The plant substrate triggers <i>dnf2</i> fix⁻ phenotype at distance.

<p><i>dnf2–4</i> and WT plants nodulated by <i>S. meliloti</i> Rm41 were grown on agarose-based BNM. Agar- or agarose-based plugs (1.5×1×0.5 cm) were laid onto root systems at 15 dpi and the color and numbers of nodules produced by 24 plants were monitored. The experimental set up is illustrated in (a). (b) Distribution of nodule classes 35 days after plug addition for WT and <i>dnf2–4</i> grown on agarose based medium. The experiment was repeated three times with similar results. Statistically identical distribution are attributed identical letters (Chi-Square Test of Homogeneity with Bonferroni correction, p-value = 1.32e-06).</p

Influence of the plant growth conditions on <i>dnf2</i> phenotype is transient.

<p>(a–c) Frequencies of nodule classes after transfer to agar or agarose medium. <i>M. truncatula dnf2–4</i> and WT plants (n = 24 for every conditions) inoculated with <i>S. meliloti</i> Rm41 were cultivated <i>in vitro</i> on BNM using agar or agarose as a gelling agent for 14 days and transfer to new medium with the same or different gelling agent. Pink nodules are represented by diamonds, white nodules by open squares and brownish nodules by triangles. The experiment was repeated three times with similar results. (d) analysis of the distribution of nodule classes at 35 days after transfer. Statistically identical distribution are attributed identical letters (Chi-Square Test of Homogeneity with Bonferroni correction, p-value = 2.2–16).</p

Plant growth conditions impact the <i>dnf2</i> phenotype.

<p>Nodules were harvested 18 days post inoculation with Rm41. a-i, Wild-type nodules. j-r, <i>dnf2–4</i> nodules. Left panels, middle panels and right panels represent nodules grown on agar-, agarose- and agarose-based BNM supplemented with 1% ulvan respectively. Panels a, b, c, j, k and l (scale bars 500 µm) illustrate whole nodules, panels d, e, f, m, n and o (scale bars 200 µm except for d 500 µm) are thin sections of whole nodules and panels g, h, i, p, q, r (scale bars 50 µm) are enlargement of the zone III of panels d, e, f, m, n and o, respectively.</p

Bacteroid differentiation defect is not sufficient to trigger defense-like reactions in <i>dnf2</i> nodules.

<p>Panel a: Expression of defense markers was evaluated by RT-qPCR using cDNA prepared from 14 dpi nodules. The y axis represents fold induction/WT. Panel b: 25 dpi nodules of R108 WT induced by Sm1021 WT. Panel c: 25 dpi nodules of <i>dnf2–4</i> induced by Sm1021 WT. Panel d: 25 dpi nodules of R108 WT induced by a SM1021 <i>bacA</i> derivative. Nodules in b, c and d were stained for phenolics using potassium permanganate toluidine blue (scale bars 500 µm).</p

Ulvan abolishes <i>dnf2</i> nitrogen fixation on permissive condition.

<p><i>M. truncatula</i> WT R108 and <i>dnf2–4</i> plants were cultivated on agarose BNM supplemented or not with 1% ulvan. Acetylene reduction assays were conducted on plants 27 dpi with <i>S. medicae</i> WSM419 (n = 8) (a). <i>M. truncatula</i> WT R108 and <i>dnf2–4</i> plants were cultivated on Phytagel BNM supplemented or not with 10 mM CaSO₄. Acetylene reduction assays were conducted on plants 14 dpi with <i>S. medicae</i> WSM419 (n = 5) (b). A Mann-Whitney test was performed between WT and <i>dnf2-1</i> mutant for each condition. Stars indicate significant differences (** p-value <1e-03) Error bars represent standard errors.</p

Mentha viridis L.

原著和名: ミドリハクカ科名: シソ科 = Labiatae採集地: 千葉県 千葉市 千葉大学 (下総 千葉市 千葉大学)採集日: 1967/8/13採集者: 萩庭丈壽整理番号: JH042898国立科学博物館整理番号: TNS-VS-99289

Additional file 3: Figure S1. of Evaluation of off-target and on-target scoring algorithms and integration into the guide RNA selection tool CRISPOR

Overlap of off-target detection for the EMX1 and VEGFA guides tested by different assays. Off-targets are only shown if they were detected by at least a single study and with a frequency of 0.1 %. See Additional file 1: Tables S1 and Additional file 4: Table S2 for the modification frequencies and additional details on the off-targets for the guides EMX1 and VEGFA, respectively. Additional file 4: Table S2 also includes the data by Hsu et al. [7], who quantified cleavage at putative off-target loci predicted by the CRISPR Design website ( http://crispr.mit.edu/ ) with targeted deep sequencing, Tsai et al. [3], who isolated double-strand breaks with modified oligonucleotides followed by sequencing, Frock et al. [28], who detected translocations, and Kim et al. [33] and Kim et al. [27], who performed whole-genome sequencing to find CRISPR-induced modifications. For details on the different studies, see Additional file 1: Table S1. (PDF 17 kb

Acanthopanax trichodon Franch. et Savat.

原著和名: ミヤマウコギ科名: ウコギ科 = Araliaceae採集地: 千葉県 清澄山 (上総〜安房 清澄山)採集日: 1961/6/18採集者: 萩庭丈壽整理番号: JH042319国立科学博物館整理番号: TNS-VS-99231

Ilex serrata Thunb.

原著和名: ウメモドキ科名: モチノキ科 = Aquifoliaceae採集地: 愛知県 豊橋市 岩崎町 (三河 豊橋市 岩崎)採集日: 1968/10/20採集者: 萩庭丈壽整理番号: JH042317国立科学博物館整理番号: TNS-VS-99231