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        Agrobacterium-Mediated Cry1A(b) Gene Transfer in Punica granatum L. cv. Kandhari Kabuli Using Different In Vitro Regeneration Pathways

        Verma, Vipasha,Kanwar, Kamlesh,Tufchi, Mahak,Kashyap, Monika 한국작물학회 2014 Journal of crop science and biotechnology Vol.17 No.1

        Three different regeneration systems, viz. regeneration through callus cultures using embryonic explant, direct regeneration using shoot bud/nodal segments as explant and regeneration through cell suspension culture using cotyledonary explant (for the induction of transgenic callus for suspension culture) were evaluated to see their effect on transfer of Cry1A(b) gene to Punica granatum L. cv. Kandhari Kabuli through Agrobacterium mediated transformation. Pre-conditioning and co-cultivation durations had a marked effect on transformation frequency of different explants. Out of different explants used (embryo, shoot bud, and cotyledon) for different regeneration systems cotyledonary explant showed highest putative transformation frequency (13.54%) inducing callus on selective medium for carrying out cell suspension culture to regenerate transgenic shoots. Despite of the highest transformation frequency obtained from the cotyledon explant, the plating efficiency of the transgenic cells generated through the transgenic callus (callus formed from the cotyledonary explant) during cell suspension culture was found to be very low (0.7%). Thus the plating efficiency has also played worth mentioning role in the regeneration of transformants following cell suspension culture. Among the three regeneration systems, regeneration through callus cultures using embryonic explant was found to be best for regeneration of transformants. The highest per cent regeneration of 23.33 was obtained from the putative transgenic embrogenic calli. Successful genetic transformation in the transformed plantlets was confirmed by PCR analysis. The transformation system thus developed is valuable and may be used to produce insect resistant trees.

      • KCI등재

        Agrobacterium-Mediated Cry1A(b) Gene Transfer in Punica granatum L. cv. Kandhari Kabuli Using Different In Vitro Regeneration Pathways

        Vipasha Verma,Kamlesh Kanwar,Mahak Tufchi,Monika Kashyap 한국작물학회 2014 Journal of crop science and biotechnology Vol.17 No.1

        Three different regeneration systems, viz. regeneration through callus cultures using embryonic explant, direct regeneration usingshoot bud/nodal segments as explant and regeneration through cell suspension culture using cotyledonary explant (for the inductionof transgenic callus for suspension culture) were evaluated to see their effect on transfer of Cry1A(b) gene to Punica granatum L. cv. Kandhari Kabuli through Agrobacterium mediated transformation. Pre-conditioning and co-cultivation durations had a marked effecton transformation frequency of different explants. Out of different explants used (embryo, shoot bud, and cotyledon) for differentregeneration systems cotyledonary explant showed highest putative transformation frequency (13.54%) inducing callus on selectivemedium for carrying out cell suspension culture to regenerate transgenic shoots. Despite of the highest transformation frequencyobtained from the cotyledon explant, the plating efficiency of the transgenic cells generated through the transgenic callus (callusformed from the cotyledonary explant) during cell suspension culture was found to be very low (0.7%). Thus the plating efficiencyhas also played worth mentioning role in the regeneration of transformants following cell suspension culture. Among the three regenerationsystems, regeneration through callus cultures using embryonic explant was found to be best for regeneration of transformants. The highest per cent regeneration of 23.33 was obtained from the putative transgenic embrogenic calli. Successful genetic transformationin the transformed plantlets was confirmed by PCR analysis. The transformation system thus developed is valuable and maybe used to produce insect resistant trees

      • KCI등재

        Interferon-gamma release assay has poor diagnostic accuracy in differentiating intestinal tuberculosis from Crohn’s disease in tuberculosis endemic areas

        Karan Sachdeva,Peeyush Kumar,Bhaskar Kante,Sudheer K. Vuyyuru,Srikant Mohta,Mukesh K. Ranjan,Mukesh K. Singh,Mahak Verma,Govind Makharia,Saurabh Kedia,Vineet Ahuja 대한장연구학회 2023 Intestinal Research Vol.21 No.2

        Background/Aims: Intestinal tuberculosis (ITB) and Crohn’s disease (CD) frequently present with a diagnostic dilemma because of similar presentation. Interferon-gamma release assay (IGRA) has been used in differentiating ITB from CD, but with sparse reports on its diagnostic accuracy in tuberculosis endemic regions and this study evaluated the same. Methods: Patients with definitive diagnosis of ITB (n=59) or CD (n=49) who underwent IGRA testing (n=307) were retrospectively included at All India Institute of Medical Sciences, New Delhi (July 2014 to September 2021). CD or ITB was diagnosed as per standard criteria. IGRA was considered positive at >0.35 IU/mL. Relevant data was collected and IGRA results were compared between ITB and CD to determine its accuracy. Results: Among 59 ITB patients (mean age, 32.6±13.1 years; median disease duration, 1 year; male, 59.3%), 24 were positive and 35 tested negative for IGRA. Among 49 CD patients (mean age, 37.8±14.0; median disease duration, 4 years; male, 61.2%), 12 were positive and 37 tested negative for IGRA. Hence, for diagnosing ITB, IGRA showed a sensitivity, specificity, positive and negative predictive values of 40.68%, 75.51%, 66.67%, and 51.39%, respectively. The area under the curve of IGRA for ITB diagnosis was 0.66 (95% confidence interval, 0.55–0.75). In a subset (n=64), tuberculin skin test (TST) showed sensitivity, specificity, positive and negative predictive values of 64.7%, 73.3%, 73.3%, and 64.71%, respectively. IGRA and TST were concordant in 38 (59.4%) patients with κ=0.17. Conclusions: In a tuberculosis endemic region, IGRA had poor diagnostic accuracy for differentiating ITB from CD, suggesting a limited value of IGRA in this setting.

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