Salt cress 유전자의 형질전환을 통한 내염성 식물체 선별
Screening of salt-tolerance plants using transgenic Arabidopsis that express a salt cress cDNA library
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
Salt cress (Thellungiella halophila or Thellungiellaparvula), species closely related to Arabidopsis thaliana,represents an extremophile adapted to harsh saline environments. To isolate salt-tolerance genes from this species, we constructeda cDNA library from roots and leaves of salt cress plantstreated with 200 mM NaCl. This cDNA library was subsequentlyshuttled into the destination binary vector [drivenby the cauliflower mosaic virus (CaMV) 35S promoter]designed for plant transformation and expression via recombination-assisted cloning. In total, 305,400 pools of transgenicBASTA-resistant lines were generated in Arabidopsis usingeither T. halophila or T. parvula cDNA libraries. These wereused for functional screening of genes involved in salttolerance. Among these pools, 168,500 pools were used forprimary screening to date from which 7,157 lines showedapparent salt tolerant-phenotypes in the initial screen. Asecondary screen has now identified 165 salt tolerant transgeniclines using 1,551 (10.6%) lines that emerged in the firstscreen. The prevalent phenotype in these lines includes acceleratedseed germination often accompanied by faster root growthcompared to WT Arabidopsis under salt stress condition. Inaddition, other lines showed non-typical development of stemsand flowers compared to WT Arabidopsis. Based on the closerelationship of the tolerant species to the target species wesuggest this approach as an appropriate method for the largescaleidentification of salt tolerance genes from salt cress.
Abstract
Salt cress (Thellungiella halophila or Thellungiellaparvula), species closely related to Arabidopsis thaliana,represents an extremophile adapted to harsh saline environments. To isolate salt-tolerance genes from this species, we constructeda cDNA library from roots and leaves of salt cress plantstreated with 200 mM NaCl. This cDNA library was subsequentlyshuttled into the destination binary vector [drivenby the cauliflower mosaic virus (CaMV) 35S promoter]designed for plant transformation and expression via recombination-assisted cloning. In total, 305,400 pools of transgenicBASTA-resistant lines were generated in Arabidopsis usingeither T. halophila or T. parvula cDNA libraries. These wereused for functional screening of genes involved in salttolerance. Among these pools, 168,500 pools were used forprimary screening to date from which 7,157 lines showedapparent salt tolerant-phenotypes in the initial screen. Asecondary screen has now identified 165 salt tolerant transgeniclines using 1,551 (10.6%) lines that emerged in the firstscreen. The prevalent phenotype in these lines includes acceleratedseed germination often accompanied by faster root growthcompared to WT Arabidopsis under salt stress condition. Inaddition, other lines showed non-typical development of stemsand flowers compared to WT Arabidopsis. Based on the closerelationship of the tolerant species to the target species wesuggest this approach as an appropriate method for the largescaleidentification of salt tolerance genes from salt cress.
참고문헌 (25)
- Bent AF, "Arabidopsis in planta transformation. Uses, mechanisms, and prospects for transformation of other species", Plant Physiol, 124, pp.1540~1547 (2000)
- Bliss RD, "Osmotic sensitivity in relation to salt sensitivity in germinating barley seeds", Plant, Cell and Environment, 9, pp.721~725 (1986)
- Clough SJ, "Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana", Plant J, 16, pp.735~743 (1998)
- Cushman JC, "Genomic approaches to plant stress tolerance", Curr Opin Plant Biol, 3, pp.117~124 (2000)
- Dassanayake M, "The genome of extremophile crucifer Thellungiella parvula", Nature Genetics, 43, pp.913~918 (2013)
- Drews GN, "Negative regulation of the Arabidopsis homeotic gene AGAMOUS by the APETALA2 product", Cell, 65, pp.991~1002 (1991)
- Du J, "Functional gene-mining for salttolerance genes with the power of Arabidopsis", Plant J, 56, pp.653~664 (2008)
- Finkelstein RR, "Abscisic acid signaling in seeds and seedlings", Plant Cell(Suppl), 14, pp.S15~S45 (2002)
- Gong Q, "Salinity stress adaptation competence in the extremophile Thellungiella halophila in comparison with its relative Arabidopsis thaliana", Plant J, 44, pp.826~839 (2005)
- Inan G, "Salt Cress. A Halophyte and Cryophyte Arabidopsis Relative Model System and Its Applicability to Molecular Genetic Analyses of Growth and Development of Extremophiles", Plant Physiol, 135, pp.1718~1737 (2004)
- Kim WY, "Release of SOS2kinase from sequestration with GIGANTEA determines salt tolerance in Arabidopsis", Nature Communications, 4, pp.1352~ (2013)
- Krysan PJ, "T-DNA as an Insertional mutagen in Arabidopsis", Plant Cell, 11, pp.2283~2290 (1999)
- Lei Z-Y, "High-throughpur binary vectors for plant gene function analysis", J Integr Plant Biol, 49, pp.556~567 (2007)
- Munns R, "Mechanisms of salinity tolerance", Annual Review of Plant Biology, 59, pp.651~681 (2008)
- Ni WS, "Construction of a plant transformation-ready expression cDNA library for Thellungiella halophila using recombination cloning", J. Integr. Plant Biol, 49, pp.1313~1319 (2007)
- Papdi C, "Functional identification of Arabidopsis stress regulatory genes using the controlled cDNA overexpression system", Plant Physiol, 147, pp.528~542 (2008)
- Parvaiz A, "Salt stress and phyto-biochemical responses of plants –a review", Plant Soil Environ, 54, pp.88~99 (2008)
- Vallejo AJ, "Germination variation in Arabidopsis thaliana accessions under moderate osmotic and salt stresses", Ann of Botany, pp.1~10 (2010)
- Wu HJ, "Insights into salt tolerance from the genome of Thellungiella salsuginea", Proc Natl Acad Sci USA, 109, pp.12219~12224 (2012)
- Yamaguchi-Shinozaki K, "Transcriptional regulatory newtorks in cellular responses and tolerance to dehydration and cold stresses", Annu Rev Plant Biol, 53, pp.781~803 (2006)
- Yanofsky MF, "The protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors", Nature, 346, pp.35~39 (1990)
- 윤대진, "식물의 고염 스트레스에 대한 반응 및 적응기작(총설)", Journal of Plant Biotechnology, 32(1), pp.1~14 (2005), 한국식물생명공학회
- Zhu J-K, "Genetic analysis of salt tolerance in Arabidopsis: Evidence for a critical role of Potassium nutrition", Plant Cell, 10, pp.1181~1191 (1998)
- Zhu J-K, "Plant salt tolerance", Trends Plant Sci, 6, pp.66~71 (2001)
- Zhu J-K, "Salt and drought stress signal transduction in plants", Annu Rev Plant Biol, 53, pp.247~273 (2002)