Isolation and characterization of an ubiquitin extension protein gene (JcUEP) promoter from Jatropha curcas
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The JcUEP promoter is active constitutively in the bio-fuel plant Jatropha curcas , and is an alternative to the widely used CaMV35S promoter for driving constitutive overexpression of transgenes in Jatropha.
Well-characterized promoters are required for transgenic breeding of Jatropha curcas, a biofuel feedstock with great potential for production of bio-diesel and bio-jet fuel. In this study, an ubiquitin extension protein gene from Jatropha, designated JcUEP, was identified to be ubiquitously expressed. Thus, we isolated a 1.2 kb fragment of the 5′ flanking region of JcUEP and evaluated its activity as a constitutive promoter in Arabidopsis and Jatropha using the β-glucuronidase (GUS) reporter gene. As expected, histochemical GUS assay showed that the JcUEP promoter was active in all Arabidopsis and Jatropha tissues tested. We also compared the activity of the JcUEP promoter with that of the cauliflower mosaic virus 35S (CaMV35S) promoter, a well-characterized constitutive promoter conferring strong transgene expression in dicot species, in various tissues of Jatropha. In a fluorometric GUS assay, the two promoters showed similar activities in stems, mature leaves and female flowers; while the CaMV35S promoter was more effective than the JcUEP promoter in other tissues, especially young leaves and inflorescences. In addition, the JcUEP promoter retained its activity under stress conditions in low temperature, high salt, dehydration and exogenous ABA treatments. These results suggest that the plant-derived JcUEP promoter could be an alternative to the CaMV35S promoter for driving constitutive overexpression of transgenes in Jatropha and other plants.
KeywordsPhysic nut Constitutive promoter Ubiquitin Transgenic Stress CaMV35S
This work was supported by funding from the Top Science and Technology Talents Scheme of Yunnan Province (2009CI123), the Natural Science Foundation of Yunnan Province (2011FA034) and the CAS 135 program (XTBG-T02) awarded to Z.-F. Xu. The authors gratefully acknowledge the Central Laboratory of the Xishuangbanna Tropical Botanical Garden for providing research facilities.
- de Argollo Marques D, Siqueira WJ, Colombo CA, Ferrari RA (2013) Breeding and biotechnology of Jatropha curcas. In: Bahadur B, Sujatha M, Carels N (eds) Jatropha, challenges for a New Energy Crop, vol 2., Genetic improvement and biotechnologySpringer, New York, pp 457–478Google Scholar
- Pan J, Fu Q, Xu ZF (2010) Agrobacterium tumefaciens-mediated transformation of biofuel plant Jatropha curcas using kanamycin selection. Afr J Biotechnol 9:6477–6481Google Scholar
- Park HC, Kim ML, Kang YH, Jeon JM, Yoo JH, Kim MC, Park CY, Jeong JC, Moon BC, Lee JH (2004) Pathogen-and NaCl-induced expression of the SCaM-4 promoter is mediated in part by a GT-1 box that interacts with a GT-1-like transcription factor. Plant Physiol 135:2150–2161CrossRefPubMedCentralPubMedGoogle Scholar
- Pérez-Barranco G, Torreblanca R, Padilla IMG, Sánchez-Romero C, Pliego-Alfaro F, Mercado JA (2009) Studies on genetic transformation of olive (Olea europaea L.) somatic embryos: I. Evaluation of different aminoglycoside antibiotics for nptII selection; II. Transient transformation via particle bombardment. Plant Cell Tiss Org 97:243–251CrossRefGoogle Scholar
- Sambrook J, Russell DW, Russell DW (2001) Molecular cloning: a laboratory manual (3-volume set). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
- Sato S, Hirakawa H, Isobe S, Fukai E, Watanabe A, Kato M, Kawashima K, Minami C, Muraki A, Nakazaki N, Takahashi C, Nakayama S, Kishida Y, Kohara M, Yamada M, Tsuruoka H, Sasamoto S, Tabata S, Aizu T, Toyoda A, Shin-i T, Minakuchi Y, Kohara Y, Fujiyama A, Tsuchimoto S, Kajiyama S, Makigano E, Ohmido N, Shibagaki N, Cartagena JA, Wada N, Kohinata T, Atefeh A, Yuasa S, Matsunaga S, Fukui K (2011) Sequence analysis of the genome of an oil-bearing tree, Jatropha curcas L. DNA Res 18:65–76CrossRefPubMedCentralPubMedGoogle Scholar
- Takimoto I, Christensen AH, Quail PH, Uchimiya H, Toki S (1994) Non-systemic expression of a stress-responsive maize polyubiquitin gene (Ubi-1) in transgenic rice plants. Plant Mol Biol 26:1007–1012Google Scholar