title: Regulation of copper homeostasis and biotic interactions by microRNA 398b in common bean.
abstract: MicroRNAs are recognized as important post-transcriptional regulators in plants. Information about the roles of miRNAs in common bean (Phaseolus vulgaris L.), an agronomically important legume, is yet scant. The objective of this work was to functionally characterize the conserved miRNA: miR398b and its target Cu/Zn Superoxide Dismutase 1 (CSD1) in common bean. We experimentally validated a novel miR398 target: the stress up-regulated Nodulin 19 (Nod19). Expression analysis of miR398b and target genes -CSD1 and Nod19- in bean roots, nodules and leaves, indicated their role in copper (Cu) homeostasis. In bean plants under Cu toxicity miR398b was decreased and Nod19 and CSD1, that participates in reactive oxygen species (ROS) detoxification, were up-regulated. The opposite regulation was observed in Cu deficient bean plants; lower levels of CSD1 would allow Cu delivery to essential Cu-containing proteins. Composite common bean plants with transgenic roots over-expressing miR398 showed ca. 20-fold higher mature miR398b and almost negligible target transcript levels as well as increased anthocyanin content and expression of Cu-stress responsive genes, when subjected to Cu deficiency. The down-regulation of miR398b with the consequent up-regulation of its targets was observed in common bean roots during the oxidative burst resulting from short-time exposure to high Cu. A similar response occurred at early stage of bean roots inoculated with Rhizobium tropici, where an increase in ROS was observed. In addition, the miR398b down-regulation and an increase in CSD1 and Nod19 were observed in bean leaves challenged with Sclerotinia scleortiorum fungal pathogen. Transient over-expression of miR398b in Nicotiana benthamiana leaves infected with S. sclerotiorum resulted in enhanced fungal lesions. We conclude that the miR398b-mediated up-regulation of CSD and Nod19 is relevant for common bean plants to cope with oxidative stress generated in abiotic and biotic stresses.
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: Loreto
authors.initials: L
authors.lastname: Naya
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: Sujay
authors.initials: S
authors.lastname: Paul
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México ; Laboratorio de Bioquímica, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México. Tlalnepantla, Estado de México. México.
authors.forename: Oswaldo
authors.initials: O
authors.lastname: Valdés-López
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: Ana B
authors.initials: AB
authors.lastname: Mendoza-Soto
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: Bárbara
authors.initials: B
authors.lastname: Nova-Franco
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: Guadalupe
authors.initials: G
authors.lastname: Sosa-Valencia
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: José L
authors.initials: JL
authors.lastname: Reyes
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos. México.
authors.forename: Georgina
authors.initials: G
authors.lastname: Hernández
mesh_terms: D003300:Copper / Q000378:metabolism*; D007887:Fabaceae / Q000502:physiology*; D018506:Gene Expression Regulation, Plant; D006706:Homeostasis; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D010641:Phenotype; D017382:Reactive Oxygen Species
chemical_list: D035683:MicroRNAs; D017382:Reactive Oxygen Species; D003300:Copper