Abstract
Purple-colored transgenic creeping bentgrass (Agrostis stolonifera L.) plants were developed for ornamental purpose by means of Agrobacterium-mediated transformation. Embryogenic creeping bentgrass calli were transformed with the pCAMBIA 3301 vector harboring maize (Zea mays) flavonoid/anthocyanin biosynthetic pathway transcription factor genes, Lc (Leaf color) and Pl (Purple leaf), individually and in combination, and three types of putative transgenic plants (Lc, Pl, and Lc + Pl) were generated. Genomic integration and expression of the transgenes were confirmed by Southern and northern blot analyses, respectively. The transgenic creeping bentgrass plants expressing both Lc and Pl genes were entirely purple, whereas those expressing Pl alone had purple stems and those expressing Lc alone lacked purple pigmentation in adult plants. The anthocyanin content was estimated in all the three types of transgenic plant and correlated well with the degree of purple coloration observed. These results suggest that both Lc and Pl genes are necessary and sufficient to confer purple coloration to creeping bentgrass.
Similar content being viewed by others
References
Boase MR, Bradley JM, Borst NK (1998) Genetic transformation mediated by Agrobacterium tumefaciens of florists’ chrysanthemum (Dendranthema × Grandiflorum) cultivar “Peach Margaret”. In Vitro Cell Dev Biol 34:46–51
Bonos SA, Plumley KA, Meyer WA (2002) Ploidy determination in Agrotis using flow cytometry and morphological traits. Crop Sci 42:192–196
Bovy A, de Vos R, Kemper M, Schijlen E, Pertejo MA, Muir S, Collins G, Robinson S, Verhoeyen M, Hughes S, Santos-Buelga C, van Tunen A (2002) High-Flavonol tomatoes resulting from the heterologous expression of the maize transcription factor genes LC and C1. Plant Cell 14:2509–2526
Bradley JM, Davies KM, Deroles SC, Bloor SJ, Lewis DH (1998) The maize Lc regulatory gene up-regulates the flavonoid biosynthetic pathway of Petunia. Plant J 13:381–392
Chai B, Sticklen MB (1998) Applications of biotechnology in turfgrass genetic improvement. Crop Sci 38:1320–1338
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 Sci 4:346–351
Chen H, Nelson RS, Sherwood JL (1994) Enhanced recovery of transformants of Agrobacterium tumefaciens after freeze-thaw transformation and drug selection. Biotechniques 16:664–670
Cheng M, Lowe BA, Spencer TM, Ye X, Armstrong CL (2004) Factors influencing Agrobacterium-mediated transformation of monocotyledonous species. In Vitro Cell Dev Biol Plant 40:31–45
Cone KC, Cocciolone SM, Burr FA, Burr B (1993) Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant. Plant Cell 5:1795–1805
Dai S, Zheng P, Marmey P, Zhang S, Tian W, Chen S, Beachy RN, Fauquet C (2001) Comparative analysis of transgenic rice plants obtained by Agrobacterium-mediated transformation and particle bombardment. Mol Breed 7:25–33
Dooner HK, Robbins TP, Jorgensen RA (1991) Genetic and developmental control of anthocyanin biosynthesis. Annu Rev Genet 25:173–199
Fu D, Tisserat NA, Xiao Y, Settle D, Muthukrishnan S, Liang GH (2005) 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 barley hva1 gene in creeping bentgrass for improving drought resistance. Plant Cell Rep 26:467–477
Goldsbrough AP, Tong Y, Yoder JI, Tong Y (1996) Lc as a non-destructive visual reporter and transposition excision marker gene for tomato. Plant J 9:927–933
Harborne JB, Williams CA (2001) Anthocyanins and other flavonoids. Nat Prod Rep 18:310–333
Hartman CL, Lee L, Day RR, Tumer NE (1994) Herbicide resistant turfgrass (Agrostis palustris Huds.) by biolistic transformation. Bio/Technology 12:919–923
Holton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071–1083
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Kim SJ, Lee JY, Kim YM, Yang SS, Hwang OJ, Hong NJ, Kim KM, Lee HY, Song PS, Kim JI (2007) Agrobacterium-mediated high efficiency transformation of creeping bentgrass with herbicide resistance. J Plant Biol 50:577–585
Lee L (1996) Turfgrass biotechnology. Plant Sci 115:1–8
