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Abiotic stress induces change in Cinnamoyl CoA Reductase (CCR) protein abundance and lignin deposition in developing seedlings of Leucaena leucocephala

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Abstract

Aboitic stress such as drought and salinity are class of major threats, which plants undergo through their lifetime. Lignin deposition is one of the responses to such abiotic stresses. The gene encoding Cinnamoyl CoA Reductase (CCR) is a key gene for lignin biosynthesis, which has been shown to be over-expressed under stress conditions. In the present study, developing seedlings of Leucaena leucocephala (Vernacular name: Subabul, White popinac) were treated with 1 % mannitol and 200 mM NaCl to mimic drought and salinity stress conditions, respectively. Enzyme linked immunosorbant assay (ELISA) based expression pattern of CCR protein was monitored coupled with Phlorogucinol/HCl activity staining of lignin in transverse sections of developing L. leucocephala seedlings under stress. Our result suggests a differential lignification pattern in developing root and stem under stress conditions. Increase in lignification was observed in mannitol treated stems and corresponding CCR protein accumulation was also higher than control and salt stress treated samples. On the contrary CCR protein was lower in NaCl treated stems and corresponding lignin deposition was also low. Developing root tissue showed a high level of CCR content and lignin deposition than stem samples under all conditions tested. Overall result suggested that lignin accumulation was not affected much in case of developing root however developing stems were significantly affected under drought and salinity stress condition.

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Abbreviations

BCIP/NBT:

5-bromo-4-chloro-3-indolyl-phosphate/ nitro blue tetrazolium

C3H:

Cinnamate 3 hydroxylase

C4H:

Cinnamate 4 hydroxylase

COMT:

Caffeate O-methyl transferase

CCoAOMT:

Caffeoyl CoA O-methyl transferase

DTT:

Dithiothreitol

EDTA:

Ethylenediaminetetraacetic acid

ELISA:

Enzyme-linked immunosorbent assay

F5H:

Ferulate 5-hydroxylase

FTIR:

Fourier transform infrared spectroscopy

HIS:

6 Histidine-tag

HCT:

Hydroxy cinnamoyl transferase

KBr:

Potassium bromide

Ni-NTA:

Ni-nitrilotriacetic acid

PEG:

Poly ethylene glycol

PMSF:

Phenylmethylsulfonyl fluoride

PVPP:

Polyvinyl polypyrrolidone

RT:

Room temperature

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Acknowledgments

This work was supported by grants from Council of Scientific and Industrial Research-NMITLI, India. SS acknowledge the financial assitance (fellowship) provided by CSIR-UGC. SKG is thankful to CSIR for providing financial assistance in form of fellowship. RKV and YAA are thankful to CSIR-NMITLI for their fellowship grants. The authors wish to thank Ms. T. Kamat for proof reading of the manuscript.

Conflicts of interest

The authors declare that they have no conflicts of interest.

Authors’ contributions

SS and BMK conceived, designed the experiments, and wrote the manuscript. SS, RKV, SKG and YAA performed the experiments. All authors read and approved the final manuscript.

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  1. Sameer Srivastava

    Present address: Department of Biotechnology, Motilal Nehru National Institute of Technology, Allahabad, 211002, India

Authors and Affiliations

  1. Plant Tissue Culture Division, National Chemical Laboratory, Dr. Homi Bhabha Road, PAshan, Pune, Maharashtra, 411008, India

    Sameer Srivastava, Rishi K. Vishwakarma, Yasir Ali Arafat, Sushim K. Gupta & Bashir M. Khan

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Correspondence to Bashir M. Khan.

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Srivastava, S., Vishwakarma, R.K., Arafat, Y.A. et al. Abiotic stress induces change in Cinnamoyl CoA Reductase (CCR) protein abundance and lignin deposition in developing seedlings of Leucaena leucocephala . Physiol Mol Biol Plants 21, 197–205 (2015). https://doi.org/10.1007/s12298-015-0289-z

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