Lara Lombardi

Physiological, Biochemical, and Molecular Effects of In Vitro Induced Iron Deficiency in Peach Rootstock Mr.S 2/5

Abstract On calcareous soils, carbonate induced iron deficiency can have heavy effects on growth and development of several fruit crops. Leaf chlorosis, biochemical, and ultrastructural alterations are the first symptoms causing severe damages to yield and productivity. In this work, some physiological, biochemical, and molecular aspects of iron deficiency stress induced on the peach rootstock Mr.S 2/5 in vitro were studied. The aim was to demonstrate if in vitro culture can be used as a valid and fast method to evoke iron deficiency symptoms and to study plant responses to this kind of abiotic stress. Two different treatments were carried out and compared to a control (MS medium): plantlets grown on MS medium completely free of iron and on MS medium containing 1 mM potassium bicarbonate. After 10 and 20 days from the beginning of the treatments fresh and dry weight, chlorophyll, and carotenoids content were measured. Superoxide dismutase and catalase total activity was measured, and gene expression was analyzed by using etherologous probes (Sod1, Sod3, Sod4, and Cat1) obtained from maize. In the absence of iron or in the presence of bicarbonate Mr.S 2/5 plantlets showed a significant decrease in growth, as quantified by fresh and dry weight. The plantlets showed a severe chlorosis due to a reduction in chlorophyll and carotenoids concentration. Iron deficiency stress caused a reduction of the activity of catalase and superoxide dismutase, while inducing an increase in gene expression.

RNA sequencing of Populus x canadensis roots identifies key molecular mechanisms underlying physiological adaption to excess zinc

Populus x canadensis clone I-214 exhibits a general indicator phenotype in response to excess Zn, and a higher metal uptake in roots than in shoots with a reduced translocation to aerial parts under hydroponic conditions. This physiological adaptation seems mainly regulated by roots, although the molecular mechanisms that underlie these processes are still poorly understood. Here, differential expression analysis using RNA-sequencing technology was used to identify the molecular mechanisms involved in the response to excess Zn in root. In order to maximize specificity of detection of differentially expressed (DE) genes, we consider the intersection of genes identified by three distinct statistical approaches (61 up- and 19 down-regulated) and validate them by RT-qPCR, yielding an agreement of 93% between the two experimental techniques. Gene Ontology (GO) terms related to oxidation-reduction processes, transport and cellular iron ion homeostasis were enriched among DE genes, highlighting the importance of metal homeostasis in adaptation to excess Zn by P. x canadensis clone I-214. We identified the up-regulation of two Populus metal transporters (ZIP2 and NRAMP1) probably involved in metal uptake, and the down-regulation of a NAS4 gene involved in metal translocation. We identified also four Fe-homeostasis transcription factors (two bHLH38 genes, FIT and BTS) that were differentially expressed, probably for reducing Zn-induced Fe-deficiency. In particular, we suggest that the down-regulation of FIT transcription factor could be a mechanism to cope with Zn-induced Fe-deficiency in Populus. These results provide insight into the molecular mechanisms involved in adaption to excess Zn in Populus spp., but could also constitute a starting point for the identification and characterization of molecular markers or biotechnological targets for possible improvement of phytoremediation performances of poplar trees.