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

Fig. 2.

Fig. 2.

Symbiotic phenotypes of minSymA2.0 and minSymA2.1 assembled by yeast recombineering. (A) Map of minSymA2.0 and minSymA2.1, with genes represented as arrows. Annotations and coloring are as described in Fig. 1B. (B) Shoot dry weight of alfalfa inoculated with minSymA2.0 in a ΔpSymA background (38 d post inoculation). (C) Shoot dry weight of alfalfa inoculated with the ∆A121 mutant with and without the fix-1 locus assembled by yeast recombineering and integrated into the LP-B1. Shoot dry-weight measurements were taken 28 d post inoculation. (D) Map of pSymA subdeletions (E). Complementation of the ∆A401-∆A406 deletions with minSymA2.0 introduced by transduction (measured 28 d post inoculation). (F) Complementation of the ∆A401 deletion strain with the fix-2 locus and the SDW phenotype of minSymA2.1 (minSymA2.0 + fix-2) (38 d post inoculation). For B, C, E, and F, each data point represents one independent replicate and is calculated from the average shoot dry weight in one pot (six plants per pot). (G) Representative image of M. sativa plants in pots. The labels below each pot indicate the strain used as inoculum. (Scale bar, 2 cm.) (H) Average number of Fix+ nodules formed per plant by minSymA1.0 and minSymA2.1 compared to wild type. Each data point represents one independent replicate calculated from the average number of nodules per plant in one pot (six plants per pot). (I) Kinetics of nodule formation by minSymA2.1 compared to wild type. Data are the average of two independent experiments with 16 plants per experiment. (J) Percentage nodule occupancy of minSymA1.0 and minSymA2.1 when competed as 80% of the inoculum with the wild type. Each data point represents one independent replicate and is calculated from the proportion in one pot (four plants per pot). Shoot dry-weight assays and nodule number statistical significance was assessed with one-way ANOVA and Dunnett’s multiple comparisons tests and is presented compared to wild type. Significance of percentage nodule occupancy was tested with a two-way paired t test. For all significance testing: *P < 0.1, **P < 0.01, ***P < 0.001, ****P < 0.0001.