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Molecular characterization and primary functional analysis of PeMPEC, a magnesium-protoporphyrin IX monomethyl ester cyclase gene of bamboo (Phyllostachys edulis)

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Abstract

Key message

A homologous gene of MPEC from Phyllostachys edulis was isolated and characterized. Its overexpression in Arabidopsis thaliana increased chlorophyll concentration and photosynthesis efficiency, indicating it is involved in chlorophyll biosynthesis.

Abstract

Magnesium-protoporphyrin IX monomethyl ester cyclase (MPEC) is an essential enzyme in the biosynthesis of chlorophyll, which plays an important role in photosynthesis. However, limited information is available on the roles of MPEC gene in bamboo. A homologous gene, PeMPEC was identified from Phyllostachys edulis, which comprised 1474 bp and contained an open reading frame encoding 415 amino acids. PeMPEC was transcribed abundantly in leaf blade where photosynthesis predominantly occurs, which agreed with the protein accumulation pattern confirmed by Western blotting. The PeMPEC transcription was promoted by continuous darkness for 24 h, and was suppressed by increasing light intensity (100–1500 µmo1 m−2 s−1) and high temperature (42 °C). However, transcription was induced within 0.5 h and thereafter declined with prolonged treatment (up to 12 h) under low temperature (4 °C). Although PeMPEC expressed weakly in etiolated leaves, transcript levels increased gradually with subsequent light treatment (200 µmol m−2 s−1). Overexpression of PeMPEC in Arabidopsis thaliana resulted in increased chlorophyll concentration and photosynthesis efficiency in sense transgenic plants compared with a reduction in antisense transgenic plants. These changes were consistent with the transcript levels of PeMPEC. These results indicated that PeMPEC might be involved in chlorophyll biosynthesis and play important roles in maintaining the stability of photosystems, and provide a basis for the study of chlorophyll biosynthesis and dissection of photosynthesis in bamboo.

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Abbreviations

cDNA:

Complementary DNA

F o :

Minimal fluorescence

F m :

Maximal fluorescence after darkness adaptation

F m′:

Maximum fluorescence after light adaptation

F s :

Steady-state fluorescence after light adaptation

F v :

Variable fluorescence

F v/F m :

Maximal quantum yield of PSII

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

kDa:

Kilodaltons

Mg:

Magnesium

MPEC:

Mg-protoporphyrin IX monomethyl ester cyclase

ORF:

Open reading frame

P m :

Maximum photo-oxidizable P700

PSI:

Photosystem I

PSII:

Photosystem II

qPCR:

Real-time quantitative PCR

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

Y(II):

Maximal actual quantum yield of PSII

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Acknowledgments

This work received financial support from the Special Fund for Forest Scientific Research in the Public Welfare from State Forestry Administration of China (No. 201504106) and the National Science Foundation of China (No. 31370588 and No. 31400557).

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    Authors and Affiliations

    1. Key Laboratory on the Science and Technology of Bamboo and Rattan, International Center for Bamboo and Rattan, Beijing, 100102, People’s Republic of China

      Li Yang, Yongfeng Lou, Zhenhua Peng, Hansheng Zhao, Huayu Sun & Zhimin Gao

    2. Institute of Gene Science for Bamboo and Rattan Resources, Beijing, 100102, People’s Republic of China

      Li Yang, Yongfeng Lou, Hansheng Zhao, Huayu Sun & Zhimin Gao

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    1. Li Yang
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    Correspondence to Zhimin Gao.

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    Communicated by B. Li.

    L. Yang and Y. Lou contributed equally to this work.

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    Yang, L., Lou, Y., Peng, Z. et al. Molecular characterization and primary functional analysis of PeMPEC, a magnesium-protoporphyrin IX monomethyl ester cyclase gene of bamboo (Phyllostachys edulis). Plant Cell Rep 34, 2001–2011 (2015). https://doi.org/10.1007/s00299-015-1846-1

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