Immune response of Phyllophaga polyphylla larvae is not an effective barrier against Metarhizium pingshaense

Abstract

Previous research has uncovered that the cuticle of P. polyphylla larvae acts as a good nonimmunological barrier against M. pingshaense. In the present study we investigated whether P. polyphylla larvae also show a similarly robust immunological response against M. pingshaense. Firstly, we estimated a median lethal dose (LD50) of blastospores to be injected into the hemocoel. Secondly, we injected the estimated LD50 of blastospores into the hemocoel of larvae to quantify phenoloxidase (PO), nitric oxide (NO) and antimicrobial activity as a response against fungal invasion. In contrast to a previous report that showed that M. pingshaense is unable to kill P. polyphylla after topical applications, here we demonstrate that: (a) 100 % of P. polyphylla larvae died when blastospores were injected into the hemocoel and (b) when injecting the LD50 into the hemocoel of the larvae, immune response did not differ with control. Our results imply that immunological responses do not protect P. polyphylla larvae against M. pingshaense infections. Thus, the cuticle seems a better defense mechanism compared to PO, NO and antimicrobial activity. One proximate explanation for our results is that blastospores are not detected by the host’s immune machinery. An ultimate explanation is that there may be a resource-based tradeoff between non-immunological and immunological barriers, in which white grubs may be investing more in cuticle at the cost of PO, NO and antimicrobial activity.