Abstract
Main conclusion
ΔClnps6 induced iron redistribution in maize B73 leaf cells and resulted in reactive oxygen species (ROS) burst to enhance plant resistance against Curvularia lunata.
Abstract
Iron is an indispensable co-factor of various crucial enzymes that are involved in cellular metabolic processes and energy metabolism in eukaryotes. For this reason, plants and pathogens compete for iron to maintain their iron homeostasis, respectively. In our previous study, ΔClnps6, the extracellular siderophore biosynthesis deletion mutant of Curvularia lunata, was sensitive to exogenous hydrogen peroxide and virulence reduction. However, the mechanism was not studied. Here, we report that maize B73 displayed highly resistance to ΔClnps6. The plants recruited more iron at cell wall appositions (CWAs) to cause ROS bursts. Intracellular iron deficiency induced by iron redistribution originated form up-regulated expression of genes involved in intracellular iron consumption in leaves and absorption in roots. The RNA-sequencing data also showed that the expression of respiratory burst oxidase homologue (ZmRBOH4) and NADP-dependent malic enzyme 4 (ZmNADP-ME4) involved in ROS production was up-regulated in maize B73 after ΔClnps6 infection. Simultaneously, jasmonic acid (JA) biosynthesis genes lipoxygenase (ZmLOX), allene oxide synthase (ZmAOS), GA degradation gene gibberellin 2-beta-dioxygenase (ZmGA2OX6) and ABA degradation genes abscisic acid hydroxylase (ZmABH1, ZmABH2) involved in iron homeostasis were up-regulated expression. Ferritin1 (ZmFER1) positive regulated maize resistance against C. lunata via ROS burst under Fe-limiting conditions. Overall, our results showed that iron played vital roles in activating maize resistance in B73-C. lunata interaction.
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Abbreviations
- AOS:
-
Allene oxide synthase
- BTS:
-
E3 ligase BRUTUS
- CWAs:
-
Cell wall appositions
- DAB:
-
3,3-Diaminobenzidine
- DCF:
-
CM-H2DCFDA
- DPI:
-
Diphenyleneiodonium chloride
- FER1:
-
Ferritin1
- hpi:
-
Hours post inoculation
- JA:
-
Jasmonic acid
- LOX:
-
Lipoxygenase
- RBOH4:
-
Respiratory burst oxidase homologue
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Acknowledgements
This study was funded by the National Natural Science Foundation of China (Grant No. 31271992). We thank Prof. Junbo Luan (College of Plant Protection, Shenyang Agricultural University), Prof. Xiaofeng Zhu (College of Plant Protection, Shenyang Agricultural University), Prof. Xiaori Han (College of Land and Environment, Shenyang Agricultural University) for instruments support. We would like to thank Plant Bio Press Editing (http://www.plantbiopressediting.com/) for its linguistic assistance during the preparation of this manuscript.
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425_2022_3963_MOESM7_ESM.pdf
Supplementary Fig. S1 Extracellular siderophore biosynthesis analysis of ΔClnps6 and CX-3. a Extracellular siderophore detection on chromeazurol S (CAS) media. The orange halo on CAS media of ΔClnps6 was smaller than that of CX-3, indicating that there was less extracellular siderophore in ΔClnps6. b ΔClnps6 and CX-3 were incubated in MM-Fe media. c HPLC analysis of MM-Fe broth. Extracellular siderophore was extracted and analyzed by HPLC as previously described by Oide et al. (2006). HPLC analysis of each sample of the same proportion of the total extract with a 50 μL aliquot. (PDF 1554 kb)
425_2022_3963_MOESM8_ESM.pdf
Supplementary Fig. S2 Iron accumulation and ROS burst in onion epidermis caused by ΔClnps6 and CX-3. a The accumulation of ferric iron (Fe3+) in onion epidermal cells inoculated by ΔClnps6. b Iron chelator deferoxamine (DFO, 1 mM) was used as control. c H2O2 accumulation in inoculated epidermal cells significantly increased after ΔClnps6 treatment at 24 hpi. (PDF 5948 kb)
425_2022_3963_MOESM9_ESM.pdf
Supplementary Fig. S3 Differentially expressed genes (DEGs) in maize leaves inoculated by ΔClnps6 and CX-3. a, b Venn diagram of co-expression genes in maize leaves inoculated by ΔClnps6 and CX-3 at 6, 24, 48 hpi. c, d The most enriched GO of up-regulated and down-regulated expression in maize leaves inoculated by ΔClnps6 and CX-3. DEGs were filtered with a cut off of log2-fold change ≥ 1 and FDR-corrected P ≤ 0.01. (PDF 2081 kb)
425_2022_3963_MOESM10_ESM.pdf
Supplementary Fig. S4 The most enriched KEGG pathways of up-regulated and down-regulated expression in maize leaves inoculated by ΔClnps6 and CX-3. a-c The up-regulated expression of genes at 6 hpi were mostly associated with glycolysis/gluconeogenesis and oxidative phosphorylation pathway, at 24 hpi with ribosome and protein processing in endoplasmic reticulum pathway, at 48 hpi with plant-pathogen interaction and plant hormone signal transduction pathway. d-f The down-regulated expression of genes at 6 hpi were mainly associated with glycerophospholipid metabolism and plant-pathogen interaction, at 24 hpi with glutathione metabolism, phenylpropanoid biosynthesis and phenylalanine pathway, and at 48 hpi with monoterpenoid biosynthesis and phenylpropanoid biosynthesis pathway. “Rich factor” indicates the ratio of the DEGs number vs the number of genes annotated in this pathway. (PDF 2394 kb)
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Fu, D., Li, J., Yang, X. et al. Iron redistribution induces oxidative burst and resistance in maize against Curvularia lunata. Planta 256, 46 (2022). https://doi.org/10.1007/s00425-022-03963-7
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DOI: https://doi.org/10.1007/s00425-022-03963-7