Determination of Cry1Ac copy number in transgenic pigeonpeaplants using quantitative real time PCR.

Jain, M.; Khatodia, Surender; Kharb, Pushpa; Batra, Parveen; Chowdhury, V. K.

doi:10.18805/lr.v0iof.11300

Cited by 4 publications

(10 citation statements)

References 15 publications

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“…4A), Lanes 1-5). Similar results for Southern hybridization have been shown for Vigna mungo (Bhomkar et al, 2008), Cicer arietinum (Ghanti et al, 2011;Khatodia et al, 2014), Vicia faba (Hanafy et al, 2013) and Cajanus cajan (Jain et al, 2017) revealed stable integration of transgene into transgenic plant genome whereas no signal was recorded in non-transformed control (WT) plants.…”

Section: Southern Hybridization Analysissupporting

confidence: 78%

“…It was found that real-time PCR analysis was significantly correlated with Southern blot analysis and both showed single copy insertion in respective transgenic chickpea plants. Similarly, single copy insertion was observed in transgenic chickpea (Khatodia et al, 2014) and pigeonpea (Jain et al, 2017).…”

Section: Copy Number Comparison Studiesmentioning

confidence: 75%

“…2b). PCR based screening of putative transgenics has been widely used by various researchers to confirm the incorporation of the transgene in legumes like cowpea (Mishra et al, 2014), peanut (Singh et al, 2014), pigeonpea (Jain et al, 2017) and chickpea (Mubina et al, 2018).…”

Section: Screening Of Putative Transgenicsmentioning

confidence: 99%

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Engineering Chickpea Variety HC-1 with OsRuvB Gene for Salt Stress Tolerance

Preeti¹,

Kharb²

2020

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Background: Salinity is a major problem worldwide and is increasing day by day. Salt stress causes severe yield losses in crop plants and the damages in chickpea are up to 100%. To overcome these losses, the present study was undertaken to develop transgenic chickpea plants (var. HC-1) carrying OsRuvB gene for salt stress tolerance. Methods: Transgenic chickpea plants harboring OsRuvB gene were developed using Agrobacterium-mediated transformation. T0 putative transgenic chickpea plants were screened for the presence of OsRuvB gene through PCR using gene specific primers. The stable integration and copy number of transgene in transgenic chickpea plants were confirmed through Southern hybridization and qRT-PCR. T1 generation transgenic chickpea plants were screened for the presence of OsRuvB gene using direct PCR (Phire Direct PCR kit). Result: PCR-based screening of putative transformants using gene-specific primers showed a transformation frequency of 17%. Southern blot and real-time PCR analysis revealed stable and single-copy insertion. In T1 generation a total of 74 plants (out of 170) showed the presence of OsRuvB gene. The engineered lines developed in the present investigation can be further undertaken to develop transgenic chickpea plants for salt stress tolerance.

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Section: Southern Hybridization Analysissupporting

confidence: 78%

Section: Copy Number Comparison Studiesmentioning

confidence: 75%

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Engineering Chickpea Variety HC-1 with OsRuvB Gene for Salt Stress Tolerance

Preeti¹,

Kharb²

2020

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“…For developing other transgenic lines mentioned in Table 2 [ 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ], the seeds were directly transferred to potted soil after co-cultivation, omitting the step of germinating in the presence of cefotaxime. For wheat and rice transformation, 200 µM acetosyringone was added to the bacterial suspension during co-cultivation.…”

Section: Resultsmentioning

confidence: 99%

A Genotype-Independent, Simple, Effective and Efficient in Planta Agrobacterium-Mediated Genetic Transformation Protocol

Kharb

Chaudhary

Tuteja

et al. 2022

MPs

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Crop improvement under changing climatic conditions is required to feed the growing global population. The development of transgenic crops is an attractive and conceivably the most effective approach for crop improvement with desired traits in varying climatic situations. Here, we describe a simple, efficient and robust in planta Agrobacterium-mediated genetic transformation method that can be used in most crops, including rice, wheat and cotton, and particularly in tissue culture recalcitrant crops, such as chickpea and pigeon pea. The protocol was successfully used for the development of transgenic chickpea and pigeon pea lines for resistance against pod borer. Transgenic lines in chickpea, pigeon pea and wheat were also developed for salt stress tolerance. These lines exhibited improved salt tolerance in terms of various physio-biochemical parameters studied. Since the protocol is rapid, as no tissue culture step is involved, it will significantly contribute to the improvement of most crops and will be of interest for plant biologists working with genetic engineering or genome editing.

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“…The percentage of GC content of DNA has a significant effect on its Tm. Because the higher the percentage of GC content, the higher its Tm Jain et al (2017).…”

Section: Methodsmentioning

confidence: 99%

Analysis of the effects of coumarin on Lens culinaris Medik by some biochemical parameters using real-time polymerase chain reaction

Yüksel

Aksoy

2019

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This study aimed to reveal the effects of coumarin on Lens culinaris Medik. The germinated and ungerminated seed counts of the experimental groups were determined and the EC50 value was calculated by probit analysis. In biochemical studies, catalase and superoxide dismutase enzyme activities were investigated together with lipid peroxidation and hydrogen peroxide quantities. The results demonstrated an increase in the amount of malondialdehyde, a measure of lipid peroxidation, a decrease in the amount of H2O2 and EC50 value in the CAT activity and an increase in the EC50 x 2 value. In real-time PCR analysis, three different genetic expressions related to abiotic stress (CAT, Cu / Zn SOD and Mn SOD gene expression) were examined. It was determined that coumarin caused genotoxic and biochemical damage on L. culinaris.

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Determination of Cry1Ac copy number in transgenic pigeonpeaplants using quantitative real time PCR.

Cited by 4 publications

References 15 publications

Engineering Chickpea Variety HC-1 with OsRuvB Gene for Salt Stress Tolerance

Engineering Chickpea Variety HC-1 with OsRuvB Gene for Salt Stress Tolerance

A Genotype-Independent, Simple, Effective and Efficient in Planta Agrobacterium-Mediated Genetic Transformation Protocol

Analysis of the effects of coumarin on Lens culinaris Medik by some biochemical parameters using real-time polymerase chain reaction

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