Comparison of Cryoprotectants and Cryopreservation Protocols for Eleutherococcus senticosus via Somatic Embryogenesis
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
A cryopreservation is an essential tool for preservation of germplasm. In this study, the possibility for cryopreservation of embryogenic cells of Siberian ginseng (Eleutherococcus senticosus) in liquid nitrogen (-196°C) was evaluated. The effects of glycerol and dimethyl sulfoxide (DMSO) at different concentrations (5%, 10% and 20%) as cryoprotectants on regrowth of cryopreserved E. senticosus embryogenic cells were tested. There was significant effect of cryoprotectants on regrowth of embryogenic cells (p=0.0019). The highest and lowest fresh mass gain were achieved when embryogenic cells were frozen with 10% DMSO and 5% glycerol (138.2±5.9 and 61.3±14.6, respectively). The effect of the cryoprotectants on the frequency embryo germination was tested. There was no significant difference between glycerol and DMSO (p=0.846). Three different concentrations of cryoprotectants did not significantly affect the frequency embryo germination (p=0.534). Finally, the genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cells was tested by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. RAPD and ISSR analysises showed that there was no genetic variation among regenerants.
Abstract
A cryopreservation is an essential tool for preservation of germplasm. In this study, the possibility for cryopreservation of embryogenic cells of Siberian ginseng (Eleutherococcus senticosus) in liquid nitrogen (-196°C) was evaluated. The effects of glycerol and dimethyl sulfoxide (DMSO) at different concentrations (5%, 10% and 20%) as cryoprotectants on regrowth of cryopreserved E. senticosus embryogenic cells were tested. There was significant effect of cryoprotectants on regrowth of embryogenic cells (p=0.0019). The highest and lowest fresh mass gain were achieved when embryogenic cells were frozen with 10% DMSO and 5% glycerol (138.2±5.9 and 61.3±14.6, respectively). The effect of the cryoprotectants on the frequency embryo germination was tested. There was no significant difference between glycerol and DMSO (p=0.846). Three different concentrations of cryoprotectants did not significantly affect the frequency embryo germination (p=0.534). Finally, the genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cells was tested by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. RAPD and ISSR analysises showed that there was no genetic variation among regenerants.
참고문헌 (29)
- Ahn CH, "Random amplified polymorphic DNA (RAPD) analysis and RAPD-derived sequence characterized amplified regions (SCAR) marker development to identify Chinese and Korean ginseng", J Med Plants Res, 5, pp.4487~4492 (2011)
- Al-Bahrany AM, "Optimizing in vitro cryopreservation of date palm (Phoenix dactylifera L.)", Biotechnology, 11, pp.59~66 (2012)
- 안찬훈, "In Vitro Propagation of Commonly Used Medicinal Trees in Korea", Journal of forest and environmental science, 35(4), pp.272~280 (2019), 산림과학연구소[DOI]
- Arakawa T, "The basis for toxicity of certain cryoprotectants: a hypothesis", Cryobiology, 27, pp.401~415 (1990)
- Benson EE, "Cryopreservation of phytodiversity : a critical appraisal of theory & practice", Crit Rev Plant Sci, 27, pp.141~219 (2008)
- Bhattacharya S, "Cryopreservation Biotechnology in Biomedical and Biological Sciences", pp.7~20 (2018), IntechOpen