Li SJ, Deng XM, Mao HZ, Hong Y (2005) Enhanced anthocyanin synthesis in foliage plant Caladium bicolor. Plant Cell Rep 23:716–720
Li H, Flachowsky H, Fischer TC, Hanke MV, Forkmann G, Treutter D, Schwab W, Hoffmann T, Szankowski I (2007) Maize Lc transcription factor enhances biosynthesis of anthocyanins, distinct proanthocyanidins and phenylpropanoids in apple (Malus domestica Borkh.). Planta 226:1243–1254
Liang GH, Skinner DZ (2004) Genetically modified crops: their development, uses, and risks. Food Product Press, An Imprint of the Haworth Press Inc., New York
Lloyd AM, Walbot V, Davis RW (1992) Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1. Science 258:1773–1775
Ludwig SR, Wessler SR (1990) Maize R gene family: tissue-specific heleix-loop-helix proteins. Cell 62:849–851
Ludwig SR, Habera LF, Dellaporta ST, Wessler SR (1989) Lc, a member of the maze R gene family responsible for tissue-specific anthocyanin production, encodes a protein similar to transcriptional activators and contains the myc-homolgy region. Proc Natl Acad Sci USA 86:7092–7096
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 Rep 22:645–652
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Pilu R, Piazza P, Petroni K, Ronchi A, Martin C, Tonelli C (2003) pl-bol3, a complex allele of the anthocyanin regulatory pl1 locus that arose in a naturally occurring maize populations. Plant J 36:510–521
Quattrocchio F, Wing JF, Leppen HTC, Mol JNM, Koes R (1993) Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes. Plant Cell 5:1497–1512
Quattrocchio F, Wing JF, van der Woude K, Mol JNM, Koes R (1998) Analysis of bHLH and MYB domain proteins: species-specific regulatory differences are caused by divergent evolution of target anthocyanin genes. Plant J 13:475–488
Ray H, Yu M, Auser P, Blahut-Beatty L, McKersie B, Bowley S, Westcott N, Coulman B, Lloyd A, Gruber MY (2003) Expression of anthocyanins and proanthocyanidins after transformation of alfalfa with Maize Lc. Plant Physiol 132:1448–1463
Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York
Springob K, Nakajima J, Yamazaki M, Saito K (2003) Recent advances in the biosynthesis and accumulation of anthocyanins. Nat Prod Rep 20:288–303
Tanaka Y, Sasaki N, Ohmiya A (2008) Biosynthesis of plant pigments: anthocyanins, betalains and carotenoids. Plant J 54:733–749
Travella S, Ross SM, Harden J, Everett C, Snape JW, Harwood WA (2005) A comparison of transgenic barley lines produced particle bombardment and Agrobacterium mediated techniques. Plant Cell Rep 23:780–789
Wang Z, Hopkins A, Mian R (2001) Forage and turf grass biotechnology. Crit Rev Plant Sci 20:573–619
Warnke S (2003) Creeping bentgrass (Agrostis stolonifera L.). In: Casler MD, Duncan RR (eds) Turfgrass biology, genetics, and breeding. Wiley, New Jersey, pp 175–185
Winkel-Shirley B (2001) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126:485–493
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. Hereditas 133:229–233
Zhong H, Boyland MG, Srinivasan C, Sticklen MB (1993) Transgenic plants of turfgrass (Agrostis palustris Huds.) from microprojectile bombardment of embryogenic callus. Plant Cell Rep 13:1–6
Acknowledgments
We thank the Kumho Life Science Laboratory in Chonnam National University for providing plant growth facilities. This work was supported by the BioGreen 21 Program (Code # 20080401034014), Rural Development Administration, Republic of Korea, and the KOSEF/MOST to the Environmental Biotechnology National Core Research Center (NCRC) (Grant # R15-2003-012-01003-0).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by J. Register.
Rights and permissions
About this article
Cite this article
Han, YJ., Kim, YM., Lee, JY. et al. Production of purple-colored creeping bentgrass using maize transcription factor genes Pl and Lc through Agrobacterium-mediated transformation. Plant Cell Rep 28, 397–406 (2009). https://doi.org/10.1007/s00299-008-0648-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-008-0648-0