Genetic Differentiation of Puccinia triticina Populations in Central Asia and the Caucasus

ABSTRACT Isolates of Puccinia triticina collected from common wheat in the Central Asia countries of Kazakhstan, Uzbekistan, Tajikistan, and Kyrgyzstan and the Caucasus countries of Azerbaijan, Georgia, and Armenia were tested for virulence to 20 isolines of Thatcher wheat with different leaf rust resistance genes and molecular genotype at 23 simple sequence repeat (SSR) loci. After clone correction within each country, 99 isolates were analyzed for measures of population diversity, variation at single SSR loci, and for genetic differentiation of virulence phenotypes and SSR genotypes. Isolates from Central Asia and the Caucasus were also compared with 16 P. triticina isolates collected from common wheat in North America that were representative of the virulence and molecular variation in this region and two isolates collected from durum wheat in France and the United States. Populations from the Caucasus, Uzbekistan, Tajikistan, and Kyrgyzstan were not significantly (P > 0.05) differentiated for SSR variation with F(st) and R(st) statistics. Populations from the Caucasus, Uzbekistan, Tajikistan, and Kyrgyzstan were significantly (P < 0.05) differentiated from the populations in South and North Kazakhstan for SSR variation. All populations from Central Asia and the Caucasus were significantly differentiated from the North American isolates and isolates from durum wheat for SSR variation and virulence phenotypes. There was a correlation between virulence phenotype and SSR genotype among individual isolates and at the population level. Mountain barriers may account for the differentiation of P. triticina geographic populations in Central Asia and the Caucasus.     

Adherence of Actinobacillus pleuropneumoniae serotype 1 to swine buccal epithelial cells involves fibronectin

The swine pathogen Actinobacillus pleuropneumoniae serotype 1 was investigated for its ability to adhere to swine, rat, and human buccal epithelial cells (BEC). The highest number of bacteria adhered was to swine BEC. This binding ability was affected by heating, extreme pH, treatment with sodium dodecyl sulfate, ethylenediamine tetraacetate, or periodate, and proteolysis, suggesting that cell-surface glycoproteins participate in adherence and that adherence is based mostly on ionic interactions. Mannose and swine fibronectin may play a direct role in this interaction. Convalescent-phase serum from naturally infected pigs inhibited the adhesion. There was a correlation between bacterial pathogenicity as well as host specificity and the capacity for adherence to swine BEC. Adhesion to swine BEC provides a convenient method to study in vitro the adherence of A. pleuropneumoniae and other pathogens of the pig respiratory tract.     

Variation of pathotypes and races and their correlations with clonal lineages in Verticillium dahliae

Understanding pathogenic variation in plant pathogen populations is key for the development and use of host resistance for managing verticillium wilt diseases. A highly virulent defoliating (D) pathotype in Verticillium dahliae has previously been shown to occur only in one clonal lineage (lineage 1A). By contrast, no clear association has yet been shown for race 1 with clonal lineages. Race 1 carries the effector gene Ave1 and is avirulent on hosts that carry resistance gene Ve1 or its homologues. The hypothesis tested was that race 1 arose once in a single clonal lineage, which might be expected if V. dahliae acquired Ave1 by horizontal gene transfer from plants, as hypothesized previously. In a diverse sample of 195 V. dahliae isolates from nine clonal lineages, all race 1 isolates were present only in lineage 2A. Conversely, all lineage 2A isolates displayed the race 1 phenotype. Moreover, 900‐bp nucleotide sequences from Ave1 were identical among 27 lineage 2A isolates and identical to sequences from other V. dahliae race 1 isolates in GenBank. The finding of race 1 in a single clonal lineage, with identical Ave1 sequences, is consistent with the hypothesis that race 1 arose once in V. dahliae. Molecular markers and virulence assays also confirmed the well‐established finding that the D pathotype is found only in lineage 1A. Pathogenicity assays indicated that cotton and olive isolates of the D pathotype (lineage 1A) were highly virulent on cotton and olive, but had low virulence on tomato.     

