Soil characteristics below <Emphasis Type="Italic">Erythrina poeppigiana</Emphasis> in organic and conventional Costa Rican coffee plantations

The impact of Erythrina poeppigiana on soil characteristics, at three different positions relative to the shade tree and from three different soil depths, was evaluated in pairs of comparable Costa Rican coffee farms (organic and conventional) in 2000 and 2004. In the conventional system at 0–5 cm, higher C and N concentrations were found close to the shade tree versus the positions 2 m from the trunk (5.04 vs. 4.18%). This positive effect could influence only 20% of the farm area when high population of E. poeppigiana were used. This finding highlighted the importance of E. poeppigiana in maintaining SOM levels. In contrast, the organic system showed similar C and N concentrations for all positions probably due to an even distribution of pruning residues and to the use of organic amendments. A trend to higher total C and N concentrations for organic farms in comparison to conventional farms was found. No significant temporal changes in soil C or N concentrations were found between 2000 and 2004.     

A low cost field‐survey method for mapping seagrasses and their potential threats: an example from the northern Gulf of Aqaba,Red Sea

1. In the Gulf of Aqaba (GoA), coral reefs are considered the dominating ecosystem, while seagrass meadows, recognized worldwide as important ecosystems, have received little attention. Absence of comprehensive seagrass maps limits awareness, evaluations of associated ecosystem services, and implementation of conservation and management tools. 2. Presented here are the first detailed maps of seagrass meadows along the Israeli coast of the northern GoA. Mapping was performed by snorkelling along transects perpendicular to the shore above meadows growing at 15–25 m. Measurements along these transects included position, meadow depth and visual estimations of seagrass cover. Shallow boundaries of meadows, parallel to shore, were recorded by GPS tracking. Supplementary work included drop‐camera boat surveys to determine the position of the deeper edge of meadows. In addition, GIS layers were created that indicated shoreline infrastructures, near‐shore human activities and potential pollution threats. Ecosystem services of seagrass meadows mapped were valuated using a benefit transfer approach. 3. In total, 9.7 km of the 11 km shoreline were surveyed and 2830 data points collected. Seagrasses were growing along 7.5 km of the shoreline, with shallow (15–25 m) meadows found to cover an area of 707 000 m² and valued at more than US$ 2 000 000 yr^‐1 in associated ecosystem services. Pilot drop‐camera surveys (additional 283 data points) indicated that meadows can extend down to 50 m in some places. Coastal uses and threats varied in character and location. A municipality runoff point and drainage canal located close to the largest meadow were identified as the main threats to local seagrasses. 4. These low‐cost methods enhance our understanding of seagrass distribution in the northern GoA. They demonstrate a GIS‐based tool for assessing how environmental changes might affect the cover and state of seagrasses, improving efforts to conserve seagrass, and have particular relevance to seagrass mapping in developing countries and/or island nations. Copyright © 2016 John Wiley & Sons, Ltd.     

A fragment of the Xanthomonas oryzae pv. oryzicola harpin HpaG Xooc reduces disease and increases yield of rice in extensive grower plantings

Harpins of phytopathogenic bacteria stimulate defense and plant growth in many types of plants, conferring disease resistance and enhanced yield. In a previous study, we characterized nine fragments of the harpin protein HpaG(Xooc) from Xanthomonas oryzae pv. oryzicola for plant defense elicitation and plant growth stimulation activity relative to the intact protein. In plants grown under controlled conditions, the fragment HpaG10-42 was more active in both regards than HpaG(Xooc). Here, we demonstrate that the activity of HpaG10-42 in rice under field conditions significantly exceeds that of HpaG(Xooc), stimulating resistance to three important diseases and increasing grain yield. We carried out tests in 672 experimental plots with nine cultivars of rice planted at three locations. Application protocols were optimized by testing variations in application rate, frequency, and timing with respect to rice growth stage. Of the concentrations (24, 24, 12, and 6 microg/ml), and number and timing of applications (at one to four different stages of growth) tested, HpaG10-42 at 6 microg/ml applied to plants once at nursery seedling stage and three times in the field was most effective. Bacterial blight, rice blast, and sheath blight were reduced 61.6 and 56.4, 93.6 and 76.0, and 93.2 and 55.0% in indica and japonica cultivars, respectively, relative to controls. Grain yields were 22 to 27% greater. These results are similar to results obtained with typical local management practices, including use of chemicals, to decrease disease severities and increase yield in rice. Our results demonstrate that the HpaG10-42 protein fragment can be used effectively to control diseases and increase yield of this staple food crop.     

Identification of specific fragments of HpaG Xooc, a harpin from Xanthomonas oryzae pv. oryzicola, that induce disease resistance and enhance growth in plants

Harpin proteins from gram-negative plant-pathogenic bacteria can stimulate hypersensitive cell death (HCD) and pathogen defense as well as enhance growth in plants. Two of these diverse activities clearly are beneficial and may depend on particular functional regions of the proteins. Identification of beneficial and deleterious regions might facilitate the beneficial use of harpin-related proteins on crops without causing negative effects like cell death. Here, we report the identification and testing of nine functional fragments of HpaG(Xooc), a 137-amino-acid harpin protein from Xanthomonas oryzae pv. oryzicola, the pathogen that causes bacterial leaf streak of rice. Polymerase chain reaction-based mutagenesis generated nine proteinaceous fragments of HpaG(Xooc); these caused different responses following their application to Nicotiana tabacum (tobacco) and Oryza sativa (rice). Fragment HpaG62-137, which spans the indicated amino acid residues of the HpaG, induced more intense HCD; in contrast, HpaG10-42 did not cause evident cell death in tobacco. However, both fragments stimulated stronger defense responses and enhanced more growth in rice than the full-length parent protein, HpaG(Xooc). Of the nine fragments, the parent protein and one deletion mutant of HpaG(Xooc) tested, HpaG10-42, stimulated higher levels of rice growth and resulted in greater levels of resistance to X. oryzae pv. oryzae and Magnaporthe grisea. These pathogens cause bacterial leaf blight and rice blast, respectively, the two most important diseases of rice world-wide. HpaG10-42 was more active than HpaG(Xooc) in inducing expression of several genes that regulate rice defense and growth processes and activating certain signaling pathways, which may explain the greater beneficial effects observed from treatment with that fragment. Overall, our results suggest that HpaG10-42 holds promise for practical agricultural use to induce disease resistance and enhance growth of rice.     

