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Tobacco Hornworm (Manduca Sexta) Caterpillarhemolymph Modulates Reactive Oxygen Species and Calcium Generation in Tomato Protoplasts Akanksha Gandhi, Rupesh Kariyat, Nirakar Sahoo

BIOPHYSICAL JOURNAL(2021)

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摘要
Plants have been at war with herbivorous insects for millions of years and have developed a set of highly regulated defense strategies to sense herbivore attack using chemical cues known as herbivore-associated elicitors (HAEs), including oral secretions, ovipositional fluids, and frass. These HAEs induce a series of signaling cascades, which ultimately provide induced defenses against them. Despite the existing HAEs and their role in plant defense induction, our knowledge of other herbivore generated HAEs in plant-herbivore interactions are limited. in this study, we demonstrate that “hemolymph” from Manduca sexta, caterpillarsalso induce plant defense signaling cascade and thereby act as an HAE.Using a dye-based imaging technique, our study showed that the application of crude M. sexta hemolymph potently increased Reactive Oxygen Species (ROS) productionin isolated tomato protoplasts. The addition of antioxidant NAC (N-acetyl-L-cysteine) antagonized hemolymph-induced ROS generation, indicating that M. sexta hemolymph is a ROS inducer in isolated protoplasts. Furthermore, incubating the protoplasts with Calcium (Ca2+)chelator, BAPTA-AM efficiently abolished the hemolymph-induced ROS production, suggesting possible crosstalk between Ca2+ and ROS signaling. interestingly, the application of crude M. sexta hemolymph dramatically increased Ca2+ in tomato protoplasts. Also, hemolymph-mediated ROS and Ca2+ increase was inhibited in the absence of extracellular Ca2+. Taken together, our study demonstrates that “hemolymph” from Manduca sexta can directly modulate intracellular ROS and Ca2+ production and possibly regulate defenses against insect herbivores by acting as an HAE.
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