Soybean molasses as an organic carbon source in the farming of Litopenaeus vannamei (Boone, 1931) in a biofloc system

Soybean molasses was evaluated as a partial replacement for sugarcane molasses as a carbon source for biofloc development in the superintensive culture of Pacific white shrimp (Litopenaeus vannamei). A 50‐day study was conducted with juvenile (3.2 g) shrimp stocked in 16 800 L tanks at a stocking density of 250 shrimp m^?3. Control of total ammonia concentration was performed by the addition of combined mixtures of soybean and sugarcane molasses to the culture water. Three different molasses treatments were evaluated using different soybean‐to‐sugarcane molasses ratios: 15–85%, 38–62% and 60–40% respectively. The control group was treated only with sugarcane molasses. Water quality, chlorophyll a concentration, heterotrophic bacterial load, Vibrio spp. concentration and zootechnical indexes were all evaluated. Total ammonia concentration was controlled by heterotrophic and chemotrophic pathways. Biofloc formation, as quantified by measuring the total suspended solids, was not altered. The Vibrio spp. concentration showed a significant reduction in treatments with soybean‐to‐sugarcane molasses ratios of 38–62% and 60–40%. All combined mixtures of soybean and sugarcane molasses could maintain water quality and productivity in the superintensive culture of L. vannamei using the biofloc system. Thus, the potential use of a residue from agroindustry as a carbon source in a biofloc culture is demonstrated.     

In vitro morphogenesis of juvenile Annona cherimola Mill,bud explants

A micropropagation procedure for juvenile cherimoya (Annona clierimola Mill.) is described. Axillary shoot proliferation was obtained after culturing nodal sections with lateral buds in basal medium supplemented with either 0.66 μM BA or 1,36μM zeatin. For root induction shoots were pre-incubated for 7 d in basal medium supplemented with 1 gl^?1 activated charcoal, then cultured for 10 d (7 dark/3 light) on medium with 500 μM IBA, 58.4 mM sucrose, and 200 mg I“1 citric acid. Afterwards, shoots were transferred to the same medium without auxin and with the macroelements at half strength. Using this procedure, 95% of shoots rooted. The survival rate at the end of the acclimatization period was 70%.     

Cadmium Effects in Sunflower: Nutritional Imbalances in Plants and Calluses

The effects of cadmium (Cd) exposure on sunflower (Helianthus annuus L.) nutrient accumulation remain unclear. However, studies concerning crop improvement for Cd tolerance suggest the use of biotechnology techniques such as tissue culture. It is still unknown whether in vitro cells respond to Cd exposure in a way similar to plants. In this paper, the objectives were (1) to characterize the effects of Cd exposure in macronutrient and micronutrient accumulation in different sunflower organs/tissues and (2) to compare the behavior of two culture systems (plants vs. tissue culture) regarding Cd and nutrient accumulation. To achieve these aims, sunflower plants were grown hydroponically in the presence of Cd (at levels of 0, 5, 50, and 500 μ M). For in vitro cultures, seeds were germinated axenically and leaf explants were then grown on Murashige and Skoog medium (MS). One-month-old calluses were grown on MS medium containing 0, 5, 50, and 500 μ M Cd. After 21 d of exposure to 500 μ M, all plants were dead. The contents of macro- and micronutrients and of Cd were determined by ICPS in 18 d-exposed plants and calluses and in calluses exposed for six months to 50 μ M Cd. At day 18, Cd content increased in leaves, roots, and calluses. Cadmium exposure also decreased the contents of magnesium (Mg), calcium (Ca), iron (Fe), and manganese (Mn) in roots and of Mg, Ca, copper (Cu), Fe, and Mn in shoots. Exposed calluses suffered decreases only in Mg, Ca, and Mn contents. The contents of most of these nutrients in six-month-exposed calluses were similar to those of the control calluses, indicating that these long-term exposed in vitro cells developed mechanisms for regulating the effects of Cd on the accumulation of nutrients.     

Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Contrasting phenology and female cone characteristics of the two Macaronesian island endemic cedars (<Emphasis Type="Italic">Juniperus cedrus</Emphasis> and <Emphasis Type="Italic">J. brevifolia</Emphasis>)

Phenology and female cone characteristics of the two endemic cedars (Juniperus cedrus and J. brevifolia) from the Macaronesian islands were studied. Despite their closely taxonomic affinity and their evolution under insular conditions, different trends were recorded. Mature J. cedrus female cones were present throughout the year, while those from J. brevifolia were only present in summer and autumn. J. cedrus female cone size was significantly larger than that of J. brevifolia, a trend consistent with the presence of larger vertebrates (lizards and birds) in the Canary Islands. However, water content was four times higher in J. brevifolia female cones, which can be related with the higher rainfall existing in the Azores. J. cedrus has two or three seeds per cone, whereas J. brevifolia frequently had three. Seeds from J. cedrus were clearly larger and heavier, coinciding with the female cone size trend. However, tetrazolium tests revealed higher viability values in J. brevifolia. The relatively low percentage of filled seeds in J. cedrus could be a consequence of the climatic stress and limits to pollination due to fragmented populations as described for other Juniperus species. In summary, our results reveal that some environmental factors such as the harsh conditions, high population fragmentation and the dependence on large dispersers have compromised the fitness of J. cedrus in the Canary Islands.     

Natural hybridization between wheat (Triticum aestivum L.) and its wild relatives Aegilops geniculata Roth and Aegilops triuncialis L.

BACKGROUND

Cultivated bread wheat (Triticum aestivum L.) spontaneously hybridizes with wild/weedy related Aegilops populations, but little is known about the actual rates at which this hybridization occurs under field conditions. It is very important to provide reliable empirical data on this phenomenon in order to assess the potential crop–wild introgression, especially in the context of conducting risk assessments for the commercialization of genetically modified (GM) wheat, as gene flow from wheat to Aegilops species could transfer into the wild species genes coding for traits such as resistance to herbicides, insects, diseases or environmental stresses.

RESULTS

The spontaneous hybridization rates between wheat and A. geniculata and A. triuncialis, which are very abundant in the Mediterranean area, have been estimated for the first time in the northern part of the Meseta Central, the great central plateau which includes the largest area of wheat cultivation in Spain. Hybridization rates averaged 0.12% and 0.008% for A. geniculata and A. triuncialis, respectively. Hybrids were found in 26% of A. geniculata and 5% of A. triuncialis populations, at rates that can be ≤3.6% for A. geniculata and 0.24% for A. triuncialis.

CONCLUSION

The detection of Aegilops spp.–wheat hybrids in Aegilops populations indicates that gene flow can occur, although wheat is considered a crop with a low-to-medium risk for transgene escape. These data on field hybridization rates are essential for GM wheat risk assessment purposes. © 2023 Society of Chemical Industry.     

Molecular characterization and phylogenetic analysis of Newcastle disease virus isolates from healthy wild birds

Wild waterfowl is considered a natural reservoir of potentially infectious agents and a source of pathogenic viruses like avian paramyxoviruses type 1 (APMV 1). In 1997, commercial poultry in Argentina had reached the status of being free from virulent Newcastle disease virus (NDV) infections. Vaccination and biosecurity measures are actively performed to maintain this preferential sanitary condition. However, the risk of reintroduction of pathogenic viruses is always present. In this context, we conducted a study to describe the status of wild healthy birds in a geographic region relevant for the poultry industry. The presence of anti-NDV antibodies was determined in different species in all areas sampled suggesting previous contact with NDV. Seven ND viruses were isolated and characterized as apathogenic strains by biological and molecular methods. The phylogenetic analysis revealed that the majority of the Argentinian isolates form a subgroup related to viruses of genotype II. The results presented here highlight the importance of maintaining strict biosecurity measures and vaccination programs in poultry industries in order to preserve the virulent NDV-free status for commercial flocks in the country.