Karim Rezaul, Tan Yew Seong, Singh Pooja, Khalid Norzulaani, Harikrishna Jennifer Ann
1Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
2Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, 50603 Kuala Lumpur, Malaysia.
Physiol Mol Biol Plants. 2018 Sep;24(5):741-751. doi: 10.1007/s12298-018-0566-8. Epub 2018 Jun 28.
The process of somatic embryogenesis and plant regeneration involve changes in gene expression and have been associated with changes in DNA methylation. Here, we report the expression and DNA methylation patterns of - (), (), () and () in meristematic block of newly emerged shoots from rhizome, embryogenic and non-embryogenic calli, prolonged cell suspension culture, ex vitro leaf, and in vitro leaf of regenerated plants of . Among all seven samples, based on qRT-PCR, the highest level of expression of and was in embryogenic callus, while was most highly expressed in meristematic block tissue followed by embryogenic callus. Relatively lower expression was observed in cell suspension culture and watery callus for and and in in vitro leaf for . For gene specific methylation determined by bisulfite sequencing data, embryogenic callus samples had the lowest levels of DNA methylation at CG, CHG and CHH contexts of , and . We observed negative correlation between DNA methylation at the CG and CHG contexts and the expression levels of , , and . Based on our results, we suggest that relatively higher expression and lower level of DNA methylation of , and are associated with somatic embryogenesis and plant regeneration in .
体细胞胚胎发生和植株再生过程涉及基因表达的变化，并且与DNA甲基化的改变有关。在此，我们报告了[植物名称]根茎新抽出芽的分生组织块、胚性愈伤组织和非胚性愈伤组织、长期细胞悬浮培养物、离体叶片以及再生植株的离体叶片中[基因名称1]、[基因名称2]、[基因名称3]和[基因名称4]的表达和DNA甲基化模式。在所有七个样品中，基于qRT-PCR，[基因名称1]和[基因名称2]的最高表达水平出现在胚性愈伤组织中，而[基因名称3]在分生组织块组织中表达最高，其次是胚性愈伤组织。对于[基因名称1]和[基因名称2]，在细胞悬浮培养物和水渍状愈伤组织中观察到相对较低的表达，对于[基因名称3]，在离体叶片中表达相对较低。通过亚硫酸氢盐测序数据确定的基因特异性甲基化方面，胚性愈伤组织样品在[基因名称1]、[基因名称2]和[基因名称3]的CG、CHG和CHH序列背景下具有最低的DNA甲基化水平。我们观察到CG和CHG序列背景下的DNA甲基化与[基因名称1]、[基因名称2]、[基因名称3]和[基因名称4]的表达水平之间呈负相关。基于我们的结果，我们认为[基因名称1]、[基因名称2]和[基因名称3]相对较高的表达和较低水平的DNA甲基化与[植物名称]的体细胞胚胎发生和植株再生有关。
