title: Photosynthesis light-independent reactions are sensitive biomarkers to monitor lead phytotoxicity in a Pb-tolerant Pisum sativum cultivar.
abstract: Lead (Pb) environmental contamination remains prevalent. Pisum sativum L. plants have been used in ecotoxicological studies, but some cultivars showed to tolerate and accumulate some levels of Pb, opening new perspectives to their use in phytoremediation approaches. However, the putative use of pea plants in phytoremediation requires reliable toxicity endpoints. Here, we evaluated the sensitivity of a large number of photosynthesis-related biomarkers in Pb-exposed pea plants. Plants (cv. "Corne de Bélier") were exposed to Pb concentrations up to 1,000 mg kg(-1) soil during 28 days. The photosynthetic potential biomarkers that were analyzed included pigments, chlorophyll (Chl) a fluorescence, gas exchange, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity, and carbohydrates. Flow cytometry (FCM) was also used to assess the morpho-functional status of chloroplasts. Finally, Pb-induced nutrient disorders were also evaluated. Net CO2 assimilation rate (A) and RuBisCO activity decreased strongly in Pb-exposed plants. Plant dry mass (DM) accumulation, however, was only reduced in the higher Pb concentrations tested (500 and 1,000 mg kg(-1) soil). Pigment contents increased solely in plants exposed to the largest Pb concentration, and in addition, the parameters related to the light-dependent reactions of photosynthesis, Fv/Fm and ΦPSII, were not affected by Pb exposure. In contrast to this, carbohydrates showed an overall tendency to increase in Pb-exposed plants. The morphological status of chloroplasts was affected by Pb exposure, with a general trend of volume decrease and granularity increase. These results point the endpoints related to the light-independent reactions of photosynthesis as more sensitive predictors of Pb-toxicity than the light-dependent reactions ones. Among the endpoints related to the light-independent photosynthesis reactions, RuBisCO activity and A were found to be the most sensitive. We discuss here the advantages of using these parameters as biomarkers for Pb toxicity in plants. Finally, we report that, despite showing physiological disorders, these cultivar plants survived and accumulated high doses of Pb, and their use in environmental/decontamination studies is open to debate.
authors.affiliation: Department of Biology, Laboratory of Biotechnology and Cytometry, Campus Universitário de Santiago, University of Aveiro, CESAM, 3810, Aveiro, Portugal.
authors.forename: Eleazar
authors.initials: E
authors.lastname: Rodriguez
authors.forename: Maria
authors.initials: M
authors.lastname: da Conceição Santos
authors.forename: Raquel
authors.initials: R
authors.lastname: Azevedo
authors.forename: Carlos
authors.initials: C
authors.lastname: Correia
authors.forename: José
authors.initials: J
authors.lastname: Moutinho-Pereira
authors.forename: José Miguel Pimenta
authors.initials: JM
authors.lastname: Ferreira de Oliveira
authors.forename: Maria Celeste
authors.initials: MC
authors.lastname: Dias
mesh_terms: D001673:Biodegradation, Environmental; D015415:Biomarkers / Q000378:metabolism; D050260:Carbohydrate Metabolism / Q000187:drug effects; D002734:Chlorophyll / Q000378:metabolism; D000077194:Chlorophyll A; D002736:Chloroplasts / Q000187:drug effects / Q000378:metabolism; D004784:Environmental Monitoring; D005434:Flow Cytometry; D005453:Fluorescence; D007854:Lead / Q000378:metabolism / Q000633:toxicity*; D008027:Light; D018532:Pisum sativum / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D010788:Photosynthesis / Q000187:drug effects*; D010944:Plants / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism; D012987:Soil; D018675:Toxicity Tests
chemical_list: D015415:Biomarkers; D012987:Soil; D002734:Chlorophyll; D007854:Lead; D012273:Ribulose-Bisphosphate Carboxylase; D000077194:Chlorophyll A
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