Tobacco Rattle Virus Vector: A Rapid and Transient Means of Silencing Manduca sexta Genes by Plant Mediated RNA Interference

Background: RNAi can be achieved in insect herbivores by feeding them host plants stably transformed to express double stranded RNA (dsRNA) of selected midgut-expressed genes. However, the development of stably transformed plants is a slow and laborious process and here we developed a rapid, reliable and transient method. We used viral vectors to produce dsRNA in the host plant Nicotiana attenuata to transiently silence midgut genes of the plant’s lepidopteran specialist herbivore, Manduca sexta. To compare the efficacy of longer, undiced dsRNA for insect gene silencing, we silenced N. attenuata’s dicer genes (NaDCL1- 4) in all combinations in a plant stably transformed to express dsRNA targeting an insect gene. Methodology/Principal Findings: Stable transgenic N. attenuata plants harboring a 312 bp fragment of MsCYP6B46 in an inverted repeat orientation (ir-CYP6B46) were generated to produce CYP6B46 dsRNA. After consuming these plants, transcripts of CYP6B46 were significantly reduced in M. sexta larval midguts. The same 312 bp cDNA was cloned in an antisense orientation into a TRV vector and Agro-infiltrated into N. attenuata plants. When larvae ingested these plants, similar reductions in CYP6B46 transcripts were observed without reducing transcripts of the most closely related MsCYP6B45. We used this transient method to rapidly silence the expression of two additional midgut-expressed MsCYPs. CYP6B46 transcripts were further reduced in midguts, when the larvae fed on ir-CYP6B46 plants transiently silenced for tw

Knockdown of Midgut Genes by dsRNA-Transgenic Plant-Mediated RNA Interference in the Hemipteran Insect Nilaparvata lugens

BACKGROUND: RNA interference (RNAi) is a powerful technique for functional genomics research in insects. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been reported for lepidopteran and coleopteran insects, showing potential for field-level control of insect pests, but this has not been reported for other insect orders. METHODOLOGY/PRINCIPAL FINDINGS: The Hemipteran insect brown planthopper (Nilaparvata lugens Stål) is a typical phloem sap feeder specific to rice (Oryza sativa L.). To analyze the potential of exploiting RNAi-mediated effects in this insect, we identified genes (Nlsid-1 and Nlaub) encoding proteins that might be involved in the RNAi pathway in N. lugens. Both genes are expressed ubiquitously in nymphs and adult insects. Three genes (the hexose transporter gene NlHT1, the carboxypeptidase gene Nlcar and the trypsin-like serine protease gene Nltry) that are highly expressed in the N. lugens midgut were isolated and used to develop dsRNA constructs for transforming rice. RNA blot analysis showed that the dsRNAs were transcribed and some of them were processed to siRNAs in the transgenic lines. When nymphs were fed on rice plants expressing dsRNA, levels of transcripts of the targeted genes in the midgut were reduced; however, lethal phenotypic effects after dsRNA feeding were not observed. CONCLUSIONS: Our study shows that genes for the RNAi pathway (Nlsid-1 and Nlaub) are present in N. lugens. When insects were fed on rice plant materials expressing dsRNAs, RNA interference was triggered and the target genes transcript levels were suppressed. The gene knockdown technique described here may prove to be a valuable tool for further investigations in N. lugens. The results demonstrate the potential of dsRNA-mediated RNAi for field-level control of planthoppers, but appropriate target genes must be selected when designing the dsRNA-transgenic plants

Rapid Determination of Fibronectin by Laser Nephelometry Fibronectin concentrations in plasma in human diseases, I.

Peer Reviewe

Objective cognitive performance and subjective complaints in patients with chronic Q fever or Q fever fatigue syndrome

Contains fulltext : 219232.pdf (publisher's version ) (Open Access)Background: Primary aim of this study was to compare cognitive performance of patients with chronic Q fever or Q fever fatigue syndrome (QFS) to matched controls from the general population, while taking performance validity into account. Second, we investigated whether objective cognitive performance was related to subjective cognitive complaints or psychological wellbeing. Methods: Cognitive functioning was assessed with a neuropsychological test battery measuring the domains of processing speed, episodic memory, working memory and executive functioning. Tests for performance validity and premorbid intelligence were also included. Validated questionnaires were administered to assess self-reported fatigue, depressive symptoms and cognitive complaints. Results: In total, 30 patients with chronic Q fever, 32 with QFS and 35 controls were included. A high percentage of chronic Q fever patients showed poor performance validity (38%) compared to controls (14%, p = 0.066). After exclusion of participants showing poor performance validity, no significant differences between patients and controls were found in the cognitive domains. QFS patients reported a high level of cognitive complaints compared to controls (41.2 vs 30.4, p = 0.023). Cognitive complaints were not significantly related to cognitive performance in any of the domains for this patient group. Conclusions: The high level of self-reported cognitive complaints in QFS patients does not indicate cognitive impairment. A large proportion of the chronic Q fever patients showed suboptimal mental effort during neuropsychological assessment. More research into the underlying explanations is needed. Our findings stress the importance of assessing cognitive functioning by neuropsychological examination including performance validity, rather than only measuring subjective cognitive complaints.8 p

Agriculture and forest: A sustainable strategy in the Brazilian Amazon.

Large-scale agriculture is increasing in anthropogenically modified areas in the Amazon Basin. Crops such as soybean, maize, oil palm, and others are being introduced to supply the world demand for food and energy. However, the current challenge is to enhance the sustainability of these areas by increasing efficiency of production chains and to improve environmental services. The Amazon Basin has experienced a paradigm shift away from the traditional slash-and-burn agricultural practices, which offers decision makers the opportunity to make innovative interventions to enhance the productivity in previously degraded areas by using trees to ecological advantage. This study describes a successful experiment integrating the production of soybean and paricá (Glycine max L. and Schizolobium amazonicum) based on previous research that indicated potential topoclimatic zones for planting paricá in the Brazilian state of Pará. This paper shows that a no-tillage system reduces the effects of drought compared to conventional tillage still used by many farmers in the region. The integrated system was implemented during the 2014/2015 season in 234.6 ha in the high-potential zone in the municipality of Ulianópolis, Pará. Both soybean and paricá were planted simultaneously. Paricá was planted in 5 m x 2 m inter-tree spacing totaling 228x103 trees per hectare and soybean, in 4 m x 100 m spacing, distributed in nine rows with a 0.45 m inter-row distance, occupying 80% of the area. The harvested soybean production was 3.4 t ha-1, higher than other soybean monocultures in eastern Pará. Paricá benefited from soybean fertilization in the first year: It exhibited rapid development in height (3.26 m) and average diameter (3.85 cm). Trees and crop rotation over the following years is six years for forest species and one year for each crop. Our results confirm there are alternatives to the current production systems able to diminish negative impacts resulting from monoculture. In addition, the system provided environmental services such as reduced soil erosion and increased carbon stock by soil cover with no-tillage soybean cultivation. The soybean cover contributes to increased paricá thermal regulation and lower forestry costs. We concluded that innovative interventions are important to show local farmers that it is possible to adapt an agroforest system to large-scale production, thus changing the Amazon.201

Estimating Relevant Input Dimensions for Self-organizing Algorithms

this paper. Approaches like [11] clearly indicate that often a considerable reduction of the data dimension is possible without loss of informatio