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. 2020 Dec 31;118(2):e2018015118. doi: 10.1073/pnas.2018015118

Fig. 4.

Fig. 4.

Symbiotic phenotypes of minSymA3.3 modules and chimera with NGR234 nod genes. (A) Schematic showing the locations where the nod, nif, and fix symbiotic clusters were integrated into the S. meliloti genome. Map of the synthetic nod, nif, and fix clusters with genes represented as arrows. Annotations and coloring are as described in Fig. 1B. (B) Shoot dry-weight accumulation by S. meliloti hosts inoculated with an S. meliloti strain with synth-nod, synth-nif, and synth-fix integrated into different locations in the genome. Each data point represents one independent replicate and is calculated from the average shoot dry weight in one pot (six plants per pot). (C) Nodule formation on S. fredii NGR234 hosts by S. meliloti minSymA3.3 engineered with NGR234 nod genes (28 d post inoculation). Each data point represents one independent replicate and is calculated from the average number of nodules per plant in one pot (six plants per pot for M. sativa, four plants per pot for V. unguiculata and M. atropurpureum, and two plants per pot for L. leucocephala). (D) Light microscopy images of representative nodules formed by S. fredii NGR234 and S. meliloti minSymA3.3 engineered with NGR234 nod genes. (Scale bars, 2 mm.) (E) Confocal microscopy of nodules formed by S. fredii NGR234 and S. meliloti minSymA3.3 engineered with NGR234 nod genes. (Scale bars, 250 μm.) For shoot dry-weight assays and nodule number, statistical significance was assessed with one-way ANOVA and Dunnett’s multiple comparisons test and is presented compared to wild-type S. meliloti. *P < 0.1, **P < 0.01, ***P < 0.001, ****P < 0.0001.