Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Tree and forest encroachment into fescue grasslands on the Cypress Hills plateau,southeast Alberta,Canada

Tree encroachment into rough fescue (Festuca campestris) grassland has been identified as an ecological concern on the Cypress Hills plateau in southeastern Alberta, Canada. A combination of field sampling (109 transects), a dendrochronological assessment (1361 trees), and a time series analysis of remotely sensed images from five different time periods (1950–2002) were used to determine the extent and rate of tree encroachment and forest development. Tree cover increased by 768 ha (～51%) between 1950 and 2007, representing 10% of the study area, from 1502 ha of pre-1950 forest. Post-1950 tree invasion also created an ingress zone of 750 ha (～10% of study area) based on field transects. Forest cover increased at a consistent rate of 14.3 ha/year. Lodgepole pine (Pinus contorta var. latifolia) was the most common tree species associated with encroachment. Invasion based on the number of established trees occurred at an exponential rate of 3.1%/year after 1890, with density increased by filling spaces adjacent to and between trees within the grassland vegetation. The rate of tree establishment increased to 4.4%/year after 1980, suggesting a change occurred in environmental conditions. Annual atmospheric temperatures increased 0.55 °C from 1929 to 2005 (P < 0.001), with winter (December–February) and spring (March–May) temperatures accounting for most of the increase (P < 0.001), whereas summer (June–August) temperatures slightly decreased (P < 0.050, 0.34 °C) and precipitation increased (P < 0.005, 30 mm). Cattle and elk (Cervus elaphus) grazing was not considered a primary factor for explaining tree encroachment. Based on multidimensional scaling, lodgepole pine establishment was associated with warmer spring temperatures and greater fall (October–November) precipitation. A landscape devoid of wildfires, combined with greater moisture availability, and a longer frost-free season is likely conducive to the sustained establishment of coniferous trees and forest development within the Cypress Hills fescue grassland ecosystem. Long-term conservation of the fescue grasslands could be possible by reintroducing fire.     

In vitro insecticidal effects of fipronil and beta-cyfluthrin on larvae of the blowfly Lucilia sericata

The insecticidal effects of the phenylpyrazole, fipronil, and a pyrethroid, beta-cyfluthrin, on larvae of the blowfly Lucilia sericata were determined in laboratory assays. When first stage larvae of L. sericata were reared on homogenized pig liver which had been treated with known amounts of test compounds, both fipronil and beta-cyfluthrin induced significant levels of mortality compared to acetone and water controls. However, fipronil was approximately 10 times more toxic than beta-cyfluthrin to L. sericata larvae following ingestion. Beta-cyfluthrin had little effect on mortality until concentrations of approximately 0.5 ppm were reached. In contrast, fipronil effected L. sericata mortality at a concentration of 0.05 ppm and 100% mortality was reached by 0.5 ppm. The lethal concentration (LC50) value for beta-cyfluthrin was 1.56 ppm as compared to 0.14 ppm for fipronil. Following contact of first and third stage larvae with cloth impregnated with known amounts of test compound, the mortality profiles of fipronil and beta-cyfluthrin were similar. At short contact times, the LC50 values for fipronil were lower than those for beta-cyfluthrin. However, at the highest contact time evaluated for the first stage larvae, 300 s, there was a reversal in this trend. The results suggest that the phenylpyrazole fipronil may represent a new potential insecticide for development against blowfly strike of sheep.     

Dysgenesis of the tubal system in an infertile mare karyotype 63,X:64,XY:65,XXY

A Thoroughbred mare, barren during three years at stud, unresponsive to therapy, and not palpably abnormal anatomically was found to have hypoplasia of the uterine tubes after excision at laparotomy. The cause was an unbalanced genotype. The mosaic karyotype, determined on a blood sample, was indispensible in diagnosis and the case extends the potential for cytogenetic prognostic procedures.