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Identification of differentially expressed genes preferably related to drought response in pigeon pea (Cajanus cajan) inoculated by arbuscular mycorrhizae fungi (AMF)

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Abstract

Pigeon pea is an ideal crop for sustainable agriculture systems in Karst areas of southwest China, which frequently suffers from the formidable water deficit. Physiologically, arbuscular mycorrhizae (AM)-colonized pigeon pea (Cajanus cajan) demonstrated a further enhanced tolerance to drought stress. To elucidate the molecular mechanism underlying the elevated tolerance, suppression subtractive hybridization (SSH) were employed to dig up the differentially expressed genes using mixed cDNAs prepared from drought-stressed and unstressed pigeon pea seedlings inoculated by AM fungi (AMF) in the present work. Both forward and reverse SSH cDNA library were constructed and a total of 768 clones were obtained. Dot-blotting expression analysis identified that 142 clones were upregulated, and 49 were downregulated during water stress. After sequencing, 182 unique expressed sequence tags (ESTs) were obtained via blast analysis, among which 142 (78%) exhibited high homology to previously identified or putative proteins, however, 40 (22%) showed no homology in the database. The upregulated (102) and downregulated (40) ESTs with significant protein homology might be sorted into 16 and 12 functional categories respectively, which involved in a broad spectrum of biological pathways. Furthermore, semi-quantitative reverse transcription (RT)-PCR was carried out for the 35 differentially expressed genes whose putative functions implicated in abiotic stress tolerances in other species, and it was verified these differentially expressed genes highly involved in drought stress tolerance of AM-colonized pigeon pea.

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Abbreviations

AM:

Arbuscular mycorrhizae

AMF:

AM fungi

FWC:

Field water capacity

GM:

Glomus mosseea

MDA:

Malondialdehyde

PAR:

Photosynthetic active radiation

RT:

Reverse transcription

SSH:

Suppression subtractive hybridization

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Acknowledgments

The project is supported by grants from the National Natural Science Foundation of China (30860224), and partially from the Core Subject-constructing Program of Third Term of 211 Engineering (kst200904), Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and Natural Science Foundation of Guizhou Province (20092076).

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Authors and Affiliations

  1. Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou University, Xiahui Street, Huaxi, Guiyang, 550025, Guizhou, People’s Republic of China

    Guang Qiao & Xiaopeng Wen

  2. School of Forestry Science, Guizhou University, Guiyang, 550025, Guizhou, People’s Republic of China

    Lifei Yu

  3. School of Life Science, Guizhou University, Guiyang, 550025, Guizhou, People’s Republic of China

    Xiangbiao Ji

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  1. Guang Qiao
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  2. Xiaopeng Wen
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  3. Lifei Yu
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  4. Xiangbiao Ji
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Correspondence to Xiaopeng Wen.

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Communicated by T. Moriguchi.

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Qiao, G., Wen, X., Yu, L. et al. Identification of differentially expressed genes preferably related to drought response in pigeon pea (Cajanus cajan) inoculated by arbuscular mycorrhizae fungi (AMF). Acta Physiol Plant 34, 1711–1721 (2012). https://doi.org/10.1007/s11738-012-0966-2

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