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The study was conducted to investigate the possibility that epigenetic DNA methylation causes tree phenotypic variation in autotetraploids through evaluating the phenotypic variation and DNA methylation in autotetraploids occurred spontaneously from diploid trifoliate orange. Chromosome analysis confirmed that fourteen trifoliate orange trees of selected by flow cytometry were tetraploids (2n = 4X = 36) without any aneuploids. Chromomycin A3 staining determined that these trees were all autotetraploid with doubled chromosome set. Tree phenotypes, such as tree height and width, branching number, length, and angle, internode length, and leaf characteristics, varied in the autotetraploids. Chlorophyll indices were diverse in the autotetraploids, but photosynthetic rates were not significantly different. In addition, a wide range of variation was observed in stomatal density and guard cell length. Analysis of global cytosine DNA methylation showed that there was a variation of the methylation level in autotetraploids. More than half of 14 autotetraploids had at least 2 times higher methylation level than diploid trifoliate orange. The results indicate that tree phenotypic variation in autotetraploids might be related to global DNA methylation for reducing gene redundancy. Additional key words: chromosome doubling, CMA karyotyping, DNA methylation, epigenetics *Corresponding author: kwansong@jejunu.ac.kr ※ Received 29 August 2013; Revised 6 December 2013; Accepted 5 February 2014. This work was carried out with the support of “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ00961202)” Rural Development Administration, Republic of Korea. C 2014 Korean Society for Horticultural Science           (FAO, 2012; MFAFF, 2011).       ,   ,  ,               .  , ,         .                  .         .                  (Recupero et al., 2005).     , , , Jian Liang Lu, , ,  367              (Zhu et al., 2009).            (Ferrante et al., 2009; Recupero et al., 2005).     (2n = 18) .         .        (Dutt et al., 2010; Wakana et al., 2005),      (Grosser et al., 1994, 2000; Medina-Urrutia et al., 2004)      (Ferrante et al., 2009; Kaneyoshi et al., 2008; Oiyama and Kobayashi, 1991; Song et al., 2011; Usman et al., 2006)   .                  (Cameron and Frost, 1968).     2-5%   .        ,       .   ,   ,      .         (Cameron and Frost, 1968).           (Romero-Aranda et al., 1997; Wakana et al., 2005),             .             ,         (Gunasekare and Ranatunga, 2003; Osborn et al., 2003),   (Ranatunga and Gunasekare, 2003; Ye et al., 2010).                      ,  ,          (Osborn et al., 2003; Sémon and Wolfe, 2007).           ,  ,      .                  (Osborn et al., 2003; Sémon and Wolfe, 2007).                                       .

Chromosome Redundancy and Tree Phenotype Variation in Autotetraploid Trifoliate Orange