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Fig. 2 | Genome Biology

Fig. 2

From: CRISPR guides induce gene silencing in plants in the absence of Cas

Fig. 2

Cas13 and GIGS function across plant species and are heritable. a Representative images of tomato leaves following TRV systemic movement and photobleaching induced by GIGS (TRVm-guide) and an antisense transcript (TRVasPDS). TRV expressing a non-targeting guide crRNA (TRVNT-guide) does not induce photobleaching. b Measurements of chlorophyll content from SPAD meter readings for three independent plants. SPAD meter readings were taken from leaf sections showing photobleaching, and individual reading are shown as black points with the mean and standard deviation shown as a bar plot. c qPCR measurement of the PDS transcript standardized to the EF1α transcript and relative to the NT-guide sample. Three independent samples were analyzed, and individual data are shown as black points with the mean and standard deviation shown as bar plots. d–i Data for independent transgenic Arabidopsis lines. Data for plants expressing LbuCas13a are shown in dark blue and plants not expressing the protein are shown in light blue. Control lines expressing a hairpin construct against the TTG1 transcript are shown in gray. d Trichome counts from the seventh leaf of T2 Arabidopsis lines. Ten plants were counted per independent line, listed below graph, with the individual counts shown as black points and the distribution represented as a violin plot. e Leaf anthocyanin quantification from T3 seedlings following sucrose treatment. Representative wells following extraction shown below each bar plot. f Representative plantlets following sucrose treatment showing anthocyanin pigmentation (i.e., purple color). g Total flavonoids extracted from seeds collected from T2 plants. Five independent seed lots were analyzed per line, shown as black points. h Seed proanthocyanidin quantification from the same plants analyzed in g. i Quantification of the TTG1 transcript from three T2 and three T3 plants per line, individual data shown as black points. d,e,g–i Statistical comparisons were made between the transformation control (no guide) and each treatment using a one-sided Mann-Whitney U test with Benjamini-Hochberg (BH) multiple testing correction. A line is shown above each pair of bars to indicate each pairwise comparison that had a statistically significant difference. This is indicated for comparisons where the adjusted p value was less than 0.05 (*) and 0.01 (**)

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