Gut Microbiota Accelerate the Insecticidal Activity of Plastid-Expressed Bacillus thuringiensis Cry3Bb to a Leaf Beetle, Plagiodera versicolora

Abstract

The contribution of gut microbiota to Bacillus thuringiensis Cry3Bb insecticidal activity in a leaf beetle was demonstrated using transplastomic poplar plants, providing a potential new approach to improve the efficiency of plastid transformation technology for pest control by expression of Bt toxins. Bacillus thuringiensis is widely used as a biopesticide, and its crystal protein expressed in transgenic crops has been successfully used for the management of insect pests. However, whether the midgut microbiota contribute to the Bt insecticidal mechanism remains controversial. We previously demonstrated that transplastomic poplar plants expressing Bt Cry3Bb are highly lethal to willow leaf beetle ( Plagiodera versicolora ), one of the major pests causing severe damage to Salicaceae plants such as willows and poplars. Here, we demonstrate that feeding poplar leaves expressing Cry3Bb to nonaxenic P. versicolora larvae leads to significantly accelerated mortality, and overgrowth and dysbiosis of the gut microbiota, compared with axenic larvae. Corroborating work done with Lepidopteran insects, plastid-expressed Cry3Bb can cause the lysis of the beetle's intestinal cells, lead to the entry of intestinal bacteria into the body cavity, and thus cause the dynamic changes in the flora of the midgut and blood cavity in P. versicolora . Reintroduction of Pseudomonas putida , a gut bacterium of P. versicolora , into axenic P. versicolora larvae further enhances mortality upon feeding on Cry3Bb-expressing poplar. Our results indicate the important contribution of host gut microbiota in promoting the B. thuringiensis crystal protein insecticidal activity and provide new insights into the mechanism of pest control by Bt -transplastomic approaches. IMPORTANCE The contribution of gut microbiota to Bacillus thuringiensis Cry3Bb insecticidal activity in a leaf beetle was demonstrated using transplastomic poplar plants, providing a potential new approach to improve the efficiency of plastid transformation technology for pest control by expression of Bt toxins.