Bacillus thuringiensis (Bt) is the most widely used biopesticide worldwide and can produce several insecticidal crystal proteins and vegetative insecticidal proteins (Vips) at different growth stages. In our previous study, extracellular polysaccharides (EPSs) of Bt strain HD270 were found to enhance the insecticidal activity of Cry1Ac protoxin against Plutella xylostella (L.) and promote the binding of Cry1Ac to the intestinal brush border membrane vesicles (BBMVs). Whether the synergistic activity of Bt EPSs is common to other Cry1-type or Vip proteins is unclear, as is the potential synergistic mechanism. In this study, crude EPS-HD270 was found to increase the toxicity of Cry1-type toxins and Vip3Aa11 against different lepidopteran pests by approximately 2-fold. The purified EPS-HD270 also possessed synergistic activity against the toxicity of Cry1Ac and Vip3Aa11 against Spodoptera frugiperda (J.E. Smith) and Helicoverpa armigera (Hübner). Furthermore, we found that EPS-HD270 had a strong binding ability with Vip3Aa11 and promoted the binding of Vip3Aa11 to the BBMVs of H. armigera and S. frugiperda. Bt EPS-HD270 also protected Vip3Aa11 from proteolytic processing in larval midgut juice. Bt EPSs had universal synergistic effects on Cry1-type or Vip toxins against S. frugiperda and H. armigera. Bt EPS-HD270 exhibited synergistic activity with Vip3Aa through promotion of binding to BBMVs and protection from digestion by midgut protease. The results indicated that synergistic activity with Bt toxins was an important function of Bt EPSs, which was very different from other Bacillus spp.

Exopolysaccharides (EPSs) are carbohydrate polymers that are synthesized and secreted into the extracellular during the growth of microorganisms. Bacillus thuringiensis (Bt) is a type of entomopathogenic bacterium, that produces various insecticidal proteins and EPSs. In our previous study, the EPSs produced by Bt strains were first found to enhance the toxicity of insecticidal crystal proteins against Plutella xylostella. However, the response of the intestinal bacterial communities of P. xylostella under the action of EPSs is still unelucidated. In this study, 16S rRNA amplicon sequencing was used to characterize the intestinal bacterial communities in P. xylostella treated with EPSs alone, Cry1Ac protoxin alone, and both the Cry1Ac protoxin and EPSs. Compared with the control group, alpha diversity indices, the Chao1 and ACE indices were significantly altered after treatment with EPSs alone, and no significant difference was observed between the groups treated with Cry1Ac protoxin alone and Cry1Ac protoxin + EPSs. However, compared with the gut bacterial community feeding on Cry1Ac protoxin alone, the relative abundance of 31 genera was significantly changed in the group treated with Cry1Ac protoxin and EPSs. The intestinal bacteria, through the oral of Cry1Ac protoxin and EPSs, significantly enhanced the toxicity of the Cry1Ac protoxin towards the axenic P. xylostella. In addition, the relative abundance of the 16S rRNA gene in the chloroplasts of Brassica campestris decreased after adding EPSs. Taken together, these results show the vital contribution of the gut microbiota to the Bt strain-killing activity, providing new insights into the mechanism of the synergistic insecticidal activity of Bt proteins and EPSs.

Rhizosphere bacteria play a crucial role in promoting plant health and development. A full understanding of the bacterial communities in rhizosphere soil and their relationship with those in bulk soil is important for supporting plant growth. Some beneficial bacteria are recruited to the root-zone when plants experience different stresses. However, it is unknown whether the rhizosphere of pear trees enriches some beneficial microorganisms that can resist pathogen infections in natural ecosystems. In this study, we found a higher proportion of antagonistic strains in the rhizosphere of pear trees compared to bulk soil under natural growth conditions. By deep sequencing the V3 and V4 hypervariable regions of 16S rRNA genes, we characterized the bacterial communities in the rhizosphere soil of ‘Yuluxiang’ pear trees and the adjacent bulk soil. Our analysis revealed that the diversity of the bacterial community in the rhizosphere soil was lower than that in the bulk soil, but their compositions differed between the two soil types. We found that 12 phyla, 97 families and 130 genera contributed to these differences. Proteobacteria was the dominant phylum and its relative abundance in the rhizosphere soil was significantly higher than that in the bulk soil. Notably, the two genera, Bacillus and Pseudomonas , were more abundant in the rhizosphere soil. Compared with the bulk soil, the pear rhizosphere soil existed a higher proportion of beneficial bacteria with antagonistic activities against Fusarium oxysporum . These findings indicated that the pear tree rhizosphere can selectively assemble beneficial bacteria with specific antagonistic activities to address threats from pathogens. The distinct bacterial community structure in the rhizosphere of plants reflects a spontaneous ecological adaptation mechanism of plants to their environment.