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Cloning and characterization of a heme oxygenase-2 gene from rice (Oryza sativa L.), and its expression analysis in response to some abiotic stresses

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Abstract

Heme oxygenase (HO; EC 1.14.99.3) is an important enzyme that yields biliverdin IXα (BV), carbon monoxide and iron. At least two kinds of HO subfamilies exist in plants. Our previous report revealed that rice (Oryza sativa L.) HO1 (SE5, also named as OsHO1), a major subfamily of HO, is an oxidative stress–response protein, especially upon paraquat exposure. However, whether there exists rice HO2, another subfamily of HO, is still unknown. In the present study, a rice HO2 gene (named as OsHO2) was cloned and characterized. Its genomic sequence consists of four exons and three introns, and encodes a 36.5 kDa protein precursor with a 4.9 kDa N-terminal transit leader peptide. Further results showed that OsHO2 has a conserved HO signature sequence and shares a high amino acid sequence similarity with other identified plant HO2s. The recombinant mature OsHO2 (mOsHO2) protein expressed in Escherichia coli did not exhibit HO activity, which was in contrast with that of mOsHO1. The results of subcellular localization of OsHO2 demonstrated that it was most likely localized in the chloroplasts. Real-time RT-PCR experiment revealed that although OsHO2 mRNA is a much less abundant than that of OsHO1, both of them were expressed in all tested tissues. Importantly, OsHO2 transcripts could be differentially induced by hemin (a substrate of HO), paraquat (in particular), and NaCl treatments. Together, the results suggested that OsHO2 might act downstream in the signal transduction pathways following abiotic stresses in rice.

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Abbreviations

AMV:

Avian myeloblastosis virus

BV:

Biliverdin IXα

CO:

Carbon monoxide

CTAB:

Cetyltrimethylammonium bromide

EST:

Expressed sequence tags

HO:

Heme oxygenase

NO:

Nitric oxide

PBS:

Phosphate-buffered saline

PQ:

Paraquat

ROS:

Reactive oxygen species

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Acknowledgments

We greatly appreciate Dr. Judy C Lee for her kindly help in the improvement of English writing. This work was supported by the projects (30960182 and 31000700) from the National Natural Science Foundation of China, the “948” project (2011-G2B(3)) from the Ministry of Agriculture of China, the “863” project (2012AA101101) from China’s Ministry of Science & Technology, and the union project from Bill & Melinda Gates Foundation and China’s Ministry of Science & Technology.

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Authors and Affiliations

  1. Laboratory Center of Life Sciences, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China

    Lijuan Wang, Fei Ma & Wenbiao Shen

  2. Institute of Botany, Jiangsu Province and the Chinese Academy of Sciences, Jiangsu Province Key Laboratory for Plant Ex-situ Conservation, Nanjing, 210014, China

    Sheng Xu & Ren Wang

  3. Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, 12 South Zhong-Guan-Cun St., Beijing, 100081, China

    Tianqing Zheng

  4. College of Horticulture and Garden, Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources, Hainan University, Haikou, 570228, China

    Huiping Chen

  5. College of Horticulture and Landscape Architecture, Hainan University, Haikou, 570228, China

    Huiping Chen

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  1. Lijuan Wang
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  3. Sheng Xu
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Correspondence to Wenbiao Shen.

Additional information

Communicated by T. Moriguchi.

L. Wang and F. Ma contributed equally to the work.

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Wang, L., Ma, F., Xu, S. et al. Cloning and characterization of a heme oxygenase-2 gene from rice (Oryza sativa L.), and its expression analysis in response to some abiotic stresses. Acta Physiol Plant 36, 893–902 (2014). https://doi.org/10.1007/s11738-013-1468-6

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  • DOI: https://doi.org/10.1007/s11738-013-1468-6

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