Standardization of Agrobacterium tumefaciens-mediated genetic transformation protocol in Punica granatum L. cv. Kandhari Kabuli

Crop Research ◽  
2017 ◽  
Vol 52 (6) ◽  
pp. 268
Author(s):  
Nehanjali Parmar ◽  
Ajay Kumar Thakur ◽  
Kamlesh Kanwar
1970 ◽  
Vol 7 (2) ◽  
pp. 235-240
Author(s):  
MR Hossain ◽  
L Hassan ◽  
AK Patwary ◽  
MI Ferdous

A rapid, efficient and reproducible genetic transformation protocol was optimized for four aromatic rice varieties by using the established plant regeneration protocol. Mature embryos were inoculated with Agrobacterium tumefaciens strain EHA105 carrying a binary vector pIG121-Hm with GUS (reporter gene) and hpt (hygromycin resistance) gene and the transformation experiment was performed by optimizing two important parameters viz. infection times and co-cultivation periods. The highest response to GUS assay was showed by Kalizira (70% GUS positive) followed by Pusa Basmati 1 (66.67%) when the explants were inoculated for 25 minutes and co-cultivated for three days. Twenty five minutes infection time (9.44%) and three day co- cultivation period (8.06%) were found effective for percentage of transgenic shoot regeneration. The highest percentage of putative transgenic shoots was regenerated by variety Kalizira (33.33%) followed by Pusa Basmati 1 (20.0%) and Radhunipagol (13.33%). Among the varieties, Kalizira produced the highest percentage (60%) of rooted shoots. Kalizira also showed the highest survival rate in growth chamber (75%) and in field condition (60%). The performance of Tulsimala was poor for almost all the cases. Keywords: Agrobacterium; GUS; hpt gene; Transformation and Aromatic rice DOI: 10.3329/jbau.v7i2.4724 J. Bangladesh Agril. Univ. 7(2): 235-240, 2009


2020 ◽  
Author(s):  
Araceli Oropeza-Aburto ◽  
Sergio Alan Cervantes-Perez ◽  
Victor A Albert ◽  
Luis Rafael Herrera-Estrella

Abstract Background The genus Utricularia belongs to Lentibulariaceae, the largest family of carnivorous plants, which includes terrestrial, epiphytic and aquatic species. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologists since Darwin‘s early studies. Utricularia gibba is itself an aquatic plant with sophisticated bladder traps having one of the most complex suction mechanisms for trapping prey. However, the molecular characterization of the mechanisms that regulate trap development and the biophysical processes involved in prey trapping are still largely unknown due to the lack of a simple and reproducible gene transfer system. Results Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens . An in vitro selection system using Phosphinotricin as a selective agent was established for U. gibba . Plant transformation was confirmed by histochemical GUS assays and PCR and qRT-PCR analyses. We report on the expression pattern of the 35S promoter and of the promoter of a trap-specific ribonuclease gene in transgenic U. gibba plants. Conclusions The genetic transformation protocol reported here is an effective method for studying developmental biology and functional genomics of this genus of carnivorous plants and advances the utility of U. gibba as a model system to study developmental processes involved in trap formation.


2020 ◽  
Author(s):  
Araceli Oropeza-Aburto ◽  
Sergio Alan Cervantes-Perez ◽  
Victor A Albert ◽  
Luis Rafael Herrera-Estrella

Abstract Background The genus Utricularia belongs to Lentibulariaceae, the largest family of carnivorous plants, which includes terrestrial, epiphytic and aquatic species. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologists since Darwin‘s early studies. Utricularia gibba is itself an aquatic plant with sophisticated bladder traps having one of the most complex suction mechanisms for trapping prey. However, the molecular characterization of the mechanisms that regulate trap development and the biophysical processes involved in prey trapping are still largely unknown due to the lack of a simple and reproducible gene transfer system. Results Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens . An in vitro selection system using Phosphinotricin as a selective agent was established for U. gibba . Plant transformation was confirmed by histochemical GUS assays and PCR and qRT-PCR analyses. We report on the expression pattern of the 35S promoter and of the promoter of a trap-specific ribonuclease gene in transgenic U. gibba plants. Conclusions The genetic transformation protocol reported here is an effective method for studying developmental biology and functional genomics of this genus of carnivorous plants and advances the utility of U. gibba as a model system to study developmental processes involved in trap formation.


