유전자총 실험조건 최적화를 통한 형질전환 백합 식물체 생산
Optimization of a protocol for the production of transgenic lily plants via particle bombardment
1
조회
0
다운로드
5
인용
0
참고문헌
키워드
초록
Transgenic lily plants have been obtained after particle bombardment, using PDS-1000/He system and scale explants of lilies, followed by PPT (D-L-phosphinothricin) selection. In this study, scales of the lily plants cv. ‘red flame’ were bombarded with a plasmid containing the bar gene as a selectable marker, and the AtSIZ gene as a gene of interest, showing salt tolerance and drought tolerance respectively, and both being driven by the CaMV 35S promoter. For optimization of a protocol, factors which optimized and showed a high transformation efficiency under following conditions, were considered: a bombardment pressure of 1100 psi, a target distance of 6 cm and 1.0 μm of gold particle, and 24-h pre-culture and post-culture on MS medium containing 0.2 M sorbitol and 0.2 M mannitol as osmoticum agents. After bombardment, all the bombarded scales of lily were transferred to MS medium without selective agents, for a week. Subsequently, these bombarded scales were transferred to a selection MS medium containing 10 mg/l PPT, and incubated for a month for further selection, after which they were cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. After transferring into hormone-free MS medium, the PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets. PCR analysis revealed that the surviving putatively transformed plantlets indicated the presence of both the bar gene and the AtSIZ gene. In conclusion, when 100 scales of lily cv. Red flame are bombarded, this study produced approximately 17-18 transgenic plantlets with an optimized bombardment protocol. The protocol described here can contribute to the breeding program of lilies.
Abstract
Transgenic lily plants have been obtained after particle bombardment, using PDS-1000/He system and scale explants of lilies, followed by PPT (D-L-phosphinothricin) selection. In this study, scales of the lily plants cv. ‘red flame’ were bombarded with a plasmid containing the bar gene as a selectable marker, and the AtSIZ gene as a gene of interest, showing salt tolerance and drought tolerance respectively, and both being driven by the CaMV 35S promoter. For optimization of a protocol, factors which optimized and showed a high transformation efficiency under following conditions, were considered: a bombardment pressure of 1100 psi, a target distance of 6 cm and 1.0 μm of gold particle, and 24-h pre-culture and post-culture on MS medium containing 0.2 M sorbitol and 0.2 M mannitol as osmoticum agents. After bombardment, all the bombarded scales of lily were transferred to MS medium without selective agents, for a week. Subsequently, these bombarded scales were transferred to a selection MS medium containing 10 mg/l PPT, and incubated for a month for further selection, after which they were cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. After transferring into hormone-free MS medium, the PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets. PCR analysis revealed that the surviving putatively transformed plantlets indicated the presence of both the bar gene and the AtSIZ gene. In conclusion, when 100 scales of lily cv. Red flame are bombarded, this study produced approximately 17-18 transgenic plantlets with an optimized bombardment protocol. The protocol described here can contribute to the breeding program of lilies.
참고문헌 (33)
- Azadi P, "Macro elements in inoculation and co-cultivation medium strongly affect the efficiency of Agrobacterium-mediated transformation in Lilium", Plant Cell Tiss Organ Cult, 101, pp.201~209 (2010)
- Bakhshaie M, "Biotechnological advance in Lilium", Plant Cell Rep, 35, pp.1799~1826 (2016)
- Deng XY, "Transgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system", Cell Res, 11, pp.156~160 (2001)