- Chakrabarty D, "Detection of DNA methylation changes during somatic embryogenesis of Siberian ginseng (Eleuterococcus senticosus)", Plant Sci, 165, pp.61~68 (2003)
- CHEONG EUN JU, "Organogenesis from Callus Derived from In Vitro Root Tissues of Wild Prunus yedoensis Matsumura", Journal of forest and environmental science, 35(1), pp.41~46 (2019), 산림과학연구소[DOI]
- Choi YE, "Dormancy induction of somatic embryos of Siberian ginseng by high sucrose concentrations enhances the conservation of hydrated artificial seeds and dehydration resistance", Plant Cell Rep, 20, pp.1112~1116 (2002)
- Choi YE, "High frequency of plant production via somatic embryogenesis from callus or cell suspension cultures in Eleutherococcus senticosus", Annal Bot, 83, pp.309~314 (1999)
- Choi YE, "Production of plantlets of Eleutherococcus sessiliflorus via somatic embryogenesis and successful transfer to soil", Plant Cell Tissue Organ Cult, 69, pp.201~204 (2002)
- Choudhary R, "Genetic stability of mulberry germplasm after cryopreservation by two-step freezing technique", Afr J Biotechnol, 12, pp.5983~5993 (2013)
- Fernandes P, "Cryopreservation of Quercus suber somatic embryos by encapsulation-dehydration and evaluation of genetic stability", Tree Physiol, 28, pp.1841~1850 (2008)
- Saikat Gantait, "Cryopreservation of Forest Tree Seeds: A Mini-Review", Journal of forest and environmental science, 32(3), pp.311~322 (2016), 산림과학연구소
- Gui Y, "Somatic embryogenesis and plant regeneration in Acanthopanax senticosus", Plant Cell Rep, 9, pp.514~516 (1991)
- 한정연, "세포 배양을 통한 가시오갈피 (Eleutherococcus senticosus) 묘목의 대량 생산 시스템 개발", Journal of Plant Biotechnology, 30(2), pp.167~172 (2003), 한국식물생명공학회
- Harding K, "Genetic integrity of cryopreserved plant cells: a review", Cryo Letters, 25, pp.3~22 (2004)
- 정재훈, "가시오갈피 기내 식물체의 생장, 형태형성 및 eleutheroside 함량에 미치는 발광다이오드의 효과", 한국약용작물학회지, 17(1), pp.39~45 (2009), 한국약용작물학회
- Lee WT, "Distribution of Acanthopanax plants in Korea", Korean J Pharmacogn, 10, pp.103~107 (1979)
- Lee YS, "Effects of the water extract from the stem bark of Acanthopanax senticosus on hyperlipidemia in rats", Korean J Pharmacogn, 32, pp.103~107 (2001)
- 이성호, "가시오갈피 현탁배양 체세포배의 저온장기저장 및 식물체 재분화", 한국약용작물학회지, 12(6), pp.494~499 (2004), 한국약용작물학회
- 이성호, "생물반응기에서 배양한 가시오갈피 유식물체의 순화 및 생육특성", 한국약용작물학회지, 13(4), pp.133~137 (2005), 한국약용작물학회
- Mikuła A, "Cryopreservation enhances embryogenic capacity of Gentiana cruciata (L.) suspension culture and maintains (epi)genetic uniformity of regenerants", Plant Cell Rep, 30, pp.565~574 (2011)
- Murashige T, "A revised medium for rapid growth and bio assays with tobacco tissue cultures", Physiol Plant, 15, pp.473~497 (1962)
- Panis B, "The Role of Biotechnology for the Characterization and Conservation of Crop, Forest, Animal and Fishery Genetic Resources in Developing Countries" (2005), FAO
- Richards CM, "Microsatellite primers for Texas wild rice (Zizania texana), and a preliminary test of the impact of cryogenic storage on allele frequency at these loci", Conserv Genet, 5, pp.853~859 (2004)
- Yuwon Seol, "Optimization Conditions for Cryopreservation of Deutzia paniculata Nakai, Endangered Plant", Journal of forest and environmental science, 36(4), pp.274~280 (2020), 산림과학연구소[DOI]
- Sun YL, "Eleutherococcus senticosus as a crude medicine: review of biotechnological effects", J Med Plants Res, 5, pp.6105~6111 (2011)
- Yu CY, "Callus formation and plant regeneration from immature embryos of Eleutherococcus senticosus", Korean J Med Crop Sci, 5, pp.49~55 (1997)