International Journal of Horticulture, 2017, Vol.7, No. 6, 40-46
45
conducted by
(Lipsky et al., 2014) where genetic transformation of
Ornithogalum
via particle bombardment was
carried out. In our study all leaf sections were dipped in
gus
buffer and left the leaves in assay solution at 37˚C for
48 hrs. Temperature is also an important factor for highest activity of GUS assay (Su et al., 2012). After 48 hrs,
Gus expressing cells showed blue colour as a result of chromogenic cleavage by X-Gluc, representing successful
expression of gus gene (Figure 3). The Gus assay is safe, highly reliable, versatile, and easy to perform and needs
no specialized equipment (Jefferson et al., 1987; Jefferson and Wilson, 1991).
References
Becker D., Brettschneider R., and Loerz H., 1994, Fertile transgenic wheat from microprojectile bombardment of scutellar tissue, The Plant Journal, 5: 299-307
PMid:8148881
Brozynska M., Furtado A., and Henry R.J., 2015, Genomics of crop wild relatives: expanding the gene pool for crop improvement, Plant Biotechnology Journal,
1–16
Conner A.J., and Meredith C.P., 1989, Genetic manipulation of plant cells, In: Marcus A, Ed. The Biochemistry of plants, Vol. 15: Molecular biology, Pp.
653-688 Academic Press
Davey M.R., Lowe K.C., Power J.B., and Anthony P., 2000, Protoplast fusion for the generation of unique plants, Encyclopedia of cell technology
Filipecki M., and Malepszy S., 2006, Unintended consequences of plant transformation, Journal of Molecular and Applied Genetics, 47: 277-286
PMid:17132892
Fior S., and Gerola P.D., 2009, Impact of ubiquitous inhibitors on the GUS gene reporter system: evidence from the model plants Arabidopsis, tobacco and rice
and correction methods for quantitative assays of transgenic and endogenous GUS, Plant Methods 5:19
PMid:20042078 PMCid:PMC2805642
Ford-Lloyd B.V., Schmidt M., Armstrong S.J., Barazani O., Engels J., Hadas R., ... and Li Y., 2011, Crop wild relatives—undervalued, underutilized and under
threat?, BioScience, 61(7), 559-565
Janssen B.J., and Gardner R.C., 1990, Localized transient expression of GUS in leaf discs following cocultivation with Agrobacterium, Plant Molecular
Biology, 14: 61-72
PMid:2101312
Jefferson R.A., Kavanagh T.A., and Bevan M.W., 1987, GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants,
EMBO Journal, 6: 3901–3907
PMid:3327686 PMCid:PMC553867
Jefferson R.A., and Wilson K.J., 1991, The GUS gene fusion system, In S.B. Gelvin and R.A. Schilperoort (eds.) Plant Molecular Biology Manual; B14: Pp.
1-33 Kluwer Academic Publishers (Dordrecht)
Joyia F.A., and Khan M.S., 2013, Scutellum-derived callus-based efficient and reproducible regeneration system for elite varieties of indica rice in Pakistan,
International Journal of Agriculture and Biology, 15: 27-33
Kettenring K.M., Mercer K.L., Reinhardt A.C., and Hines J., 2014, Editor’s Choice: Application of genetic diversity–ecosystem function research to ecological
restoration, Journal of Applied Ecology, 51: 339-348
Klein T., Gradziel T., Fromm M., and Sanford J., 1988, Factors Influencing Gene Delivery into Zea Mays Cells by High–Velocity Microprojectiles, Nature
Biotechnology, 6: 559-563
Li B., Huang W., and Bass T., 2003, Shoot production per responsive leaf explant increases exponentially with explant organogenic potential in Nicotiana
species, Plant Cell Reports, 22: 231–238
PMid:12920564
Lipsky A., Cohen A., Ion A., and Yedidia I., 2014, Genetic transformation of
Ornithogalum
via particle bombardment and generation of
Pectobacterium
carotovorum
-resistant plants, Plant Science, 228: 150-158
PMid:25438795
Maliga P., 1982, Cell culture procedures for
Nicotiana plumbaginifolia,
Plant Molecular Biology Newsletters, 3:88-94
Murashige T., and Skoog F., 1962, A revised medium for rapid growth and bioassays with tobacco tissue culture, Physiologia Plantarum, 15: 473–497
Nikova V.M., Zagorska N., and Pundeva R., 1991, Development of four tobacco cytoplasmic male sterile sources using
in vitro
techniques, Plant Cell Tissue
and Organ Culture, 27(3): 289-295
