Stomata regulation by tissue-specific expression of the Citrus sinensis MYB61 transcription factor improves water-use efficiency in Arabidopsis.

Water-use efficiency (WUE) is a quantitative measurement of biomass produced per volume of water transpired by a plant. WUE is an important physiological trait for drought response to mitigate the water deficiency. In this work, a cisgenic construction from Citrus sinensis was developed and its function in the improvement of WUE was evaluated in Arabidopsis. Sequences of the CsMYB61 coding region, a transcription factor implicated in the closure of stomata, together with a putative stomata-specific promoter from CsMYB15, were identified and cloned. The protein encoded in the CsMYB61 locus harbors domains and motifs characteristic of MYB61 proteins. In addition, a 1.2 kb promoter region of the gene CsMYB15 (pCsMYB15) containing regulatory elements for expression in guard cells and in response to Abscisic Acid (ABA) and light was isolated. In Arabidopsis, pCsMYB15 directs the expression of the reporter gene GUS in stomata in the presence of light. In addition, transgenic lines expressing the CsMYB61 coding region under transcriptional control of pCsMYB15 have a normal phenotype under in vitro and greenhouse conditions. These transgenic lines exhibited a smaller opening of the stomata pore, lower stomatal conductance and respiration rate, enhanced sensitivity to exogenous ABA, and high drought stress tolerance. Our results indicate that stomata-specific expression of CsMYB61 enhances water use efficiency under drought conditions in Arabidospis.

Molecular characterization of a Trithorax-group homologue gene from Pinus radiata.

In eukaryotes, trithorax group proteins play critical roles in the regulation of transcription, cell proliferation, differentiation and development. In this work we report the molecular cloning and characterization of SEPR11, a cDNA from the conifer Monterrey pine (Pinus radiata) encoding a polypeptide homologue of a trithorax group member described in animals and yeast. A full-length clone was isolated from RNA prepared from somatic embryos and contained a 1,239 bp ORF encoding 412 amino acids. Characterization of the isolated sequence revealed that it contains a SPRY domain in the C-terminal region. A comparison of the pine sequence with homologous proteins from plants, animals and yeast revealed that SEPR11 is phylogenetically related to the trithorax group members and not a SPRY-domain containing protein. RT-PCR analyses of transcript abundance in pine tissues demonstrated that SEPR11 is particularly abundant in embryos, suggesting that this gene could be involved during embryo development. The spatial localization of SEPR11 transcripts revealed that gene expression was restricted to the vascular bundle and apical and radicular meristems, suggesting a possible function of this gene in meristem control and vascular bundle development. This work is the first report of the presence of a trithorax group homologue gene in gymnosperm.