Abstract
The human Bcl-xL gene was transformed into peanut cultivar Georgia Green via microprojectile bombardment. Following selection on hygromycin-containing medium and regeneration, eighty hygromycin-resistant callus clusters were recovered. Southern blot analysis of ten fertile lines revealed multiple insertions of the Bcl-xL transgene in most lines. Western blot analysis of primary plants and T1 progenies demonstrated detectable levels of Bcl-xL expression in four transgenic lines. We could not detect Bcl-xL protein in other tested lines even though transcripts were identified by RT-PCR and northern blot. Three of the western-positive transgenic lines either were sterile or the progenies lost the expressive copy of Bcl-xL. Only T1 progenies from line BX25-4-2a-19 continued to express an intermediate level of Bcl-xL. This line demonstrated paraquat tolerance at the 5 μM level. Tolerance to salt of T1 and T2 seeds from seven other transgenic lines also was tested, but no tolerance was found in these lines. A high level of Bcl-xL transgene expression may be deleterious to plant growth and development even though the gene may confer tolerance to other abiotic and biotic stresses such as drought and pathogens.
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Awada T, Dunigan DD, Dickman MB (2003) Animal anti-apoptotic genes ameliorate the loss of turgor in water-stressed transgenic tobacco. Can J Plant Sci 83:499–506
Boise LH, Gonzalez-Garcia M, Postema CE, Ding L, Lindsten T, Turka LA, Mao X, Nunez G, Thompson CB (1993) bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell 74:597–608
Chen S, Dickman MB (2004) Bcl-2 family members localize to tobacco chloroplasts and inhibit programmed cell death induced by chloroplast-targeted herbicides. J Exp Bot 55:2617–2623
del Pozo O, Lam E (2003) Expression of the baculovirus p35 protein in tobacco affects cell death progression and compromises N gene-mediated disease resistance response to Tobacco mosaic virus. Mol Plant Microbe Interact 16:485–494
Dickman MB, Park YK, Oltersdorf T, Li W, Clemente T, French R (2001) Abrogation of disease development in plants expressing animal antiapoptotic genes. Proc Natl Acad Sci USA 98:6957–6962
Doukhanina EV, Chen S, van der Zalm E, Godzik A, Reed J, Dickman MB (2006) Identification and functional characterization of the BAG protein family in Arabidopsis thaliana. J Biol Chem 281:18793–18801
Gechev TS, van Breusegem F, Stone JM, Denev I, Laloi C (2006) Reactive oxygen species as signals that modulate plant stress responses and programmed cell death. Bioessays 28:1091–1101
Hansen G (2000) Evidence for Agrobacterium-induced apoptosis in maize cells. Mol Plant Microbe Interact 13:649–657
Hofius D, Tsitsigiannis DI, Jones JD, Mundy J (2007) Inducible cell death in plant immunity. Semin Cancer Biol 17:166–187. doi:10.1016/j.semcancer.2006.1012.1001
Kang CH, Jung WY, Kang YH, Kim JY, Kim DG, Jeong JC, Baek DW, Jin JB, Lee JY, Kim MO, Chung WS, Mengiste T, Koiwa H, Kwak SS, Bahk JD, Lee SY, Nam JS, Yun DJ, Cho MJ (2006) AtBAG6, a novel calmodulin-binding protein, induces programmed cell death in yeast and plants. Cell Death Differ 13:84–95
Kim R (2005) Unknotting the roles of Bcl-2 and Bcl-xL in cell death. Biochem Biophys Res Commun 333:336–343
Li W, Dickman MB (2004) Abiotic stress induces apoptotic-like features in tobacco that is inhibited by expression of human Bcl-2. Biotechnol Lett 26:87–95
Lincoln JE, Richael C, Overduin B, Smith K, Bostock R, Gilchrist DG (2002) Expression of the antiapoptotic baculovirus p35 gene in tomato blocks programmed cell death and provides broad-spectrum resistance to disease. Proc Natl Acad Sci U S A 99:15217–15221
Mitsuhara I, Malik KA, Miura M, Ohashi Y (1999) Animal cell-death suppressors Bcl-x(L) and Ced-9 inhibit cell death in tobacco plants. Curr Biol 9:775–778
