Functional interplay between protein kinase CK2 and salicylic acid sustains PIN transcriptional expression and root development.

We have previously reported that CK2-defective Arabidopsis thaliana plants (CK2mut plants) were impaired severely in root development and auxin polar transport, and exhibited transcriptional misregulation of auxin-efflux transporters (Plant J., 67, 2011a, 169). In this work we show that CK2mut roots accumulate high levels of salicylic acid (SA) and that the gene that encodes isochorismate synthase (SID2) is overexpressed, strongly suggesting that CK2 activity is required for SA biosynthesis via the shikimate pathway. Moreover, SA activates transcription of CK2-encoding genes and, thus, SA and CK2 appear to be part of an autoregulatory feed-back loop to fine-tune each other's activities. We also show that exogenous SA and constitutive high SA levels in cpr mutants reproduce the CK2mut root phenotypes (decrease of root length and of number of lateral roots), whereas inhibition of CK2 activity in SA-defective and SA-signalling mutants lead to less severe phenotypes, suggesting that the CK2mut root phenotypes are SA-mediated effects. Moreover, exogenous SA mediates transcriptional repression of most of PIN-FORMED (PIN) genes, which is the opposite effect observed in CK2mut roots. These results prompted us to propose a model in which CK2 acts as a link between SA homeostasis and transcriptional regulation of auxin-efflux transporters. We also show that CK2 overexpression in Arabidopsis has neither impact on SA biosynthesis nor on auxin transport, but it improves the Arabidopsis root system. Thus, unlike the outcome in mammals, an excess of CK2 in plant cells does not produce neoplasia, but it might be advantageous for plant fitness.

Different Cis-Regulatory Elements Control the Tissue-Specific Contribution of Plastid ω-3 Desaturases to Wounding and Hormone Responses.

Trienoic fatty acids are essential constituents of biomembranes and precursors of jasmonates involved in plant defense responses. Two ω-3 desaturases, AtFAD7 and AtFAD8, synthetize trienoic fatty acids in the plastid. Promoter:GUS and mutagenesis analysis was used to identify cis-elements controlling AtFAD7 and AtFAD8 basal expression and their response to hormones or wounding. AtFAD7 promoter GUS activity was much higher than that of AtFAD8 in leaves, with specific AtFAD7 expression in the flower stamen and pistil and root meristem and vasculature. This specific tissue and organ expression of AtFAD7 was controlled by different cis-elements. Thus, promoter deletion and mutagenesis analysis indicated that WRKY proteins might be essential for basal expression of AtFAD7 in leaves. Two MYB target sequences present in the AtFAD7 promoter might be responsible for its expression in the flower stamen and stigma of the pistil and in the root meristem, and for the AtFAD7 wound-specific response. Two MYB target sequences detected in the distal region of the AtFAD8 gene promoter seemed to negatively control AtFAD8 expression, particularly in true leaves and flowers, suggesting that MYB transcription factors act as repressors of AtFAD8 gene basal expression, modulating the different relative abundance of both plastid ω-3 desaturases at the transcriptional level. Our data showed that the two ABA repression sequences detected in the AtFAD7 promoter were functional, suggesting an ABA-dependent mechanism involved in the different regulation of both ω-3 plastid desaturases. These results reveal the implication of different signaling pathways for the concerted regulation of trienoic fatty acid content in Arabidopsis.

Non-redundant Contribution of the Plastidial FAD8 ω-3 Desaturase to Glycerolipid Unsaturation at Different Temperatures in Arabidopsis.

Plastidial ω-3 desaturase FAD7 is a major contributor to trienoic fatty acid biosynthesis in the leaves of Arabidopsis plants. However, the precise contribution of the other plastidial ω-3 desaturase, FAD8, is poorly understood. Fatty acid and lipid analysis of several ω-3 desaturase mutants, including two insertion lines of AtFAD7 and AtFAD8, showed that FAD8 partially compensated the disruption of the AtFAD7 gene at 22 °C, indicating that FAD8 was active at this growth temperature, contrasting to previous observations that circumscribed the FAD8 activity at low temperatures. Our data revealed that FAD8 had a higher selectivity for 18:2 acyl-lipid substrates and a higher preference for lipids other than galactolipids, particularly phosphatidylglycerol, at any of the temperatures studied. Differences in the mechanism controlling AtFAD7 and AtFAD8 gene expression at different temperatures were also detected. Confocal microscopy and biochemical analysis of FAD8-YFP over-expressing lines confirmed the chloroplast envelope localization of FAD8. Co-localization experiments suggested that FAD8 and FAD7 might be located in close vicinity in the envelope membrane. FAD8-YFP over-expressing lines showed a specific increase in 18:3 fatty acids at 22 °C. Together, these results indicate that the function of both plastidial ω-3 desaturases is coordinated in a non-redundant manner.