Eos 760d camera

Side-view photographs of all plants were taken manually after 7, 14, 21, and 28 days post-transplanting (dpt) using an EOS 760D camera coupled with an EF-S 17–85 mm lens (Canon, Tokyo, Japan). Plant height was measured using ImageJ software [71 (link)]. After 31 dpt, all leaves of > 2 cm in length were detached, and top-view images were taken as above. The total number of leaves were counted, and leaf area were calculated with Morpho Analysis software. At the same time, roots and shoots from each plant were separately harvested and immediately weighed to determine their fresh weight. Dry weight was measured after a 48-h incubation in an oven at 80 °C. The shoot/root ratio = (leaf dry mass + stem dry mass)/root dry mass, of each plant was calculated [29 (link), 72 (link)].

The optical and morphological properties of fibers are analyzed by polarizing optical microscopy (POM) and scanning electron microscopy (SEM). POM is done in transmission mode using an Olympus BX51 equipped with a Linkam (T95 series LTS120E, Surrey, UK) heating/cooling stage, an Olympus DP73 camera (Tokyo, Japan) and a Canon EOS760D camera (Tokyo, Japan). SEM imaging of the fibers are done using JEOL JSM-6010LA (Akishima, Japan) being operated in 12–15 kV range using an In-lens secondary electron detector. For SEM imaging fiber samples are gold coated (≈25 nm thickness) using a sputter coater (Balzers SCD 050 Sputter Coater) for 100 s. SEM image analysis is done using ImageJ^® software to calculate the fiber diameter [42 (link)]. The average fiber diameter is calculated by analyzing 50 fibers from different sample locations. Infrared spectroscopy (IR) is done using a Thermo Scientific^TM Nicolet^TM iS5 spectrometer with id5 ATR accessory.