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. 2018 Jul 10;9:920. doi: 10.3389/fpls.2018.00920

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Copyright © 2018 Majumder, Datta, Sarkar, Saha and Datta.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Qualitative analysis specific to chi11 gene and its functionality. (A) Gel diffusion assay- 150 μg crude protein pipetted into individual wells in the 1.5% agarose with 0.4% colloidal chitin gel plate. After incubation, lytic zones visualized by UV transillumination. #Not applicable for investigation. (B) A significant difference (^∗∗∗P = < 0.001) in mean zone area (calculated from three replicates) between transgenic and WT plants observed. (C) Fold change of developed zone area in transgenic plants over WT represents a significant difference (^∗∗∗P = < 0.001). (D) In-solution chitinase assay; 100 μg of total protein incubated with colloidal chitin and the resulting product measured at 530 nm. A significant difference (^∗∗∗P = < 0.001) in OD values between transgenic and non-transgenic WT plant observed. One-way ANOVA was used to compare the means and differences between group means at P < 0.05 and Tukey’s multiple comparisons taken under consideration by using Graphpad Prism 6 software. Here specific individual plants are the representatives of the three transgenic lines JBC1, JBC2, and JBC3. A total of 10 biological replicates were analyzed from each transgenic line.