Variability in RNA-interference (RNAi) efficacy among different insect orders poses a big hurdle in the development of RNAi-based pest control strategies. The activity of double-stranded ribonucleases (dsRNases) in the digestive canal of insects can be one of the critical factors affecting oral RNAi efficacy. Here, the involvement of these dsRNases in the southern green stinkbug Nezara viridula was investigated. First, the full sequence of the only dsRNase (NvdsRNase) in the transcriptome of N. viridula was obtained, followed by an oral feeding bioassay to evaluate the effect of NvdsRNase-silencing on oral RNAi efficacy. The NvdsRNase was first silenced in nymphs by NvdsRNase-dsRNA injections, followed by exposure to an artificial diet containing a lethal αCop-specific dsRNA. A significantly higher mortality was observed in the NvdsRNase-silenced nymphs when placed on the dsαCop-containing diet (65%) than in the dsGFP injected and dsαCop fed control (46.67%). Additionally, an ex vivo dsRNA degradation assay showed a higher stability of dsRNA in the saliva and midgut juice of NvdsRNase-silenced adults. These results provide evidence for the involvement of NvdsRNase in the reduction of oral RNAi efficacy in N. viridula. This information will be useful in further improving potential RNAi-based strategies to control this pest.

Burgeoning the human population with its required food demand created a burden on ever-decreasing cultivated land and our food production systems. This situation prompted plant scientists to breed crops in a short duration with specific traits. Marker-assisted selection (MAS) has emerged as a potential tool to achieve desirable results in plants with the help of molecular markers and improves the traits of interest in a short duration. The MAS has comprehensively been used in plant breeding to characterize germplasm, diversity analysis, trait stacking, gene pyramiding, multi-trait introgression, and genetic purity of different cereals, pulses, oilseeds, and fiber crops, etc. Mapping studies pointed out several marker-trait associations from different crop species, which specifies the potential application of MAS in accelerating crop improvement. This chapter presents an overview of molecular markers, their genesis, and potential use in plant breeding.