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Optimized protoplast isolation and establishment of transient gene expression system for the Antarctic flowering plant Colobanthus quitensis (Kunth) Bartl.

Cited 2 time in wos
Cited 3 time in scopus
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Title
Optimized protoplast isolation and establishment of transient gene expression system for the Antarctic flowering plant Colobanthus quitensis (Kunth) Bartl.
Other Titles
남극현화식물인 남극개미자리의 유전자발현시스템을 위한 원형질분리법
Authors
Cha, Ok-Kyoung
Lee, Jong Eun
Lee, Hyoungseok
Lee, Horim
Subject
Biotechnology & Applied Microbiology; Plant Sciences
Keywords
Cell-based assay; Low temperature; Molecular adaptation; PEG-CaCl2-mediated transfection; Subcellular localization
Issue Date
2019-09
Citation
Cha, Ok-Kyoung, et al. 2019. "Optimized protoplast isolation and establishment of transient gene expression system for the Antarctic flowering plant Colobanthus quitensis (Kunth) Bartl.". PLANT CELL TISSUE AND ORGAN CULTURE, 138(3): 603-607.
Abstract
Colobanthus quitensis is one of two terrestrial plants that grow in the maritime Antarctic. Despite its important ecological niche in extreme environments, the molecular mechanisms of its adaptation and tolerance have not been elucidated due to difficulties with genetic or transgenic approaches. For this reason, in many other plant species mesophyll protoplasts as a versatile cell-based system have been developed and used to analyze the biological functions of genes of interest. Here we report an optimized method of protoplast isolation from C. quitensis leaves. The main parameters evaluated to reach the highest protoplast yield were the use of a cell wall-degrading enzyme, an osmotic stabilizer, and different pH conditions. Moreover, transient expression and subcellular localization of proteins were validated by an immunoblot assay and a confocal microscopy, respectively, using C. quitensis protoplasts. Therefore, these results suggest that protoplasts can provide a useful cell-based system to facilitate the molecular, biochemical, and cellular characterizations of C. quitensis genes.
URI
https://repository.kopri.re.kr/handle/201206/10933
DOI
http://dx.doi.org/10.1007/s11240-019-01651-1
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