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Differential Expression of Serine/Threonine Protein Phosphatase Type-2C Under Drought Stress in Maize

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Abstract

Differential display reverse transcription polymerase chain reaction was used to study the messenger RNA differential expression of a drought tolerant inbred line ‘81565’ under drought stress and well-watered control. Fragments of MD1, MD2, and MD3 were found to be differentially expressed under drought stress. While the expression of MD1 and MD2 was down-regulated, the expression of MD3 was up-regulated. Sequence analysis of the three fragments revealed that MD1 shared 97% nucleotide sequence identity with maize chloroplast matK, a gene encoding RNA maturase involved in group II intron splicing of RNA transcript; MD2 had 99% identity with PP2C, a gene encoding serine/threonine phosphorylase 2C in resurrection grass Sporobolus stapfianus; and MD3 exhibited 99% sequence identity with a rice gene encoding metacaspase, an arginine/lysine-specific cysteine protease. Based on MD2 fragment sequence, a full-length complementary DNA of 1,731 bp was isolated. An open reading frame of 1,167 bp was found and predicted to encode a protein of 388 amino acids, containing the catalytic domain and conserved residues of serine/threonine protein phosphatase 2C. This open reading frame was identified as a new member of the PP2C gene family in maize and designated as ZmPP2Ca. The differential expression of ZmPP2Ca and enzyme activity of PP2C were also investigated in three drought-tolerant and two sensitive maize inbred lines under drought stress and well-watered (control) conditions by real-time fluorescence quantitative PCR and a non-radioactive labeled method, respectively. Down-regulation of the ZmPP2Ca transcript was observed in all three drought tolerant lines, whereas transcript levels were up-regulated in the two sensitive lines under drought stress. In light of the importance of protein kinases in signal transduction pathways, it is proposed that ZmPP2Ca may play a role in the signal transduction of maize in response to drought stress.

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Abbreviations

ABA:

abscisic acid

C T :

cycle threshold

DDRT-PCR:

differential display reverse transcription PCR

FQ-PCR:

fluorescence quantitative PCR

ORF:

open reading frame

PEG-6000:

polyethylene glycol 6000

PP:

serine/threonine protein phosphatases

PP2C:

serine/threonine protein phosphatase type-2C

PTP:

protein tyrosine phosphatase

RDA:

representational difference analysis

RT-PCR:

revere transcription PCR

SMART:

simple modular architecture research tool

SSH:

suppression subtractive hybridization

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (30571172 and 30671309), Rockefeller Foundation (2004 FS 047), and Program for Changjiang Scholar and Innovative Research Team in University (PCSIRT, IRT0453). The authors are grateful to Dr. Li Li for prereviewing this manuscript.

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  1. Maize Research Institute/Key Laboratory of Crop Genetic Resources and Improvement, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China

    Fu-Hua Li, Feng-Ling Fu, Li-Na Sha, Liang He & Wan-Chen Li

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  1. Fu-Hua Li
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  2. Feng-Ling Fu
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  3. Li-Na Sha
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  4. Liang He
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  5. Wan-Chen Li
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Correspondence to Wan-Chen Li.

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Li, FH., Fu, FL., Sha, LN. et al. Differential Expression of Serine/Threonine Protein Phosphatase Type-2C Under Drought Stress in Maize. Plant Mol Biol Rep 27, 29–37 (2009). https://doi.org/10.1007/s11105-008-0051-6

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