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Green tissue-specific co-expression of chitinase and oxalate oxidase 4 genes in rice for enhanced resistance against sheath blight

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Abstract

Main conclusion

Green tissue-specific simultaneous overexpression of two defense-related genes ( OsCHI11 & OsOXO4 ) in rice leads to significant resistance against sheath blight pathogen ( R. solani ) without distressing any agronomically important traits.

Overexpressing two defense-related genes (OsOXO4 and OsCHI11) cloned from rice is effective at enhancing resistance against sheath blight caused by Rhizoctonia solani. These genes were expressed under the control of two different green tissue-specific promoters, viz. maize phosphoenolpyruvate carboxylase gene promoter, PEPC, and rice cis-acting 544-bp DNA element, immediately upstream of the D54O translational start site, P D54O–544 . Putative T0 transgenic rice plants were screened by PCR and integration of genes was confirmed by Southern hybridization of progeny (T1) rice plants. Successful expression of OsOXO4 and OsCHI11 in all tested plants was confirmed. Expression of PR genes increased significantly following pathogen infection in overexpressing transgenic plants. Following infection, transgenic plants exhibited elevated hydrogen peroxide levels, significant changes in activity of ROS scavenging enzymes and reduced membrane damage when compared to their wild-type counterpart. In a Rhizoctonia solani toxin assay, a detached leaf inoculation test and an in vivo plant bioassay, transgenic plants showed a significant reduction in disease symptoms in comparison to non-transgenic control plants. This is the first report of overexpression of two different PR genes driven by two green tissue-specific promoters providing enhanced sheath blight resistance in transgenic rice.

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Abbreviations

OA:

Oxalic acid

OXO:

Oxalate oxidase

ROS:

Reactive oxygen species

WT:

Wild type

PDI:

Percentage disease index

DPI:

Days post-infection

PDA:

Potato dextrose agar

APX:

Ascorbate peroxidase

POD:

Peroxidase

SOD:

Superoxide dismutase

CAT:

Catalase

MDA:

Malondialdehyde

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Acknowledgments

This work was funded by the Department of Biotechnology (DBT), Government of India. Kutubuddin Ali Molla is thankful to Council of Scientific and Industrial Research (CSIR) for fellowship. Financial assistance from Indian Council of Agricultural Research (ICAR), Government of India is also acknowledged. Thanks to Dr G.S. Laha for providing R. solani isolates.

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Authors and Affiliations

  1. Laboratory of Translational Research on Transgenic Crops, Department of Botany, University of Calcutta, 35 Ballygunge Circular Road, Kolkata, 700019, West Bengal, India

    Subhasis Karmakar, Kutubuddin Ali Molla, Palas K. Chanda, Sailendra Nath Sarkar, Swapan K. Datta & Karabi Datta

  2. Crop Improvement Division, Central Rice Research Institute, Cuttack, 753006, Odisha, India

    Kutubuddin Ali Molla

  3. Center for Diabetes Research, The Methodist Hospital Research Institute, 6670 Bertner, Houston, TX, 77030, USA

    Palas K. Chanda

  4. Visva Bharati University, Santiniketan, India

    Swapan K. Datta

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  1. Subhasis Karmakar
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  2. Kutubuddin Ali Molla
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Correspondence to Karabi Datta.

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Karmakar, S., Molla, K.A., Chanda, P.K. et al. Green tissue-specific co-expression of chitinase and oxalate oxidase 4 genes in rice for enhanced resistance against sheath blight . Planta 243, 115–130 (2016). https://doi.org/10.1007/s00425-015-2398-x

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