A hypocotyl-derived somatic embryogenic system in Brassica juncea Czern & Coss and its manipulation for enhanced storage lipid accumulation

Abstract

A simple and reproducible protocol for induction, growth and development of somatic embryos from hypocotyl explants of Indian mustard (Brassica juncea L. Czern & Coss) var. RLM 198 is reported. The HDSE (Hypocotyl-derived somatic embryos) were fleshy globular to torpedo structures that were maintained by regular subculturing every three weeks. These embryos developed non-synchronously into the heart shaped-stage while some were matured into a green cotyledon-stage bearing embryos in the same medium. The HDSE accumulated as much as 50.2% lipid content on a dry weight basis at 14 DAC (days after culture) using a culture medium supplemented with 10% PEG (Polyethylene glycol 6000) in comparison to less than 15% lipid content in 2% sucrose (control) or 20 μM ABA (abscisic acid). An increase in total soluble sugar content was observed with 2.5% PEG and increasing PEG concentration caused a decrease in their contents in HDSE. The activities of invertase, acetyl CoA carboxylase and 1-14C-acetate incorporation into lipids in HDSE were enhanced significantly in the culture medium containing 10% PEG. The content of triacylglycerols in HDSE was maximum with 10% PEG supplemented culture medium. The wax content in HDSE increased progressively with an increase of PEG concentration in the culture medium. The ABA and PEG supplementation increased the content of membrane lipids when the data was expressed on a 100 g dry weight basis. The proportion of palmitate and erucate decreased and that of oleate, linoleate and linolenate increased at 14 DAC in HDSE in 10% PEG supplemented culture medium. Thus, the manipulation of culture conditions significantly altered total lipid content, membrane lipid composition and the quality of storage lipids.