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Effect of loss of T-DNA genes on MIA biosynthetic pathway gene regulation and alkaloid accumulation in Catharanthus roseus hairy roots

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Abstract

Hairy roots are generated by integration of T-DNA in host plant genome from root inducing (Ri) plasmid of Agrobacterium rhizogenes and have been utilized for production of secondary metabolites in different plant systems. In Catharanthus roseus, hairy roots are known to show different morphologies, growth patterns, and alkaloid contents. It is also known that during transformation, there is a differential loss of a few T-DNA genes. To decipher the effect of loss of T-DNA genes on the various aspects of hairy roots, ten hairy root clones were analyzed for the presence or absence of T-DNA genes and its implications. It was found that the loss of a few ORFs drastically affects the growth and morphological patterns of hairy roots. The absence of TR-DNA from hairy roots revealed increased transcript accumulation and higher alkaloid concentrations, whereas callusing among hairy root lines led to decreased transcript and alkaloid accumulation. Significantly higher expression of MIA biosynthetic pathway genes and low abundance of regulator transcripts in hairy root clones in comparison with non-transformed control roots were also observed. This study indicates that it is not only the integration of T-DNA at certain region of host plant genome but also the presence or absence of important ORFs that affects the expression patterns of MIA biosynthetic pathway genes, regulators, and accumulation of specific alkaloids.

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Abbreviations

MIAs:

Monoterpenoid indole alkaloids

Ri:

Root inducing plasmid

rol:

Root locus

T-DNA:

Transferred DNA

TL-DNA:

Left transferred DNA

TR-DNA:

Right transferred DNA

dw:

Dry weight

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Acknowledgments

JB and MJ thank Council of Scientific and Industrial Research (CSIR), India while DPW thanks University Grant Commission, India for the award of senior research fellowships. This work is supported by financial assistance from the core grant of National Institute of Plant Genome Research, New Delhi, India.

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  1. National Institute of Plant Genome Research, P. O. Box 10531, Aruna Asaf Ali Marg, New Delhi, 110 067, India

    Jyoti Taneja, Monika Jaggi, Dhammaprakash Pandhari Wankhede & Alok Krishna Sinha

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  1. Jyoti Taneja
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  2. Monika Jaggi
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  3. Dhammaprakash Pandhari Wankhede
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  4. Alok Krishna Sinha
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Correspondence to Alok Krishna Sinha.

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Communicated by P. Lakshmanan.

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Taneja, J., Jaggi, M., Wankhede, D.P. et al. Effect of loss of T-DNA genes on MIA biosynthetic pathway gene regulation and alkaloid accumulation in Catharanthus roseus hairy roots. Plant Cell Rep 29, 1119–1129 (2010). https://doi.org/10.1007/s00299-010-0895-8

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