- Gao G, "Comparative analysis of transgenic tall fescue (Festuca arundinacea Schreb.) plants obtained by Agrobacterium-mediated transformation and particle bombardment", Plant Cell Rep, 27, pp.1601~1609 (2008)
- Hiei Y, "Efficient transformation of rice(Oryza sativa L. )mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA", Plant J, 6, pp.271~282 (1994)
- Hiei Y, "Transformation of rice mediated by Agrobacterium tumefaciens", Plant Mol Biol, 35, pp.205~218 (1997)
- Hoshi Y, "Production of transgenic lily plants by Agrobacteriummediated transformation", Plant Cell Rep, 22, pp.359~364 (2004)
- Hoshino Y, "Plant regeneration from suspension cells induced from hypocotyls derived from interspecific cross Alstroemeria pelegrina A. magenta and transformation with Agrobacterium tumefaciens", Plant Cell Tiss Organ Cult, 94, pp.45~54 (2008)
- Irifune K, "Production of herbicide resistant transgenic lily plants by particle bombardment", J Jpn Soc Hort Sci, 72, pp.511~516 (2003)
- Kamo K, "Biolistic-mediated transformation of Lilium longiflorum cv. Nellie white", Hortscience, 43(6), pp.1864~1869 (2008)
- Kim JB, "Efficient production of transgenic Alstroemeria plants by using Agrobacterium tumefaciens", Ann Appl Biol, 151, pp.401~412 (2007)
- Langeveld SA, "Transformation of lily by Agrobacterium", Euphytica, 85, pp.97~100 (1995)
- Li SH, "Osmotic sucrose enhancement of single-cell embryogenesis and transformation efficiency in Oncidium", Plant Cell Tiss Organ Cult, 81, pp.183~192 (2005)
- Lin HS, "Genetic transformation of Alstroemeria using particle bombardment", Mol Breed, 6, pp.369~377 (2000)
- Liu J, "Regeneration and production of transgenic Lilium longiflorum via Agrobacterium tumefaciens", Vitro Cell Dev Biol-Plant, 47, pp.348~356 (2011)
- Mercuri A, "Agrobacterium –mediated transformation with rol genes of Lilium longiflorum Thunb", Acta Hort, 612, pp.129~136 (2003)
- Ministry of Food, Agriculture, Forestry and Fisheries, "Statistics of floriculture cultivation in 2015", ~ (2015)
- Morrish F, "Transgenic plants: fundamentals and applications", pp.131~171 (1993), Marcel Dekker
- Murashige T, "A revised medium for rapid growth and bioassays with tobacco tissue culture", Physiol Planta, 15, pp.473~497 (1962)
- 남상욱, "Particle Bombardment에 의한 전처리 된 참나리(Lilium lancifolium Thunb.) 캘러스의 Agrobacterium tumefaciens을 통한 형질전환", Journal of Plant Biotechnology, 31(1), pp.13~17 (2004), 한국식물생명공학회
- Ogaki M, "Importance of co-cultivation medium pH for successful Agrobacterium-mediated transformation of Lilium", Plant Cell Rep, 27, pp.699~705 (2008)
- Park NB, "Proceedings of symposium on the commercialization of lily production and breeding. Hort. Res. Institute RDA", 68~97 (1997)
- Popowich EA, "Agrobacterium-mediated transformation of Hyacinthus orientalis with thaumatin II gene to control fungal disease", Plant Cell Tiss Organ Cult, 90, pp.237~244 (2007)
- 노희선, "유전자총을 이용한 형질전환 심비디움 식물체 생산체계 최적화", Journal of Plant Biotechnology, 38(4), pp.293~300 (2011), 한국식물생명공학회
- 노희선, "심비디움과 팔레놉시스에서 아그로박테리움과 유전자총을 이용한 형질전환효율 비교", 화훼연구, 21(4), pp.199~205 (2013), 한국화훼학회[DOI]
- 노희선, "유전자총을 이용한 팔레놉시스 형질전환 효율향상에 삼투압 조절제 및 발사횟수차이가 미치는 영향", Journal of Plant Biotechnology, 41(4), pp.216~222 (2014), 한국식물생명공학회
- Sanford JC, "Optimizing the biolistic process for different biological applications", Methods Enzymol, 217, pp.483~510 (1993)
- Vain P, "Osmoticum treatment enhances particle bombardment-mediated transient and stable transformation of maize", Plant Cell Rep, 12, pp.84~88 (1993)
- Wang Y, "Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars", Plant Cell Tiss Organ Cult, 111, pp.113~122 (2012)
- Watad AA, "Microprojectile bombardmentmediated transformation of Lilium longiflorum", Plant Cell Rep, 17, pp.262~267 (1998)
- Wilmink A, "Expression of the GUS-gene in the monocot tulip after introduction by particle bombardment and Agrobacterium", Plant Cell Rep, 11, pp.76~80 (1992)
- Woodcock HBD, "Lilies of the world" (1950), Scribners
- Yang J, "Genetic transformation of Cymbidium orchid by particle bombardment", Plant Cell Rep, 18, pp.978~984 (1999)