Crambe aspera plants in vitro propagation and its effect on fatty acids and phenolic compounds content and genome stability

Abstract

In vitro culture can be used for endangered species preservation but its effect on biochemical properties and genetic stability of plants requires further research. Aim. The study on fatty acids content, phenolic compounds and phylogenetic relationships of Crambe aspera plants upon aseptic cultivation in vitro. Methods. Morphogenic potential of Crambe aspera leaf, root and petiole explants was studied in the Murashige and Skoog (MS) medium with different concentrations of growth factors. Fatty acids (FA) content was determined by gas chromatography-mass spectrophotometry of FA ethers, phenolic compounds were determined by spectrophotometry. Polymerase chain reactions were used to study phylogenetic relationships. Results. The efficient protocols of seed surface sterilization as well as methods of fast microclonal multiplication and obtention of C. aspera callus tissue for were developed. Conclusions. Leaves of both in vitro and in vivo cultured plants had a high content of α-linolenic acid whereas erucic acid was absent. At the same time, the difference in biochemical composition between the plants grown in aseptic and non-aseptic conditions was shown. In vivo populations of C. aspera showed a high level of polymorphism but its genome did not undergo changes after the in vitro establishment.

Biotechnological approach for improvement of Crambe species as valuable oilseed plants for industrial purposes

“…This research was performed to estimate the possible use of root, leaf and petiole explants for in vitro propagation of Crambe plants that are endangered in Ukraine, as well as in other counties, and to obtain the basis for Crambe gene pool improvement. 64–68,78…”

“…The highest propagation rates for C. koktebelica petiole explants (60%) were noted on the medium with 2.5–5 mg L −1 of BA and 0.1 mg L −1 of NAA (Table 1). 64–68,78…”

“…C. maritima, C. grandiora, C.cordifolia, C. mitridatis). [64][65][66][67][68] It is necessary to mention that, although pericarp removal can be easily performed at an industrial scale, it could increase the risk of seed damage and spreading of fungal infection, so it should be performed when the risk is justied.…”

“…Seed surface sterilization is a way to obtain aseptic plant culture in the least damaging way for the plant material. The highest number of aseptic plantlets was obtained from the seeds surface sterilization with diocidum (the exposure time 2-3 minutes) [64][65][66][67][68] or commercial bleach solution (exposure time 10-20 minutes). [69][70][71] Also, 3% hydrogen peroxide for 10 minutes was successfully used for that purpose.…”

“…71 Aer the seed surface sterilization seeds were transferred to culture medium of choice (mainly MS or half strength MS) in culture chamber at 22-25 C with 16-hour photoperiod. [64][65][66][67][68][69][70][71] Time of Crambe aseptic seed germination aer the seed surface sterilization procedure varied and lasted from 3 days to a month depending on the species, seed storage time and conditions, and on the seed coat removal. [64][65][66][67][68][69][70][71][72][73] For aseptic culture establishment explants from aseptic seedlings were taken and transferred to a medium for further cultivation and micropropagation.…”