Background: The grasshopper family Lentulidae is endemic to eastern and southern Africa, with its center of diversity situated in South Africa, the highest diversity being found in the Cape Floristic Region, which is one of the global biodiversity hotspots. The family consists of 35 genera sorted in two subfamilies. This study provides first insights into the phylogeny of Lentulidae. Two mitochondrial genes (12S and NDS) were sequenced and the phylogeny was inferred through Maximum Likelihood and Bayesian Inference.
Results: Our results indicate that the current classification into the subfamilies Lentulinae and Shelforditinae may be incorrect as Uvarovidium, Leatettix (Shelforditinae) and Devylderia (Lentulinae) clustered together in one main clade, while Betiscoides, Basutacris and Gymnidium (all Lentulinae) formed the second main clade. The genera Uvarovidium and Leatettix, which had been assigned to the Acrididae (subfamily Hemiacridinae) in the past, grouped within the Lentulidae, confirming their current assignment to this family. The East African Usambilla group is likely to represent a sister clade to the south African Lentula and Eremidium. Diversification patterns in the genus Devylderia and Betiscoides suggest a higher number of species than currently known.
Conclusions: Our phylogeny is not in line with the current systematics of Lentulidae, suggesting that a broader sampling and a study of the genitalia would be useful to clarify the taxonomy. Furthermore, some genera (particularly Betiscoides and Devylderia) are in need of taxonomic revision, as the number of species within these genera is likely to be higher than the current taxonomy suggests.
The third generation of CRISPR/Cas9-mediated genome editing technology has been successfully applied to genome modification of various species including animals, plants and microorganisms. How to improve the efficiency of CRISPR/Cas9 genome editing and reduce its off-target effects has been extensively explored in this field. Using sgRNA (Small guide RNA) with high efficiency and specificity is one of the critical factors for successful genome editing. Several software have been developed for sgRNA design and/or off-target evaluation, which have advantages and disadvantages respectively. In this review, we summarize characters of 16 kinds online and standalone software for sgRNA design and/or off-target evaluation and conduct a comparative analysis of these different kinds of software through developing 38 evaluation indexes. We also summarize 11 experimental approaches for testing genome editing efficiency and off-target effects as well as how to screen highly efficient and specific sgRNA.
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