2019 ◽  
Vol 46 (3) ◽  
pp. 223-230
Author(s):  
Atmitri Sisharmini ◽  
Bambang Sapta Purwoko ◽  
Nurul Khumaida ◽  
Dan Kurniawan Rudi Trijatmiko

Protocols for genetic transformation of rice have been widely developed, however the protocols are not universal and inapplicable for all types of rice plants directly. Transformation protocol on rice cv. Fatmawati needs to be developed to generate transgenic lines. The present research was carried out to optimize genetic transformation protocol in rice cv. Fatmawati mediated by Agrobacterium tumefaciens harboring pCambia1301 construct using immature embryo as an explant. The experiment was arranged in a completely randomized design. Factors influencing efficiency of transformation, i.e., sensitivity of callus to hygromycin antibiotic, acetosyringone concentration used in cultivation medium, hygromycin concentration for transformant selection were optimized. The results showed that genetic transformation of rice cv. Fatmawati mediated by A. tumefaciens using immature embryos have been successfully carried out with several parameters. Addition of 100 µM acetosyringone in co-cultivation medium and 30 mg L-1 hygromycin for transformant callus selection were optimal for genetic transformation of rice cv. Fatmawati mediated by A. tumefaciens. Transformation efficiency was found to be 7.84% based on the lines carrying the hpt gene. This result would be a valuable reference in genetic transformation of rice cv. Fatmawati using target genes.Keywords: immature embryo, Oryza sativa, pCambia1301, transformation efficiency


2020 ◽  
Author(s):  
Araceli Oropeza-Aburto ◽  
Sergio Alan Cervantes-Perez ◽  
Luis Rafael Herrera-Estrella

Abstract Background Utricularia genus belongs to Lentibulariaceae family and has the highest number of species including terrestrials, epiphytes and aquatics plants. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologist since the early studies of Darwin. Utricularia gibba is an aquatic carnivorous plant with sophisticated bladders traps that have one of the most complex suction mechanisms for trapping a prey. However, the molecular characterization of trap developmental and the biophysical processes involved in prey trapping are still largely unknown due to the lack of simple and reproducible gene transfer system for carnivorous plants.Results Here, we report the establishment of a simple, fast and effective protocol for the genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens . An in vitro selection system using Phosphinotricin as selective agent was established for U. gibba. We report the tissue specific expression of the 35S promoter and the promoter of a trap specific ribonuclease gene. Plant transformation was confirmed by PCR and Real Time PCR in U. gibba plants.Conclusions We conclude that the genetic transformation protocol we developed is an effective method to study developmental biology and functional genomics of carnivory and propose U. gibba as model to study the developmental processes involved in trap formation.


2000 ◽  
Vol 108 (4) ◽  
pp. 413-419
Author(s):  
Patricia Dupré ◽  
Jerôme Lacoux ◽  
Godfrey Neutelings ◽  
Dominique Mattar-Laurain ◽  
Marc-André Fliniaux ◽  
...  

2019 ◽  
pp. 7-14
Author(s):  
Hai Trieu Ly ◽  
Tuan Anh Vo ◽  
Viet Hong Phong Nguyen ◽  
Thi My Sa Pham ◽  
Bich Thao Lam ◽  
...  

Background: The natural antioxidants have an important role in the prevention of many diseases. The aim of study is to investigate phytochemical components, antioxidant activity and acute oral toxicity of Pomegranate (Punica granatum L.) fruit peel (PFP) extract. Materials and methods: Phytochemicals of PFP were determined by qualitative chemical tests, thin layer chromatography, total polyphenol and flavonoid contents. The PFP extract was evaluated for antioxidant activity by DPPH assay and MDA assay. In vivo acute oral toxicity test was conducted using Karber-Behrens method to determine LD50. Results: Results illustrated that PFP mainly contains flavonoids, alkaloids, tannins, triterpenes, saponins, and coumarins. PFP extract exhibited the total polyphenol and flavonoid contents with 189.97 mg gallic acid equivalent/g dry weight and 9.42 mg quercetin equivalent/g dry weight, respectively. The DPPH free radical scavenging and anti-lipid peroxidation activities of PFP extract were expressed with IC50 value of 4.80 μg/mL and 0.38 μg/ mL, sequentially. Simultaneously, the Dmax (the maximum dose administered to mice that no toxicity was observed) of PFP extract was determined to be 21.28 g/kg, equivalent to 35.64 g dried herb. Conclusion: The PFP extract is relatively safe and revealed high antioxidant activity. Key words: Punica granatum L.; polyphenols; flavonoids; gallic acid; quercetin; antioxidant activity; acute oral toxicity


2016 ◽  
Vol 45 (11) ◽  
pp. 1533-1543
Author(s):  
Ju-Ryun Na ◽  
Sunoh Kim ◽  
Ara Jo ◽  
Donghyuck Bae ◽  
Kyo-Nyeo Oh ◽  
...  

2021 ◽  
Vol 352 ◽  
pp. 129396
Author(s):  
Clarisse Gosset-Erard ◽  
Minjie Zhao ◽  
Sonia Lordel-Madeleine ◽  
Saïd Ennahar

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