In vivo evaluation of transgenic watercress containing gene encoding Escherichia coli heat-labile toxin B subunit
Tóm tắt
In this work, the characterizations of the LTB (Escherichia coli heat-labile toxin B subunit) transgenic watercresses through Agrobacterium tumefaciens-mediated transformation (A1 and A3-A5) and by using biolistic method (B1 and B4) were investigated. Generally, their growth is not remarkably different from the wild-type. Their physiological and biochemical characteristics are relatively different in which plant A1 has highest values such as pigment (0.92 mg g−1), photosynthetic rate (23.39 mgCO2 [dm2]−1 h−1), dry matter (7.42 %), vitamin C (0.34 mg g−1), calcium (0.83 mg g−1), and potassium (2.47 mg g−1). The dry matter and calcium of all the transgenic watercresses and the wild-type are the same content. Southern blot hybridization showed the transgenic plants contain 1–2 copies in the genome. LTB protein strongly expresses in all the transgenic plants with contents from 1.16 to 1.46 % of total soluble protein. The GM1-ELISA binding assay indicated that plant-derived LTB protein bound to GM1-ganglioside receptors.
Tài liệu tham khảo
Arakawa T, Chong DK, Merritt JL, Langridge WH (1997) Expression of cholera toxin B subunit oligomers in transgenic potato plants. Transgenic Res 6:403–413
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Chen Y, Xu DQ (2007) Changes in leaf photosynthesis of transgenic rice with silenced OsBP-73 gene. Photosynthetica 45:419–425
Chevallier A (1996) The encyclopedia of medicinal plants. Dorling Kindersley, London
Chikwamba R, McMurray J, Shou H, Frame B, Pegg SE, Scott P, Manson H, Wang K (2002) Expression of a synthetic E.coli heat-labile enterotoxin B sub-unit (LT-B) in maize. Mol Breed 10:253–265
Grodzinxki AM, Grodzinxki DM (1981) Handbook of Plant Physiology. Technology and Science Publishing House, Ha Noi (in Vietnamese)
Jin RG, Liu YB, Tabashnik BE, Borthakur D (1999) Tissue culture and Agrobacterium-mediated transformation of watercress. Plant Cell Tissue Organ Cult 58:171–176
Kang TJ, Loc NH, Jang MO, Jang YS, Kim YS, Seo JE, Yang MS (2003) Expression of the B subunit of E.coli heat-labile enterotoxin in the chloroplasts of plants and its characterization. Transgenic Res 12:683–691
Kang TJ, Han SC, Jang MO, Kang KH, Jang YS, Yang MS (2004) Enhanced expression of B-subunit of Escherichia coli heat-labile enterotoxin in tobacco by optimization of coding sequence. Appl Biochem Biotechnol 117:175–87
Khoa LV (2000) Methods of analysis of soil, water, fertilizer and crop. Education Publishing House, Ha Noi (in Vietnamese)
Kiełkiewicz M, Gajc-Wolska J, Ślusarz S, Szwacka M (2012) Growth, development and yield of transgenic 35S-thaumatin II-expressing cucumber plants – open field evaluation. Sci Hortic 143:82–91
Li YX, Chang FQ, Du LQ, Guo BH, Li HJ, Zhang JS, Chen SY, Zhu ZQ (2000) Genetic transformation of watercress with a gene encoding for betaine-aldehyde dehydrogenase (BADH). J Integr Plant Biol 42:315–318
Lieman-Hurwitz J, Rachmilevitch S, Mittler R, Marcus Y, Kaplan A (2003) Enhanced photosynthesis and growth of transgenic plants that express ictB, a gene involved in HCO3 − accumulation in cyanobacteria. Plant Biotechnol J 1:43–50
Loc NH, Bach NH, Kim TG, Yang MS (2010a) Tissue culture and expression of Escherichia coli heat-labile enterotoxin B subunit in transgenic Peperomia pellucida. Protein Expr Purif 72:82–86
Loc NH, Song NV, Tien NQD, Minh TT, Nga PTQ, Phuong TTB, Kim TG, Yang MS (2010b) Expression of heat-labile enterotoxin B subunit from E. coli in transgenic watercress. Vietnam J Biotechnol 8:1173–1179
Loc NH, Song NV, Tien NQD, Minh TT, Nga PTQ, Phuong TTB, Kim TG, Yang MS (2011) Expression of the Escherichia coli heat-labile enterotoxin B subunit in transgenic watercress (Nasturtium officinale L.). Plant Cell Tissue Organ Cult 105:39–45
Loc NH, Long DT, Nhi PY, Hong HTK, Kim TG, Yang MS (2012) Preliminary evaluation of transgenic tomato plants expressing Escherichia coli heat-labile toxin B subunit grown under in vivo condition. Ann Biol Res 3:2070–2073
Mui NV (2001) Practice in Biochemistry. Technology and Science Publishing House, Ha Noi (in Vietnamese)
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Nhi PY, Kim TG, Yang MS, Loc NH (2010) Expression of Escherichia coli heat-labile enterotoxin B subunit in transgenic tomato (Lycopersicon esculentum L.). Vietnam J Biotechnol 8:1279–1285
Ogita S, Usui M, Shibutani N, Kato Y (2009) A simple shoot multiplication procedure using internode explants, and its application for particle bombardment and Agrobacterium-mediated transformation in watercress. J Plant Res 122:455–463
Phillips R, Rix M (1995) Vegetables. Macmillan Reference Books, London
Reinecke DM, Wickramarathna AD, Ozga JA, Kurepin LV, Jin AL, Good AG, Pharis RP (2013) Gibberellin 3-oxidase gene expression patterns influence gibberellin biosynthesis, growth, and development in pea. Plant Physiol 163(2):929–45. doi:10.1104/pp.113.225987
Rezaee MA, Rezaee A, Moazzeni SM, Salmanian AH, Yasuda Y, Tochikubo K, Pirayeh SN, Arzanlou M (2005) Expression of Escherichia coli heat-labile enterotoxin B subunit (LTB) in Saccharomyces cerevisiae. J Microbiol 43:354–360
Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory, Plainview
Thinh LT, Long DT (2011) Morphological and qualitative characteristics of CTB (cholera toxin B subunit) transgenic tomato fruit. J Sci Hue Univ 64:159–165
Tochikubo K, Yasuda Y (2000) Principles of mucosal immunity and development of mucosal vaccines using cholera toxin B subunit and its related adjuvants. Recent Res Dev Microbiol 4:387–405
Wang GK, Zhang M, Gong JF, Guo QF, Feng YN, Wang W (2012) Increased gibberellin contents contribute to accelerated growth and development of transgenic tobacco overexpressing a wheat ubiquitin gene. Plant Cell Rep 31:2215–27
Yao Q, Cong L, Chang JL, Li KX, Yang GX, He GY (2006) Low copy number gene transfer and stable expression in a commercial wheat cultivar via particle bombardment. J Exp Bot 57:3737–3746