Olfactory information drives several behaviours critical for the survival and persistence of insect pests and vectors. Insect behaviour is variable, linked to their biological needs, and regulated by physiological dynamics. For mosquitoes, CO2 is an important cue that signifies the presence of a host, and which elicits activation and attraction. To investigate the genetic basis of olfactory modulation in mosquitoes, we assayed changes in CO2 detection from receptor gene expression through physiological function to behaviour, associated with the onset of host seeking in the malaria vector, Anopheles gambiae. The gene encoding a subunit of the CO2 receptor, AgGr22, was found to be significantly up-regulated in host-seeking females, consistent with a significant increase in sensitivity of CO2-responsive neurons (cpA) housed in capitate peg sensilla of the maxillary palp. In addition, the odorant receptor AgOr28, which is expressed in cpC neurons, was significantly up-regulated. In contrast, AgOr8, which is expressed in cpB neurons, was not affected by this change in physiological state, in agreement with results for the obligate co-receptor Orco. Moreover, the sensitivity of the cpB neuron to (R)-1-octen-3-ol, a well-known mammalian kairomone, did not change in response to the onset of host seeking. The concentration of CO2 flux influenced both the propensity of A. gambiae to take off into the wind and the speed with which this activation occurred. Female A. gambiae mosquitoes responded to CO2 whether mature for host seeking or not, but onset of host seeking enhanced sensitivity and speed of activation at relevant doses of CO2.

Seed systems research is central to achieving the United Nations Sustainable Development Goals. Improved varieties with promise for ending hunger, improving nutrition, and increasing livelihood security may be released, but how do they reach and benefit different types of farmers? Without widespread adoption the genetic gains achieved with improved crop varieties can never be actualized. Progress has been made toward demand responsive breeding, however the draft CGIAR 2030 Research and Innovation Strategy fails to recognize the complexity of seed systems and thus presents a narrow vision for the future of seed systems research. This points to the lack of evidence-based dialogue between seed systems researchers and breeders. This perspective paper presents findings from an interdisciplinary group of more than 50 CGIAR scientists who used a suite of seed systems tools to identify four knowledge gaps and associated insights from work on the seed systems for vegetatively propagated crops (VPCs), focusing on bananas (especially cooking bananas and plantains), cassava, potato, sweetpotato, and yam. We discuss the implications for thinking about and intervening in seed systems using a combined biophysical and socioeconomic perspective and how this can contribute to increased varietal adoption and benefits to farmers. The tools merit wider use, not only for the seed systems of VPCs, but for the seed of crops facing similar adoption challenges. We argue for deeper collaboration between seed systems researchers, breeders and national seed system stakeholders to address these and other knowledge gaps and generate the evidence and innovations needed to break through the 40% adoption ceiling for modern varieties, and ensure good quality seed once the new varieties have been adopted. Without this, the achievements of breeders may remain stuck in the seed delivery pipeline.

BackgroundQuantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures, this technique is subject to systematic error due to biological and technical limitations mainly set by the starting material and downstream procedures. Thus, rigorous data normalization is critical to grant reliability and repeatability of gene expression quantification by qRT-PCR. A number of ‘housekeeping genes’, involved in basic cellular functions, have been commonly used as internal controls for this normalization process. However, these genes could themselves be regulated and must therefore be tested a priori.MethodsWe evaluated eight potential reference genes for their stability as internal controls for RT-qPCR studies of olfactory gene expression in the antennae of Rhodnius prolixus, a Chagas disease vector. The set of genes included were: α-tubulin; β-actin; Glyceraldehyde-3-phosphate dehydrogenase; Eukaryotic initiation factor 1A; Glutathione-S-transferase; Serine protease; Succinate dehydrogenase; and Glucose-6-phosphate dehydrogenase. Five experimental conditions, including changes in age,developmental stage and feeding status were tested in both sexes.ResultsWe show that the evaluation of candidate reference genes is necessary for each combination of sex, tissue and physiological condition analyzed in order to avoid inconsistent results and conclusions. Although, Normfinder and geNorm software yielded different results between males and females, five genes (SDH, Tub, GAPDH, Act and G6PDH) appeared in the first positions in all rankings obtained. By using gene expression data of a single olfactory coreceptor gene as an example, we demonstrated the extent of changes expected using different internal standards.ConclusionsThis work underlines the need for a rigorous selection of internal standards to grant the reliability of normalization processes in qRT-PCR studies. Furthermore, we show that particular physiological or developmental conditions require independent evaluation of a diverse set of potential reference genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0862-x) contains supplementary material, which is available to authorized users.