Publications of NIPGR Scientists

Permanent URI for this communityhttps://ndkr-library.nipgr.ac.in/handle/123456789/1

Browse

Subject: search.filters.subject.GUS expression

Search Results

Now showing 1 - 2 of 2
  • Thumbnail Image
    Item
    Two homeologous MATE transporter genes, NtMATE21 and NtMATE22, are involved in the modulation of plant growth and flavonol transport in Nicotiana tabacum
    (Oxford University Press, 2022) Gani, Umar; Nautiyal, Abhishek Kumar; Kundan, Maridul; Rout, Biswaranjan; Pandey, Ashutosh; Misra, Prashant
    Multidrug and toxic compound extrusion (MATE) family has been implicated in the transport of a diverse range of molecules, including specialized metabolites. In Nicotiana tabacum, only a limited number of MATE transporters have been functionally characterized, and no MATE transporter has been studied in the context of flavonoid transport in this plant species so far. Here in the present study, we characterize two homeologous MATE genes, NtMATE21 and NtMATE22, and demonstrate their role in flavonol transport and in plant growth and development. The expression of these two genes was reported to be up-regulated in trichomes as compared to the trichome-free leaf. The transcript levels of NtMATE21 and NtMATE22 were found to be higher in flavonol over-producing tobacco transgenic lines as compared to wild type (WT) tobacco. The two transporters were demonstrated to be localized to the plasma membrane. Genetic manipulation of NtMATE21 and NtMATE22 led to altered growth phenotypes and modulated flavonol contents in N. tabacum. The GUS and GFP fusion transgenic lines of promoter regions suggested that NtMATE21 and NtMATE22 exclusively express in the trichome heads in the leaf tissue and petals. Moreover, in transient transactivation assay, NtMYB12, a flavonol-specific MYB transcription factor, was found to transactivate the expression of NtMATE21 and NtMATE22 genes. Together, our results strongly suggest the involvement of NtMATE21 and NtMATE22 in flavonol transport as well as in the regulation of plant growth and development.
  • Thumbnail Image
    Item
    Genotype independent regeneration and agrobacterium-mediated genetic transformation of sweet potato (Ipomoea batatas L.)
    (Bangladesh Association for Plant Tissue Culture and Biotechnology, 2013) Shekhar, Shubhendu; Agrawal, Lalit; Buragohain, Alak Kumar; Datta, Asis; Chakraborty, Subhra; Chakraborty, Niranjan
    Development of an efficient genotype independent regeneration and genetic transformation system in sweet potato continues to be of great interest. Agrobacterium‐mediated genetic transformation protocol was established in two different cultivars of sweet potato using Agrobacterium strain EHA105 harbouring binary plasmid pBI121 containing GUS and nptII genes. The internodal stem segments from 30‐day‐old micropropogated plants were used as explant with different combinations of media and hormones. MS and LS media with various concentrations of growth regulators proved to be non‐responsive and the infecundity was severe with the addition of cytokinins. Nonetheless, MS with 2,4‐D and TDZ gave a good percentage of callusing but with low differentiation. In different concentrations of NAA, significant amount of callusing was observed but percentage of rooting remained low in both the genotypes. Gamborg’s B5 supplemented with NAA proved to be the most suitable media and hormone combination, which yielded shoot formation after 8 ‐ 10 weeks with a regenera‐ tion efficiency of 40 ‐ 70%. Stable integration of transgene was confirmed by PCR analysis. Furthermore, qRT‐PCR analysis was performed to assess the transcript accumulation in addition to the GUS enzymatic assay in the transgenic lines.