Morino K, Olsen O, Shimamoto K (1999) Silencing of an aleurone-specific gene in transgenic rice is caused by a rearranged transgene. Plant J 17:275–285
Morino K, Olsen O, Shimamoto K (2004) Silencing of the aleurone-specific Ltp2-gus gene in transgenic rice is reversed by transgene rearrangements and loss of aberrant transcripts. Plant Cell Physiol 45:1500–1508
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497
Netondo GW, Onyango JC, Beck E (2004) Sorghum and salinity: II. Gas exchange and chlorophyll fluorescence of sorghum under salt stress. Crop Sci 44:806–811
Noji M, Saito M, Nakamura M, Aono M, Saji H, Saito K (2001) Cysteine synthase overexpression in tobacco confers tolerance to sulfur-containing environmental pollutants. Plant Physiol 126:973–980
Ozias-Akins P, Gill R (2001) Progress in the development of tissue culture and transformation methods applicable to the production of transgenic peanut. Peanut Sci 28:123–131
Ozias-Akins P, Schnall JA, Anderson WF, Singsit C, Clemente T, Adang MJ, Weissinger AK (1993) Regeneration of transgenic peanut plants from stably transformed embryogenic callus. Plant Sci 93:185–194
Polidoros AN, Mylona PV, Scandalios JG (2001) Transgenic tobacco plants expressing the maize Cat2 gene have altered catalase levels that affect plant-pathogen interactions and resistance to oxidative stress. Transgenic Res 10:555–569
Qiao J, Mitsuhara I, Yazaki Y, Sakano K, Gotoh Y, Miura M, Ohashi Y (2002) Enhanced resistance to salt, cold and wound stresses by overproduction of animal cell death suppressors Bcl-xL and Ced-9 in tobacco cells—their possible contribution through improved function of organella. Plant Cell Physiol 43:992–1005
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd ed. Cold Spring Harbor Laboratory Press, New York
Sharma AD, Gill PK, Singh P (2002) DNA isolation from dry and fresh samples of polysaccharide-rich plants. Plant Mol Biol Rep 20:415a–415f
Singsit C, Adang MJ, Lynch RE, Anderson WF, Wang A, Cardineau G, Ozias-Akins P (1997) Expression of a Bacillus thuringiensis cryIA(c) gene in transgenic peanut plants and its efficacy against lesser cornstalk borer. Transgenic Res 6:169–176
Topfer R, Maas C, Horicke-Grandpierre C, Schell J, Steinbiss H-H (1993) Expression vectors for high-level gene expression in dicotyledonous and monocotyledonous plants. Meth Enzymol 217:66–78
Watanabe N, Lam E (2004) Recent advance in the study of caspase-like proteases and Bax-inhibitor-1 in plants: their possible roles as regulator of programmed cell death. Mol Plant Path 5:65–70
Xu P, Rogers SJ, Roossinck MJ (2004) Expression of antiapoptotic genes bcl-xL and ced-9 in tomato enhances tolerance to viral-induced necrosis and abiotic stress. Proc Natl Acad Sci U S A 101:15805–15810
Yang H, Nairn J, Ozias-Akins P (2003) Transformation of peanut using a modified bacterial mercuric ion reductase gene driven by an actin promoter from Arabidopsis thaliana. J Plant Physiol 160:945–952
Acknowledgments
Support for this work was provided by the USDA Multicrop Aflatoxin Elimination Program and the National Peanut Foundation. We thank Evelyn P. Morgan for her technical assistance and Benjamin G. Mullinix for his help on statistical analysis. We also thank Marty Dickman, Texas A & M University and IDUN Pharmaceuticals (now part of Pfizer) for providing the Bcl-xL gene.
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Communicated by K. Kamo.
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Chu, Y., Deng, X.Y., Faustinelli, P. et al. Bcl-xL transformed peanut (Arachis hypogaea L.) exhibits paraquat tolerance. Plant Cell Rep 27, 85–92 (2008). https://doi.org/10.1007/s00299-007-0444-2
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DOI: https://doi.org/10.1007/s00299-007-0444-2