Lotononis angolensis forms nitrogen fixing, lupinoid nodules with phylogenetically unique, fast-growing, pink-pigmented bacteria, which do not nodulate L. bainesii or L. listii

Novel Burkholderia bacteria isolated from Lebeckia ambigua – A perennial suffrutescent legume of the fynbos

“…S1) and returned to Australia for isolation. Bacteria were isolated from surface sterilised root nodules according to the procedure described by Yates et al (2007).…”

“…Three seedlings, representing each provenance, were placed in a 1 kg pot then inoculated with a single test strain as described by Yates et al (2007). All treatments, plus uninoculated and nitrogen-supplied controls, were replicated four times in this split-plot design, and pots arranged in completely randomised blocks.…”

“…All treatments, plus uninoculated and nitrogen-supplied controls, were replicated four times in this split-plot design, and pots arranged in completely randomised blocks. The split-plot design allows the inclusion of more than one plant genotype in each pot which contains a single inoculant (Howieson et al, 1995;Yates et al, 2007). Sterile DI water was provided every second day and 20 ml of nutrient solution (Howieson et al, 1995) weekly through a sterilised watering tube inserted below the soil surface.…”

“…Nodules were fixed overnight at 4 °C in 3% (v/v) gluteraldehyde in 25 mM phosphate buffer (pH 7.0), then procedures followed as described by Yates et al (2007). The specimens were examined under an Olympus BX51 compound microscope and photographed with an Olympus DP70 digital camera.…”

“…We have collected seed and nodules of several genera of deep-rooted, suffrutescent, perennial legumes (e.g.Rhynchosia, Lebeckia and Lessertia), which although known to botanists (Boatwright et al, 2009;Van Wyk, 1991), are mostly new to agriculture. Our approach has focussed on assessing the nodule bacteria of these potential new agricultural legumes (Ardley et al, 2012;Garau et al, 2009;Gerding et al, 2012;Yates et al, 2007) because poorly adapted inoculants have historically restricted the success of new forage legumes introduced to WA soils (Parker, 1962). Several of the perennial legumes from southern Africa are nodulated by the relatively novel species of Microvirga (Ardley et al, 2012) or by BetaproteobacterialBurkholderia spp.…”