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Exposure of plants to different biotic and abiotic stress condition instigates significant change in the cellular redox status; resulting in the elevation of reactive nitrogen species that play signaling role in mediating defense responses. Heavy metal associated (HMA) domain containing genes are required for spatio-temporal transportation of metal ions that bind with various enzymes and co-factors within the cell. To uncover the underlying mechanisms mediated by  AtHMA  genes, we identified 14  Arabidopsis HMA  genes that were differentially expressed in response to nitrosative stress through RNA-seq analysis. Of those 14 genes, the expression of eight HMA genes was significantly increased, whereas that of six genes was significantly reduced. We further validated the RNA-seq results through quantitative real-time PCR analysis. Gene ontology analysis revealed the involvement of these genes in biological processes such as hemostasis and transport. The majority of these nitric oxide (NO)-responsive  AtHMA  gene products are carrier/transport proteins.  AtHMAD1  (At1g51090) showed the highest fold change to  S -nitrosocystein. We therefore, further investigated its role in oxidative and nitrosative mediated stress conditions and found that AtHMAD1 has antagonistic role in shoot and root growth. Characterization of  AtHMAD1  through functional genomics showed that the knock out mutant  athmad1  plants were resistant to virulent  Pseudomonas syringae  (DC3000) and showed early induction and high transcript accumulation of pathogenesis related gene. Furthermore, inoculation of  athamd1  with  avirulent  strain of the same bacteria showed negative regulation of  R -gene mediated resistance. These results were supported by hypersensitive cell death response and cell death induced electrolyte leakage. AtHMAD1 was also observed to negatively regulate systemic acquired resistance SAR as the KO mutant showed induction of SAR marker genes. Overall, these results imply that NO-responsive  AtHMA  domain containing genes may play an important role in plant development and immunity.

Nitric Oxide Responsive Heavy Metal-Associated Gene AtHMAD1 Contributes to Development and Disease Resistance in Arabidopsis thaliana