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Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus

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Abstract

The 5S and 18S rDNA sequences of Catharanthus roseus cv ‘Nirmal’ (wild type) and its leafless inflorescence (lli), evergreen dwarf (egd) and irregular leaf lamina (ill) single mutants and lli egd, lli ill and egd ill double mutants were characterized. The lli, egd and ill mutants of Mendelian inheritance bore the names after their most conspicuous morphological feature(s). They had been chemically induced and isolated for their salt tolerance. The double mutants were isolated as morphological segregants from crosses between single mutants. The morphological features of the two parents accompanied salt tolerance in the double mutants. All the six mutants were hypomethylated at repeat sequences, upregulated and downregulated for many genes and carried pleiotropic alterations for several traits. Here the 5S and 18S rDNAs of C. roseus were found to be relatively low in cytosine content. Cytosines were preponderantly in CG context (53%) and almost all of them were methylated (97%). The cytosines in CHH and CHG (where H = A, T or C) contexts were largely demethylated (92%) in mutants. The demethylation was attributable to reduced expression of RDR2 and DRM2 led RNA dependant DNA methylation and CMT3 led maintenance methylation pathways. Mutants had gained some cytosines by substitution of C at T sites. These perhaps arose on account of errors in DNA replication, mediated by widespread cytosine demethylation at CHG and CHH sites. It was concluded that the regulation of cytosine methylation mechanisms was disturbed in the mutants. ILL, EGD and LLI genes were identified as the positive regulators of other genes mediating the RdDM and CMT3 pathways, for establishment and maintenance of cytosine methylation in C. roseus.

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Acknowledgements

Indian National Science Academy and Council of Scientific and Industrial Research are thanked for grant of scientistship schemes to SK, Department of Biotechnology is thanked for research grant for work on periwinkle and for postgraduate fellowship to RK, Director of NIPGR is thanked for providing research facilities and SKA Institution for Research, Education and Development is thanked for grant of a postgraduate fellowship to VS.

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  1. Genetical Genomics Laboratory, National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi, 110 067, India

    RENU KUMARI, GITANJALI YADAV, VISHAKHA SHARMA & SUSHIL KUMAR

  2. SKA Institution for Research, Education and Development (SKAIRED), 4/11 Sarv Priya Vihar, New Delhi, 110 016, India

    VISHAKHA SHARMA & SUSHIL KUMAR

  3. Banasthali University, P.O. Banasthali Vidhyapeeth, Banasthali, 304 022, India

    RENU KUMARI & VINAY SHARMA

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  1. RENU KUMARI
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[Kumari R., Yadav G., Sharma V., Sharma V. and Kumar S. 2013 Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus. J. Genet. 92, xx–xx]

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KUMARI, R., YADAV, G., SHARMA, V. et al. Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus . J Genet 92, 499–511 (2013). https://doi.org/10.1007/s12041-013-0300-9

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