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Plant and Soil

, Volume 419, Issue 1–2, pp 219–236 | Cite as

Involvement of reactive oxygen species and Ca2+ in the differential responses to low-boron in rapeseed genotypes

  • Ting Zhou
  • Yingpeng Hua
  • Fangsen Xu
Regular Article

Abstract

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.

Methods

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.

Results

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.

Conclusions

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.

Keywords

Boron deficiency Brassica napus Ca2+ Genotypes Reactive oxygen species 

Abbreviations

ASC

Ascorbate

B

Boron

CAT

Catalase

DAB

3, 3′-diaminobenzidine

DCF-DA

2′,7′-dichlorofluorescein diacetate

DHE

Dihydroethidium

FDA

Fluorescein diacetate

H2O2

Hydrogen peroxide

MDA

Malondialdehyde

NBT

Nitroblue tetrazolium

NMT

Non-invasive Micro-test Technology

O2

Superoxide radical

PCD

Programmed cell death

PI

Propidium iodide

PM

Plasma membrane

POD

Peroxidase

qRT-PCR

Quantitative real-time PCR

RBOH

Respiratory Burst Oxidase Homolog

ROS

Reactive oxygen species

RGII

Rhamnogalacturonan II

SOD

Superoxide dismutase

TMP

Tetramethyl piperidinooxy

Notes

Acknowledgements

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).

Supplementary material

11104_2017_3337_MOESM1_ESM.docx (586 kb)
ESM 1 (DOCX 586 kb)

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Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  1. 1.National Key Laboratory of Crop Genetic Improvement and Microelement Research CentreHuazhong Agricultural UniversityWuhanPeople’s Republic of China

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