Examining effects of rhizobacteria in relieving abiotic crop stresses using carbon-11 radiotracing

In agriculture, plant growth promoting bacteria (PGPB) are increasingly used for reducing environmental stress-related crop losses through mutualistic actions of these microorganisms, activating physiological and biochemical responses, building tolerances within their hosts. Here we report the use of radioactive carbon-11 (t_½ 20.4 min) to examine the metabolic and physiological responses of Zea mays to Azospirillum brasilense (HM053) inoculation while plants were subjected to salinity and low nitrogen stresses. Host metabolism of "new" carbon resources (as ¹¹ C) and physiology including [¹¹ C]-photosynthate translocation were measured in response to imposed growth conditions. Salinity stress caused shortened, dense root growth with a 6-fold increase in foliar [¹¹ C]-raffinose, a potent osmolyte. ICP-MS analyses revealed increased foliar Na⁺ levels at the expense of K⁺ . HM053 inoculation relieved these effects, reinstating normal root growth, lowering [¹¹ C]-raffinose levels while increasing [¹¹ C]-sucrose and its translocation to the roots. Na⁺ levels remained elevated with inoculation, but K⁺ levels were boosted slightly. Low nitrogen stress yielded longer roots possessing high levels of anthocyanins. Metabolic analysis revealed significant shifts in "new" carbon partitioning into the amino acid pool under low nitrogen stress, with significant increases in foliar [¹¹ C]-glutamate, [¹¹ C]-aspartate, and [¹¹ C]-asparagine, a noted osmoprotectant. ¹¹ CO₂ fixation and [¹¹ C]-photosynthate translocation also decreased, limiting carbon supply to roots. However, starch levels in roots were reduced under nitrogen limitation, suggesting that carbon repartitioning could be a compensatory action to support root growth. Finally, inoculation with HM053 re-instated normal root growth, reduced anthocyanin, boosted root starch, and returned ¹¹ C-allocation levels back to those of unstressed plants.