Abstract
In present study, methylation-sensitive AFLP (MSAP) markers were employed to assess DNA methylation, degree of alterations in DNA methylation and methylation polymorphism in plant tissues growing in vivo and in vitro. The leaf tissues of six plants growing in vivo and in vitro were subjected to MSAP profiling. A total of 717 MSAP markers in Salvadora persica, 801 in Commiphora wightii, 874 in male (M) and 845 in female (F) genotype of Simmondsia chinensis, 719 in Jatropha curcas and 880 in Withania coagulans were obtained with seventeen MSAP primer combinations. Percentage methylation in genome obtained was higher in in vivo-grown tissues of S. persica (39.47 %), S. chinensis—M (61.71 %) and W. coagulans (71.59 %); and in in vitro-grown tissues of C. wightii (65.17 %), S. chinensis—F (60.83 %) and J. curcas (68.29 %). The percentage polymorphism in methylated DNA obtained was 8.71 % in S. persica, 9.81 % in J. curcas, 10.10 % in S. chinensis—F, 10.26 % in W. coagulans, 10.66 % in S. chinensis—M and 13.98 % in C. wightii. The difference in DNA methylation and polymorphism in genomes reflect the plasticity in genomes of the plants growing under two different environments. Different pattern of DNA methylation of the homologous nucleotide sequences and polymorphism in the methylated DNA in tissues under in vitro and in vivo conditions suggest possibility of involvement of these fragments in the dynamic processes regulating plant growth and development under prevailing growth conditions.
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References
Arnholdt-Schmitt B, Herterich S, Neumann K-H (1995) Physiological aspects of genome variability in tissue culture. I. Growth phase-dependent differential DNA methylation of the carrot genome (Daucus carota L.) during primary culture. Theor Appl Genet 91:809–815
Berdasco M, Alcázar R, García-Ortiz MV, Ballestar E et al (2008) Promoter DNA hypermethylation and gene repression in undifferentiated Arabidopsis cells. PLoS One 3(10):e3306
Bottley A, Chapman NH, Koebner RMD (2008) Homoeologous gene silencing in tissue cultured wheat callus. BMC Genet 9:65
Brautigam K, Vining KJ, Lafon-Placette C et al (2013) Epigenetic regulation of adaptive responses of forest tree species to the environment. Ecol Evol 3(2):399–415
Chen L, Chen J (2008) Changes of cytosine methylation induced by wide hybridization and allopolyploidy in Cucumis. Genome 51:789–799
Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15
Fraga MM, Canal MJ, Rodriguez R (2002) Phase-change related epigenetic and physiological changes in Pinus radiata D. Don. Planta 215:672–678
Golyasnaya N, Tsvetkova N (2006) Mismatch repair. Mol Biol 40:183–193
Gonzalez A, Saiz A, Acedo A, Ruiz M, Polanco C (2013) Analysis of genomic DNA methylation patterns in regenerated and control plants of rye (Secale cereale L.). Plant Growth Regul 70(3):227–236
Goodrich J, Tweedie S (2002) Remembrance of things past: chromatin remodeling in plant development. Annu Rev Cell Dev Biol 18:707–746
Habu Y, Kakutani T, Paszkowski J (2001) Epigenetic developmental mechanisms in plants: molecules and targets of plant epigenetic regulation. Curr Opin Genet Dev 11:215–220
Huang H, Han SS, Wang Y, Zhang XZ, Han ZH (2012) Variations in leaf morphology and DNA methylation following in vitro culture of Malus xiaojinensis. Plant Cell Tiss Organ Cult 111(2):153–161
Kaeppler SM, Kaeppler HF, Rhee Y (2000) Epigenetic aspects of somaclonal variation in plants. Plant Mol Biol 43:179–188
Keyte AL, Percifield R, Liu B, Wendel JF (2006) Infraspecific DNA methylation polymorphism in cotton (Gossypium hirsutum L.). J Hered 97(5):444–450
Krizova K, Fojtova M, Depicker A, Kovarik A (2009) Cell culture induced gradual and frequent epigenetic reprogramming of invertedly repeated tobacco transgene epialleles. Plant Physiol 149:1493–1504
Li X, Xu M, Korban SS (2000) DNA methylation profiles differ between field- and in vitro-grown leaves of apple. J Plant Physiol 159(11):1229–1234
Madlung A, Tyagi AP, Watson B et al (2005) Genomic changes in synthetic Arabidopsis polyploids. Plant J 41:221–230
Mastan SG, Rathore MS, Bhatt VD, Yadav P, Chikara J (2012) Assessment of changes in DNA methylation by methylation-sensitive amplification polymorphism in Jatropha curcas L. subjected to salinity stress. Gene 508:125–129
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479
Phillips RL, Kaeppler SM, Olhoft P (1994) Genetic instability of plant tissue cultures: breakdown of normal controls. Proc Natl Acad Sci USA 91:5222–5226
Rapp RA, Wendel JF (2005) Epigenetics and plant evolution. New Phytol 168(1):81–91
Rathore MS, Shekhawat S, Kaur G, Singh RP, Shekhawat NS (2012) Micropropagation of vegetable rennet (Withania coagulans [Stocks] Dunal)—a critically endangered medicinal plant. J Sust Forest 31(8):727–746
Riddle NC, Richards EJ (2002) The control of natural variation in cytosine methylation in Arabidopsis. Genetics 162:355–363
Rival A, Ilbert P, Labeyrie A et al (2013) Variations in genomic DNA methylation during the long-term in vitro proliferation of oil palm embryogenic suspension cultures. Plant Cell Rep 32(3):359–368
Valledor L, Hasbun R, Meijon M et al (2007) Involvement of DNA methylation in tree development and micropropagation. Plant Cell Tissue Organ Cult 91(2):75–86
Vanyushin BF, Ashapkin VV (2011) DNA methylation in higher plants: past, present and future. BBA—Gene Regul Mech 1809(8):360–368
Verhoeven KJF, Jansen JJ, van Dijk PJ, Biere A (2010) Stress-induced DNA methylation changes and their heritability in asexual dandelions. New Phytol 185:1108–1118
Wada Y, Miyamoto K, Kusano T, Sano II (2004) Association between up-regulation of stress-responsive genes and hypomethylation of genomic DNA in tobacco plants. Mol Gen Genomics 271:658–666
Wang Q-M, Wang Y-Z, Sun L-L, Gao F-Z, Sun W, He J, Gao X, Wang L (2012) Direct and indirect organogenesis of Clivia miniata and assessment of DNA methylation changes in various regenerated plantlets. Plant Cell Rep 31:1283–1296
Xu M, Li X, Korban SS (2000) AFLP-based detection of DNA methylation. Plant Mol Biol Rep 18:361–368
Zhang M, Kimatu JN, Xu K, Liu B (2010) DNA cytosine methylation in plant development. Review. J Genet Genomics 37:1–12
Acknowledgments
CSIR-CSMCRI Communication No. 004/2014. Authors are thankful to CSIR (MLP 0014), New Delhi and CSIR-SRF for financial assistance. S. G. Mastan is also thankful to CSIR for SRF; and AcSIR for enrolment in Ph.D.
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Rathore, M.S., Mastan, S.G. & Agarwal, P.K. Evaluation of DNA methylation using methylation-sensitive amplification polymorphism in plant tissues grown in vivo and in vitro. Plant Growth Regul 75, 11–19 (2015). https://doi.org/10.1007/s10725-014-9926-8
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DOI: https://doi.org/10.1007/s10725-014-9926-8