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Lipids in hairy roots and non-Agrobacterium induced roots of Crambe abyssinica

Głąb, Bartosz, Furmanek, Tomasz, Miklaszewska, Magdalena, Banaś, Antoni, Królicka, Aleksandra
Acta physiologiae plantarum 2013 v.35 no.7 pp. 2137-2145
Crambe abyssinica, Rhizobium rhizogenes, clones, free fatty acids, genes, leaves, linoleic acid, lipid content, palmitic acid, polymerase chain reaction, roots, sterol esters, transfer DNA, triacylglycerols
Hairy root cultures of Crambe abyssinica were obtained through infection of leaves with two wild-type agropine strains of Agrobacterium rhizogenes. The efficiency of transformation was about 16 %. The presence of T-DNA from A. rhizogenes in the hairy roots genome was confirmed by PCR using specific primers for rolB and rolC genes. Selected clones of hairy roots and non-Agrobacterium induced roots from sterile cultures were used for analyses of acyl-lipids. The total amount of acyl-lipids per mg of dry weight was similar in both the non-Agrobacterium induced roots and the hairy roots in good physiological condition, and ranged from 38 to 53 nmol. However, in the clones which showed symptoms of ageing, the lipid content was severely reduced. Also the lipid composition of hairy roots appears to be similar to the composition of non-transformed roots. Polar lipids were the dominant class of lipids in both types of roots (about 75 %). Furthermore, we found diacylglycerols, free fatty acids (FFA), triacylglycerols, sterol esters, and an unidentified lipid class. The dominant fatty acids in the lipids of both types of roots were α-linolenic acid, palmitic acid, and linoleic acid (over 12 % of total FA). Among the lipids of both hairy roots and non-Agrobacterium induced roots of C. abyssinica, an unidentified FA was found (over 16 % of total FAs). The present study is the first example of establishment of hairy roots cultures of C. abyssinica. It also includes the first analysis of the lipids in hairy roots and non-Agrobacterium induced roots of this species.