Interaction of Subculture Cycle, Hormone Ratio, and Carbon Source Regulates Embryonic Differentiation of Somatic Cells in Pinus koraiensis

Abstract

During somatic embryogenesis of Pinus koraiensis, the ability of the embryogenic callus to produce embryos gradually decreases with long-term proliferative culture, which seriously affects large-scale application of this technology. In this study, embryogenic calli of Korean pine 1–100 cell line were used as materials. It was found that in the 7-day subculture cycle of embryogenic calli the somatic embryogenic potential of Korean pine remained for the longer time. In comparison with a subculture cycle of 14 days, indoleacetic acid (IAA), soluble sugar, and starch contents in embryogenic callus were higher with a subculture cycle of 7 days, while hydrogen peroxide (H2O2) content showed the opposite trend. Further, low levels of endogenous abscisic acid (ABA) and high levels of IAA could inhibit somatic embryogenesis of P. koraiensis calli. Among different carbon sources, maltose produced more storage materials and higher superoxide dismutase (SOD) and catalase (CAT) activities than sucrose, which was conducive to cell differentiation and somatic embryogenesis. The results revealed the physiological characteristics of the loss of embryogenic capacity of Korean pine embryogenic callus and provided a basis for the long-term maintenance of its somatic embryogenic capacity.