De novo transcriptome analysis and identification of reproduction control genes from the red palm weevil Rhynchophorus ferrugineus

Recent attacks by the red palm weevil, Rhynchophorus ferrugineus (Olivier), have become a severe problem for palm species. In present work, fat body transcriptome of adult female red palm weevil was analyzed, focusing on the identification of reproduction control genes. Transcriptome study was completed by means of next-generation sequencing (NGS) using Illumina Hiseq 2000 sequencing system. A total of 105,938,182 raw reads, 102,645,544 clean reads, and 9,238,098,960 clean nucleotides with a guanine-cytosine content of 40.31%, were produced. The processed transcriptome data resulted in 43,789 unique transcripts (with mean lengths of 1,172 bp). It was found that 20% of total unique transcripts shared up to 80%-100% sequence identity with homologous species, mainly the mountain pine beetle Dendroctonus ponderosae (59.9%) and red flour beetle Tribolium castaneum (26.9%). Nearly 25 annotated genes were predicted to be involved in red palm weevil reproduction, including five vitellogenin (Vg) transcripts. Among the five Vg gene transcripts, one was highly expressed compared with the other four (FPKM values of 1.963, 1.471, 1.028, and 1.017, respectively), and the five Vg gene transcripts were designated as RfVg, RfVg-equivalent1, RfVg-equivalent2, RfVg-equivalent3, and RfVg-equivalent4, respectively. The high expression level of RfVg verified by RT-polymerase chain reaction analysis suggested that RfVg is the primary functional Vg gene in red palm weevil. A high similarity of RfVg with other Coleopterans was also reflected in a phylogenetic tree, where RfVg was placed within the clade of the order Coleoptera. Awareness of the major genes that play critical roles in reproduction and proliferation of red palm weevil is valuable to understand their reproduction mechanism at a molecular level. In addition, for future molecular studies, the NGS dataset obtained will be useful and will promote the exploration of biotech-based control strategies against red palm weevil, a primary pest of palm trees.