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Molecular cloning and functional analysis of the drought tolerance gene MsHSP70 from alfalfa (Medicago sativa L.)

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Abstract

Heat shock proteins (HSPs) are a ubiquitously expressed class of protective proteins that play a key role in plant response to stressful conditions. This study aimed to characterize and investigate the function of an HSP gene in alfalfa (Medicago sativa). MsHSP70, which contains a 2028-bp open reading frame, was identified through homology cloning. MsHSP70 shares high sequence identity (94.47%) with HSP70 from Medicago truncatula. Expression analysis of MsHSP70 in alfalfa organs revealed a relatively higher expression level in aerial organs such as flowers, stems and leaves than in roots. MsHSP70 was induced by heat shock, abscisic acid (ABA) and hydrogen peroxide. Transgenic Arabidopsis seedlings overexpressing MsHSP70 were hyposensitive to polyethylene glycol (PEG) and ABA treatments, suggesting that exogenous expression of MsHSP70 enhanced Arabidopsis tolerance to these stresses. Examination of physiological indexes related to drought and ABA stress demonstrated that in comparison with non-transgenic plants, T3 transgenic Arabidopsis plants had an increased proline content, higher superoxide dismutase (SOD) activity, and decreased malondialdehyde (MDA) content. Furthermore, higher relative water content (RWC) was detected in transgenic plants compared with non-transgenic plants under drought stress. These findings clearly indicate that molecular manipulation of MsHSP70 in plants can have substantial effects on stress tolerance.

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Abbreviations

HSP:

Heat shock protein

ABA:

Abscisic acid

SOD:

Superoxide dismutase

MDA:

Malondialdehyde

RWC:

Relative water content

ORF:

Open reading frame

kDa:

Kilodalton

GFP:

Green fluorescent protein

PEG:

Polyethylene glycol

qPCR:

Quantitative real-time PCR

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Acknowledgements

This work was supported by the National Program on Key Basic Research Project (Grant Number: 2014CB138703-2), the China Agriculture Research System (Grant Number: CARS-35-04) and The Agricultural Science and Technology Innovation Program (Grant Number: ASTIP-IAS14).

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Correspondence to Junmei Kang or Yan Sun.

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The authors declare that they have no conflicts of interest.

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Li, Z., Long, R., Zhang, T. et al. Molecular cloning and functional analysis of the drought tolerance gene MsHSP70 from alfalfa (Medicago sativa L.). J Plant Res 130, 387–396 (2017). https://doi.org/10.1007/s10265-017-0905-9

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  • DOI: https://doi.org/10.1007/s10265-017-0905-9

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