ALBERT

Description: Targeted genome editing using CRISPR-Cas9 has been widely adopted as a genetic engineering tool in various biological systems. This editing technology has been in the limelight due to its simplicity and versatility compared to other previously known genome editing platforms. Several modifications of this editing system have been established for adoption in a variety of plants, as well as for its improved efficiency and portability, bringing new opportunities for the development of transgene-free improved varieties of economically important crops. This review presents an overview of CRISPR-Cas9 and its application in plant genome editing. A catalog of the current and emerging approaches for the implementation of the system in plants is also presented with details on the existing gaps and limitations. Strategies for the establishment of the CRISPR-Cas9 molecular construct such as the selection of sgRNAs, PAM compatibility, choice of promoters, vector architecture, and multiplexing approaches are emphasized. Progress in the delivery and transgene detection methods, together with optimization approaches for improved on-target efficiency are also detailed in this review. The information laid out here will provide options useful for the effective and efficient exploitation of the system for plant genome editing and will serve as a baseline for further developments of the system. Future combinations and fine-tuning of the known parameters or factors that contribute to the editing efficiency, fidelity, and portability of CRISPR-Cas9 will indeed open avenues for new technological advancements of the system for targeted gene editing in plants.

Description: Fungal endophytes are ubiquitous in nature. They are known as potential sources of natural products, and possible agents for biocontrol attributing to their ability to produce a repertoire of bioactive compounds. In this study, we isolated fungal endophytes from three different tissues (needle, stem and root) of four Pinus species (Pinus densiflora, Pinus koraiensis, Pnus rigida, and Pinus thunbergii) across 18 sampling sites in Korea. A total number of 5872 culturable fungal endophytes were isolated using standard culturing techniques. Molecular identification based on the sequence analyses of the internal transcribed spacer (ITS) or 28S ribosomal DNA revealed a total of 234 different fungal species. The isolated fungal endophytes belonged to Ascomycota (91.06%), Basidiomycota (5.95%) and Mucoromycota (2.97%), with 144 operational taxonomic units (OTUs) and 88 different genera. In all sampling sites, the highest species richness (S) was observed in site 1T (51 OTUs) while the lowest was observed in site 4T (27 OTUs). In terms of diversity, as measured by Shannon diversity index (H’), the sampling site 2D (H′ = 3.216) showed the highest while the lowest H’ was observed in site 2K (H’ = 2.232). Species richness (S) in three different tissues revealed that root and needle tissues are highly colonized with fungal endophytes compared to stem tissue. No significant difference was observed in the diversity of endophytes in three different tissues. Among the four Pinus species, P. thunbergii exhibited the highest species richness and diversity of fungal endophytes. Our findings also revealed that the environmental factors have no significant impact in shaping the composition of the fungal endophytes. Furthermore, FUNGuild analysis revealed three major classifications of fungal endophytes based on trophic modes namely saprotrophs, symbiotrophs, and pathotrophs in four Pinus species, with high proportions of saprotrophs and pathothrops.