Genotypic difference in response to copper stress in upland cotton as revealed by physiological and molecular expression analyses

Background: Cotton is a non-edible fiber crop with considerable potential for the remediation of copper-polluted soil. However, the Cu toxicity tolerance mechanism in cotton remains largely obscure. To address the issue, we first identified two cotton lines contrasting in response to Cu toxicity by examining 12 morphological and physiological attributes of 43 origin scattered cotton genotypes under Cu excess. Then both lines were subjected to a comprehensive comparative study, aiming to unravel the cotton Cu tolerance mechanism through integrated morphological, physio-biochemical, Cu uptake and distribution, and related molecular expression analyses.

Results: Based on the phenotypic values and corresponding tolerance indexes of 12 parameters, A2304 and A1415 were identified as Cu-tolerant and -sensitive, respectively. Compared to A1415, A2304 exhibited significantly higher antioxidant enzyme activities and non-enzymatic antioxidant levels, producing fewer amounts of reactive oxygen species and a lower level of malonyldialdehyde. On Cu excess, A2304 accumulated lower concentrations of Cu ions in various plant parts and subcellular components, and fewer Cu ions were presented in active chemical forms. However, the total Cu uptake amount per plant did not differ between both lines due to larger plant biomass with A2304. In contrast to A1415, Cu stress activated or increased the expressions of Cu homeostasis regulator (GhSPL7) and genes responsible for Cu delivery (GhCCS, GhCOX17), chelation (GhMT2), and compartmentation into vacuoles (GhHMA5), while inactivating or decreasing the expressions of genes accounting for Cu uptake (GhCOPT1) and Cu exporting from vacuoles (GhCOPT5) in the root cell with A2304. Additionally, A2304 may impede the root cell wall from binding Cu ions by enhancing the pectin methylesterification degree by up-regulating GhPMEI3 and GhPMEI9 encoding pectin methylesterase inhibitor and stabilizing the cell wall organization by down-regulating GhPLY8 and GhPLY20 encoding pectate lyases.

Conclusions: To cope with Cu toxicity, the Cu-tolerant genotype activates its antioxidative defense system, immobilizing chemically active Cu ions, and lowering the Cu uptake, bioavailability and immigration within cells by regulating the expressions of genes related to Cu uptake, transport, delivery and cell wall metabolism. This comprehensive comparison study provides insights into breeding Cu-tolerant cotton cultivars that can be utilized for the phytoremediation of Cu-contaminated soils.