McCotter, S. | Search bioRxiv

bioRxiv2026

Toxic Effects of Solanum sisymbriifolium Extracts on Meloidogyne hapla and Meloidogyne chitwoodi

Plant-parasitic nematodes (PPNs) are among the most destructive agricultural pests worldwide, causing significant economic losses across diverse cropping systems. Soil fumigation, the most common management strategy, is costly, detrimental to soil health, and increasingly restricted due to environmental and regulatory concerns. As a result, there is a critical need for alternative, sustainable approaches for PPN control. Solanum sisymbriifolium is resistant to several Meloidogyne species and represents a promising source of natural nematicidal compounds. In this study, we evaluated the effects of S. sisymbriifolium extracts on Meloidogyne chitwoodi and M. hapla, two economically important nematodes. Compounds were extracted using solvents of increasing polarity and then reconstituted in water. The water-solubilized extracts were then used in bioassays to assess their effects on nematode egg hatching, egg viability, and second-stage juvenile (J2) survival. Egg hatching and J2 viability of both Meloidogyne species were consistently affected by compounds in the 1-butanol fraction. Further characterization of these compounds may enable the development of novel, environmentally sustainable alternatives to conventional nematode management strategies.

Plant Biology↗

bioRxiv2019

An Autonomous Molecular Bioluminescent Reporter (AMBER) for voltage imaging in freely moving animals

1.Genetically encoded reporters have greatly increased our understanding of biology, especially in neuroscience. While fluorescent reporters have been widely used, photostability and phototoxicity have hindered their use in long-term experiments. Bioluminescence overcomes some of these challenges but requires the addition of an exogenous luciferin limiting its use. Using a modular approach we have engineered Autonomous Molecular BioluminEscent Reporter (AMBER), an indicator of membrane potential. Unlike other luciferase-luciferin bioluminescent systems, AMBER encodes the genes to express both the luciferase and luciferin. AMBER is a voltage-gated luciferase coupling the functionalities of the Ciona voltage-sensing domain (VSD) and bacterial luciferase, luxAB. When AMBER is co-expressed with the luciferin producing genes it reversibly switches the bioluminescent intensity as a function of membrane potential. Using biophysical and biochemical methods we show that AMBER modulates its enzymatic activity as a function of the membrane potential. AMBER shows several-fold increase in the luminescent ({Delta}L/L) signal upon switching from the off to on state when the cell is depolarized. In vivo expression of AMBER in C. elegans allowed detecting pharyngeal pumping action and mechanosensory neural activity from multiple worms simultaneously. AMBER reports neural activity of multiple animals at the same time and can be used in social behavior assays to elucidate the role of membrane potential underlying behavior. 2. Significance StatementThere have been many exciting advances in the development of genetically encoded voltage indicators to monitor intracelluar voltage changes. Most sensors employ fluorescence, which requires external light, potentially causing photobleaching or overheating. Consequently, there has been interest in developing luminescence reporters. However, they require addition of an exogenous substrate to produce light intracellularly. Here, we engineered a genetically encoded bioluminescent voltage indicator, AMBER, which unlike other bioluminescent activity indicators, does not require addition of an exogenous substrate. AMBER allows a large differential signal, a high signal-to-noise ratio, and causes minimal metabolic demand on cells. We used AMBER to record voltage activity in freely-moving C. elegans, demonstrating that AMBER is a important new tool for monitoring neuronal activity during social behavior.

neuroscience↗