Involvement of reactive oxygen species and Ca2+ in the differential responses to low-boron in rapeseed genotypes
Background and aims
Boron (B) deficiency significantly inhibits plant growth and development. Oilseed rape (Brassica napus L.) is highly susceptible to B deficiency. Reactive oxygen species (ROS) and Ca2+ play pivotal roles in plant responses to environmental stresses. We aim to identify the differential Ca2+ fluxes and ROS bursts of a B-efficient genotype ‘QY10’ and a B-inefficient genotype ‘W10’ to B deficiency, and establish a signalling pathway involving Ca2+ and ROS implicated in the low-B-induced cell death.
Under both plant and suspension cell systems, the ROS production was investigated histochemically, cytochemically and biochemically; K+ and Ca2+ effluxes were assayed using the Non-invasive Micro-test Technology (NMT); the expression of ROS-producing genes and the activity assays of antioxidant enzymes were tested, and the ROS scavengers and Ca2+ channel inhibitors were used to characterize the roles of ROS and Ca2+ in response to low-B, respectively.
The cell death was mainly responsible for rapeseed growth inhibition under B deficiency. Low-B induced O2 − accumulation, whose distribution was similar to the cell death regions in the plant roots. The increase in O2 − production was much stronger in ‘W10’ than in ‘QY10’. The change trend of H2O2 was similar to that of O2 −, whereas less significant. The enhancement of lipid peroxidation, ion leakage and K+ efflux indicated that low-B caused cell death through the induction of oxidative damages, particularly in ‘W10’. Pretreatment with O2 − scavenger increased the cell viabilities. Low-B induced Ca2+ influx, which worked upstream of ROS. It was not the antioxidant enzymes but the ROS-generating enzymes that determined the differential oxidative damages in rapeseed genotypes.
Low-B induced Ca2+ influx, which then stimulated the ROS burst and eventually caused cell death. The present study enriches our understanding of the involvement of ROS and Ca2+ in the differential responses to B deficiency in rapeseed genotypes.
KeywordsBoron deficiency Brassica napus Ca2+ Genotypes Reactive oxygen species
Non-invasive Micro-test Technology
Programmed cell death
Quantitative real-time PCR
Respiratory Burst Oxidase Homolog
Reactive oxygen species
This work was financially supported by the National Natural Science Foundation of China (Grant NO. 31372129, 31572185) and the National Key Research and Development Program of China (Grant NO. 2016YFD0100700).
- Ahmad W, Niaz A, Zia MH, Saifullah, Malhi SS (2012) Boron deficiency in soils and crops: a review. INTECH Open Access PublisherGoogle Scholar
- Choudhury FK, Rivero RM, Blumwald E, Mittler R (2016) Reactive oxygen species, abiotic stress and stress combination. Plant J. doi: 10.1111/tpj.13299
- Diana G (2006) Boron in the soil, from deficit to toxicity. Informatore Agrario 62:54–58Google Scholar
- FAO (Food and Agriculture Organization). 2010. Food Balance Sheets. Available online: (http://faostat.fao.org/default.aspx), accessed July 30, 2010.
- Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil, 2nd edn. College of Agriculture, University of CaliforniaGoogle Scholar
- Hua YP, Zhang DD, Zhou T, He ML, Ding GD, Shi L, Xu FS (2016a) Transcriptomics-assisted quantitative trait locus fine mapping for the rapid identification of a nodulin 26-like intrinsic protein gene regulating boron efficiency in allotetraploid rapeseed. Plant Cell Environ 39:1601–1618CrossRefPubMedGoogle Scholar
- Lee B, Zhu JK (2010) Phenotypic analysis of Arabidopsis mutants: electrolyte leakage after freezing stress. Cold Spring Harb Protoc. doi: 10.1101/pdb.prot4970
- Levitt J (1972) Responses of plants to environmental stresses. Academic Press, New YorkGoogle Scholar
- Lherminier J, Elmayan T, Fromentin J, Elaraqui KT, Vesa S, Morel J, Verrier JL, Cailleteau B, Blein JP, Simon-Plas F (2009) NADPH oxidase-mediated reactive oxygen species production: subcellular localization and reassessment of its role in plant defense. Mol Plant-Microbe Interact 22:868–881CrossRefPubMedGoogle Scholar
- Miller G, Schlauch K, Tam R, Cortes D, Torres MA, Shulaev V, Dangl JL, Mittler R (2009) The plant NADPH oxidase RBOHD mediates rapid systemic signaling in response to diverse stimuli. Sci Signal 2: ra45Google Scholar
- Sun C, Wu T, Zhai L, Li D, Zhang X, Xu X, Ma H, Wang Y, Han Z (2016) Reactive oxygen species function to mediate the Fe deficiency response in an Fe-efficient apple genotype: An early response mechanism for enhancing reactive oxygen production. Front Plant Sci 7:1726PubMedPubMedCentralGoogle Scholar
- Wang YH, Shi L, Cao XY, Xu FS (2007) Studies on plant boron nutrition and boron fertilization in China. In Xu, FS, Goldbach HE, Brown PH, Bell, RW, Fujiwara T, Hunt CD, et al. (2007) Advances in plant and animal boron nutrition. Netherlands: Springer Press. pp. 93–101Google Scholar