To stain cellulose, Mitra and Loque [61] protocol for Calcofluor staining was followed with some modifications. Briefly, the roots were transferred to 2 ml tubes and stained with 0.02% calcoflour white (Sigma) for 5 min. The primary root was placed on the microscope glass slide and transverse section of middle part of the roots was prepared using sharp blade. Calcofluor White was visualized using an epifluorescence microscope (Zeiss Imager M2).
The staining of callose was done according to Muller et al. [62] with some modifications. Briefly, the roots were stained with 0.1% (w/v) aniline blue solution in 0.1 M sodium phosphate buffer (pH = 7.2) for 1.5 h. The cross-section of roots was done as described above and visualized under epifluorescence microscope (Zeiss Imager M2).
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