Environmental response of arbuscular mycorrhizal fungi under soybean cultivation at a regional scale.

Climate change, the shortage of fertilizers and reduced land for cultivation have drawn attention to the potential aid provided by soil-borne organisms. Arbuscular mycorrhizal fungi (AMF) offer a wide range of ecosystem benefits and hence, understanding the mechanisms that control AMF occurrence and maintenance is essential for resilient crop production. We conducted a survey of 123 soybean fields located across a 75,000-km2 area of Argentina to explore AMF community composition and to quantify the impact of soil, climate, and geographical distance on these key soil organisms. First, based upon morphological identification of spores, we compiled a list of the AMF species found in the studied area and identified Acaulospora scrobiculata and Glomus fuegianum as the most frequent species. G. fuegianum abundance was negatively correlated with precipitation seasonality and positively correlated with mean annual precipitation as well as mycorrhizal colonisation of soybean roots. Second, we observed that species richness was negatively correlated with soil P availability (Bray I), clay content and mean annual precipitation. Finally, based on partitioning variation analysis, we found that AMF exhibited spatial patterning at a broad scale. Therefore, we infer that geographical distance was positively associated with spore community composition heterogeneity across the region. Nevertheless, we highlight the importance of precipitation sensitivity of frequent species, overall AMF richness and community composition, revealing a crucial challenge to forthcoming agriculture considering an expected change in global climate patterns.

Reference gene selection for gene expression studies using RT-qPCR in virus-infected planthoppers.

BACKGROUND: Planthoppers not only severely affect crops by causing mechanical damage when feeding but are also vectors of several plant virus species. The analysis of gene expression in persistently infected planthoppers might unveil the molecular basis of viral transmission. Quantitative real-time RT-PCR (RT-qPCR) is currently the most accurate and sensitive method used for quantitative gene expression analysis. In order to normalize the resulting quantitative data, reference genes with constant expression during the experimental procedures are needed. RESULTS: Partial sequences of the commonly used reference genes actin (ACT), α1-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1A), ribosomal protein S18 (RPS18) and polyubiquitin C (UBI) from Delphacodes kuscheli, a planthopper capable of persistently transmitting the plant fijivirus Mal de Río Cuarto virus (MRCV), were isolated for the first time. Specific RT-qPCR primers were designed and the expression stability of these genes was assayed in MRCV-infective and naïve planthoppers using geNorm, Normfinder and BestKeeper tools. The overall analysis showed that UBI, followed by 18S and ACT, are the most suitable genes as internal controls for quantitative gene expression studies in MRCV-infective planthoppers, while TUB and EF1A are the most variable ones. Moreover, EF1A was upregulated by MRCV infection. CONCLUSIONS: A RT-qPCR platform for gene expression analysis in the MRCV-infected planthopper vector Delphacodes kuscheli was developed. Our work is the first report on reference gene selection in virus-infected insects, and might serve as a precedent for future gene expression studies on MRCV and other virus-planthopper pathosystems.