Agrobacterium tumefaciens-mediated genetic transformation of a recalcitrant grain legume, lentil (Lens culinaris Medik)

Date

2009-03-01

Author

Akcay, Ufuk Celikkol
Mahmoudian, M.
Kamci, H.
YÜCEL, MUSTAFA
Öktem, Hüseyin Avni

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

163
views

0
downloads

A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient beta-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T-0, T-1, T-2 and T-3 plants. PCR assays of T-1, T-2 and T-3 progenies confirmed the stable transmission of the transgene to the next generations.

Subject Keywords

Agronomy and Crop Science, Plant Science, General Medicine

URI

https://hdl.handle.net/11511/46705

Journal

PLANT CELL REPORTS

DOI

https://doi.org/10.1007/s00299-008-0652-4

Collections

Department of Biology, Article

Suggestions

OpenMETU
Core

Micromeria myrtifolia: The influence of the extracting solvents on phenolic composition and biological activity Sarikurkcu, Cengiz; Hanine, Hafida; Sarikurkcu, Rabia Busra; Sarikurkcu, Rifat Tayyib; Amarowicz, Ryszard (Elsevier BV, 2020-03-01) Currently, there is much interest in using the active compounds that exist in plants as natural treatments. The pharmaceutical industry is also looking for novel chemical compounds from natural sources that do not have side effects. The purpose of this study was to identify the antioxidant and enzymatic inhibition activities of different extracts (ethyl acetate, methanol and water) of Micromeria myrtifolia from Turkey. The phenolic constituents in the extracts were also identified using liquid chromatograph...
Bioactivity-guided isolation of cytotoxic secondary metabolites from the roots of Glycyrrhiza glabra and elucidation of their mechanisms of action Cevik, Dicle; Yilmazgoz, S. Burcin; KAN, YÜKSEL; Guzelcan, Ece Akhan; Durmaz, Irem; Atalay, Rengül; KIRMIZIBEKMEZ, HASAN (Elsevier BV, 2018-11-15) Licorice (Glycyrrhiza glabra L.) is one of the most widely used plants worldwide for its various pharmacological activities. The aim of this study was to isolate the potential cytotoxic secondary metabolites from the MeOH extract prepared from the roots of Glycyrrhiza glabra through bioactivity-guided isolation procedure and to elucidate their mechanisms of action. The crude MeOH extract as well as CHCl3 and EtOAc subextracts significantly inhibited cell proliferation on hepatocelullar (Huh7), breast (MCF7)...
Ionic liquid pretreatment allows utilization of high substrate loadings in enzymatic hydrolysis of biomass to produce ethanol from cotton stalks Haykir, Nazife Isik; Bakir, Ufuk (Elsevier BV, 2013-11-01) To enhance the enzymatic conversion of cotton stalks to glucose at high substrate loadings and subsequently to produce ethanol, biomass was subjected to ionic liquid and alkaline pretreatments. Ionic liquid, 1-ethyl-3-methylimidazolium acetate (EMIMAc), which reduced biomass crystallinity, was effective in improving the conversion of cotton stalks to glucose (82-85%) regardless of the substrate loading used during enzymatic hydrolysis. On the other hand, the glucose yield decreased from 67% to 55% with an i...
Detection of physically interacting proteins with the CC and NB-ARC domains of a putative yellow rust resistance protein, Yr10, in wheat Yildirim-Ersoy, Figen; Ridout, Christopher J.; Akkaya, Mahinur (Springer Science and Business Media LLC, 2011-08-01) Gene-for-gene (GFG) resistance is a potent defense mechanism in plants, that is mediated by resistance (R) proteins. In GFG resistance, pathogen effector or avirulence (Avr) proteins are recognised by R-proteins which initiate a series of signal transduction events that lead to hypersensitive cell death. In cereals, many R-proteins are comprised of an N-terminal coiled-coiled (CC) domain, a Nucleotide Binding (NB) domain and a Leucine Rich Repeats (LRR) region associated with effector recognition. NB-LRR im...
Experimental study of solute transport and extraction by a single root in soil Kim, J; Chang, YS; Corapcioglu, MY; Drew, MC (Springer Science and Business Media LLC, 2005-02-01) The fate of C-14-2,4,6-trinitrotoluene ([U-C-14]TNT) in soil/plant systems was studied using onion (Allium cepa L.) plants with only a single root. It was found that the single roots grew exponentially and that the rate of water uptake of the onion plants increased exponentially, as well. The concentration of [U-C-14] in the roots at first increased and then appeared to reach a steady state, while the [U-C-14] concentration in the leaves was found to increase linearly with time. The [U-C-14] concentration i...

Citation Formats

U. C. Akcay, M. Mahmoudian, H. Kamci, M. YÜCEL, and H. A. Öktem, “Agrobacterium tumefaciens-mediated genetic transformation of a recalcitrant grain legume, lentil (Lens culinaris Medik),” PLANT CELL REPORTS, pp. 407–417, 2009, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/46705.