Browsing by Author "Taneja, Jyoti"

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    Cloning and characterization of two new Class III peroxidase genes from Catharanthus roseus
    (Elsevier, 2011) Kumar, Santosh; Jaggi, Monika; Taneja, Jyoti; Sinha, Alok Krishna
    We report here the cloning and characterization of two new class III peroxidase genes, CrPrx3 and CrPrx4 from Catharanthus roseus. The full length cDNA of CrPrx3 is 1233 bp long encoding 330 amino acid residues. The CrPrx4 cDNA contains an ORF of 1055 bp, and encodes for 318 amino acids. Hypothetical 3-D structure model of CrPrx3 and CrPrx4 detected the presence of two Ca(2+) ion binding sites at proximal and distal end, and a heme group coordinate at central position. CrPrx3 and CrPrx4 are present in single copy number in C. roseus. Confocal scanning microscopy and phylogenetic analysis showed that CrPrx3 and CrPrx4 are apoplastic in nature. Phylogenetic analysis revealed that both genes have diverse expression patterns in a variety of plant tissues. Quantitative real-time PCR confirmed their maximum expression in stem tissues followed by flower tissue. The expression levels of CrPrx3 and CrPrx4 were down-regulated under salt and dehydration stress, respectively.
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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
    (Springer, 2010) Taneja, Jyoti; Jaggi, Monika; Wankhede, Dhammaprakash Pandhari; Sinha, Alok Krishna
    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 T(R)-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.