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. 2017 Jul 20;6:e29107. doi: 10.7554/eLife.29107

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© 2017, Keymer et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 2. — Promoter activity indicated by nuclear localized yellow fluorescence in colonized transgenic L. japonicus wild-type roots transformed with constructs containing a 1.5 kb promoter fragment of DIS (A) or a 2.275 kb promoter fragment of RAM2 (B) fused to NLS-YFP. (A-B) Red fluorescence resulting from expression of pSbtM1:SP-mCherry labels the apoplastic space surrounding pre-penetration apparatuus (PPAs) and fungal structures, thereby evidencing the silhouette of these structures. a Colonized root, b non-colonized part of colonized root, c PPAs, (white arrow heads indicate the silhouette of fungal intraradical hyphae) d small arbuscules, e fully developed arbuscules f collapsed arbuscules. Merged confocal and bright field images of whole mount roots are shown. (C-D) Transgenic complementation of dis-1 (C) and ram2-1 (D) hairy roots with the respective wild-type gene driven by the PT4 promoter. The mutant gene was used as negative control. White arrowheads indicate arbuscules. (E-F) Quantification of AM colonization in transgenic roots shown in (C-D). Different letters indicate significant differences (ANOVA; posthoc Tukey; n = 15; p≤0.001) among genotypes for each fungal structure separately. Int. hyphae, intraradical hyphae. (E): F_2,12 = 26.53 (total), F_2,12 = 46.97 (arbuscules), F_2,12 = 27.42 (vesicles). (F) F_2,12 = 341.5 (total), F_2,12 = 146.3 (arbuscules), F_2,12 = 35.86 (vesicles).

DOI: http://dx.doi.org/10.7554/eLife.29107.008

Figure 2—figure supplement 1. — Promoter activity indicated by nuclear localized yellow fluorescence in colonized transgenic L. japonicus wild-type roots transformed with constructs containing a 1.5 kb promoter fragment of DIS (A) or a 2.275 kb promoter fragment of RAM2 (B) fused to NLS-YFP. (A-B) Red fluorescence resulting from expression of pSbtM1:SP-mCherry labels the apoplastic space surrounding pre-penetration apparatuus (PPAs) and fungal structures, thereby evidencing the silhouette of these structures. a Colonized root, b non-colonized part of colonized root, c PPAs, (white arrow heads indicate the silhouette of fungal intraradical hyphae) d small arbuscules, e fully developed arbuscules f collapsed arbuscules. Merged confocal and bright field images of whole mount roots are shown. (C-D) Transgenic complementation of dis-1 (C) and ram2-1 (D) hairy roots with the respective wild-type gene driven by the PT4 promoter. The mutant gene was used as negative control. White arrowheads indicate arbuscules. (E-F) Quantification of AM colonization in transgenic roots shown in (C-D). Different letters indicate significant differences (ANOVA; posthoc Tukey; n = 15; p≤0.001) among genotypes for each fungal structure separately. Int. hyphae, intraradical hyphae. (E): F_2,12 = 26.53 (total), F_2,12 = 46.97 (arbuscules), F_2,12 = 27.42 (vesicles). (F) F_2,12 = 341.5 (total), F_2,12 = 146.3 (arbuscules), F_2,12 = 35.86 (vesicles).

DOI: http://dx.doi.org/10.7554/eLife.29107.008