Agrobacterium-medlated high-efficiency transformation of creeping bentgrass with herbicide resistance

Journal of Plant Biology - Tập 50 - Trang 577-585
Yong-Min Kim1, Nam-Ju Hong1, Ok-Jin Hwang1, Song-Sook Yang1, Jeong-II Kim1,2, Pill-Soon Song3,2, Hyo-Yeon Lee3, Jee-Yeon Lee1, Kyung-Moon Kim3, Soo Jung Kim1
1Department of Molecular Biotechnology (BK21 Program) and Kumho Life Science Laboratory, Chonnam National University, Gwangju, Korea
2Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju, Korea
3Chcju National University, Jeju, Korea

Tóm tắt

We performedAgrobacterium-mediated genetic transformation of creeping bentgrass(Agrostis stolonifera L.) and produced herbicide-resistant transformants from commercial cultivars Crenshaw and Penncross. Seed-derived embryogenie calli were infected withA. tumefaciens EHA105 harboring pCAMBIA 3301, which includes an intron-containinggus reporter and abar selection marker. To establish a stable system, we examined various factors that could potentially influence transformation efficiency during the pre-culture, infection, and co-cultivation steps. The addition of kinetin to the callus pre-culture media increased efficiency about three-fold. Once the optimum infection and co-cultivation conditions were identified, this protocol was used successfully to bulk-produce herbicide-resistant transgenic plants whose herbicide resistance was confirmed using the BASTA® resistance test. Southern blot analysis demonstrated integration and low copy numbers of the integrated transgenes, and northern blot analysis verified their expression. Thus, we have established an efficient genetic transformation system for creeping bentgrass and confirmed a high frequency of single-copy transgene integration and functional gene expression.

