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Paraburkholderia species promote Broccoli growth in a cultivar-dependent manner
Root tip inoculation of the two Broccoli cultivars with strains of three different Paraburkholderia species led to changes in leaf color (deep green leaves), shoot biomass, root biomass and root architecture (Fig. 1a). Percent change in biomass was used as a measure to assess the growth-promoting effects of the Paraburkholderia species in the two Broccoli cultivars. Two-way analysis of variance (ANOVA) was conducted to assess the influence of the two independent variables (strains of Paraburkholderia species and Broccoli cultivars) on both shoot and root biomass. The Paraburkholderia species included three levels (Pbg, Pbh, Pbt) and the Broccoli cultivars consisted of two levels (Coronado, Malibu). For shoots, all interactions, except Pbt-Malibu, resulted in significant increases in biomass relative to the non-treated control plants, while for roots all three Paraburkholderia species significantly increased the biomass in both Broccoli cultivars (Fig. 1b). In general, the relative impact of Paraburkholderia species was up to 3 times higher for root biomass than for shoot biomass (Fig. 1b). Two-way ANOVA showed highly significant interactions between the strains of Paraburkholderia species and Broccoli cultivars regarding the percent changes in shoot and root biomass (Supplementary Table S1). Overall, for cultivar Coronado the percent change in shoot biomass was about 40% compared to the control, and not significantly different between the different strains of Paraburkholderia species, whereas in cultivar Malibu the percent change in shoot biomass was significantly higher for Pbg (~ 70%) and Pbh (~ 90%) as compared to Pbt. Furthermore, inoculation with Pbh led to a significantly higher increase in shoot biomass in cultivar Malibu than in Coronado. Regarding the percent change in root biomass, only inoculation of Pbt showed significant differences between the two Broccoli cultivars. As indicated above, the shoot biomass of cultivar Malibu inoculated with Pbt was not significantly different from the control plants (Fig. 1b). Over a period of 11 days, both Pbg and Pbh-treated Broccoli cultivars showed significantly higher shoot and root biomass from 7 days post inoculation (dpi) onwards, while Pbt-treated plants showed higher shoot biomass in Coronado from 9 dpi onwards (Fig. 1c).
Figure 1

Biomass and phenotypic changes in Broccoli cultivars in response to root tip inoculation with strains of three Paraburkholderia species. (a) Pictures of MS agar plate with two Broccoli cultivars (Coronado and Malibu) at 11 days post inoculation with strains of three Paraburkholderia species (Pbg: Paraburkholderia graminis PHS1, Pbh: P. hospita mHSR1, and Pbt: P. terricola mHS1). (b) Percent changes in shoot and root biomass (mean ± standard error, n = 4 (shoot) and n = 6 (root)) of two Broccoli cultivars inoculated with the strains of the Paraburkholderia species. Treatments sharing the same letters are not significantly different (Two-way ANOVA, Tukey’s HSD post hoc test, P  More