Effects of <i>Bacillus</i> spp. inoculation on suggested shoot tolerance mechanisms in lowland rice (<i>Oryza sativa</i> L.) grown under iron toxicity

Abstract Background In areas of lowland rice production, high iron concentrations in the soil often lead to yield reductions. Local adapted varieties possess different adaptation mechanisms, which, however, are not fully understood. Previous studies have shown that endophytic bacteria can influence plant tolerance to abiotic stresses, including iron toxicity. Aim This study aims at analyzing the effects of different Bacillus isolates on distinct shoot tolerance mechanism in different rice cultivars grown under iron toxicity. Methods Three lowland rice cultivars, varying in their tolerance against iron toxicity (IR31785‐58‐1‐2‐3‐3, Sahel 108, Suakoko 8), were inoculated with three Bacillus strains (two of B. pumilus and one of B. megaterium ). One week after Bacillus inoculation plants were subjected to high iron levels (1000 ppm) for 7 days. Leaf symptom scoring was used to assess tolerance levels. Activities of ascorbate peroxidase (APX), glutathione reductase (GR), catalase (CAT), superoxide dismutase (SOD), and guaiacol peroxidase (PRX) were measured by spectrophotometric assays. Transcription of genes related to iron toxicity ( OsFER, OsFRO1, OsNRAMP6 ) was determined by RT‐qPCR. Bacterial production of NO was evaluated by measuring nitrite levels in the culture supernatants. Results In general, iron toxicity affected the activities of APX, GR, CAT, and PRX but not SOD activity. Only PRX activity in response to iron differed between cultivars with a significantly stronger increase in IR31785‐58‐1‐2‐3‐3. Inoculation with B. pumilus Ni9MO12 led to higher activity of CAT in the leaf sheaths of all cultivars and an increase in GR activity in the sheaths that was significantly higher in Suakoko 8. In the young leaf blades of IR31785‐58‐1‐2‐3‐3, transcription of OsFRO1 and OsNRAMP6 was not significantly affected by Bacillus inoculation, whereas accumulation of OsFER mRNA was significantly higher in iron‐stressed, B. pumilus Ni9MO12 inoculated plants compared to non‐inoculated, non‐iron‐stressed plants. Nitrite concentration as an indicator for NO production was increased in B. pumilus Ni9MO12 culture supernatants. Conclusion Our results show that in the sensitive cultivar IR31785‐58‐1‐2‐3‐3 tolerance to iron toxicity increases when inoculated with B. pumilus Ni9MO12, coinciding with higher levels of ferritin transcription. NO production by the Bacillus isolate might confer the promotion of OsFER gene transcription in the inoculated plants.