Fig 2 - uploaded by Junqi Song
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Schematic diagram of the STTM165/166 structure. STTM165/166 consists of tandemly repeated target mimics of miR165/166 separated by a 48-nucleotide imperfect stem-loop linker (48 nt)
Source publication
Virus-based microRNA silencing (VbMS) is an efficient, powerful, and high-throughput approach to screen and investigate the function of microRNAs (miRNAs) in plants. The VbMS system was originally developed in Nicotiana benthamiana and tomato (Solanum lycopersicum) and has been extended to various other plant species such as Arabidopsis, cotton, an...
Context in source publication
Context 1
... The reverse primer consists of a LIC2 linker (5 0 -gAggAgAagA gCCgT-3 0 ), a reverse complement sequence of the TM, and a sequence that is reverse complementary to the 3 0 region of the 48-nt spacer (5 0 -ATTCTTCTTCTTTAGACCAT-3 0 ). For STTM-miR166, the reverse primer is 5 0 -gAggAgAagAgCC gT-TCGGACCAGG-TAG-CTTCATTCCCC-ATTCTTC TTCTTTAGACCAT-3 0 (Fig. ...
Citations
Virus-induced gene silencing (VIGS) is a powerful RNA-silencing technique for transient gene manipulation and functional verification. However, the procedure of its application among dicot and monocot species varies based on types of vectors, plant susceptibility, as well as inoculation methods. Here, we described a simple yet efficient tobacco rattle virus (TRV)-based VIGS system for functional analysis of genes in zoysiagrass (Zoysia japonica cv. “Zenith”), an important warm-season turfgrass species, using Nicotiana benthamiana as an intermediate host. VIGS of the PDS reporter gene resulted in a characteristic photo-bleaching phenotype in majority of the inoculated leaves up to 70% reduction of expression of the endogenous phytoene desaturase (PDS) gene in zoysiagrass. Our protocol provides a fast and efficient toolbox for high-throughput functional genomics in zoysiagrass species, which could potentially be applied to other warm-season turfgrass species.
