in the field results (‘?lab/+field’) were obtained in 1·3 and 6·5%, of cases respectively. Maximum disease incidence, as well as the number of cultivars expressing disease symptoms, increased over the years of this study, indicating an increase in the prevalence of the disease. Severe disease incidence was observed on cvs Dita, Rodeo, Desiree, Mondial, Tomensa and Jelly. Of the 55 imported seed lots from which disease was recorded in the field, 49 originated from the Netherlands, four from Germany and two from France. None originated from Scotland.     

A 9-year monitoring study of diseases on potato seed tubers imported to Israel

Potato seed tubers are imported annually to Israel from northern Europe. Although the seed is registered as certified, a survey carried out over a 9-year period indicated that most lots were affected by latent or active bacterial and fungal infections. Latent infection byErwinia carotovora subsp.atroseptica, the causal agent of blackleg, at a level of 10³ cells/g peel, was present in 30% of the lots in most years. Black scurf caused byRhizoctonia solani was present in 20–70% of the imported lots, with a moderate to high level of infection in all years except 1985. In contrast, although many lots were affected by powdery scab, common scab, and Fusarium dry rot in most years, disease incidence within lots was generally low. The gangrene pathogen (Phoma exigua) was rarely detected. The survey findings are of marked importance, due to the extensive use of soil fumigation in Israeli agriculture.     

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase

A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction. The enzyme, isolated from Thermus aquaticus, greatly simplifies the procedure and, by enabling the amplification reaction to be performed at higher temperatures, significantly improves the specificity, yield, sensitivity, and length of products that can be amplified. Single-copy genomic sequences were amplified by a factor of more than 10 million with very high specificity, and DNA segments up to 2000 base pairs were readily amplified. In addition, the method was used to amplify and detect a target DNA molecule present only once in a sample of 10(5) cells.     

Control of potato powdery scab caused by Spongospora subterranea by foliage cover and soil application of chemicals under field conditions with naturally infested soil

Powdery scab (PS), caused by Spongospora subterranea, reduces the quality and marketability of potatoes worldwide. Disease symptoms include lesions on the tuber surface and root galling, which may lead to yield losses. In the current study we report a sustainable approach to reduce PS by manipulating soil temperature during tuber initiation. Plant cover with nonwoven fabric significantly reduced PS on tubers by 54%–69% in 2017 and 84%–93% in 2019, compared to the control, and root galling by 96% in 2019, due to an increased average minimum and maximum soil temperature of 1.8 and 4.2 °C in respective years. Additional preplanting soil treatments were also evaluated in naturally infested soil. In 2017, disease incidence and severity were significantly reduced using 2.5 or 5 L a.i./ha fluazinam in broadcast application or in-furrow, and by 0.375 or 0.75 L a.i./ha flusulphamide applied in-furrow. In 2019, disease incidence and severity were significantly reduced by the broadcast application of fluazinam, and 75 kg/ha calcium cyanamide, but the latter had a negative impact on yield. Soil fumigation with metam sodium resulted in a 98% reduction in PS. Root galling was significantly reduced by calcium cyanamide, metam sodium, and fluazinam in the 2019 trial only. Foliar application of resistance-inducing phosphonates combined with fluazinam application had no additive effect on PS incidence and severity. Integrated approaches such as tolerant cultivars, soil testing, preplanting fungicide application, and sustainable means of control such as foliage cover for a short period may be implemented in order to control the disease and minimize damage.     

