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Functional screening of cDNA library from a salt tolerant rice genotype Pokkali identifies mannose-1-phosphate guanyl transferase gene (OsMPG1) as a key member of salinity stress response

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Abstract

Salinity, one of the most deleterious stresses, affects growth and overall yield of crop plants. To identify new “candidate genes” having potential role in salinity tolerance, we have carried out ‘functional screening’ of a cDNA library (made from a salt tolerant rice—Pokkali). Based on this screening, we identified a cDNA clone that was allowing yeast cells to grow in the presence of 1.2 M NaCl. Sequencing and BLAST search identified it as mannose-1-phosphate guanyl transferase (OsMPG1) gene from rice. Analysis of rice genome sequence database indicated the presence of 3 additional genes for MPG. Out of four, three MPG genes viz. OsMPG1, 3 and 4 were able to functionally complement yeast MPG mutant -YDL055C. We have carried out detailed transcript profiling of all members of MPG family by qRT-PCR using two contrasting rice genotypes (IR64 and Pokkali) under different abiotic stresses (salinity, drought, oxidative stress, heat stress, cold or UV light). These MPG genes showed differential expression under various abiotic stresses with two genes (OsMPG1 and 3) showing high induction in response to multiple stresses. Analysis of rice microarray data indicated higher expression levels for OsMPG1 in specific tissues such as roots, leaves, shoot apical meristem and different stages of panicle and seed development, thereby indicating its developmental regulation. Functional validation of OsMPG1 carried out by overexpression in the transgenic tobacco revealed its involvement in enhancing salinity stress tolerance.

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Acknowledgments

Authors are thankful for the funds received from International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi and Department of Biotechnology (DBT) and Department of Science & Technology (DST), Government of India. AM thanks ICGEB for providing post-doctoral fellowship.

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Authors and Affiliations

  1. Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Ritesh Kumar, Ananda Mustafiz, Khirod Kumar Sahoo, Vishal Sharma, Subhasis Samanta, Sudhir Kumar Sopory & Sneh Lata Singla-Pareek

  2. Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Ashwani Pareek

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  1. Ritesh Kumar
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  2. Ananda Mustafiz
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  3. Khirod Kumar Sahoo
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  4. Vishal Sharma
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  5. Subhasis Samanta
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  6. Sudhir Kumar Sopory
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  7. Ashwani Pareek
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Correspondence to Sneh Lata Singla-Pareek.

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Ritesh Kumar and Ananda Mustafiz contributed equally to this paper.

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11103_2012_9928_MOESM2_ESM.ppt

Representative picture to show screening of rice cDNA library in yeast system under high salinity. The rectangle marks one of the yeast clones showing normal robust growth and maintaining its rounded morphology on the medium containing 900 mM NaCl, while all other yeast clones showed scattered and irregular growth on this media. (PPT 222 kb)

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Kumar, R., Mustafiz, A., Sahoo, K.K. et al. Functional screening of cDNA library from a salt tolerant rice genotype Pokkali identifies mannose-1-phosphate guanyl transferase gene (OsMPG1) as a key member of salinity stress response. Plant Mol Biol 79, 555–568 (2012). https://doi.org/10.1007/s11103-012-9928-8

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