SUMMARYIntracellular invasion of root cells is required for the establishment of successful endosymbioses in legumes of both arbuscular mycorrhizal (AM) fungi and rhizobial bacteria. In both interactions a requirement for successful entry is the activation of a common signalling pathway that includes five genes required to generate calcium oscillations and two genes required for the perception of the calcium response. Recently, it has been discovered that in Medicago truncatula, the Vapyrin (VPY) gene is essential for the establishment of the arbuscular mycorrhizal symbiosis. Here, we show by analyses of mutants that the same gene is also required for rhizobial colonization and nodulation. VPY encodes a protein featuring a Major Sperm Protein domain, typically featured on proteins involved in membrane trafficking and biogenesis, and a series of ankyrin repeats. Plants mutated in this gene have abnormal rhizobial infection threads and fewer nodules, and in the case of interactions with AM fungi, epidermal penetration defects and aborted arbuscule formation. Calcium spiking in root hairs in response to supplied Nod factors is intact in the vpy-1 mutant. This, and the elevation of VPY transcripts upon application of Nod factors which we show to be dependent on NFP, DMI1, and DMI3, indicates that VPY acts downstream of the common signalling pathway.
Land applications of manure from confined animal systems and direct deposit by grazing animals are both major sources of bacteria in streams. An understanding of the overland transport mechanisms from land applied waste is needed to improve design of best management practices (BMPs) and modeling of nonpoint source (NPS) pollution. Plots were established on pasturelands receiving phosphorus-based livestock waste applications to measure the concentrations of Escherichia coli (E. coli), fecal coliform (FC), and Enterococcus present in overland flow at the edge of the field. The flow-weighted bacteria concentrations were highest in runoff samples from the plots treated with cowpies (1.37 × 10 5 colony forming units (cfu)/100 ml of E. coli) followed by liquid dairy manure (1.84 × 10 4 cfu/100 ml of E. coli) and turkey litter (1.29 × 10 4 cfu/100 ml of E. coli). The temporal distribution of fecal bacterial concentrations appeared to be dependent upon both the animal waste treatment and the indicator species, with peak concentrations occurring either at the beginning of the runoff event or during peak flow rates. BMPs could be selected to reduce peak flows or first flush effects depending upon the litter or manure applied to the land. The commercial Biolog System was used to identify the dominant species of Enterococcus present in the cowpie source manure (Enterococcus mundtii 55%) and in the runoff collected from the transport plots treated with cowpies (Enterococcus faecalis 37%). The identification of predominant species of Enterococcus that are associated with specific sources of fecal pollution could greatly assist with identifying the origins of NPS pollution.
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