Survey of Bacterial and Fungal Seedborne Diseases in Imported and Domestic Potato Seed Tubers

Potato seed tubers are imported to Israel from northern Europe and planted in spring; tubers harvested early from the spring crop are used as seed for the autumn crop. Although only seed lots registered as certified are imported, a previous survey (1984–1994) indicated that most imported lots were affected by latent or active infections caused byErwinia carotovora,Streptomyces scabies, Rhizoctonia solani, Fusarium spp. andSpongospora subterranae. The survey was extended until 1998, and included additional pathogens:Ralstonia solanacearum,Helminthosporium solani, Colletotrichum coccodes andVerticillium dahliae. Most of these pathogens were also monitored in domestic seed tubers, and are reported for the first time. Brown rot was not observed in any of the imported lots. Blackleg and soft rot caused byErwinia spp. were detected in most of the imported lots; however, less than 7% of the lots were contaminated at high levels, while approximately 65% were contaminated at moderate levels. Common scab was detected in most of the imported lots; 51% of the imported lots were contaminated at moderate or high levels, whereas only 6.5% of the domestic seed lots were contaminated at these levels. Black scurf was detected in most of the imported lots; on average, 47.3%, 44.2% and 1.4% of the lots were contaminated at low, moderate and high levels, respectively, and only 7.1% were disease-free. In contrast, most of the domestic lots were either disease-free (45.4%) or had a low disease incidence (37.3%). Only 16.7% of the lots were moderately infected and 0.2% were highly contaminated. Silver scurf was observed in most of the imported lots during all years of the survey, with no differences among the producing countries; on average, 22.7%, 66.1% and 7.5% of the lots were contaminated at low, moderate and high levels, respectively, and only 3.7% were disease-free. Most of the domestic lots (76%) were disease-free and only 6.6% were infected at moderate or high levels. Black dot was observed in a considerable portion of the shipments from Holland during all years of the survey, particularly in 1998, when 34% of the lots were infected. The shipments from France and Germany were infected at low levels, except in 1998, when 19% and 11% of the lots, respectively, arrived infected. In shipments from Scotland and Ireland low incidences of the disease were observed in 1994 and 1995. In the domestic lots, black dot incidence was low (<2.4%) except in 1996, when 11% of the lots were infected.V. dahliae was monitored only in domestic seed tubers. The incidence of disease-free lots was 56–64%, whereas in 20–30% of the lots the level of infection was <5%, and in 6–16% of the lots the level was >5%. The survey findings demonstrate transmission of seedborne pathogens; most of these pathogens can become established in the soil and eventually cause severe outbreaks of disease in potatoes grown in Israel. http://www.phytoparasitica.org posting May 16, 1999.     

Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia

Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia. The first involves the primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase (220,000 times) of target DNA copies. In the second technique, the presence of the beta A and beta S alleles is determined by restriction endonuclease digestion of an end-labeled oligonucleotide probe hybridized in solution to the amplified beta-globin sequences. The beta-globin genotype can be determined in less than 1 day on samples containing significantly less than 1 microgram of genomic DNA.     

Using multi-scale modelling to predict habitat suitability for species of conservation concern: The grey long-eared bat as a case study

Although spatial scale is important for understanding ecological processes and guiding conservation planning, studies combining a range of scales are rare. Habitat suitability modelling has been used traditionally to study broad-scale patterns of species distribution but can also be applied to address conservation needs at finer scales. We studied the ability of presence-only species distribution modelling to predict patterns of habitat selection at broad and fine spatial scales for one of the rarest mammals in the UK, the grey long-eared bat (Plecotus austriacus). Models were constructed with Maxent using broad-scale distribution data from across the UK (excluding Northern Ireland) and fine-scale radio-tracking data from bats at one colony. Fine-scale model predictions were evaluated with radio-tracking locations from bats from a distant colony, and compared with results of traditional radio-tracking data analysis methods (compositional analysis of habitat selection). Broad-scale models indicated that winter temperature, summer precipitation and land cover were the most important variables limiting the distribution of the grey long-eared bat in the UK. Fine-scale models predicted that proximity to unimproved grasslands and distance to suburban areas determine foraging habitat suitability around maternity colonies, while compositional analysis also identified unimproved grasslands as the most preferred foraging habitat type. This strong association with unimproved lowland grasslands highlights the potential importance of changes in agricultural practices in the past century for wildlife conservation. Hence, multi-scale models offer an important tool for identifying conservation requirements at the fine landscape level that can guide national-level conservation management practices.