Phenotypic characterization of Arabidopsis thaliana lines overexpressing AVP1 and MIOX4 in response to abiotic stresses.

PREMISE: AVP1 (H+-pyrophosphatase) and MIOX4 (myo-inositol oxygenase) are genes that, when overexpressed individually, enhance the growth and abiotic stress tolerance of Arabidopsis thaliana plants. We propose that pyramiding AVP1 and MIOX4 genes will further improve stress tolerance under water-limited and salt-stress conditions. METHODS: MIOX4 and AVP1 reciprocal crosses were developed and phenomic approaches used to investigate the possible synergy between these genes. RESULTS: Under normal and stress conditions, the crosses had higher foliar ascorbate content than the wild-type and parental lines. Under water-limited conditions, the crosses also displayed an enhanced growth rate and biomass compared with the control. The observed increases in photosystem II efficiency, linear electron flow, and relative chlorophyll content may have contributed to this observed phenotype. Additionally, the crosses retained more water than the controls when subjected to salt stress. Higher seed yields were also observed in the crosses compared with the controls when grown under salt and water-limitation stresses. DISCUSSION: Overall, these results suggest the combinatorial effect of overexpressing MIOX4 and AVP1 may be more advantageous than the individual traits for enhancing stress tolerance and seed yields during crop improvement.

Mechanisms underlying the enhanced biomass and abiotic stress tolerance phenotype of an Arabidopsis MIOX over-expresser.

Myo-inositol oxygenase (MIOX) is the first enzyme in the inositol route to ascorbate (L-ascorbic acid, AsA, vitamin C). We have previously shown that Arabidopsis plants constitutively expressing MIOX have elevated foliar AsA content and displayed enhanced growth rate, biomass accumulation, and increased tolerance to multiple abiotic stresses. In this work, we used a combination of transcriptomics, chromatography, microscopy, and physiological measurements to gain a deeper understanding of the underlying mechanisms mediating the phenotype of the AtMIOX4 line. Transcriptomic analysis revealed increased expression of genes involved in auxin synthesis, hydrolysis, transport, and metabolism, which are supported by elevated auxin levels both in vitro and in vivo, and confirmed by assays demonstrating their effect on epidermal cell elongation in the AtMIOX4 over-expressers. Additionally, we detected up-regulation of transcripts involved in photosynthesis and this was validated by increased efficiency of the photosystem II and proton motive force. We also found increased expression of amylase leading to higher intracellular glucose levels. Multiple gene families conferring plants tolerance/expressed in response to cold, water limitation, and heat stresses were found to be elevated in the AtMIOX4 line. Interestingly, the high AsA plants also displayed up-regulation of transcripts and hormones involved in defense including jasmonates, defensin, glucosinolates, and transcription factors that are known to be important for biotic stress tolerance. These results overall indicate that elevated levels of auxin and glucose, and enhanced photosynthetic efficiency in combination with up-regulation of abiotic stresses response genes underly the higher growth rate and abiotic stresses tolerance phenotype of the AtMIOX4 over-expressers.