Perspectives on molecular detection methods of lyssaviruses

Lyssaviruses belong to the family Rhabdoviridae within the order Mononegavirales. Their genomes consist of a negative stranded linear RNA. Although dog transmitted rabies causes the majority of the estimated 55 000 worldwide fatalities per year, bat lyssaviruses have also caused human cases. Based on a broad range of different host species, their geographical distribution and the evolutionary age lyssaviruses display a high degree of genetic variability. Thus the development of molecular diagnostics for the reliable detection and identification of members of the genus Lyssavirus (pan-lyssavirus assay) is a challenge, but is valuable for the fight against rabies worldwide. In this study different published PCR-systems or primers for the detection of a wide range of lyssaviruses were tested as real-time RT-PCRs with different one-step and two-step assays. Comparison of different two-step assays by varying the RT-chemistry revealed differences in sensitivity. Nevertheless, most of the tested one-step systems provided an improved performance, including a reduced assay time and a reduced risk of cross-contaminations, when compared to the optimized two-step assay. Finally, we also provide an overview of additional state-of-the-art molecular methods to detect and differentiate lyssavirus species in general.     

Epizootic hemorrhagic disease virus infection of type I interferon receptor deficient mice

Type I interferon receptor deficient (IFNAR(-/-)) mice were infected with an Israeli isolate of serotype 7 of epizootic hemorrhagic disease virus (EHDV; Orbivirus, Reoviridae). Two out of two mice that received 5×10(5) 50% tissue culture infectious doses (TCID(50)) by intraperitoneal injection died or were euthanized in a moribund state on day 5 after infection. One mouse out of three that had been inoculated with 5×10(2) TCID(50) died on day 7 while the remaining mice did not show any clinical signs and survived until the end of the experiment. Spleens of all dead mice were highly positive in an EHDV real-time RT-PCR (quantification cycle values ≤15) and contained ≥10(5.8) TCID(50) of virus per ml of homogenate. The viral RNA content and virus titer in the spleens of the two surviving mice, on the other hand, were over 100-fold lower. Different from data reported for BTV, the outcome of EHDV infection of IFNAR(-/-) mice is dose-dependent and subclinical infections can occur.     

Effect of heat on aspalathin, iso-orientin, and orientin contents and color of fermented rooibos (Aspalathus linearis) iced tea

The phenolic quality of commercial South African fermented rooibos iced teas in terms of aspalathin, iso-orientin, and orientin contents in comparison to a "cup of tea" was shown to be inferior. The role of the different manufacturing stages of powdered extract used in iced tea formulation and, more specifically, the impact of pasteurization and sterilization on the color and phenolic content of the beverage, were assessed as potential causes of its inferior phenolic quality. Aspalathin and its corresponding flavones, iso-orientin and orientin, were found to be present at all stages of the powdered extract production process. Spray-drying did not significantly (P ≥ 0.05) alter the aspalathin, iso-orientin, or orientin content of concentrates. Simulated normal-temperature sterilization (NTS at 121 °C/15 min) and high-temperature sterilization (HTS at 135 °C/4 min), but not necessarily pasteurization (93 °C/30 min), significantly (P < 0.05) reduced the aspalathin, iso-orientin, and orientin contents of different iced tea formulations. Heat-induced losses of iso-orientin and orientin were lower than those for aspalathin. Conversion of aspalathin to the flavones is implicated. The addition of ascorbic acid and/or citric acid to the base iced tea formulation containing only rooibos extract and sugar proved to be beneficial, especially for the retention of aspalathin. Browning, that is, absorbance at 420 nm, was significantly (P < 0.05) increased in the base formulation. In the case of the formulations also containing ascorbic acid and/or citric acid, absorbance remained unchanged or decreased when subjected to NTS and HTS treatments. This was attributed to removal of brown polymers from solution as the pH values of these formulations were lower than that of the base formulation.     

Pea Aphids (Hemiptera: Aphididae) Have Diurnal Rhythms When Raised Independently of a Host Plant

Seasonal timing is assumed to involve the circadian clock, an endogenous mechanism to track time and measure day length. Some debate persists, however, and aphids were among the first organisms for which circadian clock involvement was questioned. Inferences about links to phenology are problematic, as the clock itself is little investigated in aphids. For instance, it is unknown whether aphids possess diurnal rhythms at all. Possibly, the close interaction with host plants prevents independent measurements of rhythmicity. We reared the pea aphid Acyrthosiphon pisum (Harris) on an artificial diet, and recorded survival, moulting, and honeydew excretion. Despite their plant-dependent life style, aphids were independently rhythmic under light–dark conditions. This first demonstration of diurnal aphid rhythms shows that aphids do not simply track the host plant’s rhythmicity.