Efficient Technique for Long-Term in Vitro Storage of Transgenic Aspen Genotypes - American Journal of Plant Sciences

AJPS > Vol.9 No.13, December 2018

Efficient Technique for Long-Term in Vitro Storage of Transgenic Aspen Genotypes ()

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Download as PDF (Size: 591KB) PP. 2593-2600

DOI: 10.4236/ajps.2018.913188 654 Downloads 2,077 Views Citations

Author(s)

Elena O. Vidyagina^*, Konstantin A. Shestibratov

Affiliation(s)

Branch of the Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Pushchino, Russia.

ABSTRACT

In vitro culture of isolated cells from tissues and organs is sometimes used to preserve and reproduce unique genotypes of woody plants. The technique, however, requires regular subculturing which raises storage costs and creates risks for contamination and accumulation of somaclonal variations. We examined the effects of sugar composition of culture medium, the length of photoperiod, light intensity, and ambient temperature on the survival of plant material in vitro. The study was performed on 49 genotypes of Populus tremula (46 transgenic genotypes carrying GFP-, Xeg- and Gus-genes, and 3 control (wild-type) genotypes). It was shown that effective storage of plants was achieved through optimization of the combined effects of all storage parameters under study. Based on the experimental data, we developed a protocol for long-term in vitro storage of desirable genotypes without subculture and with a survival rate of up to 98%. The best results were obtained when the plant material was pre-cultured on a WPM medium containing 15 g/L sucrose, 7.5 g/L sorbitol and 7.5 g/L mannitol, and then stored at +4°C under a 24-hour light day cycle with only 8 hours of light per day and maximum light intensity of 2000 lux. Post-storage recovery was done by culturing on a medium containing 1 mg/L gibberellic acid. The developed method can be used for effective in vitro storage of the studied genotypes for up to 24 months without subculture.

KEYWORDS

Populus tremula, Long-Term Storage, In Vitro Culture, Osmolytics, Transgenic Genotypes, Microshoots

Share and Cite:

Vidyagina, E. and Shestibratov, K. (2018) Efficient Technique for Long-Term in Vitro Storage of Transgenic Aspen Genotypes. American Journal of Plant Sciences, 9, 2593-2600. doi: 10.4236/ajps.2018.913188.

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