Tài liệu tham khảo

Asano Y, llo Y, Fukami M, Morifuji M, Fujiie A (1997) Production of herbicide resistant transgenic creeping bent plants. Intl Turfgrass Soc Res J8: 261–267 Bonos SA, Plumley KA, Meyer WA (2002) Ploidy determination inAgrostis using flow cytometry and morphological traits. Crop Sci42: 192–196 Chai B, Slicklen MB (1998) Applications of biotechnology in turfgrass genetic improvement. Crop Sci38: 1320–1338 Chai ML, Wang BL, Kim JY, Lee JM, Kim DH (2003) Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass. J Zhejiang Univ Sci4: 346–351 Chai ML, Senlhil KK, Kim DH (2004) Transgenic plants of colonial bentgrass from embryogenie callus viaAgrobacterium-mediated transformation. Plant Cell Tiss Org Cult77: 165–171 Chen H, Nelson RS, Sherwood JL (1994) Enhanced recovery of transformants ofAgrobacterium tumefaciens after freeze-thaw transformation and drug selection. Biotechniques16: 664–670 Chung KM, Park YD (2005) Development of an Agrobacterium-mediated transformation system for regenerating garland Chrysanthemum(Chrysanthemum coronarium L.). J Plant Biol48: 136–141 Dai S, Zheng P, Marmey R Zhang S, Tian W, Chen S, Beachy RN, Fauquel C (2001) Comparative analysis of transgenic rice plants obtained byAgrobacterium-mediated transformation and particle bombardment. Mol Breed7: 25–33 Dong S, Qu R (2005) High efficiency transformation of tall fescue withAgrobacterium tumefaciens. Plant Sci168: 1453–1458 Fu D, Xiao Y, Mulhukrishnan S, Liang GH (2005a) In vivo performance of a dual genetic marker,manA-gfp, in transgenic bentgrass. Genome48: 722–730 Fu D, Tisserat NA, Xiao Y, Settle D, Muthukrishnan S, Liang GH (2005b) Overexpression of rice TLPD34 enhances dollar-spot resistance in transgenic bentgrass. Plant Sci.168: 671–680 Fu D, Huang B, Xiao Y, Muthukrishnan S, Liang GH (2007) Overexpression of barleyhva1 gene in creeping bentgrass for improving drought resistance. Plant Cell Rep26: 467–477 Ge Y, Norton T, Wang ZY (2006) Transgenic zoysia grass(Zoysia japonica) plants obtainedby Agrobacterium-mediated transformation. Plant Cell Rep25: 792–798 Ge Y, Cheng X, Hopkins A, Wang ZY (2007) Generation of transgenicLolium temulentum plants byAgrobacterium tumefaciens-mediated transformation. Plant Cell Rep26: 783–789 Han N, Chen D, Bian HW, Deng MJ, Zhu MY (2005) Production of transgenic creeping bentgrassAgrostis stolonifera var.palustris plantsby Agrobacterium tumefaciens-mediated transformation using hygromycin selection. Plant Cell Tiss Org Cult81: 131–138 Hartman CL, Lee L, Day RR, Turner NE (1994) Herbicide resistant turfgrass (Agrostis palustris Huds.) by biolistic transformation. Bio/Technology12: 919–923 Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep5: 387–405 Kim DH, Chai ML, Senthil K, Lee JM, Park MH, Kim JY (2001) Factors affecting the transformation of bentgrass (Agrostis spp.) based onAgrobacterium tumefaciens. J Kor Soc Hort Sci43: 243–248 Lee L (1996) Turfgrass biotechnology. Plant Sci115: 1–8 Lohar DP, Schuller K, Buzas DM, Gresshoff PM, Stiller J, (2001) Transformation ofLotus japonicus using the herbicide resistant bar gene as a selectable marker. J Exp Bot52: 1 697–1702 Luo H, Hu Q, Nelson K, Longo C, Kausch AP, Chandlee JM, Wipff JK, Fricker CR (2004)Agrobacterium tumefaciens-mediated creeping bentgrass (Agrostis stolonifera L.) transformation using phosphinothricin selection results in a high frequency of single-copy transgene integration. Plant Cell Rep22: 645–652 Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant15: 473–497 Prince JP, Pochard E, Tanksley SD (1993) Construction of a molecular linkage map of pepper and a comparison of synteny with tomato. Genome36: 404–417 Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press, New York Toyama K, Bae CH, Kang JG, Lim YR, Adachi T, Riu KZ, Song PS, Lee HY (2003) Production of herbicide-tolerant zoysia grass byAgrobacterium-mediated transformation. Mol Cells16: 19–27 Wang ZY, Ge Y (2005)Agrobacterium-mediated high efficiency transformation of tall fescue (Festuca arundinacca). J Plant Physiol162: 103–113 Wang ZY, Ge Y (2006) Recent advances in genetic transformation of forage and turf grasses. In Vitro Cell Dev Biol Plant42: 1–18 Wang Z, Hopkins A, Mian R (2001) Forage and turf grass biotechnology. Crit Rev Plant Sci20: 573–619 Wang Y, Browning M, Ruemmele BA, Chandlee JM, Kausch AP (2003) Glufosinate reduces fungal diseases in transgenic glufosinate-resistant bentgrasses (Agrostis spp.) Weed Sci51: 130–137 Warnke S (2003) Creeping bentgrass (Agrostis stoloniiera L.),In MD Casier, RR Duncan, eds, Turfgrass Biology, Genetics, and Breeding. John Wiley & Sons, Inc., NJ, USA, pp 175–185 Wehrmann A, Vilel AD, Opsomer C, Bollerman J, Schulz A (1996) The similarities ofbar andpat gene products make them equally applicable for plant engineers. Nature Biotechnol14: 1274–1278 Yu TT, Skinner DZ, Liang GH, Trick HN, Huang B, Muthukrishnan S (2000)Agrobacterium-mediated transformation of creeping bentgrass using GFP as a reporter gene. Hereditas133: 229–233 Zhong H, Boyland MG, Srinivasan C, Slicklen MB (1993) Trans-genic plants of turfgrass (Agrostis palustris Huds.) from microprojectile bombardment of embryogenic callus. Plant Cell Rep13: 1–6 Zilinskas BA, Wang X (2004) Genetic transformation of turfgrass,In GH Liang, DZ Skinner, eds, Genetically Modified Crops: Their Development, Uses, and Risks. Food Product Press, an imprint of the Haworth Press, Inc., New York, pp 309–350