Characterization of CRISPR-Cas Systems in Serratia marcescens Isolated from Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae)

Abstract

The CRISPR-Cas adaptive immune system has been attracting increasing scientific interest for biological functions and biotechnological applications. Data on the Serratia marcescens system are scarce. Here, we report a comprehensive characterisation of CRISPR-Cas systems identified in S. marcescens strains isolated as secondary symbionts of Rhynchophorus ferrugineus, also known as Red Palm Weevil (RPW), one of the most invasive pests of major cultivated palms. Whole genome sequencing was performed on four strains (S1, S5, S8, and S13), which were isolated from the reproductive apparatus of RPWs. Subtypes I-F and I-E were harboured by S5 and S8, respectively. No CRISPR-Cas system was detected in S1 or S13. Two CRISPR arrays (4 and 51 spacers) were detected in S5 and three arrays (11, 31, and 30 spacers) were detected in S8. The CRISPR-Cas systems were located in the genomic region spanning from ybhR to phnP, as if this were the only region where CRISPR-Cas loci were acquired. This was confirmed by analyzing the S. marcescens complete genomes available in the NCBI database. This region defines a genomic hotspot for horizontally acquired genes and/or CRISPR-Cas systems. This study also supplies the first identification of subtype I-E in S. marcescens.

Perspectives for Synergic Blends of Attractive Sources in South American Palm Weevil Mass Trapping: Waiting for the Red Palm Weevil Brazil Invasion

“…Given that Rhynchophorus has a palm borer lifestyle, the damage is based on larvae survivorship in the host palm trunk coupled with late damage and infestation diagnosis [ 17 ]. Thus, understanding the insect’s behavior is important for the implementation of new control strategies, especially those derived from research on telemetry [ 37 ], semiochemical interference [ 38 , 39 , 40 , 41 ], new secondary metabolites and formulations [ 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 ], early infestation detection [ 50 ], novel biopesticides [ 51 , 52 , 53 , 54 ], and natural enemies [ 55 , 56 ].…”

“…Rynchophorus ferrugineus completes its life cycle in palms in approximately 80 days; the pest may develop up to four broods per year, with up to 3 months of adult longevity. All the instars live associated with bacteria and other organisms [ 53 , 54 , 85 , 86 , 87 ]. The holometabolic life cycle consists of the expected sequence of egg, larvae, pupae, and adult [ 9 , 28 ] ( Table 1 ).…”

“…Injuries become eventually visible only long after the palm has become infested. Unlike the infestation of R. palmarum that causes the red ring by B. coccophilus , the meristematic tissues of P. canariensis and P. dactylifera suffer infections due to symbiotic bacteria [ 53 , 54 ] and fungi that the RPW females lay by contaminated eggs.…”

A Possible Outbreak by Serratia marcescens: Genetic Relatedness between Clinical and Environmental Strains

“…Genomic restrictions were performed as previously reported [ 13 ] and according to the standard operating procedure for PulseNet PFGE of Escherichia coli O157:H7, E. coli non-O157 (STEC), Salmonella serotypes, Shigella sonnei , and Shigella flexneri [ 14 ]. Agarose-embedded DNA was digested with 40 U Xba I for 4 h at 37 °C.…”

Efficient Genome Editing in Bacillus licheniformis Mediated by a Conditional CRISPR/Cas9 System

“…The clustered regularly interspaced short palindromic repeat (CRISPR) system has been identified as a widespread immune system component in a wide variety of species [11][12][13]. The CRISPR-associated (Cas) proteins, in addition to CRISPR RNA (crRNA) and transactivating CRISPR RNA (tracrRNA), can recognize and introduce double-strand breaks (DSBs) in foreign nucleic acids with the aid of a protospacer-adjacent motif (PAM) sequence, which is the spacer region in the single guide RNA (sgRNA) that is highly specific for the targeted gene [14,15].…”