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Characterization of the Copper Transporters from Lotus spp. and Their Involvement under Flooding Conditions.

ABSTRACT: Forage legumes are an important livestock nutritional resource, which includes essential metals, such as copper. Particularly, the high prevalence of hypocuprosis causes important economic losses to Argentinian cattle agrosystems. Copper deficiency in cattle is partially due to its low content in forage produced by natural grassland, and is exacerbated by flooding conditions. Previous results indicated that incorporation of Lotus spp. into natural grassland increases forage nutritional quality, including higher copper levels. However, the biological processes and molecular mechanisms involved in copper uptake by Lotus spp. remain poorly understood. Here, we identify four genes that encode putative members of the Lotus copper transporter family, denoted COPT in higher plants. A heterologous functional complementation assay of the Saccharomyces cerevisiae ctr1∆ctr3∆ strain, which lacks the corresponding yeast copper transporters, with the putative Lotus COPT proteins shows a partial rescue of the yeast phenotypes in restrictive media. Under partial submergence conditions, the copper content of L. japonicus plants decreases and the expression of two Lotus COPT genes is induced. These results strongly suggest that the Lotus COPT proteins identified in this work function in copper uptake. In addition, the fact that environmental conditions affect the expression of certain COPT genes supports their involvement in adaptive mechanisms and envisages putative biotechnological strategies to improve cattle copper nutrition.

SUBMITTER: Escaray FJ

PROVIDER: S-EPMC6651048 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Purpose: The State of Rio Grande do Sul is the largest producer of peaches from Brazil. However, it still has low values of productivity when compared to other States. One of the problems associated to it this is the occurrence of drainage soils problems, which can suffer flooding situations potentially hampering the development and productivity such culture. For studies to assist in the selection of flood tolerant genotypes, it is essential to understand the physiological and molecular changes of the plants in situations of oxygen deprivation. Using Illumina Hiseq2500 we performed transcriptome analysis of leaves from ‘Capdeboscq’ (Prunus persica) and ‘Julior’ (Prunus insititia x Prunus domestica) rootstocks under flooding for 48 hours. Methods: The mRNA of Prunus spp. plants cv. Capdeboscq e Julior was generated using deep sequencing, in triplicate, using Illumina Hi-Seq 2500, for the following treatments:I) control: plants received irrigation daily until field capacity; and II) plants exposed to flood stress, maintaining a water level of approximately 3 cm above the ground. The sequence reads that passed quality filters were analyzed at the transcript level using this method: Mapping using STAR and identification of differentially expressed genes (DEGs) was performed with the edgeR (false discovery rates - FDRs of <0.05). RT–qPCR validation was performed using SYBR Green assays. Results: Flooding stress causes important high transcriptional changes in the ‘Capdeboscq’ compared to 'Julior' and this is mainly due to their sensitivity/tolerance levels. ‘Capdeboscq’ had photosynthesis as the most affected physiological process at the molecular level, showing a large number of down-regulated enriched GOs, even though it activated cellular signaling pathways under flooding. 'Julior' was more efficient in defense responses, which include the activation of flavonoid biosynthesis pathways. Conclusions: The analysis of two Prunus spp. rootstocks contrasting to the level of tolerance / sensitivity provide new insights into the process of plant flood stress tolerance.

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Characterization of the Copper Transporters from Lotus spp. and Their Involvement under Flooding Conditions.

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