Browse > Article
http://dx.doi.org/10.5010/JPB.2010.37.1.072

Development of transgenic cucumber expressing TPSP gene and morphological alterations  

Kim, Hyun-A (Department of Medicinal Plant Resources, Nambu University)
Min, Sung-Ran (Korea Research Institute of Bioscience and Biotechnology)
Choi, Dong-Woog (Department of Biological Education, Chonnam National University)
Choi, Pil-Son (Department of Medicinal Plant Resources, Nambu University)
Hong, Seong-Gyun (Department of Medicinal Plant Resources, Nambu University)
Publication Information
Journal of Plant Biotechnology / v.37, no.1, 2010 , pp. 72-76 More about this Journal
Abstract
To develop transgenic cucumber tolerant to abiotic stress, a cotyledonary-node explants were co-cultivated with Agrobacterium tumefaciens (EHA101) carrying TPSP gene (pHC30-TPSP). After transfer to fresh medium every two week for eight weeks, putative transgenic plants were selected when shoots grown a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with $5\;mgl^{-1}$ phospinotricin as selectable agent. The confirmation of transgenic cucumber was based on the Northern blot analysis. Thirty four shoots (5.2%) with resistance to phospinotricin were obtained from 660 explants inoculated. Of them, transformants were only confirmed from 11 plants (1.7%). Transgenic cucumber expressing TPSP gene was more synthesized at 3.8 times amounts of trehalose (0.014 mg g fresh $wt^{-1}$) than non-transformants (0.0037 mg g fresh $wt^{-1}$). However, all of transgenic plants showed abnormal morphology, including stunted growth (< height 15 cm), shrunken leaves, and sterility as compared with non-transgenic plants (> height 150 cm) under the same growth environment. These results lead us to speculate that the overproduction of trehalose was toxic for cucumber, even though that had known for rice as non-toxic.
Keywords
Agrobacterium; stunted growth; transgenic cucumber; trehalose;
Citations & Related Records
Times Cited By KSCI : 4  (Citation Analysis)
연도 인용수 순위
1 Kim HA, Lee BY, Jeon JJ, Choi DW, Choi PS, Utomo SD, Lee JH, Kang TH, Lee YJ (2008) GUS gene expression and plant regeneration via somatic embryogenesis in cucumber (Cucumis sativus L.). Korean J Plant Biotech 35:275-280   과학기술학회마을   DOI   ScienceOn
2 Muller J, Wiemken A, Aeschbacher R (1999) Trehalose metabolism in sugar sensing and plant development. Plant Sci 147:37-47   DOI
3 Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473-497   DOI
4 Nishibayashi S, Kayakawa T, Nakajima T, Suzuki M, Kaneko H (1996) CMV protection in transgenic cucumber plants with an introduced CMV-O cp gene. Theor Appl Genet 93:672-678   DOI
5 Pilon-Smits EAH, Terry N, Sears T, Kim H, Zayed A, Hwang S, van Dun K, Voogd E, Verwoerd TC, Krutwagen RW (1998) Trehalose-producing transgenic tobacco plants show improved growth performance under drought stress. J Plant Physiol 152:525-532   DOI   ScienceOn
6 Romero C, Belles JM, Vaya JL, Serrano R, Culianez-Macia FA (1997) Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. Planta 201:293-297   DOI
7 Saramento GG, Alpert K, Tang FA, Punja ZK (1992) Factors influencing Agrobacterium tumefaciens mediated transformation and expression of kanamycin resistance in pickling cucumber. Plant Cell Tiss Organ Cult 31:185-193
8 Zhang Z, Xing A, Staswick P, Clemente TE (1999) The use of glufosinate as a selective agent in Agrobacterium-mediated transformation of soybean. Plant Cell Tiss Org Cult 56 :37-46   DOI
9 Eleutherio ECA, Araujo PS, Panek AD (1993) Protective role of trehalose during heat stress in Saccharomyces cerevisiae. Cryobiology 30:591-596   DOI
10 Gaba V, Zelcer A, Gal-On A (2004) Cucurbit biotechnology-the importance of virus resistance. In Vitro Cell Dev Biol-Plant 40:346-358   DOI
11 Garg AK, Kim JK, Owens TG, Ranwala AP, Choi YD, Kochian LV, Wu RJ (2002) Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses. Proc Natl Acad Sci USA 99:15898-15903   DOI
12 Goddijn OJM, van Dun K (1999) Trehalose metabolism in plants. Trends Plant Sci 4:315-319   DOI
13 Hinchee MAW, Connor-Ward DV, Newell CA, McDonnell RE, Sato SJ, Gasser CS, Fischhoff DA, Re DB, Fraley RT, Horsch RB (1988) Production of transgenic soybean plants using Agrobacterium-mediated gene transfer. Bio/Technol 6:915-922   DOI
14 Holmstrom KO, Mantyla E, Welin B, Mandal A, Palva ET, Tunnela OE, Londesborough J (1996) Drought tolerance in tobacco. Nature 379:683-684   DOI
15 Jang IC, Oh SJ, Seo JS, Choi WB, Song SI, Kim CH, Kim YS, Seo HS, Choi YD, Nahm BH, Kim JK (2003) Expression of a bifunctional fusion of the Escherichia coli genes for trehalose -6-phosphate synthase and trehalose-6-phosphate phosphatase in transgenic rice plants increases trehalose accumulation and abiotic stress tolerance without stunting growth. Plant Physiol 131:516-524   DOI
16 Jun SS, Yang JY, Choi HJ, Kim NR, Park MC, Hong YN (2005) Altered physiology in trehalose-producing transgenic tobacco plants : Enhanced tolerance to drought and salinity stresses. J Plant Biol 48:456-466   DOI
17 An G (1987) Binary Ti vectors for plant transformation and promoter analysis. Meth Enzymol 153:292-305   DOI
18 Chee PP (1990) Transformation of Cucumis sativus tissue by Agrobacterium tumefaciens and the regeneration of transformed plants. Plant Cell Rep 9:245-248
19 Cho MA, Song YM, Park YO, Ko SM, Min SR, Liu JR, Choi PS (2005a) The use of glufosinate as a selective marker for the transformation of cucumber (Cucumis sativus L.). Kor J Plant Biotechnol 32:161-165   과학기술학회마을   DOI
20 Cho MA, Song YM, Park YO, Ko SM, Min SR, Liu JR, Lee JH, Choi PS (2005b) Production of transgenic melon from the cultures of cotyledonary-node explant using Agrobacteriummediated transformation. Korean J Plant Biotech 32:257-262   과학기술학회마을   DOI
21 Dong JZ, Yang MZ, Jia SR, Chua NH (1991) Transformation of melon (Cucumis melo L.) and expression from the cauliflower mosaic virus 35S promoter in transgenic melon plants. Biotechnol 9:858-863   DOI
22 Drennan PM, Smith MT, Goldsworthy D, van Staden J (1993) The occurrence of trehalose in the leaves of the desiccation-tolerant angiosperm Myrothamnus flabellifolius. J Plant Physiol 142:493-496   DOI   ScienceOn
23 Elbein A (1974) The metabolism of ${\alpha},{\alpha}-trehalose$. Adv Carbohydr Chem Biochem 30:227-256.   DOI