Bacteria of the genus Bacillus can colonize endophytically and benefit several crops including the control of some pest orders. In view of the benefits provided by these microorganisms and in order to find out an efficient biotechnological control for the giant borer, our interest in studying the microorganisms in symbiosis with sugarcane and the giant borer has arisen, since there is no efficient chemical or biological control method for this pest. Therefore, endophytic Bacillus strains were isolated from three sugarcane niches (apoplast fluid, central internode cylinder and roots) and also from the giant borer larvae living inside sugarcane varieties grown in the Northeast region of Brazil. The taxonomical characterization (16S rRNA) of 157 Gram-positive isolates showed that 138 strains belonged to the Bacillus genus. The most representative species were phylogenetically closely related to B. megaterium (11.5%) followed by B. safensis (10.8%), B. cereus (8.9%), B. oleronius (8.9%), B. amyloliquefaciens (7.0%), and B. pacificus (6.4%). BOX-PCR analyses showed very distinct band pattern profiles suggesting a great diversity of Bacillus species within the sugarcane niches and the digestive tract, while the B. cereus group remained very closely clustered in the dendrogram. According to XRE biomarker analysis, eleven strains (FORCN005, 007, 008, 011, 012, 014, 067, 076, 092, 093, and 135) correspond to B. thuringiensis species. Additional studies using conserved genes (glp, gmk, pta, and tpi) indicated that most of these strains were phylogenetically closely related to B. thuringiensis and may be considered different subspecies. In conclusion, this study suggests that the culturable Bacillus species are greatly diversified within the plant niches and showed Bacillus species in the digestive tract of the giant borer for the first time. These results open new perspectives to understand the role and functions played by these microorganisms in symbiosis with this pest and also the possibility of developing an efficient biological control method for the giant borer using strains identified as the B. thuringiensis species.
Planococcus citri (Risso, 1813) (Hemiptera: Pseudococcidae) is an important plant virus vector in grapevine crops in Brazil and other countries. The mealybug grows in roots and leaves of the grapes. Entomopathogenic nematodes (EPNs) are efficient control agents against insects associated to the soil and could be applied with the same equipment used for chemical insecticides. The aim of this study was to select effective EPNs for controlling P. citri females in laboratory conditions (25±1°C, UR 60±10%). We tested 17 native [Steinernema rarum (6 strains), Steinernema glaseri, Steinernema feltiae, Steinernema riobrave, Steinernema sp., Heterorhabditis bacteriophora (7 strains)] and only one exotic strain (Steinernema carpocapsae ALL). The bioassays were done on Petri dishes infested with females of P. citri, which were sprayed with EPNs juveniles. The strain with larger pathogenicity and virulence in laboratory was H. bacteriophora RS33 (from 69.0% to 92.2% of mortality), native of Rio Grande do Sul.
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