A novel method for high-frequency transgenic shoot regeneration via Agrobacterium tumefaciens in flax (Linum usitatissimum L.)
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In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having ‘pBIN 19’ plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with 500 μl of bacterial solution after drying in sterile cabin for 35 min were used.
Abstract
In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having ‘pBIN 19’ plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with 500 μl of bacterial solution after drying in sterile cabin for 35 min were used.
참고문헌 (36)
- Amoah BK, "Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue", J Exp Bot, 52, pp.1135~1142 (2001)
- Beranova M, "Sonication assisted Agrobacterium-mediated transformation enhances the transformation efficiency in flax (Linum usitatissimum L.)", Plant Cell Tiss Organ Cult, 94, pp.253~259 (2008)
- Bevan M, "Multiple transcripts of T-DNA detected in nopaline crown gall tumors", J Mol Appl Genet, 1, pp.539~ (1982)
- Binns AN, "Cell biology of Agrobacterium infection and transformation of plants", Annu Rev Microbiol, 42, pp.575~606 (1988)
- Caillot S, "Influence of light intensity and selection scheme on regeneration time of transgenic flax plants", Plant Cell Rep, 28, pp.359~371 (2009)
- Chakrabarty R, "Agrobacterium-mediated transformation of cauliflower: optimization of protocol and development of Bt-transgenic cauliflower", J Biosci, 27, pp.495~502 (2002)
- De Clercq J, "An optimized Agrobacterium-mediated transformation procedure for Phaseolus acutifolius A. Gray", Plant Cell Rep, 21, pp.333~334 (2002)
- Dong J, "An improved procedure for production of transgenic flax plants using Agrobacterium tumefaciens", Plant Science, 88, pp.61~71 (1993)
- Friedt W, "Biotechnology in breeding of industrial oil crops:the present status and future prospects", Fat Sci Technol, 90, pp.51~55 (1990)
- Gamborg OL, "Tissue culture, protoplasts, and morphogenesis in flax", Bot Gaz, 137, pp.301~306 (1976)
- Green AG, "Isolation of induced mutants in linseed (Linum usitatissimum) having reduced linolenic acid content", Euphytica, 33, pp.321~328 (1984)
- Haq IU, "Agrobacterium-mediated transformation of cotton (Gossypium hirsutum L.) via vacuum infiltration", Plant Mol Biol Rep, 22, pp.279~288 (2004)
- Henzi MX, "Factors that influence Agrobacterium rhizogenesmediated transformation of broccoli (Brassica oleracea L. var. italica)", Plant Cell Rep, 19, pp.994~999 (2000)
- Hooykaas PJJ, "Agrobacterium and plant genetic engineering", Plant Mol Biol, 19, pp.15~38 (1992)
- Jefferson RA, "Assaying chimeric genes in plants: the GUS gene fusion system", Plant Mol Biol Rep, 5, pp.387~405 (1987)
- Jordan MC, "Transformed calllus does not necessarily regenerate transformed shoots", Plant Cell Rep, 7, pp.285~287 (1988)
- Joubert P, "Effects of phenolic compounds on Agrobacterium vir genes and gene transfer induction. A plausible molecular mechanism of phenol binding protein activation", Plant Sci, 162, pp.733~743 (2002)
- Le VQ, "An improved procedure for production of white spruce (Picea glauca) transgenic plants using Agrobacterium tumefaciens", J Exp Bot, 52, pp.2089~2095 (2001)
- Leelavathi S, "A simple and rapid Agrobacteriummediated transformation protocol for cotton (Gossypium hirsutum L.): Embryogenic calli as a source to generate large numbers of transgenic plants", Plant Cell Rep, 22, pp.465~470 (2004)
- Lopez SJ, "Agrobacterium tumefaciensmediated genetic transformation in tea (Camellia sinensis〔L.〕O. Kuntze)", Plant Mol Biol Rep, 22, pp.201a~201j (2004)
- Lorenc-Kukula K, "Pleiotropic Effect of Phenolic Compounds Content Increases in Transgenic Flax Plant", J. Agric. Food Chem, 53, pp.3685~3692 (2005)
- Mahmoudian M, "Transformation of lentil (Lens culinaris M.) cotyledonary nodes by vacuum infiltration of Agrobacterium tumefaciens", Plant Mol Biol Rep, 20, pp.251~257 (2002)
- Mathews VH, "Phytohormone control of regeneration in cultured tissues of flax", Z. Pflanzenphysiol, 80, pp.436~442 (1976)
- Millam S, "The use of flax (Linum usitatissimum) as a model system for studies on organogenesis in vitro: the effect of different carbonhydrates", Plant Cell Tiss Org, 28, pp.163~166 (1992)
- Mlynarova L, "High efficiency Agrobacterium-mediated gene traansfer to flax", Plant Cell Rep, 13, pp.282~285 (1994)
- O. Kuntze cv. Kangra Jat) plants obtained by Agrobacterium-mediated transformation of somatic embryos, "Plant Cell Rep", 2001, 712~720
- Murashige T, "A revised medium for rapid growth and bioassays with tobacco tissue culture", Physiol Plant, 15, pp.473~479 (1962)
- Snedecor GW, "Statistical Methods" (1967), Iowa State University Press
- Somleva MN, "Agrobacteriummediated transformation of switchgrass", Crop Sci, 42, pp.2080~2087 (2002)
- Spokevicius AV, "Agrobacterium-mediated in vitro transformation of wood-producing stem segments in eucalypts", Plant Cell Rep, 23, pp.617~624 (2005)
- Wang WC, "Development of a novel Agrobacterium-mediated transformation method to recover transgenic Brassica napus plants", Plant Cell Rep, 22, pp.274~281 (2003)
- Yildiz M, "The effect of sodium hypochlorite solutions on in vitro seedling growth and shoot regeneration of flax (Linum usitatissimum)", Naturwissenschaften, 89, pp.259~261 (2002)
- Yıldız, "Increasing the injured area on hypocotyl explants of flax (Linum usitatissimum L.) leads to high frequency callus-based shoot regeneration", Turk J Biol, 26, pp.95~98 (2002)
- Yıldız M, "The effect of different medium, gelling agents and explant ages on shoot regeneration from hypocotyl explants of flax (Linum usitatissimum L.)", 13th Biotechnology Congress, pp.137~144 (2003)
- Yildiz M, "The effect of a submersion pretreatment on in vitro explant growth and shoot regeneration from hypocotyls of flax (Linum usitatissimum)", Plant Cell Tiss Org, 77, pp.111~115 (2004)
- Zupan J, "The transfer of DNA from Agrobacterium tumefaciens into plants: a feast of fundamental insights", Plant J, 23, pp.11~28 (2000)