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Overexpression of MpCYS2, a phytocystatin gene from Malus prunifolia (Willd.) Borkh., confers drought tolerance and protects against oxidative stress in Arabidopsis

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Abstract

Cystatins, or phytocystatins, are plant-specific inhibitors of cysteine proteinases that help regulate endogenous processes and protect plants against heat, salinity, cold, water deficits, chilling, and abscisic acid treatment. MpCYS2, a cystatin gene from Malus prunifolia, localized to the nucleus, cytoplasm, and plasma membrane in onion epidermal cells. Controlled by the 35S promoter, its ectopic expression in transgenic Arabidopsis lines caused accelerated seed germination and greater seedling growth when plants were exposed to osmotic or oxidative stress. Expression by this gene was also associated with enhanced drought tolerance in those transgenics. This positive response was manifested by changes measured in electrolyte leakage, the chlorophyll concentration, and malondialdehyde accumulations. Production of reactive oxygen species (ROS) was appreciably decreased in the dehydration-treated transgenic lines. This gene also influenced root hair development under osmotic-stress conditions. Our findings indicate that MpCYS2 affects the growth and tolerance of drought-stressed Arabidopsis plants possibly because of its influence on ROS accumulations and root hair formation.

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Abbreviations

ABA:

Abscisic acid

DAB:

Diaminobenzidine

EL:

Electrolyte leakage

H2O2 :

Hydrogen peroxide

Km:

Kanamycin

MDA:

Malondialdehyde

MS:

Murashige and Skoog

MV:

Methyl viologen

NBT:

Nitro blue tetrazolium

O2 − :

Superoxide anion

ORF:

Open reading frame

PhyCys:

Phytocystatins

qRT-PCR:

Quantitative real-time PCR

ROS:

Reactive oxygen species

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program) (2011AA100201) and by the earmarked fund for the China Agriculture Research System (CARS-28).

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Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Yanxiao Tan, Mingjun Li & Fengwang Ma

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  1. Yanxiao Tan
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  2. Mingjun Li
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Correspondence to Fengwang Ma.

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Supplementary Table S1 Primers used in this study (DOC 27 kb)

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Tan, Y., Li, M. & Ma, F. Overexpression of MpCYS2, a phytocystatin gene from Malus prunifolia (Willd.) Borkh., confers drought tolerance and protects against oxidative stress in Arabidopsis . Plant Cell Tiss Organ Cult 123, 15–27 (2015). https://doi.org/10.1007/s11240-015-0809-0

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